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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareAWARENESS, ATTITUDE AND PRACTICE OF PHARMACOVIGILANCE AMONG HEALTHCARE
PROFESSIONALS AND STUDENTS IN A TERTIARY CARE TEACHING HOSPITAL
English0614Apurva AgrawalEnglish Parul ChaturvediEnglishAims: To find out the level of awareness and attitude towards pharmacovigilance and extent of ADR
reporting in healthcare professionals and medical students. Materials and method: A total of 799 participants including healthcare professionals and students were asked to fill a predesigned questionnaire. It consisted of questions regarding awareness, attitude and practice of pharmacovigilance. Data collected were analyzed using relevant statistical tests. Awareness between healthcare professionals and students was compared using chi square test. Results: 70.46% of participants responded to the questionnaire. 22% of doctors and 37% of nurses had reported ADR to any authority in last 2 years. Lack of awareness about the ADR reporting system was the most common reason for non-reporting. Majority of healthcare professionals and students considered ADR reporting as very important and recommended active involvement of pharmacovigilance in medical curriculum. Conclusion: Overall level of awareness was low both among healthcare professionals and students. There is a great need to increase the awareness and improve the attitude of healthcare professionals and students towards pharmacovigilance and its national programme. Regular training sessions and awareness campaigns need to be conducted. Pharmacovigilance should be included in the undergraduate training of MBBS, pharmacy, nursing and physiotherapy students.
EnglishADR reporting, healthcare professionals, medical students, PharmacovigilanceINTRODUCTION
Pharmacovigilance as described by WHO is detection, assessment, understanding and prevention of adverse effects or any other drug related problem.[1] Adverse drug reactions (ADRs) are associated with significant morbidity and mortality, and are an important cause of hospitalizations.[2] Studies have shown ADRs as fourth major cause of death in USA, but such data are lacking in India.[3] As health care professionals are the first one who are in contact with patients taking drugs, spontaneous reporting by them is an effective way to generate early signals of ADRs. It is the most practical way to detect rare adverse events, adverse events caused by prolonged use of drugs and many drug-drug interactions. [4] Thus, awareness among healthcare workers and their attitude towards pharmacovigilance are important determinants of ADR reporting rate. WHO has developed a system for reporting of ADRs by establishment of International Drug Monitoring Programme, coordinated by Uppsala Monitoring Centre, Sweden. In India also National Pharmacovigilance Programme (NPP) was started in 2004.[5] This programme was relaunched in 2010 as Pharmacovigilance Programme of India (PvPI), and is now coordinated by the Indian Pharmacopoeia Commission, Ghaziabad.[6] Still pharmacovigilance is in its infancy phase in India and under reporting is a major problem. Studies done in other countries also reveal under reporting of ADRs.[7-10] Lack of awareness among health care professionals is one of the reasons for under reporting. Thus, to improve the ADR reporting rate, it is important to improve the awareness, attitude and practices of the healthcare professionals regarding pharmacovigilance. The best time to do this is during undergraduate training of students of MBBS, pharmacy and nursing. This will help by developing a culture of ADR reporting in healthcare workers. Though studies reporting the level of awareness and practices of pharmacovigilance have been done in other countries, [7, 8, 11, 12] very few studies have focused this aspect in India, and during literature search no such study was found from Rajasthan. Further most of the studies have included healthcare workers, but studies on awareness among undergraduate students are limited. [9] Moreover, there was no peripheral centre of NPP in Rajasthan. Recently a new ADR Monitoring Center under PvPI has been established in a Medical College of Rajasthan. [6] Hence, the present study was conducted to develop a baseline data of awareness, attitude and practice of pharmacovigilance in health care professionals and medical students in a Tertiary Healthcare Teaching Hospital in Rajasthan.
MATERIAL AND METHODS
This was a cross sectional, observational, questionnaire based study conducted in a Tertiary Care Teaching Hospital of Rajasthan. Duration of study was of 2 months from 1st May 2011 to 30th June 2011. Total 799 participants were approached, which included health care professionals and medical students. All healthcare professionals working in the Hospital including clinicians, pharmacists and nursing staff were included. Medical students of MBBS, students of Nursing College, Pharmacy College and Physiotherapy College attached to the Hospital were included in the study. All those who denied participation in the study and students who have not been introduced to Pharmacology (e.g. students who were in 1st year of their course) were excluded from the study. Procedure: Approval from Institutional Ethics Committee was taken before starting the study. All health care professionals and students were contacted personally. The study was explained to them in brief. A predesigned questionnaire was provided to them which consisted of ten questions for assessment of awareness, attitude and practice of pharmacovigilance and ADR reporting. Consent of participants was taken in written informed consent form. They were asked to fill the questionnaire without any assistance. It required approximately 10 to 15 minutes filling the questionnaire. Out of total 799 participants who were asked to fill the questionnaire, 565 (70.7%) responded to the questionnaire. There were 11 pharmacists, only 2 responded. Therefore, pharmacists were not included for analysis. Thus total 563 questionnaires were analyzed. All questionnaires were completely filled. Data collected were analyzed and percentages were calculated. Awareness between healthcare professionals and students was compared using chi square test. Practice of ADR reporting and reasons for nonreporting were assessed only for healthcare professionals as Pharmacovigilance Programme of India (PvPI) mentions only healthcare professionals who can report the ADR. [6]
RESULTS AND DISCUSSION
Among healthcare professionals, 73% (73/100) doctors and 57% (115/202) nurses responded to the questionnaire. Among students, 72.4% (210/290) MBBS students, 96% (100/104) pharmacy students, 61% (33/54) nursing students and 84.2% (32/38) physiotherapy students responded to the questionnaire. 65.4% of healthcare professionals (83.5% of doctors and 53.9% of nurses), while 83.5% of students have ever heard of the term “Pharmacovigilance”. (Table 1) Awareness about this term was different between the two groups with statistical significance (p value < 0.001). 41.5% of healthcare professionals (58.9% of doctors and 30.43%of nurses) and 39% of students were able to define Pharmacovigilance. (Table 2) No statistically significant difference between the two groups (p value > 0.005) was found. 39.4% of healthcare professionals (43.8% of doctors and 36.5% of nurses) and 31.7% of students were aware of National Programme for Pharmacovigilance, (Table 2) with no statistically significant difference (p value > 0.005). 53.7% of healthcare professionals (60.3% of doctors and 49.6% of nurses) and 57.1% of students had the knowledge of ADR reporting. (Table 2) No statistically significant difference (p value > 0.005) was found. The awareness that any healthcare professional can report ADR was present in 43.1% of healthcare professionals (67.1% of doctors and 27.8% of nurses) and 48.3% of students, (Table2) with no statistically significant difference (p value > 0.005). 72.3% of healthcare professionals (91.8% of doctors and 60% of nurses) and 60.8% of students were aware how to report ADR. (Table 2) There was statistically significant difference between two groups with p value < 0.01. 78.1% of doctors and 62.6% of nursing staff have not reported any ADR to any authority in last two years. Only 21.9% of doctors and 37.4% of nursing staff have reported any ADR to any authority in last two years. (Figure 1) About 56% of doctors and 46% of nurses reported lack of awareness about the reporting system as the major cause of non-reporting of ADR. 20.5% of doctors and 23.5% of nurses were not sure about the reason for non-reporting. (Table 3) 68% of healthcare professionals (75% of doctors and 63% of nurses) and 76.8% of students opined that ADR reporting is very important. Only 2.6% of healthcare professionals and 1.6% of students considered it as waste of time. (Figure2) 84% of healthcare professionals (97% of doctors and 75.6% of nurses) and 84.3% of students accepted that pharmacovigilance should be an active part of medical curriculum. Only 5.8% of healthcare professionals and 6.4% of students denied. (Figure 3) The overall awareness about pharmacovigilance and ADR reporting was low, both in healthcare professionals and students. Though majority of students (83.5%) had heard the term “Pharmacovigilance”, less than half (39%) could define it. These students are told about it in pharmacology but not actively discussed. This shows that there is a need to stress on pharmacovigilance during undergraduate teaching. On the other hand 65% of healthcare professionals had heard this term which is significantly lower than that of students, but only 41% could define it. More doctors (59%) were able to define than nursing staff (30%). Healthcare professionals are not exposed to pharmacology after II year of undergraduate course, which may be responsible for their low awareness about the term “Pharmacovigilance”. A Nigerian study done on community pharmacists also reported similar results in which 55% of responders were aware of the term “pharmacovigilance”, but only 18% could define it. [8] More alarming was the lack of awareness about the national programme. More than half of healthcare professionals and students were not aware about the national programme which is running since 2004 in India. If they do not know about it, how could they be expected to participate in it? To make the national programme successful, it is important to aware the healthcare professionals about it and how it functions. This can be done by awareness campaigns, information leaflets etc. About half of healthcare professionals (54%) and students (57%) were aware about ADR reporting. The awareness that every healthcare professional including doctor, nurse, pharmacist and physiotherapist can report ADR was very low, which is in accordance with the findings of Gupta P et al.[13] This awareness was minimum among nurses, both nursing staff (28%) as well as nursing students (21%). Active involvement of paramedical staff in spontaneous reporting is very important, since they are in close contact with the patients and for a longer duration as compared to doctors. The ultimate goal of Pharmacovigilance is that the benefits of medicine use outweighs the risks and thus safeguard the health of patients. Spontaneous ADR reporting by healthcare professionals can play an important role to achieve this goal. In present study only about 22% of doctors and 37% of nurses had reported ADR to any authority in last two years. Other studies have also reported such low levels of ADR reporting. [7-9] Underreporting is a major and worldwide problem associated with spontaneous reporting system. A systematic review by Hazell L et al has stated significant and widespread under-reporting of ADRs to spontaneous reporting systems including serious or severe ADRs. [10] Aggressive interventions are required to encourage the healthcare professionals for ADR reporting. They need to be informed that why ADR reporting is necessary and how it can help in ensuring safe and rational use of medicines. Prescribers can be encouraged by providing feedback on their ADR reports, discussion on ADR reports in academic meetings and publishing bulletins on ADRs. Oreagba et al have suggested that remuneration for ADR reporting may increase the reporting rate. [8] Lack of awareness about the reporting system was the most common reason stated by responders (56% of doctors and 46% of nurses) for nonreporting. This finding is in accordance with that reported by other studies, [8, 9, 13] and is understandable as there is no well established reporting system in this hospital and healthcare professionals are not well aware about the national programme. Thus to improve the reporting rate, ADR monitoring centers should be established in all healthcare institutions, at least at tertiary care level. The recent Pharmacovigilance Programme of India targets to establish such centre in most of the Medical Colleges of India in coming years. [6] This may help to improve the existing scenario. Further to increase the awareness about ADR reporting system, regular training sessions and awareness campaigns should be conducted which may help in improving ADR reporting rate. Ramesh M et al have reported 63% increase in ADR reporting in one year after the launch of continuous awareness campaign. [14] As mentioned by other studies lack of time, complex procedure and non significant ADR, were among the other reasons for non-reporting in present study also. [8, 13] Both healthcare professionals and students had positive attitude toward ADR reporting and majority of them considered it as very important. Only 2.6% of healthcare professionals and 1.6% of students considered it as waste of time. Further majority of responders (84%) opined that pharmacovigilance should be taught as an active part of medical curriculum. As medical students are future healthcare professionals, there is a need to actively train them, so that pharmacovigilance becomes a part of their medical practice. As recommended by Rehan et al knowledge and awareness of pharmacovigilance among prescribers can be improved by a reinforcement training programme at the commencement of internship and thereafter through continuous education programmes. [9] The major limitation of the present study is that the study findings could not be applied to wider medical community as the study was restricted to hospital setup. Therefore it is recommended that several studies of similar kind especially in community setup need to be conducted to know the awareness and attitude of healthcare professionals in community and their practice of pharmacovigilance. This will help to find out the present status and to develop strategies to improve the ADR reporting system in India.
CONCLUSION
The results of present study show lack of awareness about pharmacovigilance in healthcare professionals and students. ADR reporting rate is also very low among healthcare professionals. There is a great need to create awareness and promote ADR reporting among healthcare professionals. Regular training sessions to stress the importance of ADR reporting and the functioning of reporting system are required. Further awareness about the national programme need to be increased, this will ensure greater participation from healthcare professionals and success of such programmes. Pharmacovigilance should be a part of undergraduate curriculum of not only MBBS students but also of students of nursing, physiotherapy and pharmacy. As these students are future healthcare professionals, this will help in developing a culture of ADR reporting in the country.
Englishhttp://ijcrr.com/abstract.php?article_id=2297http://ijcrr.com/article_html.php?did=22971. Tripathi KD. Essentials of medical pharmacology. 6th ed. New Delhi: Jaypee Brothers Medical Publishers (P) Ltd; 2010. p. 79.
2. Classen DC, Pestonik SL, Evans RS, Lloyd JF, Burke JP. Adverse drug events in hospitalized patients. JAMA 1997;277(4):301-6.
3. Lazarou J, Pomeranz BH, Corey PN. Incidence of adverse drug reactions in hospitalized patients – a meta-analysis of prospective studies. JAMA 1998;279:1200-5.
4. Oates JA. The science of drug therapy. In: Brunton LL, editor. Goodman and gilman?s the pharmacological basis of therapeutics. 11th ed. New York: McGraw-Hill; 2006. p. 133-5.
5. List of NPP centers [Online]. [2010?] [cited 2011 Jan 22]; Available from: URL:http://www.pharmacovigilance.co.in/nppc entreslist.html
6. Pharmacovigilance programme of India for assuring drug safety. Central drug standard control organization, Directorate General of health services, Ministry of health and family welfare, Government of India [Online]. [cited 2011 Aug 8]; Available from: URL: http://cdsco.nic.in/pharmacovigilance.htm
7. Subish P, Mohammed Izham MI, Mishra P. Evaluation of the knowledge, attitude and practice on adverse drug reactions and pharmacovigilance among healthcare professionals in a Nepalese hospital: a preliminary study. The Internet Journal of Pharmacology 2008;6(1).
8. Oreagba IA, Ogunleye OJ, Olayemi SO. The knowledge, perceptions and practice of pharmacovigilance amongst community pharmacists in Lagos state, south west Nigeria. Pharmacoepidemiol Drug Saf 2011;20:30-5.
9. Rehan HS, Vasudev K, Tripathi CD. Adverse drug reaction monitoring: knowledge, attitude and practices of students and prescribers. Natl Med J India 2002;15:24-6.
10. Hazell L, Shakir SA. Under-reporting of adverse drug reactions : a systematic review. Drug Saf 2006;29(5):385-96.
11. Xu H, Wang Y, Liu N. A hospital-based survey of healthcare professionals in the awareness of pharmacovigilance. Pharmacoepidemiol Drug Saf 2009 Jul;18(7):624-30.
12. Ohaju-Obodo JO, Iribhogbe OI. Extent of pharmacovigilance among resident doctors in Edo and Lagos state of Nigeria. Pharmacoepidemiol Drug Saf 2010;19(2):191- 5.
13. Gupta P, Udupa A. Adverse drug reaction and pharmacovigilance: knowledge, attitude and perceptions amongst resident doctors. J Pharm Sci and Res 2011;3(2):1064-9.
14. Ramesh M, Parthasarathi G. Adverse drug reactions reporting: attitude and perceptions of medical practitioners. Asian J Pharm Clin Res 2009;2(2):10-4.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareINFLUENCE OF MICROBIAL INOCULATION ON PLANT HEIGHT OF BLUE PINE (PINUS WALLICHIANA) AND HIMALAYAN CYPRESS (CUPRESSUS TORULOSA) UNDER NURSERY CONDITIONS
English1522Malik A. AEnglish Zargar M .YEnglish Najar G. REnglish Mir S. A Agha FEnglishA pot experiment was carried out during 2009 -2010 to study the impact of microbial inoculants on plant
height of Blue pine (Pinus wallichiana A.B. Jackson) and Himalayan cypress (Cupressus torulosa Don) under nursery conditions. The experiment was laid in Completely Randomized Design with three replications which comprised forty-two treatment combinations of seven inoculants (Azotobacter sp., Azospirillum sp., Pseudomonas fluorescens, Bacillus subtilis, Pisolithus tinctorius, Laccaria laccata and control). The growth character viz., plant height at various intervals responded significantly to all the microbial inoculants. Maximum plant height was recorded in Himalayan cypress (38.72 cm) as compared
to Blue pine seedlings (24.78 cm). Among microbial inoculants the two ectomycorrhizae viz., Pisolithus tinctorius and Laccaria laccata gave best results than rest of the inoculants. It was followed by Azotobacter sp., Azospirillum sp., Pseudomonas fluorescens and Bacillus subtillis. Thus the two treatments viz., Pisolithus tinctorius and Laccaria laccata proved to be the best for the studied parameter
viz., plant height of both the species.
EnglishPinus wallichiana, Cupressus torulosa, Microbial inoculation, Azotobacter, Azospirillum, Pseudomonas, Bacillus, Pisolithus, LaccariaINTRODUCTION
The geographical area of Jammu and Kashmir is 1,01,387 km2 excluding the area under Pakistan and China with recorded forest area of 20,230 km2 inside line of control. However, recorded forest cover as per Forest Survey of India is only 16,309 km2 (Anonymous, 2009a). Commercial forests of the state occupy only 8,26,939 ha (Anonymous, 2005). As per inventory records, very dense forests in Jammu and Kashmir occupy 2,95,800 ha, moderately dense 6,50,700 ha, open forests have now spread over 6,84,400 ha and rest of the recorded forests have been engulfed by blanks and scrubs (Anonymous, 2009b). The growing stock of our commercial forests is 132.9 million m3 with average annual yield of 1.65 m3 ha-1 (Anonymous, 2009b). With this productivity annual yield of timber from commercial forest area alone must be 27.25 million cubic feet. Contrary to this fact, we presently import timber. Due to timber mining more than 60 per cent of our demarcated forests have been declared as uncommercial/degraded (Anonymous, 2005). Natural regeneration does not practically take place in forests where crown density is less than 40 per cent. Relying on natural succession, it will take us hundreds of years to regenerate the degraded forests to climax stage with species like Pinus wallichiana Jackson (kail), Cedrus deodara (Roxb.), G. Don (Deodar), Abies pindrow Spach (silver fir), Picea smithiana Wall. (spruce) and Cupressus torulosa Don (Himalayan cypress) which dominate vegetation of our forests.To meet the huge demand and supply of timber, fuelwood and firewood, raising of blue pine and Himalayan cypress forests on degraded forest patches can be a good and viable option in future. The indiscriminate use of inorganic fertilizers and pesticides is neither environmentally safe nor economically feasible. There is pressing demand for microbial inoculants for quality seedling production in nursery and also the establishment of plantation to increase the forest productivity. Bioinoculants are cost effective, ecofriendly, cheaper and renewable sources of plant nutrients and play a vital role in maintaining long-term soil fertility and sustainability. Thus, to meet the challenges like poor regeneration, deforestation and spread of wastelands, introduction of microbial inoculants at the nursery stage of forest trees has become inevitable. Although various aspects of mycorrhizal impact of the forest trees have been studied, no work has been done on the impact of other microbial inoculants on the regeneration of forest trees. Therefore, the present study was undertaken to determine the role of microbial inoculation on growth attribute viz.,plant height of Blue pine and Himalayan cypress under nursery conditions. MATERIALS AND METHOD The present investigations were undertaken at the Forest Nursery of Department of Forestry, Faculty of Agriculture and Regional Research Station, SKUAST-Kashmir, Wadura, Sopore during 2009- 2010.Microbial inoculants isolated from rhizosphere of blue pine and Himalayan cypress forest stands were used in the studies. Mass production of microbial inoculants The two free living aerobic nitrogen fixing bacteria viz., Azotobacter sp. and Azospirillum sp. were mass cultured using nutrient medium enriched with glucose and peptone. Plant growth promoting rhizobacteria (PGPR) viz., Pseudomonas fluorescens and Bacillus subtilis were mass propagated in King?s B nutrient broth. The two ectomycorrhizae viz., Pisolithus tinctorius and Laccaria laccata were mass multiplied in Melin Norkran?s nutrient broth and Potato Dextrose Agar, respectively. Field operations For the microbial inoculation, one year old seedlings of blue pine and Himalayan cypress of uniform heights and collar diameter growing in polyethylene bags (9? x 7?) containing 1 kg potting material of soil and sand mixture in the ratio of 1:1 were selected. Microbial inoculation For inoculation, the different broth cultures of Nfixers, P-solubilizers and ectomycorrhizal inoculants isolated from local forest stands were applied to the potting material (25 ml/seedling) in the month of March, 2010, without disturbing the root system of the seedlings. Nursery operations The seedlings were irrigated with rose-cans as and when needed and maintained virtually weed free by manual weeding Plant growth measurement The growth parameter viz., plant height (cm) of both the species were measured by using measuring tape at an interval of 2 months upto 12 months.The plant height of the seedlings at the initial stage of the experiment were also recorded. Statistical analysis The data was statistically analysed by using O.P Stat software developed by Haryana Agriculture University, Hisar.
RESULTS AND DISCUSSION Plant height The data on impact of various microbial inoculants on plant height of Blue pine seedlings indicates that mean plant height was significantly more in response to various treatments as compared to control (Table-1;Fig 1, Plate-1). Azotobacter and Azospirillum inoculation exhibited 37.17 and 36.83 per cent more plant height over control. Similarly Pseudomonas flourescens and Bacillus subtilis inoculation resulted in 35.56 and 33.91 per cent more plant height while as the inoculation with two ectomycorrhizal fungi viz., Pisolithus tinctorius and Laccaria laccata resulted in 42.97 and 40.77 per cent more plant height as compared to control respectively. However, the application of Pisolithus tinctorius showed maximum increase (42.97%) in plant height over control, thus proved superior over all the individual inoculants. Moreover, there was an increasing trend in plant height from April to October and from December onwards till February there was a slight increase. The interactions between inocula and months were significant till October and from December to February it was non-significant. Perusal of the data presented in Table-2,Fig 2,Plate-1 shows that the application of various microbial inoculants significantly enhanced the mean plant height of the Himalayan cypress seedlings as compared to control. Amongst various microbial inoculants, Pisolithus tinctorius resulted in maximum increase in plant height over control (38.27 %). It was followed by Laccaria laccata (35.66%), Azotobacter sp. (28.29%), Azospirillum sp. (25.91%), Pseudomonas fluorescens (21.76%) and Bacillus subtilis (19.36%), respectively. Treatment of seedlings with ectomycorrhizal fungi viz., Pisolithus tinctorius was significantly superior over all other treatments. Plant height revealed a significant increase from April to October and from October on wards till February there was a slight increase. Moreover, the interactions between inocula and month?s were significant till October and thereafter it was non-significant. The increase in shoot height by P. tinctorius and L. laccata could be attributed to the production of growth promoting substances like auxins (Dehn, 1982) and enhancement of water absorption and nutrient mobilization (Dar et al., 1997) by vastly increased surface area network of the fungal mycelia (Myer, 1992). In case of Azotobacter and Azospirillum sp. inoculation the increase in shoot height could be ascribed to nitrogen fixing ability, synthesis of growth promoting substances like cytokinens, gibberellins, auxins (Reynders and Vlassak, 1979; Hartmann et al., 1983; Jain and Patriquin, 1985) and production of antifungal antibiotics (Chahal and Chahal, 1988). However, increase in shoot height by inoculation with Pseudomonas fluorescens and Bacillus subtilis could be through iron chelating siderophores (Schippers, 1988) by releasing phytohormones, solubilizing P and reduction in population of deleterious microorganisms (Weller, 1988). Further our findings are in close conformity with the results of Oh and Park (1989), Jeffries and Dodd (1991), Natarajan et al. (1995) who reported that P. tinctorius and L. laccata inoculation resulted in enhancement of plant height of Acaccia nilotica, Quercus serrata, Eucalyptus camaldulensis and E. deglupta seedlings respectively. similarly, the enhancement in plant height with respect to Azotobacter and Azospirillum sp. has also been reported in Quercus serrata (Pandey et al., 1986) in peach (Awasthi et al., 1996). Moreover, the inoculation of clover plants with Pseudomonas putida has also been reported to enhance the plant height (Meyer and Linderman, 1986). However, the maximum increase in shoot height of Himalayan cypress seedlings lies in the fact that Himalayan cypress being a fast growing species, has got an efficient root system as compared to kail which is comparatively a slow growing species. Moreover, the gradual decline in plant height of both the species in the later half of study period could be due to below freezing soil temperatures and short growing season of conifers.
Englishhttp://ijcrr.com/abstract.php?article_id=2298http://ijcrr.com/article_html.php?did=22981. Anonymous, 2005. Handbook of Forest Statistics. Jammu and Kashmir Government; Forest Department, Srinagar, JandK, pp 10. 2. Anonymous, 2009a. Forest Survey of India. Indian State of Forest Report 2009, 5 : 44. 3. Anonymous, 2009b. Digest of Forest Statistics. Jammu and Kashmir Government; Forest Department, Srinagar, Jammu and Kashmir. 4. Awasthi, R.P., Godara, R.K. and Kainth, N.S. 1996. Interaction effect of VA-mycorrhizae and Azotobacter inoculation on peach seedlings. Indian Journal of Horticulture 53(1) : 8-13. 5. Chahal, P.P.K. and Chahal, V.P.S. 1988. Biological control of root-knot nematode of brinjal with Azotobacter chroococcum. In : Advances in Plant Nematology [Eds. M.A. Maqbool, 6. A.M. Golden, A.M. Gaffar and A. Krusberg]. Research Centre, University of Karachi, pp. 257-263. 7. Dar. G.H., Zargar, M.Y. and Beigh, G.M. 1997. Biocontrol of Fusarium root rot in common bean (Phaseolus vulgaris L.) by using symbiotic Glomus masseae and Rhizobium leguminosarum. Microbial Ecology 34 : 74-80. 8. Dehn, H.W. 1982. Interaction between vasicular-arbuscular mycorrhizal fungi and plant pathogens. Phytopathology 72 : 115-119. 9. Hartmann, A., Singh, M. and Klingmuller, W. 1983. Isolation and characterization of Azospirillum mutants excreating high amounts of indoleacetic acid. Canadian Journal of Microbiology 29 : 916-923. 10. Jain, D.K. and Patriquin, D.G. 1985. Characterization of a substance produced by Azospirillum which causes branching of wheat root hairs. Canadian Journal of Microbiology 31 : 206-210. 11. Jeffries, P. and Dodd, J.C. 1991. The use of mycorrhizal inoculants in forestry and agriculture. In : Handbook of Applied Mycology. [Eds D.K. Arora, Bharat Raj, K.G. Mukerji and G.R. Knudsen]. Marcel Dekker, New York, USA, pp. 155-185. 12. Meyer, J.R. and Liderman, R.G. 1986. Response of subterraneum clover to dual inoculation with VAM fungi and a plant growth promoting bacterium Pseudomonas putida. Soil Biology and Biochemistry 18 : 185-190. 13. Myer, M. 1992. Mycorrhizas, their use as biofertilziers. The horticulturists 2 : 8-12. 14. Natarajan, K., Nagarajan, G. and Reddy, M.S. 1995. In vitro mycorrhization and growth response of Acacia nilotica seedlings by inoculation with ectomycorrhizal fungi. Indian Journal of Microbiology 35 : 35-38. 15. Oh, K.I. and Park, W.S. 1989. The effect of ectomycorrhizae and nitrogen levels on the growth of Quercus serrata seedlings. Journal of Korean Forestry Society 78 : 160-167 16. Pandey, P.K., Bahl, R.K. and Rao, P.R.T. 1986. Growth stimulating effects of nitrogen fixing bacteria (biofertilizer) on oak seedlings. Indian Forester 112(1) : 75-79. 17. Reynders, L. and Vlassak, K. 1979. Conversion of tryptophan to indoleacetic acid by Azospirillum brasiliense. Soil Biology and Biochemistry 11 : 547-548. 18. Schippers, B. 1988. Biological control of pathogens with rhizobacteria. Phiols. Trans. R. Soc. Land. B. 318 : 283-292. 19. Weller, D.M. 1988. Biological control of soil borne plant pathogens in the rhizosphere with bacteria. Annual Review of Phytopathology 26 : 379-407.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareAN OBSERVATIONAL STUDY ON SENSORY BASED OUTDOOR PLAY PREFERENCES IN CHILDREN AGED BETWEEN 3 - 12 YEARS : A PRELIMINARY STUDY
English2330KR. BanumatheEnglish Vineeta .I. RamEnglish Alzeena PintoEnglish Samuel JonathanEnglishAim: To find sensory based outdoor play preferences of children between the ages 3 -12 years. Method: 90 normal children were observed by 3 raters for 30 minutes minimum, while engaged in free play at outdoor playgrounds. Play preferences were observed for the affinity towards certain sensory components; tactile, auditory, vestibular and proprioception. The most observable sensory based play preferences were noted for quality and frequency and the children were scored on a 7 point Likert scale. The data was then grouped by age into 3 groups, 1, 2 and 3; 3.1 to 6 years, 7.1 to 9 years and 10.1 to 12 years respectively and analyzed using SPSS, version 16. Results: On analysis of data between the groups showed that there was significant difference at p0.05 levels. Auditory based play preferences showed a significant difference at p0.05 levels and significant difference between the rest of the components at pEnglishPlay preferences, Sensory Based Play, Outdoor Play PreferencesINTRODUCTION
Play is a child?s primary and most important occupation1 .It has historically been regarded by occupational therapists as both an indicator of development and a means of intervention2 . Many occupational therapists feel that the common themes in play include intrinsic motivation, internal reality, and internal locus of control, are needed for a child to engage in playful behaviors and interactions. When these are present, a child is self-motivated to engage in a play activity, is free fromData Analysis rules, procedures or guidelines to follow during the play, and is able to self-direct play3 . If a child has a deficit in his or her ability to be intrinsically motivated, to suspend reality, to have an internal locus of control, to be happy, energetic or playful, or has an inability to process sensory information from play experiences, then a child?s development may be stifled4 . Taking this into consideration, how children make play choices and assign meaning to the experience of this occupation is an important area of study for occupational therapists, to better understand this occupation and the use of play within practice5 . Significant research has been accumulated on play preference with regard to gender and age; less research exists on the relation of play preferences to the ability4 .Research on the meaning of play for children or on children?s perspective and rationale for their play choices remains scarce6 . A study on Children?s perceptions of play experiences and the development of play preferences, found that it would be beneficial for therapists to understand the long-term implications of play choices in children and their impact on development over time. Also suggested the relationship between a children?s sensory processing and his or her specific play choices could be an important area for further study4 . Based on this, and related literature, this preliminary study was undertaken to further understand, explore and describe the relationship between sensory processing and play preferences which in turn will improve the use of play in pediatrics evaluation and intervention with the following research question: Do children between the ages 3 and 12 years have sensory based play preferences.
Aim
The aim of this study is to find sensory based outdoor play preferences of children between 3 and 12 years of age.
Objectives
To observe children between 3 -12 years playing in an outdoor playground.
To identify the sensory based play preferences among these children.
Research Hypothesis
Null Hypothesis
There will not be any significant difference in sensory based play preferences for children between the ages 3 and12.
Methodology
Pilot Study
A pilot study was conducted initially to check if sensory based play preferences are present and can be assessed. It is also to assess the reliability of scores of the three raters. This was done by each rater observing the same five children individually in an open playground with common play equipment such as swings, slides; see saw, monkey bars, the merry go round and sand pits for duration of 30 minutes each. Observations were noted by the 3 raters separately and were scored using the 7 point Likert scale.
Inclusion Criteria
Normal children of both boys and girls aged between 3 to 12 years.
Exclusion Criteria
Children with any physical or mental disability.
Children who played for less than 30 minutes.
Setting
Two outdoor playgrounds with similar types of play equipment such as slides, swings, see-saw, monkey bars, sand pits and merry go rounds.
Sample Size
90 children
Study Design
Observational Study
Tool
A stop watch
Scoring Criteria
7 point Likert's scale
Procedure
Children who came with their parents were selected randomly for the study; oral consent was taken prior to the study from the children?s parents along with the child's details demographic, physical and mental health.
Three investigators directly observe the spontaneous play of an individual child in a playground for 30 minutes minimum from a suitable vantage point with the children being unaware of the observers.
Using the 7- point Likert scale as the tool of measure, each observer evaluated the same individual child independently using the definitions mentioned before in the Operational definitions as a reference.
The average of each observer?s score for each variable was taken following which the data was analyzed.
Data Analysis
The results were analyzed at p0.05 levels. The Post HOC revealed no significant difference between the groups 1 and 2 and 3 at p>0.05 levels.
Play preferences within group 1
The mean scores obtained for tactile based play preferences 6.1892,±.84452,auditory based play preferences 5.3514,±.75337,proprioceptive based play 6.3243,.70923 and vestibular based play preferences 6.2937,.61756.We used ANOVA to find the significant difference between these mean values, with the „F? score being 14.663,we found that they were significantly different at the p0.05 levels.
Play preferences within group 2
The mean scores obtained for tactile based play preferences5.3226,±.79108,auditory based play preferences 4.8065,±.47745,proprioceptive based play 6.8710,.34078 and vestibular based play preferences 6.3548,±.66073.We used ANOVA to find the significant difference between these mean values, with the „F? score being 72.287,we found that they were significantly different at pEnglishhttp://ijcrr.com/abstract.php?article_id=2299http://ijcrr.com/article_html.php?did=22991. Bundy AC. Play Theory and Sensory Integration. In: Bundy AC, Lane SJ, Murray EA editors. Sensory Integration Theory and Practice. 2nd ed. Philadelphia: FA Davis; 2002. p. 227-40.
2. Parham L, Primeau L. Play and occupational therapy. In: Parham LD, Fazio L, editors. Play in occupational therapy for children. St. Louis: Mosby; 1997. p. 02–21.
3. Morrison CD, Metzger P. Play. In: Schrefer J, White K, Mosby L, editors. Occupational Therapy for Children. 4th ed. St. Louis: Mosby; 2001. p.528-40.
4. Miller E, Kuhaneck H. Children?s perceptions of play experiences and the development of play preferences: A qualitative study. American Journal of Occupational Therapy 2008; 62:407-15.
5. Couch KJ, Deitz JC, Kanny EM. The role of play in pediatric Occupational therapy. Am J of Occupational Therapy1998; 52: 111–17.
6. Smith CJ, Kuhaneck H. Play preferences of typically developing children and children with developmental delays between the ages 3 and 7 years. Occupational Therapy J of Research 2008; 1:19-20.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesTESTING INDEX VOLATILITY OF INDIAN STOCK MARKET IN THE CONTEXT OF FOREIGN INSTITUTIONAL INVESTOR"S INVESTMENT
English3146Himanshu BarotEnglish Dr. V.K.SapovadiaEnglishIndian stock market has seen an unprecedented growth in the last few years. Since year 2002, Indian market has grown from a much volatile conditions to growth phenomena; this has been due to not only
the domestic market but also the international investors. FIIs investment is considered to be one of the key influencing factors after the economic fundamentals. From 1993 reforms in the Indian Capital market on liberalisation of the FII flows has a considerable impact on Indian stock market. The main objective of the study is to test index volatility of Indian stock market in the contest of FII?s net investment in equity and for this two net change in indices have been considered i.e. BSE Sensex and SandP CNX Nifty on annual basis from January 2000 to December 2011. The empirical result found through statistical tests like correlation, regression and t-test which reveals that significant relation between all three variables and result found that 2008 and 2011 were one of the worst years for Indian markets which make it the worst performer in Asia. Global financial recession in 2007 and slowing growth, rising inflation, and policy paralysis have blown the Indian markets off course in 2011. In 2008, the sub-prime crisis and Lehman brothers collapse crippled the global markets when 2011 saw high inflation rate dampening the market spirit. The Great Indian Dream seems to be coming to a close as most of the FIIs are pulling out of Indian markets. The FIIs were net sellers in year 2008 and 2011, pulling out their positions form the Indian market which leads to conclude that the FII is one of the key movers on volatility of Indian stock market and FII?s investment is not negligible for Indian Capital Market.
English1.INTRODUCTION
A well-developed stock market has its impact on the development of economy. It provides investors with an array of assets with varying degree of risk, return and liquidity. This increased choice of assets and the existence of a vibrant stock market provide savers with more liquidity and options, thereby inducing more savings. Increased competition from foreign financial institutions also paves the way for the derivatives? market. All this, according to the mainstream belief, encourages more savings in equity related instruments. This, in turn, raises the domestic savings rate and improves capital formation.
Above model indicating that portfolio investment is also a stimulus of economic development because, it?s a main source of fund of corporate. The demand of portfolio investment is created by companies and their routes are decided by government. It is considered as less reliable source of fund for economic development because it fluctuates on some minor trends of economy.
2 Foreign Institutional Investors
FII or the Foreign Institutional Investors are basically referred to investors who are organized in the form of an institution or entity and indulge in investing funds in the financial market of a foreign country, i.e. different from where the entity was originally registered or incorporated. In India, FII can invest their funds in the country only under the norms prescribed by Security and Exchange Board of India (SEBI). FII?s investment includes mutual funds, investment trust, asset management company, nominee company, bank, university funds, endowments, foundations, charitable trusts, charitable societies, overseas pension funds etc. It was in July 1991 that the New Economic Policy was unveiled that ushered in an era of Liberalization Privatization and Globalization (LPG) for the Indian economy. In September 1992 and 1993, India opened its stock market for foreign investors, which facilitated receipt of funds from foreign institutional investors in the form of equities. The enactment led to sweeping changes where various restrictions imposed on investments by the foreign investors in India were eased. SEBI opened a new path for the foreigners to invest in India by simplifying many terms and conditions due to which a large number of foreign investors flocked towards India.
2.1 Number of Foreign Institutional Investors (FIIs)
One of the most important features of the development of stock market in India in the last 20 years has been the growing participation of FIIs. Since September, 1992 when FIIs were allowed to invest in India, the no. of FIIs has grown over a period of time. The net addition in SEBI registered FIIs failed to keep up the momentum seen in 2007-08 and 2008-09 wherein there was addition of 322 and 316 FIIs respectively. There was a net addition of 78 SEBI registered FIIs in 2009-10 which took their total number to 1,713 at end March 2010 compared to that of 1,635 at the end of March 2009 (Table I and Chart II).
2.2 Trends in Investment of Foreign
Institutional Investors
This is not unusual as most of the developing economies might be experiencing the same patterns. The increasing role of FIIs has brought in the development of our stock markets as well such as expansion of the business in securities, increased depth and breadth of the market, etc. FIIs contribute to almost 13% of the entire market capitalization at National Stock Exchange in India. If we talk of FIIs investment, this has been continuously grown over years except 1998-99 and 2008-09 when FIIs sold more than they purchased in Indian stock market. (Table II) The gross purchases of debt and equity by FIIs increased by 17.27 percent to Rs. 9,92,599 crore in 2010-11 from Rs. 8,46,438 crore in 2009-10 (Table II). The combined gross sales by FIIs increased by 20.23 percent Rs. 8,46,161 crore from Rs. 7,03,780 crore during the same period in previous year. The total net inflow of FII was increased by 2.65 percent Rs. 1,46,438 crore in 2010-11 as against an inflow of FII was Rs. 1,42,658 crore in 2009-10. This was the highest net inflow for any financial year so far.
Cumulative investment by FIIs at acquisition cost, which was USD 89.3 billion at the end of March 2010, increased to USD 121.5 billion at the end of March 2011 (Figure 2). Because of their war chests of money, the role of FIIs can?t be ignored. FIIs have dynamic portfolios across countries which they use to restructure and rebalance depending on the market conditions, definitely, with a motive to increase their gains. Because of their size of investment in any market, they have the ability to make or break the fortunes of any market.
2.3 Foreign Institutional Investments- Equity and Debt
FIIs were allowed to invest in the Indian Capital Market from September 1992. Investments by them, however, were first made in January 1993. Till December 1998, investments were related to equity only as the Indian gilts market opened up for FII investment in April 1998. FIIs? investment in debt started from January 1999. Foreign Institutional Investors (FIIs) continued to invest large funds in the Indian securities market. For two consecutive years in 2004-05 and 2005-06, net investment in equity showed year-on-year increase of 10%.
After experiencing a net outflow of Rs 477,070 million in equity instruments in 2008-09, FIIs scaled record equity investment in 2010-11 which stood at Rs 1,105,297 million. (Table III) The net investments by FIIs in the debt segment also bounced back in 2010-11 with a staggering all-time high of Rs 421,451 million compared to that of Rs 18,950 and Rs 324,380 in 2008-09 and 2009-10 respectively.
3 Review of Literature
Many empirical studies have been conducted to examine the relationship between stock prices and buying of equity by FIIs in Indian stock market. Purnendra Verma (2001)2 studied the impact of FII on Indian Capital Market from 1993 to 2001 and found that there is significant effect of FIIs on Nifty and no significant effect on BSE Sensex, but in both the cases the co-efficient of correlation is low and very low so the effect are less and very less respectively. Study concluded that FII did not have any significant impact on the Indian Capital Market. T. N. Aravind, et al. (2008)3 examined the study on FII?s influence on Indian stock market for the period of 2003 to 2008 and concluded that in Oct. 2007, speculation about governments plan to control P-Notes had caused the biggest fall in Indian stock market. And they have proved that there is a direct relationship between the FII?s money flow and the movement of Sensex. Prasanna P.K. (2008)4 examined the contribution of FIIs in SENSEX base companies for the period of 2001 to 2006 and found that foreign investors invested more in companies with a higher volume of shares owned by the general public. The promoters? holdings and the foreign investments are inversely related. Study argued that the foreign institutional investors withdraw their money when the stock market performance starts slowing down. Dr. Rahul Singh, et al. (2008)5 studied of FIIs investment flow and SENSEX movement for period from January, 2004 to December, 2007 and proved with empirically that there is a negative correlation between the net FIIs and volatility in SENSEX. And with respect to net FIIs inflows and returns on SENSEX have a positive and significant correlation. Soumita Patra, et al. (2009)6 conducted the study on impact of FIIs flow on the BSE SENSEX and Nifty between 1997 and 2008 and proved that there is no significant relationship between FII and SENSEX excluding years 2003 and 2004 where FII and Nifty there is no significant relation. Gaurav Agrawal, et al. (2010)7 investigated the causal relationship between Nifty and FIIs net investment for the period January, 1999 to February, 2009 using daily data and through application of correlation test, found that Nifty is positively correlated to FIIs. Madan Sabnavis, et al. (2010)8 analyzed implications of FIIs inflow in Economic Studies from January to September 2010 and empirically proved that the coefficient of correlation is quite strong between FIIs flows and SENSEX movement which means there is a significant relationship. A.Q.Khan and Sana Ikram (2010)9 tested the efficiency in relation to the impact of FIIs largely on the Indian Capital Market during the period 2000 to 2010 and proved that there is a significant relation between the movement of FII and the two major stock exchanges of India that is the BSE and NSE. However the coefficient of correlation is a low degree of positive correlation indicating that the effect is not very strong. The study concluded that the FIIs investment has a significant impact on Indian Capital Market and the Indian Capital Market is Semi-strong form efficient in relation to the impact of FIIs investment on Indian Capital Market. Narendra Singh Bohra and Akash Dutt (2011)10 studied data from 2000 to 2009 and found that there is a positive correlation between stock market and investment of FII?s in a relation that Sensex follows the investment behavior of FII?s and in the case of individual group securities FII?s had shown a positive correlation in less regulated and high capitalized securities in the market to earn high equity yield. And study suggest to the policy implication that the authorities can focus on domestic economic policies to stabilize the stock market. M. Anuradha Reddy (2011)11 examined the FIIs investment behavior and its relationship between SENSEX movement during years 2000 to 2011 (May, 31) and found that the FIIs are influencing the Sensex movement and proved evidently there is a significant relation between FIIs flow and Sensex movement. Ravi Akula (2011)12 conducted the study on trends in foreign institutional investment in India for the period of 2006 to 2010, and observed that the FIIs investment has shown significant improvement in the liquidity of stock prices of both BSE and NSE. Study argued that there is a high degree of positive co-efficient of correlation between FIIs investment and market capitalisation which reveals that the liquidity and volatility in indices are highly influenced by FIIs flows. Dr. Ambuj Gupta (2011)13 examined the relationship between Indian Stock Market and FIIs investment in India for the period from April 2006 to February, 2011 and proved through tests that there is a positive relationship between stock market and FIIs investment which means when FIIs purchase/sell, there is an influence on the stock market. Consequently, either the stock market rise or fall on account of FIIs activities.
4 Statement of Problem
The growing participation of FIIs in Indian stock market has raised eyebrows of many Indians. Their influence on stock markets in India has been widely debated and remained a hot topic in media. Some of the market pundits believe that FIIs are responsible for rise or fall in the Indian stock market. This raises a question as to whether FIIs are really a cause or effect of the rise or fall in the Indian stock market. In this paper the Researchers make an earnest attempt to study the relationship between FII?s Investment and Indian Stock Market. For the purpose, two major stock indices viz. NSE and BSE have been selected. There are certain other significant factors also, which influence Stock market like inflation, government policies, budgets, economic factors etc. However, in this paper only one independent variable i.e. FII?s investment has been taken.
5 Research Gap
There are contradictory findings by various researchers regarding the causal relationship between FIIs investment and stock market movement in India. Therefore, there is a need to investigate whether FIIs are the cause or effect on stock market volatility in India.
6 Scope of the Study
The present study tests impact of FII?s investment on Indian Stock Market. With India emerging to be one of the leading destinations for Foreign Investments, the role and importance of FII?s in the last few years has increased manifold as a result of globalization of the markets. This study covers the period of 12 years i.e.; from 2000 to 2011. The FII?s are emerging to be a key driver in the movement of stock index and it is imperative to study the relation in order to develop an understanding about the efficiency of the market. In the last decade FII had a significant impact on the unprecedented growth in the Sensex and also in its downfall due to the financial recession of 2007. An attempt is made to carve out a clear picture of the impact of FII?s investment on Indian bourses and also determine the market trend relating to inflows and outflows of FIIs.
7 Objectives
1. To assess the growth and development of Indian Stock Market
2. To develop an understanding about the concept and role of Foreign Institutional Investors (FIIs) in India.
3. To study the relationship between FII?s investment and Indian Stock Market
4. To test the impact of FIIs investment on Indian Capital Market
8 Hypothesis
A. Testing Index Volatility of BSE Sensex with FIIs investment
H0: FII's investment has significant relation with Volatility of BSE Sensex
H1: FII's investment has no significant relation with Volatility of BSE Sensex
B. Testing Index Volatility of SandP CNX Nifty with FIIs investment H0: FII's investment has significant relation with Volatility of SandP CNX Nifty H1: FII's investment has no significant relation with Volatility of SandP CNX Nifty
C. Testing the impact of FIIs investment on Indian Capital Market H0: FIIs investment has significant impact on the Indian Capital Market H1: FIIs investment has no significant impact on the Indian Capital Market
9 Research Methodology
9.1 Data Collection Method
The data analyzed in this paper has been collected from the reliable source i.e. from the Handbook of Statistics and Bulletin published by the Securities Exchange Board of India (SEBI) and Reserve Bank of India (RBI), Indian Securities Market Review, NSE fact book from 2000 to 2010 and internet. The sample consists of yearly net changes in two major stock indices of India viz. BSE Sensex and SandP CNX Nifty, and yearly FII?s Net investment in India as on January, 2000 to December, 2011. The data collected is compiled in the form of tables and graphs and scrutinized through statistical tools and techniques.
9.2 Hypothesis Testing Technique
In this paper, researchers test index volatility of Indian stock market. And for this two major stock indices have been taken i.e. BSE Sensex and SandP CNX Nifty. There may be other factors also on which stock market may depend like inflation, government policies, budgets, FDI, economic and political conditions etc. But in this study only one variable i.e. FII have been selected in order to study its relation with stock market volatility. In this paper the concept of correlation and regression is being. Moreover, ttest is being employed here (as the numbers of observations are less than 30).
10 Analysis and Result
10.1 Analysis and Result based on Graph
The Charts or Graphical analysis are based on Table-IV. Charts are the best medium through which the study shows the trends of the market. Here with the help of the graphs the study has analyzed the comparative trend of Net FII?s investment in equity with net changes in two major stock indices of Indian Stock Market i.e. Sensex and SandP CNX Nifty and the market?s prime movers spent to move each point in Sensex and Nifty. These graphs also show the relationship between all the three variables i.e. NSE, BSE and FII. Through this study can analyze the impact of FII?s investment on Indian Stock Market as well as Indian Capital Market.
The figures in Table - IV have been depicted in the Figure 4, 5 and 6. From the Figure 4, 5 and 6 study can analyze the comparative trend of FII?s net investment in equity and net change in Indian Stock Market indices for the period 2000- 2011. A business Standard Research Bureau study of monthly inflows data since 2001 shows the market?s prime movers have spent between Rs 5 crore to Rs 50 crore to move each point in Sensex.14 The charts shows that there is an increase in net investment in 2002-05 due to which Sensex and Nifty also rises, then there was a sharp fall in the year 2006. After that there was a steep increase in net investment in the year 2007. This was the best period in Indian Stock Market where stock prices were at a record high and the market was bullish. Foreign investors have been net buyers in 10 of 12 calendars years since 2000. They have been net sellers in 2008, when the sub-prime crisis and Lehman brothers collapse crippled the global markets. That year they spent Rs 5 crore and Rs 16.78 crore for each of the 10667.96 points the Sensex and 3177.60 points the Nifty lost. And in calendar year 2011, Delivery-based buying in the secondary market has hit a fouryear low, down 25 per cent over the past 12 months, as investors preferred to stay away from a gloomy equity market. So, too, for risk appetite, with average trading volume on the BSE and NSE down 23 per cent over the level a year before. The market was volatile and the reduction in the FII?s investment was one of the causes of volatility. FIIs has pulled out around Rs 53,309.7 crore and Rs 3417.70 crore from the Indian stock market and the withdrawal led to a fall by approximately 51 percent and 24.64 percent in Sensex and 20 percent and 24.62 percent in Nifty in 2008 and 2011 respectively. The post-Lehman recovery that started in April 2009 and got an upward push with return of the UPA government at the center, was even more expensive, at Rs 22.7 crore per point in the Sensex.14 In terms of cost per point in Sensex and Nifty, 2004 was the most expensive for FII, when they spent Rs. 38.05 crore and Rs. 138.83 crore for every point in Sensex and Nifty respectively. In year, 2010, was next on the list, with every Sensex and Nifty point costing Rs. 34.94 crore and Rs. 113.60 crore respectively when in last year, 2011 again it bounce backed Rs. 0.68 crore and Rs. 2.26 crore for cost per point in Sensex and Nifty respectively. 2011 was one of the worst years for Indian markets as it fell more than 20% on a Year on Year basis, which makes it the worst performer of 2011 in Asia. Slowing growth, rising inflation, and policy paralysis have blown the Indian markets off course in 2011. 2011 saw high inflation rate dampening the market spirit.
Headline inflation, as measured by wholesale price index (WPI), has been above the 9% mark since December 2010. This in turn led the RBI action of hiking the repo and reverse repo rates consistently 11 times. And now the Interest rate cycle in India is at the verge of peaking out. However, even if this action by RBI could not tame inflation immediately it helped in reducing the growth story of India. The hike in interest rate, inflation and lack of demand in global markets put huge pressure on Indian markets. The IIP data too saw some shameful numbers with October 2011 IIP registering a negative growth of 5.1%. The GDP growth of India was revised down to 7.6 % for the FY 12 by RBI. Indian Rupee too witnessed huge depreciation in value this year. USD/INR crossed 54 levels in December 2011. RBI's move to bring down speculation in the FOREX market has imparted some stability to the rupee. But the move has negatively affected the foreign inflows. The Great Indian Dream seems to be coming to a close as most of the FIIs are pulling out of Indian markets. The FIIs were net sellers this year pulling out their positions.
Figure 6 shows the relationship of FII?s investment with the BSE Sensex and SandP CNX Nifty. From this it can be seen very clearly that the peaks and the troughs of FII coincide with the peaks and troughs of Sensex and SandP CNX Nifty. From all this it can be analyzed that the FII influence the Indian Stock as well as Indian capital market. The graphical analysis indicates the relation is positively correlated and all three appear to be moving in tandem. The rise and peak of all the three curves appear closely related. While the curves for BSE and Nifty are overlapping each other that of FII investments seems to be following the same pattern or trend. It highlights the significant role and proportion of FII on the movement of stock market and also raises certain questions on the basis of past performance. The role is so prominent that withdrawal leads to a massive fall and increase in investment leads to a corresponding rise. Hence, our markets seemed to be reliant and dependent on FII?s raising questions about stability and reliability from the point of view of domestic investors.
10.2 Analysis and Result based on Hypothesis
This study employs the technique of correlation and regression between FII and Sensex and FII and SandP CNX Nifty in order to test the index volatility of Indian stock market in the context of FII?s investment. First of all, the relationship between these three variables viz; NSE, BSE and FII should be analyzed. In the present study, data has been taken on annual basis and it may be so that the data on monthly or daily basis may provide different result.
10.3 Correlation, Regression and T-Test Analysis
Correlation and regression is being calculated in this study in order to analyze the result and t-test is being employed here in order to test the statistical significance of the results calculated which is depicted in Table V, VI and VII. In Table- V Karl-Pearsons? Product Moment Correlation is being calculated which is a simple correlation and shows the relationship between one dependent variable and one independent variable. Here, FII is taken as an independent variable and Sensex and SandP CNX Nifty are being taken as dependent variables, which are taken one by one for the purpose of calculation.
The above table indicates that the correlation between Sensex and FII?s investment is 0.839 which shows high degree of positive correlation. The R-square is 0.704 which means 70.4% of the variance in the variables is explained by this relationship. According to t-test the calculated value at 5% significance level if 0.053 which lies between the critical values + 2.20 reveals that study is not able to reject the Hull Hypothesis. Hence there is significant relation between FII and Sensex. The above table also shows the correlation of coefficient 0.844 of Nifty with FII which indicates there is strong positive correlation. The R-square is 0.712 which implies that 71.2% of the variance in the variables is explained by this relationship. The calculated t-value is 0.048 which lies within the critical values + 2.20 leads not able to reject the Null Hypothesis. Hence there is relation between Nifty and FII. It is further observed from above table that correlation coefficient between BSE Sensex and SandP CNX Nifty is 0.997 that is a very strong positive correlation. And R-square is 0.995 which reveals 99.5% of the variance in the variable is examined. The calculated t value is 0.712 which lies between critical values + 2.16 which empirically proved that the study can not able to accept the Alternative Hypothesis. Hence there is a significant relation of Sensex with Nifty.
Table VI highlights that there is a linear relationship between the variables. It is observed that the value of the slope is 0.098 signifying that for every unit change in X that stands for FII there is a 0.098 unit change in Y or Sensex. On the other hand the intercept is at -1907.21 which seem that FII is playing hottest role in the volatility of Sensex. There are numbers of indicators which influence in the index volatility of Sensex but FII is one of most influencing factor towards index volatility. Significance value is calculated as 0.000649, which is less than the critical value of 0.05 which means not able to accept the Alternative Hypothesis. Hence there is an impact of the FII on the movement of the Sensex.
Table VII highlights that there is linear relationship between the variables analyzed. It is observed that the value of slope is 0.030, which reveals that for every unit change in FII the value of Nifty is moved by 0.030. On the other hand the intercept at -561.31 indicating the role of FII in the movement of Nifty. It means that if the value of FII is zero then the value of NSE or Nifty would be affected by -561.31 units. The significance value is calculated at 0.000555, which is less than the critical value of 0.05. Again it leads to the acceptance of Null Hypothesis. Hence there is an impact of FII on the volatility of Nifty Index. The both the tables VI and VII support fail to reject the Null Hypothesis which signifies that there is strong relation of FII with volatility of BSE Sensex and SandP CNX Nifty, the effect is slightly more pronounced Nifty. It leads to conclusion that FII?s investment has significant impact on Indian Capital Market.
1. Conclusion
Through the statistical tests, data analysis and graphical presentation it has been found that there is significant relation between the FII?s net investment in equity and volatility in BSE Sensex and SandP CNX Nifty as well as between two indices BSE Sensex and SandP CNX Nifty. Hence it is referred that every movement of FII?s investment there is an instant reaction in the Indian Capital Market. The other factors are also influencing on movement of the stock exchanges. The macro factors in the form of the change in the interest rate, inflation and demand and supply in the global market which Indian market has faced in year 2011. Moreover the sovereign debt crisis in Europe took the center stage in pulling the global market down. The factor might be micro in terms to relating profitability and operations of the listed companies or the economic health of the nation. While exploring the impact on volatility in the stock exchanges it should be kept in mind that FII flows are major drivers of stock markets in India and hence a sudden reversal of flows may harm the stability of its markets it was seen in financial recession in year 2007 and 2011 where the FII is the net buyer from 2000 to 2011 excluding two years 2008 and 2011 which leads the heavy investment and selling attitude of FII?s causing a major hurdle in stabilization of market sentiments.
2. Suggestions
The investment scenario in India has been witnessing turbulent times due to high inflation, poor policy implementation by the government. The problems were further accentuated by the Euro zone crisis and pull backed investment by the FIIs leading to a more than 24% decline in Indian Equities in 2011. Strong policy decisions from the government?s side will make India an attractive investment destination which may attract more FII investments for growth of Indian stock market and making the rupee stronger. There is no doubt FIIs are influencing the volatility on indices of stock exchanges to a greater extent. But the role and effect of the FII on the Indian economy should be duly monitored and regulated by government agencies as our economy is still in the developing stage. It is imperative to ensure that the domestic investors are protected from the established foreign players
Englishhttp://ijcrr.com/abstract.php?article_id=2300http://ijcrr.com/article_html.php?did=2300[1] Link Model, developed by Narendra Singh Bohra available in “FIIs Investment in Indian Capital Market: A Study of Last one Decade”, International Research Journal of Finance and Economics, Issue 68 (2011), pp .103
[2] Purnendra Verma (2001), “Impact of FII on Capital Market – An Empirical study on Indian Capital Market”, pp. 1-19, retrieved from www.scribd.comdoc6988166Impactof-Fii-On (07/12/2011)
[3] Aravind, T. N, et al. (2008) “FII?s Influence in Indian Stock Market”, retrieved from http://www.coolavenues.com/know/fin/subystock-1.php (10/12/2011)
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[9] A.Q.Khan and Sana Ikram (2010), “Testing Semi-Strong Form of Efficient Market Hypothesis in Relation to the Impact of FIIs Investments in Indian Capital Market”, International Journal of Trade, Economics and Finance, Vol. 1, No. 4, pp. 373-379
[10] Narendra Singh Bohra and Akash Dutt (2011), “FIIs Investment in Indian Capital Market: A Study of Last one Decade”, International Research Journal of Finance and Economics, Issue 68, pp .103-116
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[15] http://www.sebi.gov.in
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[19] http://www.moneycontrol.com
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[21] http://www.google.com [22] SEBI handbook of statistics on the Indian Securities Market 2009, 2010
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[26] NSE factbook from 2000 to 2011
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareSTUDY OF LIPID PEROXIDATION, ANTIOXIDANT STATUS AND LIPID PROFILE IN BREAST CANCER
English4751G.S.R.KedariEnglish G.S.R.HareeshEnglish A. SaseekalaEnglishOBJECTIVE: Breast cancer is the commonest malignancy in women in India and western countries. Increased production of oxygen free radicals exhaust the antioxidant levels in the body leading to the development of oxidative stress which results in the production of cancer. The reasons for alteration of lipid profile in breast cancer is not clearly understood. The aim of our present study is to evaluate the role of oxidative stress and also the status of serum lipid profile in breast cancer patients. METHODS: The role of oxidative stress in breast cancer patients were evaluated by estimating the levels of lipid peroxidation assessing plasma Malondialdehyde(MDA) levels and antioxidant status by reduced glutathione(GSH), Vitamin-C in blood. We also tried to assess lipid profile by estimating Serum total cholesterol, triglycerides, HDL cholesterol and LDL cholesterol. For this, we have taken 50 cases of breast cancer patients compared with 50 age matched control women. RESULTS: There were significant increase in the levels of MDA and significant decreases in the levels of antioxidants like GSH and Vitamin-C in breast cancer patients when compared with controls. There were also significant increase in the levels of Serum total cholesterol, triglycerides and LDL cholesterol and no statistical significant difference was observed with HDL cholesterol in cases when compared with controls. CONCLUSION: Our results indicate that oxidative stress and alterations in lipid profile are associated with the development of breast cancer and the need for modification of lifestyle for the reduction of breast cancer development.
EnglishOxidative stress, Malondialdehyde, Reduced glutathione (GSH) and Vitamin-C.INTRODUCTION
Breast cancer is one of the most common neoplasm?s in women and is a leading cause of cancer related deaths worldwide. The aetiology of breast cancer is multifactorial. Epidemiologic studies have identified many risk factors that increase the chance of a woman developing breast cancer include early age at menarche, late age of menopause, null parity, obesity, oral contraception, hormone replacement therapy, diet, family history, prior history of benign breast disease and lactation. The common denominator for many of these risk factors is their effect on the level and duration of exposure to endogenous or exogenous estrogens.
Oxidative stress is implicated in the pathogenesis of a variety of human diseases(1). Oxidative damage occurs to biomolecules like lipids, proteins, carbohydrates and nucleic acids and other extracellular components like collagen and hyaluronic acid which are very deleterious(2 )resulting in lipid peroxidation, mutagenesis and carcinogenesis. However, the body?s defense mechanisms play an important role in the form of antioxidants that help to minimize the damages which are caused by oxidative stress. Antioxidants are compounds that dispose, scavenge and suppress the formation of free radicals or oppose their actions. Oxidative stress occurs when there is an imbalance between reactive oxygen species(ROS) and antioxidants reaction capacity which stimulate the development of a disease such as breast cancer(3).Several case control studies have reported a relationship with antioxidant status(4,5) and a reduction in antioxidant level due to the presence of free radicals may increase risk of breast cancer. The neoplastic disease is related to new growth where there is greater utilization of lipids including total cholesterol, lipoproteins, and triglycerides for new membrane biogenesis. Cells fulfill these requirements either from circulation, by synthesis through the metabolism or from degradation of major lipoprotein fractions like VLDL, LDL or HDL(6). The aim of our study was to evaluate the role of lipid peroxidation in breast cancer by estimating the plasma MDA levels and the role of enzymatic and non enzymatic antioxidants by estimating reduced glutathione and vitamin-C and also to evaluate the relationship between the lipid profile and breast cancer.
MATERIAL AND METHODS
The present study was conducted in the department of biochemistry and department of general surgery in Saveetha medical college and S.V. Medical college, Tirupati.50 newly diagnosed cases of breast cancer belonging to age group of 30-70 years were included in this study. Out of cases,43 were having ductal carcinoma and 6 patients were having lobular carcinoma and 1 was having mixed carcinoma(both ductal and lobular).75 age matched women who have no history of breast diseases were taken as controls. All the subjects were not using any kind of hormonal therapy and they had no any other diseases like Diabetes mellitus, liver diseases and thyroid disorders. Informed consent was obtained from all the subjects. Due permission was obtained from Ethical clearance committee for this study. 10ml of fasting blood samples were collected by venipuncture and for the separation of sera, 5ml of blood was centrifuged at 3000rpm for 5min and the remaining 5ml of blood was taken into a plain vial containing EDTA and was centrifuged at 3000rpm for 10min for the separation of plasma. The plasma MDA levels were estimated by using thiobarbituric acid reacting substances(TBARS) by the method of Yagi(7) and Sinnhuber et al(8).Reduced glutathione was determined by the method of Beutler et al(9). The activity of Ascorbic acid was determined by the method of Tietz(10).Serum was used for the estimation of lipid profile. Total cholesterol and triglycerides were estimated by enzymatic methods(11,12).HDL cholesterol(HDL-C) was estimated by phosphotungstic acid precipitation followed by enzymatic analysis in supernatant fraction(13) and LDL-cholesterol was determined by using Friedwald?s equation(14). All the results were expressed as mean ± SD and statistical comparisons were done using student t-test using the SPSS package.
Evaluation of oxidative stress is done based on the levels of MDA and statistically significant increase in the level of MDA was observed in breast cancer patients when compared to controls. Statistically significant decreases were observed in the levels of antioxidants like GSH and Vitamin-C in cases when compared to controls. There were also significant increases in the levels of serum total cholesterol, triglycerides and LDL cholesterol in cases when compared to controls. There was no statistical significant difference in the levels of HDL in cases when compared with controls.
DISCUSSION
Damage to the breast epithelium by oxygen free radicals can lead to fibroblast proliferation, epithelial hyperplasia, cellular atypia and breast cancer. Studies have shown increased lipid peroxidation in solid tumors(15,16).The significant rise in the MDA levels in breast cancer confirms that it is associated with an increased production of reactive oxygen species(ROS) and free radicals. Lipid peroxidation is a chain reaction which provides a continuous supply of free radicals that initiate further peroxidation(17). We also observed a significant decrease in the levels of reduced glutathione in the cases as compared to the controls. The intracellular depletion of reduced glutathione can be either due to the formation of a direct complex with a electrophilic agent or due to the inhibition of synthesis or due to the subjection of the cell to oxidative stress(18). When a cell is subjected to oxidative stress, there is increased utilization of glutathione, thus leading to its depletion. Many enzymes are GSH dependent and their activity may be regulated by the thiol disulphide exchange. They are thus dependent on the GSH status. Glutathione –S- transferase (GST) is reduced in diabetics which are dimeric, mainly cytosolic enzymes that have extensive ligand binding properties in addition to their catalytic role in detoxification(19,20). This reduction is due to the reduced levels of GSH. There is also a decrease in the levels of non-enzymatic anti-oxidants such as Vit-C, which states that there is an increased defense mechanism against oxidative damage in breast cancer. The decrease in the levels of these non-enzymatic antioxidant parameters may be due to an increased turnover for preventing oxidative damage in these patients, thus suggesting an increased defense against oxidative damage. Our results support the researchers who reported decreases in the antioxidant level and increases in lipid peroxidation level(21,22).Several other researchers showed over expression of antioxidants(16,23). In the present study, there is significant increase in the levels of serum total cholesterol, LDL cholesterol and triglycerides in the breast cancer patients when compared with normal subjects and there is no significant difference in the levels of HDL cholesterol . The patho physiological mechanism for lipid alterations underlying is not well understood. Lipids are major cell membrane components essential for various biological functions including cell growth and division of normal and malignant tissues. Low levels of cholesterol in the proliferating tissues and in blood compartments could be due to the process of carcinogenesis. The raised plasma concentrations of theses parameters in patients with breast cancer may be due to an increased rate of lipid absorption as the fat splitting enzymes, lipases, were also found to be increased in the patients(24). In conclusion, the present study demonstrated high oxidative stress, low antioxidant status with rise in plasma lipid levels except HDL in breast cancer patients which shows the need to adopt a healthy lifestyle to reduce oxidative stress with consumption of diet rich in antioxidant nutrients to prevent breast cancer.
CONCLUSION
Our results indicate that oxidative stress and alterations in lipid profile are associated with the development of breast cancer and the need for modification of lifestyle for the reduction of breast cancer development.
ACKNOWLEDGEMENTS
Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2301http://ijcrr.com/article_html.php?did=23011.Beck MM, Levander OA. Dietary oxidative stress and potentiation of viral infection. Annu Rev Nutr,1998; 18: 93-116.
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3.Aghvami T, Djalali M, Kesharvarz A, et al. Plasma level of antioxidant vitamins and lipid peroxidation in breast cancer patients. Iran J Publ Health,2006; 35: 42-7.
4.Ching S, Ingra D, Hahnel R, Beilby J, Rossie E. Serum levels of micronutrients, antioxidants and total antioxidant status predict risk of breast cancer in a case control study. J Nutr, 2002; 132: 303-6.
5.Do MH, Lee SS, Jung PJ, Lee MH. Intake of dietary fat and vitamin in relation to breast cancer risk in Korean women: a case control study. J Korean Med Sci,2003; 18: 534-40.
6.M.I.Qadir, S.A.Malik. Plasma lipid profile in gynecologic cancers. Eur.J.Gynaec.oncol, 2008; 29: 158-161.
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17. Murray RK, Granner DK, Rodwell VW. Harper?s Illustrated Biochemistry. 27th Edn.2006.Mc Graw Hill Lange International Edition.pp128-129.
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21. Gonenc A, Erten D, Aslan S, et al. Lipid peroxidation andantioxidant status in blood and tissue of malignant breast tumor and benign breast disease. Cell Biol Int 2006;30:376-80.
22. Yeh CC, Hou MF, Tsai SM, et al. Superoxide anion radical, lipid peroxides and antioxidant status in the blood of patients with breast cancer. Clin Chim Acta 2005;361:104- 11.
23. Iscan M, Coban T, Cok I, et al. The organochlorine pesticide residues and antioxidant enzyme activities in human breast tumors: is there any association? Breast Cancer Res Treat, 2002;72:173.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcarePSYCHOLOGICAL STRESS AND ORAL DISEASES - A LINK ?
English5258Little MahendraEnglish Ravi David AustinEnglish Jaideep MahendraEnglish S.Senthil KumarEnglishEnglishINTRODUCTION
Epidemiologic studies and clinical observations suggest that negative life events and psychological factors may contribute to an increased susceptibility to periodontal disease. Stress a term continually being redefined in the scientific study of disease and illness, is nevertheless a confirmed and important factor in the etiology and maintenance of many inflammatory diseases, including periodontal disease. stress is an attempt to understand how the body regulates itself to maintain smooth, adaptive and homeostatic functioning when confronted with disruptive endogenous or exogenous forces. Hippocrates thought of health as a harmonious balance of the elements comprising the quality of life while disease represented disruption or disharmony among those elements. In the seventeenth century, Sydenharm suggested that pathological states represented diseases of adaptation failure of the adaptive processes to restore wellbeing. Cannon elaborated how fight or flight mechanisms represented adaptive efforts by the body to reestablish homeostasis, a term he introduced to describe a dynamic internal physiological equilibrium which the body sought to maintain along both physical and emotional dimensions. Hans Selye, the father of stress theory, describes stress as, “Everybody knows what it is, no one knows what it is. It is the nonspecific response of the body to any demand put upon it and stress is the spice of life” (1). While Stress is defined as a state induced by a stimulus that manifests itself by virtue of one?s cognitive interpretation (2), the stimulus itself is considered a stressor. Accordingly, a stressor is any stimulus that evokes stress, whereas stress reactions are the observable consequences of the stressors. Several studies have demonstrated the relationship between psychological stress and diseases, from common cold (3) to cardiovascular disorders (4); from asthma (5) to rheumatoid arthritis (6). Periodontal disease being multifactoral with a complex interaction between bacterial infection and host responses, there is a reasonable amount of research indicating the association between periodontitis with psychosocial stress, distress, and depression (7, 8). When demands imposed by events exceed a person?s ability to cope, a psychological stress response composed of negative cognitive and emotional state is elicited (9). Substantial literature both in humans and animals support association between psychological stress and suppression of immune responses (10). While stress can directly influence immune function through the activation of neuro-endocrinal pathways that lead to release of hormones and neurotransmitters, such as cortisol and catecholamines (11); it can also alter immune responses through the adoption of coping behaviors, such as smoking or drinking alcohol, which are known to compromise immunity (12).
Pathways between Stress and the Immune System:
How can stress affect the immune response? In 1936, Hans Selye defined stress physiologically as the state in which the sympatho-adreno-medullary system and the limbic-hypothalamic-pituitary-adrenal axis (HPA) are co-activated (13). A bidirectional communication pathway exits between the CNS and the immune system that modulates both the cellular and humoral immunity (14). First, stress induces the sympathetic fibers which descend from the brain into both primary (bone marrow and thymus) and secondary (spleen and lymph nodes) lymphoid tissues (15) to release a wide variety of substances that influence immune responses by binding to receptors on white blood cells (14, 15). Though all lymphocytes have adrenergic receptors, differential density and sensitivity of adrenergic receptors on lymphocytes may affect responsiveness to stress among cell subsets. For example, natural killer cells have both high-density and high affinity β2-adrenergic receptors, B cells have high density but lower affinity, and T cells have the lowest density (16, 17).Second, the stress activates the hypothalamic–pituitary–adrenal axis, the sympathetic–adrenal–medullary axis or locus coeruleus-norepinephrine system, to induce secretion of adrenal hormones epinephrine, norepinephrine, and cortisol (13, 15); and also the pituitary hormones prolactin and growth hormone; and the brain peptides melatonin, β-endorphin, and enkephalin. These substances bind to specific receptors on white blood cells and have diverse regulatory effects on their distribution and function (18).Communication between the neuroendocrine (hypothalamic-pituitary-adrenal) and immune inflammatory systems functions as a feedback loop that regulates the immune components of the inflammatory response. For example, a negative feed back loop functions such that activation of the immune system, associated with increases in levels of circulating cytokines (e.g. interleukin-1 and interleukin-6), increases activity in the corticotrophin releasing hormone/hypothalamic-pituitary-adrenal system yielding increased levels of circulating adrenocorticotropic hormone and cortisol – major modulators of the stress system (19).
Connection between Stress and Periodontitis:
Stress can be viewed as a process with both psychological and physiological components. Pollman and Dietrich (1979), Moulton et al (1952) pointed out that stress may affect periodontium directly or indirectly (7). While the psychosocial stressors initiate cascade of events through the hypothalamic-pituitary-adrenal axis, the autonomic nervous system and the central nervous system, it enhances the likelihood of infection and specifically, periodontal disease (8, 20).While cortisol has been called the hormone of stress, serum cortisol level increases during challenging or unpleasant situation (21, 22). Cortisol being an immunosuppressant influences not only the inflammatory cells, chemotaxis etc., but also the pro-inflammatory cytokines like interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1ra) (23, 24, 25). It is always being studied that stress causes activation of neuro-endocrinal pathways with release of cortisol, which in turn suppresses the immunity. And in such an environment the progression of periodontal infection by the pathogen is unhampered (7). Though this relation gives a link between stress and periodontal disease, this holds good for other inflammatory diseases as well. Though the periodontitis is initiated by the pathogens, the mediators of connective tissue breakdown are produced by hostderived enzymes called Matrix metalloproteinases (MMPs) (26). They are zinc and calcium dependent proteolytic enzymes responsible for remodeling and degradation of extracellular matrix (27). The homeostasis of extracellular matrices depends on the release of MMPs from cells such as fibroblasts and macrophages, and the presence of tissue inhibitors of matrix metalloproteinases (TIMPs), which are distributed widely in tissues and fluids (28). While the MMP gene family encodes nine or more metal-dependent endopeptidases, eight human MMPs have been cloned and sequenced, of which MMP-1 cleaves fibrillar collagen types I, II, and III (27) that constitute the gingival and periodontal connective tissue. While MMP-1 is implicated to play an important role in the initiation of collagen degradation in periodontal disease, MMP-8 is thought to play an important role in periodontal tissue destruction (29). Very few in-vitro studies, using exogenous steroids, have shown the effects of corticosteroids on the expression of MMPs and TIMPs in fibroblasts (30, 31). A study by Cury PR et al. (31) showed that hydrocortisone produced a dose-dependent regulation of MMP and TIMP expression, by significantly up-regulating mRNA expression of MMP-1, -2, -7, and -11 and TIMP-1 in human gingival fibroblasts. When Pubmed was searched for studies where endogenous cortisol, due to stress, induced expression of MMPs and TIMPs in fibroblasts, none were found.
Significant Reviews:
Several studies have demonstrated a relationship between psychological stress and inflammatory diseases such as rheumatoid arthritis, osteoarthritis etc., (20, 32, 33) and periodontitis (7, 8).Gerard J. Linden et al. (34) examined the association between occupational stress and the progression of periodontitis in employed adults and suggested that occupational stress may have a relationship to the progression of periodontitis. Torbjørn Breivik et al. (35) reviewed studies to find effects of emotional stress on immunity, gingivitis and periodontitis, and noted that emotional stressors and the nervous and neuroendocrine responses to psychological stressors may modulate the immune response to bacteria, and thus expected stress to influence the progression and course of gingivitis and periodontitis.In a case-control study, Moss et al. (36) explored the association between social factors and adult periodontitis by comparing selfreported information for daily strains and symptoms of depression. They found that an elevated Depression score may be a marker for social isolation, which could play a role in immune function during periods of social strain. Croucher R et al. (37) also in a casecontrol study investigated the role of lifeevents in periodontitis. The results showed that periodontitis was associated with the negative impact of life-events, the number of negative life-events, high levels of dental plaque, tobacco smoking and unemployment. Genco RJ, Ho AW, Jeffrey Kopman et al. (38), Genco RJ, Ho AW, Grossi SG et al. (8) evaluated the association of stress, distress, and coping behaviors with periodontal disease. They found that subjects with financial strain and inadequate coping had greater attachment and alveolar bone loss, in contrast to subjects with financial strain and good coping. Salivary cortisol levels were higher in a test group exhibiting severe periodontitis, as compared to a control group consisting of those with little or no periodontal disease. Renate Deinzer et al. (39, 40) analyzed the effects of academic stress on periodontal health of medical students and found that psychological stress was a significant risk factor for periodontal inflammation. Elter et al. (41) found that clinical depression may have a negative effect on periodontal treatment outcome in health maintenance organization (HMO) patients. Study by Hugoson A, Ljungquist B, Breivik T (42) revealed that, in addition to increased age, oral hygiene status, and smoking, the negative life events like, loss of a spouse (being a widow or widower) and the personality trait of exercising extreme external control were also associated with severe periodontal disease. Aleksejuniené t al. (43) tested the hypothesis that psychosocial stress and lifestyle are related to periodontal status. Although the pathway between psychosocial stress and remaining periodontal support was not empirically supported, the researchers concluded that there was reason to believe that such link was likely. Wimmer G et al. (44) examined the influence of different coping behaviors on a non-surgical periodontal therapy and on the course of periodontal disease. They found patients with a defensive coping style had statistically significant poorer attachment values (P= 0.000) after 2 years compared to patients with other coping behaviors. The number of sites with severe advanced CAL (>5mm) was significantly correlated with a suppressive coping style (P=0.0001).Alexander Saletu et al. (45) investigated the relationship between periodontitis and psychopathology utilizing psychometry, both observer-rating scales and self-rating scales. Partial correlation analyses between psychometric measures and dental variables revealed positive correlations of periodontal disease severity/CAL with the depression/anxiety, subjective well-being and complaints scores, and a negative correlation with quality of life. R. Akhter et al. (46) designed a study to identify possible relationship between stress and periodontal disease in residents of a rural Japan and found subjects who felt job stress and those who felt stress due to self health were more prone to have periodontal disease than were those who never or only rarely felt such stress. Vettore et al. (47) in a case-control study investigated the relationship of stress and anxiety with periodontal clinical characteristics. They found the frequency of moderate clinical attachment loss (4-6 mm) and moderate probing pocket depth (4-6 mm) significantly associated with higher trait anxiety scores, after adjusting for socioeconomic data and cigarette consumption. J.B. Hilgert, F.N. Hugo D.R. Bandeira and M.C. Bozzetti (48) evaluated the extent and severity of chronic periodontitis and its association with the levels of salivary cortisol and the scores obtained with a stress questionnaire in a population aged 50 years. They found that the cortisol levels were positively associated with the extent and severity of Periodontitis. Fernando N. Hugo, Juliana B. Hilgert et al (49) evaluated the effects of stress, depression, and cortisol levels in dental plaque accumulation and gingivitis in a population of individuals aged ≥50 years. They found that stress was a significant risk indicator of elevated levels of plaque and gingivitis, whereas cortisol was a risk indicator of plaque. Amy E Rosania (50) did a cross-sectional pilot study to explore the associations between psychological factors, markers of periodontal disease, psychoneuro-immunologic variables, and behavior. She found that stress, depression, and salivary cortisol correlated with measures of periodontal disease. In addition, oral care neglect during periods of stress and depression was associated with loss of attachment and missing teeth.
CONCLUSION
Stress can be a risk factor for periodontitis, on one hand in stress the person?s oral hygiene habits are altered causing accumulation of plaque and on the other it reduces the immunity of person through its endocrinal connections. Many studies have shown a positive relationship was observed between stress and periodontal disease, further representative research is needed to determine the impact of stress/psychological factors as risk factors for periodontal disease.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareAN IN VITRO SCREENING OF GROWTH INHIBITORY POTENTIAL OF ALLIUM SATIVUM TOWARDS SOME MICROBES OF SPOILAGE AND HEALTH SIGNIFICANCE
English5966Mamta BhatiaEnglish Alka SharmaEnglishThe use of Allium sativum (garlic) as a cure and condiment predates written history. In present study aqueous extract, crude juice, essential oil and powdered form of Allium sativum were screened for their inhibitory potential towards some food borne pathogens, in culture media. Test microbes included : Bacillus cereus, Enterococcus faecalis, Escherichia coli, Psuedomonas aeruginosa, Psuedomonas alkaligenes, Shigella sonnei and Staphylococcus aureus. Spice agar method was opted for investigating
antibacterial activity of powdered spice samples. Agar well assay and broth dilution techniques were followed for determining growth inhibitory potentials of aqueous extract, crude juice and essential oil. Results revealed that essential oil most effectively inhibited bacterial strains followed by crude juice, while aqueous extract and powdered forms remained ineffective in arresting the growth of test bacteria.
EnglishAntibacterial, Antimicrobial, Allium sativum, essential oil, garlic, spicesINTRODUCTION
Foods, by their very nature need to be nutritious and microbiologically stable . To ensure that food is safe and can be stored in a satisfactory state, it is necessary to either destroy the microorganisms present, or manipulate the food so that microbial growth is prevented or hindered. Resurgence in the use of natural herbal alternatives instead of synthetic preservatives to increase the shelf life of food commodoties has brought the use of aromatic plants to the forefront of investigations. Allium sativum, is one of the world?s most popular spices, and is used extensively from India to America, in French aioli, Greek skordalia, Indian korma, Turkish cacik and Vietnamese pho bo. Since ancient times, it has been used as a cure as well as food. Pliny the elder, a Roman naturalist, described in his Historia Naturalis how A. sativum could be used for gastrointestinal disorders, dog and snake bites, scorpion stings, asthama, madness, convulsions and tumours. Components from A. sativum modulate the cardiovascular and immune systems. Alongwith medicinal properties, it is known to have antiviral1,2 and antiprotozoal3,4 activities. Encouraged by these results, an in vitro trial was carried out to evaluate different forms of A. sativum viz. aqueous extract, crude juice, essential oil and powdered form, for their antimicrobial potencies, against seven food borne pathogens.
Materials and Methods
Procurement of spice samples
Fresh bulbs of A. sativum purchased in the amounts of 1 kg, from grocery shop, local market, Hisar, India. The spice samples were washed with clean water followed by distilled water to remove extraneous matter. The outer coverings of A. sativum bulbs/clove were peeled off manually with the help of knife. For the extraction of crude juice, peeled cloves of A. sativum were sliced into thin pieces and were crushed in pestle-mortar to get a thick paste. Thick pastes of spice samples were passed through sieve cloth. Filtrates thus obtained were sterilized by passing through syringe filter assembly having membrane filters of pore size 0.45 um under aseptic conditions. Crude extract thus obtained was stored in sterilized glass vials at 4+1° C and was used at various concentration levels within the 2 h. of their preparation. To get the powdered form , peeled cloves of A. sativum were dried in shade for 5 days followed by their grinding in the laboratory grinder and were kept in airtight containers till further use. Essential oil of A.sativum was procured from Aroma Chemicals Pvt. Limited, Delhi, India, stored in the dark amber colored, screw capped glass bottle and was kept away from light to avoid physicochemical changes in its composition. Purity of the essential oil was assured by the company to be more than 99.0 % .
Chemicals and culture media
Ethyl violet azide dextrose agar, Ethyl violet azide dextrose broth, MacConkey broth, MacConkey agar, Nutrient agar and Nutrient broth were obtained from HiMedia Pvt. Ltd, India. Dimethylsulphoxide (DMSO) and Sodium chloride (NaCl) were obtained from Central drug house Pvt. Limited, India.
Bacterial cultures
All the pure bacterial cultures viz. Bacillus cereus (MTCC 430), Enterococcus faecalis (MTCC 439), Escherichia coli (MTCC 1687), Psuedomonas aeruginosa (MTCC 1688), Psuedomonas alkaligenes (MTCC 405), Shigella sonnei (MTCC 2957) and Staphylococcus aureus (MTCC 5021) were obtained from Microbial Type Culture Collection (MTCC), Institute of Microbial Technology (IMTECH), Chandigarh, India. The reference bacterial strains were maintained on their respective media slants, subcultured bimonthly to maintain their viability and were stored at 4+1° C. Culture media, incubation temperatures and duration of incubation of reference bacterial strains are presented in Table 1.
Inoculum preparation
A flamed sterile wire loop was used to dislodge the lawns of test bacterial strains from their respective pure culture slants (24h. old) with 10 ml of sterilized normal saline (NaCl, 0.85% (w/v)) solution under aseptic conditions. Bacterial suspensions were adjusted with the same solution to contain approximately 1×107 cfu /ml and were utilized the same day.
Prepearation of aqueous extract
Aqueous extract of dried and powdered bulbs was prepared. Powdered spice sample was steeped overnight (temperature: 24-27°C) in sterilized distilled water in a ratio of 1:1 (w: v), followed by their homogenization in a blender at high speed for 2 min. The homogenized spice mixture was filtered through Whatmann No. 1 filter paper. Filtrate thus obtained, was sterilized by passing through syringe filters containing 0.45 um pore size membrane filters under aseptic conditions, collected in sterilized glass vial and was stored at 4+1° C. This aqueous extract was further used within the 2 h. of its preparation.
Preliminary screening of antibacterial activities of aqueous extract, crude juice and essential oil
Agar-well diffusion technique was followed5 . Freshly prepared inoculum (100 ul) of each reference bacterial strain was poured in plates with 20 ml of appropriate media. The petriplates seeded with bacterial strains were kept undisturbed for 30 min. for proper solidification and setting of agar to facilitate uniform digging of wells. Sterile cork borer (diameter: 8 mm) was used to bore wells in the solidified media plates previously seeded with bacterial inocula. Subsequently, different volumes of test substances were introduced into the wells of agar plates. Sterilized DMSO, instead of crude juice and essential oil served as negative control. These plates were allowed to stand at room temperature for at least 1 h. for the even diffusion of poured components and were incubated without inversion at their respective incubation temperatures in incubator for 24-48 h. After incubation, zones of inhibition formed around the wells were measured in millimeters (mm) and results were expressed as the net zone of inhibition which represented the subtraction of the diameter of the well (8 mm) from the measured zone.
Mnimum inhibitory concentrations (MIC) of crude juice and essential oil
MIC values of crude juice and essential oil were determined by broth dilution method6 . The media (broth) containing 2000 ul/ml of test substance was serially diluted twofold each with the media (broth) to give concentrations of 1000, 500, 250, 125, 62.50, 31.25, 15.62, 7.81, 3.90, 1.95, 0.97, 0.48, 0.24, 0.12, 0.06 ul/ml. Sterilized DMSO, instead of crude juice and essential oil, served as negative control. To the diluted solution, 100 ul of freshly prepared inoculum of each bacterial strain was added. These mixtures were incubated in the B.O.D. incubator at suitable incubation temperatures of microbes, for appropriate incubation periods. After the completion of incubation, 100 ul of the above mixture was evenly spread on the surface of solidified media petriplates with the help of sterile bent glass rod. These petriplates were incubated in an inverted position to observe the minimum concentration of test substances, at which visible growth of the reference microbes was completely inhibited.
Antibacterial activity of powdered form
Antibacterial activity of powdered form of spice sample was examined in culture media using spice agar method7 . Erelenmeyer flasks (100 ml capacity) containing 20 ml of appropriate media (containing agar) and powdered spice at different concentrations (0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0 (%,w/v) were autoclaved at 121° C for 20 minutes. After autoclaving, spice agar mixtures (cooled but still molten) were poured into sterilized petriplates under aseptic conditions and these plates were kept undisturbed for 30 min. for proper setting of agar. Freshly prepared inoculum of each test microbe at 100 ul level was evenly spread over the entire surface of the respective solidified media in petriplate using a sterile bent glass rod. Seeded petriplates were incubated in incubator at appropriate temperatures and were examined for bacterial growth at 12 h. intervals, throughout the incubation period of 30 days. A similar experiment was carried out without any spice sample, that served as control. The time for initiation of microbial growth on control (without spice samples) and media supplemented with different concentrations of spice were recorded.
Statistical analysis :
All the experiments were performed in triplicates with two independent trials and the results obtained were highly reproducible. Values of growth inhibitory zones are mean ±SD (n=3) of three replicates.
RESULTS
Zone inhibition assay results (Table 2) revealed that seeded petriplates with DMSO and aqueous extract of A. sativum , did not display growth inhibitory zones towards any bacterial strain under observation. Crude juice at 100 ul/well level exhibited inhibitory circles towards B. cereus, P.aeruginosa and S. aureus. On the other hand, essential oil of A. sativum, at 10ul/well exhibited distinct zones of inhibition towards all the bacterial strains under investigation. The diameter of inhibitory zones varied with the type of bacterial strains and test substances implicated in the study. Crude juice exhibited widest diameter of inhibitory zone towards S.aureus, while essential oil produced broadest inhibitory circle towards B.cereus. It was also observed that g+ve bacterial strains gave wider inhibitory zones towards test substances as compared to g-ve bacterial strains. On the basis of diameter of growth inhibitory zones, sensitivity of microbes in descending order towards test substances may be put in the following manner: Essential oil: B.cereus>S.aureus>P.aeruginosa>P.alkal igenes>S.sonnei>E.faecalis>E.coli. Crude juice: S.aureus>B.cereus>P.aeruginosa=E.coli =E.faecalis=P.alkaligenes=S.sonnei. Results of broth dilution technique depicted that crude juice and essential oil of A. sativum effctively inhibited all the bacterial strains (Table 3). MIC values of essential oil towards bacterial strains ranged from 62.50-500.00 (ul/ml), whereas, that of crude juice ranged from 125.00-500.00 (ul/ml), thereby indicating the higher susceptibility of the microbes towards former. It was noticed that higher concentrations of crude juice and essential oils were required to inhibit g-ve bacterial strains. Dried and powdered bulbs of A. sativum at all the concentration levels i.e. 0.0, 0.1, 0.2, 0.4, 0.6,0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0 (%, w/v), remained ineffective in arresting bacterial strains, and visible growth of all the microbes was noticed on 2nd day of incubation, as in control set of petriplates, without any spice sample.
DISCUSSION
Functional properties of aromatic plant/spices etc. are contained in their volatile aromatic secrections commonly known as essential oils8 . A. sativum bulbs have 0.1%-0.25% essential oil, which is composed of 60% diallyl disulphide (Allicin), 20% diallyl trisulphide, 6% allyl propyl disulphide and diallyl sulphide9 . The inhibitory activity of essential oil of A. sativum is widely attributed to diallyl disulphide (Allicin). Allicin is highly volatile and is formed by the action of enzyme allinase on alliin (an odourless precursor), when fresh cloves/bulbs of A. sativum are cut or bruised10 . The mode of action of allicin to inhbit growth of bacterial strains is not yet well understood however, it may involve : hydrophobic and hydrogen bonding of active components of essential oil to membrane proteins, perturbation of membrane permeability, leakage of ions and other cell contents, inhibition of membrane embedded enzymes, destruction of electrons transport systems, disruption of proton motive force (PMF) and coagulation of cell contents leading to death. The ineffectivity of aqueous extract and powderd form of A. sativum bulbs towards test microbes in the present experiment may be attributed to the loss of allicin during drying of cloves/bulbs under ambient conditions. The greater susceptibility of g+ve bacterial strains towards crude juice and essential oil of A. sativum may be due to the absence of an outer membrane in their cell membrane which makes them more sensitive to external environmental changes such as temperature, pH , natural extracts, essential oils and other antimicrobial substances. On the other hand, the lipopolysacharides in the cell membrane of g-ve bacteria could provide a barrier to many antimicrobial agents, rendering these bacteria more resistant to certain agents than g+ve bacteria. It is worth mentioning here that crude extract more effectively arrested microbes during broth dilution technique than zone inhibition assay. This may be attributed to the direct contact of the microbes with liquid media which might have allowed the easy and quick diffusion of antimicrobial components of crude juice to the target site.
Conclusion
Present in vitro study indicate that essential oil of A. sativum inhibited food borne pathogens most effectively and may be considered for food preservation. Further studies should be undertaken to elucidate the safety, stability and organoleptic aspects of essential oil and its precise mode of action. Interactions of essential oil components with different food matrices during various food processing treatments must be the focal area of research before their commercialization as „biopreservatives?.
Englishhttp://ijcrr.com/abstract.php?article_id=2303http://ijcrr.com/article_html.php?did=23031. Weber ND, Anderson DO, North JA, Murray BK, Lawson LD, Hughes BG. In vitro virucidal activity of Allium sativum (garlic) extract and compounds. Planta Med 1992; 58 : 417–23.
2. Shoji S, Furuishi K, Yanase R, Miyazaka T, Kino M. Allyl compounds selectively killed human deficiency virus-type 1-infected cells. Biochem Biophys Res Commun 1993;194 : 610–21.
3. Lun ZR, Burri C, Menzinger M, Kaminsky R. Antiparasitic activity of diallyl trisulfide (Dasuansu) on human and animal pathogenic protozoa (Trypanosoma sp., Entamoeba histolyica and Giardia lamblia) in vitro. Ann Soc Belg Med Trop 1994; 74:51–9.
4. Reuter HD, Koch HP, Lawson LD. 1996. Therapeutic effects and applications of garlic and its preparations. In: Koch HP, Lawson LD (eds) Garlic: the science and therapeutic application of Allium sativum L. and related species. Williams and Wilkins, Baltimore, pp 135–213.
5. Iroegbu CU, Nkere. Evaluation of the antibacterial properties of Picralima nitida stembark extracts. International J Mol Med Adv Sci 2005; 1: 182-9.
6. Kim HO, Park SW and Park HD. Inactivation of Escherichia coli 0157:H7 by cinnamic aldehyde purified from Cinnamomum cassia shoot. J Food Micro 2004; 21:105-10.
7. Azzouz MA, Bullerman LB. Comparative antimycotic effects of selected herbs,spices, plant components and commercial antifungal agents. J Food Prot 1982; 45: 1298-1301.
8. Pruthi JS. Spices and Condiments. National Book Trust, New Delhi, India, 1976; pp. 117–21.
9. Stoll V, Seebeck E. Allium compounds. I. Alliin the true mother compound of garlic oil. Helv Chem Acta 1948 ; 31:189.
10. Lawson LD. The composition and chemistry of garlic cloves and processed garlic. In: Koch HP, Lawson LD (eds) Garlic: the science and therapeutic application of Allium sativum L. and related species. Williams and Wilkins, Baltimore, 1996 ; pp 37–107.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareIn-vitro Estimation of Free Radical Scavenging Activity of Fruit juices by DPPH assay
English6773Singh MeenuEnglish Pragzna YanamadalaEnglish Dharmadev BommiEnglishObjective: A number of different beverage products claim to have antioxidant potency due to their perceived high content of polyphenols. The present study was designed to evaluate the in vitro estimation of free radical scavenging activity of three different fruit juices as follows: Grapes, Guava and Pineapple available in local market. Method: Above mentioned fruit juices were screened for their free radical scavenging property using diphenyl picryl hydrazyl (DPPH) radicals. The absorbance of these fruit juices at 517nm is found by using UV-spectroscopy at various time intervals such as 15 minutes, 30 minutes and 24 hours respectively. Then the absorbance was compared with each other and better antioxidant activity of the fruit juices was estimated. Results: The results obtained from the DPPH assay demonstrated that Grapes juice had better antioxidant activity than Pineapple and Guava juices. Conclusions: The present research demonstrates that although a number of popular beverages have evidence of antioxidant activity, there are clear differences in antioxidant potency. Some beverages with
lower potency would need to be consumed in much larger amounts to equal the antioxidant potency of
Grapes juice.
EnglishAnti-oxidant activity, DPPH (diphenyl picryl hydrazyl), Grapes, Guava, PineappleINTRODUCTION
Clinical trials and epidemiological studies have established an inverse correlation between the intake of fruits and vegetables and the occurrence of diseases such as inflammation, cardiovascular disease, cancer, and aging-related disorders 1 . The defensive effects of natural antioxidants in fruit and vegetables are related to three major groups; vitamin, phenolics and carotenoids and are believed to be the effective nutrients in the prevention of these oxidative stress related diseases 2,3. There is therefore a parallel increase in the use of methods for estimating the efficiency of such substances as antioxidants 4,5. One such method that is currently popular is based upon the use of the stable free radical 2,2-di(4- tertoctylphenyl)- 1-picrylhydrazyl (DPPH) which was an easy and accurate method with regard to measuring the antioxidant capacity of fruit and vegetable juices or extracts 4 .
Pineapple fruit is considered a highly nutritious fruit because it contains a high level of vitamin C, a natural antioxidant which may inhibit the development of major clinical conditions including heart disease and certain cancers 6 . The fruit also contains phenolic compounds and β- carotene 7,8, which constitute natural sources of antioxidants. Guava (Psidium guajava L.) fruit is considered a highly nutritious fruit because it contains a high level of ascorbic acid (50–300 mg/100 g fresh weight), which is three to six times higher than oranges. Phenolic compounds such as myricetin and apigenin 9 , ellagic acid, and anthocyanins 10 are also at high levels in guava fruit. The grapes has been well recognized worldwide for over 2,000 years as one among the edible sweet fruits and recognized for its wide spectrum of biological properties. Resveratrol (3,5,40- trans-trihydroxystilbene) is a natural phytoalexin abundantly found in grapes and red wine, which has potent antioxidant property 11. Thus, red fruit juices such as grapes and others like guava and pineapple have received attention due to their antioxidant activity. Whereas there are numerous phytochemicals consumed in our diet, polyphenols constitute the largest group and have attracted much attention due to their antioxidant properties 12. In fact, the potential health benefits of plant foods are commonly linked to their polyphenol content. Currently, there are a number of commercial ready-to-drink (RTD) polyphenol-rich beverages, which base their marketing strategies on antioxidant potency. However, to the best of our knowledge, data on the direct comparison of antioxidant activity of these widely available leading beverage products have not been obtained. It is of great interest to the general public to know the antioxidant capacity of the beverages that they consume. However, it should be cautioned that because of the inherent complexity of food matrices, the use of one antioxidant capacity method to determine antioxidant potency is ineffective. This is because antioxidants respond to different reactive species in different tests, which is partially attributed to multiple reaction mechanisms and reaction phases 13,14. The aim of the current study was to compare the antioxidant activity of three marketed fruit juices i.e., Grapes, Guava and Pineapple juices by DPPH assay.
Materials and Methods
Ready-to-Drink Polyphenol-Enriched Beverages
The following preparations are obtained from the local provisional market of Hyderabad: Grape, Guava and Pineapple of “Real Company”. All fruit juices were analyzed in late March or early April prior to their expiration dates as stated on their packages. All beverages were kept at storage conditions as specified on their labels prior to analyses. Analysis of Ascorbic acid content (AAC) The AAC was determined by the iodine titration method 15 or the RP-HPLC method: Waters C18 column (3.9×150 mm, 5µm particle size), mobile phase 5% acetic acid (Sd Fine Chem Limited, Mumbai), flow-rate 0.5 mL/min and 254 nm detection wavelength. Analysis of Total phenol content (TPC) TPC was determined using the Folin-Ciocalteu?s reagent 16 (LOBACHEMIE, Mumbai). Samples (0.3 mL, triplicate) were introduced into test tubes followed by 1.5 mL of Folin-Ciocalteu?s reagent (diluted 10 times with water) and 1.2 mL of sodium carbonate, 7.5%w/v (Virat Lab, Hyderabad). The tubes were vortexed, covered with parafilm and allowed to stand for 30 min.
Absorption at 765 nm was measured. If the sample absorbance exceeded 1, the sample was appropriately diluted to give reading less than 1. Total phenol contents were expressed in gallic acid equivalents (mg per 100 g fresh fruit).
Free Radical Scavenging Capacity:The free radical scavenging capacity was analyzed by the DPPH assay 17,18. 2,2-diphenyl-1- picrylhydrazy, DPPH (Santa cruz biotechnology, Inc) is a radical generating substance that is widely used to monitor the free radical scavenging abilities (the ability of a compound to donate an electron) of various antioxidants. The DPPH radical has a deep violet color due to its impaired electron, and radical scavenging can be followed spectrophotometrically by the loss of absorbance at 517 nm, as the pale yellow non radical form is produced 19.
The DPPH assay was typically run by the following procedure: In this method five dilutions of each fruit juice with two replicates were analyzed. Reaction solution was prepared by mixing 50 µL of diluted fruit juice with 300 µL of methanolic
DPPH solution (1mM) and the final volume was brought to 3 mL with methanol (Sd Fine Chem Limited, Mumbai). The solution was kept in dark at room temperature for 15 minutes. The absorbance (Ajuice) was read against the prepared blank (50 µL diluted fruit juice, 2950 µL methanol) at 517nm. A DPPH blank solution was prepared (300 µL of 1mM DPPH solution, 2.7 mL of methanol) and measured. Percent inhibition of DPPH radical was calculated for each dilution of juice according to formula: % Inhibition = [(ADPPH-Ajuice)/ADPPH×100] Where ADPPH is the absorbance value of the DPPH blank solution, Ajuice is the absorbance value of the sample solution. IC50, the concentration of antioxidant required for 50% scavenging of DPPH radical in the specified time period was derived from the % Inhibition vs Concentration plot. Results are shown in table and graphs.
Table 1: Table showing total phenolic content, ascorbic acid content and IC50 of fruit juices of the Real company
RESULTS
The results of total phenolic, ascorbic acid contents and IC50 of different fruit juices are summarized in Table 1. From the table the total phenolic content was found to be highest in Grapes juice 116.73 ± 0.14 mg/100g followed by Pineapple juice 95.20 ± 1.10 mg/100g and lowest in Guava 79 ± 0.06 mg/100g. The change in colorization from violet to yellow and subsequent fall in absorbance of the stable radical DPPH was measured at 517nm for various concentrations i.e. 50-250 μg/mL. The IC50 value for each fruit extract defined as the concentration of extract causing 50% inhibition of absorbance was calculated, since IC50 is a measure of inhibitory concentration, a lower IC50 value would reflect greater antioxidant activity of the sample. Antioxidant activity among the fruit samples was found to be maximum in Grapes having lowest IC50 value of 0.79 ± 0.34 mg/mL and minimum in Guava having highest IC50 1.71 ± 0.61 mg/mL as lower IC50 value would reflect greater antioxidant activity of the sample.
DISCUSSION
A polyphenol-rich food with health benefits has become a more common element in food marketing these days. The public is highly aware of the term “antioxidant”, which has been defined by the Institute of Medicine of the National Academy of Sciences as follows: “a substance in foods that significantly decreases the adverse effects of reactive species, such as reactive oxygen and nitrogen species, on normal physiologic function in humans.” Therefore, the marketing of many so-called “superfoods” is commonly based on their antioxidant potential. Multiple assays with different sensitivities and specificities for antioxidant activity are being used separately to justify health claims. Consumers have a difficult time distinguishing among the various antioxidant claims for widely available antioxidant beverages. Therefore, the present study was significant in comparing the most commonly available brands of beverages for antioxidant activity using the well-known and established laboratory methods for determining antioxidant capacity. Grapes juice had the highest antioxidant capacity and the most complete antioxidant coverage in vitro which may be contributed to its constituent, phytoalexin. The present research demonstrates that although a number of popular beverages have evidence of antioxidant activity in vitro, there are clear differences in antioxidant potency. The bleaching of the DPPH solution increases regularly with increasing amount of fruit in a given volume of solution. The bleaching action is mainly attributed to the presence of polyphenols and ascorbic acid extracted into the solution. The total phenolic content, IC50, and the ascorbic acid content of the fruit juices are summarized in Table 1, Graphs 1, 2 and 3. For a given amount of fruit, the higher the absorbance, the better is the reducing power. Correlation of IC50 with reducing power: DPPH assay measures the ability of the extract to donate hydrogen to the radical. In DPPH assay the lower the IC50 the better it is able to scavenge the radicals, particularly peroxy radicals which are the propagators of the autoxidation of lipid molecules and thereby break the free radical chain reaction 20. It is observed that grapes having low IC50, is a very potent radical scavenger. In terms of reducing power, grape rank highest but guava is significantly lower than that of pineapple. The high antioxidant potential (as characterized by low IC50 and high reducing power) of grape is attributed to its high TPC and AAC. The low antioxidant potential of guava (IC50 = 1.71 ± 0.61 mg/mL) is due its low TPC and AAC. Pineapple in spite of its relatively high TPC has low antioxidant potential (IC50 = 0.83 ± 0.24 mg/mL). Three possible reasons may be able to account for this: First, it has been reported that 21 reaction of DPPH with certain phenols such as eugenol and its derivatives is reversible, resulting in low readings for antioxidant activity (% disappearance). The second possible reason could be due to the slow rate of the reaction between DPPH and the substrate molecules 13 .The third possible explanation (for the relatively low reducing power) could be that certain phenols in the pineapple juice have a higher redox potential than that of other fruit juices. To clarify this anomaly further work is necessary. Finally, it is also observed that the antioxidant potential correlates well with AEAC.
CONCLUSION
For a body to maintain antioxidant level, external supplementation is necessary for healthy living. From the present research the order of the antioxidant activity in the fruit juices was found to be Grapes>Pineapple>Guava. These fruits can be used as alternative source of natural antioxidant rather than synthetic antioxidant like BHT (Butylated hydroxytoluene) and BHA (Butylated hydroxyanisole) because of carcinogenicity.
ACKNOWLEDGEMENT
We thank everyone who supported this study
Englishhttp://ijcrr.com/abstract.php?article_id=2304http://ijcrr.com/article_html.php?did=23041. Swartzberg J, Margen S. Eat, Drink, and be Healthy-The HarVard Medical School Guide to Healthy Eating. Am J Epidemiol 2001;154(12):1160.
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4. Sanchez-Moreno C. Review: Methods used to evaluate the free radical scavenging activity in foods and biological systems. Food Sci Tech Int 2002;8(3):121-137.
5. Schwarz K, Bertelsen G, Nissen LR, Gardner PT, Heinonen MI, Hopia A, et al. Investigation of plant extracts for the protection of processed foods against lipid oxidation. Comparison of antioxidant assays based on radical scavenging, lipid oxidation and analysis of the principal antioxidant compounds. Eur Food Res Technol 2001;212:319-328.
6. Diplock AT. Antioxidants and disease prevention. Molecular Aspects of Medicine 1994;15:293-376.
7. Gardner PT, White TAC, McPhail DB, Duthie GG. The relative contributions of vitamin C, carotenoids and phenolics to the antioxidant potential of fruit juices. Food Chemistry 2000;68:471-474.
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12. Proteggente AR, Pannala AS, Paganga G, Van Buren L, Wagner E, Wiseman S, et al. The antioxidant activity of regularly consumed fruit and vegetables reflects their phenolic and vitamin C composition. Free Radical Res 2002;36:217-233.
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19. Seeram NP, Aviram M, Zhang Y, Henning SM, Feng L, Dreher M, et al. Comparison of Antioxidant Potency of Commonly Consumed Polyphenol-Rich Beverages in the United States. J Agric Food Chem 2008;56:1415-1422.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesCAREER, JOB SATISFACTION AND ITS EVALUATION METHODS
English7479Muhammad FarzanjouEnglishJob is among the issue that has often been engaged by humans mind, governments and nations. Although job and occupation is apparently associated with economic-subsistence aspect of humans, it is closely linked to their personal, familial, social, political and cultural dimensions. Job satisfaction is a filed within which social psychological, sociological, economic and political and educational sciences perspectives have been mentioned. Today, there are thousands of jobs and professions in each country which people are involved in and thereby continue their life. What always attracts the attention of psychologists and social sciences thinkers are people's job satisfaction and the effects of this satisfaction in their spirits and working productivity. If one is not interested in his/her job, one's creativity and talent will not be flourished in one's work field and then will be affected by fatigue, depression and frustration and his/her work will be inconclusive, and hereby the community will be affected. This article aims to consider this issue.
Englishjob, job satisfaction, factors of job satisfaction, methods of assessment.1. INTRODUCTION
1.1. Definition of Job
Job literally means gets someone to work and what is the cause of engagement. The individuals will actively participate in the production processes and services through employment and then receive cash or material rewards (1). Job and work is a physical or intellectual activity which is directed on production and services. Generally, job is an activity which is asked for and hence is paid by (2). In summary, it can be said that job is the work a person is involved with and through which perform his/her duties and gaain his livelihood. In another definition, job is a group of similar positions in an institution, office or workshop in which the quailed people are able to take this opportunity and carry out their duties (3).
1.2. Definition of Job Satisfaction
Job satisfaction is a set of feelings and beliefs that people have about their current jobs (4). Job satisfaction is one of the important factors in job success, a factor that increases efficiency and also feeling of personal satisfaction (5). Job satisfaction means to be interested in the situations and requirements of a job, a situation in which a work is done and the reward for which is received (6). According to the above-mentioned contents, it can be indicated that job satisfaction is feelings of fulfillment and satisfaction one have of him/herself and the pleasure is therefore achieved and consequently will be encouraged and attached to his/her job. Job satisfaction is a desirable, emotional and positive state that is resulted from job assessment or job experiences. It is a concept that has various dimensions, aspects and factors which must be taken into consideration as a whole. Of these factors are the characteristics of employer and employee, type of work, work environment and human relations in working (7). V.E.Fisher and J.V.Hanna maintain that jib satisfaction is an internal factor and believes that it is a type of emotional compatibility with job and job conditions, that is if the regarded job provides the satisfaction and desirable pleasure, he/she will be happy with his/her job, and conversely, if the regarded job does not provide the satisfaction and desirable pleasure, he/she will be disappointed and is going to change it (8). R. Hoppock maintains that job satisfaction is a complex and multi-dimensional concept and is associated with mental, physical and social factors. Only one factor is not led to job satisfaction, but a specific combination of a set of various factors cause an employer is satisfied with his/her job in a specific moment of time, and tell him/herself that he/she is satisfied with his/her job and enjoys it (9). it is found out from the above0definitions about job satisfaction that this concepts accounts for the positive and feelings and attitudes which one has towards his/her job. When it is said that someone has high levels of job satisfaction (10), it means he/she generally likes his/her job and considers a great value for it and takes it into account as a positive issue, in short, he has a good and desirable feeling towards it (11).
2. Factors of Choosing Job
Job is an issue that is not randomly chosen, but it requires many factors, including:
2.1. Physical Condition
Each job requires a certain physical characteristics. A great size and strength is required in some jobs, while these features may prevent doing some tasks in some jobs. Also, having hand and foot health is necessary in some jobs, while in others, lacking or defecting limbs and other organs does not create any problem.
2.2. Talent
One of the important factors in job selection and continuing a successful employment is talent. Talent means being equipped with, readiness and ability to perform a certain job, that is one's innate ability that helps him learn and accelerates it. Hence, talent predict the method and amounts of learning in various fields in the future, and the one who is gifted in a certain area will be more benefitted with his/her experiences in that regard.
2.3. Interest
Interest means having desire, willingness and desire to acquire something. Also, the pleasant feeling, desire or curiosity towards something or concept is called interest. Interest is an important impetus for effort and human's activity. Success in any job requires having interest.
2.4. Personal and Social Facilities
In addition to the above-mentioned materials, other factors such as personality, realism, environmental facilities and community needs has an important effect in choosing job (12). In short, it can be said that personal factors (such as physical status, talent, interest and personality traits), social factors (such as family pressure, social and cultural values, the amount of facilities in a community and the opportunities that are available to individuals), economic factors (such as poverty and unemployment) as well as inheritance and gender are effective in job selection.
3. Factors of Job Satisfaction
Researchers have long been in search of fundamental causes of job selection in offices and organizations. They have been able to achieve a string of constant factors associated with job satisfaction; however, not a comprehensive empirical model has yet been achieved. Several factors can be briefly pointed out that are more important in this field. W. Porter and M. Steers pointed out to four factors as follows:
Overall factors of organization: i.e. variables that are widely applicable to most employees, such as salary and promotion opportunities.
The immediate causes of occupational environment: variables representing occupational groups, such as supervision method and quality of relationship with colleagues, working conditions and workplace.
Content factors or actual occupational activities, such as occupation territory (of diversity, autonomy and responsibility) and role clarity.
Individual factors: the characteristics that make a person distinct from another as well as age, duration of service and character (self-confidence, determination and maturity) (13).
E.A.Locke summarizes the most important factors influencing job satisfaction as follows:
Mental precarious work that a person can be successfully compatible with (success in coping with the work).
Personal interest to the job itself in that the more one is interested in a job, the more would be his/her satisfaction.
A work that is not physically too boring, that is the more is a person tired, the less will be his/her satisfaction and the less one is tired, the more will be his/her satisfaction.
Reward for performance would be fair, informative and consistent with one's demand.
Working conditions that would be consistent with physical needs and contribute to career goals.
Feeling of self-esteem from the employed, the more one feels respect from others, the more will be his/her satisfaction.
Factors in workplace that facilitate the need for occupational values, such as increased pay and promotion (14).
4. Results of having job satisfaction Being aware of significant results of job satisfaction is as important as being aware of what leads to satisfaction. These results are as follows:
4.1. Satisfaction and Service Renunciation
Job satisfaction and service renunciation is related with each other. V.H.Vroom found out that the correlation range between these two variables in various studies is from 25% to 42%.
Revisionists in recent decades, which studied the relationship between job satisfaction and job renunciation, reported that there is a negative relationship between them, that is; if employees are satisfied with their job, they will not leave their job and vice versa, that is if they are not satisfied with their job, they will leave their job. A relatively similar report was presented by Lock in 1976.
4.2. Job Satisfaction and absence in working
The evidence shows that there is a balanced and an inverse relationship between job satisfaction and absence of employees from their workplace. Vroom showed in various studies that the compatibility range is from 14% to 38%. This study was confirmed by Porter and Stirz , etc.
4.3. Satisfaction and Performance
One of the most controversial issues in the field of job satisfaction is its relationship with performance. Three hypotheses have been proposed in this regard. 1. Satisfaction is led to performance. 2. Performance is led to satisfaction. 3. Reward acts as an intermediate between performance and satisfaction. The first two hypotheses are weakly supported, but the third one, based on which reward acts as an intermediate between performance and satisfaction, has been more strongly supported. Prior performance is resulted in receiving internal reward (feeling of personal prosperity) and external reward (wage and promotion). This reward, by itself, enhances one's performance in the future and also is effective in increasing one's job satisfaction. Vroom found out in his studies that there is a positive relationship between job satisfaction and amount of efficiency and performance. Steers and Porter mentioned in their books that the more an employee and worker's occupational motivation and the more their attitude towards their jobs (that is, to be more satisfied with his/her job), the more will be their performance and vice versa, that is, the less is motivation and positive attitude towards one's job (the less job satisfaction), the less will be one's performance.
4.4. The Effect of Satisfaction on Organization
The analyses indicate that when employees of an organization are satisfied with their jobs, their organization are faced with positive effects and acts as an effective and useful organization. In addition to above-mentioned issues, job satisfaction has some other results. The employees who are entirely satisfied are less intended to submit complaint and are more acquired with physical and mental health and have a more longitude; they learn new duties related to their job more swiftly and are acquired with less occupational accidents (15).
5. SUGGESTIONS
According to the above contents, the following works are suggested for increasing job satisfaction in an organization: ? Making employees interested in their job to flourish creativity and innovation. ? Preventing from depression and frustration in employees. ? Creating factors that increase efficiency and satisfaction in organization. ? Creating significant motivation and interest for the effort and willingness of people to successfully perform the job ? Being respected from other people to provide greater satisfaction.
6. CONCLUSION
By reflecting and hesitating on what was mentioned and by considering the strategic issues and the coherent guidance of organization, which reminiscent of undeniable and important role of managers which is certainly highly effective in victory or defeat of programs, creating " job satisfaction" in a person depends on various factors which together are achieved to optimal result, and somehow lack of one factor in a person is led to one's job dissatisfaction; factors such as income level, nature of work and its social status, organizational prestige and authenticity of job promotion, job security, lack of role ambiguity, physical working condition, structure and organizational culture and relationship with colleagues, paying attention to one's personality traits, performance evaluation, fitness, flexibility and innovation. Finally, it can be said that job satisfaction is totally a positive and pleasant feeling that one has about his/her job. More generally, scientists maintain that social factors, working environment and the nature of working are effective in job satisfaction. All theories of job satisfaction somehow emphasize on meeting people's needs, whether physical or mental and takes the demands and expectations of employees as an important thing.
Englishhttp://ijcrr.com/abstract.php?article_id=2305http://ijcrr.com/article_html.php?did=23051. Abdollah Shafiabadi, "professional and career counseling and guidance", Roshd Publications, P. 3, Tehran, Iran, 1997.
2. Boures. A. Shertzer, "occupational life planning and review", translated by Zandipour, P. 207, Tehran, Iran, 1990.
3. Abdollah Shafiabadi," Professional ad career guidance and counseling", Roshd Publications, P. 40, Tehran, Iran, 1997.
4. George. M. Jennifer & Jones. R. Gareth,”Organizational Behavior Understanding and managing” ,Addison Wesley, P. 74, New York,U.S.A,1999.
5. Abdollah Shafiabadi," Professional ad career guidance and counseling", Roshd Publications, P. 123, Tehran, Iran, 1997.
6. Boures. A. Shertzer, "occupational life planning and review", translated by Zandipour, P. 207, Tehran, Iran, 1990.
7. Hellriegel. Don , Woodman. W. Richard,” Organizational Behavior” ,South - Western College Publishing An International Thomson Publishing Company,P. 53-55, 1996.
8. Gholam Abbas Tavasoli, "working and occupation sociology", SAMT Publications, PP. 166-170, Tehran, Iran, 1996.
9. Khadijeh Safiri, "sociology of women employment", Tabyan Publications, P. 76, Tehran, Iran, 1998.
10. Abbas Mahmoudzade, Armen Mehroujan, "organizational behavior of contingency perspective", Allameh Tabatabaie University Press, P. 274, Tehran, Iran, 1996.
11. Abdollah Shafiabadi, "career and academic guidance", publication center and Payam Nour University Press, Tehran, Iran, 1991.
12. Abbas Mahmoudzade, Armen Mehroujan, "organizational behavior of contingency perspective", Allameh Tabatabaie University Press, P. 374, Tehran, Iran, 1996.
13. Iraj Soltani, "Productivity of Human Resources", Arkan Publications, Isfahan, Iran, 2006.
14. Leap. L. Terry, Crion. D. Michael,” Personal Human Resource Management”, Macmillan Publishing, P. 366-367, New York, 1989.
15. Norollah Khalilzade, " study of the factors effective in job satisfaction and dissatisfaction of students and teachers of Payame Nour University of Oroumieh", Faculty of Psychology and Educational Sciences, Tehran University, PP. 21-25, Tehran, Iran, 1997.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareDRUG- RESISTANCE PROFILE OF NEW PTB PATIENTS WITH OR WITHOUT HIV INFECTION
English8084Niladri Sekhar DasEnglishTuberculosis is the commonest opportunistic infection in persons infected with human immunodeficiency virus (HIV). HIV is the most powerful risk factor for reactivation of latent tuberculosis infection to active disease. Emergence of drug resistant isolates of M. tuberculosis (Mtb) highlights the need for continuous monitoring of drug resistance to anti tuberculosis drugs. The aim of our study was to see the anti tubercular drug resistance profile of M.tb in HIV seropositive and seronegative patients. Material and methods- We enrolled 100 new pulmonary tuberculosis (PTB) patients. This study was carried out at Department of Microbiology MGIMS, Sevagram, Wardha Maharashtra from 2007-2009. For every patient staining and culture was done from sputum sample by conventional method. After the identification of isolates as M.tb drug sensitivity testing was done. HIV antibody status was identified in every patient by using rapid and ELISA test. Results were analyzed using SPSS software. Results: There were 8 patients who were HIV seropositive out of total 35 culture positive cases. No resistant patterns were detetected in case of HIV +ve patients with TB infection. Out of 27 HIV seronegative cases with TB infection, 4 cases showed mono-resistant to streptomycin. No multi drug resistant (MDR) strain was detected neither in HIV positive nor in HIV negative cases. Conclusion: HIV infection may not be associated with drug resistant tuberculosis. However due to high prevalence of HIV–TB infection in the country, monitoring of drug resistance in M. tuberculoisis isolates needs prioritization to ensure success in national tuberculosis control programme.
EnglishHuman immunodeficiency virus (HIV), Pulmonary tuberculosis (PTB), Mycobacterium tuberculosis (Mtb), Multi drug resistant (MDR)INTRODUCTION
Tuberculosis (TB) is one of the most serious diseases that physicians have recognized and grappled with through the millennia due to its high risk of person to person transmission, morbidity, and mortality. Currently there are 9.2 million new cases of tuberculosis every year with 1.7 million deaths occurring worldwide1 . Despite the discovery of the tubercle bacillus more than a hundred years ago, and all the advances in our knowledge of the disease made since then, tuberculosis still remains one of the major health problems facing mankind, particularly in developing countries. In India each year, 1.9 million new cases of TB occur in the country, of which about 0.8 million are infectious new smear positive pulmonary TB cases2 . The situation is expected to worsen due to the emergence of multi-drug resistant tuberculosis (MDRTB).
Development of drug resistance in M. tuberculosis isolates may adversely impact the management of the disease. Inadequate therapies and indiscriminate use of antibiotics causes such emergence of multi drug resistant bacilli and HIV/AIDS pandemic resulting in accumulation of pool of individuals who are more susceptible to TB have worsened the TB scenario. A High HIV seroprevalence among newly diagnosed TB patients has been reported in India.3, 4 . On the global situation of drug resistance in M.tb , first comprehensive study was conducted by the Research and Surveillance unit, Global TB Programme, WHO5 and was observed that, the rates of acquired resistance were invariably higher than primary resistance for each drug and multi drug resistance was more common when resistance was acquired rather than when it was primary. The increasing prevalence of multidrug resistance in several parts of the world including India has been one of the major reasons for declaring tuberculosis control as a global emergency by WHO6,7 . Several outbreaks of MDR-TB showed the importance of continuous monitoring of drug resistance for treatment of TB patients and initiating adequate public health measures. The present study was undertaken with the objective of comparing the anti TB drug resistance profile of MTB isolates in new PTB patients with or without HIV infection.
MATERIAL AND METHODS:
Among 100 suspected tuberculosis patients study was done at Microbiology Laboratory in MGIMS, Sevagram between 2007 to 2009 with due permission from the institutional ethical committee. Sputum was collected from each individual, smear was made and staining was done by Ziehl–Neelson method. For isolation rest of the sample was decontaminated by NALC-NaOH method8 and inoculation was done on 2 LJ slopes. LJ bottles were observed weekly till the colonies grew sufficiently for identification whereas negative LJ culture was noted if there was no growth after incubation at 370 c for 8 wks. Isolates were identified later by conventional biochemical methods9 After the identification of the strain as MTB, drug sensitivity testing was performed against 1st line drugs by LJ proportion method described by Canetti et al 1969 10. For Drug sensitivity testing stock and working concentration of drugs were made according to Guidelines of TRC manual 200611. The concentration of drugs used were Streptomycin 4 µg/ml, INH 0.2 µg/ml, Rifampicin 40 µg/ml and Ethambutol 2 µg/ml. Before drug sensitivity testing by LJ proportion method, drug containing LJ slopes was made. At first preparation of standard suspension 11 and dilution was made. 0.1 ml of each of the two bacterial dilutions, 10–3 mg/ml and 10–5 mg/ml were inoculated on two slopes of LJ medium without drug and one slope of medium with drug. The standard strain of M.tuberculosis, H37Rv was tested with each batch of sensitivity testing. All the LJ slopes for DST were placed in the incubator at 37 °C. Reading and reporting of the LJ slopes were done according to the reference10 Detection of HIV status in every patient was done by rapid method (TRIDOT) using serum which was collected in a plain bulb. If any sample was found to be positive by rapid method, confirmation was done by ELISA (Vironostika) confirmed by second ELISA at NARI Pune.
RESULTS:
We got 35 total MTB isolates no NTM (Non tubercular mycobacterium) was isolated. Among 35 MTB isolates 87.9 percent (29/33) from smear positive and 9 percent (6/67) from smear negative patients were obtained. Of these 8 isolates were HIV seropositive and 27 HIV seronegative (Table no 1). Among the 35 total MTB isolates, monoresistant was found with Streptomycin that was 11.4% only in HIV seronegative patients (Table no 1). Resistance to any 1st line drug was seen (7/35), in 20% isolates. Drug resistance to at least one drug was found in (7/27), 25.9% isolates in HIV seronegative cases whereas no resistant strain was isolated even for a single drug in HIV+ve cases however in HIV seropositive cases we got (8/8), 100% isolates sensitive to all four drugs. The prevalence of polyresistance (resistance to two or more drugs, but not both isoniazid and refampicin) was found to be (3/27), 11.11% only in HIV seronegative with TB infective patients. Neither individual drug resistance nor the MDR or polyresistance was found to differ significantly in the HIV infected and uninfected TB patients.
DISCUSSION
Emergence and spread of drug resistance in M.tuberculosis is a serious threat to tuberculosis control programme because patients with drug resistance bacilli respond less readily to therapy than those with sensitive bacilli, resulting in preferential spread of the drug resistant bacilli in the community. Drug resistance develops either due to infection with a resistant strain, or as a result of inadequate treatment such as when a patient is exposed to a single drug, or because of selective drug intake, poor compliance, use of inappropriate non-standardized treatment regimens, irregular drug supply, poor drug quality, or rarely erratic absorption of medications 12 . Though the development of drug resistance in India was noted since the beginning of the chemotherapeutic era, it was based on clinical perception and several isolated reports, which failed to give an idea of the national situation as a whole. In this direction the first definite step was taken in 1965-67, when the ICMR conducted two surveys to estimate the prevalence of drug resistance13 . In this study resistance to Isoniazid ranged from 11-20 per cent, to Streptomycin 8- 20 per cent and to both drugs 4-11 per cent. The second study showed resistance to Isoniazid from 15-69 per cent, to Streptomycin 12-63 per cent and to both drugs from 5-58 per cent. However these studies were carried out in the pre-Rifampicin era. Subsequently after about two decades (1980?s), 11 other studies from different parts of the country reported similar resistance to Isoniazid and Streptomycin like those in the earlier studies with the notable exception of Rifampicin resistance being observed in 10 of the 11 publications and the level of MDRTB in all the centers was observed but it was less than 5 per cent. According to third Global Report total prevalence of drug resistance among new cases in India (Wardha) was 19.8% and MDR was 0.5 %. Several small surveys conducted across the country have shown the prevalence rates of MDR-TB in the country at around 3% among new cases, and 12% among retreatment cases. A large scale population based survey in the states of Gujarat and Maharashtra has also indicated similar resistance levels (new- 3% and retreatment- 12- 17%) 13. We found that prevalence of resistance to isoniazid and streptomycin was 0 and 11.4 percent and that to both drugs were 2.8%. The issue of whether infection with HIV is a risk factor for drug resistant tuberculosis still remains unanswered since results of some studies supported this hypothesis while some did not. A study conducted in newyork city 14 revealed that HIV infected TB patients were significantly more likely to develop resistance to at least one drug and MDR than those without HIV infection. No significant difference was found in drug resistance between HIV seropositive and HIV seronegative tuberculosis patients in our study, which is in accordance with the findings from other studies in Europe 15, 16 . In conclusion, we didn?t find any kind of drug resistance in anti-tubercular drug especially in case of HIV seropositive cases. In view of the conflicting results from other studies and high prevalence of HIV-TB in our country, we feel that time to time monitoring of anti-tuberculosis drug resistance pattern in TB patients in general and HIV seropositive tuberculosis patients in particular would provide important data, which may be crucial for the control of tuberculosis.
Englishhttp://ijcrr.com/abstract.php?article_id=2306http://ijcrr.com/article_html.php?did=23061. Global tuberculosis control: surveillance, planning, financing: WHO report 2008. Geneva: World Health Organization (WHO/HTM/TB/2008.393).
2. TB India 2008. RNTCP Status Report.
3. Paranjape R S ,Tripathy SP, Menon PA, Mehendale SM, khatavkar P, Joshi DR.et al. Increasing trend of HIV seroprevalence among pulmonary tuberculosis patients in Pune, India .indian J Med res 1997;106: 207- 11
4. Tripathy S, Joshi DR, Mehendale SM , Menon P, Joshi AN, Ghorpade SV, et al.Sentinel surveillance for HIV infection tuberculosis patients in India. Indian J tuberc 2002; 49: 17-20.
5. Cohn DL, Bustreo F, Raviglione MC. (1997). Drug resistant tuberculosis: review of the worldwide situation and the WHO/IUATLD global surveillance project. Clin Infect Dis 1997; 24(1): 121-130.
6. Anti tuberculosis drug resistance in the world. The WHO/ WATLD global project on anti tuberculosis surveillance .Geneva,Switzerland;1997.WHO/TB/97.229.
7. Anti tuberculosis drug resistance in the world ,report No.2.WHO/CDS/TB/2000.278
8. Procedure for isolation and identification of mycobacterium CDC manual1975.
9. Laboratory services in tuberculosis control .Part 3-Culture /WHO/98.258,1998.
10. Canetti, G., Fox, W., Khomenko, A., Mahler, H.T., Menon, N.K., Mitchison, D.A., et al. Advances in techniques of testing mycobacterial drug sensitivity, and the use of sensitivity tests in tuberculosis control programmes. Bulletin of the World Health Organisation, 1969. 41, 21-43.
11. Bacteriological methods and Laboratory diagnosis of Tuberculosis TRC Chennai 2006.
12. Behera D. Drug Resistant Tuberculosis In India – Is it a matter of concern? Ind J Tuberc 2007; 54:105-109.
13. Multi-drug resistant and Extensively drug resistant TB in India, Consensus statement on the problem, prevention, management and control, From the consultative meeting of national experts organized by the TB Research Centre, ICMR, Govt. of India, on 14-15 September 2007.
14. Gordin FM, Nelson ET,Matts JP, Cohn DL, Ernst J,Benator D,et al. The impact of human immunodeficiency virus infection on drug resistant tuberculosis . Am J.Respir Crit care Med 1996; 154: 1478-83.
15. Migliori GB, Centis R, Fattorini L, Besozzi G, Saltini C, Scarparo C, et al. Mycobacterium tuberculosis complex drug resistance in Italy. Emerg Infect Dis 2004;10:752-3
16. Shafer RW, Chirgwin KD, Glat AE, Dahdouh MA, Landesman SH, Suster B. HIV prevalence , immunosuppression , and drug resistance in patients with tuberculosis in an area endemic for AIDS.AIDS 1991;5:399-405.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesBACTERIOLOGICAL APPLICATIONS OF QUANTUM DOTS
English8594Nithish.U.SEnglish Sarah SunithaEnglishQuantum Dots are nanocrystals which are fluorescent in nature and their unique optical properties depend on their size. Quantum Dots have replaced conventional fluorophores which have disadvantages like photobleaching, narrow absorption spectra, stability and short period of fluorescence. Due to the possibility of conjugating the Quantum Dots to various types of bio molecules like streptavidin, antibodies, mannose etc there have been numerous applications in the detection, enumeration and differentiation of various bacteria. Quantum Dots can be applied to various matrices like food, water, tissue and blood samples. Quantum Dots have been used to detect pathogenic bacteria like E.coli, Staphylococcus aureus, Salmonella typhimurium, Bacillus anthracis, Listeria monocytogenes and Mycobacterium tuberculosis. Quantum dots can detect bacteria at the levels of 103- 10 CFU/ml in contaminated water samples. The minimum time for detection of bacteria using Quantum Dots ranges from 15 minutes to 2 hours. The major hindrance in using the Quantum Dots is its cost of production. This review summarises the properties, synthesis and applications of Quantum dots in the detection of bacteria which can have major implications in food and water safety evaluation, diagnosis of bacterial diseases and environmental enumeration of bacteria.
EnglishQuantum Dots, Optical properties, Detection, EnumerationINTRODUCTION
Quantum Dots (QD) are highly crystalline semiconductor nanocrystals. These are crystals whose dimension is less than that of the Exiton Bohr radius [1].Due to their nanoscale size; they have discrete electronic energy state giving rise to unique optical and electronic properties. They also possess the property of size dependent photo-emission which is due to the phenomenon of Quantum Confinement [2]. Their stability against photobleaching has attracted many researchers to develop bio-imaging and Quantum computing techniques. Due to their unique properties they have wide range of applications in electronics, computing and biology. QDs are made up of elements like Cadmium, Selenium, Indium, Tellurium, Phosphorous, Sulphur etc. These elements make up the core of a Quantum dot, which are then coated with a shell made of material of higher band energy. The outer shell also forms a protective layer on the Quantum dots. To make QDs biologically compatible one or more layer of organic polymers are added and are conjugated with organic ligands. The diameter of QDs varies from 1-20nm which consists of 100-100,000 atoms [2]. Due to their distinctive properties, QDs have improved the sensitivity of cellular analysis and molecular detection by few folds. The Quantum Dots were first studied by A.I. Ekimov and A.A. Onushchenko [3]. The size of microscopic CuCl crystals grown in transparent dielectric matrix varied from tens to hundreds of angstroms. Later high quality CdSe, CdS and CdTe semiconductor nanocrystals were synthesized based on pyrolysis of organometallic reagents [4]. The particles sizes were as small as 20 angstroms in diameter. Syntheisis of water soluble Quantum dots paved way for their conjugation to bio molecules and thereby opened up different applications in biology [5, 6]. In Quantum Dots, the energy levels of the valence electrons are discrete and have large differences. Thus they show the characteristics of an atom and hence they are also called artificial atoms [8]. The reduction in the size of the crystal brings the valence electrons closer thereby increasing their overall energy. This leads to splitting up of different electron levels giving it a discrete electron energy level system and also increasing their band gaps. This results in merging of valence band and conduction band making it a very good conductor of electricity. The splitting of energy level leads to excitement of the electron by absorption of energy and transition to higher energy levels. But these electrons return to their ground state by emitting a photon of energy either equal or lesser than that of the absorbed energy. The emitted photon has lesser energy when the electron rests in a metastable level for a brief period of time. This gives QDs their fluorescent property making them applicable in imaging and detection of molecules or cells. Thus as the size reduces, the gap between energy levels broadens up and the electron has to return from a much higher level which increases the emitted photon energy. Therefore as the size reduces, the emitted photon shifts from the red region of visible light spectra to the blue region. This gives rise to the unique fluorescent emission of the Quantum Dots.
PROPERTIES
The exceptional properties of the Quantum Dots like photo stability, broad range of absorption spectra, ease in change of emission spectra, sensitivity, brightness and ability of coupling with biological molecules, make them very useful in the field of biological imaging of different types of cells. Quantum Dots emitting different colours can be excited by monochromatic light. The time taken to fluoresce upon excitation is very less compared to conventional fluorophores and this property lasts longer for a given Quantum Dot. The Quantum Dots are very stable and resistant to photobleaching and overcome the disadvantages exhibited by fluorophores, which are important for imaging applications. SYNTHESIS QDs can be synthesized by colloidal synthesis, Efield method or by fabrication through etching or self-assembly.
Colloidal chemistry method
This method is also called Hot-Injection method [2].Here Quantum Dots are prepared as a batch by quickly adding appropriate amount of precursors into hot solvent and organic ligand system. First the precursors are decomposed to monomers by chemical or physical means like temperature. This results in nucleation process where the monomers starts binding to each other. This reaction is fast in the beginning and slows down in the end due to change in concentration.
E-field technique
In this method, Quantum Dots are synthesised between the interfaces of heterojunction of monomers [8]. On applying an electric field on the interface, 2D Electron Gas [2DEG] is created. Later these charges are confined to the interface by external devices. Thus the voltage and the 2DEG permit rearrangement of atoms, finally giving a Quantum Dot at the interface.
Fabrication
Etching or Lithographic method
In this method, first a Quantum Well or Double Barrier Heterostructure [DBH] are grown by the Epitaxial Method. Then pillars are etched out of it and they are further etched to give nanocrystals like Quantum Dots [8]. These pillars have the dimension of a Quantum Wire, which is then metalized on each terminal. Application of electric current on the terminals of Quantum Wire leads to the breakdown of the Quantum Wire and formation of Quantum Dots.
Self-assembly growth technique
Here Quantum Dots are formed as an out growth in the Epitaxial technique [8]. The effect is similar to the condensation of water droplets on cold surface. Generally, the epitaxial growth proceeds by forming a layer of atoms at once. But lattice mismatch and difference in surface energy between the atoms induces the formation of islands, thus Quantum Dots are formed. It is seen that bare Quantum Dots are unsuitable for biological application. This is because; they are insoluble in water and toxic due to their heavy metal composition, small size and active surface [9]. To overcome these problems, the surface of Quantum Dots is treated with stabilizing and coating substances. Most of the coating substances are organic polymers [10] which are amphiphilic in nature, also making them water soluble and easy to apply for biological analysis by conjugating with proteins, antibodies and other molecules. Additionally, it is observed that coating provides better photo stability [11]. It is also seen that some bio molecules like proteins can be effective nucleating and stabilizing agents for Quantum Dots [12].
APPLICATIONS OF QUANTUM DOTS IN BACTERIOLOGY
The superior optical properties of Quantum Dots entail them for numerous applications in labelling of cells. They are highly suitable for labelling, as the intense emission of light helps in detection of even a small number of the cells. They find applications in enumeration of bacterial cells in various matrices, detection of pathogens, food contaminants, and in the study of micro flora in bio films.
Enumeration
Here the number of bacterial cells can be enumerated by relating the intensity of light emitted by the Quantum Dots to the cell count. For example, Alteromonas species inhabiting the surface of small marine animals like Copepods were enumerated using QDs [13]. The CdSe Quantum Dots were conjugated with streptavidin so that QD can specifically bind to the anti-antibody against Alteromonas species. It is also possible to detect wide range of bacteria from a sample such as sewage water using water soluble Quantum Dots [14]. Quantum Dots can also be used to detect a particular strain of a bacterial species as supported from the result of an experiment where mannose conjugated CdS Quantum Dots could detect a strain Escherichia coli producing FimHlectin which binds specifically to mannose [15]. In a study on fluorescence detection of count of Escherichia coli and Staphylococcus aureus using Quantum Dots, spectrofluorometer was used for roughly counting the cells [16]. The low detection limit for this method was in order of 102CFU/ml. In a different study the same detection limit was obtained when the Quantum Dots were labelled to bacteria covalently using glutaraldehyde [17]. It was also seen that there was a linear relationship between fluorescence intensity and total bacterial count. By using Quantum Dots it is possible to distinguish between wild type and auxotrophic strain of a bacteria belonging to same species. This technique involves conjugating Quantum Dots with the biomolecule needed for bacterial growth but which cannot be synthesized by the bacteria. This method is applied for detection of purine auxotrophs of Bacillus subtilis and Escherichia coli where Quantum Dots conjugated with AMP were taken up preferentially by purine-auxotrophs rather than the wild type [18]. The Quantum Dots can also be coupled to the antibodies used in ELISA which can give us an intense signal even in a minute concentration of antigen. This method was applied for detection of Escherichia coli in water samples where the target bacteria were separated by antibody coated magnetic beads and then detected by ELISA using Quantum Dots conjugated to secondary antibodies [19]. Thus Quantum Dot based enumeration of bacteria can be rapid and sensitive compared to the conventional methods.
Detection of pathogens
Quantum Dots have better optical properties than other dyes or fluorophores; it is widely applicable in detecting pathogens from various clinical samples and to detect a particular pathogen from a cluster of bacteria. There are number of studies where Escherichia coli and Salmonella typhimurium were detected simultaneously using antibodies against these two species and conjugating different antibodies with Quantum Dots emitting light of different wavelength [20]. Multiplexed detection was also applied for Escherichia coli and Salmonella enteritidis [21]. Denatured Bovine serum albumin stabilized Quantum Dots were conjugated to secondary antibodies for detection of Escherichia coli and Listeria monocytogenes [22]. This type of conjugation to the secondary antibodies gives us an advantage of universal usage of the antibody for the detection, ease in their production and circumventing the tedious process of labelling individual primary antibodies. The interactions between antibody and antigen are weak and these are further weakened by conjugation of bulk substance like Quantum Dot. This reduces the staining property of Quantum Dots conjugated to antibodies, hence, it is preferred to conjugate small molecules like biotin to antibody and then bind them to Quantum Dots conjugated to streptavidin. The above method was also adopted for detection of E.coli [23] and Listeria monocytogenes where the limit of detection was found to be 2-3 CFU/ml with a detection time of 1.5 hrs. [24]. In another study Mycobacterium bovis BCG and Mycobacterium tuberculosis was detected using genus specific antibody and biotinylated antiantibody which would bind to streptavidin coated Quantum Dots [25]. The result gave a limit of detection of 103 bacteria/ml. This method was also applied for detection of Escherichia coliO157:H7 strain with a limit of detection of 10 3 CFU/ml with a detection time less than 2hrs [26]. In one of the studies Escherichia coli was imaged using organic acid stabilized Quantum Dots [27]. Here the Quantum Dots were internalized by the bacteria giving rise to fluorescence signal. It was found that none of the other bacteria interfered with the test. Quantum Dot labelling of the cells also aids Flow cytometric separation and analysis of pathogenic bacteria from other non-pathogenic ones. This method is a fast and effective technique for separation. In a study pathogenic Escherichia coli strain O157:H7 were separated from non-pathogenic bacteria using Quantum Dots and Flow cytometer [28]. The limit of detection was 1% of the pathogenic strain in the mixture of cells.
It is also possible for detection of strain and metabolism specific bacteria using Quantum Dots conjugated to a biomolecule which might bind to the receptor on the particular strain or may be used as a substrate for a metabolic pathway [29]. In another study different antibodies are conjugated to Quantum Dots of different emission peaks for multiplexed analysis of Bacillus anthracis spores and Yersinia pestis by Flow cytometry [30]. It also possible to detect one spores of Bacillus anthracis from other similar bacterial spores by conjugating Quantum Dots by BABA peptides [31] or a short peptide chain from gamma-phage lysine protein [32]. This analysis can be performed using methods like Flowcytometry, Confocal laser scanning microscopy and Spectrofluorometer with single cell resolution. An innovative yet specific method of labelling bacteria is using phage conjugated with Quantum Dots. This method can be used for recognising both slow growing and highly infectious bacteria. This method was studied for detection of Escherichia coli using biotinylated phage and streptavidin coated Quantum Dots [33]. Quantum Dots coated with zinc [II]-dipicolylamine could detect only mutant and pathogenic Escherichia coli that lacks O-antigen and could also facilitate in vivo optical imaging of the infection in the animal [34]. It is also possible to identify the bacterial species by detecting the species specific DNA sequence. In this method of detection the Quantum Dots are made to bind to a probe which is complementary to the species specific DNA sequence. In a study this method was used for detection of Mycobacterium tuberculosis and Mycobacterium avium subsp. paratuberculosis where the biotinylated probe hybridized with the DNA was detected by streptavidin coated Quantum Dots [35]. This method was 70 to 90% accurate compared to real time PCR and had a limit of detection of 12.5 ng of DNA in 20µl. This technique can be implied in diagnostic assay for rapid, specific and sensitive method of pathogen detection. In another study Quantum Dots based molecular beacons were used to detect the antibiotic resistant β-lactamase genes in pUC18 of Escherichia coli [36]. The mechanism of detection used in the study was a single step FISH, which gave an excellent signal on genes in the plasmid. There was an attempt to use ferrichrome conjugated to Quantum Dots for detection of bacteria with receptors for ferrous ions [37]. This method was successful in detecting Pseudomonas fluorescens having ferrous receptors.
Detection of bacterial food contaminants
Many bacterial infections are spread by contaminated food. Thus, detection of bacterial contaminants in food is important to monitor outbreaks of food borne infections. Three food-borne pathogens namely Salmonella typhimurium, Shigella flexneri and Escherichia coli were detected by antibodies against each bacteria which was conjugated to Quantum Dots of different emission wavelength [38]. The same method of antibody conjugated Quantum Dots were developed for specific detection of Staphylococcus aureus in food and environment [39]. The bacteria were detected under a fluorescent microscope and limit of detection was found to be 900 CFU/ml. Most of the contaminants are found in meats, thus Quantum Dots can be used to analyse contaminant in meat sample. In a recent study, Quantum Dots were used to detect pathogens like Escherichia coli and Salmonella in ground beef by conjugating Quantum Dots to specific antibodies against the pathogens [40]. In a study chicken carcass wash water was used as a sample for detecting Salmonella typhimurium contaminant in chicken meat [41]. Here contaminant was separated from sample with antibody conjugated magnetic beads and then reacted to biotin tagged secondary antibody which binds to streptavidin coated Quantum Dots. In this method the limit of detection was in order of 103 CFU/ml. Also there has been a report of use of indirect immunofluorescence coupled to Quantum Dots for labelling and detection specific bacterial serotype of pathogen Vibrio parahaemolyticus attached to small marine animals which are pathogen carriers [42]. This method can also be extended for detection of other Vibrio species like Vibrio cholerae. With the use of Quantum Dots in biosensors and microarrays, the size of the instrument have been reduced and also permitted for automation thus increasing rapidity and sensitivity of the test. These techniques have been used in assaying Salmonellae [43]. Nowadays array systems have been developed for rapid and sensitive detection of bacteria as indicated by a study where Escherichia coli O157:H7 was detected using Quantum Dots conjugated to antibody and the concept of sandwich ELISA [44]. The above method gave a limit of detection of below 10 CFU/ml. Escherichia coli was also detected using colistin-functionalised Quantum Dots which gave a limit of detecting as low as 28cells/ml [45] with a short analysis time of 15 min. excluding preparation and photoactivation time. Detection of bacteria in Oral bio films It is observed that many bacteria exhibit a symbiotic relationship between each other. These bacteria form biofilm which is a complex cluster of bacteria. These biofilms are found widely in our environment and also in our body like mouth etc. It is desirable to study symbiosis for evolutionary studies. Quantum Dots can be used for specific detection of bacteria in oral biofilms. This method has been used in a study where specific human oral bacteria namely Streptococcus gordonii DL1, Streptococcus mutans UA159 and Veillonella spp. strain R1 were detected in a biofilm in vivo and in vitro [46]. Here immunofluorescence method of imaging was used, and this method does not depend on detection of bacteria based on their morphology, which makes this rapid technique amenable to automation, enabling the detection from numerous samples. These studies also give an insight on the spatial relationship between bacteria and interspecies interaction in biofilm. Presence of specific gene in the biofilm can give a large insight on the species of the bacteria and also function of that gene in the biofilm. In a report Bacillus spoOA gene was analysed in a biofilm using DNA-Quantum Dots system [47]. The hybridization of target DNA to the probe was detected using flowcytometer and had a limit of detection of 0.02 nM. It has been observed that biofilms made of Streptococcus sp. and Veillonella sp. are formed in early stage of the plaque. These results were confirmed by using Quantum Dot based immunofluorescence on the enamel surface [48].
TOXICITY OF QUANTUM DOTS TO BACTERIA
We have seen the brighter side of the Quantum Dots but it has been demonstrated that these nanoparticles are toxic to living cells including bacteria. Formation of reactive oxygen species by the interaction of Quantum Dots with other bio molecules or release of the heavy metal ions which constitute the Quantum Dots have been the mechanism of inducing toxicity to the cells. The toxicity of Quantum Dots on four different strains of bacteria namely Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Escherichia coli were studied [49]. The result indicated that Gram-positive strains were more resistant to the toxicity than the Gram-negative bacteria. This might be due to large amount of peptidoglycan on cell wall of Gram-positive bacteria. The study also indicated that Gram-positive bacteria previously exposed to Quantum Dot showed lack of reproducibility.
CONCLUSION
Today, the major hindrance for using Quantum Dots in bacteriological applications is its toxicity. It is suggested that by eliminating the heavy elements from Quantum Dots, the toxicity can be reduced. Thus new Heavy Metal-free Quantum Dots like carbon quantum Dots are being produced [50]. But their effective usage is still under research. The cost of production of Quantum Dots is also topic of concern when they have to be mass produced and used as diagnosis & analysing tools. Many methods for synthesizing biocompatible Quantum Dots using bacterial cells like Escherichia coli have been proposed [51]. The efficacy of these Quantum Dots in various applications will have to be proved. In conclusion, it can be said that Quantum Dot based techniques are rapid, sensitive and reliable tools for the enumeration, detection and differentiation of Bacteria.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesEFFECT OF EARLY DEFOLIATION ON THE NODULATION AND SOME AGRONOMIC TRAITS OF SOME SELECTED COWPEA (VIGNA UNGUICULATA L. WALP)
English95102Ogundele O. E. and Awosanya A. O.EnglishThis study investigated the effect of defoliation at emergence of cotyledons, 2 weeks and 4 weeks after planting on the number of nodules, nodule diameter, total nodule weight, nodule efficiency, root length and plant height of two improved varieties and one local variety of Vigna unguiculata (L.) Walp. efoliation treatments were carried out in triplicates using the CRD. Plants were raised in pots and data collected after 6 weeks. This study simulated early defoliation of the crop as evidenced in pest attack, disease infection and/or grazinganim als and significantly reduced nodule number (PEnglishagronomic, defoliation, cotyledons, nodulation, Vigna unguiculataINTRODUCTION
The problem of maintaining soil fertility in time past had led to the use of manure and fertilizers. However, these methods were constrained by erosion which leads to the washing away of the nutrients in the top soil. Alternative methods have since been used by planting legumes such as cowpea to maintain soil fertility and at the same time prevent soil erosion. Crop farmers now focus on legumes for the aforementioned benefits they attract and their ability to form association with bacteria that can fix nitrogen to the soil. Consequently, productivity is improved in terms of quality and quantity. Cowpea, a staple food with high protein, mineral and vitamin content is also valued for improving soil fertility through the formation of root nodules for nitrogen fixation. It plays an important role in agriculture by providing soil nitrogen. The symbiosis can help relieve the requirement for added nitrogenous fertilizer of other crops that are intercropped with cowpea. The legume is known for its symbiotic association with Rhizobium bacteria enabling it to fix atmospheric nitrogen (Laity et al., 2003). Bacteria of the genus Rhizobium play a very important part in agriculture by inducing nitrogen-fixing nodules on the roots of legumes such as cowpea. However, the growth of cowpea is limited by numerous biotic (diseases, insects and other pests) and abiotic factors that can cause serious devastation (Rahman et al., 2008). Several foliage defoliators, including insect pests have been reported to cause severe defoliation of cowpea. Aphids (Aphis craccivora) attack cowpea especially at the early vegetative stage (IITA, 2000) while a considerable number of insects belonging to Acrididae and Lepidopteran larvae have been reported feeding on cowpea leaves, “skeletonizing” and sometimes defoliating the entire plant. Other foliage defoliators of cowpea belong to the family Chrysomelidae (Allen et al., 1996). Several studies in crop defoliation have received attention in the last four decades using various crops. Studies have also been carried out to determine the effects of defoliation at the different growth stages on yields of crops, as it simulates pests and disease damages. Some studies (DeBeer, 1983; Schneiter et al.,1987; Schneiter and Johnson, 1994) described an interaction between the severity of damage (percentage defoliation) and the growth stage at which this occur. Some of the earlier studies on defoliation of cowpea were particular about defoliation at the different growth stages (Rahman et al., 2008) and situations peculiar to America and Europe. There is probably little or no information on cowpea defoliation at the vegetative stage in Nigeria where the crop is largely grown. Defoliation as it may be caused by pests and disease conditions affects the growth of the entire plant. Perhaps nodulation of the crop is also affected by the defoliation thereby having a great impact on nitrogen fixation and consequently crop productivity. This study was therefore conducted to determine the effects of defoliation at the vegetative stage of growth on the nodulation of some selected varieties of cowpea Vigna unguiculata (L.) Walp. in the South-Western part of Nigeria.
MATERIALS AND METHODS
Cowpea Varieties
Two improved varieties of Vigna unguiculata (L.) Walp. (IT89KD-288 and Damila) from IITA and a local variety (Mata) in Nigeria were used for the research.
Preparation of Experimental Pots
The experiment was a completely randomised design with 36 experimental pots arranged in greenhouse. The experiment was carried out at the botanical garden of the biological sciences department in Tai Solarin University of Education, Ijagun, Nigeria between March and April. The experimental pots were perforated at the base to allow for proper drainage. Loamy soil from the botanical garden was used.
Planting
Three hand-picked healthy seeds were planted in each pot and later thinned to one after germination. Cultural practices such as weed control were also carried out regularly.
Treatments
There were 4 treatments including the control for each variety. The treatments were:
Defoliation immediately after emergence of cotyledons;
Defoliation 2 weeks after planting;
Defoliation 4 weeks after planting;
No defoliation (control) Each treatment was done in triplicates.
Data Collection
Data were collected 6 weeks after planting. In determining the number of nodules, each pot was carefully emptied and the soil on the root was gently washed with water. Root nodules were physically counted with the aid of hand lens. The nodules present on the roots of each plant were collected and weighed with electric weighing balance to obtain the total nodule weight. Nodule diameter was measured with the aid of a micrometer screw gauge. Nodule efficiency was determined by examining the root nodules of each plant to check for the presence of the reddish-pink pigment (leghaemoglobin), which indicates a functional or efficient nodule. Plant height and root length were measured using meter rule. Dry mass of shoot was measured using an electric weighing balance after shoots had been oven-dried at 60OC for 72hours.
Data Analysis
Analysis of data was done using ANOVA with Genstat® statistical package.
RESULTS
The effect of defoliation on the number of nodules was highly significant in the three varieties used (PEnglishhttp://ijcrr.com/abstract.php?article_id=2308http://ijcrr.com/article_html.php?did=23081. Allen D J, Ampofo J K O, Wortman C S (1996). Pests, diseases and Nutritional disorders of the common bean in Africa. A field guide. The Netherlands, CTA, 132pp.
2. Anonymous 2009. Rhizobium and Legume Root Nodulation.Http://www.microbiologyprocedur e.com/rhizobium-and-legume-rootnodulation/temperature-and-light.html
3. DeBeer J P (1983) Hail damage simulation by leaf area removal at different growth stage of sunflower. Gewasproduksie. 12: 110-112.
4. Hartwig U A and Nosberger J (1994). Plant and soil 16(1): 109-114.
5. IITA, 2000. Crops and farming systems. Http://www.iita.org/crop/cowpea.htm.
6. Laity F, Diouf D and Fall M A (2003). Genetic diversity in cowpea varieties
7. determined by ARA and RAPD techniques. Afr. J. Biotechnol. 2: 48-50.
8. Mondal M H, Brun W A, Brenner M L (1978). Effect of sink removal on photosynthesis and senescence in leaves of soya beans plants. Plant physiology. 61: 394-397.
9. Muro J, Irigoyen I, Militino A F, Lamsfus C (2001). Defoliation effects on sunflower yield reduction. Agronomy Journal. 93: 634-637.
10. Rahman S A, Ibrahim U and Ajayi F A (2008). Effect of defoliation at different growth stages on yield and profitability of cowpea (Vigna unguiculata L. Walp.) E. J. Env. Agric. & Food Chem. 7(9): 3248-325.
11. Schneiter A A, Johnson B L (1994). Response of sunflower plants of physical injury. Can. J. Plant Science. 74: 763-766.
12. Schneiter A A, Jones J M, Hammond J J (1987). Simulated hail research in sunflower defoliation. Agronomy Journal. 79: 431-434.
13. Selter T L, Brun W A, Brenner M L (1980). Effect of obstructed translocation of leaf abiscic acid and associated stomatal closure and photosynthesis decline. Plant physiol. 65: 1111-1115.
14. Shibbles R, Secor J, Ford D M (1987). Carbon assimilation and metabolism.Agronomy. 16: 535-588.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareINTERESTING FACTS ON THYMUS GLAND CHANGES-A REVIEW
English103109Praful S. JevoorEnglish MathadaV. RavishankarEnglish Sandhya DharwadkarEnglish Amit MagadumEnglish Somashekhar BiradarEnglishIn most of the cadavers procured for the dissections in the department of anatomy belongs to male adult cadavers, which were devoid of active thymus gland and often the gross features of this gland was ignored because of its remnants were found with variable degree of shape and sizes. During dissections in most of the cadavers we have often noticed the morphology of most thymus remnant from simple flat mass to a well defined solid mass of thymic tissue. To test our curiosity on microscopic studies we have noticed features like, abundant number of iscrete lymphocytes along with Hassall?s corpuscles were appreciated under hematoxylin and eosin staining. The thymus being a central lymphoid organ develops from endoderm of third pharyngeal pouch along with the parathyroid gland. It remains active till the age of 14 years and later shows gradual inclination towards its involution as conventionally it is believed to be an involuting organ with advancing age. Thymus being an important immune competent organ which has created a curiosity regarding its changes from advancing age to number of invasive and non invasive experimental studies, such interesting facts were considered to drag the attention of involuting organ.
EnglishAcetylcholine, Adipocytes, Hassall?s corpuscles, ImmunosenescenceINTRODUCTION
Thymus is a central lymphoid organ located in the superior mediastinum; many times it was ignored during the dissections of adult cadavers practical point of view its morphology was less stressed as it is replaced by the fatty tissue in most of the adult cadavers. Immunologically it is considered to be an important gland which programs the stem cells which are originating from bone marrow for its specific antigen activity. Regarding the involution of the thymus gland and its significance is sited in the texts of histology, this organ created curiosity in connection with its versatile changes in different age, sex and disease conditions, such as immune deficient ailments varying from opportunistic susceptibility of body to simple viral infections to life threatening condition like acquired immunodeficiency syndrome(AIDS). The thymus arises from the third pharyngeal pouch of thymic primordia at the end of the fourth week in the form of endodermal proliferations, these endodermal proliferation forms a hallow tubes that invade the underlying mesoderm which later transforms into solid branching cords, these cords are the primordia of the polyhedral thymic lobules. The whorl like Hassall?s corpuscles within the medulla apparently arises from the ectodermal cells of the third pharyngeal cleft shortly after the formation of thymus is subsequently infiltrated by lymphocytes. As the thymus is developed along with many structures in the primitive pharyngeal floor during the fetal life, often it tend to take a different pathway and destination with or without influencing its accompanied structures 1,2 . Such rare incidence of embedded thymic follicles within the section of thyroid gland was accidentally found during the histopathological investigations showing its diverse destination3 . The congenital anomaly like digeorge syndrome is often associated with maldevelopment of thymus gland. The thymus is highly active during the perinatal period and continues to grow throughout childhood reaching its maximum size at puberty and later on the gland involutes rapidly which is replaced subsequently with fatty tissue4 . The importance of the thymus often reviewed in adults with the suspected clinical locomotors functions, most of such signs and symptoms are often proved with persistent activity of thymus. In case of adults the thymus often start producing its antigens against own body components which can lead to condition like myasthenia gravis this condition can be handled effectively by thymectomy 5 . During routine dissections in most of the cadaveric gross thymic remnant was noticed as a mass resembling the original gland as shown in the photo legend (1) and its microscopic appearance has shown in the photo legend (2).
DISCUSSION
If we observe the whole life span of an individual as the age advances some of the bodily organs are showing obvious changes from simple atrophy to involution, often these changes are so subtle that the functional derangements are correlated with advanced structural changes in number of organs. During the early period of post natal life thymus gland plays a significant role of programming the cells which are reaching the gland for their charged specific tasks, such cells are destined to be involved in immune mechanism, our body defense activities are influenced by factors like age, sex, environment, food, nutritional status, genetic factor, hormonal status etc. During the dissections in most of the male cadavers often we have noticed and collected a lobulated mass situated at the upper and anterior part of the pericardium in the region of superior mediastinum(Photo legend-1), where usual thymus gland is situated. Such mass was studied thoroughly from out side and was later subjected to histological tissue processing, embedding, microtome sectioning and it was stained with hematoxylin and eosin, later on microscopic analysis it has shown a classical involuntary changes in most of the areas which resembles the thymic tissue and the Hassall?s corpuscles (Photo legend-2) were sparsely spread among dense fatty tissue along with large number of scattered lymphocytes. These findings have confirmed that it is an involuted thymic remnant. Thymus gland shows a linear increase in its weight as the age advances till the 16 years of age, later it undergoes gradual loss in its gross weight 6 . Thymus is being a primary lymphoid organ plays an important role in the development of other secondary lymphoid organs. This fact can be realized by the non-development or immature development of other secondary lymphoid organs followed by thymectomy during the early postnatal life, such dependent activity makes one to become more susceptible to infections from simple to complex nature. The activity of thymus begins in the early fetal life itself as a source of production of diversified T cell production which is further boosted in the early stages of post natal life, but its function declines as the age advances. The neonatal thymectomy results in reduced number of T cell counts in short and long term complications which shows its indispensable functional significance through infection prone attitude in infants with advancing ageing process. 7 The developmental anomalies like digeorge syndrome often associated with the absence of thymus gland, during such situations clinical trials associated with the transplantation of the thymus gland in early childhood was found with formation of new T-cells after a span of 5 months suggesting immune reconstitution which was able to render an essential support for host to fight against the entry of foreign bodies 8,9.It is commonly seen that the advanced ageing process affects all aspects of immune system followed by thymectomy during infancy a particular type of “ T “cell activity in an individual is often characterized by lower CD4 + and CD8+ cell number and diversity. The removal of thymus in infancy results in premature onset of decline in immune competency, often such individuals are susceptible for infectious diseases, such clinical findings shows premature signs of immune aging and Immunosenescence which was experimentally correlated with the consequence of thymectomy in the early postnatal period 10. There are some experimental results correlating that the early thymectomy in case of the female rats are showing the precautious sexual maturity but the same procedure in case of the male rats showing no significant effects 11. A prospective randomized study treatment with growth hormone in HIV infected adults were showing changes in the body in the form of increased thymic mass along with increase T cell receptor rearrangement, such studies are indicating that the growth hormone is an important factor which can regulate the T cell development which can reverses the adverse influence on thymopoiesis in rodents. The thymus being a primary site of de novo cell production it will modulate the recovery in immune deficient cells, which supports the hypothesis of enhancement of thymus activity by growth hormone administration in immune deficient conditions. The possibility of the functional reinstate of the thymus was observed in a condition where the growth hormone treatment can revive the thymic function by enhancing the thymopoiesis 12. Such immune reconstitution studies can support the highly virulent HIV infections which are targeting the thymus where it shows a strong correlation associated with reversible changes in the remnants of thymus gland 13, 14 .The age factor of an individual and the capacity of thymus reversibility are seems to be interdependent one where the immune modification process shows difference in childhood and adulthood 15 . A relatively common clinical condition where the derangement in functional activity of the primary lymphoid organ due to autoimmune factors targets the normal neuromuscular mechanisms leading to condition like myasthenia gravis where the surgical evidence of thymectomy and its clinical impact on the prognosis of the myasthenia patients have shown that the thymus is profoundly involved in pathogenesis by synthesizing autoantibodies against acetylcholine (Ach) receptors. In more than 70% of all the myasthenics the thymus gland shows lymphofollicular hyperplasia, where the thymectomy appears to be one of the most rewarding therapeutic measure16 . Though the thymus is showing its involution process in either sex, indeed as the age advances the gland is obviously showing some discrimination. These changes could be due to different hormones involved in driving the advance age related changes in an individual. Stage like pregnancy has overall influence on the body of an individual which was studied in mammals has shown that the thymus loses its weight and cellularity with a marked involuntary changes in the cortical portion. The increase in the levels of hormones in pregnancy is generally believed to be one of the causative factors for its altered morphology and function. The thymus cortical part is the site of the cell trafficking in and out of the thymus, the small MER (Medulla termed epithelial medullary ring) seen in virgin mice with the advancing age, where the MER will enlarge and much more expansion is seen in its size during pregnancy. In the later stages of pregnancy the cortical epithelial cells appears too effete and most of the macrophages become heavily laden with apoptotic cells followed by drastic changes in the thymic microcirculation which could affect the secretion of cytokines and other products of the thymus 17 . The collection of thymus in diversified age group of people and when they were subjected to micro anatomical studies have found that during the biopsies obtained from the patients immediately prior to partial thyroidectomy for nontoxic nodular goiter, on its microscopic examination 50% of the glandular cell activity declination was recognized especially in late adolescent individuals and 10% in case of late middle aged individuals. In men the age involution is linear whereas in women it is biphasic. In humans during second and third decades the proportion of the gland parenchyma is relatively less in women when compared to men. But with advanced ageing process there is a little sex difference where the medulla shows a continuous linear involution which is similar in both the sex. But with respect to cortical part of the thymus in case of men which has shown accelerated atrophy in contrast with the changes in age matched women. Probably
the sex related changes in the thymus attributed to hormonal levels in the body which were experimentally correlated 18 . The number of invasive and investigative studies were showing some interesting structural changes in many organs with the help of improved scanning techniques where the drastic revolution in the field of radio diagnostics has helped us to understand and correlate the structural and functional changes of many more organs19,20 .
CONCLUSION
Thymic parenchyma atrophies with advancing age and it is replaced by fatty tissue it may be diffuse or focal. The thymic remnants are showing wide changes from infancy to adulthood, in different sex, stressful and disease status of the body. Unlike our conventional thoughts its capability of functional and structural reversibility in immune deficient adults certainly taps possibility of its functional revival, where the thymus gland is showing its versatile morphological and histological pattern in the body is associated with a wide range of immune mediated response. Indeed such peculiar experimental out come makes one to go further in the direction of the research related to immune modulation where the thymus stands as a peculiar and interesting organ.
COMPETING INTERESTS-
Here the authors declares that they don?t have any competing interests.
ACKNOWLEDGEMENT-
The author would like to acknowledge the support provided by Dept. of Anatomy, JN medical college, KLE University, Belgaum.
FUNDING-
This work has not utilized any funding or financial aid from any of the sources.
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4) Edwin et al. Age related changes in the cellular composition of the thymus in children. The journal of allergy and clinical immunology. 2005; 115(4):834-840.
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11) Jr. Ahearn T F. "The Relation of the Thymus Gland to Sexual Maturity" http://ecommons.luc.edu/luc_theses/435 ( date of access 26-4-2012)
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareINDIGENOUS UNCOATED AND HYDROXYAPATITE COATED COMMERCIALLY PURE TITANIUM FOILS FOR GUIDED BONE REGENERATION IN DEFECT SITES OF IMPLANTS - AN IN VITRO STUDY
English110121Ranzani. R.English Abby AbrahamEnglish Chakravarthy.R.English Lakshmi.S.EnglishUse of barrier membranes for the regeneration of bone defects has significantly changed implant dentistry in the past
years. The design of the membranes employed in this study plays an important role in establishing a healing
environment by separating tissues during healing and thus providing space for regeneration of bone in cases of
insufficient bone around implants, it acts like a tent in cases of extraction sites- which is a potential site for future
implant placement thus reducing the problems of irregular ridge defect, and the rigidity of the material prevents
tissue ward collapse of the membrane.
The present study was conducted to prepare the titanium in the form of foil, and tested for biocompatibility using
fibroblast cells, hydroxyapatite coating was made on the foil, characterization by X-ray diffraction for both the
uncoated and the coated foils was done, and osteoblasts adhesion test was done and viewed under scanning electron
microscope both before and after the test.
Materials and methods: Commercially pure titanium foils were made using 16% hydrofluoric acid. 3×3 cm sheets
were cut and dipped in the acid intermittently and alternatively dipped in water and wash the acid. For every minute
of etching the material is measured using a micrometer to check the thickness of the foil. It is carried on until it is
60μ thick and uniform pores appear. Hydroxyapatite coating was done over the foil by dip coating in a solution
made by a mixture of hydroxyapatite powder, water, glycerine, polyethylene glycol, and ethanol. The foil is dipped
and withdrawn at 45? a speed of 10mm/ 15 seconds. Then the foil is dried in an oven at 100 ? and heat treated in a
microwave oven for 45 minutes.
Results: The titanium foil was treated for biocompatibility using fibroblast cells, and the test showed that the
material was biocompatible. After coating with hydroxyapatite, the uncoated and hydroxyapatite-coated titanium
foils and the hydroxyapatite powder were characterized by x-ray diffraction and the dip coated foils clearly showed
the hydroxyapatite peaks along with the substrate titanium peaks. Scanning electron micrographs of the uncoated
foil revealed uniform distribution of through and through pores and scaly appearance in between pores. The coated
foil revealed uniform distribution of hydroxyapatite coating. And the thickness of the hydroxyapatite in cross section
was 50μ. Osteoblasts adhesion test was conducted on both the uncoated and the coated foils and scanning electron
microscopic study was conducted. The micrographs revealed the adhered osteoblasts on the surface of both the
uncoated and the coated test samples.
Conclusion: A new membrane material titanium foil was prepared by acid etching, and the material proved
biocompatible in fibroblast culture study. The foil was dip coated with hydroxyapatite. X-ray diffraction showed
foils with hydroxyapatite peaks along with the substrate titanium peaks. Scanning electron microscopic study
revealed micro pores and scaly appearance of the uncoated foil, and for the coated foil it revealed a uniform coating
of the hydroxyapatite. The surface roughness of the foil has given provision for the attachment of the
INDIGENOUS UNCOATED AND HYDROXYAPATITE COATED
COMMERCIALLY PURE TITANIUM FOILS FOR GUIDED BONE
REGENERATION IN DEFECT SITES OF IMPLANTS – AN IN
VITRO STUDY
Ranzani. R., Abby Abraham, Chakravarthy.R., Lakshmi.S.,
Department of Prosthodontics, Meenakshi Ammal Dental College,Chennai
–600 095, India
E-mail of Corresponding Author: drchakra19@gmail.com
110 International Journal of Current Research and Review www.ijcrr.com
Vol. 04 issue 17 September 2012
hydroxyapatite. Osteoblasts adhesion test was done and scanning electron microscopic study was conducted to view
the adhered cells. The osteoblasts cells have adhered to the substrate of both the uncoated and the coated test
samples. The results revealed that this study could give rise to a new generation of osseo conductive membranes for
use in implant defect sites.
EnglishTitanium foils, Hydroxyapatite coated foils, Fibroblast culture, Osteoblast adhesion test, Scanning electron micrographs. Introduction “Everything has its time” The dinosaurs had their time, and so will it once be said that amalgam had its time too.In dentistry, implantology currently is in “its time”. Other than the discovery of osseointegration more than twenty years ago, the concept of guided bone regeneration represents the most important progress in implant dentistry. This material helps in following cases: 1. In cases of insufficient bone around implants, 2. Acts like a tent over the graft material, 3. Preserves the clot formed by acting as a tent in cases of extraction site, which is a potential site for future implant placement. 4. The rigidity of the material prevents tissue ward collapse of the membrane. The present study was conducted to obtain an improved uncoated and hydroxyapatite coated titanium foils, both of which can be an economic replacement for their costly counterparts. Aim of the study: 1. To prepare the titanium in the form of foil. 2. To test biocompatibility of the material using fibroblast cells. 3. To make hydroxyapatite coating on the foil. 4. X-ray diffraction done to characterize the surface of both the uncoated and hydroxyapatite coated foils. 5. Scanning electron microscopic study done to evaluate the surface of both the uncoated and hydroxyapatite coated foils. 6. Osteoblasts adhesion test done to evaluate the effectiveness of the materials in osteoblasts adhesion which would potentiate the growth of bone around implants. 7. Scanning electron microscopic study following osteoblastic adhesion test. Materials and methods: 1. Commercially pure titanium sheets- grade I (ASTM) (Midhani grade – Titan 12), 99.8% - Titanium, 0.2% - Iron, 0.1% - Oxygen, 0.05% - Nitrogen. 2. 16% Hydrofluoric acid 3. Glass beakers 4. Glass measuring jar 5. Pipettes 6. Micrometer 7. Acetone 8. Ethyl alcohol 9. Distilled water 10. Hydroxyapatite powder - 7.3% 11. Ethanol (99% pure) 12. Polyethylene glycol 600 13. Glycerol. Equipments: 14. Ultrasonic cleaner, 15. Autoclave, 16. BPL micro convection system, BMC-900 T, 17. X-ray diffraction unit, 18. Scanning electron microscope, XL30 SEM, PHILIPS, 19. Splutter coater- Hitachi. Methods: 1. Preparation of titanium foil (fig: 1 a, 2a,2b,2c) The titanium sheets were cut to a dimension of 3cm×3cm. 16% hydrofluoric acid w/v was taken in a glass beaker, and the cut sheets are dipped in the acid intermittently, and it is alternatively dipped in water to wash the acid. For every minute of etching, the material is measured with a micrometer to check the thickness of the foil. This procedure is carried on until the material is 60μ thick and uniform pores appear. 2. Ultrasonic cleaning of the foil: Foils are ultrasonically cleaned in a) Acetone for 20 min, b) 70% ethyl alcohol solution for 20 min and c) Finally in distilled water for 20 min. 3. Preparation of hydroxyapatite coating over the titanium foil (fig:1b, 2a, 2b, 2c): The ingredients of the dip-coating solution and their weight percentages are: a) Hydroxyapatite – 7.3% b) Ethanol – 66.2% c) Polyethyleneglycol 600 – 2.2% d) Glycerol 10.2% e) Distilled water – 14.1% Preparation of dip coating solution (flow chart no:1): 1. A mixture of hydroxylapatite and water is made and maintained for 8 minutes. 2. When this mixture is around 2 minutes, polyethylene glycol and ethanol are mixed in a separate beaker and maintained for 8 minutes. 3. Glycerol is added to the hydroxyapatite and water mixture and maintained for 2 minutes. 4. After this time the polyethylene glycol and ethanol mix is added to the mixture of hydroxyapatite, water and glycerol. 5. Then the whole solution is maintained for 4 minutes. The final solution is the dip-coating solution. Titanium foil is dipped and withdrawn at 45 degree angle in a freshly prepared dip-coating solution at a constant speed of 10mm/15 seconds. This procedure was repeated 2-3 times to increase the coating thickness.The coated foils were immediately dried in an oven for 45 minutes. The microwave oven used in this process is a BPL micro convection system. 4. X-ray diffraction (fig 3a & 3b): The hydroxyapatite powder, titanium foil, and hydroxyapatite-coated foil were characterised by x-ray diffraction technique. The dip-coated titanium foils clearly shows the hydroxyapatite peaks along with the substrate titanium peaks. Fibroblast culture Materials and methods: Materials : 1) Commercially pure titanium foils, 2) Distilled water, 3) Glaxo modification eagle?s medium (GMEM) 4) Cell line: VERO cells. 5) Fetal calf serum were procured from HI media, India and stored at -20 degree Celsius. 6) Antibiotic stock: Antibiotic stock was prepared with the following antibiotics in distilled water and added to the medium in the following concentration. Gentamycin : 50 mg/lit Penicillin : 1,00,000 units/litre Streptomycin: 50-100 mg/litre Fungizone : 25μ / litre 7) Sodium bicarbonate stock: Sodium bicarbonate 8.8% solution (w/v) in phosphate buffered solution was autoclaved and stored at +4 degree Celsius until required. 8) Trypsin versene glucose:
Trypsin : 0.5 gm Glucose : 0.2gm EDTA : 10.2 gm PBS : 200ml Trypsin versene glucose was sterilized by filtration through 0.22 μ membrane filters, ale quoted and stored at -20 degree Celsius until required. 9) Phosphate buffered saline 10X Sodium chloride : 80.0gm Potassium chloride : 2.0 gm Sodium hydrogen Orthophosphate : 11.33 gm Potassium di hydrogen Orthophosphate : 2 gm Distilled water : 1000 ml. Phosphate buffered saline (10X) was prepared in sterile distilled water from this 1 litre of 1X was prepared and sterilized by autoclaving at 121 degree Celsius for 15 minutes at 15 lbs pressure. Instruments: 1) Plastic culture plates: 6 well tissue culture plates (NUNC). 25 sq.cm and 75 sq.cm tissue culture bottles (Greiner labor technik) were prepared for propagation of the cells. 2) Glass wares: glass beakers, Glass measuring jar, Micro pippetes, Test tubes, All glasswares were of borosil. Equipments: 1. Ultrasonic cleaner 2. Light microscope with camera attached 3. Incubator. Procedure: 1. Sterilization of glassware?s: The glassware?s were washed thoroughly, with mild detergents and rinsed thoroughly with distilled water, dried and covered with aluminium foil and sterilized in autoclave at 121 degree Celsius for 20 minutes at 15 lbs pressure or in hot air oven at 180 degree Celsius for 2 hours. 2. Preparation and sterilization of medium: The powdered medium was dissolved in 960 ml of distilled water, tryptose phosphate broth (10%) was added and the pH was adjusted to 7.4 with sodium bicarbonate and 2.3 ml/L of antibiotic stock was added and sterilized by filtration through 0.22μ membrane filter and stored at+4 degree Celsius. 3. Maintenance of VERO cells: VERO cells were maintained in growth medium (GMEM 8% FCS and antibiotics). VERO cells were passaged at a split ratio of 1:3 on every 3- day. Briefly spent medium from bottles (25 sp cm), containing confluent monolayer were decanted and the cell layer washed with trypsin versene solution twice. Detached cells were harvested in 3ml of growth medium and resuspended to make 24 ml of cell suspension in growth medium. Cell suspension was seeded into 3 bottles (25-sq cm) at the rate of 8 ml per bottle and incubated at 37 degree Celsius. 4. Extraction: The extraction liquid resulted from the incubation at 37 degree Celsius for 120 hours of the material in the extraction “vehicle” (minimum essential medium) under specific conditions. A blank extraction was done using Glaxo modification earl?s medium under the same conditions, except for the absence of material. The cells were tested by direct contact of extraction liquid of the test implant material. 5. VERO Cyto toxicity study: This was performed in 6 well tissue culture plates (NUNC). Cells were seeded at the rate of 2×105 cell/ml. 1.5ml of trypsinised cells were added with equal volume of each of the four metal extract in growth medium. 1.5ml of this mixture was seeded into one well of the plate. Four such extracts and negative controls were tested in a single plate. Cytopathic effect was observed daily for four days. At the end of which the cells were stained with Giemsa. Result of the test (fig 4a & 4b): VERO cyto toxicity: none of the metallic extracts caused any cytopathic changes in the VERO cells, when the extracts were used as neat. Osteoblasts adhesion test Materials: 1. Disodium phosphate, 2. Sodium dihydrogen phosphate, 3. Phosphate buffer (0.1 M, pH=7), 4. Glutaraldehyde : Make up 2.5 ml 25% glutaraldehyde to 25 ml using phosphate buffer, 5. Isopropyl alcohol , 6. Isoamyl acetate, 7. Trypsin, Instruments: 1. Multiwell plates, 2. Glass beakers, 3. Pipettes, 4. Glass measuring jar, Equipments: 1. Laminar flow bench, 2. Carbon di-oxide incubator, 3. Inverted phase contrast microscope, 4. Scanning electron microscope.
Procedure:
Cell culture system:
Osteoblasts were maintained with minimum essential media supplemented with foetal calf serum.
Maintenance of cells:
1. When the cells become confluent, subculture the cells using trypsin.
2. Incubate until the cells become rounded and begin to detach by checking under microscope every one minute.
3. Add 4ml of serum containing medium and disperse the cells carefully using a Pasteur pipette.
Adhesion test:
1. Test samples were conditioned with media.
2. Cells were seeded on to the conditioned test samples, standardized to 1.5 x 1.5cm, in multiwell plates with required amount media.
3. Incubate the culture for 24 hours.
4. After 24 hours, discard the medium from the culture dish.
5. Rinse the cells with phosphate buffered saline.
6. Fix in 2.5% glutaraldehyde solution in phosphate buffer for three times.
7. Dehydrate in different grades of alcohol.
8. Immerse in asoamyl acetate for 2 minutes.
9. Then processing is done for critical point drying, gold coating and observation under scanning electron microscopy.
Result of the test (fig: 5a,5b,5c, 6a, 6b, 6c ):
Osteoblasts adhered on to both coated and uncoated test samples. Cell covered surface is seen in scanning electron micrographs of 400x. At higher magnification spreading of an individual cell can be seen.
RESULTS
The result in the present study indicates that a possible new way of thinking about implant placement may be on the horizon. The foil prepared was 60μ thick with uniform distribution of pores, which can act as a passage for the nutrients to reach the underlying tissues. Four samples of 15mm X 15mm dimension were subjected to fibroblast culture study. Extract of the material was prepared and tested for cytotoxicity. None of the metallic extracts caused any cytopathic changes in VERO cells when observed for four days. Thus proving the biocompatibility of the material. The hydroxyapatite coating was done by preparing a dip coating solution. When assessed by X-ray diffraction the coating material was identified as hydroxyapatite and the substrate as titanium. Scanning electron microscopic study revealed that the uncoated material had distribution of pores that were through and through, the area in between the pores appeared scaly and rough, proving the ability of the material to retain the hydroxyapatite coating. In the coated material, hydroxyapatite was uniformly distributed on the substrate. The vertical section showed that the thickness of the hydroxyapatite material was 50μ. Osteoblast adhesion test was performed for both coated and uncoated test samples. 1.5 x 1.5 cm test samples were conditioned with media and cells were seeded on the samples in multiwell plates. Step wise procedures were conducted and observed under scanning electron microscope. Micrographs showed that the osteoblasts had adhered onto both the coated and the uncoated test samples. Cell covered surface was seen, and at higher magnification and spreading of an individual cell was seen.
DISCUSSION
The term guided bone regeneration, has been used in tissue engineering for some years and is actually a specialized sub-area of tissue engineering and using this procedure for treatment of partial and total edentulism with dental implants has become an accepted treatment modality. Guided bone regeneration is based on the concept that most tissues are capable of selfreconstitution if appropriate conditions exist and by compartmentalizing wound healing. By placing a physical barrier between epithelial and connective tissues on one side and implants and bone on the other side, guided bone regeneration procedures aim to create a protected space for the blood clot to form and organise. The presence of a cell occlusive membrane is required to prevent the migration of epithelial and connective tissue cells into the wound area, thus allowing bone cells to form from marrow spaces to repopulate the defect and to mature into new bone. Membranes used in guided bone regeneration should possibly achieve a good degree of tissue integration with neighbouring connective tissues in order to obtain a mechanically stable environment necessary for successful bone and soft tissue healing. However regeneration of the bone tissue may be influenced by the nature of the membranes themselves and regeneration takes place in sequence of events including blood clot formation, angiogenesis, osteoprogenitor cell migration, woven bone formation, compaction of woven bone and secondary remodelling. In this study, we have made indigenous commercially pure titanium foils by etching, until it is 60μ thick and uniform pores are seen. Hydrofluoric acid is used because it attacks titanium even at very dilute concentrations. Hydroxyapatite coating is done on the foil by dip-coating, in a solution prepared by a combination of hydroxyapatite powder, ethanol, polyethyleneglycol, glycerol, and distilled water. Hydroxyapatite is used because of its osteoconductive and osteophilic properties.
Assessment of fibroblast culture:
The titanium was tested for biocompatibility using fibroblast cells. Metal extract was prepared by incubation at 37? C for 120 hours of the material in the extraction vehicle. Trypsinised VERO cells were added with equal volume of the metal extract in the growth medium. 1.5 ml of this mixture is added to 5 wells of the six well plate and negative control were tested. The cells were observed daily for four days. None of the metal extract caused any cytopathic changes, thus proving the biocompatibility of the material. Assessment of x-ray diffraction: The hydroxyapatite powder, titanium foil and hydroxyapatite-coated foil were characterized by x-ray diffraction. The uncoated foil clearly shows the titanium peaks and the coated foil shows the hydroxyapatite peaks along with the substrate titanium peaks.
Assessment of osteoblasts adhesion test:
Although the study of fibroblasts provides valuable information about its response to exogenous materials, cells of different origin react differently to foreign bodies. Thus, the results from fibroblasts should not be transferred to bone cells. The uncoated and the coated samples were tested for osteoblasts adhesion. Cells were seeded on to the surface of the materials. The material along with the medium was incubated for 24 hours. Later the medium was discarded from the culture dish and processed for critical point drying, gold coating and observed under scanning electron microscope. The surface chemistry of biomaterials can directly affect cell behaviour and interactions between the host environment and the biomaterials may have significant downstream consequences. Therefore the nature of the materials used in implantology may drastically influence surrounding bone regeneration. Cell attachment and migration are dependent on cell/ substrate interaction. The surface properties of the material have a strong biophysical influence on the cell kinetics. Osteoblasts are an anchorage – dependent cell type and need to attach and spread so as to divide and become confluent. The cytoskeletal response (actin filament and focal contact formation) of osteoblast-like cells to various substrates has been studied by many investigators. Bone cells are known to secrete a great amount of transforming growth factor-β (TGF - β1) and express cell surface TGF – β1 binding sites. Produced at all stages of bone remodelling, TGF – β1 is a potent mitogen in osteoblastenriched cultures from foetal tissue. It increases the expression of many genes associated with osteoblast activity and the production of extra cellular matrix macromolecules such as type I collagen, fibronectin and osteonectin. Hasegawa and coworkers1 demonstrated that the conformational state of proteins can influence cell activity and consequently extra cellular matrix formation. This might be one of the reasons why cells, because of more favourable spreading reaction, increased their rate of growth and supported the formation of mineralized matrix. The role of TGF-β1 in bone regeneration is mediated by changes in extra cellular matrix macromolecules, which, in turn, are regulated by a balanced secretion of biologically active growth factors. Human osteoblast synthesis of these growth factors and extra cellular matrix macromolecules may be influenced by the materials used in barrier membranes. Guided bone regeneration is a means of using the osteogenic potential of progenitor bone cells, to create new growth in a variety of osseous defects. Osteogenesis on bioactive substrates in characterized by a temporal sequence of biologic events involving cell morphology, proliferation and differentiation Hurzeler and associates2 found a substantial amount of “re-osseointegration” after treating ligature peri-implantitis using a combination of guided bone regeneration and bone grafts.
Assessment of scanning electron micrographs before osteoblasts adhesion:
The coated samples exhibited a thickness of 50μm of hydroxyapatite coating in cross section. When the surface was viewed, the hydroxyapatite crystals were uniformly distributed and uniform in size. The surface of uncoated foil revealed uniform distribution of pores and the surface in between the pores had a scaly appearance, which aided in both adhesion of osteoblasts and hydroxyapatite coating.
Assessment of scanning electron micrographs after osteoblasts adhesion:
The micrographs revealed that osteoblasts adhered to both the uncoated and the hydroxyapatite coated samples. Cell covered surfaces were seen in the micrographs of 400x. at higher magnification spreading of an individual cell was seen. Thus the material used in this study has proved to be osseoconductive by promoting the adhesion of osteoblasts to the substrate. The present study is an in vitro study. For further clinical application an in-depth invivo study is of paramount importance.
Summary
The study was conducted to prepare a titanium foil, which can be used as a membrane for guided bone regeneration in defect sites of implants. Titanium sheets of required dimensions were cut and immersed in hydrofluoric acid intermittently and measured every minute of etching using micrometer until the foil is 60μ thick and distribution of micropores is present. The material was subjected to fibroblast culture to evaluate the biocompatibility. The results indicated that the material is biocompatible. The prepared foils are dip-coated, and the uncoated and the coated foils were characterized by X-ray diffraction technique. The uncoated foils clearly shows the titanium peaks and the dip-coated titanium foils showed the hydroxyapatite peaks along with the substrate titanium peaks. Scanning electron microscopic study showed micropores in the uncoated material and the scaly substrate in between the pores. And the hydroxyapatite coated material showed uniform coating of the material. Osteoblasts adhesion test was conducted on both the uncoated and coated test samples. The scanning electron micrographs revealed the adhered osteoblasts on the surface of both the test samples.
CONCLUSION
1. A new membrane material, titanium foil was prepared.
2. The material was proved biocompatible in fibroblast culture study.
3. This material was dip-coated with hydroxyapatite.
4. X-ray diffraction showed foils with hydroxyapatite peaks along with hydroxyapatite peaks along with the substrate titanium peaks.
5. Scanning electron microscopic study revealed micropores and scaly appearance of the uncoated foil, and uniform coating of hydroxyapatite.
6. The surface roughness and pores has given provision for the attachment of the hydroxyapatite crystals.
7. Osteoblasts adhesion test was done and scanning electron microscopic study was conducted to view the adhered cells.
8. The osteoblast cells have adhered to the substrate of both the coated and uncoated foils.
9. At higher magnification spreading of an individual cell was seen.
The results reveal that this study could give rise to a new generation of osseoconductive membranes which can be used in various clinical conditions.
Englishhttp://ijcrr.com/abstract.php?article_id=2310http://ijcrr.com/article_html.php?did=23101. Hasegawa, Jan-Thorsten Schantz, Dietmar et al, Evaluation of a tissue-engineered membrane-cell construct for guided bone regeneration. INT J ORAL MAXILLOFAC IMPLANTS 2002; 17 : 161 – 174.
2. Hurzeler, Francisco H. Nociti, et al, Absorbable versus Nonabsorbable membranes and bone autografts in the treatment of ligature-induced peri-implantitis defects in dogs: A histometric investigation. INT J ORAL MAXILLOFAC IMPLANTS 2001; 16: 646-652.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14TechnologyGROWTH AND CHARACTERIZATION OF PICRIC ACID MIXED ZTS SINGLE CRYSTALS
English122132N.Balasundari English P.SelvarajanEnglish S.Lincy Mary Ponmani D.JencylinEnglishThe growth of picric acid-doped Zinc tris (thiourea) sulphate ( ZTS) single crystals were grown from aqueous solution by slow evaporation technique. 1.5 mol% of picric acid was added in saturated solution of ZTS. When picric acid was added as dopant, morphological alterations were noticed in the grown crystals. Cell parameters of the grown crystals were obtained from the XRD analysis and the presence of functional groups was identified by FTIR study. Its optical properties were examined by UV-vis analysis. The microhardness values were measured for the grown crystals and discussed.
EnglishZTS; doped ; Solubility; crystal growth; FTIR; microhardness;SHG.1. INTRODUCTION
Zinc (tris) thiourea sulphate (ZTS) is a semiorganic NLO material for second harmonic generation from metal complexes of thiourea. High-damage threshold and wide transparency make it a better alternative for KDP crystals in frequency-doubling and laser fusion experiments [1,2]. SHG efficiency of ZTS crystal is 1.2 times more than that of KDP [3,4]. ZTS crystal belongs to the orthorhombic system with the space group Pca21. The growth of ZTS crystals from aqueous solution and its characterization have been reported in a number of recent publications [5-9]. Picric acid is one of the organic compounds having tendency to form the stable picrate compounds with various organic molecules. It has been reported that doping NLO crystals with organic and inorganic additives can alter various physical properties and doped–NLO crystals may find wide applications in optoelectronic devices compared to pure NLO crystals [10,11]. Keeping this in mind, pure and picric aciddoped Zinc (Tris) Thiourea sulphate (ZTS) crystals have been crystallized and studied in the present work. The results of the growth, solubility, XRD studies, FTIR studies, UV-Vis analysis, and microhardness studies of the grown crystals are reported in this paper. 2.Experimental methods and instrumentation 2.1.Synthesis and Growth Pure crystal of Zinc (tris) Thiourea sulphate (ZTS) was synthesized by dissolving high purity AR grade Zinc Sulphate heptahydrate and thiourea in molar ratio 1:3 in double distilled water. The solution was stirred by a magnetic stirrer and white crystalline ZTS salt was obtained on heating at 45 oC for a long time. The salt was purified by repeated recrystallization. To obtain picric acid-doped ZTS salt, 1.5 mol% of picric acid was added to solution of ZTS and the solution was heated at 60 oC . Single crystals of ZTS and picric aciddoped ZTS were grown from saturated solutions of synthesized salts by the solution growth employing slow evaporation technique at room temperature (31 oC). The good quality transparent and colorless pure ZTS crystal was harvested in 20 -25 days. Transparent and pale yellow colored picric acid-doped ZTS crystals were harvested within 20 days. The photograph of grown crystals is shown fig. 1.
2 Instrumentation Single crystal X-ray diffraction studies were carried out for the grown crystals of this work using ENRAF NONIUS CAD-4 X-ray diffractometer with MoKα (λ=0.71069 Å) radiation to evaluate the structural properties. The transmission properties of the crystals were examined using Lambda 35 model Perkin Elmer double beam UV-vis-NIR spectrometer in the range from 190 nm to 1100 nm. Optically polished single crystal of thickness 2 mm was used for this study. The Fourier transform infrared spectrum (FTIR) of the sample was recorded in the region 400-4000 cm-1 with Perkin Elmer Fourier transform infrared spectrometer (Model : Spectrum RXI) using KBr pellet. Microhardness measurement was carried out using Vickers microhardness tester fitted with a diamond indentor. The Second Harmonic Generation (SHG) conversion efficiency was tested using a set-up of Kurtz and Perry [12] and it was carried out using Qswitched mode locked Nd:YAG laser with first harmonic output at 1064 nm. The grown sample was powdered with uniform particle size of about 600 ?m using a ball mill and the powdered sample was packed densely between two transparent glass slides. The fundamental laser beam of 1064 nm wavelength, 8 ns pulse with 10 Hz pulse rate was made to fall normally on the sample cell. The emitted green light from the sample was detected by a photomultiplier tube (PMT) and displayed on a Cathode Ray Oscilloscope (CRO). KDP crystal was powdered into identical size as that of the sample of this work and it was used as reference material in the SHG measurement. 3.Results and discussion 3.1.Solubility studies Solubility study for the synthesized salts was carried out using a hot-plate magnetic stirrer and a digital thermometer. The procedure for finding solubility is reported elsewhere [13]. The variation of solubility with temperature for the samples is shown in figure 2. It is observed from the results that the solubility increases with temperature for both the samples and it is found to be more for picric acid doped ZTS sample. It is clear that for the picric acid -doped sample, the solvent is able to accommodate a marginally increased amount of solute for the saturation at the same temperature. Since solubility increases with temperature, the samples of this work have positive temperature coefficient of solubility. The increase in solubility for the picric acid-doped sample may be responsible for the change in the growth rate and morphological change observed in the present work.
3.2. Single crystal XRD studies
The grown crystals were subjected to single crystal XRD analysis and single crystalline data were obtained. From the data, it is observed that pure and picric acid-doped ZTS crystals crystallize in orthorhombic system and the unit cell parameters are listed in table 1. The obtained values of lattice parameters for the pure ZTS crystal are found to be in good agreement with the reported literature [5]. The changes in the lattice parameters are due to incorporation of picric acid in the lattice of ZTS crystal.
3.2 Powder XRD Analysis
The powder X-ray diffraction (XRD) pattern picric acid-doped ZTS crystal are shown in the figure 3. The well-defined peaks at specific 2θ values show high crystallinity of the grown crystals. All the reflections of powder XRD patterns of this work were indexed using the TREOR software package following the procedure of Lipson and Steeple [14]. The values of hkl, relative intensity and 2 theta values for the reflection peaks of the powder XRD pattern are given table 2. Using the values in the table 2 and the UNITCELL software package, the cell parameters were found and it is observed that the values of unit cell parameters obtained from powder XRD method are in comparable with those obtained from single crystal XRD method.
3.3 FTIR Spectral studies The FTIR analysis was carried out by Perkin Elmer FTIR spectrometer by KBr pellet technique in the range 500-4000 cm-1 . The FTIR spectra of pure and picric acid mixed ZTS samples are shown in figure 4. In ZTS complex, there are two possibilities by which the coordination with metal can occur. It may be either through nitrogen or through sulfur. From spectra, the N-H, absorption bands in the high frequency region in thiourea were not shifted to lower frequencies on formation of metal thiourea complex, thus coordination of thiourea occurs through sulfur in ZTS [11]. The NH, C=S and N-C-N stretching vibrations were also seen. The comparison of the samples shows slight shift in characteristic vibrational frequencies of 1.5 mol% picric acid doped ZTS with respect to pure ZTS. This confirms the addition of picric acid in grown crystal. The spectral assignments for the picric acid mixed ZTS sample are provided in the table 3.
3.4.UV-Visible spectral study The study of optical transmission range of grown crystals is important because the crystals are mainly used for optical applications. The UV–visible study of grown crystal was carried out by Varian-Cary 5E UV-vis Spectrometer in a range 200 nm to 1100 nm. The absorption spectrum of 1.5 mol% picric acid–doped ZTS is shown in fig.6 and the inset figure shows UV spectrum of pure ZTS for comparison. The absorption spectrum reveals that the crystal has lower cutoff wavelength at around 290 nm. The absorption near UV region is associated electron with transition within thiourea units of ZTS. Doping of 1.5 mol % of picric acid in ZTS does not destroy the transparency of the crystal. From spectra it is observed that the lower cutoff wavelength is almost the same for picric acid doped ZTS and pure ZTS crystals. The wide range of transparency in UV-visible and IR region enables good transmission of the second harmonic frequencies of Nd:YAG laser. This is an added advantage of the grown crystals of this work in the field of optoelectronic applications.
3.5.Measurment of microhardness
Microhardness of a crystal is its capacity to resist indentation. Physically hardness is the resistance offered by a material to the localized deformations caused by scratching or by indentations[15].The selected smooth surfaces of the grown crystals were used for microhardness measurement at room temperature using a Vickers microhardness tester fitted with a diamond indenter. The hardness of the crystal is calculated using the relation The hardness of the crystal is calculated using the relation Hv = 1.8544 P / d2 kg / mm 2 where P is the applied load in kg and d is the length of indentation impression in millimeter and 1.8544 is a constant of a geometrical factor for the diamond pyramid [16]. A plot drawn between the hardness value and corresponding loads is shown in figure 6. It is noticed that Vickers hardness number (Hv) increases with the applied load satisfying the indentation size effect. When a ZTS crystal is doped with picric acid, the hardness decreases and this decrease in the hardness value of doped sample can be attributed to the incorporation of picric acid in the lattice.
4. Conclusion
Single crystals of pure and picric aciddoped Zinc (Tris) Thiourea sulphate(ZTS) were synthesized and solubility studies were carried out for the prepared samples in de-ionized water in the temperature ranging from 30 to 50 oC. In accordance with the solubility data, saturated solutions were prepared for growing pure and picric acid doped ZTS crystals by slow evaporationtechnique. Morphological alterations have been observed when ZTS crystals are doped with picric acid. The powder and single crystal XRD studies confirms the orthorhombic structure of the grown crystal..The unit cell parameters of pure ZTS crystals are in agreement with the literature values. The FTIR spectrum confirms the presence of all the functional groups in the grown crystals. UV-visible study reveals the picric acid doped ZTS crystal has lower cutoff wavelength at around 290 nm. Mechanical property of the grown crystals has been studied by microhardness test and noticed that there is a decrease of microhardness number for 1.5 mol% of picric acid-mixed ZTS crystals. Acknowledgement The supports extended in the research by Sophisticated Analytical Instrumentation Facility (SAIF), IIT, Chennai, RRL, Trivandrum and M.K.University, Madurai are gratefully acknowledged. We thank authorities of Aditanar College of Arts and Science, Tiruchendur and Infant Jesus College of Engineering, Keela Vallanadu, Tuticorin for the encouragement given to us to carry out the research work.
Englishhttp://ijcrr.com/abstract.php?article_id=2311http://ijcrr.com/article_html.php?did=23111. P. Kerkoc, V. Venkataraman, S. Lochran, R.T. Bailey, F.R.Cruickshank, D. Pugh, J.N. Sherwood, J. Appl. Phys. 80 (1996) 6666.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesOCCUPATIONAL AWARENESS OF RURAL WOMEN WORKERS IN HEN POULTRY FARM, COIMBATORE DISTRICT, SOUTH INDIA
English133140Sharmila Banu AEnglish Gunasekaran CEnglishSelvaKumar VEnglish Mohana PEnglishKandappan English KEnglishElanchezhian MEnglishFrequently the rural area women?s are first-rate the preference to poultry works. Especially women?s are the
majority to work on this field. Rural areas are mainly depends on the women incomes. The present research focus to
scrutiny the occupational awareness in hen poultry farm, in specific reference to faecal cleaners of women workers.
The foremost objective of the research is to bestow the awareness about work place and mould with them health and
safety decisive factor. The calculation is done by percentage (%) intensity and chi-square test. The recommendations
in addition discuss among women workers.
Englishoccupational awareness, rural, hen faecal cleaning women, livelihood, percentage and chi-square test.INTRODUCTION
Even though they are home makers, but they are forced to work for their family. This is help to their children future and they have a thought about empowerment development. The women are the backbone of agricultural workforce but worldwide her hard work has mostly been un paid. She does the most tedious and back-breaking tasks in agriculture, animal husbandry and homes (DARE/ICAR ANNUAL REPORT 2003–2004). The choice of occupation is traditionally (Penny Kane) or time being taken to commencing of rural women?s and they have the poor dexterity of occupation effects. Thus, the rural women?s are randomly selects the hen poultry farm. In particularly, we have one question: Why, the research is to selects the poultry farm and especially in women workers. Because, POULTRY is by far the largest livestock group and is estimated to be about 14 000 million, consisting mainly of chickens, ducks and turkeys (FAO 1999).One third of the world is moving only the food of chicken. Women are the back fillet of the whole lot in family. She have to work for all the relationships like Mother, Daughter, Sister, Grand mother, worker in occupation etc. She didn?t feel difficulty with anything which comes from family or occupation. The bacterial count in poultry housing systems is high in comparison to those of pig and cattle. Little is known about the bacteria present in the poultry environment such as in poultry litter and air of poultry house (Saleh et al., 2003, Nasrin etal). Micro organisms like Bacteria, (USA Wrestling 2007-08) fungi-Small organisms that are found everywhere in the environment and on skin. Whereas their population is more high in pollutants and excrete area like animal faeces. These organisms move under the skin and start the problem to the humans. In poultry farm may cause the problems of infection disease respiratory problems and so on, these overwhelming effects to the worker when they have not any idea about basic occupational skill (safety manners). The possible negative effect of noise on reproduction is biologically plausible, as well as amenable to prevention (Irene Figà-Talamanca). So the research have in mind of these points of basic health problems and its root will be spread when the daily habits are not in following the proper comportment such as clean dress, wear the gloves and mask, and footwear. Since, these are the fundamental to hygienic working people. The present research asked the question about clean dress, wearing the gloves, mask, footwear?s and hand washing habits. These all done by direct interview method. This analysis is calculated by percentage (%) variation. Based on the results they urge.
METHODOLOGY
The research work was conducted on the rural women worker in hen poultry farm in Coimbatore District. After approval from Institutional Ethics Committee,(Kartik et al 2005) the research was on track with conquering an informed permission from the partaker of poultry farm. Totally 130 samples were selected for this research which included age group of Up to 25 with 29 samples (group I), 26-30 with 74 samples (group II), 31- 35with 19 sample (group III), 36 an above with 8 sample (group IV) and the study was conducted during the epoch 3rd -24th May 2012.The study was vigilance by direct interview method, we calculated the percentage of women?s worker awareness through the questionnaire include the 5 basic question, this also designed by the points of daily activity such as clean dress code, wearing gloves and mask , footwear then washing habit of hands, and the answers were corrected with options of 1.Yes, 2.No then this research was successfully completed by the percentage and chisquare test calculations. The Direct interview Questions are following A. Are they workers maintaining the dress code (a clean pair of working dress) to work place? B. Are they having the habit to wear footwear in work place? C. Are they wearing the disposable gloves? D. Are they use the mask at work place? E. Are they washing hands before drinking water? The standard optionally answers were assessed directly by the research team. Based on the options the findings were examined.
DISCUSSION
The present research was focused to identified the occupational awareness in rural area especially women workers in hen poultry farm. In the attendance the research encounter only the fecal cleaners. Why? The study concentrated to faecal cleaners because they were more infected through the excrete materials in poultry farm ,According to Mojtaba Yegani (2010)Zoonotic diseases are transmitted from animals to humans and include bacterial, viral, fungal, and parasitic diseases. Salmonellosis, campylobacteriosis, chlamydiosis, tuberculosis, Newcastle Disease, and avian influenza are amongst the most common zoonotic diseases transmitted from poultry to humans. Poultry workers are at a greater risk of being affected by these diseases. This review also support to our research. Based on the results the age group of up to 25 was more aware worker than the rest of the age group. Why? The research team asked the question about clean dress means some worker have the habit to wear pair of dress for work time, some of the worker have the habit to wear clean dress in daily basis, but few of the workers used the dress for two day or three days. The table A. question only 12 samples answered and the percentage was 9.2%, High percentage was 90.8% found from 118 samples. Similar to the second question about the habit of foot wear, why? We had chosen this, because any time every minute whenever we want anything we used only our travel pack of legs, sometime we may go hospital for children or elders and reach our home without footwear means it can spread the growth of microbial diversity through work place. The table B shows in the percentage of 21.5 from 28 samples and high percentage from 102 samples. The third question asked about gloves, why? The research team set this kind because hands are the direct conduct to get the infection; through the use of hand they cleaned the fecal ooze material. This material has the quality to induce in the skin and cause diseases and hands are the machine for everything like eating, cooking, washing etc. without proper practice in the work place we may get the foundation of ill. So cleaning is the main factor of the above. The table C shows the percentage variation like start from 24 samples of 18.5 and the 81.5 percentage from 106 samples. It.s basic. It.s simple. The number one way to improve health in rural areas is not to provide clean water; that comes in third. It is much easier than that: teach people how to wash their hands (Mihelcic, 2003). Third world communities suffer greatly from preventable water borne diseases and from diseases spread through poor hygiene practices. By improving sanitation and environmental conditions in the communities and implementing an educational program of health and hygiene education appropriate for the community, the numbers of preventable deaths and illnesses have been shown to decrease (Panchita Paulete,2003). According to WHO guidelines for indoor air quality: dampness and mould, Exposure to microbial contaminants is clinically associated with respiratory symptoms, allergies, asthma and immunological reactions. The microbial indoor air pollutants of relevance to health are widely heterogeneous, ranging from pollen and spores of plants coming mainly from outdoors, to bacteria, fungi, algae and some protozoa emitted outdoors or indoors. They also include a wide variety of microbes and allergens that spread from person to person. There is strong evidence regarding the hazards posed by several biological agents that pollute indoor air; however, the WHO working group convened in October 2006 concluded that the individual species of microbes and other biological agents that are responsible for health effects cannot be identified. This is due to the fact that people are often exposed to multiple agents simultaneously, to complexities in accurately estimating exposure and to the large numbers of symptoms and health outcomes due to exposure. The exceptions include some common allergies, which can be attributed to specific agents, such as house-dust mites and pets. So, it?s our duty to be safe in work place and in environment, based on this statement the research team frames the question about wearing of mask at work place, this will help to control the respiratory problems. The table D showed the percentage of 58 samples 44.6 and 72 samples 55.4.Improved hygiene (hand washing) and sanitation (latrines) have more impact than drinking water quality on health outcomes, specifically reductions in diarrhea, parasitic infections, morbidity and mortality, and increases in child growth,.(Water, sanitation and hygiene, 2002).the research team included the question about washing hands before drinking water or any intervals based on the answered from the samples the finding were done. The table E shows the 35.4 percentage from 46 samples and 64.6 from 84 samples respectively. The chi-square test valuable were calculated by the factors of age and awareness, income and awareness, experience and awareness. The Pearson chisquare value was 14.75, Degrees of freedom was 3 and p value was .002(less than .05) these were the values of age and awareness. The income and awareness was Pearson chisquare value 20.155, degrees of freedom 3 and p value was .000(less than 0.05).The experience and awareness were Pearson value 0.817, the degrees of freedom was 2 and p value was 0.665.From the chi-square results the research find out that experience is not mould the person directly but some cases it can. Age is the factor this is have the ability to know that what we will do or what we didn?t. Because the research were not get the significance in experience where as the significance appear in age and income compared with awareness. Hygienic and health conscious to support our immune actions. The research questions such as wearing footwear, to avoid the microbial infection in legs and foot and the questions of gloves and mask were support to the workers from respiratory problems like sneezing, cough and cold. Washing hands and wearing clean dress are important to live good life. These are the basic questions arise from the research team and observed by direct interview method. This is the first stage in a process that helps empower members of a community. To assess their knowledge base, investigate the local environment, visualize a future scenario, analyze constraints on change, plan for change, and implement it,. (PHAST, 1998).Based on this review support, our research was analyzing the occupational environment on rural women workers in first stratum.
CONCLUSION
During the month of period the research was conducted on hen poultry with randomly selected 130 samples in various poultry farm. Whenever, we interview the women workers by directly that time on spot awareness created to them its shows visibly. After two or four hours the awareness was disappear. This is because of the care less and not giving the preference to health and long life. But awareness not only the source to protect them, apart from this self appraisal with awareness of each individual concentration is more important than the voice arise from the researcher. As a result we entitled the aphorism to their hearts that is REALIZE your work and know the REALITY of your occupation, THIS WILL REDUCE THE OCCUPATIONAL PROBLEMS of health.
ACKNOWLEDGEMENT
I thank to God to have this opportunity to do something good to society especially women workers, Our sincere thanks to Dr.Sasikala, head and professor, Department of Zoology, Bharathiar University, with her support and encourage completing this work in smooth way. Gratitude to Institutional Human Ethical Committee of Bharathiar University for permit the research work. Our heart full thanks to poultry farm women workers for their patience and managing director, supervisor and last but not least Self Help Group members for their support in this field. Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2312http://ijcrr.com/article_html.php?did=23121.FAO 1999. Statistical database, www.fao.org. Food and Agriculture Organisation of the United Nations. Rome,Italy.Houghton, MI.
2.Irene Figà-Talamanca Occupational risk factors and reproductive health of women Department of Hygiene and Industrial Health, University of Rome „La Sapienza?, Piazzale Aldo Moro 5, 00185 Roma, Italy. e-mail: irene.figatalamanca@uniroma1.it
3.Jim Porter, USA Wrestling MRSA and other Infection Facts Making Wrestling Safer Guide to Recognition of Skin Infections james.porter@selectmedicalcorp.com
4.Kartik R Shah and Rajnarayan R Tiwari 2005, OCCUPATIONAL SKIN PROBLEMS IN CONSTRUCTION WORKERS Indian J Dermatol. 2010 Oct-Dec; 55(4): 348– 351.doi: 10.4103/0019-5154.74537
5.M.S. Nasrin, M.J. Islam, K.H.M.N.H. Nazir, K.A. Choudhury and M.T. Rahman1 Identification of bacteria and determination of their load in adult layer and its environment Dept. of Microbiology and Hygiene, Bangladesh Agricultural University, Mymensingh-2202, Bangladesh J. Bangladesh Soc. Agric. Sci. Technol., 4(1 and 2): 69-72, 2007 ISSN 1811-6221
6.Mihelcic, Jim, Ph.D. Lecture. (13 March 2003).Michigan Technological University. Houghton, MI
7.Mojtaba Yegani, 08 Jul 2010 Prevention / Control Health hazards: Safety always comes first, University of Alberta, Canada
8.Panchita Paulete (May 2003), Practice and importance of combining health/hygiene and environmental education with water sanitation for sustainable health benefits in rural areas School of Forest Resources and Environmental Science Michigan Technological University, Houghton, MI
9.Penny Kane, Senior Associate, Editor WOMEN and OCCUPATIONAL HEALTH, Issues and policy paper prepared for the GLOBAL COMMISSION ON WOMEN'S HEALTH The Office for Gender and Health, The University of Melbourne, Australia
10.PHAST Initiatives in East Africa. (1998). The World Bank. Presented at the Community Water Supply and Sanitation Conference,Washington, DC.http://www.wsp.org/english/focus/conference/ hyg_phast.pdf SOPAC WRU Strategies
11.Saleh, M., Seedorf, J. and Hartung, J. 2003. Total count of bacteria in the air of three different laying hen housing systems. Dtsch. Tierarztl. Wochenschr., 110(9): 94-97.
12.Water, sanitation and hygiene: at a glance..(March 2002). World Bank Group. http://wbln0018.worldbank.org/HDNet/hdd 60/9d1422d8016e85d885256b90005e1f76/$FILE /5WatSan%20AAG.pdf
13.WHO guidelines for indoor air quality :dampness and mould This document presents World Health Organization (WHO) guidelines for the protection of public health from health risks due to dampness, associated microbial growth and contamination of indoor spaces oct 2006
14.WOMEN IN AGRICULTURE, (DARE/ICAR ANNUAL REPORT 2003–2004)
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareFUSION FIRST AND SECOND THORACIC RIB NEAR STERNAL END
English141144Londhe ShashikalaEnglish Kori RohiniEnglish Panjakash SamreenEnglishDuring routine work in the osteology section in the department of anatomy, Al- Ameen medical college Bijapur, we got an unusual specimen showing fusion of first and second rib near its sternal end on left side. Fusion was along the outer margin of first rib and inner margin of second rib, 2.2 cm and 2.8cm behind the costo-chondral junction, along the inner and outer margin of first and second ribs respectively.
Fused segment was 8.5cm in front of vertebral end along outer margin of first rib. 8.2cm in front of vertebral end along inner margin of second rib. Transverse Length of fused part between first and second
rib was 3.3cm and length of intercostal space in front and behind the fusion was 0.6cm and 1.1cm respectively. Both the ends of both ribs were free so intercostal space at the costochondral junction and at
the vertebral end was open. Because of such type of rib variations there are chances of compression of
neurovascular bundle along the intercostal space. These fused rib affect chest wall expansion, may result
in respiratory complications. Knowledge and awareness of such anatomical variation of rib is important
for physicians, radiologist and surgeons.
EnglishFusion, first and second thoracic rib.INTRODUCTION
There are two types of variations related to ribs, numerical and structural. In numerical type, number of ribs either more or less. Cervical or lumbar rib may be present as extra ribs, one or more ribs may be absent as in less ribs. In structural type, there is bifid rib, short rib and fused rib.1 Each rib originates from the caudal half of one sclerotome and the cranial half of the next subjacent sclerotome. Head develops from sometocele cells from one somite which migrate with the caudal half of sclerotome, proximal portion of the rib develops from both caudal and cranial sclerotomal halves, and there is no mixing of cells from these origins. Distal portion of rib developed from caudal and cranial halves of sclerotome, these cells mix as the rib extends into the ventral bodywall and segmentation diminishes .2 Any change or variation at this step of development may be the reason for fusion of ribs . Irregular segmentation of primitive vertebral arches lead to fusion anomalies, recent experimental studies in mice have implicated splotch gene mutation for various structural abnormalities in the ribs3 . Bicipital rib is an unusual anatomical pecularity which results due to the fusion of shaft of two distinct ribs into a common body and is seen exclusively in relation to the first rib, either due to fusion of cervical rib with first rib or more commonly due to fusion of the first rib with second rib4, 5. Its incidence has been reported o.3% in a study based on chest radiograph. 5 The rib anomalies whether pathological or normal varients such as cervical rib, pelvic rib, bifid rib and bicipital rib etc. often indicate an underlying systemic disorder.6 CASE REPORT We noticed a left sided bone specimen showing fusion of first and second ribs, during routine work in the osteology section of department of anatomy, Al- Ameen medical college Bijapur .Specimen was examined in detail for its anatomical features, measurements were done and photograph taken from both aspects of rib as in figure 1. Complete length of first rib along the outer margin was 13cm, complete length of second rib, along the inner margin was 14.3cm. Fusion of two ribs was near the sternal end, fused segment was 2.2cm and 2.8cm behind the costochondral junction of sternal end, along the inner and outer margin of first and second rib respectively. Fused segment was 8.5cm infront of vertebral end along outer margin of first rib and 8.2cm infront of vertibral end along inner margin of second rib .Transverse length of fused segment was 3.3cm. The remaining part of intercostal space, before and after the fused segment of rib was open. The first rib had normal anatomical features, that is head, neck and tubercle of vertebral end was normal and at the sternal end it had depression at the tip for costal cartilage. Along the outer margin of first and second ribs there were prominent impressions for serratus anterior muscle towards the vertebral end. The second rib had rudimentary vertebral end, head, neck and tubercle was not prominent and sternal end was cut. DISCUSSION The first rib anomalies include the floating rib, central defects bridged by ligamentous band , rudimentary structure terminating in a synostosis or pseudoarthrsis with second rib, bifurcated rib etc.7Vinita Gupta et al reported partial fusion of rib in anterior and posterior portion and completely blended with each other in middle ,the first intercostal space was obliterated8 .Rib fusion causes scoliosis and restriction of chest wall expansion, which may require surgical correction. Fused ribs also encountered in Gorlin syndrome.6 Rani Anita et al reported synostosis between the anterior ends and shafts of 1st and 2nd ribs of right side. There was single articular oval facet on head with scalene tubercle at the inner margin of shaft of upper segment1 . David Levi reported a case of 8 year boy with lump at inner end of first right rib. There was history of injury, the skiagram showed fusion of first and second rib in the middle third of their curve, he stated that condition appeared to be congenital. He also stated that when fusion is at the sternal end is due to ossification at the costochondral joint, it spread towards the sternal end of ribs, when fusion occurs at the vertebral end, it is due to lesion of intrathoracic content, especially long standing collapse of lung. Fusion of middle third is a rare condition for that no instance is recorded in anatomy textbooks9 . Any scoliosis associated with fused rib may result in three dimensional thoracic deformity with adverse effect on thoracic growth and function with development of thoracic insufficiency syndrome, which is defined as the inability of the thorax to support normal respiration or lung growth. He also stated that children suffering from thoracic insufficiency syndrome referred to them were on oxygen or ventilator support, in all of these patient the common denominator was a small, stiff distorted thorax that could not provide volume for growth of the underlying lungs and functioned poorly, with minimal secondary breathing mechanism because of fused, malformed or absent rib.10 Baumgarlner F et al reported two cases of fused first and second ribs associated with thoracic outlet syndrome in a radiological study.11 Seema Deepak reported an unusual case of a bicipital rib in which first and second rib of right side were fused obliterating the intervening intercostals space, she concluded that such a specimen cannot present with thoracic outlet syndrome but can also be an indicator of an underlying systemic disorder.12
Englishhttp://ijcrr.com/abstract.php?article_id=2313http://ijcrr.com/article_html.php?did=23131. Rani Anita, Rani Archana, Chopra Jyoti, Manik Punita.Synostosis of First and Second Rib- Case Report.J. Anat. Soc. India. 2009;58(2):189-91.
2. Standrings,Johnson D, Ellis HAND Collin P.Grays Anatomy, 39th edition, Churchill Livingstone, London,2005.pp 796.
3. Evans DJR. Contribution of somatic cells to the avian ribs. Developmental Biology.2003;256:114- 26.
4. Turner WM. Cervical ribs and the so called Bicipital ribs in man in relation to corresponding structures in the Cetacea. Journal of anatomy and physiology. 1883;17(30)384-400.
5. Terry RJ, Trotter M. The Ribs, In: Schaeffer JP (eds) Human Anatomy, A Complete Systemic treatise 11th edition. The Blakiston Company, New York, Toronto, 1953, pp.192-98.
6. Glass RB, Norlon KI, Milre SA, Kaug E. Pediatric ribs:a spectrum of abnormalities. Radiographics.2002; 22: 87-104.
7. White JC, Poppel MH, Adams R. Congenital malformation of the first thoracic rib: A case of brachial neuralgia which simulates the cervical rib syndrome. Surg Gynecol Obstel.1945; 81:643-59.
8. Vanita Gupta, Rajesh Kumar Suri, Gayatri Rath, Hitendra Loh. Synostosis of first and second thoracic ribs: Anatomical and radiological assessment. International journal of anatomical variations. 2009;2:131-33.
9. David Levi. Fusion of middle third of the first and second ribs. Proc R Soc Med. 1932; 25(8)1233.
10. Robert M. Cambell JR, Melvin D. Smith,Thomas C.Mayes,John A. Mangos,Donna B. Willey-Courand, Nusret Kose, Ricardo F. Pinero,Marden E.Alder, DDS,Hoa L. Duong, Jennifer L.Surber, BS.The characteristics of thoracic insufficiency syndrome associated with fused ribs and congenital scoliosis.The journal of bone and joint surgery, Jbis org.203;85(3):399- 408.
11. Baumgarlner F, Nelson RJ, Robertson JM.The rudimentary first rib :Acause of thoracic outlet syndrome with arterial compromise. Arch Surg. 1989; 124:1090-92.
12. Seema Deepak, Dikshayani KR. An unusual case of a bicipital rib- a case report.Anatomica, Karnataka.2011; 5(1)50-52.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14General SciencesCONSTRUCTION OF THE MATHEMATICAL MODEL FOR EVALUATION AND FORECAST OF INTERURBAN
PASSENGERS TRAVEL
English145152Eng. Shkelqim GJEVORIEnglishFor a more harmonious development between economy and transport and for a close relation supply and
demand for travel, proper scientific studies should be conducted to determine operational instruments for
resolving the contradictions between this report and especially those technical instruments should rovide recommendations on permanent and long-term planning that preceded the decision and policy-makers. One of these instruments that are in use with much interest and after 50 years are mathematical models in transportation. They are based on the assumption that a linear equation could represent the relationship between a dependent variable “Y “ that represents a pointer or economic phenomenon and one or more independent variables “ X , X ....X p 1 2 “ and that each of them taken separately can satisfy the assumption of linearity that is expressed in mathematical form as follows, y F a a X a X a X e n n ? ? ? ?. ? ?...... ? 0 1 1 2 2 (1.0) It is important that we have implemented practical use of information which is part of a database system for collection and data management of travel and road traffic in our country. In particular and very carefully, we have also defined specific requirements for input data analysis and have made their selection for the purpose of constructing the model.
EnglishO/D matrix, demand modelling, supply, variables, balance.INTRODUCTION
Theoretical treatment and research on our part has been extended by examining all transport?s components, taking into account the current information system that forms the basis of existing data, and providing other forms of data impossible to ensure up to now but that are indispensable. On the other hand refereed to the specific conditions of Albania's research and collection of appropriate data is not easy mainly because of the fragmented characteristics of existing statistical data and lack of time series. Basic variables which we are supported affect the quantity and quality of travels and we have grouped into three categories as follows, demographic, economic, territorial factors.
MATERIALS AND METHODS Based on three categories of the above variables from an analysis of their grouping, have deemed to representation and application of the travel modeling, the following variables as, resident population in cities, the number of passenger vehicles, and the active forces able to work, economically active enterprises, number of employees. With travel ensuring O/D (origin-destination) between the cities of the country and variables expressed above we have ensured the necessary database for the travel modeling through the application of selected mathematical model. To derive the travel legality in the function of variables "Xi" selected according to the recommendations and analysis made by us, we were focused on some of the country's main towns which make up about 96 % of the population and economic activities, of variables "Xi" taken into consideration. Starting from the given travel data O/D and for the 5 selected variables we have draw 5 tables. Starting from the given travel data O/D and for the 5 selected variables we have draw 5 tables, travels O/D depending on 1.the population variable (Xp), 2.number of economically active enterprises (Xnd), 3.vehicle of transportation for passengers (Xmj), 4.active work force (Xap) and 5.number of employees.(Xnp) as an example only one of them (table 1) for, a) selection of independent variables classifying by weight occupied by each of them, and to make a second selection of the 5 variables in the three we have applied the mathematical model of the shape force, 2 1 c Y ? c X (1.1) and for, b) modeling and forecast of interurban trips we shall apply the mathematical model of linear form, Y a a X a X a X e ? 0 ? 1 1 ? 2 2 ? ?... n n ? (1.2) Mathematical model of the selected form Y = c1X c2 is based on comparing of the process conduct and this of the model, especially when the dependence between the "Y" and "Xi" is not linear and more so when there is no identity of conduct. The objective of the method is to modify the model parameters in order to achieve a better fit by assuring a very fast convergence and adjacent to the redevelopment status and optimal. The result of the model describes a relationship between average variables of the two variables. The results given by the model are tabular and in graphical form, showing except of the parameters values (coefficients before variables "Xi"), and some important indicators that show the reliability of the model and extent of links between the two sizes "X and Y" such as, R, R 2 (or R 2 correlation coefficients, an indication that express the value of the model), adjusted R 2 (or Adjuster R) is useful in comparing this model with other models, F (value of the coefficient of reliability, Pvalue (probability value of reliability of coefficients before variables "Xi", etc. Based on the selected mathematical model we have applied software of this model (1.1) for nine main cities like Tirana, Durres, Vlora, Elbasan, Fieri, Korca, Berat, Shkodra, Lezha, representing cities centers of the county towns that constitute the major economic, social and cultural potentials, taking as the origin one city and destination eight other cities and so changing the positions of origin of travels to eight cities in an order we have received the following results which express the legality of dependency between variables "Xi" and function "Y". In here we are providing only one of the results of application of software. Journeys in the function of the population origination from Elbasan. (table 1/1) where; TT(pop)-is variable “Xp” that gives the number of population for cities Berat, Durres, Fier, Korca, Lezha, Shkodra, Tirana and Vlora (which are marked with the indices according to the vehicles number plates of the respective cities). UP(O/D)-are current journeys originating travel between the city of Elbasan and 8 other cities of destination (which in fact is the value of the function of “Y” mathematical model that we implemented) YP(legality), is the result of the travel legality "Y" according to the application of mathematical model. From the results of model we are taking, Fopt = 3.762306016217445E-002 Copt(1)=2.658623009781310E007 Copt(2)=1.790759920467837 where; Fopt-function value (Y legality) Copt(1), Copt(2)- values of model parameters. In that way the function has the form, 1.790 Y ? 0.0000002658X p (1.3) In the same way we extracted the results and for eight other cities taken by order as the origin of travels. In this way the characteristics of transport in urban areas reflect in a broad way the social, economic, physical conditions at a given point in a given time. In Table 2 are given the variables and the correlation?s coefficient. Thus from the 200 directions of travel between 8 cities in the study, about 80-85% of them express a reliable correlation between the curve of O/D and that of legality of the O/D found by modeling the remaining 15-20% displays a correlation not very satisfactory. To select which of the variables “Xi” influence more on trips O/D, it was applied their simulation “Xp, Xmj, Xnd, Xfa, Xpu” by increasing them to the extent of 10%. From the application of mathematical model of the type power (equation 1.1) we obtained 200 values of trips O/D between the couple of cities and we get the following results. Specifically for simulation with 10 % of “Xp” we get the results as table 3. As a conclusion based on the above two assessments therefore; 1) to the extent of influence after the simulation with 10% of variables, 2) and coefficient “kO/D/Xi” [travel/unit variable]. Have made their classification, from which we selected three of variables that have significant impact on the generation and attraction of trips which are, population (Xp), vehicle of transportation of passengers (Xmj), and economic enterprises (Xnd) Based on these three selected variables in accord with (equation1.2) "construct" the model of trips generated or attracted in nationally rate, where, e-error term, for X1=Xp,; X2=Xnd and X3=Xmj. Then our function will have the following form, Y = a0 + a1Xp + a2Xnd + a3Xmj + e (1.64) STATISTICAL METHODS. Descriptive statistics, mean and standard deviation were tabulated using SPSS 16.0 software. RESULTS. Based on official data of INSTAT and the values of travel according to the matrix O/D we have benefited Table 4. To obtain the mathematical form of the model for interurban travel in national scale have applied software SPSS, where we have the following results and linear form of mathematical model of interurban travel. According to the results of software and the taken coefficients interurban travel modeling has the form of general equation as follows, Y=1003.578–0.029Xp+1.68Xnd+0.632Xmj (1.7) Based on the above model we calculated the values of the function “Y“ for 22 centering (cities) and the results are given in table 5. In this case, model rejected by the negative value of coefficient “a1 = -0.029 ” variable that is before "Xp" which is impossible, because it is in confrontation to the generation theory and trips attraction which estimate that the population is the community of variables that affect positively in increasing the travel. This discrepancy is due to multicorelation among the selected variables by our side with the desire to include as many variables in the modeling of travel. To continue to the realization of the goal of work shall exclude the variable "Xp" from the table 5 above and would appreciate the dependence of travels with two other variables “Xmj,and Xnd”. Besides the elimination of population variables we have increased the number of zone (cities) to have an even greater representation of spatial interaction. To obtain the mathematical form of model for generation interurban trips (table 5) in nationally scale we have applied again software SPSS, where we get the following results and linear form of mathematical model of generating interurban travel. (table 6,7)
DISCUSSION
According to the results of software and coefficients of Table 7, are, a0 = - 329.479; a1= 1.315; a2= 0.535; (1.8) and interurban travel modeling generation has the form of general equation as follows Ygj = - 329.4+1.315*Xnd+ 0.535*Xmj (1.9) Regarding the reliability of function (1.9) we halted in the analysis of some parameters that are evaluated by software through ANOVA (analysis of variation). From the results of tables 6 and 7 we see that the coefficient of determination is R 2=0.97 or 97%, which highlights the extent of variation of the “Y” from two variables “Xmj,andXnd” and from statistical theory is considered as a criterion to judge objectively on the quality of the model because it is the indicator of the degree of approximation of “Y and Xi” likewise, “R” corrected (adjusted R Square) and "R" Multiple, have very high values that attests to a strong correlation between independent variables and dependent variable. The data results of Table ANOVA according the table OUTPUT SUMMARY results that Significance F=1.45705E-20 is less than ? ? 0.05 so we are within a specified condition, while the validity coefficient (importance) to the ? ? 0.05 According the values of table I the book of Statistics) we have that probability of regression coefficient at the variable “Xnd" is 90% as the P-value=0:06, while the probability of the coefficient of regression at the variable "Xmj" is 95% as the Pvalue =0007 so many good values which estimate the model of "construction". Above model parameters carry a significant meaning so the regression parameter a1=1.315 before variable "Xnd" indicates that when the number of enterprises increased by 1% the number of trips generated will increase by 1.31% while for the regression parameter a2=0.535 before variable "Xmj" it shows that when the number of vehicles increased by 1% the number of trips generated will be increased by 0535%. Regarding the parameter a0 = - 329.479 in general have no much sense and requires great care in interpretation.
CONCLUSION
The Design of a model for the evaluation of interurban trips will help decision makers in policy planning, management and investment in the transport field.
ACKNOWLEDGEMENT.
I thank the staff of the Institute of Transport for the support given in data-base as well as colleagues of the Faculty of Mechanical Engineering, Polytechnic University of Tirana. Also thank the authors and co-authors for the literature that was useful in this publication. Authors acknowledge the immense help received from the scholars whose articles are cited and included in the references of this manuscript.
Englishhttp://ijcrr.com/abstract.php?article_id=2314http://ijcrr.com/article_html.php?did=23141. John P. Sammon, Robert J. Caverly 2007. Transportation Systems.
2. Josef Sussman 2005. Introduction to Transportation System.
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5. C.S. Papacostas. P.D. Prevedours 2005. Transportation Engineering and Planning.
6. Prof.Dr. Myslym Osmani. 2005 Statistics.
7. Louis Berger. S.p.a. 2009. Albania National Transport Plan.
8. INSTAT 2009. Indicators by Prefectures.
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10. Michael D. Meyer, Eric J. Miller. 2001 Urban Transportation Planning; a decisionoriented approach.
11. Susan Hanson. The geography of urban transportation. Aug 6. 2004
12. Ryuichi Kitamura, Satoshi Fujii, and Eric I. Pas. Time-use data analysis and modeling: toward the next generation of transportation planning methodologies. 1997
13. David A.Hensher, Kenneth J.Button. 2000. Handbook of Transport Modelling.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareNon-Healing Skin Ulcer in HIV/Tuberculosis Co-Infection: A Case Report.
English153159Monali N. RajurkarEnglish Silpi BasakEnglishIntroduction: Tuberculosis (TB) is very common in India, China and other developing countries. World Health
Organisation (WHO) had estimated 9.2 million new cases of TB, worldwide in 2006 of which 7.7% were positive
for Human Immunodeficiency Virus (HIV). In India, at the end of 2007, there were 2.5 million people living with
HIV and AIDS (PLWHA) whereas incidence of TB was 1.8 million cases per year. Tuberculosis is the most
common HIV related opportunistic infection in India and caring for patients with HIV/TB co-infection is a major
public health challenge. The incidence of tuberculosis is more in people living with HIV infection. So, WHO has
developed the strategy of treating HIV/TB co-infection irrespective of patient?s CD4 count. If any HIV positive
patient is diagnosed to be infected with tuberculosis, the Antitubercular treatment (ATT) is started along with
Antiretroviral therapy (ART).
Here, we report a case of skin ulcer due to Mycobacterium tuberculosis on chest, secondary to pulmonary
tuberculosis in HIV infected person with varied presentation.
Material and Methods: At first pyogenic infection due to Methicillin Resistant Staphylococcus auereus (MRSA)
was diagnosed. As the patient did not improve even after the full course of Linezolid therapy for Methicillin
Resistant Staphylococcus aureus (MRSA) which was superadded infection, the discharge and also tissue material
were collected from the base of ulcer and cultured on Lowenstein – Jenesen media and Ziehl-Neelsen staining were
done. The acid fast bacilli were present on staining and growth on Lowenstein - Jenesen media was identified as
Mycobacterium tuberculosis.
Outcome of study: As the patient was also HIV positive both ATT and ART was started. A significant
improvement of the cutaneous lesion was noted after one month of treatment and patient was discharged after
another fifteen days.
Conclusion: Tuberculosis is very common in India and sophisticated automated system for detecting M.tuberculosis
is not available in all centers. Any non-healing ulcer not responding to routine antibiotics must be screened for
tuberculosis in developing countries. If tuberculosis is detected, promptly HIV testing must be done so that
treatment strategy can be finalised.
Word count - 341
EnglishHIV/TB co-infection, People living with HIV and AIDS (PLWHA), Skin tuberculosisINTRODUCTION Tuberculosis is one of the oldest disease, known to mankind and its evidence is detected even in Egyptian mummies. It remains as a leading cause of death worldwide, especially in developing countries like India, China etc. In the initial period of twentieth century, V.A.Moore has rightly said that as a destroyer of mankind, tuberculosis has no equal [1]. Almost one third of world population is infected with tuberculosis and in 1993 World Health Organisation (WHO) has declared tuberculosis an global emergency [2]. The risk of developing tuberculosis is estimated to be between 20-37 times greater in people living with HIV than among those without HIV infection[3]. Similarly, tuberculosis accelerates the progression of HIV infection to Acquired Immuno Deficiency Syndrome (AIDS) and shorten the survival of such patients. Of 1.8 million HIV related deaths in 2009, 22% were due to tuberculosis[4]. Even risk of drug resistant tuberculosis is higher amongst persons with HIV infection compared to others (HIV negative). Tuberculosis skin ulcers are extremely unusual. Cutaneous tuberculosis is caused by Mycobacterium tuberculosis and rarely by Mycobacterium bovis. Even in India and China where tuberculosis is quite common, cutaneous tuberculosis cases are rare i.e. 0.1 to 2.5%[5]. Moreover, Seeman et al in 2008 have reported that cutaneous tuberculosis is still a difficult disease to diagnose[6]. Here, we present a case with ulcerative lesion on right side of chest. The patient was diagnosed as a case of cutaneous tuberculosis with HIV and was treated with proper antiretroviral therapy and antitubercular drugs. Word count - 246 CASE STYDY OBSERVATIONS AND RESULTS: A 38 years old man presented with multiple ulcers over right axilla extending to the right chest wall with purulent discharge, was admitted to our hospital. The patient gave the history of small swelling over right axilla, 6-8 months back which was gradually increasing in size and was painful. As the patient was an agricultural worker, the history of trauma or thorn prick was specifically asked to rule out any actinomycotic or fungal infection. The patient had history of persistent cough 3-4 months back. But there was no history of weight loss. Patient was treated by many doctors from time to time but patient did not respond. The swelling was around 4cm × 3cm and as it was on the lower part of right axilla, it was diagnosed as axillary abscess and incision and drainage was done two and half month back, in a private nursing home. The pus was not sent for any investigation and patient was treated with antibiotic. As patient was very anaemic, 2 bottles of blood transfusion was given in the nursing home. Then other axillary swelling developed and ulcerated within one and half month. Hence, multiple ulcers with purulent discharge developed extending from right axilla to right chest wall( Photo 1). The investigations done in our hospital was: Fasting plasma glucose levels: 90 mg/dl, Haemoglobin: 6.1 gm/dl, ESR: 138 mm in first hour, Peripheral smear showed microcytic hypochromic anaemia, platelets were adequate and other parameters were in normal limits and X – ray chest: Lungs clear. Fine needle aspiration cytology (FNAC) from ulcerative lesion showed acute inflammatory cells. Gram?s staining of the pus showed plenty of Gram positive cocci arranged in clusters. On routine culture, Methicillin Resistant Staphylococcus aureus(MRSA) was isolated which was sensitive to Vancomycin and Linezolid and resistant to Penicillin, Erythromycin, Ciprofloxacin and Quinapristine, Dalfopristine. Methicillin resistance was detected using Cefoxitin (Cx 30μg) disk as per CLSI guideline[7].The patient was treated with Linezolid but the ulcers were not healing. Considering high ESR and as the ulcers were not healing, the discharge was collected from edge and base of the ulcer and Ziehl Neelsen staining with 20% H2SO4 was done. In the smear plenty of Acid fast bacilli (AFB) were present and some were beaded in appearance (Photo 2). On the same day, patient?s sputum sample were examined and it was negative for Acid fast bacilli. On next day morning, the induced sputum was collected and smear showed plenty of Acid fast bacilli (3+) and some were beaded in appearance (Photo 3). As the patient was AFB positive, patient?s serum was tested for HIV antibody and the patient was found to be HIV positive as per NACO guidelines [8], though the patient did not give any relevant history. The tissue collected from base of ulcer was homogenized and concentrated by Petroff?s method and the deposit was inoculated into two bottles of Lowenstein-Jensen media (L-J). Patient?s sputum was also inoculated into two bottles of L-J media after Petroff?s method. The rough, tough and buff coloured colonies of M.tuberculosis appear on L-J slant on fourth week from sputum and from the tissue growth appeared on sixth week on L-J media (Photo 4). The AFB staining from the growth revealed plenty of Acid fast bacilli and the growth was niacin positive. As the patient was HIV positive and sputum and exudate from ulcer was positive for Mycobacterium tuberculosis, Antiretroviral therapy (ART) and antitubercular drug regimen was also started on the same day. The patient responded to the treatment very well and after three and half months came for follow up when skin ulcers healed completely. DISCUSSION : Cutaneous tuberculosis is also an ancient disease and were described long before Robert Koch identified Mycobacterium tuberculosis in 1882. Laennec in 1826 first gave the description of cutaneous tuberculosis on his own prosector?s wart which developed after an injury while performing autopsy on a patient with spinal tuberculosis[9]. In 1886, Reil and Paltauf established that the wart was a tubercular lesion[10]. The clinical varieties of cutaneous tuberculosis can be divided into three broad groups – a) patients who were not previously exposed to M.tuberculosis, b) patients who were previously sensitized and c) tuberculids that develops a hypersensitive response of a tuberculosis focus elsewhere in the body. As previously sensitized hosts are very common in developing countries like India, lupus vulgaris is the most common variety of cutaneous tuberculosis reported from India, followed by TB verrucosa cutis and scrofuloderma[5]. No systematic survey for prevalence of cutaneous tuberculosis has been carried out in India. In one of the study it was found that cutaneous tuberculosis was associated with tuberculosis in other organs in 22.1% patients and the other organ most commonly involved were lungs. Even in the present case, the patient was having tubercular ulcer along with involvement of lungs. Most studies also reported that male are most commonly affected. Cutaneous tuberculosis sometimes has very diverse clinical presentation. The initial presentation may resemble a common bacterial infection or the ulcerative lesion may have superadded bacterial infection [11, 12]. In our case, initially patient had superadded infection with Methicillin Resistant Staphylococcus aureus(MRSA). After taking full course of Linezolid, the ulcer remained as it is and that made us to think for doing acid fast staining from the discharge. In culture, the growth was identified as M.tuberculosis. Currently the cause of skin ulcers may be vascular ulcers, squamous cell carcinoma, rodent ulcers, tubercular ulcers etc [13]. The differential diagnosis of cutaneous tuberculosis also includes infections with Mycobacterium ulcerans and Mycobacterium marium, Cutaneous anthrax, Cutaneous leishmaniasis, Sporotrichosis, Cat scratch disease due to Bartonella henselae etc [14]. In our case, as the patient was suffering from tuberculosis and HIV seropositive, the patient was treated with ART and ATT. In 2009, out of 1.7 million people died from tuberculosis. 4,00,000(24%) were among people living with HIV. Tuberculosis is also one of the leading cause of morbidity and mortality among PLWHA. Hence, WHO implemented collaborative HIV / Tuberculosis activities to decrease the burden of HIV / Tuberculosis coinfection. In patients with latent Tuberculosis infection, the risk of developing active diseases is several hundred folds higher among persons who acquire HIV. In 2007 it was reported from Brazil, 80% reduction in tuberculosis cases in HAART treated compared to ART naïve HIV infected person [15]. CONCLUSION: It has been observed that HIV epidemic continues to fuel TB epidemics and each increasing the morbidity and mortality of the other. WHO recommends the implementation of the Three I?s for HIV / Tuberculosis co-infection to reduce the burden of Tuberculosis among people living with HIV[16]. The three I?s are – i) Intensive tuberculosis case finding, ii) Isoniazid preventive therapy and iii) Infection control for tuberculosis. There is strong evidence that Antiretro-viral Therapy (ART) can lower a person?s viral load and restore the immune system and hence, significantly reduces HIV and Tuberculosis. WHO in 2011 recommends earlier ART at ≤350 CD4 count and immediate initiation of ART for all patients with HIV/TB co-infection irrespective of CD4 count[17]. Proper training and continuing medical education of health care workers is needed for early detection of cases with HIV/TB co-infection, so that WHO treatment strategies can be followed for a better outcome of the patient. Hence to conclude, in India, China and other developing countries, any non-healing skin ulcer, not responding to routine antibiotics, must be screened for tuberculosis and if positive, the patient must be screened for HIV. ACKNOWLEDGEMENT: Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors/ editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2315http://ijcrr.com/article_html.php?did=23151. Meyer JA Captain of All these Men of Death : Tuberculosis Historical Highlights. St. Louis. Warren H Green. 1977 2. Khatri GR, Frieden TR, Controlling tuberculosis in India. The New England Journal of Medicine 2002;347:1420-1425. 3. Guidelines for intensified tuberculosis case finding and isoniazid preventive therapy for people living with HIV in resourceconstrained setting. World Health Organization, 2011. 4. UNAIDS 2010 Global Report fact Sheet in World Health Organization HIV/TB Facts 2011;1-8. 5. Raman M. Cutaneous tuberculosis. Chapter 25. In Tuberculosis. Sharma SK, Mohan A editors. Jaypee Brothers Medical Publishers New Delhi, 2nd ed. 2009;384-396. 6. Seeman R., Trabonlsi R and Kanj S. Primary Mycobacterium tuberculosis complex cutaneous infection, Report of two cases and literature review. International Journal of Infectious Diseases 2008;12: 472-477. 7. Clinical and laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility testing 2006; 9th ed. CLSI document M2-M9 Wayne, PA: CLSI, 2006. 8. Detection of HIV infection Ch. 6, In: National AIDS Control Organisation, Guidelines on HIV testing, March 2007;38-53. 9. Laennec RTH, Traite de l?auscultation mediate et des maladies des pneumons et du Coeur Vol 1, Paris : Asselin and Cie ;1826 p 649-650. Quoted in Marmelzat WL Laennec and the prosector?s wart??. Arch Dermatol. 1962;86:122-124. 10. Riehl G, Paltauf R Tuberculosis verrucosa cutis. Eine bisher notch nitch beschriebne Form von Hauttuberenlose. Vjschn Derm Syph 1886; 13:19 Quoted in Most com Lupus vulgaris Raman M 11. Tappeiner G, Wolff K. Tuberculosis and other Mycobacterial infections. In Freedberg IM, Eisen AZ, Wolff K et al. eds. Dermatology in general Medicine, Vol 2, 5th edn. New Yark: McGraw – Hill, 1999:2274- 2292. 12. Beyt BE Jr.,Orbals DW, Santa Cruz DJ, et al. Cutaneous mycobacteriosis: analysis of 34 cases with a new classification of the disease. Medicine 1981;60: 95-109. 13. A Morrone, F Dassoni, MC Pajno et al Ulcers of the face and neck in women with pulmonary tuberculosis: presentation of a clinical case, Clinical case report, Rural and Remote Health, The International Electronic and Journal of Rural and Remote Health Research, Education Practice and Policy, 2010:10:1485-1489(Online).
14. GL Dandagi. Primary tuberculosis of skin – a nodular variant rare case report, Journal of Clinical and Diagnostic Research, 2010;4:3561-65. 15. Miranda A, Morgan M, Jamal L, Laserson K, Barreire D Silva G, Santos J et al Impact of antiretroviral therapy on the incidence of tuberculosis: the Brazilian experience, 1995- 2001, 2007;PLos ONE 2 e826. 16. Guidelines on intensified tuberculosis case finding and isoniazid preventive therapy for people living with HIV in resource constrained setting, WHO 2011 http://whqlibdoc.who.int/publications/2011/97 89241500708_eng.pdf 17. Guidelines on antiretroviral therapy for HIV infection in adults and adolescents. WHO 2010 http://whqlibdoc.who.int/publications/2010/97 89241599764_eng.pdf
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareINFECTION RISK CONTROL IN "COMPUTER RADIOGRAPHY IMAGING PLATE" IN DIAGNOSTIC
RADIOLOGY DEPARTMENT
English160163Suresh SukumarEnglish Sushil YadavEnglishThis study was carried out in order to establish whether infection control measures we?re being undertaken sufficiently on Computer Radiography Imaging plate (CRIP) used in Radio Diagnosis and Imaging Department of our medical college.CRIP is used to obtain the take compuerised radiographic image. This study involved the swabbing of a sample of CRIP used within different Areas of the department. Swabs were taken from the area on the corners and the centre of the plate. Each plate was firstly swabbed to determine the current level Of microorganism contamination (determination of baseline data) and then again after recommended cleaning. Comparisons were then made between the number of microorganisms? present (colony forming units/cm2) pre and post-cleaning at each location. All CRIP were found to be contaminated with microorganisms. Methylated spirit used in the practice of medicine, with water and soap is used to clean the CRIP was found to be significantly reduce the amount of microorganisms present. The results suggest that the All CRIP were not being cleaned sufficiently which has infection control implications for the department. In order for cross contamination to be kept to a minimum an effective infection control policy needs to be employed and this Should be to carry out regular cleaning
EnglishInfection control; computer Radiography Imaging Plate; Hospital acquired infections; Radiology Department;Introduction
Infection within healthcare has been in the news and has become a high priority recently, particularly as some infections are becoming harder to treat. The resistance of antibiotics and other antimicrobial agents have been reported on along with concerns regarding the rise of methicillin resistant Staphylococcus aureus (MRSA) (1). Staphylococcus aureus is one of the most common of all bacteria and can cause superficial infections of the skin and serious infections (2).Epidemic strains exist, which spread easily from person to person and can cause ward closure and disrupt hospital services (3). Infection control in hospitals is concerned with decontamination; this prevents microorganisms reaching a susceptible site in sufficient quantities to cause infection or potential harm to patients (4). Hospitals can become contaminated with organic matter and potentially infectious organisms and a safe environment can only be achieved by decontamination in the form of cleaning, disinfection and sterilization, breaking the chain of infection (5). A major reason for the importance of infection control is to prevent the occurrence of Nosocomial or Hospital Acquired Infections. These are infections that occur during a patient?s stay in hospital which were not present or incubating at the time of admission (2). In contrast to community acquired infections these infections usually occur as a result of pathogens taking advantage of patients whose normal defences against infection are contravened (2).
Aim
To establish whether CR plate can become contaminated with microorganisms and become a potential reservoir for cross infection and if simple, regular cleaning can significantly reduce this cross infection risk
Objectives
(1) To determine whether there is currently a detectable Presence of microorganisms on a sample of CRIP (2) To determine any presence of microorganisms after recommended cleaning (methylated spirit used in the practice of medicine with water and soap is used to clean the CRIP). (3) To evaluate the findings and make suggestions for future practice, including recommendations for re-audit.
Method
20 CRIP were swabbed, from Different areas within the Radiology department. These included general X-ray rooms for inpatients (room number 1, room number 2, and room number 3) and outpatient?s room. Randomisation of the sample was not practical, as it was necessary to ascertain data from each area. It was possible to swab all of CRIP from general X-ray rooms for in-patients (room number 1, room number 2, and rom number 3) and outpatients. Swabbing was carried out with Tryptic Soy Agar contact plates which are used to sample flat surfaces of equipment. They consist of a domed surface which is placed gently upon the area causing any microorganisms to be transferred onto the agar (10) (13). The plates were taken to the microbiology laboratory for culturing. The current level of microorganism contamination on the sample CRIP is known as baseline data. Following the collection of baseline data, each CRIP in the sample was cleaned according to recommended guidelines and swabbing was repeated. Data collected from this part of the audit was to identify a standard to compare to future practice and any future infection control carried out on CRIP After the data is collected the presence and absence of infection is mentioned as presence in CRIP and absence in CRIP.
Result
From this work all CRIP pre cleaning were contaminated with microorganisms. Table shows pre and post-cleaning results for location general X-ray rooms for in-patients (room number 1, room number 2, rom number 3) and out-patients x ray room. Post-cleaning data demonstrates that on most of the CRIP the number of colony forming units was reduced after cleaning. All plates were inspected by microbiology staff to identify the range of microorganisms present. Species of microorganisms found across the samples included most significantly Gram positive cocci in the form of Staphylococci Both coagulases positive and negative.
Discussion
Despite the fact that no Methicillin resistant strains exist was present upon the CRIP sampled, microorganism growth was found on all CRIP. This compares with other studies (6- 9) Coagulase-negative staphylococci are found as normal skin flora and include for example Staphylococci epidermis. These bacteria rarely cause infection (2, 4, 13). It has however recently been recognised that Staphylococci epidermis can be an important cause of HAIs as it produces an extracellular polysaccharide, a type of slime that enables it to adhere to plastics and metals(2,11). Staphylococcus aureus was also identified and is a coagulase Positive staphylococci. One-third of the population carry it on their skin or in their nose and throat asymptomatically (3). However, it is an important pyogenic pathogen, causing pus to form, which can cause a range of superficial infections of the skin if it penetrates the dermis such as septic spots, boils and abscesses and other more serious problems such as osteomyelitis, septicaemia and pneumonia (2,3,11, 13) It is also an important cause of HAIs, being responsible for around 40 to 50% of surgical wound infections and approximately 25% of blood stream infections(2,13), and is particularly capable of developing resistance to antibiotics. Methicillin resistant strains exist (MRSA), which are found in greatest abundance In the hospital setting as many patients receive antimicrobial therapy and are vulnerable to serious infection (2), it is also becoming recognised as an important pathogen due to its ability to colonise and cause infection of biomedical devices.1 Staphylococci released in skin scales will collect in dust and survive for long periods of time in the environment (2, 13). Swain and Flint on (7) compared the use of soap and water with alcohol wipes and phenolic disinfectant for the infection control of X-ray cassettes and concluded that all cleaning methods had a significant reduction in bacterial numbers. However, the alcohol wipes were found by the authors to be 100% effective, because of this and ease of use they were recommended as the cleaning method of choice. Another study shed doubt on The use of alcohol wipes, forensic tools were used to look for the presence of blood on seemingly clean cassettes and results suggested that if alcohol wipes were used universally to clean cassettes they are ineffective in cleaning any that are blood soiled (12). In our study use of methylated spirit used in the practice of medicine (especially for cleansing the skin before injections or before surgery) with water and soap is more effective method of cleaning the CRIP.
Conclusion
The results of the audit suggest that the CRIP were not cleaned effectively. Although the microorganisms identified are quite harmless in the majority of cases, all have the potential to be pathogenic when coming into contact with the variety of patients that present for examination in the Radiology department. This possibility is increased in cases where for example, there are damaged sites of skin such as wounds or cannula insertion sites (2). An effective infection control policy for the cleaning of CRIP should be established as an essential method to reduce cross contamination. It can be concluded that cleaning with methylated spirit used in the practice of medicine (especially for cleansing the skin before injections or before surgery) with water and soap is more effective method of cleaning CRIP can significantly reduce the number of microorganisms present and it should be carried out routinely.
ACKNOWLEDGEMENT
Author acknowledges the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors /publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. The author is highly thankful to the referees for their very constructive, valuable suggestions and useful technical comments, which led to a significant improvement of the paper.
Englishhttp://ijcrr.com/abstract.php?article_id=2316http://ijcrr.com/article_html.php?did=23161. National Audit Office. The Management and control of hospital Acquired infection in acute NHS trusts in England. London: National Audit Office; 2000.
2. Wilson J. In: Infection control in clinical practice. 3rd Ed.Edinburgh: Elsevier Limited; 2006.
3. Gould D, Brooker C. Applied Microbiology for Nurses. Basingstoke: Macmillan Press Limited; 2000
4. McCulloch J, editor. Infection control, science, management and practice. London: Whurr Publishers; 2000.
5. Horton R, Parker L. In: Informed infection control practice. 2nd ed. Edinburgh: Churchill Livingstone; 2002.
6. Fox M, Harvey J. An investigation of infection control for X-ray Cassettes in a Diagnostic Imaging Department. Radiography 2008; 14:306e11.
7. Swain JA, Flinton DM. X-ray cassettes: a potential crossinfection Risk? Journal of Diagnostic Radiography and Imaging 2000; 3:121e5.
8. Smith A, Lodge T. Can radiographic equipment be contaminated? By microorganisms to become a reservoir for cross Infection? Synergy 2004; Dec: 12e7.
9. Hodges A. Radiographic markers: friend or fomite? Radiologic Technology 2001; 73:183e5.
10. Booth C. Microbe monitoring. Cleanroom Technology 2006; Oct: 18e20.
11. Meers P, Sedgwick J, Worsley M. The microbiology and epidemiology of infection for health science students. London:Chapman and Hall; 1995
12. Study on blood contamination reveals disturbing results.Society of Radiographers. Available from: www.sor.org/members/snnarchive/SNRAug0 3p07.pdf; 2003 [accessed 16/04/09].
13. „„Do lead rubber aprons pose an infection risk??? Helen Boyle, Ruth M. Strudwick (2010).
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14TechnologySTRESS ANALYSIS OF SPUR GEAR USING FINITE ELEMENT METHOD - A REVIEW
English164171Sushil Kumar TiwariEnglish Upendra Kumar JoshiEnglishThe basic aim of this review paper is to provide the information for calculating the stresses of an involute gear in meshing. Gears are critical component in the rotating machinery. The researchers throughout the years had given various research methods such as Theoretical, Numerical and Experimental. We prefer the Theoretical and Numerical methods because Experimental testing can be expensive. This study says Finite Element Method is the best numerical solution for calculating gear stress.
EnglishSpur Gear, Stress Calculation, Bending Stress, Hertz Stress, Finite Element Method.INTRODUCTION
Gears are use to transmit power and motion from one shaft to another. Spur gear is cylindrical in form and has teeth, which are of involute form in most cases. The tooth surface elements are parallel to the gear axis. Each gear tooth may consider as a cantilever beam. When it transmits the load, it subjected to bending. The bending stress is highest at the fillet and can caused breakage or fatigue failure of tooth in root region, whereas contact stresses are on the both side of the tooth may causes Scoring Wear, Bending and Pitting fatigue.[1] Pitting fatigue is a compressive fatigue occurring at the point of maximum Hertzian stress. Gear stresses are affected by surface hardness, carbon content, metallurgical structure and lubrication condition. Gear tooth strength is the ability to resist tooth breakage and the ability to resist pitting is referred as durability.[1] To calculating bending stress, Lewis Formula is used, and for calculating contact stress Hertz equation can be applied in spur gear.[2]
BENDING STRESS (LEWIS FORMULA) [1]
A gear tooth is essentially a stubby cantilever beam. At the base of the beam, there is tensile stress on the loaded side and compressive stress on the opposite side. The ability of gear tooth to resist tooth breakage usually referred to as their „Beam Strength?. Wilfred Lewis accurately calculated this in 1893; he conceived the idea of inscribing a parabola of uniform strength inside a gear tooth.
In the centre of the band, there is a point of maximum compressive stress. Directly underneath this point, there is a maximum subsurface shear stress. Just ahead of the band of contact, there is a narrow band of the compression and just behind the band of contact; there is a narrow region of tensile stress. A bit of metal on the surface of a gear tooth goes through a cycle of compression and tension each time, when a mating gear tooth passes over it. If gear tooth loaded heavily enough, and there will usually be evidence of both surface cracks and plastic flow on the contacting surface, and there may be a rupturing of the metal due to surface shear stresses. The stress on the surface of gear teeth are usually determined by formula derived from the work of H. Hertz?s; and these stresses are termed as Hertz stress. Hertz determined the width of the contact band and the stress pattern when various geometric shapes were loaded against each other. Considering figure (5),
Above Hertz formula can be applied to Spur Gears quite easily by considering that the contact condition of gears are equivalent to those of cylinders having the same radius of curvature at the point of contact as the gear have. Ali Raad Hassan [8] presented formula for calculating Hertz equation for contact stresses in the pair of mating spur gear teeth. As figure (6)
Where and are the pitch radius of the Pinion and Gear respectively and is the pressure angle. Equations (2) and (3) are use for quick calculation of Bending and Contact stress of a pair of mating gear efficiently. American Gear Manufacture Association (AGMA) provides the standard equations for calculating bending and contact stresses for a pair of mating gear including various factors. Following literature review gives a lot of information about stress calculation based on Lewis formula, Hertz theory, AGMA/ANSI (American National Standards Institute) equations and Finite Element Method.
LITERATURE REVIEW
Shuting Li (2002) researcher, performed loaded tooth contact analysis of a three dimensional, thin-rimed gear by presenting a method that combines the Mathematical Programming Method (MPM) with the 3-dimentional finite element method and compare the results with experiment. and concluded that FEM with MPM efficiently calculates bending stress in gears.[3] Andrzej Kawalec (2006) author gives comparative study of tooth-root strength evaluation methods used within International Standard Organization (ISO) and AGMA standards then verifying them with Finite Element method. The results allow for a better understanding of existing limitation in the current standards applied in engineering practice as well as provide a basis for future improvement of gear standard.[4] Jose I. Pedrero (2007) researcher used a nonuniform model of load distribution along the line of contact. He says if the load is, assume to be uniformly distributed along the line of contact, simple equations given by the linear theory of elasticity and the Hertzian contact model are not good agreement with experimental results. In this paper a non uniform model has been considered for stress analysis at contact ratio 2 and 2.5, and concluded that,
1. The critical tooth-root stress corresponds to contact at the inner point of the outer interval of the two pair tooth contact, and teeth loaded 60% of total transmitted load.
2. The critical contact stress corresponds to either one of the three following condition.
3. Contact at the inner point of contact and teeth loaded with 25% of the total transmitted load.
4. Contact at the inner point of the two pair tooth contact and teeth loaded with 40% of the total transmitted load.
5. Contact at the outer point of the inner interval of the two pair tooth contact, and teeth loaded with 60% of the total transmitted load.[5]
Shuting Li (2007) researcher presented 3- Dimentional Finite Element Method to conduct Surface contact stress (SCS) and Root bending stress (RBS) calculations for a pair of spur gear with Machining Error (ME), Assembly Error (AE) and Tooth Modification(TM). Moreover, found that ME, AE and TM exerts great effects on SCS and RBS of the gear. Results obtained by International Standard Organization (ISO) and Japan Gear Manufacturing Association (JGMA) standards are comparing with result of Finite Element Analysis. [6] Shuting Li (2008) researcher investigates the effect of addendum modification on contact strength, bending strength and basic performance parameters of spur gear. A face contact model of teeth and MPM and 3D finite element methods are used together to conduct loaded tooth contact analysis. He concluded that Hertz formula is not exact enough for the contact stress calculation because in engagement position the contact point is for away from the pitch point, also Hertz formula cannot be use for contact stress calculation of the gear at Tip or Root contact. Contact stress and contact width are changed slightly if addendum becomes longer and number of teeth is not changed.[7] Ali Raad Hassan (2009) author is analysed contact stress between two spur gear teeth in different contact position, between 0º to 30º. Start with an angle of 0º and end at 30º with an interval of 3º with corresponding length of contact and contact ratio. and found that a highest value of contact stress is at beginning of contact and then starts reducing until it reaches the location of single tooth contact. Result obtained by finite element analysis is compare with theoretical value which is found at geometrical contact point.[8] Rubin D. Chacon (2010) researcher analysed the contact stress between spur gear teeth using a plane model and validate Hertz stress and AGMA contact stress with finite element contact stress. Then concluded that FEM is able to simulate contact stress in a pair of mating gear, the contact stress is highest at higher point on the involute and lower at a single pair of teeth. Assume the full load transmitted and minimal at the pitch point of contact.[9] Konstandinos G. Raptis, (2010) author performed photo-elasticity test to determine stresses in a pair of mating gear. Obtained experimental results were comparing with the theoretical value of maximum stress at gear tooth root (when tooth is loaded at their most unfavorable contact point i.e. highest point of single tooth contact). Then concluded that the result obtained by the applied method is in reasonable value whereas it rises with increasing number of teeth on the large gear.[10] Wei Yangang,(2010) author provided a theoretical procedure for obtaining maximum contact stress at different point in meshing under the light of Hertz formula. and concluded that when the number of teeth on small gear is smaller than a certain value, the maximum contact stress in the meshing process is not generated in the inner critical point in the single tooth meshing region of the small gear.[11] Xianzhang FENG (2011) author use a precise model in large-scale CAD software and define the stress and displacement field to determine the maximum equivalent stress and maximum displacement. By defining the quasi-static characteristic of finite element model, shows that model can accurately simulate the distribution of equivalent stress and displacement change in the process of teeth meshing. The results are well agree with the actual meshing law and not only verify the correctness of model but also expect to dynamic analysis of gear.[12] Ignacio Gonzlez-Peraz (2011) researcher developed a finite element model and validate in terms of contact area, pressure distribution and maximum contact pressure for those cases where the Hertz theory can be applied and provide partial crowning to the finite element model where Hertz theory do not work properly. And concluded that analysis results gives good agreement with Hertz theory for calculating maximum contact pressure, contact area, and deflection with minute errors.[13] Ali Kamil Jabur, (2011) author investigates the characteristics of an involute gear system including contact stress between a pair of the gear from 3-dimansional analysis and compared the result with experimental data. Experimental setup uses DC servomotor and mounting the strain gauge in the tooth of the gear made by Polyimide materials. and concluded that increasing the spur gear design parameters (number of teeth with module) leads to improvement in the tooth strength and increasing the thickness of critical section and makes it able to withstand higher load.[14] S. Sankar, (2011) author used circular root fillet instead of standard trochoidal root fillet in gear. and concluded that the tooth deflection in the circular root fillet is less when compared to the trochoidal root fillet, further there is appreciable reduction in bending stress and contact shear stress for circular root fillet design in comparison to that of trochoidal root fillet design.[15] Seok- Chul Hwang (2011) author compared the variation of contact stress during rotation with contact stress at Lowest Point of Single Tooth Contact (LPSTC) and concluded that gear design that consider the contact stress in a pair of mating gear is more severe than that of the AGMA standard.[16] Massimiliano Pau (2012) researcher performed a tooth contact analysis, and contact area, contact pressure using an ultrasonic experimental setup. The results provide the information about the size and shape of the nominal contact area and contact pressure distribution. He concluded that the ultrasonic method has good potential as an effective tool in investigating contact problem in gear.[17]
CONCLUSION
The above literature review presents that the Finite Element Method is widely used for stress analysis in a pair of gear. In addition, FEM software has been use for performing meshing simulation. Almost in all of the above cases, contact stress calculation and Bending Stress calculation play more significant role in the design of gear. This study shows that Hertz theory is the basis of contact stress calculation and Lewis formula is use for calculating bending stress in a pair of gear. Theoretical result obtained by Lewis formula and Hertz equation and results found by AGMA/ANSI equations are comparable with Finite Element Analysis of spur gear.
Englishhttp://ijcrr.com/abstract.php?article_id=2317http://ijcrr.com/article_html.php?did=23171. Darle W. Dudley, Practical Gear Design, McGraw-Hill Book Company, 1954
2. Peter R.N. Childs, Mechanical Design, Second edition, Elsevier ButterworthHeinemaan, 2004
3. Shuting Li, „Gear Contact Model and Loaded Tooth Contact Analysis of a ThreeDimensional Thin-Rimmed Gear? Journal of mechanical Design, ASME, 2002; vol.124/511.
4. Andrzej Kawalec, Jerzy Wiktor, Dariusz Ceglarek, „Comparative Analysis of Toothroot Strength Using ISO and AGMA Standard in Spur and Helical Gear With FEM-based Verification? Journal of mechanical Design, ASME, 2006; vol. 128/1141.
5. Jose I. Pedrero, Izaskun I.Vallejo, Miguel Pleguezuelos, „Calculation of Tooth Bending Strength and Surface Durability of High Transverse Contact Ratio Spur and Helical Gear Drives? Journal of mechanical Design, ASME, 2007; vol. 129/69.
6. Shuting Li, „Finite Element Analyses for Contact Strength and bending Strength of a pair of spur gears with Machining Errors, Assembly Errors and Tooth Modifications? Mechanism and Machine Theory, Elsevier, 2007; 42: 88-114.
7. Shuting Li, „Effect of Addendum on Contact Strength, Bending Strength and Basic Performance Parameters of a pair of Spur Gears? Mechanism and Machine Theory, Elsevier, 2008; 430: 1557-1584
8. Ali Raad Hassan,„Contact Stress Analysis of Spur Gear Teeth Pair? WASET 2009; 58
9. Rubin D. Chacon, Luis J. Adueza, „Analysis of Stress due to Contact between Spur Gears? Wseas.us, 2010
10. Konstandinos G. Raptis, Theodore N. Costopoulos „Rating of Spur Gear Strength using Photo elasticity and the Finite Element Method? American Journal of Engineering and Applied Sciences, 2010; 3(1): 222-231.
11. Wei Yangang, Zhang Xiujuan, Liu Yankui, „Theoretical research on the maximum Contact Stress of Involute spur Cylindrical Gear Pair in the External Meshing Process? IEEE,2010
12. Xianzhang FENG, „Analysis of field of Stress and Displacement in process of Meshing Gears? vol.5, 2011
13. Ignacio Gonzalez-Perez, Jose L. Iserte, Alfonso Fuentes, „Implementation of Hertz theory and validation a Finite Element Model for stress analysis of gear drives with localized bearing contact? Mechanism and Machine Theory, Elsevier, 2011; 46: 765-783
14. Ali Kamil Jebur, L.A.Khan, Y.Nath, „Numerical and Experimental Dynamic Contact of Rotating Spur Gear? Canadian Center of Science and Education, Modern Applied Science vol.5;2011
15. S. Sankar. Muthusamy Natraj, „Profile Modification- A Design approach for increasing the Tooth Strength in Spur Gear? International Journal of Advance Manufacturing Technology, Springer,2011; 55: 1-10.
16. Seok-Chul Hwang, Jin-hwan Lee, „Contact Stress Analysis for a pair of Mating Gears? Mathematics and computer modelling, Elsevier, 2011
17. Massimiliano Pau,Bruno leban, Antonio Baldi, Francesco Ginesu, „Experimental Contact pattern Analysis for a Gear-Rack system? Meccanica,Springer,2012; 47: 51-61
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareFORMULATION AND EVALUATION OF XANTHAN GUM BASED FLOATING TABLET OF TRAMADOL
HYDROCHLORIDE
English172180Somnath PatilEnglish Swati JagdaleEnglish Shailendra KelaEnglish Varsha DivekarEnglishThe aim of the present work was to prepare floating tablets of Tramadol HCl (TH) using xanthan gum as carrier. TH is a centrally acting oral analgesic that blocks pain through opoid receptor binding and inhibition of nor epinephrine and serotonin reuptake. It has elimination half-life of about 6 hrs. It has an oral bioavailability of about 70% which is suitable for developing gastro retentive floating drug delivery system. The formulations were prepared by varying the concentrations of xanthan gum and sodium bicarbonate. The tablets were prepared by direct compression technique. Xanthan gum and Hydroxy propyl methyl cellulose (HPMC) were used as alone and in combination as matrix forming agent and sodium bicarbonate for development of CO2. The prepared floating tablets were evaluated for tablet properties such as hardness, thickness, weight variation, floating property. In vitro dissolution was carried out for 8 hrs in 0.1N HCl at 37±0.50C using USP paddle type dissolution apparatus. It was noted that, all the prepared formulations had desired floating lag time and constantly floated on dissolution medium by maintaining the matrix integrity. The drug release from prepared tablets was found to vary with varying concentration of the polymer, xanthan gum and HPMC K4M. From the study it was concluded that floating drug delivery system can be prepared by using xanthan gum as a carrier.
EnglishFloating Tablet, HPMC K4M, in- vitro dissolution, Tramadol HCl, Xanthan gum.Introduction:
The real issue in the development of oral controlled release dosage forms is not just to prolong the delivery of drugs for more than 12 hours, but to prolong the presence of the dosage forms in the stomach or upper gastrointestinal (GI) tract until all the drug is released for the desire period of time [1] . Rapid GI transit could result in incomplete drug release from the drug delivery device in the absorption zone leading to diminished efficacy of the administered dose [2]. Several approaches are currently used to retain the dosage form in the stomach. These include bio adhesive systems [3]. swelling and expanding systems,[4? 5] floating drug delivery systems (FDDS),[6? 7] and other delayed gastric emptying devices.[8] FDDS, also called hydrodynamically balanced system, is an effective technology to prolong the gastric residence time in order to improve the bioavailability of the drug.[9] This technology is suitable for drugs with an absorption window in the stomach or in the upper part of the small intestine,[10] drugs acting locally in the stomach,[11] and for drugs that are poorly soluble or unstable in the intestinal fluid.[12] FDDS have a bulk density lower than the gastric fluid and thus remain buoyant in the stomach, without affecting the gastric emptying rate for a prolonged period of time. While the system is floating on the gastric contents, the drug is released slowly. Based on the mechanism of buoyancy, two distinctly different technologies, i.e. noneffervescent and effervescent systems, have been utilized in the development of FDDS. The effervescent system utilizes matrices prepared with swellable polymers and effervescent components, e.g. sodium bicarbonate and citric acid or stearic acid. The matrices are fabricated such that in the stomach carbon dioxide is liberated by the acidity of the gastric contents and is entrapped in the gellified hydrocolloid. This produces an upward motion of the dosage form and maintains its buoyancy. In noneffervescent FDDS, the drug is mixed with a gel?forming hydrocolloid, which swells on contact with the gastric fluid after oral administration and maintains relative integrity of shape and a bulk density of less than unity within an outer gelatinous barrier. The air trapped by the swollen polymer confers buoyancy to these dosage forms. [13? 14] TH is a centrally acting oral analgesic that blocks pain through opoid receptor binding and inhibition of nor epinephrineand serotonin reuptake. TH is having short plasma half life 6 hrs which is suitable for developing gastro retentive floating drug delivery system.
Materials and Methods:
Materials:
Tramadol hydrochloride was provided as a gift sample from JCPL, Jalgaon and hydroxy propyl methyl cellulose K4M and xanthan gum obtained as a gift sample from Vapi Care Pharma Pvt Ltd. Vapi. Other excipients and chemicals were of analytical grade and purchased from Pure Chem. Laboratories, Pune.
Method:
Floating tablets containing TH were prepared by direct compression technique using variable concentrations of Xanthan gum and HPMC K4M with sodium bicarbonate. Different tablets formulations were prepared by direct compression technique. All the powders were passed through 60 mesh sieve. Required quantity of drug, and low-density polymer were mixed thoroughly. Magnesium stearate was finally added as lubricant. The blend was directly compressed (9mm diameter punches) using tablet compression machine. Each tablet contained 100mg of TH and other pharmaceutical ingredients as listed in table-1. Table 1: Composition of Floating Tablet of TH
Evaluation of powder blend:
The powder blend used for preparation of tablets was evaluated for angle of repose, and compressibility index. Angle of repose: The angle of repose is a relatively simple technique for estimation of the flow property of a powder. Powders with low angle of repose are free flowing and those with a high angle of repose are poorly flowing powders [7]. 10 gm of powder was passed through funnel and the pile was formed. The height and weight of the pile was measured and the angle of repose was calculated by using the formula:- Angle of repose (θ) = tan-1 (height /radius) The angle of repose less than 300 usually indicate a free- flowing material and more than 400 suggest a poorly flowing material [8] . Carr`s compressibility index: The Carr?s compressibility index was calculated by calculating the tapped and bulk density using the 100 ml measuring cylinder. Compressibility is calculated by the formula. Carr`s compressibility index = (TBD-LBD)/ TBD X100 Where, TBD is tapped bulk density and LBD is loose bulk density. A carr`s index greater than 25 is considered to be an indication of poor flowability, and below 15, of excellent flowability. [9] Table 2: Characterization of precompresion parameters of table.
Evaluation of Tablets:
All the formulations were evaluated for various parameters such as hardness, friability, weight variation, % drug content, buoyancy lag time, swelling index, in-vitro drug release, release experiments, IR spectroscopy and optimized formulation were evaluated for in-vivo study. Hardness: Hardness of tablets was determined using monsanto hardness tester. Friability: For each formulation, the friability of 20 tablets was determined using the Roche friabilator. In this test tablets were subject to the combined effect of shock abrasion by utilizing a plastic chamber which revolves at a speed of 25 rpm, dropping the tablets to a distance of 6 inches in each revolution. A sample of pre weighted 20 tablets was placed in Roche friabilator which was then operated for 100 revolutions i.e. 4 mins. The tablets were then dusted and reweighed. Percent friability (%F) was calculated as follows, % F= (loss in weight / initial weight) x 100 Conventional compressed tablets that lose less than 0.5 to 1.0% of their weight are generally considered acceptable.
Thickness:
Thickness of all tablets was measured using a vernier calliper. Weight variation: The weight of 20 tablets was taken on electronic balance and the weight variation was calculated. The weight variation tolerance allowed for tablet weighing 324 mg and more is 5%.
Drug content:
To calculate the drug content, the tablets were triturated in the mortar. 30mg of the tablet powder (10mg Tramadol HCL) was added to 10 ml of distilled water and drug solution was filtered through Whatman paper no.1. The sample was analyzed for drug content by UV spectrophotometer (Varian Cary 100) at 270 nm after suitable dilutions. Drug stability in the 0.1 N HCl and distilled water was checked using UV spectrophotometer for a period of 10 hours. Buoyancy Studies: In vitro buoyancy was determined by buoyancy lag time. The tablets were placed in a 100 ml beaker containing 0.1N HCl. The time required for the tablet to rise to the surface and float was determined as floating lag time. Swelling index: The swelling index of the tablets was calculated in order to find out the swelling ability of the tablets. For calculating the swelling index, the previously weighed tablets were placed in the 100 mL beaker containing 0.1 N HCl. The tablets were removed at the time interval of 1 hr for 8 hours and weighed. The swelling index of the tablets can be measured by studying its dimensional changes, weight gain or water uptake. Hence swelling index was calculated by the formula; Swelling index= (Wt-Wo) X 100/Wo Where, Wt= Final weight of tablets at time„t? Wo= Initial Weight of tablets.
In Vitro Dissolution Studies:
The release rate of Tramadol HCl from floating tablets was determined using United States Pharmacopeia (USP) 24 dissolution testing apparatus 2 (paddle method). The dissolution test was performed using 900 mL of 0.1N HCl, at 37 ± 0.5°C and 60 rpm. A sample (5 mL) of the solution was withdrawn from the dissolution apparatus hourly for 10 hours, and the samples were replaced with fresh dissolution medium. The samples were filtered through a 0.45-μ membrane filter and diluted to a suitable concentration with 0.1N HCl. Absorbance of these solutions was measured at 270 nm using double beam UV spectroscopy. Cumulative percentage drug release was calculated using PCP disso software.
Results:
Evaluation of tablets:
Hardness:
Hardness of the formulations F1-F13 was observed within the range of 7.4-9.3 as shown in table 3.
Friability:
Friability of the tablets was observed below 0.30% for all batches which was in the acceptable limit. Thickness: The thickness of all the tablets was found within the range of 3 ± 0.5 mm. Weight variation: The weight of all the tablets was found within the range of 300 mg ± 5mg. Hence the weight of all formulations was found within the limit. Drug content: The range of % drug content of the formulations F1-F13 was found between 96.01 and 102.87. The tablets showed hardness, friability, thickness, weight variation and % drug content within the limit. The in-vitro buoyancy study shows that all the formulations shows good floating property.
In Vitro Buoyancy Studies:
The in-vitro buoyancy study showed the good floating ability of the tablets as shown in the table 3. Buoyancy lag time indicates the time required for the formulation to float in the medium. From table 2, it was observed that formulations F10 and F12 shows comparatively more floating lag time as compared to other formulations. It was further observed that formulation F2 shows lowest floating lag time among the all formulations.
Swelling index:
From the swelling index study of all the batches, it was observed that the increase in the concentration of polymers increases the swelling property of the tablets as shown in table 3. Further the formulation containing optimized swelling index was obtained. From the formulation batches, it was observed that the formulations F10 and F13 showed maximum swelling index. In Vitro Dissolution Studies: The drug release patterns from all the formulations are shown in table 3. The percent drug release after 8 hours is as shown in figure 1, 2 and 3. It was observed that the drug release was retarded from the formulations containing more concentration of Xanthan gum and HPMC K4M. The drug release profile of formulations F1-F13 indicates that as the concentration of polymer increases, the drug release was retarded. From the comparison of release profile of all the batches, it was observed that the formulations containing combination of polymers shows more retardation in drug release in less concentration as compared to Xanthan gum and HPMC K4M alone. This indicates that combination of polymers is more efficient in formulating the sustained release dosage form. Table 3: Evaluation results of formulations F1-F13 Fig.1: Effect of Various conc. of Xanthan gum on drug release Fig. 2: Effect of Various conc. of HPMC on drug release Fig. 3: Effect of Xanthan gum and HPMC in Various combinations on drug release
Discussion:
The flow properties of the powder blends were studied and formulation F5 was found to have comparatively good compressibility index and hausner ratio than other formulations. From the results of floating properties it was shown that all tablets had good floating properties. From the results of swelling study it was concluded that swelling index increases as time passes because the polymer gradually absorbed water due to hydrophilic in nature and swell. The swelling index increases with time up to 2 hours in some batches which might due to low viscosity of polymer and after 2 hours, the polymer chain relaxation was dominating phenomenon as swelling reaches thresholds resulting in lowering of swelling index. Thus the viscosity of polymer had a major role on swelling process, matrix integrity as well as floating capability. From the swelling study of the formulations, formulation F10 was found to have good swelling properties. The in vitro dissolution was carried out for all batches. From the dissolution studies of the formulations, formulation F10 was found to have better drug release profile than other formulations. Formulation F10 was also found to have better swelling capacity and floating time than other formulations. The drug release was extended for more than 8 hours from the floating drug delivery formulations.
Conclusion:
Regulated drug release attained in the current study indicates that the matrix tablets of Tramadol hydrochloride, prepared using various polymers, can successfully be employed as a once-a-day oral controlled release drug delivery system. High floating ability of the formulation is likely to increase its GI residence time, and eventually, improve the extent of bioavailability. However, appropriate balancing between various levels of the 2 polymers is imperative to acquire proper controlled release and flotation of the formulation. Formulation F10 shows good in vitro gastro retentive floating drug delivery of Tramadol HCl.
Acknowledgements:
Authors are thankful to JCPL, Jalgaon for providing the drug sample of Tramadol hydrochloride, also thankful to Vapi care pharma Ltd. for providing Xanthan gum and HPMC K4M. Authors are very much thankful to Principal and management of MAEER?s Maharashtra Institute of Pharmacy, Pune for their help and support. Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2318http://ijcrr.com/article_html.php?did=2318[1] Bardonnet P, Faivre V, Pugh WJ, Piffaretti JC, Falson F. [2006] Gastro retentive dosage forms: Overview and special case of helicobacter pylori. J. Control. Release 111:1 – 18.
[2] Obaidat AA, Obaidat RM. [2001] Controlled release of tramadol hydrochloride from matrices prepared using glyceryl behenate, Eur. J. Pharm. Sci. 52: 231-235
[3] Brunton L. Parker K. Blumenthal D. I. Buxton, Goodman and Gilman`s: Manual of pharmacology and therapeutics, Mc Graw Hill, New York. 358-367.
[4] Tiwari SB, Murthy TK, Pai MR, Mehta PR, Chowdhary PB. [2003] Controlled release formulation of Tramadol hydrochloride using hydrophilic and hydrophobic matrix system AAPS PharmSciTech 31: 1-6
[5] Narendra C, Srinath MS, Ganesh B. [2006] Optimization of bilayer floating tablet containing Metoprolol tartrate as a model drug for gastric retention. AAPS PharmSciTech 34:E1-E7.
[6] Dave BS, Amin AF, Patel MM. [2004] Gastroretentive drug delivery system of ranitidine hydrochloride: formulation and in vitro evaluation AAPS PharmSciTech 34: 1- 6.
[7] Aulton ME. [2008] Aulton`s Pharmaceutics-The design and manufacture of medicine, second ed., Churchill Livingstone Elsevier:. 133, 441-450.
[8] Lachman L, Lieberman HA, [2009] the theory and practice of industrial pharmacy, CBS publishers and distributors, special Indian edition, New Delhi. 300,301,317,299.
[9] Allen LV, Popovich NG, Ansel HC. [2008] Ansel`s pharmaceutical dosage form and drug delivery system, Lippincott William and Wilkins: 186-203.
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Table Caption:
1. Table 1: Composition of Floating Tablet of TH
2. Table 2: Characterization of precompresion parameters of table
3. Table 3: Evaluation results of formulations F1-F13
Figure Caption:
1. Fig.1: Effect of Various conc. of Xanthan gum on drug release
1. Fig. 2: Effect of Various conc. of HPMC on drug release
2. Fig. 3: Effect of Xanthan gum and HPMC in Various combinations on drug release
Table 1: Composition of Floating Tablet of TH
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareVIBRATION THRESHOLD OF UPPER LIMB DURING ULNT1 IN INDIVIDUALS WITH TYPE II DIABETES MELLITUS AND NON DIABETIC INDIVIDUALS
English181187Mamta MohanEnglish Ravi Shankar ReddyEnglish Ganesh BMEnglishBackground: Type II diabetes mellitus patients have been shown to affect the multimodal sensory, reflex and motor systems in distal extremities. Studies have examined the mechanosensitivity and vibration threshold in type II diabetes mellitus patients in the lower limb and compared it with normal individuals. There is scanty literature available in comparison of the vibration threshold in the upper limb in type II diabetes mellitus patients with non diabetic individuals. Methods: Thirty type II diabetic individuals were included in the diabetic group and thirty asymptomatic age matched individuals were taken to match the group. Vibration threshold (VT) was measured by tester 1 at the baseline for both the groups using a bioesthesiometer capable of deriving a vibration of 100 Hz. After the VT was taken at three levels 1) at the baseline, the tester performed the upper limb neurodynamic test 1(ULNT1) for each individual. During the sequence of the ULNT1, 2) vibration threshold was measured at initial onset of pain i.e. P1 and 3) short of maximal pain i.e. P2. Results: Repeated measures ANOVA was used to compare the VT differences within group and between groups. There was a statistical significant difference between the vibration threshold of diabetic and non diabetic group at the three levels with a p value < 0.001. Conclusion: Vibration threshold of the upper limb is higher in individuals with type II diabetes mellitus as
compared to non diabetic individuals.
EnglishVibration threshold, ULNT1, Diabetes mellitusINTRODUCTION
Diabetes mellitus is a group of metabolic diseases characterized by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. Type II diabetes mellitus is most common form which is a disease of insulin resistance that usually has relative (rather than absolute) insulin deficiency1 . Earliest change in diabetic nerve function is alteration in axonal excitability due to changes in ion conductance of axon membrane due to metabolic processes directly affecting the nerves, microvascular abnormalities of the endoneurium and auto immune inflammation. Four main mechanisms have been postulated to underlie the pathogenesis of nerve pathology in diabetes mellitus, which are metabolic processes directly affecting nerve fibres, endoneurial microvascular disease, autoimmune inflammation and deranged neurotrophic support.2 It is due to these effects of hyperglycemia that peripheral nerve involvement is highly frequent in type II diabetes mellitus and it has been documented that one third of type II diabetic patients have peripheral neuropathy3 . Among the nerves, there is a tendency of the large diameter nerve fibers that mediate sense of vibration to get involved first in diabetes mellitus.2 Neurodynamic tests involve sequential limb movements that are employed to include the link between mechanical and physiological types of mechanisms. An aim of using these tests in assessment of a nerve is to stimulate mechanically and move neural tissues in order to gain an impression of their mobility and sensitivity to mechanical stresses so as to evoke the physiological responses.4 In order to assess the upper limb nerve function, the standard upper limb neuro dynamic test 1 (ULNT1) is usually used as it evokes symptoms of distribution of the median nerve because the forces generated by the test are biased towards this structure.4 There are various techniques of assessing the conductivity of nerve such as NCV that basically assesses the motor and sensory aspects of the nerve whereas the vibration threshold (VT) reflects particular function of the peripheral nervous system especially the somatosensory pathway.5 Type II diabetes mellitus patients have been shown to affect the multimodal sensory, reflex and motor systems in distal extremities. Mechanosensitivity in diabetes mellitus patients should be considered as an essential inclusion in the assessment to predict the extent of involvement of the nerve.6 Studies have also been done to determine the vibration threshold in lower limb in normal individuals but there is scanty literature available in comparison of the vibration threshold in the upper limb in type II diabetes mellitus patients with non diabetic individuals.
METHODOLOGY
Study Design
This study took place in department of physiotherapy, Manipal University, Manipal, India. A cross –sectional 2-group design was used. Completion of questionnaires and all measurement procedures were conducted in the same room on each occasion.
Subject Selection
Type II diabetic subjects in the study were selected from all patients presenting for the first time to physiotherapy outpatient and inpatient clinic over a one year period. 30 subjects with mean age of 55.60 ± 9.79 were included in the study. All new patients completed a simple questionnaire as part of the inclusion-exclusion procedure. On daily review of these first stage questionnaires, the clinical records of patients who provisionally met the inclusion criteria were subjected to secondary detailed screening by an experienced member of the physiotherapy faculty who is experienced the in the field of musculoskeletal physiotherapy and diabetic neuropathy. The Diabetic subjects with clinical signs of neuropathy were excluded from the study. After this screening, subjects who met inclusion criteria invited to participate in the study and were given further verbal and written information about the study, and were asked to read and sign a consent form. For controlled age matched normal subjects an advertisement was given in physiotherapy department and Manipal University for their voluntary participation in the study. To be considered for inclusion, the subjects must have been aged between 30 to 70 years, have had no history of diabetes, upper limb disorders, Cervicobrachial pain syndrome, Acute inflammatory/ demyelinating diseases, Any recent surgeries in upper limb. Finally, eligible 30 control subjects were selected by age to ensure a similar distribution to the patient group. The mean age of the subjects was 53.43±9.96. The subjects first session were to familiarize them with the equipment and vibration threshold testing tasks. All participants signed a written consent form prior to participating in the experiment. Ethics approval was obtained from the Manipal University Ethics Committee.
Measurement of Vibration Threshold (VT)
VT was measured by tester 1 at the baseline for both the groups using a bioesthesiometer capable of delivering a vibration of 100 Hz. The subjects were made to sit comfortably on a chair with hand and arm completely on the pillow. The probe of the Vibrometer was placed at the pulp of the distal phalanx of the thumb.7 Either right or left hand was tested. The subjects were shielded from the Vibrometer display during testing. At baseline, tester 1 first increased the vibration to a point where the subject perceived the stimulus. This was taken as appearance of vibration. Then the intensity was further increased and slowly reduced till they identified the disappearance of the stimulus. This measurement was done thrice and the average of the six values was taken as the vibration threshold. After the VT was taken at the baseline, the tester performed the ULNT1 (adopted from M.Shacklock)6 for each individual. For this a pressure biofeedback inflated to 50 mm Hg was used to prevent shoulder elevation. Then the shoulder was abducted to 90-110 degrees followed by complete external rotation, forearm supination, wrist and finger extension. The last component of ULNT1 was elbow extension and elbow extension value was recorded using universal goniometer as a measure of mechanosensitivity. During the sequence of the ULNT1, the occurrence of the first response of elbow extension i.e. pain considered as P1 was noted. The angle of its occurrence was measured with the universal Goniometer and VT at this position in the same manner as that of baseline was taken for both the groups by the tester 2. The next occurrence of the symptom i.e. P2 at which any further movement was intolerable was noted. The corresponding elbow extension angle of P2 was measured. The range of elbow extension was reduced until the feeling of discomfort disappeared and VT was measured at this point for both the groups by the tester 2. The reduction of elbow extension was adopted to avoid the masking of pain for perception of vibration. The measurement of vibration threshold was measured for both diabetic individuals and age matched normal individuals.
Data Analysis
The statistical analysis was done using the SPSS 14.0 for Windows software. The statistical significance value was set at 0.05 with 95% confidence interval and p value less than or equal to 0.05 would be considered as significant. Repeated measures ANOVA were used to compare the VT differences within group and between groups.
Results
Demographic data regarding the age (yrs), sex and duration of individuals with type II diabetes mellitus and non diabetic individuals are shown in table no. 1 Analysis of repeated measures ANOVA shows the comparison of the vibration threshold between the diabetic and the non diabetic group at 3 levels i.e. VT at baseline, VT at P1 and VT at less than P2. There was a statistical significant difference between the vibration threshold of diabetic and non diabetic group at the three levels with a p value < 0.001. This states that the vibration threshold was found to be raised in the diabetic group at all the three levels compared to the non diabetic group. There was no statistical significant difference between the three levels within each group with a p value of 0.755.This states that there was no difference in vibration threshold during the ULNT1 procedure within each group (Table no. 2 and Figure 1) Thus vibration threshold of the upper limb is higher in individuals with type II diabetes mellitus as compared to non diabetic individuals. However, vibration threshold did not change within subjects of each group during the ULNT1 testing.
Discussion
Our study aimed at comparing the vibration threshold of the upper limb during ULNT1 in individuals with type II diabetes mellitus and non diabetic individuals. As per the results, the vibration threshold was found to be increased in the individuals with type II diabetes mellitus as compared to the non diabetic individuals. Vibration threshold is a measure of conductivity i.e. a function of the axon in conducting the impulse from the external receptor. Thus, alteration of the vibration threshold in type II diabetic individuals may be due various reasons. Studies in human and animal models with diabetes mellitus have shown reduced nerve perfusion and endoneurial hypoxia. Investigations on biopsy materials from patients with mild to severe neuropathy show graded structural changes in nerve microvasculature including basement membrane thickening, pericyte degeneration and endothelial cell hyperplasia. Arterio-venous shunting also contributes to the reduced endoneurial perfusion. These vascular changes strongly correlate with clinical defects and nerve pathology. Early vasa nervorum functional changes are caused by metabolic insults of diabetes, the balance between vasodilator and vasoconstrictor are altered.8 The findings of our study regarding the vibration threshold are in contrast with the study done by David A Gebler et al. They conducted a study to check the vibratory and thermal thresholds in normal and diabetic individuals using an Optacon Tactile Tester (OTT) and Thermal Sensitivity Tester. They did not find the vibratory and thermal threshold of diabetic subjects to be different from the normal individuals. But in diabetic individuals with neuropathy, the thermal and vibratory thresholds were found to be increased.9 .
Conclusion
Vibratory threshold of the upper limb in type II diabetic individuals is higher than the non diabetic individuals. Vibration threshold and Mechanosensitivity in diabetes mellitus patients should be considered as an essential inclusion in the assessment to predict the extent of involvement of the nerve.
Englishhttp://ijcrr.com/abstract.php?article_id=2319http://ijcrr.com/article_html.php?did=23191. Fazan V, Vasconcelos C, Valença M, Nessler R, Moore K. Diabetic Peripheral Neuropathies: A Morphometric Overview. Int J Morphol 2010;28:51-64.
2. Raymond AA. Management of Diabetic Neuropathy. Malaysian Journal of Medical Sciences 2003;10:27-30.
3. Comi G, Corbo M. Metabolic neuropathies. Curr Opin Neurol 1998;11:523-9.
4. Shacklock M. Clinical Neurodynamics - A new system of musculoskeletal treatment. London: Elsiever 2005.63-65
5. Lise H, Jepsen JR, Sjogaard G. Vibrotactile sense in patients with different upper limb disorders compared with a control group. Int Arch Occ Env Hea 2006;79:593-601.
6. Boyd BS, Wanek L, Gray AT, Topp KS . Mechanosensitivity during lower extremity neurodynamic testing is diminished in individuals with Type 2 Diabetes Mellitus and peripheral neuropathy: a cross sectional study. BMC Neurology 2010;10:75.
7. Colette R, Jane G, Nicola J. P. Effect of straight leg raise examination and treatment on vibration thresholds in the lower limb: a pilot study in asymptomatic subjects. Man Ther 2005;10:136-143.
8. Cameron NE, Eaton SE. Vascular factors and metabolic interactions in the pathogenesis of diabetic neuropathy. Diabetologia 2001;44:1973-1988.
9. Gelber DA, Pfeifer MA, Broadstone VL, Munster EW, Peterson M, Arezzo JC. Components of variance for vibratory and thermal threshold testing in normal and diabetic subjects. J Diabetes Complications 1995;9:170-176.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241417EnglishN2012September14HealthcareCardiovascular Co-Morbidity and role of exercise in Rheumatoid Arthritis: A Review
English188194Shahnawaz AnwerEnglish Ameed EquebalEnglishPatients with rheumatoid arthritis have an increased risk of cardiovascular disease (CVD). Cardiovascular event rates are markedly increased in rheumatoid arthritis (RA). Data from population- and clinic-based epidemiologic studies of rheumatoid arthritis patients suggest that individuals with rheumatoid arthritis are at risk for developing clinically evident congestive heart failure (CHF). The vasculature plays a crucial role in inflammation, angiogenesis, and atherosclerosis associated with the pathogenesis of inflammatory rheumatic diseases. There is overwhelming evidence that, in the general population and several at risk subpopulations, exercise provides significant physical and psychosocial benefits, and facilitates management and improvements of outcome in Rheumatic disease. There is increased recognition of the need for structured preventive strategies to reduce the risk of CVD in patients with RA. In this review, the research agenda for understanding and preventing CVD comorbidity in patients with rheumatoid arthritis is discussed.
EnglishRheumatoid arthritis, Cardiovascular Disease, Inflammation, ExerciseIntroduction:
Cardiovascular disease is an increasingly recognized contributor to excess morbidity and mortality in rheumatoid arthritis (RA) [1-3]. The most probable cause of cardiac death in rheumatoid arthritis, as in the general population, is atherosclerotic coronary artery disease leading to ischaemic heart disease [4] . Rheumatoid arthritis, which is characterized by inflammatory polyarthritis with progressive joint damage, occurs in about 0.5%-1% of adult population in most countries [5]. Several studies have shown a higher incidence or prevalence of ischaemic cardiac pathologies such as myocardial infarction, congestive heart failure, and coronary deaths in patients with rheumatoid arthritis than in the general population [3,6-7] . Traditional cardiovascular risk factors do not adequately account for the extent of cardiovascular disease in RA [3,8]. Although hypertension and age are potential additional contributors to cardiovascular events in this disease [6], markers of current and cumulative inflammation (white cell counts and radiographic joint damage, respectively) are associated with ultrasonographically determined subclinical atherosclerosis [9] – a predictor of cardiovascular events [10] . The use of methotrexate is associated with a significantly lower risk for cardiovascular (CV) events in RA patients compared with patients who had never used disease-modifying antirheumatic drugs (DMARDs) [11]. Suissa and colleagues [12] found a negative association between the rate of myocardial infarction and the current use of any DMARD in a case control study. A study from Sweden [13] suggested that the risk for developing first CV events in RA was lower in patients who were treated with tumour necrosis factor-alpha (TNF-α) blockers. In this review we examine the evidence of risk for CVD in patients with rheumatoid arthritis and the suggested underlying mechanism and discuss the role of exercise for the prevention and management of CVD in such patients.
Epidemiology of Cardiovascular disease in patients with rheumatoid arthritis
Fewer statistics on incidence and prevalence rates for Congestive heart failure (CHF) in patients with RA are available and are derived from a handful of population-based[7,14] and clinic-based RA cohorts[15]. Gabriel et al. [7] estimated the incidence of CHF among all RA patients in Olmsted Country, Minnesota, from data abstracted from medical records. Between 1955 and 1985, 78 cases of incident CHF were identified among 450 prevalent cases of RA compared to 54 cases among the same number of non-RA community controls matched for age, sex, and baseline comorbidity, yielding a relative risk of 1.60 (95% CI 1.12-2.27). In contrast, the risk of incident CHF in patients with osteoarthritis (OA), a noninflammatory arthritis, was not increased compared to non-OA community controls [7] . In a follow-up retrospective review of the same cohort extended to 1995, now using the Framingham diagnostic criteria for CHF, Nicola et al. [14] confirmed an increased risk of incident CHF in both rheumatoid factor (RF) negative and positive RA patients compared to non-RA controls adjusted for age, sex, and CV risk factors. Incident CHF risk remained elevated after further adjustment for comorbid ischemic heart disease, although the risk relationship was no longer statistically significant for RF negative patients in this model [14] . In a combined cohort of RA patients from community-based practices and drug safety monitoring studies (n = 9093), Wolfe et al. [2] estimated an adjusted lifetime relative risk of CHF in patients with RA is more as compared with OA controls. The adjusted lifetime prevalence of CHF in the RA population was 0.7 % greater as compared to OA controls. Data were collected via patient survey of self reported, physiciandiagnosed CHF, and confirmed by review of a random sample of medical records in 50% of patients reporting CVD events. In a subsequent analysis [15], in which the drug safety cohort represented a third (n = 4,307) of the total sample (n = 13,171), Wolfe et al. reported an adjusted frequency of CHF of 3.9% (95% CI 3.4-4.3%) in RA patients compared to 2.3% (95% CI 1.6-3.3%) in controls with knee or hip OA. Factors associated with prevalent and incident of CHF were those typically associated with CHF in the non-RA population (e.g., age, male gender, hypertension, coronary artery disease, diabetes, and smoking) while RA-related measures (patientreported disability, pain, and RA global severity) were also associated with prevalent and incident of CHF.
Risk factors for cardiovascular events in patients with rheumatoid arthritis:
Traditional risk factors for vascular disease such as, smoking, hypertension, diabetes and hyper lipidemia are important for the increased risk of CVD in subjects with RA [16]. However, traditional risk factors alone do not fully explain the excess CVD risk in RA. Other non-traditional factors are hypothesized to play a role, in particular the burden of inflammation as indicated by the Creactive protein (CRP) and/or erythrocyte sedimentation rate (ESR) [17, 18]. In a communitybased cohort of patients with inflammatory polyarthritis, Goodson, et al also noted that excess CVD mortality was confined to patients who were rheumatoid factor positive [19]. These markers of inflammation and inflammatory burden confer additional risk of CVD death in those with RA after adjusting for traditional CVD risk factors and comorbidities [20] . Severity of disease has consistently been associated with an increased risk of CVD events in RA. Patient with severe extra articular RA manifestations are at an increased risk of developing coronary artery disease [21] as well as peripheral vascular disease [22]; and severe extra articular RA is a predictor of both overall mortality [23] and cardiovascular mortality [20] , indicating that systemic inflammation is a major determinants of vascular comorbidity in RA. In contrast with the general population, a low body mass index (BMI), rather than obesity, has been associated with increased CVD in patients with RA [24, 25] .
Role of exercise in Rheumatoid arthritis and Cardiovascular disease:
Exercise is one of the most important behavioral interventions that can have a major beneficial impact on the likelihood to develop, suffer symptomatically or die from CVD. Any physical activity is better than no, or little, physical activity. There is overwhelming evidence that, in the general population and several at risk subpopulations, exercise provides significant physical and psychosocial benefits, and facilitates management and improvements of outcome in Rheumatic disease. It helps maintain a healthy life-style, reduce CVD risk factors including obesity [26], dyslipidaemia [27, 28], hypertension [29] , diabetes mellitus [30] and possibly even inflammation [31]; it is also effective for preventing acute coronary syndromes [32-34]. Moreover, exercise helps the management of established CVD: both aerobic exercise [35, 36] and resistance training [37] improve myocardial contractility and quality of life in patients with chronic heart failure and produce significant functional benefits in people with intermittent claudication [38]. More importantly, cardiac exercise rehabilitation programmes are an important part in the management of patients after an acute coronary syndrome (ACS) [39] and lead to significantly improved quality of life and reduced mortality rates [40, 41] . The overall physiological adaptations that occur as a result of exercise [42] provide protection against CVD mortality, even in the presence of well-established CVD risk factors [43, 44]. CVD mortality is lower in highly fit than in moderately fit individuals [45], while physical inactivity is an independent risk factor for the development of CVD [46, 47]. Even though cardiorespiratory fitness may have a familial component [48], it can be increased significantly by exercise training, regardless of age, gender, race and initial fitness levels [49]. The required activity levels can be accrued through formal training programmes or leisure-time physical activities [50]. Moreover, supervised exercise programmes are more effective compared with non-supervised exercise [51, 52], most likely due to greater adherence. Great controversy still exists about the optimum amount of exercise for eliciting the greatest cardiovascular benefit. Different exercise intensity [53] and duration [54], as well as various combinations of them [55], may have different impacts on the magnitude of cardiorespiratory fitness improvement. Most authors agree that there is a dose–response relation between the amount of exercise, all-cause and cardiovascular mortality [53, 54]. The greatest potential for reduced mortality is in sedentary individuals (such as many RA patients), in whom even slight increases in daily physical activity are beneficial [56,57]; for more active individuals, higher levels of intensity should be pursued [56]. Depending primarily on the starting levels of physical activity, cardiovascular fitness has been reported to increase by 8–51% following an exercise intervention [54, 56] . Moderate-intensity exercise of long duration appears to elicit the most benefit on CVD risk and mortality [53, 55-57]. Current guidelines by the American College of Sports Medicine (ACSM) suggest that an individual should engage in exercise at least three times a week, at an intensity of 60–80% of maximum oxygen uptake (VO2max), for at least 20–30 min, in order to experience significant improvements in cardiorespiratory fitness and optimum cardiovascular benefits [58]. In terms of caloric expenditure, this can be translated to 1000–2000 kcal/week [59]. These calories can be expended in either continuous exercise or accumulated from several short bouts of exercise during a day [59, 60] . Aerobic exercise is the most appropriate, but this can be supplemented by low-to-moderate intensity resistance training [60]. The exercise regimen should be reconsidered regularly, usually every 4– 6 weeks, based on the principles of exercise periodization [61] so that participants continue to improve their performance.
Summary
There is strong evidence that persons with rheumatoid arthritis are at high risk for developing cardiovascular disease. Several studies have shown a higher incidence or prevalence of ischaemic cardiac pathologies such as myocardial infarction, congestive heart failure, and coronary deaths in patients with rheumatoid arthritis than in the general population. Severity of disease has consistently been associated with an increased risk of CVD events in RA. CVD mortality was more confined to patients who were rheumatoid factor positive. Role of exercise in the management and prevention of CVD in RA patients is very important yet neglected area of RA patients treatment programmes. As this review shows, there is accumulating evidence that in patients with RA, exercise therapy is effective in improving the prognostic risk factor profile. There is little has been investigated and published on the role of exercise as a means to control risk and manage CVD in individuals with RA. More research is required to identify the optimal regimens, timing and environment for exercise, as well as educational and behavioural intervention that will facilitate long term adherence to an active life style and/or structured exercise.
Acknowledgments:
We acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. We are also grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2320http://ijcrr.com/article_html.php?did=23201. Solomon DH, Karlson EW, Rimm EB, Cannuscio CC, Mandl LA, Manson JE, et al. Cardiovascular morbidity and mortality in women diagnosed with rheumatoid arthritis. Circulation 2003;107:1303-07.
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8. Dessein PH, Joffe BI, Stanwix AE: Inflammation, insulin resistance, and aberrant lipid metabolism as cardiovascular risk factors in rheumatoid arthritis. J Rheumatol 2003;30:1403-05.
9. Kumeda Y, Inaba M, Goto H, Nagata M, Henmi Y, Furumitsu Y et al. Increased thickness of the arterial intima-media detected by ultrasonography in patients with rheumatoid arthritis. Arthritis Rheum 2002;46:1489-97.
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11. Van Halm VP, Nurmohamed MT, Twisk JWR, Dijkmans BAC, Voskuyl AE. Diseasemodifying antirheumatic drugs are associated with a reduced risk for cardiovascular disease in patients with rheumatoid arthritis: a case control study. Arthritis Res Ther 2006;8:R151.
12. Suissa S, Bernatsky S, Hudson M. Antirheumatic drug use and the risk of acute myocardial infarction. Arthritis Rheum 2006;55:531-6.
13. Jacobsson LT, Turesson C, Gulfe A, Kapetanovic MC, Petersson IF, Saxne T et al. Treatment with tumor necrosis factor blockers is associated with a lower incidence of first cardiovascular events in patients with rheumatoid arthritis. J Rheumatol 2005;32:1213-1218.
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