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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesBIOCONTROL AND BIOREMEDIATION POTENTIALS OF PSEUDOMONAS ASSOCIATED WITH RHIZOSPHERE REGIONS OF SUNFLOWER (HELIANTHUS ANNUUS L)
English0109Raval A.A.English Desai P.B.EnglishBackground and Aims: To combat the extensive use of chemical pesticides and other agrichemicals on plant growth which may be hazardous to soil and in turn human health, a group of rhizobacteria- the Plant Growth-Promoting Bacteria- can be employed. This study focuses on the mechanisms exhibited by these rhizobacteria that can further help to promote plant growth. Methods: Antifungal activities of different Pseudomonas isolated from sunflower (Helianthus annuus L.) rhizosphere were tested against four phytopathogenic fungi. Study of other biocontroling factors like production of Hydrocyanic acid (HCN) and Ammonia were also taken into consideration. The other study was pertaining to the production of several lytic and detoxification enzymes that these bacteria possess. Tolerance of these isolates to heavy metal stress was also found out, as heavy metal pollution is one of the major problems of environmental concern. Important Findings:
Results indicated that 30% of isolates inhibited the growth of Fusarium, Aspergillus and Helminthosporium, 50% were antagonistic against Curvularia. Among the other biocontrolling factors studied, two isolates showed presence of HCN and Ammonia. Study of hydrolytic enzyme showed that protease production was observed in five isolates, one produced amylase. Three isolates produced lipase and cellulase enzymes. Two of the isolates tolerated about 120 mM concentration of Chromium and three isolates grew in presence of 9.9 mM concentration of Zinc.
EnglishRhizosphere, Plant growth-promoting rhizobacteria, Biocontrol, Antifungal, Agrichemicals.INTRODUCTION
In the Bio-Era of today the rhizobacteria are playing a major role in various aspects to sustain agricultural development. Among these, the plant growth promoting rhizobacteria (PGPR) [1] are the beneficial ones that help to stimulate and enhance plant growth[2]. These bacteria exhibit a wide array of potentials for plant growth promotion. They directly benefit the plant by providing them with phytohormone indole acetic acid, Gibberellic acid, cytokinins etc., solubilizing minerals like Phosphorous and providing Nitrogen. Indirectly the PGPR may act as biocontrolling agents[3] , suppress plant diseases by secreting antifungal metabolites that support plant growth. May they be antibiotics[4], other volatile compounds, ironchelating siderophores, HCN[5], Ammonia[6] , Hydrolytic enzymes[7]. Here the study is focused on an important group of PGPR, the Pseudomonas, which have good potentials of Biocontrol[8] and tolerance to specific amount of heavy metals Lead, Chromium (VI)[9] and zinc.. The Helianthus annuus L. plant in association with these rhizobacteria can help improve phytoremediation[10] and rhizoremediation. Different agricultural plants assimilate and accumulate heavy metals[11] in a specific manner depending on a variety of factors controlling the soil plant system. Microorganisms are an important element in this system. The use of such associations in the rhizosphere may help in improving the structure of derelict and contaminated soils. Sunflower is one of the important oil seed crops in India. India has fourth largest area under Sunflower cultivation in the world and accounts for 10% of world acreage. Its share in total world production is about 5%. However the yield at 570 kg/ha is lowest among the major Sunflower producing countries in the world[12] . The seeds of Sunflower may be subjected to attack by different soil borne and seed borne fungi. The fungi can hinder seed germination and seed quality of Sunflower. Use of PGPR may thus help in controlling such seed borne diseases in these plants. Some common fungi associated with all the Sunflower varieties are Alternaria tenuis (32- 35%), Aspergillus flavus (0.5-47%), Aspergillus niger (8.7-40%). Species of Aspergillus, Fusarium, Alternaria and Penicillium have also been recorded on stored seeds [13] .
MATERIALS AND METHODS
Sample Collection: An intact plant Helianthus annuus L. was uprooted with a surface sterilized garden trowel and placed in a sterile plastic bag along with the adhering soil. Three samples were collected. Non rhizosphere soil was collected and suspension was prepared in sterile 0.1 M MgSO4 to disperse the microorganisms [14] . Root systems from the experimental plants with adhering soil were used to isolate rhizosphere microorganisms. Root segments of the plant were shaken vigorously for 3-5 min. in a test tube of sterile 0.1 M MgSO4 to dislodge the adhering soil and attendant microorganisms [2]. To isolate the endophytic bacteria excised surface sterilized roots were macerated with a sterile mortar and pestle diluted in PBS (Phosphate Buffer Solution), containing 20% glycerol. The suspensions were plated onto specialized media for Pseudomonas, King’s B medium. Plates were then incubated at ambient temperature after which growth of bacteria allowed differentiation by morphological characteristics of colonies. All three types of samples were processed and analyzed as follows. Isolation and characterization of isolates: Colony morphology was observed on the specialized media. Then the bacterial strains were characterized by morphological, biochemical and physiological tests including pigment production, gram reaction, catalase and oxidase production, nitrate reduction activities by standard microbiological procedures. Ten different bacterial strains were selected and characterized. Then the bacterial strains were assessed for the antifungal activity. 1. To assess the ability of the bacterial strains to inhibit fungi, each isolate was tested against four different phytopathogenic fungi with the circle method. Briefly bacterial isolates were seeded in a 9 cm diameter circle on a PDA plate. After 24 hr at room temperature, a 5 mm plug of each fungus was placed on the centre of the circle. Plates were incubated at 28° C for 3 days. Plates without bacterial strains were used as control. Fungal growth inhibition was assessed by measuring the mycelial radial growth [3] . 2. Testing of these bacteria in liquid media was also assessed in dual culture. The growth medium for in vitro test of fungal antagonism was sterile yeast extract sucrose-mineral salts broth (YESM). Twenty ml of sterile YESM broth was dispensed into sterile 9 cm glass petri plates. The procedure used in screening for in vitro antagonism of Pseudomonas strains against Fusarium was as follows. One ml bacterial suspension (2.5 x 106 CFU/mL) prepared from a 24 hr culture of each strain was dispensed into 20 mL sterile YESM broth in 9 cm glass petri plates and incubated for 48 hr at ambient temperature for growth of bacteria, after which three 5-mm agar plugs cut from 3 day-old culture of Fusarium were placed in the centre of each plate and further incubated in the dark for three days. Cultures were frequently agitated during incubation for proper aeration of the medium. Control treatment consisted of YESM containing three agar plugs of the fungus but without bacteria. Mycelia of the fungus were collected after three days by passing the culture through weighed Whatman No. 1 filter paper. Then washed twice with 50 mL deionized water. The harvested mycelia on the filter paper were oven dried over night at 60° C. In vitro antagonism of each strain was assessed by comparing the mycelium dry weight of the fungus in the control treatment with that of the fungus in the fungus-bacteria dual culture [14] . Then HCN production by the isolates was detected by the method of Bakker & Schippers [15]. Freshly grown cells were spread on King’s B medium amended with 4.4 g/L Glycine. A sterilized filter paper saturated with 0.5 % solution of picric acid and 2% sodium carbonate was placed in the upper lead of the petri dish. The petri dish was then sealed with parafilm and incubated at 30° C for 4 days. A change in colour of the filter paper from yellow to reddish brown as and index of cyanogenic activity was recorded. An uninoculated control was kept for comparison of results [16] . Production of Ammonia: Production of ammonia was determined by the method of Dye, 1962 by growing the isolates in peptone water in tubes and detecting it by adding Nesslar’s reagent in each tube [17] . The enzymatic activity such as protease and amylase were determined as described by Smibert & Kreig, 1994. Hydrolysis of Tween 20 was recorded as white precipitation around the colonies [18] . Production of cellulase was determined in M9 medium [19] amended with yeast extract and carboxymethyl cellulose. After 8 days incubation at 28º C, isolates surrounded by clear halos were considered positive. Alternatively the plates were flooded with Congo Red solution for 15 minutes then rinsed with 1 M NaCl for 10-15 mins. Clear zones were considered positive. Pectinase production was determined in the same M9 medium with pectin (4.8 g/L). After 2 days of incubation at 28º C, the plates were flooded with 2 M HCL and isolates surrounded by clear halos were considered positive for pectinase production [3] . Heavy metal tolerance: The bacterial strains were grown in a nutrient medium with different concentrations of heavy metals to assess their tolerance to these metals. Metals like Chromium, Zinc and Lead were analyzed. Organisms were grown in increasing concentration of the metals Zn+2, Cr (IV) and Pb+2 and the broth was incubated at 37° C for 48 hr. Bacterial growth was monitored by optical density measurement (OD 660 nm). Maximum resistance level was determined [20] .
RESULTS
A total of ten different strains of bacteria were isolated associated with Sunflower rhizosphere regions. Isolates S1 to S4 were bulk soil isolates, isolates R1, R2, R3 were rhizosphere isolates and isolates ER1, ER2 and ER3 were isolated from endorhizosphere regions. The strains were recovered on King’s B medium. The data presented in table 1 shows the colonial characteristics. Isolates S1, S2, R1, R2 and R3 produced diffusible and nondiffusible pigments of yellow to bluish green colour. The strains were identified from their biochemical, morphological and nutritional characteristics. Assessment of the antagonistic activity of the isolates showed that four of the strains showed more than 50% inhibition of Fusarium. Isolate R1 & ER1 showed 64% inhibition against this fungus. Two of the rhizosphere isolates R1 & R2 showed 75% and 70% inhibition respectively against Aspergillus niger and 56% inhibition was seen in isolate S2. Isolates R1, R2 and R3 showed 73% inhibition against Curvularia species. Isolate S1 and R3 showed 50% inhibition against Helminthosporium and isolate R1, R2 and ER3 showed 75% inhibition against this fungus as compared to controls (Table 2, Figure 1). Dual liquid culture method which was carried out by dry weight determination also showed 48% inhibition of growth of Fusarium in isolate R1 and 36% inhibition of growth in case of isolate R2. From the data presented strain identified as Pseudomonas aeruginosa (R1) was inhibitory against all the four phytopathogenic fungi tested. These two strains also produced other biocontroling factors like the hydrolytic enzymes, protease, lipase and volatile metabolites HCN and Ammonia (Table 3). The tolerance of the isolates towards heavy metals like Chromium, Zinc and Lead is as shown in the table 4 and 5. Pseudomonas aeruginosa and Pseudomonas flourescens tolerated 60 mM and 120 mM concentration of Chromium, 2.2 mM and 4.4 mM concentration of Zinc respectively. 9.9 mM concentration of Lead was tolerated by both of them (Table 6). These data are in accordance with several authors [21] .
DISCUSSION
The adverse effects of pesticides and other agrochemicals on the environment and human health have generated interest in searching for alternative ways to control pests and diseases in plants. It is more crucial for developing countries like India. Biological control of plant diseases involve the use of antagonists selected by testing their ability in vitro and in vivo [2]. Here the study was carried out on the free living strains of rhizobacteria specifically associated with and important oil crop, the Sunflower (Helianthus annuus L.). The bacteria which are native to the crop and therefore can be more effective as biocontroling agents and may indirectly help plant growth promotion. Treatment of seeds with such bioagents i.e. Pseudomonas flourescens gave superior results and was at par with chemicals like triadimefom (12.23 q/ha), Carboxin (12.54 q/ha), Carbendazin (13.03 q/ha) and Captan (12.59 q/ha) [23]. Again, taking into consideration the bioremediation aspects, the selected strains were also able to tolerate specific concentrations of heavy metals tested. As pollution by heavy metals is posing a problem to the environment Chromium in higher concentration may prove toxigenic and carcinogenic [21] Chromium toxicity includes poor growth and leaf chlorosis [22]. Lead is also classified as a ‘possible human carcinogen ‘by IARC which lead to lung cancer, stomach cancer and gliomas [24]. Such bacteria possessing multiple capacities of biocontroling and bioremediation capabilities can be effectively used for plant growth promotion as plant growth promoting rhizobacteria (PGPR). Such strains when used could be able to survive for carrying out its plant growth promoting functions in soils containing high concentration of these metal ions.
ACKNOWLEDGEMENT
I am grateful to the management Shri Bhartiya Vidya Mandal, The Principal Arts, Science & Commerce College, for their co-operation and support. I also acknowledge my sincere thanks to The Director, SRICEAS, for providing me excellent research facilities.
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2. Kloepper JW, Hume DJ, Scher FM, Singleton C, Tipping B, Laliberte M et al. Plant growthpromoting rhizobacteria on Canola (rapeseed). Plant disease. 1988; 72: 42-46.
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4. Keel C, Schnider U, Maurhofer M, Voisard C, Laville J, Burger U et al. Suppression of root diseases by Pseudomonas flourescens CHAO: importance of bacterial secondary metabolite, 2, 4-diacetylphloroglucinol. Mol PlantMicrobe Interact 1992; 5: 4-11.
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7. Compant S, Duffy B, Nowak J, Clement C and Barka EA. Use of plant growth promoting bacteria for biocontrol of plant diseases: Principles, Mechanisms of Action and Future Prospects.
8. Johri BN, Rao CVS, Goel R. Flourescent Pseudomonads in plant disease management. In: Dadarwal KR (ed.) Biotechnological approaches in soil microorganisms for sustainable crop production. Jodhpur, India: Scientific Publishers, 1997; 193-221.
9. Faisal M and Hasnain S. Plant growth promotion by Brevibacterium under Chromium stress. Research Journal of Botany. 2006; 1(1): 24-29
10. Zadeh BM, Firozabadi GRS, Alikhani HA and Hosseini HM. Effect of Sunflower and Amaranthus culture and Application of Inoculants on Phytoremediation of the Soils Contanimated with Cadmium. AmericanEurasian J. Agric. Environ. Sci., 2008; 4(1): 93-103
11. Tahsin N, Hristeva T and Masheva V. Productivity and Rhizosphere microflora of different Sunflower Genotypes grown in an industrially polluted region. Journal of Environmental Protection and Ecology 4, 2003; No. 1, 39-43.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareA PROSPECTIVE RANDOMIZED CONTROL STUDY COMPARING RUBBER BAND LIGATION WITH INJECTION SCLEROTHERAPY FOR GRADE II HAEMORRHOIDS
English1023Brahadeeswaran.SEnglish Tirou Aroul.TEnglish Robinson Smile.SEnglishBackground: Various modalities of treatment for haemorrhoids were attempted but the best treatment remains unanswered despite the wide variety of treatment options in use. Although surgical haemorrhoidectomy is a more definitive procedure, it has a reputation of having a significant postoperative pain and an extended recovery time. Hence non-surgical procedures such as Rubber band ligation, injection sclerotherapy, cryotherapy, etc. are becoming popular nowadays. Aim: This prospective study to compare the results of rubber band ligation and injection sclerotherapy in grade II haemorrhoids. Methods: A Prospective cross sectional study of 60 patients attending the outpatient department of general surgery in MGMC & RI with complaints suggestive of haemorrhoids were undertaken from September 2009 to June 2011. DRE & Proctoscopy done for all these patients to identify the grade of haemorrhoids. All patients were randomized into group B (Rubber band ligation) and group S (Injection sclerotherapy) with 30 patients in each group. Procedure was performed and patients were observed immediately following the procedure and at 2 weeks, 1 month, 3 month, 6 months and 1 year. Results: In group B, it was uneventful when observed immediately with p < 0.001 where as in group S, bleeding and pain were noticed with p < 0.001 and p = 0.024 respectively. At 2 weeks follow up, pile mass was sloughed out in group B and fibrosed in group S without any significant difference. At the end of 1 year, all 30(100%) patients in group B and 26(86.7%) patients in group S were symptom free with p = 0.038.Conclusion: Thus the outcome of both Rubber band ligation and Injection sclerotherapy were the same in treating grade II haemorrhoids except for the fact that the immediate post procedure complications like bleeding and pain were significantly more in patients who had Injection sclerotherapy
EnglishHaemorrhoids, Rubber band ligation, injection sclerotherapy, pain, bleeding.INTRODUCTION
Haemorrhoid is one of the commonest surgical diseases1 . Haemorrhoids are frequently found on routine rectal examination in patients who have never had any complaints related to the disease. They are merely prolapsed mucosa with congested venous plexus. About 75% of people will have haemorrhoids at some point in their lives2 .Haemorrhoids are most common among adults ages between 45 to 653 . The terms ‘haemorrhoids’ and ‘piles’ are used quite commonly but they have entirely different meaning. The term haemorrhoid is derived from Greek which means bleeding (haima = blood: rhoos =flowing).The term pile is derived from the Latin word PILA which means ‘a ball’4 . The frequencies of II degree haemorrhoids are more common than other types. There are various modalities of managing haemorrhoids, both surgical and non- surgical such as rubber band ligation, injection sclerotherapy, cryotherapy, laser ablation, haemorrhoidectomy, stapled haemorrhoidopexy etc., Barron introduced Rubber band ligation in 1963. It has proved over the years to be the most successful, non-operative, method of treating I, II and III degree haemorrhoids. The success rate for band ligation reaches 78% of cases; the rest would need a second line of treatment5 . It has always been a relatively difficult procedure because of the equipment used. It needed two people, one to hold the Proctoscope and the operator to apply the bands using two instruments: one to draw the tissue and the other to fire the band. The instrument needed reloading of the bands after each application to each haemorrhoid. Newer instruments provided a use of suction to draw the tissue, so that the operator uses one hand to hold the proctoscope and the other to apply the suction and fire the band. The difficulty in stretching the rubber band to load the firing mechanism of the instrument continued to be a major one. The difficulty in application of rubber band resulted in introduction of other methods such as cryosurgery, laser treatment, infrared coagulation and radio frequency coagulation678 . In case of injection sclerotherapy, sclerosant is injected using Gabriel syringe or using a spinal needle over the pedicle of the pile mass. Common sclerosants used are Sodium Tetra Decyl Sulphate, 5% phenol, ethanolamine etc. The purpose of the present study is to compare rubber band ligation with injection sclerotherapy in grade II haemorrhoids in terms of resolution, recurrence and complication. AIM OF THE STUDY To compare the results of rubber band ligation and injection sclerotherapy in grade II haemorrhoids As regards. 1. Resolution 2. Recurrence 3. Complication
MATERIALS AND METHODS
A prospective cross sectional study of 60 patients attending the general surgery OPD in Mahatma Gandhi Medical College with complaints of bleeding and mass descending per rectum were undertaken from September 2009 – June 2011
All age groups of both sexes were included.
DRE (Digital Rectal Examination) and proctoscopy done for all these patients to identify the grade of haemorrhoids.
In this study group, II degree haemorrhoids were taken into account. Much attention was focussed on selecting only uncomplicated haemorrhoids.
The inclusion and exclusion criteria are
Inclusion criteria:
Grade II haemorrhoids
Exclusion criteria:
• Grade I, III & IV haemorrhoids.
• Thrombosed haemorrhoids.
• Infected haemorrhoids.
• Associated with malignancy.
• Inflammatory diseases of bowel.
• Bleeding diathesis.
• Patient who wants to undergo surgery.
Two different time tested methods of treatment for haemorrhoids were selected for comparative study. These procedures are preferred on the basis of
Compliance of the patient.
Cost factor. ? Simplicity of techniques.
Treated as OPD basis.
Maintenance of normal anatomy as far as possible to preserve the normal anal sensation.
Good curative rate.
Patient was well informed about both types of treatment and was randomized by lots.
B – Rubber band ligation in grade II
S – Injection sclerotherapy in grade II
This study was conducted after getting clearance from ethical committee.
a) Rubber band ligation
• Patient was prepared pre-operatively by glycerine enema.
• Rubber band placed in the Barron’s banding gun as shown in the figure A.
• Rectum packed with 2% lignocaine soaked gauze after the procedure. b) Injection sclerotherapy • DRE was done to assess the tone of the sphincter muscles. • 1ml of sclerosant (sodium tetra decyl sulphate) was taken in a syringe and injected over the pedicle of the pile mass using spinal needle (24G)
• Rectum packed with 2% lignocaine soaked gauze after the procedure. Patients of both groups were treated with ? T.Paracetamol 500mg x 3 days ? Laxatives: Syp. Liquid paraffin 15ml HS x 3 days & sos ? Dietary modification (High fibre diet, plenty of oral fluids)
Patients were reviewed after 2 weeks / 1 month / 3 months / 6 months / 1 year
Statistical Analysis
Comparison of proportion by ‘CHI SQUARE’ test
Comparison of mean value by student ‘T’ test
Study Parameters
Age
Sex
Grade of haemorrhoids
DRE & Proctoscopy findings
Type of treatment given
Bleeding
Pain
Mucous discharge
Recurrence
Others
OBSERVATION AND RESULTS
This prospective study was done to compare the results and outcome of Rubber band ligation with injection sclerotherapy in grade II haemorrhoids who presented to general surgery OPD in Mahatma Gandhi Medical College and Research Institute at puducherry. In our study, 60 patients who were diagnosed as grade II haemorrhoids were included, among them 30 patients were randomly selected in each group( B&S).The age group varied between 21 and 75 years of age with a mean of 46.27 in group B and 45.70 in group S (p =0.858) as shown in table 1 and fig 1 This study showed male predominance over female, 76.7% and 23.3% in group B, where as 86.7% and 13.3% in group S (p = 0.317) as shown in table 2 and fig 2 Among the 60 patients, 7(23.3%) patients presented with mass descending per rectum and 23 (76.7%) patients had bleeding along with mass descending per rectum as chief complaints in group B. In group S, 5(16.7%)patients presented with mass descending per rectum and 25 (83.3%)patients had bleeding along with mass descending per rectum with p = 0.519 (table 3 & fig 3) DRE revealed normal mucosa/tone in all 30 patients (100%) in group B where as in group S, 27 (90 %) patients had normal tone/mucosa and 3 patients (10 %) had sphincter spasm (table 4 &fig 4) All patients were observed immediately after the procedure and they are reviewed after 2 weeks, 1 month, 3 months, 6 months and 1 year. All patients were observed immediately after the procedure. In group B(Band Ligation), 28(93.3%) patients were comfortable in the absence of any complication with significant p < 0.001 where as in group S(Sclerotherapy), 10(33.3%) patients were comfortable. In group S, 20 (66.7%) patients had some complication with a significant p < 0.001. Out of 20 patients, 10 (33.3%) patients had bleeding with a significant p < 0.001. 6 (20%) patients had pain with a significant p = 0.024 and both in 4 (13.3%) patients with p =0.112 where as in group B, only 2(6.7%) patients had bleeding. (Table 5 & fig 5)
Follow Up
At 2 weeks of follow up, all patients were observed. Pile mass sloughed out in 29(96.7%) patients (p = 0.052) and 1(3.3%) patient had bleeding in group B. In group S, Pile mass got fibrosed in 23(76.7%) patients with p = 0.052, bleeding present in 4(13.3%) patients with p =0.195, mucous discharge present in 1(3.3%) patient and pain in 2(6.7%) patients. (Table 6& fig 6) At 1 month follow up in group B, 27(90%) were comfortable in the absence of any symptoms with significant p = 0.004. In group S, fibrosed piles mass seen in 6 (20%) patients with significant p = 0.010 and 5(16.7%) patients had bleeding with p = 0.448. 1(3.3%) had pain and one patient had recurrence with p = 1.000 as shown in table 7 and fig 7
At 3 months follow up, 30(100%) patients in group B were comfortable without any symptoms. In group S, 28(93.3%) patients were symptom free, 1(3.3%) had bleeding and one patient had recurrence with p = 1.000 (table 8 & fig 8) At 6 months follow up, 30(100%) patients in group B were comfortable without any symptoms. In group S, 28(96.6%) patients were comfortable. 1(3.4%) had recurrence with p = 1.000 and one patient had lost to follow up (table 9 & fig 9) At 1 year follow up, all patients were observed. 30(100%) patients in group B and 26(86.7%) in group S were comfortable without any symptoms with moderately significant p = 0.038.In group S, 3 (10%) patients had recurrence (p = 0.237) and 1 (3.3%) patients lost to follow up (table 10 & fig 10)
DISCUSSION
Over the years, various modalities of treatment for haemorrhoids were attempted but the best treatment remains unanswered despite the wide variety of treatment options in use. Safety is of paramount importance, especially when treating a benign disease such as haemorrhoids. 9 10 Although surgical haemorrhoidectomy is more definitive procedure, it has a reputation of having a significant postoperative pain and an extended recovery time.11 Nowadays, rubber band ligation is the most widely used procedure, and it offers the possibility to resolve haemorrhoidal disease without the need for hospitalization or anaesthesia, and with a lower incidence of complications when compared to conventional surgery.121314 Rubber band ligation can be used to treat grade I, II and III haemorrhoids and it is safe, effective and easy to use.15Other non-surgical treatment modalities include injection sclerotherapy, cryosurgery, infra-red photocoagulation, out of which injection sclerotherapy is widely used for its simplicity, cost effective and efficacy. Shamimqureshi et al conducted a study at Department of General Surgery, Jinnah Post Graduate Medical Centre at Karachi from January 2001 to May 2008. All Patients with II and III degree haemorrhoids were included in this study. In this study, haemorrhoids were more commonly found in males (74.88%) than in females (25.11%)16. In our study also males dominated with 81.6% than females with 18.4%. Patients were counselled regarding prospects of success of both the procedures. This study concluded that Rubber band ligation is a safe, effective and economical procedure for treating 2nd and 3rd degree haemorrhoids as outpatient basis which is similar to the present study in grade II haemorrhoids only. A comparative analysis was made by johnF et al regarding various non-surgical treatment such as rubber band ligation, sclerotherapy and infra-red coagulation in haemorrhoids. Rubber band ligation demonstrated a 25% difference with a response rate of banding being significantly better (p = 0.001) than sclerotherapy but the frequency of pain and bleeding are similar following both techniques17. In the present study, the response to treatment in both techniques were similar (p = 0.052) but bleeding (p < 0.001) and pain (p = 0.024) were present in patients immediately after injection sclerotherapy with a significant difference. GörgülAhmet et al conducted a study at Gazi University Faculty of Medicine at Ankara in which short-term results of rubber band ligation and injection sclerotherapy for II degree haemorrhoids are retrospectively evaluated and compared in a non-selected Turkish patient group. Thirty-four adult patients with II degree haemorrhoids are reported. All patients were supplemented by high fibre diet one week prior to the procedure. Rubber band ligation was applied to 18 patients and sclerotherapy to 16 patients. Both symptomatic status and objective healing criteria were evaluated. During a follow-up period exceeding 6 months (6-10 months), the success rate of Rubber band ligation with high-fibre diet was 88.9% (16/18), while it was 68.8% (11/16) in the Sclerotherapy group18. In our study, the success rate after one year is 100% in rubber band ligation group and 86.7% in sclerotherapy group. Both the methods in their study were free from notable complications where as in the present study, sclerotherapy group had immediate complications like bleeding (p Englishhttp://ijcrr.com/abstract.php?article_id=1991http://ijcrr.com/article_html.php?did=19911. Barron J2. Office ligation of internal Haemorrhoids. Am j surg 1963; 105: 563-673.
2. Baker H, Longe JL.Haemorrhoids. Gale Encyclopedia of Medicine 2006; 3: 1766–69.
3. Chong PS, Bartolo DCC. Haemorrhoids and fissure in ano. GastroenterolClin North Am 2008; 37:627–44.
4. Bailey and Love’s Short practice of surgery – 24th edition.
5. Finninger P. Modern treatment for Internal Haemorrhoids.Br Med J 1997; 314: 1211.
6. Liebach JR, Cerda JJ. Haemorrhoids: modern treatment methods. Hosp Med 1991 Aug: 53- 68.
7. Norman DA, Newton R, Nicholas GV. Direct current electrotherapy of internal haemorrhoids: an effective, safe and painless outpatient approach.Am J Gastroenteral 1989; 84: 482.
8. Templeton JL, Spence RJ, Kennedy TL, et al. Comparison of infra-red coagulation and rubber band ligation for 1st and 2nd degree haemorrhoids.Am J Gastrenteral 1989; 84: 475-81.
9. BatL,Meltzer E, Koler M, Dreznick Z, Shemesh E.Complication of rubber band ligation of symptomatic internal haemorrhoids. Dis Colon Rectum. 1993; 36: 287-90.
10. Bleday R, Pena JP, Rothenberger DA, Goldberg SM, Buls JG. Symptomatic haemorrhoids:current incidence and complications of operative therapy. Dis Colon Rectum 1992; 35: 477-81.
11. MacRae HM, McLeod RS. Comparison of haemorrhoidal treatments: a meta-analysis. Dis colon rect 1995; 38: 687-94.
12. Shanmugam V, Thaha MA, Rabindranath KS, KL. Campbell KL, Steele RJC, Loudon MA. Systematic review of randomized trials comparing rubber band ligation with excisional haemorrhoidectomy. Br J Surg 2005; 92: 1481-7.
13. Pezzullo A, Palladino E.Rubber band ligation of haemorrhoids:Five years follow up. Min Chir 2000; 21: 253-6.
14. Longman RJ, Thomson WH. A prospective study of outcome from rubber band ligation of piles. Colorectal Dis 2006; 8: 145-8.
15. Ayman El Nakeeb, Amir Fikry et al.Rubber band ligation for 550 patients of symptomatic haemorrhoids out of 2200 patients retrospective study. Egyptian J Surg 2008 Jul; 27(3): 125-31.
16. Shamimqureshi et al. Rubber band ligation of symptomatic internal haemorrhoids; result of 450 cases. Pak J Surg 2009 Jan-Mar; 14(1): 19-22.
17. JohnF et al. A comparative analysis of non surgical treatment of haemorrhoids. Am J gasteroenterol 1992; 87(11): 1601-5.
18. GörgülAhmet et al. The results and comparison of rubber band ligation and injection sclerotherapy supplemented by highfibre diet in the treatment of second-degree internal haemorrhoids. Tur J Gasteroenterol 1999; 10(1): 66-71.
19. Savioz.D, Roche.B, Glauser.T, Dobrinov.A,Ludwig.C, Marti.M.C. Rubber band ligation of haemorrhoids: relapse as a function of time. Int J Colorectal Dis 1998; 13: 154-56.
20. Wani NA, Fazilli A, Bhat GH, Mir IS.A Retrospective Study and Evaluation of Rubber Band Ligation as a Treatment of Haemorrhoids. Ind J prac doctors 2009: 5(6).
21. Watson NFS, Liptrott S, Maxwell-Armstrong CA. A Prospective audit of early pain and patient satisfaction following outpatient band ligation of Haemorrhoids. Ann R CollSurg Engl 2006; 88:275-79.
22. Kanellos I et al.A comparison of simultaneous application of sclerotherapy and rubber band ligation, with sclerotherapy and rubber band ligation applied separately for the treatment of haemorrhoids: a prospective randomized trial. Colorectal dis 2003 Mar; 5(2): 133-8
SIGNIFICANT FIGURES
+ Suggestive significance (P value: 0.05 * Moderately significant ( P value:0.01
** Strongly significant (P value : PRadiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareDEVELOPMENT OF ASSAY METHOD FOR ARTESUNATE BY INDIRECT SPECTROPHOTOMETRY OF ITS ALKALINE DEGRADATION PRODUCT(S)
English2430NwodoEnglish NJ NnadiEnglish COEnglish OkoyeEnglish FBCEnglishArtesunate contains lactone ring known to degrade in alkaline medium; a reaction that could serve as a novel, simple, precise and rapid spectroscopic tool for quantitative determination in bulk and dosage forms. The degradation of artesunate in nucleophilic agents was investigated. Absorption spectra of artesunate in ethanol, Britton-Robinson buffer, and ammonium buffer (pH 5-12) were obtained. The effects of time and temperature on the stability of the degradation products were also evaluated. Each change in pH caused the spectrum of artesunate to change significantly with time up to pH 12, where a prominent peak at 225 nm was observed. Artesunate follows linearity in the concentration range of 2-20 μg/ml with correlation coefficient of 0.999 in the proposed method. At constant temperature, therefore, the degradation products of artesunate at pH 12 could be harnessed for qualitative and quantitative determination of artesunate in dosage forms and biological fluids. The method was validated for specificity, precision and accuracy.
EnglishAbsorption maxima, pH, Buffer solution, ValidationINTRODUCTION
Malaria poses a huge economic burden on countries where the disease is endemic. According to the World Health Organization (WHO), malaria is a significant public health problem in more than 90 countries inhabited by some 2400 million people and causes an estimated 300 million acute illness each year with at least a million deaths1 . More than 90 % of these occur in Sub-Saharan Africa, and it is estimated that the disease kills an African child every 30 seconds2 . Spread and emergence of old and new drug-resistant parasites is threatening malaria control programmes in most malaria-endemic regions3 . The high morbidity of malaria necessitated the use of many antimalaria drugs in therapy, at least to control or eradicate the disease4 . One of such drugs is artesunate, a qinghaosu-based compound derived from the herb artemisia. It is a fine white crystalline powder, slightly soluble in water and freely soluble in ethanol.
Due to the extensive use of artesunate alone or in combination as artemisinin-based combination therapy ( ACT) in malaria treatment, various assay methods have been developed5,6,7. Unfortunately, these methods are costly, time consuming and non-environmental friendly. The International Conference on Harmonisation (ICH) guidelines for development of assay methods for drugs require the consideration of the regulatory status of the developed methods of assay, review of literature on stability-indicating assay8 , assessment of the extent to which the developed method meet current regulatory requirement, techniques employed in development of assay methods and development of validated stability-indicating assay methods (SIAMs) that are likely to meet regulatory requirement9 . Studies of this nature provides a finger-print for spectroscopic detection of artesunate indirectly by understanding the reaction of the quinghaosu backbone of artesunate. Quinghaosu is unstable in alkaline medium and may decompose at room temperature to form many degradation products such as octahydro-indene10. The cleavages of the lactone ring and/or epoxide linkage and succinate moiety of artesunate can be harnessed spectrophotometrically to identify the drug qualitatively and quantitatively in non-interfering matrices such as dosage forms and biological fluids.
MATERIALS AND METHODS
The major materials used for the study are Artesunate (Swiss Pharma., Nigeria), Ultraviolet/Visible Spectrophotometer (Model UNICO 2100, China) and pH meter (Eutech, Japan). All reagents for this study were used as received without further purification.
(a) Preparation of Buffer Solutions
0.4 M each of orthophosphoric acid, boric acid, acetic acid, sodium hydroxide, ammonium hydroxide, ammonium chloride and 2.0 M ammonium hydroxide were prepared. BrittonRobinson (BR) buffer (pH 5-8) and ammonium buffer (pH 8-12) were prepared as described11 .
(b) Preparation of Standard Solution of Artesunate in Ethanol
A 100 µg/ml artesunate solution was prepared by accurately weighing and dissolving 100 mg of pure artesunate powder in ethanol. The solution was made up to 1 litre with ethanol. From the stock solution, 10 µg/ml solution was prepared by serial dilution with ethanol and scanned in UV spectrophotometer against ethanol blank.
(c) Method Development
(i) Degradation Analysis of Artesunate
A 10 µg/ml solution of artesunate was prepared from the standard solution using the BR buffer at pH 5 by serial dilution. The absorption spectrum was obtained against ethanol-BR solution blank. The procedure was repeated using BR buffer at pH 6-8 and ammonium buffer 8-12. The spectra of the drug were shown in Figures 4 and 5.
(ii) Effect of Time and Temperature on the Stability of Degradants.
A 10 µg/ml solution of artesunate was prepared from the stock solution using ammonium buffer (pH 8) and the absorbances were taken after 30, 60, 90, 120 and 150 minutes of standing. The absorbances of fresh solution were taken after heating to 50, 60, 70, 80 and 90 ºC at ?max specific for each pH. These procedures were repeated for ammonium buffer (pH 9-12). Plots of the absorbance against time and temperature were shown in Figures 2 and 3 respectively.
(iii) Calibration Curve of Artesunate at Degradation Conditions.
Aliqouts of stock solution were further diluted with ammonium buffer (pH 12) to get working solution of 2, 4, 8, 12, 16 and 20 µg/ml. Subsequently, the absorbances were taken at ?max of 225 nm against ethanol-buffer blank. A calibration curve of the absorbance against concentration was plotted as shown in Figure 6.
(iv) Assay Procedure for Formulations
An amount of finely ground tablet powder equivalent to 50 mg of artesunate was accurately weighed into a 100 ml calibrated flask, 50 ml of ethanol added and shaken for 10 minutes. Then, the volume was made up to the mark with ammonium buffer (pH 12), mixed well, and filtered using a Whatman No. 42 filter paper. First 10 ml portion of the filtrate was discarded and a suitable aliquot of the subsequent portion 500 μg/ml was diluted appropriately to get concentrations for analysis by the method. The results are shown in Table 3.
(d) Method Validation
(i) Precision
The precision of the developed method was determined by measuring the absorbances of six separate samples containing known amount of drug prepared under degradation condition. The method was validated by calculating the slope, intercept, correlation coefficient and optical characteristics (ICH guidelines 1995) as shown in Table 1.
(ii) Recovery Studies
Recovery studies were carried by adding 80, 100 and 120 % of pure artesunate to different samples of tablet powder containing equivalent of 50 mg of artesunate12. From the amount of drug found, percentage recovery was calculated. The percentage recovery and the % RSD values are shown in Table 2.
DISCUSSION
The proposed method is based on the degradation of the lactone ring of the artesunate in alkaline medium and the reaction is followed spectrophotometrically for quantization purposes13. The UV absorption spectra of artesunate in ethanol and buffer solution are illustrated in Figures 4 and 5 for buffers of pH 5-8 and pH 8-12 respectively. The peak at 232 nm obtained in ethanol could be attributed to succinate moiety of artesunate while the peak at 272 nm could be due to lactone ring11. The minor changes in the maximum wavelength of absorption observed for pH 5-7 suggest the introduction of functional groups in the course of the degradation. The bathochromic shift of peak at 272 nm to 280 at pH 5 may be attributed to the cleavage of endoperoxide linkage of artesunate leading to addition of C=C bond5 . The appearance of exactly same spectra for pH 8a and 8b shows that buffer system has no interference in the degradation of artesunate. It was observed that different degradation products were formed in all the buffer solution used but only the products formed at pH 12 was stable on standing. Assuming that absorbance is proportional to concentration, as stated in BeerLambert’s law, the the decrease in absorbance as temperature increases indicates a decrease in the concentration of the product. This signifies that the product is not very stable to temperature. At constant temperature, therefore, the product can be said to be stable as the variation in the absorbance was not pronounced as shown in Figures 2 and 3. The reliability, accuracy and validity of the proposed method were ascertained by carrying out recovery studies. The recovery of pure drug added was quantitative and revealed that co-formulated substances (excipients) did not interfere in the determination as shown in Table 2. This further stresses the selectivity of the method for analysis of artesunate. In order to check the validity of the proposed method, analysis of artesunate in some commercial formulations was carried out. Table 3 shows the results of studies in which there is close agreement between the result obtained by the proposed method and label claims indicating high accuracy and precision. The proposed method is simple, precise and useful in routine determination of artesunate in pharmaceutical dosage forms and biological fluids. Research is currently in progress to isolate, characterize and elucidate the structures of the degradation product(s) of artesunate for more accurate on-the-spot detection of the drug.
CONCLUSION
A method for indirect determination of artesunate in bulk and dosage forms has been developed and validated in the concentration range of 2-20 µg/ml (R2 = 0.999) with average percentage recovery of 99.93 from tablets. The developed method is selective, novel, precise, linear, simple accurate and suitable for routine determination of artesunate without interference from dosage matrices.
ACKNOWLEDGEMENT
The authors are grateful to Chidera Charles-Nnadi for her technical support and to the Stephen Oluwole Awokoya Foundation for Science Education, Lagos Nigeria for providing financial assistance. 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=1992http://ijcrr.com/article_html.php?did=19921. World Health Organization Global Report on Antimalaria Drug Efficacy and Drug Resistance. WHO Geneva, 2010.
2. Range HP, Dale MM, Ritter JM, Flower RJ. Range and Dale’s Pharmacology. Elsevier 6(1) China. 2007; 467-474.
3. Kamau E, Alemayehu S, Feghali KC, Tolbert LS, Ogutu B, Ockenhouse CF. Development of a TaqMan Allelic Discrimination Assay for Detection of single nucleotides polymorphisms associated with anti-malarial drug resistance. Malaria Journal. 2012; 11:23 doo, 10.1186/1475-2875-11-23. http//www.malariajournal.com/content/11/1/2 3.
4. Boulangier D, Dieng Y, Cisse B. Antischistosomal Efficacy of artesunate combination therapies administered as curative treatment for malaria attacks. Tropical Med. Journal. 2007; 10(102): 198- 265.
5. Thekke VS, Badiadka N. A Simple and Rapid Spectrophotometric Method for the Determination of Artesunate in Pharmaceuticals. Eurasian J. Anal. Chem. 2009; 4(1): 119-126.
6. Esimone CO, Omeje EO, Okoye FBC, Obonga WO, Onah BU. Evidence for the spectroscopic determination of Artesunate in dosage form. J Vector Borne Dis 45, pp. 2008; 281–286.
7. Olajire AA, Olakunle S.I, Oluwafunmibi PD, Olutayo SO. Derivatization of artemisinin derivatives using 4-carboxyl -2, 6-dinitro benzenediazonium (CDNBD) ion Acta Pharmaceutica Sciencia. 2010; 52: 269-280.
8. . Monika B, Saranjit S. Development of validated stability-indicating assay methods: critical review. Journal of Pharmaceutical and Biomedical Analysis 2002; 28:1011–1040.
9. Stability Testing Of New Drug Substances And Products Q2 (R1) Nov. 2005; ICH Harmonized Tripartite Guideline.
10. Liu JM, Ni MY, Fan YF, Tu YY, Wu ZH, Wu YL, Chou WS. Structure and Reactions of Artemisinin. Huaxue Xuebao, 1979; 37(2): 129–141.
11. Viviane MI, Leticia NCR, Claudia CIG, Maria VBZ. Spectrophotometric determination of Cefaclor in Pharmaceutical Preparation. Quimica Nova 1998; 22(2): 801-807.
12. Jeswani RM, Sinha PK, Topagi KS, Damle MC. A Validated Stability Indicating HPTLC Method for Determination of Cephalexin in Bulk and Pharmaceutical Formulation International Journal of PharmTech Research. 2009; 1(3): 527-536.
13. Basavaiah K, Anil Kumar UR. Simple Spectrophotometric Methods for the Determination of Zidovudine in Pharmaceuticals Using Chloramine-T, Methylene Blue and Rhodamine-B as Reagents. E-Journal of Chemistry. 2006; 3(12):173-181.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesPARTIAL PURIFICATION AND CHARACTERIZATION OF DETERGENT TOLERANT CELLULASE FROM BACILLUS SP. KP8
English3137Dhairyawant KushwahaEnglish Harsh Dev KumarEnglish Gireesh Babu KEnglishIn this study cellulase producing Bacillus sp. KP8 was isolated from soil. The crude cellulase was found to possess 0.012μmoles/ml/min activity. The purification using Sephadex G-100 and DEAE-Cellulose lead to ~189 fold purity with 9.09% recovery. The enzyme was found to be optimally active at pH 5.0, 45ºC. The partially purified fraction displayed two proteins of ~45 kD. The kinetic studies revealed a Km of 2.85, while the Vmax was found to be 5.9. The non-metallo cellulase was detergent stable, possessing serine at its active site, sensitive to the presence of Mg+2, Mn+2, Hg+2, Cu+2, H2O2 and NaCl.
EnglishCellulase, Bacillus sp. KP8, Sephadex G-100, DEAE-Cellulose, SDS-PAGEINTRODUCTION
Today, the enzymes are commonly used in many industrial applications and the demand for more stable, highly active and specific enzymes is growing rapidly. Cellulolytic enzymes play an important role in natural biodegradation processes of plant lignocellulose [1] by cellulolytic fungi, bacteria, actinomycetes and protozoa [2]. High cost of cellulase is mainly due to the substrates used in production and also the slow growth rate of fungi [3]. Bacteria, which has high growth rate as compared to fungi has good potential to be used in cellulase production. However, the application of bacteria in producing cellulase is not widely used. Bacterial cellulase usually lacks one of the three cellulase activities. However, cellulases produced by bacteria are often more effective catalysts. They may also be less inhibited by the presence of material that has already been hydrolyzed (feedback inhibition). Cellulose is the most abundant bio-polymer on earth; an estimated 7.5×1010 tons is annually synthesized through photosynthetic processes [4]. Found primarily in plant cell walls, cellulose is embedded in a heteromatrix composed of xylan, other hemi-celluloses and lignin. Specifically, cellulose is a linear, insoluble biopolymer composed of repeating β–Dglucopyranose residues linked by β-1, 4 glycoside bonds. In contrast to other glucan polymers, such as starch, the repeating unit of cellulose is not glucose, but cellobiose, a disaccharide. Cellulose exhibits a high degree of polymerization: the individual glucan chains, or cellodextrins, can reach lengths of greater than 25,000 glucose residues. The present study reports the purification and biochemical characterization of cellulase from Bacillus sp. KP8.
MATERIAL AND METHODS
Bacteria The Bacillus sp. KP8 isolated from the soil was used in the present study.
Production of cellulase
The Bacillus sp. KP8 was grown in 250ml nutrient broth with 0.5% CMC for 24h. The cells were harvested by centrifuging at 10,000 rpm for 3min and supernatant was used as the crude enzyme sample for cellulase assay and purification.
Cellulase Assay [5]
The 2ml reaction mixture of the enzyme contained 1ml of 0.1% CMC, 0.5ml of acetate buffer (25 mM, pH 5.0) and 0.5 ml of enzyme extract. The tubes were incubated at 450C for 10 min and added with 1ml of DNS. The tubes were further boiled for 10min, cooled and the absorbance was measured at 540 nm against the blank, added with DNS before the addition of enzyme. The concentration of glucose released was estimated by standard glucose curve. The activity was calculated according to the following formula;
Preparation of standard glucose curve
The different concentrations of glucose (0-100μg) were added into different tubes, by pipetting from the stock solution of glucose (1mg/10 ml). The mixture volume was made up to 2 ml by adding water followed by the addition of 1ml of DNS. The tubes were boiled for 10 min, cooled and the absorbance was read at 540 nm, against the blank (Fig 1).
Estimation of Protein [6]
The protein content of samples at every stage of purification was determined by the method of Bradford (1976). Bradford reagent was prepared accordingly. 20 mg CBB G-250 dissolved in 10 ml of 95% ethanol and 20 ml of orthophosphoric acid and made up to 200ml with distilled water. For the protein estimation, to 5ml of Bradford reagent 100μl of protein sample was added, mixed well and incubated for 5min at room temperature. The color developed was checked for its absorbance at 595nm against Bradford reagent as blank.
Standard BSA Curve
The standard Bovine Serum Albumin curve was prepared by taking the absorbance of different concentrations of BSA (0-100μg) mixed with 5 ml of Bradford reagent, at 540 nm. The graph was plotted taking concentrations of BSA on X axis and absorbance on Y axis (Fig 2).
Purification of Cellulase Precipitation of proteins by increasing the ionic strength
The crude protein supernatant was precipitated with ammonium sulphate (till 75% saturation) at 4 0C with constant stirring. The proteins were retrieved by centrifuging at 10,000 rpm for 10 min at 40C. The pellets of proteins were dissolved with minimum volume of 50mM Acetate buffer (pH 5.0).
Dialysis
The protein sample was loaded into the preactivated dialysis membrane (Himedia, cut off range 11 kD). The dialysis membrane was then hung into the 500ml of acetate buffer (5mM, pH 5.0). The dialysis was carried out for 8 h with 2 changes in buffer, at 40C. Gel filtration chromatography The Sephadex G 100 (Sigma-Aldrich) column (50X1cm) was run with 10 times the volume of acetate buffer (50 mM, pH 5.0). The 3 ml dialyzed enzyme sample was loaded and 25 fractions (2 ml) were collected. The fractions were analyzed for protein and enzyme activity. Ion-exchange chromatography The pre-activated DEAE-Cellulose (Bangalore Genei, Bengaluru) column (25X2 cm) was washed with 10 volumes of acetate buffer (50 mM, pH 5.0) and loaded with the pooled active fractions of gel filtration column. The cellulase was eluted by gradient buffer concentration (50-500 mM). The eluent was collected in 25 fractions of 2 ml each. The fractions were analyzed for protein and cellulase activity.
Characterization of cellulase
Polyacrylamide Gel Electrophoresis (PAGE)
The partially purified cellulase was further analyzed by SDS-PAGE [7]. The 10% gel was loaded with sample and molecular weight marker (medium range, Bangalore Genei, Bengaluru) and electrophoressed with a constant power supply of 100 V using electrophoresis buffer (Tris- 3g, Glycine- 14.3g, SDS- 2.0g for 1 liter). The gels were stained using silver staining technique. The enzyme found to be ~45 kD.
Biochemical characterization
Effect of pH on Cellulase Activity
The enzyme was assayed in presence of different pH buffers (50 mM), Citrate buffer (pH 4.0), acetate buffer (5.0 and 6.0), phosphate buffer (7.0) and tris buffer (8.0) were used.
Effect of Temperature on Cellulase
Activity Effect of temperature on cellulase activity was determined by incubating the reaction mixture at different temperature ranging from 37, 45 and 550C.
Kinetic studies of cellulase:
To study the kinetic properties of enzyme, the enzyme was assayed with different concentrations of CMC. The total volume of buffer and substrate in the test tube was made up to 1 ml. The Line weaver Burk plot was used to find the Vmax and Km.
Effect of Inhibitors and Metal ions and Inhibitors
The effect of various cellulase inhibitors and metal ions were analyzed. The enzyme was incubated for 10 min with different metals (20mM), inhibitors (2mM), H2O2 (30%), 1% SDS (Sodium Dodecyl Sulphate) 1M NaCl (Sodium Chloride) and assayed as mentioned above.
RESULTS
Cellulase activity The Bacillus sp. KP8 used in the present study could produce 0.012 µmoles/ml/min cellulase. Purification of cellulase By various steps of purification, the cellulase was purified by 188.88 folds with 9.09% yield (Table1). The SDS-PAGE analysis revealed presence of two bands ~45 kD (Fig.3).
Biochemical characterization Effect of pH on cellulase
The cellulase was assayed at various pH and found to be more active at pH 5.0 (Fig 4). Effect of temperature By incubating the reaction mixture at different temperatures, the highest activity was noted at 450C (Fig 5).
Kinetic studies
The LB plot (Fig. 6) drawn by the collected data indicated that the Km value as 2.85, while the Vmax was found to be 5.9.
Effect of metal ions and inhibitors on cellulase activity
Various metal ions, inhibitors and additives were tested for their effect on cellulase activity (Table2). The enzyme was completely inhibited by Mg+2, PMSF and EDTA. No additive could induce the activity.
DISCUSSION
This work demonstrated that Bacillus sp. KP8 isolate as good producers of cellulose. Cellulolytic species are found within the phyla Thermotogae, Proteobacteria, Actinobacteria, spirochaetes, Firmicutes, Fibrobacteres and Bacteroids. Of these, approximately 80% of the isolated cellulolytic bacteria are found within phyla Firmicutes and Actinobacteria [8]. The majority of the gram-positive cellulolytic bacteria is found within Firmicutes and belongs to the class Clostridia and the genus Clostridium. Although first described within Clostridium thermocellum, numerous other clostridial species have been shown to utilize a cellulosome as their cellulose degrading strategy. These bacteria are ubiquitous in anaerobic soil environments and include C. cellulolyticum, C. acetobutylicum, C. cellobioparum and C. papyrosolvens [9]. The cellulase was purified by 188.88 folds with a recovery of 9.09%. A major loss in the recovery was noticed during the gel filtration process. The SDS-PAGE indicated the presence of two proteins of ~45 kD, which have to confirmed for isoforms. The cellulase was optimally active at acidic conditions requiring moderately high temperature (450C). The enzyme was found to possess serine at its active site as it lost its activity completely in presence of PMSF. The enzyme was also found to be inhibited by Mg+2 and EDTA, indicating the non-metallo nature. The cellulase was detergent stable as it could retain 98% activity in presence of SDS, but not bleach stable and salt tolerant as it lost activity in presence of H2O2 and NaCl, respectively. The present work reports the bacterial cellulase, which has to be studied further for the suitability and applications in industry.
Englishhttp://ijcrr.com/abstract.php?article_id=1993http://ijcrr.com/article_html.php?did=19931. Lynd, L.R.; van Zyl, W.H.; McBride, L.E.; Laser, M. Consolidated bioprocessing of cellulosic biomass: an update. Curr Opinion in Biotechnology 2005, 16(5), 577-583.
2. Olsson L., Hahn-Hagerdahl B. 1997. Fermentation of lingocellulose hydrolisates for ethanol production. Enzyme Microb. Technol. Vol. 18. P. 312–331.
3. Lynd, L.R.; Weimer, P.J.; van Zyl, W.H.; Pretorius, I.S. Microbial cellulose utilization: Fundamentals and biotechnology. Microbiology and Molecular Biology Reviews 2002, 66, 506- 577.
4. Monserrate, E.; Leschine, S.B.; Canale-Parola, E. Clostridium hungateisp. Nov., a mesophillic, N2-fixing cellulolytic bacterium isolated from soil. International Journal of Systematics, Evolution and Microbiology 2001, 51, 123-132.
5. Eriksson, K.E. and Petterson. B., 1972. In Biodeterioration of materials. J. Biotech. 2: 116.
6. Bradford, M.M. (1976) A Rapid and sensitive method for quantification of microgram quantities of protein utilizing the principles of protein dye-binding. Ann. Biochemistry. 32:248-253.
7. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4, Nature 227:680-685.
8. Bergquist, P.L.; Gibbs, M.D.; Morris, D.D.; Te’O, V.S.; Saul, D.J. Morgan, H.W., Molecular diversity of thermophillic cellulolytic and hemicellulolytic bacteria. FEMS Microbiology Ecology 1999, 28, 99-110
9. Desvaux, M. Clostridium cellulolyticum: model organism of mesophillic cellulolytic clostridia. FEMS Microbiology Reviews 2005, 29, 741- 764.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesNANOPROCESS FOR WATER PURIFICATION
English3853V. MadhaviEnglish P.Jeevan JyothiEnglish A.Vijay Bhaskar ReddyEnglish K.Gangadhara Reddy G.MadhaviEnglishNanotechnology is an emerging science that has shown potential applications in solving current problems involving water quality. Nanomaterials are highly reactive, have a large surface area compared to their mass and can easily capture other particles. Hence nanotechnology is also used as an environmental technology to protect the environment through pollution prevention treatment and clean up. Additionally, anotechnology-derived products can reduce both chemical and biological impurities in water treatment. This article summarizes in the area of nanomaterial synthesis and the origin of their reactivity at the nanoscale in environmental clean up like water purification. In addition, we discuss the limitations associated with the potential applications of nanomaterials for water purification.
Englishchemical and biological impurities, environmental technology, nanomaterials, potential applications, water qualityINTRODUCTION
Environmental pollution is a serious day-to-day problem faced by the developing and developed nations in the world. Due to increased industrialization and urbanization a vast majority of water quality problems are caused by contamination, overexploitation or combination of the two, soil and water quality is slowly but declining everywhere. The common pollutants include toxic compounds like chlorinated and nonchlorinated aliphatic and aromatic compounds, dyes, detergents and surfactants, agro wastes like insecticides, pesticides and herbicides, disinfection byproducts, volatile organic compounds, plastics, inorganic compounds like heavy metals, noxious gases like NOx, SOx, CO, NH3and pathogens like bacteria, fungi and viruses. Clean water which is free of toxic chemicals and pathogens is essential for the very existence of life. Clean water is also a critical feedstock in a variety of key industries including electronics, pharmaceuticals and food. The world is facing formidable challenges in meeting the rising demands of clean water. In order to secure water resources water reuse is becoming more viable. One of the concerns with water reuse is the contamination with chemicals, bacteria and other pollutants. Various environmental technologies have been employed to remove the pollutants in water. Conventional water treatment plants are constructed based on three important assumptions. Firstly, the influent source waters entering the treatment plant is comprised of only naturally occurring chemical and biological contaminants. Secondly, they appear in the source waters mainly due to surface water runoffs, localized conditions existing in the source waters and cross contamination resulting from discharge of untreated sewage. Thirdly, the contaminants present in source water can be completely removed via a simple treatment following a sequence of steps inclusive of coagulationflocculation, filtration and disinfection.1 However, to date neither type of existing conventional treatment is universally applicable or highly effective. This suggests that there is a need for technological advancement in water treatment to benefit people in many countries. In recent years, nanoscience and nanotechnology has introduced a new dimension to scientific discipline and technology sectors due to its ability to exhibit super functional properties of materials at mono-dimensions. There is a great potential to use this technology to clean up the contaminated sites and protect the environment from the pollution. This eco-friendly technology is considered to be an effective alternative to the current practices of site remediation. Nanoremediation methods involve application of reactive materials for the detoxification and transformation of pollutants. Nanomaterials will enable new means of reducing the production of wastes, using resources more sparingly, cleaningup chemical and bacterial contamination, proving potable water and improving the efficiency of energy production and use. Several nanomaterials include magnetic nanoparticles, heterogeneous nanophotocatalysts and polymeric nanoparticles. Due to their strong magnetic properties, magnetic nanomaterials act not only as an adsorbent to remove target compounds from the contaminated water, but also as a magnetic element to attract and retain the nanoparticles, which can be removed from solutions. This magnetic separation, which may replace centrifuge separation technologies, has less complicated technical requirements and low generation cost, thus making this adsorption treatment economically attractive for individual users.2 Many different nanomaterials have been evaluated for use in nanoremediation. They include nanoscale zeolites, metal oxides, carbon nanotubes, noble metals and titanium dioxide. This review focuses on various research works regarding the use of nanotechnology for environmental clean-up, particularly on their application in remediation of water.
Iron based technologies
Iron is the fourth most abundant element in the earth’s crust, and reactions involving iron play a major role in the environmental cycling of contaminants. Iron exists in the environment in two valence states- water soluble Fe (II) and highly water insoluble Fe (III). Zero valent Iron (Fe (0)) is also found under some specific and environmental conditions.
Various mineral forms of iron
In the environment, iron plays an important role in contaminant mobility, sorption and breakdown due to its role as an electron donor, and, in its various mineral forms, as a precipitant/ sorbent surface. Since early 1990s, the iron corrosion has been put into productive use in the treatment of hazardous and toxic chemicals. The use of iron-based technologies in contaminated water remediation is a rapidly developing field, with a range of techniques proposed which make use of iron as a reductant, precipitant and sorbent. The applications of iron-based technologies in contaminated water remediation can be broadly divided into two, based on the chemistry involved in the remediation process. Technologies which use iron as 1) a sorbent, precipitant or immobilizing agent 2)as an electron donor to convert contaminants into a less toxic form. The ability of iron, both of its zero valent form and as its Fe2+, to reduce redox sensitive elements like Cr, Tc and to dechlorinate various organic contaminants has been successfully used. Reactions for Cr reduction and immobilization include:
Fe2+ + CrO4 - + 4H2O (Fex(Cr1-x) (OH)3 + 5OHin which the toxic Cr (VI) can be reduced to Cr (III) form, which readily precipitates as Cr(OH)3 or as a solid solution (Fex(Cr1-x) (OH)3 (Puls et al.,1999). Elemental iron slowly oxidizes to ferrous iron and releases two electrons. Contaminants such as polychlorobiphenyls, or PCBs, and chlorinated benzenes can accept the electrons and be reduced to hydrocarbon compounds. For example, tetrachloroethene (C2Cl4) can be reduced to ethene in accordance with the following stoichiometry: 4Fe0 + C2Cl4 + 4 H+ C2H4 + 4Fe2+ + 4ClFerrous sulphate and Ferrous Ammonium Sulphate can also be used in treatment of contaminated water. Ferrous Sulphate is a traditional reducing agent for the treatment of metal industry process effluents. Ferrous Ammonium Sulphate has the advantage over ferrous sulphate of reacting relatively rapidly over neutral to alkaline pHs, avoiding the need for acidification.3The ability of iron to act as an electron donor or reducing agent is utilized in Fenton treatment technologies, where Fe2+ or Fe0 can be used to reduce Hydrogen peroxide and generate the highly reactive OH radical. Fe0 + H2O2 →Fe 2+ +OHFe2+ can then react with H2O2 in traditional Fenton’s oxidation reactions Fe2+ +H2O2 Fe3+ + OH + OH ? - In the presence of H2O2, Fe0 is transformed into Fe2+ Fenton techniques show considerable efficiency in the remediation of pesticides, fuels, explosives etc.4 observed a 90% reduction of chlorinated contamination in water. Fenton treatment can be applied for the removal of organic and inorganic contaminants in situ. Of many iron-based materials, nZVI (nano Zero Valent Iron) are generally preferred for remediation because of large surface area of nanoparticles and more number of reactive sites than micro-sized particles5 and it possess dual properties of adsorption and reduction. Solutionphase and Vapor-phase synthesis methods can be used for producing iron at nano-scale. Since vapor-phase synthesis normally yields relatively small quantities of particles, most of the synthesis approaches which are commercially available or normally used in laboratories, are solution-phase methods. Iron at nano-scale can be synthesized by solution-phase method is from Fe (II) and Fe(III) , using borohydride as a reductant: 4Fe3+ + 3BH4 - +9H2O 4Fe0 + 3H2BO3 - + 12 H+ +6H2 The excessive borohydride is typically needed to accelerate the synthesis reaction and ensure uniform growth of iron crystals. Synthesis at much lower concentrations and the use of ferrous iron has also successfully performed. Emulsification of nZVI is another novel modification of nZVI for active in situ remediation. This approach is adapted for emulsified oil flushing for DNAPLs (dense non-aqueous phase liquids), such as trichloroethane (TCE), which are hydrophobic. The emulsion is miscible with the contaminant, allowing an increased contact between TCE DNAPL and the ZVI present within the oilemulsion droplet6 . The use of iron as a permeable reactive barrier (PRB) has been the subject of considerable research and development since 1990s7 . PRB is an engineered zone of reactive material, extending below the water table, designed to treat contaminated water. Contaminants passing through PRBs are either degraded or retained in the reactive barrier material. ZVI has been used particularly as a reactive media in a number of field-scale PRB systems particularly those designed to remediate chlorinated organic compounds, metal and radionuclide contaminants successfully8 . PRBs containing nZVI may remove organic chlorinated compounds by reductive dechlorination9 , whereas metals, metalloids and radio nuclides can be removed by reductive precipitation, surface adsorption or complexation or co-precipitation with the Fe oxyhydroxides that form on the nZVI surfaces10.Iron with other metals (bimetals) was used extensively for the remediation of contaminated water. Bimetallic nanoparticles consist of elemental iron in conjugation with a metal catalyst, such as platinum (Pt), gold (Au), nickel (Ni) and palladium (Pd)11 . The combination of metals to form a nanoparticle increases the kinetics of redox reaction, therefore catalyzing the reaction. The most commonly used and commercially available bimetallic nanoparticles (BNPs) are the palladium and iron BNPs (Pd/Fe).The surface area normalized rate constant of BNPs of Pd/Fe was two orders of magnitude higher than that of nZVI. Pd/Fe BNPs are generally used in the removal of trichloroethane.
Carbon nanotubes
In recent years, nanotechnology has introduced different types of nanomaterials to the water industry and has produced some promising outcomes. With emergence of nanoscience and technology in the last decade, research has been initiated to exploit the unusual and unique properties of carbon nanotubes (CNTs). CNTs are very thin; hallow cylinders made of carbon atoms. They are about 10,000 times thinner than human hair. CNTs, a new form of carbon, are attracting great research interest due to their exceptional adsorption, mechanical properties, unique electrical properties and high chemical and thermal stability mainly because of their extremely small sizes, uniform pore distribution and large specific surface area12. CNTs are nanomaterials that are rolled into a tube and are classified as single-walled carbon nanotubes (SWNTs), a single pipe with a diameter from 1 to 5 nm and multiwalled carbon nanotubes (MWNTs) with several nested tubes, at lengths varying from 100 nm upto several tens of micrometers. The SWNTs bundles have their adsorption sites inside the tubes, the interstitial triangular channels between the tubes, or the grooves formed at the contact between adjacent tubes on the outside of the bundle. For MWNTs, adsorption can occur in the aggregated pores, inside the tube, or in the external walls. Since their discovery, CNTs have attracted great attention due to their unique properties. CNTs can be synthesized by chemical vapor deposition (CVD) process. The characteristics of CNTs synthesized by this process depend upon the type of catalyst and carrier gas used13.A novel silica template-mediated approach for synthesis of nanoporous carbon using CVD was proposed by Ryoo et al14.The resulting high surface area materials with uniform pores were suitable for wide range of applications such as adsorbents, catalytic supports etc. The dye adsorbing capacity of such nanoporous carbon was found to be 10 times higher than that of commercial activated carbon.
Classification of CNTs
The hexagonal arrays of carbon atoms in the graphite sheets of CNTs surface have a strong interaction with other molecules or atoms, which make CNTs a promising adsorbent material substituted for activated carbon in many ways15 . These are utilized for the removal of heavy metals, metalloids, organic and biological impurities. Nanotube surfaces are chemically modified to enhance the adsorption of contaminants. The functionalization with various groups such as – OH, -COOH, -NH2 etc. and introduction of nanopores in activated carbon by chemical oxidation (by HNO3, KMnO4, H2O2, NaOCl, H2SO4, KOH, and NaOH etc.) and heat treatment was another approach for synthesis of nanoporous carbon which increases their water solubility and biocompatibility. Heavy metal adsorption on CNTs has been shown to depend on surface functional groups, specific surface area, and solution components. The most important factor is the surface functional group, which generates oxidized acids. It is well known that oxidation treatment by nitric acid causes an increase in cation exchange capacity. Due to their porous structure, CNTs are found to have much higher adsorption than that of carbon black with the same surface area of CNTs. The researchers assert that CNTs are effective adsorbents for environmental applications when compared to other adsorbents. The unique properties of CNTs like high chemical and thermal stability were utilized for the treatment of natural organic matter (NOM) which produces carcinogenic agents, thus maintain high water quality. CNTs have significantly higher dioxin removal efficiency than that of activated carbons. CNTs are good fluoride adsorbents and their fluoride removal capacity is superior to that of an activated carbon filter. Adsorption of 2,3- dichlorophenol was found to be dependent on the mass of MWCNTs , concentration, solution pH and adsorption temperature. The amino functionalized MWCNTs can be used to produce filtration membranes for the removal of heavy metals from industrial waters. Li et al.16 found that the metal ion sorption capacities of the MWNTs were 3-4 times larger than those of powdered activated carbon and granular activated carbon, two commonly used sorbents in water purification. Peng et al.17 have recently developed Cerium-oxide supported on carbon nanotubes which are effective sorbents for As (V). Li et al.18 reported that MWNTs were better sorbents of volatile organic compounds (VOCs) than carbon black in aqueous solutions. Fuget su et al.19 has prepared cross-linked alginate vesicles encapsulated inside MWCNTs which have high sorption capacity for water soluble dyes. CNTs have also unique strength in adsorption of biological contaminants because of their structural and functional properties. Anti-microbial effect of CNTs is due to their fibrous shape20. The groove edges of CNT bundles and the external surface area of outermost nanotubes are potential adsorption sites and provide large pore spaces that will be fully utilized by micro-organisms. Thus with respect to adsorption of biological contaminants on CNTs, accessible external surface area and presence of aggregated pores with volumes greater than mesopore are considered important. The microbial cytotoxic property of CNTs has a partial influence on concentration of bacteria. Thin fibers of CNTs impinge bacterial cell surface, disrupt the intracellular metabolic pathways and subsequently, the internal contents are released due to the cell rupture caused by oxidative stress after impingement21.The size and length of the tubes, dispersivity, amorphous nature and number of layers (single or multi walled) are identified to influence the cytotoxic properties of CNTs22. Bacterial adsorption on CNTs is characterized by having three unique features: 1) microbial adsorption capacities on CNTs are higher than any other commercially available adsorbent media. 2) CNTs express selective adsorption of bacteria, a feature which is generally not seen in other adsorbents. 3) Adsorption kinetics of bacteria on CNTs is almost instantaneous. The pristine modified CNTs prohibit the growth of pathogens on their surface and might probably contribute to self-cleaning efficiency of the CNT absorption filters. Pristine CNTs exhibited antimicrobial characteristics over wide range of micro-organisms including a) bacteria e.g. Micrococcus lysodeikticus, Streptococcus mutans, E.coli, Salmonella and bacteria endospores 22-24. b) Protozoa species e.g., Tetrahymena pyriformis, 25 c) viruses, e.g., MS2 bacteriophage 26-27.
Silver nanoparticles
Water is the common breeding ground for many micro-organisms. The greater water-borne threat to human health is bacterial contamination of drinking water sources leading to outbreak of diseases. The removal or inactivation of microorganisms is the last step in the treatment of waste water. Many chemical and physical agents such as chlorine and its derivatives, ozone, AgNO3, UV light are commonly used for disinfection of water28 which reacts with various constituents in natural water to form disinfectant byproducts (DBPs), many of which are carcinogens. DBPs will be formed when chemical oxidants are used in water treatment. Furthermore, the resistance of some pathogens, to conventional chemical disinfectants requires high disinfectant dosage, leading to higher DBP formation. The rapid growth of nanotechnology has opened significant interest in the environmental applications. The use of metal nanoparticles for disinfection is expected to play a crucial role in water purification because of their high reactivity due to the large surface area to volume ratio. These nanoparticle scan either directly by interacting with the microbial cells, e.g. interrupting trans membrane electron transfer, disrupting/penetrating the cell envelope, or oxidizing cell components, or producing secondary products(e.g. reactive oxygen species or heavy metal ions) that cause damage. The antibacterial properties of silver compounds and silver ions have been historically recognized and applied in a wide range of applications from disinfecting medical devices and home appliances to water treatment. AgNPs exhibit additional antibacterial capabilities which are not exerted by bulk or ionic silver. Current AgNPs used for disinfection in different forms include: metallic silver nanomaterials29 , silver-impregnated zeolite powders and activated carbon materials30, dendrimer-silver complexes31 , polymer-silver nanoparticle composites32, silvertitanium dioxide composite powders33 , AgNPs coated onto polymers like polyurethanes34 etc. The most common method of preparation of silver nanoparticles is chemical reduction of a silver salt dissolved in water with a reducing agent such as NaBH4, citrate, glucose, hydrazine and ascorbate etc 35-36 . Since the use of chemical reducing agents for silver nanoparticle synthesis is often considered toxic, the green synthesis methods for silver nanoparticles such as polysaccharides, polyphenols, irradiation, biological reduction are used37 . To date, several mechanisms have been postulated for the antimicrobial property of silver nanoparticles :(1) adhesion of nanoparticles to the surface altering the membrane properties: AgNPs interact with the bacterial membrane and are able to penetrate into the cell. AgNPs have been reported to degrade lipopolysaccharide molecules, accumulate inside the membrane by forming pits and cause increase in membrane permeability and cytoplasm leakage38. (2) dissolution of AgNPs releases antimicrobial Ag+ ions: The possible mechanisms for the oxidative dissolution of AgNPs have been reported by Choi et al39 and Asharani et al40 . 4Ag+ + O2 + 2H2O → 4Ag+ + 4 OH (Choi et al) 2Ag + H2O2 + 2H+ → 2Ag+ +2H2O ( Asharani et al) Ag+ has known antimicrobial properties. Ag+ interact with thiol groups in proteins, resulting in inactivation of respiratory enzymes and leading to the production of reactive oxygen species (ROS) 41 (3) generation of ROS which cause DNA damage: ROS species are natural byproducts of the metabolism of respiring organisms. Excess ROS production can lead to breakdown of mitochondrial function or cause DNA damage42 .
The factors influencing AgNP toxicity are particle size, shape, crystallinity, surface chemistry, capping agents etc. The environmental factors include pH, ionic strength, macromolecules divalent cations and presence of ligands. The decrease in particle size increases the specific surface area of AgNPs which has a higher number of atoms exposed on the surface available for redox, photochemical, biochemical reactions in addition to physico-chemical interactions with cells. AgNPS are effective biocides against 1) bacteria such as E.coli, S.aureues, B. subtillis, Klebsiella mobilis, Staphylococcus epidermis etc 43-44. 2) fungi such as A.niger, candida albicans, saccharomyces cerevisia, penicillium citrinum 45-46 3) virii such as HIV-1, Hepatitis B, Syncytial virus. 47-49 TiO2 TiO2 is the most commonly used semiconductor photocatalyst which has been applied for various photocatalytic reactions for its high efficiency, low cost, physical and chemical stability, widespread availability and non-corrosive properties. The applicability of TiO2-based heterogeneous photocatalysis has been used extensively for environmental decontamination purposes50. TiO2 nanoparticles can serve both as oxidative and reductive catalysts for organic and inorganic pollutants. It can be successfully used for treating water contaminated with dissolved metals such as Ag, Au, Hg, Cr, Pt, Cu, Ni etc.51 TiO2 nanoparticles can also completely degrade the organic pollutants into harmless inorganic substances such as CO2, H2O etc.52 under moderate conditions, and would not bring any other serious secondary pollution. According to Pirkanniemi and Sillanpaa 53, the overall heterogeneous photocatalysis can be summarized into the following five steps: 1) reactant diffusion to catalyst surface, 2) adsorption of the reactant onto the surface, 3) chemical reaction on the catalyst surface, 4) desorption of final products of the catalyst surface, 5) diffusion of final products from the catalyst surface. Heterogeneous photocatalysis process is a combination of charge transfer features, electronic structures, excited life spans and light absorption effects. The energetic and charge transfer processes involved in a photocatalytic process can be illustrated in the given figure.
When electrons are excited by the light of energy equal to or exceeding its band gap energy, they are promoted from the valence band to the conduction band, leaving positive holes in the valence band. These electrons and holes are capable of reducing and oxidizing compounds at the TiO2 surface, respectively. If electrons and holes do not recombine to produce heat, they can follow the reductive and oxidative pathways indicated by the reactions. In addition, these holes often react with water or hydroxyl ions adsorbed to TiO2 producing hydroxide radicals, which then oxidize adsorbed organics. The TiO2 occurs in rutile, anatase and brookite phases but the anatase phase of TiO2 is photocatalytic active: : TiO2 – hν →TiO2 (h+ + e) electron-hole pair formation e - +h+ → heat recombination e - +Mn+ → M(n-1) reduction h + + H2O (ads) → ?OH + H+ oxidation of adsorbed water h + + 2OH(ads) → ?OH + OH oxidation of hydroxide ions ?OH +R (ads) → ?R(ads) + H2O organic oxidation ?R(ads) (?OH, ?R(ads)) → products termination Nano-TiO2 is normally synthesized using various titania precursors such as titanium tetra-isopropoxide 54, tetra butyl titanate 55, titanium tetrachloride 56. Different starting materials can influence the morphology of the nano-TiO2 produced such as specific surface area, crystalline phase and crystallite size that plays an important role in the photocatalytic degradation of organic pollutants. Doping techniques have been applied in photocatalysis to overcome the limitations of nano-TiO2 such as a wide band gap, ineffectiveness of photocatalysis under the sunlight and thermal stability57. It is well known that small crystallite size, high percentage of the anatase phase and high specific surface area of nano-doped TiO2 increases the photocatalytic degradation efficiency. Many dopants such as metals, transition metals, metalloids, non-metals, halogens can be successfully used. Zang et al58 produced a Mn-doped TiO2 in which the band gap of the photocatalyst was narrowed due to the formation of an impurity level near the bottom of the conduction bands. Sakthivel et al59 investigated the performance of TiO2 after supplementing with platinum dopant which act as an electron trap in the formation of TiO2, decreasing its surface area. Non-metal dopants including carbon, nitrogen and sulphur are able to improve the morphology and photocatalytic performance of TiO2 60. Doping with non-metal anions broadens the band gap of TiO2 in its electronic structure and affects the red-shift in the absorption spectra of nano-doped- TiO2. Thus various photocatalytic, photochemical and the photoelectrochemical properties of TiO2 are enhanced by shifting the wavelength sensitivity of TiO2 from UV region into the visible light region61. Halogen dopants such as Iodine, Bromine and Flourine improve the morphology and the photocatalytic performance of TiO2 62. Wang et al63 successfully prepared single anatase phase of Iodine-doped TiO2 which hindered the growth of crystal particles by enhancing the energy barrier mutual diffusion between grains. Boron is known as metalloid compound that can be doped in TiO2 which inhibits the growth of crystalline TiO2 and thus increases the surface area as well as inducing the crystalline process 64 . The removal of total organic carbon from water contaminated with organic wastes is greatly enhanced by the addition of TiO2 nanoparticles in the presence of UV light shown by Chitose et al65 . Kabra et al 66 reviewed the utilization of photocatalysis in the treatment of water contaminated by organic and inorganic pollutants. From their results, TiO2 nanoparticles degrade organic compounds e.g. chlorinated alkanes and benzenes, dioxins, furans, PCB etc. and also reduce toxic metal ions such as Cr (VI), Ag(I) and Pt(II) in aqueous solutions under UV light. Ashasi et al 67 synthesized N-doped TiO2 nanoparticles that were capable of photo degrading methylene blue under visible light. A wealth of information on TiO2 photocatalytic inactivation of bacteria has been acquired68. TiO2 can kill both Gram-negative and Gram-positive bacteria. More recently, nanosized TiO2 was also reported to kill viruses including poliovirus1 69, Herpes virus70, Hepatitis B virus71 etc. The antibacterial activity of TiO2 is related to ROS production, especially hydroxyl free radicals and peroxide formed under UV-A irradiation via oxidative and reductive pathways respectively72 . Nanomaterial Mechanism involved contaminants remediated Ref. Iron, Fe (0) precipitation, Immobilization, DNAPLs such as TCE, Organic [3,4, Sorption, Fenton’s reagent, chlorinated compounds, metals, 6,10] PRBs, Bimetallic metalloids, radionuclides etc. CNTs adsorption, filtration membranes metals, metalloids, organic and [14,15,18] SWNTs antimicrobial biological impurities etc. MWNTs FunctionalizedCNTs Silver, Ag (0) antimicrobial, disinfection bacteria, fungi, virii etc. [41] by generation of ROS TiO2 photocatalysis, oxidative organic compounds, toxic [47, 62,64]
Limitations of nanotechnology in water purification
Applications of nanomaterials are of definite advantage in many functional areas including water treatment. Although nanoparticles provide high specific surface area, a primary reason for their high reactivity, aggregation in water negates this benefit. Release of nanomaterials into the environment can have broader impacts on our ecosystem. It is therefore critical that researchers in this area to address questions such as ? What are the most environmentally benign methods for producing nanomaterials? ? What is the behavior of nanomaterials in environment and where these mostly end up in the environment? ? What are the toxic effects of nanomaterials when they interact with the organisms? Unfortunately, there is an insufficient data on the potential for accumulation of nanomaterials in environmentally relevant species and there have been few studies on the effects of many nanoparticles on the environmental communities. The properties that can be harmful to the environment are the very same properties that are advantageous and exploited during treatment and remediation processes. For instance, the catalytic properties of nanoparticles that induce the degradation of pollutants can also induce a toxic response when taken up by the cells. The factors and processes affecting ecotoxicity are complex, and the impact of manufactured nanoparticles on organisms is determined by a range of properties, including dissolution, aggregation, surface properties, the characteristics of exposure environment and the biochemical and physiological traits of the organism being exposed73 . Retention of nanomaterials is critical not only because of the cost associated with loss of nanomaterials, but also, and more importantly, because of the potential impacts of nanomaterials on human health and ecosystems74. The increased mobility of nZVI on its surface modifications would allow efficient remediation; it could also result in the possibility of nanomaterials migrating beyond the contaminated plume area, discharging to drinking water wells during the remediation process. Bulk TiO2 particles are known to be harmless to humans and animals. Although nanoscale TiO2 was classified recently as a possible carcinogen if inhaled75 its potential injestion via water is not expected to be a major concern, as reflected by its use in toothpaste and sunscreens. Researchers found that CNTs, if inhaled in large proportions could be as danger as asbestos. According to Lam and his co-workers, CNTs are light and could get air borne and when they enter into lungs, lesions were formed and toxicity greater than that of quartz was observed76 . The toxicity of well dispersed CNTs is less compared to CNT agglomerates77. In CNT agglomerates, the fraction of non-CNT soot like particles is higher than well dispersed CNTs which is believed to be one of the main reasons for increased toxicity of agglomerated CNTs. The negative health impact of Ag+ is darkening of skin and mucous membrane due to long term exposure to high silver concentration. Nevertheless, available information is insufficient to determine the highest allowable concentration of a particular nanomaterial in drinking water. The matter of determining the substance as dangerous involves not only in determining the material’s toxicity, but also to what degree the material will come into contact with the living cells. When materials are persistent and resist degradation, they may be present in the environment for long periods and have a greater chance of integrating with the living environment. Further research is needed to develop and understand the mechanism affecting the fate and transport of manufactured nanoparticles and their interaction with other organisms and how these interactions are influenced by different environmental variables. All these improvements can increase the ability to remediate more of the hazardous waste sites and minimize potential harm. CONCLUSIONS Emerging nanomaterials and the technologies will create tremendous opportunities for improvement and accessibility of water treatment technologies. The problems and challenges are multidisciplinary and begin with accurately understanding the potential for the environmental releases of nanomaterials and continue throughout characterizing environmental behavior, fate and bioavailability. In order to prevent any potential adverse environmental impacts, proper evaluation of these nanomaterials needs to be addressed before used on a mass scale. Future research addressing scalability, economics and safety of these systems is likely to overcome many of the current limitations and create opportunities to revolutionize drinking water treatment. ACKNOWLEDGEMENT The authors of this article greatly acknowledge the immense help received from the scholars whose articles are cited and included in the references of this review manuscript. The authors are also grateful to the authors, editors, publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesCYTOGENETIC EFFECT OF PHYTOPESTICIDE AGAINST LEPIDOPTERAN PESTS
English5459Maria Packiam SEnglish Elumalai KEnglish Ignacimuthu SEnglishThe present investigation aims to assess the cytogenetic effect of PONNEEM, a newly prepared botanical pesticide against the fourth instar larvae of Spodoptera litura Fab. PONNEEM exhibited spectacular changes in the morphological structure of chromosome of S. litura at 10 ppm concentration. The various karyomorphological changes namely metaphase plates showing clumping of chromosome, dots chromosome, endo reduplication, ploidy and deletion of chromosome were observed. This is the first report on the cytogenetic effect of PONNEEM against fourth instar larvae of S. litura. The severe damage on chromosome of S. litura was noticed due to the synergistic effect PONNEEM consisting of pongam and neem oils. So, this effective botanical pesticide which is safe and eco-friendly can be used as an alternative to chemical pesticides to control lepidopteran pests.
EnglishSpodoptera litura, PONNEEM, Karyomorphology, Ploidy, CytogeneticINTRODUCTION
Karanjin which is the active principle obtained from the plant Karanj or Pungam tree.[1] More than 19 biologically active components have been identified from this plant. Karanj is reported to be effective in controlling insect pests of stored grains, field [2] and plantation crops. It acts as oviposition deterrent or antifeedant or insecticide [3] against insect pests [4]. Neem tree is a versatile tree containing large number of chemicals and biologically active compounds. At present more than 100 triterpenoids and a few non-isoprenoid constituents like phenolics, carbohydrates and proteins and sulphur compounds have been isolated from neem tree. Azadirachtin A was the first member isolated by Butterworth and Morgan [5] from neem seeds showing the antifeedant and insect growth inhibitor effects on Schistoereca greagaria. Due to the presence of bitterness in different parts of neem tree, it has emerged as an important source as well as an alternative for the synthetic insecticides. It is known for its diverse effects like repellency, feeding deterrencey, oviposition deterrencey and growth inhibitory action on insect pests.[6] The combination of pungam and neem oils (1:1 ratio) was named as 'PONNEEM'. PONNEEM was patented (Indian Patent No. 204381) by the Entomology Research Institute, Loyola College, Chennai, Tamil Nadu, India) because of higher bioactivities against field insect pests [7] . This present investigation was aimed at finding the impact of 'PONNEEM' on the chromosomes of Spodoptera litura which is a polyphagous insect pest [8] damaging more than 120 plant species.
METHODOLOGY
Insect Rearing
Egg masses of Spodoptera litura were collected from groundnut field at Vellavedu village near Poonamallee, Chennai. The eggs were surface sterilized with 0.02% sodium hypochlorite solution, dried and allowed to hatch. After hatching the neonate larvae were reared on castor leaves till pre pupal stage and sterilized soil was provided for pupation. The pupae were collected from the soil and kept in oviposition chambers (40 x 25 x 25 cm). After adult emergence, cotton soaked with 10% (w/v) sugar solution with multivitamin drops was provided for adult moths to increase the rate of fecundity. Petioles of fresh castor leaves inserted in conical flask containing water (to avoid early drying of the leaves) were provided for egg laying. After egg laying, the egg masses were collected from the leaves and surface sterilized with 0.02% sodium hypochlorite solution, dried and allowed to hatch. The fourth instar larvae reared in the laboratory were used for the present study.
Preparation of PONNEEM
Pongum and neem oils were taken at 1:1 ratio in a stainless steel vessel with a stirrer and were stirred at 120 rpm for 10 minutes. Then 8% emulsifier and 1% stabilizer were added to the oils and again it was stirred at 120 rpm for 10 minutes. At last 0.123% Azadirachtin and 2% isopropyl alcohol were added and again it was mixed thoroughly by using a stirrer at 120 rpm for 10 minutes. Then PONNEEM was obtained [9,10] .
Chromosomal Analysis
The larval sex organs (ovary and testes) of 10 ppm PONNEEM treated and control larvae of S. litura were dissected out in 0.001% Colchicine solution and were left in the same solution for 2.5 hours and then they were transferred to hypotonic solution (0.45% sodium citrate solution). After 30 minutes, it was replaced by fresh hypotonic solution and kept at room temperature for 30 minutes. At the end of 30 minutes, the tissue was transferred to a clean dry slide and a drop of acetic acid: methanol (1:3) mixture was added and squashed for a minute. Then the slides were freezed on dry ice for 2-3 minutes, thawed and kept in glacial acetic acid for 30 seconds and subsequently air dried for 45 minutes. The air dried slides were stained in Giemsa, diluted 30 times in Sorrenson’s phosphate buffer having pH 6.8, for 15 minutes at room temperature and then excess stain was de-stained in double distilled water and then the slides were air dried. The air dried slides were observed on the next day (Murakami and Imai, 1974)[11]. The various chromosomal aberrations in treated larvae of S. litura were observed under the microscope.
RESULTS
Damage on chromosome of S. litura
Effect of PONNEEM at 10 ppm concentration against larvae of S. litura was apparent in view of its cytogenetic effects. Fourth instar larvae exposed to such concentration were screened for chromosomal aberrations. Results clearly revealed that PONNEEM had significant effect in altering the genetic constitution of the target insect pest carrying reduction in the growth of larvae when compared to the untreated larvae (Figure 1).
Chromosomal aberrations like clumping of chromosome (Figure 2: A), condensation into dots (Figure 2 : B) were seen. Many metaphase plates showed chromosomal variation in number, though they had distinct morphology (Figure 2: C & D). Ploidy was the common phenomenon observed in all the experimental larvae (Figure 2: F).
DISCUSSION
Fourth instar larvae exposed to such concentration of phytopesticidal formulation have been screened for chromosomal aberrations. Several damages on chromosomes of S. litura were observed at 10 ppm concentration of PONNEEM. Chromosomal aberrations like clumping of chromosome, condensation into dots were seen. Many metaphase plates showed chromosomal variation in number, though they had distinct morphology. The present finding coincides with the findings of Maheswaran and Ignacimuthu [12] who noticed that PONNEEM showed remarkable ovicidal activity of against human vector mosquitoes Aedes aegypti and Aedes albopictus. Similarly Packiam and Ignacimuthu [15] noticed that PONNEEM treated larvae of S. litura had reduced egg hatchability when they emerged into adults. In the present investigation, PONNEEM exhibited the damage on chromosome of S. litura because of the synergistic effect of active compounds like azadirachtin (Fig. 3) and karanjin (Fig. 4). In an earlier report, neem oil based formulation of active fraction of ethyl acetate extract of Hydnocarpus alpine showed antifeedant activity against S. litura [16]. Pavunraj et al., [17] reported that effective fraction from Melochia corchorifolia with 1:1 ratio of neem and pongam showed antifeedant activity against four lepidopteran pests.
CONCLUSION PONNEEM
produced high damage on the chromosome of S. litura. It was highly toxic to lipodopteran insect pests in which internal cells were damaged, Its normal growth was arrested due to its synergistic activity. This is the first report on the cytogenetic effect of PONNEEM against S. litura. This effective phytopesticidal formulation can be a safe and alternate to chemical pesticides. This is also good for integrated pest management programme in agriculture.
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=1995http://ijcrr.com/article_html.php?did=19951. Venugopala Verma SR, Gupta SP, Tyagi GP. Studies on the toxicity of lindane on Colisa fasciatus. Part I. TML measurement and histopathological changes in certain tissues. Gegenbaurs Morp Jahrp Leipzic. 1975; 121(1):38-54.
2. Kumar V, Chandrashekar K, Sidhu OP. Efficacy of karanjin and different extracts of Pongamia pinnata against selected insect pests. J Entomol Res. 2006; 30:103–108.
3. Kabir KE, Islam F, Khan AR. Insecticidal effect of the petroleum ether fraction obtained from the leaf extract of Pongamia glabra Vent. on the American cockroach, Periplaneta americana (L.) (Dictyoptera: Blattidae). Int Pest Control. 2001; 43:152–154.
4. Mukesh Kumar, Ram Singh M, Kumar, Singh R. Potential of Pongamia glabra vent as an insecticide of plant origin, Biological Agriculture and Horticulture. 2002; 20:29.
5. Butterworth JH and Morgan ED. Investigations of the locust feeding inhibition of the neem tree Azadirachta indica. J Insect Physiol. 1968; 17:969-977.
6. Harikrishnan R, Rani MN, Balasundaram C. Hematological and biochemical parameters in common carp, Cyprinus carpio, following treatment for Aeromonas hydrophila infection. Aquacult. 2003; 221: 41-50.
7. The patent office journal dated on 19/05/2006, visited on 20/07/2012 at http://www.tmpsearchers.com/Patdatabase/20 06May19/pg_0021.htm.
8. Baskar K, Antony Raj G, Murali Mohan P, Lingathurai S, Ambrose T, Muthu C. Larvicidal and Growth Inhibitory Activities of Entomopathogenic Fungus, Beauveria bassiana against Asian Army Worm, Spodoptera litura Fab. (Lepidoptera: Noctuidae). Journal of Entomology. 2012; 9(3):155-162.
9. Packiam SM, Ignacimuthu S. Effect of PONNEEM on Spodoptera litura (Fab.) and Its Compatibility with Trichogramma chilonis Ishii. Braz Arch Biol Technol. 2012; 22: 291- 298.
10. Packiam SM, Baskar, K, Ignacimuthu S. Ovicidal activity of botanical oil formulations against Helicoverpa armigera Hubner and Spodoptera litura Fabricius (Lepidoptera: Noctuidae). Asian J Tropical Biomed (In press). 2012.
11. Murakami A, and Imai HT. Cytological evidence for holocentric chromosomes of Bombyx mori L. and Bombyx mandarina M. (Bombycidae), Chromosoma, 1974; 47(2):167-178.
12. Maheswaran R, Ignacimuthu S. A novel herbal formulation against dengue vector mosquitoes Aedes aegypti and Aedes albopictus. Parasitol Res. 2012; 110:1801- 1813.
13. Butterworth, J.H. and Morgan, E.D. 1968, Isolation of a substance that suppresses feeding of locusts. Chemical Commun. 23-24.
14. Vismaya, Sapna Eipeson W, Manjunatha JR, Sriniva P, Sindhu Kanya TC. Extraction and recovery of karanjin: A value addition to karanja (Pongamia pinnata) seed oil. Industrial Crops and Products. 2010; 32(2): 118-122.
15. Packiam SM, Ignacimuthu S. ibid.
16. Vendan SE, Lingathurai S, Paulraj MG, Ignacimuthu S. Bioefficacy of neem oil formulation with Hydnocarpus alpina leaf extract against Spodoptera litura. Int J Curr Res. 2010; 3:78-82.
17. Pavunraj M, Baskar K, Ignacimuthu S. Efficacy of Melochia corchorifolia L. (Sterculiaceae) on Feeding Behavior of Four Lepidopteran Pests. Int J Agric Res. 2012; 7: 58-68.
A. Metaphase plates showing clumping of chromosome
B. Photomicrograph showing dots chromosome, endo reduplication and ploidy nature
C. Metaphase plate showing rods and dots like chromosomes. Arrow indicates deletion of chromosome. Encircled chromosome indicates attachment of a fragment (addition of chromosome)
D. Metaphase plate showing rods and dots like chromosomes. Arrow indicates deletion of chromosome. Encircled chromosome indicates attachment of a fragment (addition of chromosome)
E. Ploidy Chromosomes with distinct morphology
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcarePOSTMORTEM STUDY OF FRACTURE OF NECK STRUCTURES IN HANGING AND LIGATURE STRANGULATION DEATHS
English6065Sadikhusen G. MominEnglish Sanjeevkumar ChoudharyEnglish Hetal C. KyadaEnglish Dipak H. VoraEnglishBackground: The Hyoid bone is the most important part of internal examination at the autopsy table while performing autopsy in cases of Hanging and Ligature strangulation. Fracture of hyoid bone has been ascribed to many factors like manners of constriction, level of application of ligature material or force of constriction, long drop or short drop suspension, age and sex of victim etc. It is also very important to check whether it is ante mortem or postmortem in nature or just an artifact. Observing the importance given to hyoid bone fracture in hanging and ligature strangulation cases by many authors in past and present days, the present prospective study was performed on asphyxial deaths due to hanging and ligature strangulation. Objectives: To study the fracture of neck structures in hanging and ligature strangulation deaths. Study Design: Prospective study. Study Setting: This study was conducted at P.D.U. Medical College and Hospital, Rajkot during January 2008 to December 2008. Result: During that period out of 2159 cases, 90 cases of hanging and 7 cases of ligature strangulation were selected for the present study. Hyoid bone was fractured in 16 cases (17.78%) of hanging. In one case (1.11%) of hanging where thyroid cartilage was fractured. In two cases (28.56%) of ligature strangulation, thyroid cartilage was fractured. No fractures of the cricoid cartilage or cervical spine were demonstrated. Conclusion: Incidences and postmortem findings of fracture of neck structures in cases of hanging and ligature strangulation were almost same which was observed by the other recognized authors in the field.
EnglishHanging, Ligature strangulation, Fracture of Hyoid bone, Fracture of Thyroid cartilageINTRODUCTION
Osteocartilagenous structures of neck like hyoid bone, thyroid cartilage, tracheal cartilages and cervical vertebrae show injury with variable incidences in cases of hanging and ligature strangulation. Hyoid bone is commonest to be injured in conventional hanging cases, whereas fracture or dislocation of cervical vertebrae may be noticed in judicial hanging where the victim jumps from a height and his fall is arrested by a sudden jerk of ligature. The fracture of hyoid bone depends on age of individual and type of compression. After the ossification of greater cornu with body after 40 years there is loss of mobility and in turn variation in incidence of fracture of hyoid bone in case of hanging has been reported by different workers i.e. 4.9% by Patel et al1 , 15.38% by Gargi et al2 , 9.4% by Momonchand et al3 and 5.1% by Sheikh et al4 . The type of hyoid bone fracture (if present) can help to decide the manner of neck compression hanging or strangulation (ligature or manual). Commonly, the lateral compression fracture is seen in strangulation and anteroposterior fracture of hyoid bone suggests hanging. However, whether the each fractured piece displaced inward or outward and opinion about type of neck compression needs careful observation and expert interpretation5 .
MATERIAL AND METHOD
This prospective study was conducted in all cases of death due to hanging and ligature strangulation for the purpose of studying the fracture of neck structures at the Department of Forensic Medicine, P.D.U. Medical College and Hospital, Rajkot from January 2008 to December 2008. During that period out of 2159 cases, 90 cases of hanging and 7 cases of ligature strangulation were selected for the present study. A detailed history from police and relatives regarding age, sex, socio economical status, marital status, habits, illness (mental / other disease / deformity), previous attempted suicides, suicide note if any etc. were taken. Detailed history from police regarding scene of crime, position of body etc. were taken. Irrespective of information collected, both external and internal post mortem findings were observed meticulously. Neck structures were inspected carefully; neck dissection was conducted for presence or absence of fractures of bony structures of neck. The tongue, larynx and trachea were removed carefully. Then the findings were confirmed by meticulous dissection to observe hemorrhage in and around the fractured area.
OBSERVATIONS
Total number of 2159 cases was brought for postmortem examination during the period from 01-01-2008 to 31-12-2008, out of which 97 cases (4.49%) of hanging and ligature strangulation were studied. Maximum 39 cases (40.2%) of hanging and ligature strangulation deaths were reported in age group of 21-30 years. Out of 36 cases, 24 (64%) male and 12 (34%) female died due to hanging while 3 case of ligature strangulation equal reported in male. Chunni was used as a ligature material in maximum 31 cases (34.44%) of hanging followed by nylon rope in 28 cases (31.1%) whereas in ligature strangulation cases, ligature material was not known in maximum number 3 cases (42.85%). As per Table-3, in all cases of hanging only one ligature mark was present. The ligature mark was situated above thyroid cartilage in 72 cases (80%) of hanging. Ligature mark was one in number in 6 cases (85.71%) of ligature strangulation, in all 7 cases (100%) it was situated below thyroid cartilage. As per Table 4, injuries to osteocartilagenous structures of neck were highest in hanging cases in age group of 31 to 40 years followed by 21 to 30 years age group. Incidence is increasing with increase in age. In 90 cases of hanging, hyoid bone was fractured in 16 cases (17.78%). Out of 16 cases, 11 cases (12.22%) were male and 5 cases (5.43%) were female. While in one case (1.11%) of hanging, thyroid cartilage was fractured and in 2 cases (28.56%) of ligature strangulation, thyroid cartilage was fractured. Hyoid bone was not fractured in any case of ligature strangulation. Larynx, trachea and cervical vertebra were not fractured in any case of hanging and ligature strangulation.
SUMMARY AND CONCLUSION
1. In the present study 4.49% cases were declared on autopsy of hanging and strangulation deaths which is similar to study of Sheikh et al4 .
2. In the present study maximum number of cases (40.2%) were reported in the age group 21-30 years, which is consistent with observations of Sheikh et al4 (42.4%) and Joshi et al7 (44.18%). 21-30 years age group involves maximum because they are more vulnerable to the fast changing social trends and cultures, as they are mentally a bit immature with little experience of life and there is great fluctuation of emotion in this age group.
3. In hanging cases 66.67% male and 33.33% female cases were observed in present study, which is consistent with observations of Sheikh et al4 and Jani et al6 . This could be due to more activeness of male in society and less carelessness in females. Males are more active in various activities and customs and hence they are vulnerable for more stress and tension.
4. Chunni as a ligature material was used in 34.44% cases of hanging in the present study. In study by Sharma B R et al8 commonest ligature material was chunni in 17 cases (30.90%). The reason for selecting chunni can be explained on the basis of easy availability in Indian house.
5. Number of ligature mark is one in all cases of hanging are similar with observation of Momonchand et al3 (96.7%). In present study six case of ligature strangulation where number of ligature mark is one while in one case, number of ligature mark is more than one. Ligature mark was situated above the level of thyroid cartilage in 72 cases (80%) of hanging which is similar with observation of Naik S K9 (82.94%). In all 7 cases (100%) of ligature strangulation, ligature mark was found below thyroid cartilage. 6. Incidence of fracture of hyoid bone in present study (17.78%) is almost similar with Simonsen10 (17.5%) and Jani et al6 (17.4%). 7. Incidence of injuries to osteocartilagenous structure of neck in hanging cases increases with increase in age. The observations of present study are consistent with the work of other workers.6,11,12 8. The percentage of hyoid bone fracture in hanging deaths vary greatly from 0% to 68% from author to author when we look into different text like Wintraub (1961) reported in 27% cases of hanging13, Betz P. and Eisenmenger W (1996) reported Throat skeleton fracture seen in 67% hanging14 , Nikolic S, Micic J, Antanasijevic T, Djokic V. and Djonic D(2003) reported Throat skeleton fracture seen in 68% hanging15 and Reddy K.S.N.(2008) seen in15-20% cases of hanging but rare below 40 years of age5 .
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=1996http://ijcrr.com/article_html.php?did=19961. Patel H R and Kothari D R. A study of asphyxial deaths from 1985 to 1989 at a hospital at Ahmedabad. The special Conference Issue of Indian Academy of Forensic Medicine, Calcutta 1992; 107-114.
2. Gargi J, Gorea R K, Chanan A and Mann G. Violent asphyxial deaths- a six year study (1985-1991). The special Conference Issue of Indian Academy of Forensic Medicine, Calcutta 1992; 171-176.
3. Momonchand A, Meeradevi T. H. and Fimate L. Violent asphyxia death in Imphal. Journal of Forensic Medicine and Toxicology (1998); 15(1):60-64.
4. Sheikh M I and Agarwal S S. Medicolegal implications of hyoid bone fracture- a study paper. Journal of Indian Academy of Forensic Medicine. (2001); 23(4):61 – 63.
5. Reddy K S N. The Essential of Forensic Medicine and Toxicology, 27th edition, K. Saguna Devi. Hyderabad (2008); 313-14.
6. Jani C B and Gupta B D. An autopsy study of parameter influencing injury to osteocartilagenous structures of neck in hanging. International Journal of Medical Toxicology and Legal Medicine (2002); 5(1):4-7.
7. Joshi Rajeev, Chanana Ashok and Rai Hakumal. Incidence and Medicolegal importance of Autopsy study of fracture of Neck structure in hanging and strangulation. Medico legal update (2007); 7(4):105-109.
8. Sharma B R, Harish D and Singh V P. Ligature Mark on neck: How informative? A study paper Journal of Indian Academy of Forensic Medicine (2005); 27(1): 10 -15.
9. Naik S K. Obliquity vs. discontinuity of ligature mark in diagnosis of hanging – A comparative study. Anil Agarwal internet journal of forensic medicine and toxicology 2006; 7(1). Jan -June.
10. Simonsen, Jorn. Patho-anatomic findings in neck structures in asphyxiation due to hanging- a survey of 80 cases. Forensic Science International (1988); 38:83 – 91.
11. Paparo, Gary P and Siegel Henry. Neck markings and fractures in suicidal hangings. Forensic Science International (1984); 24:27- 35.
12. Morild I. Fracture of Neck structure in suicidal hanging. Medicine Science and Law (1996); 36 (1):80 – 84.
13. Weintraub CM. Medicolegal Journal (Camb). (1961); 21:209-16.
14. Betz P and Eisenmenger W. Frequency of Throat-Skeleton Fracture in Hanging. American Journal of Forensic Medicine and Pathology (1996); 17(3):191-193.
15. Nikolic S, Micic J, Antanasijevic T, Djokic V, Djonic D. Analysis of Neck Injuries in Hanging. American Journal of Forensic Medicine and Pathology (2003); 24(2):179-182
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN-0001November30General SciencesSUPPLY CHAIN MANAGEMENT IN SUGAR INDUSTRY: A STUDY OF WESTERN MAHARASHTRA IN INDIA
English6670N C DhandeEnglish V.R.SalkuteEnglishAgro processing is a large sector that covers activities such as agriculture, horticulture, and plantation. It also includes other industries that use agriculture inputs for manufacturing of edible products. Sugar Industry is one such major player in Agro processing sector. Greater delegation and competitiveness in agro based industries require new forms of co-ordination between farmers and other agencies to increase quantity within catchment area. During the entire production season, to maximize sugar production with minimum overheads a good network of all the elements is required to be in place. Sugar industry faces a lot of problems such as lack of co ordination, interruptions in supply of raw material due to natural calamities and many other such problems. To overcome these problems industry needs to have a proper supply chain management. Sugar industry particularly in Maharashtra is very popular in the cooperative sector. It is contributing to nearly @ 40% of total national production of sugar. The tonnage figures are @ 50-70 lakh tons of sugar every year[2].
EnglishSugarcane, supply chain Management, sugar Industry, production.INTRODUCTION
Sugar industry constitutes one of the most important agro based industries in India. It started growing in an organized way during the 1930 after introduction of sugar industry protection act in 1932. Sugar industry in India is well developed with a consumer base of more than billions of people. It is also the second largest producer of sugar in the world. The Indian sugar industry has a turnover of Rs. 500 billion per annum and it contributes almost ` 22.5 billion to the central and state exchequer as tax cess and excise duty every year (ref: Ministry of food, Government of India report). Sugar industry directly provides employment to 45 million i.e. 7.5% of rural population engaged as sugarcane farmers, agriculture laborers and dependents. It also employs 0.5 million skilled and semi skilled workers. In aggregate employment generated by sugar industry is for more than Five million persons. It is considered as engine of rural development and socio economic upsurge. In the sugar industry management of supply chain is changing considerably in response to the often difficult market environment. Market forces present real challenges to wholesalers and retailers but are exceptionally problematic. for primary producers who often lack the knowledge to manage these new and dynamic relationships. Improving efficiency of supply chain is now become the need of the hour as this industry has to compete at international level. Although India’s average farm size is smaller, the industry maintains that smaller farmer is a valuable part of the grower base and will continue to be so into the immediate future. This characteristic underlies the importance of initiatives such as sharing of farm equipment and harvesting by grower owned cooperatives or partnerships as well as the need to support farmer’s diversity of income sources[1][2].
Supply chain management in sugar industries involved a lot of problems which will in turn affect the productivity of sugar. The sugar industries in India use only sugarcane as input, hence sugarcane have been established in large sugarcane growing states like Uttar Pradesh, Maharashtra, Karnataka, Gujarat and Tamilnadu and Andhra Pradesh. These six states contribute more than 85% of total sugarcane production in India. Uttar Pradesh and Maharashtra together contribute more than 57% of total production [3].
FACTORS AFFECTING SUPPLY OF SUGARCANE
-Climate- Due to global warming there are frequent changes in the climate which in turn affect the yield and quality of sugarcane
- Technical- Lack of technical knowledge among farmers regarding cultivation and harvesting of sugarcane - Sugarcane production
- When there is more production of sugarcane it will lead to more production of sugar and vice versa.
- Political factors- This is major factor affecting the supply of sugarcane as majority of sugarcane industries in western Maharashtra are dominated by politicians and the political battle strongly affect this industry.
- Recovery rate, Duration of season, sugarcane utilization for sugar production is also another factor that affects the supply of sugar[1].
Structure of sugar industry in India
Public: 06%
Private: 54%
Cooperativ: 40%
MAJOR PROBLEMS IN SUPPLY CHAIN OF SUGAR INDUSTRIES
A) CANE PRODUCTION
In the region, cost of production- including harvesting, infield haulage, road transport, loss of throughput can add to costs at every link in the supply chain value chain research provides some initiatives for reducing this cost and improving the efficiency of supply chain, like the implementation of GPS guidance system to assist tracking while harvesting, group farming, rationalization of harvesting group to reduce cost of growers, upgrading the website to facilitate the collection and analysis of data from farmers to assist with agronomic and farm financial management decisions, regularly providing the latest updates of market and climatic changes as well as the best practices for improving sugarcane production to the farmers by conducting workshops and special training programme will also help to improve the production of sugarcane.
B) TRANSPORTATION
Sugarcane industries are mostly located in rural areas as sugarcane is the main raw material used for manufacturing of sugar so the sugar industries in order to reduce their transportation cost prefer to set up the industries near the of sugarcane production areas, but the main problem here arises is that improper road facilities. In absence of dynamic vehicle scheduling, a typical sugarcane transport system symptomatically exhibits excessive queuing times large number of no cane stops, significant over fleeting and poor vehicle utilization, a high level of coordination is required between farmers, harvest contractors, transport contractors and mill managers to make sure the mill receives a steady stream of fresh cane. The development of larger cane bins and an efficient trapping system for the efficient transport of whole crop material will help to reduce the transportation cost to some extent.
C) WAREHOUSING
After manufacturing and before distribution the sugar is stored in warehouses which is also called as inventory, during this storage some problems may occur like sometimes due to moisture sugar get converted in solid cubes or may be spoiled due to improper handling. So proper care should be taken to maintain the inventory in good condition. To solve these problem industries should develop warehousing at consuming destinations this enables industries better pricing and eliminating too many middlemen, better Information Technology and application of quantitative tools and techniques can be used for inventory management.
GOVERNMENT POLICIES
Sugar has historically been classified as an essential commodity and has been regulated across the value chain. The heavy regulation in this sector artificially impacts the demand supply forces resulting in market imbalance. Sensing this problem, since 1993 the regulations have been progressively eased. The key regulatory milestones
include de-licensing of the industry in 1998 and the removal of control on storage and distribution in 2002 .However, policy still plays an important role in industries. There are various legislations regarding sugarcane procurement, sugarcane pricing, sugar sales, capacity and production, import and export.
OTHER PROBLEMS
Mismanagement, lack of coordination between farmers, mill owners and distributors, lack of modern technologies as many of the industries still rely on old technologies are some problems which need to be corrected for improving the efficiency of supply chain.
Problem of Farmers
- A marginal Indian farmer is faced with several challenges that result in yield and quality being inferior by world standards.
- He lacks access to information accurate weather forecasts and up to date information on commodity market prices.
- He lacks access to the market and thus caught in the vicious cycle of suboptimal use of farm inputs higher cost of credit and lower price realizations on his produce leading to indispensible income for him[1].
Solution for these problems
- Pre configured SAP solution
- Web based cane management system
- Add on utilities; smart card and SMS
- ASM clean development Mechanism.
CONCLUSION
In the sugar industry management of supply chain is changing considerably in response to the often difficult market environment. Geographic Information System(GPS) and Global positioning system(GPS) are used extensively in location, site selection, land use planning, environment science, transportation system, trucking companies now track their tracks via GPS technology. In vehicle navigation system, vehicle location system, emergency vehicle deployment and traffic management are using GIS and GPS for their value chain. In addition to this effective communication throughout the world had had communicated virtual world of e business, e integration, e commerce, e marketing, e procurement. Modern sugar industries are required to adopt all these measures for improving the efficiency of their supply chain management.
Englishhttp://ijcrr.com/abstract.php?article_id=1997http://ijcrr.com/article_html.php?did=19971 Problem and prospectus of sugar industry: a report from Agrovan daily news paper.
2 www.maharashtra rajya sakharsang
3 www.sugartech.coza/news/index
4 Indian agriculture ministry report July 2009
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareNEW TECHNIQUE FOR PREPARATION OF DRY SPECIMEN USING DISCARDED CADAVERIC PARTS
English7177Lalit Kumar JainEnglish Pooja Rajendra GangradeEnglish Neha VijayEnglishBackground and objectives: Anatomy is best learned through dissection, prosected parts, models, photos, computer simulations and various imaging modalities. Gross anatomy specimens too, area valuable aids, in the teaching of anatomy, but their storage and handling poses a great inconvenience due to the routinely used formaline preservation technique. Various scientists and anatomists have tried to preserve such specimens, some very expensive, some very complicated, some requiring skilled expertise. In the present study we have tried to come out with a new technique to make teaching specimens. Method: The procedure is far easier to pursue and moreover made from discarded specimens. Conclusion: The specimens thus made saves us from costly expenses, fuming formaldehyde and which are also student friendly as they can learn even from freely feeling and touching the specimen.
Englishformaline, museum techniques, plastination, specimens, teaching aidsINTRODUCTION
Museums have been historically the centres for education, creative thinking, virtual learning, and research. Visualisation of three dimensional specimens locked inside the jars of museums invite the students interest towards the part displayed. Significant advancement in the teaching methodology and the study of anatomy has been made in recent times. The credit goes to all the recent and past anatomists, who have from, time to time evolved newer and more efficient tools of imparting powerful knowledge to students. Anatomy is best learned through dissection, prosected parts, models, photos, computer simulations and imaging modalities like x- ray, cat scans in recent times. Prior to the advent of such technologies, many medical students did not have access to cadavers or prosected parts. To ease them of the difficulty some expert along with artists and sculptors created amazingly realistic renditions of various parts of human body based on observation of dissected cadavers. These models were first used in the late 17th century and were mainly made up of wax and came from Italy, they soon ceased to exist as they were fragile and very delicate to handle. Then came the era of plastic and metal, all these models posed a similar problem that they all were made by people who were not from medical background and with very slight or negligible knowledge of the human body, thus the outcome of such practise was not as fruitful as expected. It is well said,“Seeing promotes understanding and understanding in turn promotes the most practical kind of body education possible”. Although, the use of artificial tools like wax models, plastic and metal models help the learner, the importance of original body parts cannot be neglected, as the models are idealized body parts through the eyes of an artist. Gross tissue specimens are a valuable aid, in the teaching of anatomy and pathology, but their storage and handling poses great inconvenience because of the use of formaline. These are traditionally packed in glass jars filled with 10% formaline which makes it difficult to handle as they are fragile and due to the fluid the viewing becomes difficult. On removal from the jar, the practise becomes more unpleasant due to the formaldehyde fumes, also the exposure of these specimens to air, brings quick changes in their colour and deteriorates the features progressively. To avoid these barriers some scientists like Harold Meryman used the technique of freeze drying but it also had limitations as the architecture of the specimen was distorted1 .There were others who used forced air impregnation method2 . In 1977, a breakthrough by scientist and anatomist, Dr Gunthen Von Hagens, came into the world of specimen preservation technique which was termed as the plastination3,4. It is a process which halts the decomposition process by extracting all bodily fluids and soluble fats from the specimens, replacing them through vacuum forced impregnation with reactive resins and elastomers and then curing them with light, heat and certain gases which gives the specimen rigidity and permanency. But the process is extremely expensive and requires expertise in making the specimen. To avoid the expenses and lessen the burden of the expertise required, in our present study we have tried to make specimen from the unwanted, visibly distorted and discarded body parts, which nevertheless would have to see the face of an incinerator. MATERIALS AND METHOD Discarded dissected cadaveric parts were collected from the Dept. of Anatomy, at Geetanjali medical college, Udaipur (Rajasthan), India. After detailed discussion, on what best deep dissection could be carried out on the said parts the process began: All the dissected parts were cleaned to remove any excess connective tissue either on the muscles , tendons , blood vessels , nerves , even the bone was cleaned completely to remove any extra trace of muscle , tendon attachment and even stripped off of its periosteum. The joint cavity once opened and the muscle and neurovascular bundle to be showed in the specimen, were clearly demarcated by removing all the other tissues. the joint cavity and other tissues were then packed with cotton and covered adequately so as to preserve the in situ architecture (Fig 1). The specimen was then left in shade to dry for at least a week. After inspection of the specimen that it had dried, the neurovascular bundles and adjoining ligaments or tendons or muscles were coloured using the anatomical scheme of colours , i.e. yellow for nerves, red for artery, blue for veins, green for ligaments and tendons, using a paint brush and oil colours (Figs 2,3,4,5,6,7). After leaving the specimen to dry again the parts were then prepared to be polished by series of coats of varnish. This coat protects the bone from any destruction by the environmental factors and gives the lustre to the soft tissue and hard parts. The mounting of the specimen was also done using discarded material, for which an empty 2 litre soft drink bottle, a thin steel rod, some pebbles and plaster of Paris were required. The bottom of the soft drink bottle was cut from above horizontally leaving around 10 cm of it from its base, this would act as a cast for making the stand. A vertical slit was made on the above cast, so that it could be opened easily. A mixture of plaster of Paris was made and a steel rod was placed in the centre of the cast. Along with the plaster of Paris, small pebbles were put in the cast, these pebbles would give the base more weight and make it more stable and allowed to set. Later the cast was removed and the base was coloured using desired scheme (Fig 8). The bone was drilled in its lower end and the steel rod fixed in it. DISCUSSION The above approach, used to make dry specimens using the discarded parts of the human body have many advantages over the past methods described above5,6 . At the onset, the parts are readily available in the department, like other procedures we do not have to depend on sources from outside which are usually very expensive, like the ones used in the process of plastination where resins and polymers are used, formalin fixed jars where formaldehyde is required. Furthermore the parts used are almost discarded after dissection by students, thus we do not use new cadaver or a fresh body part to make the specimen. The process is very cost effective, colours, plaster of paris are very cheap resources, easily available and usually are present in all departments. No special expertise is required as seen in case of plastination, where only the expert can perform the procedure under conducive environment and well established laboratory. Here even a postgraduate or an undergraduate student can make a specimen under guidance, thus can be made at every teaching institute. Due to the nature of the specimen, it can be handled easily, without the fear of either spilling the formalin or breaking the model. The architecture of the specimen is maintained truly to its original state, and students can visualise the same by touching and feeling the structures and without any refractive media disturbance , which can make the study cumbersome . The normal colours of tissues are maintained and specimens are more durable as they can last longer. These can be easily transported and used as teaching aids even in lectures and demonstrations.
CONCLUSION
Although the above study has several advantages to it, yet it has its own limitations. as the study hasn’t been performed on bodily viscera and other soft tissues till date . But it has a promising future in the field of anatomy and museum techniques.
ACKNOWLEDGEMENT
Authors acknowledge the immense help received from the scholars whose articles are cites and included in the references of this manuscript. The authors are also grateful to the 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=1998http://ijcrr.com/article_html.php?did=19981. Harold TM. Freezing and drying technique of biological materials: Annals of The New York Academy Of Sciences 1960, 85:503- 734.
2. Henry RW, Butler J. Room temperature “forced air impregnation” off dried lung with S10/S3 xylene mix: J Int Soc Of Plastination 1990, 4: 14-15.
3. Von Hagens G, Tiedemann K, Kriz W. The current potential of plastination: Anat Embryology 1987, 775(4): 411-421.
4. Nicaise M, Simones P, Lauwers H. Plastination of organ: A unique technique for preparation of illustrative demonstration specimens: Flemish Veterinary Journal 1990, 59: 141-146.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesMICROBIAL DEGRADATION OF HYDROCARBONS FROM OIL CONTAMINATED SOIL BY USING PSEUDOMONAS SP.
English7885B. KamaladeviEnglish Prabhavathi. PEnglish Sankareswaran. MEnglishAims: To study the microbial degradation of hydrocarbon from oil contaminated soil sample using Pseudomonas sp. and were analyzed the different physicochemical characters of normal and oil contaminated soil. Method and Results: The soil sample was collected from oil contaminated site, in Sivakasi and then the oil degrading microorganism (Pseudomonas sp) was screened from oil contaminated soil by using enrichment technique. After, the isolated microorganisms were identified by using standard characterization method. From the oil contaminated soil, 10 bacterial strains were isolated. Among them, tolerance of two strains was used for further studies. The physico-chemical characters of normal and oil contaminated soil samples were analyzed by standard method. In this study, the pH, moisture, carbon, phosphorous and potassium content of normal and oil contaminated soil samples were analyzed and compared. In the present investigation, the various concentration of aromatic hydrocarbon (toluene) was degraded by Pseudomonas sp and rate of the degradation was analyzed by HPLC (High Performanace Liquid Chromatography). The chemo taxis properties of the Pseudomonas aeruginosa and Pseudomonas fluorescens were studied by chemo taxis assay and also studied biosurfactant production of Pseudomonas sp. simultaneously the biomass, emulsification activities of Pseudomonas sp on toluene was studied. This biosurfactant production was detected by Thin Layer Chromatography. Conclusions: In this present investigation, the aromatic hydrocarbon degradation efficiency, chemotaxis properties and biosurfactant production of Pseudomonas sp. was compared with MTCC cultures of Pseudomonas stutzeri. Significance and impact of the study: Treatment with adapted bacteria could degrade the hydrocarbons.
EnglishOil degrading microorganism, Thin layer chromatography, Biosurfactant.INTRODUCTION
As we dig deeper into the modem industrial age technologies, several aspects of human life change. People benefit largely from life development and many live in prosperity, but property has price. This price is paid by our environment that suffers daily from pollutants and destruction. People now have to find ways to cure this destruction. One of the most destructive pollutants known today is oil contamination. Oil contamination is one of the most dangerous pollution factors. It can cause a threat to the environment. It is very feared by environmentalists and it’s very hard to control if it gets out hand. Oil spillage still occur through tanker accidents, well blow out, Sabotage and accidental rupture of pipelines, resulting in the release of crude and refined oil in terrestrial and aquatic environments (Atlas, 1981; Colwell and Walker, 1977). Oil contamination can be either at sea or in soil level. Sea oil contamination poses a threat on marine life and the death of hundreds of marine organisms. Soil contamination on the other hand is slow and lethal. It acts over a long period of time by poisoning food crops, water supplies etc. Oil contamination in soil results in an imbalance in the carbon-nitrogen-phosphorus ratio in the spill site, because crude oil essentially a mixture of carbon and hydrogen. This cause a nitrogen deficiency in oil soaked Soil, which retards the growth of bacteria and utilization of carbon sources (Regina et al., 2006). Furthermore, large concentrations of biodegradable organics in the top layer deplete oxygen reservoirs in the soil and slow down the rate of oxygen diffusion into deeper layers. Toluene is widely as an organic solvent, and its world wide production is estimated to be more than 80,000 metric ton per year. Toluene and other solvents, such as xylene, benzene and ethyl benzene are ubiquitous pollutants (Atlas, 1995) and several environmental protection agencies have declared the removed of there pollutions to be a high priority. In many environments indigenous microorganisms are able to remove aromatic hydrocarbons. This is to be expected, as many catabolic pathways for the metabolism of these compounds have been described, particularly in strains belonging to the genus Pseudomonas. Aromatic hydrocarbons are extremely toxic for microorganisms because they dissolve in the cell membrane (Sikkema et al., 1995). Therefore a critical issue in the degradation of aromatic hydrocarbons in polluted sites is tolerance to these compounds. Recently a number of Pseudomonas strains have been shown to be tolerant to organic solvents such as Toluene, Xylene, styrene and others. Solvent tolerance in bacteria belonging to the genes Pseudomonas involves (industry) an increase in cell membrane rigidity as a result of increase in both the level of the trans isomers of unsaturated fatty acid and level of cardio lipid a component of phospholipids head groups (Ramos et al., 1997), and oil removed of solvents from membranes via efflux pumps (Isken and Bont, 1996; Ramos et al., 1995). Bioremediation methods are currently receiving favorable publicity as promising environmentally friendly treatment technologies for the remediation of hydrocarbons (Desai and Banat, 1997). Biodegradation enables oil to be converted from harmful organic substances in to harmless inorganic substance (Zhao and Zhu, 1993). Three major factors affects the biodegradation of crude oil and these are chemical composition of oil, dissolved oxygen content of water and nutrient availability for microorganisms (Zhao and Zhu, 1997). The pH, temperature, the type and number of microbial species involved in degradation may influence the rate and extend of the process (Colwell and Walker, 1977). Apart from the presence of microorganisms capable of degrading crude oil compound the type, concentration and distance of pollutant, limit the biodegradation of the inherent compounds and other factors also involved in biodegradation. If the hydrocarbon is not available to the microorganisms and the appropriate enzymes, do not come in contact with the substrate in the proper way degradation will not occur (Atlas, 1981). Mainly enzyme activity, chemo taxis property and surfactants production of microorganisms response to the hydrocarbon degradation. Recently the soil bacterium Pseudomonas bacterium was reported to be attracted to the pollutant naphthalene and other chemicals. This expanded the range of organic compounds that are known to serve as bacterial chemo attractants to include aromatic hydrocarbons. However nothing is known about chemo taxis towards other common aromatic hydrocarbons such as toluene and benzene. Chemo taxis gives motile bacteria the advantage of the being able to locate compounds such as toluene that can support their growth (Parales et al., 2000). Biosurfactants are surface-active substances synthesis by living cells. They have the properties of reducing surface tension, stabilizing emulsion, promoting foaming and are generally non toxic and biodegradable, Interest in microbial surfactant has been steadily increasing in recent years due their diversity, environmentally, friendly nature, possibility of large-scale production, selectivity, performance under extreme condition and potential application in environmental protection (Rahman et al., 2002). Biosurfactants enhance the emulsification of hydrocarbons, have the potential to solublize hydrocarbon contaminants and increase their availability for microbial degradation. Biosurfactants producing microorganisms may play an important role in the accelerated bioremediation of hydrocarbon contaminated sites (Rosenberg and Ron, 1999). These compounds can also used in enhanced oil recovery and may be considered for their potential application in environmental protection. This research was aimed at determining the hydrocarbon degrading microorganisms present in the oil contaminated soil. The determined microorganisms (Pseudomonas aeruginosa and Pseudomonas fluorescens) were used to study the efficiency of the aromatic hydrocarbon (toluene) degradation on various concentrations. Simultaneously studied their chemo taxis properties and biosurfactant production of the above same microorganisms and also it was compared to MTCC culture of Pseudomonas stutzeri.
MATERIALS AND METHODS
Sample collection
The soil sample (100g) was collected from diesel contaminated site in a polythene bag for the isolation of oil degrading microorganisms. Samples were taken with shovels after removing 20 cm of the surface layer of the soil. Collected soil was stored aerobically at 4ºc for further analysis.
Isolation, identification and characterization of bacteria from oil contaminated soil enrichment technique
The hydrocarbon degrading microorganism was isolated by following the enrichment technique prescribed by Karpagam and Lalitha Kumari (1998). Identification was based on the criteria of Bergey’s Mannual of Determinative Bacteriology. Physico-Chemical Parameters Both the pH and moisture content of sample was analyzed by standard method and also the chemical parameters of carbon, phosphorous and potassium content of the sample was analyzed by standard methods. Hydrocarbon Estimation Hydrocarbon extraction After incubation, soil and hexane (1:20) were mixed for 5 minutes in a vortex and soil free hexane extracts was separated using a membrane filter and it was dried. The dried samples (dissolved hydrocarbon) were again dissolved HPLC methanol. After dissolving the sample in solvent, it was used for HPLC analysis. Hydrocarbon Determination The extracted hydrocarbon was analyzed by HPLC (C18 silica capillary column: 10AT VP Shimadzu company). Peak obtained at 5 minutes (retention time) has been considered as level of hydrocarbon degradation. Hence it was compared to the respective standard peak. Chemo Taxis Assay Agarose plug assays were carried out with slight modification. Plugs containing 0.5% of low melting temperature agarose in chemo taxis buffer. Chemo taxis Buffer (Potassium phosphate - 40mM Glycerol - 0.05%EDTA - 10mM). 10% (vol/vol) toluene and a few crystals of coomassie blue were used to provide contrast. A drop (20µl) of the molded agarose mixture was placed on a microscopic slide and the harvested cells flooded in to the microscopic slide to surround the agarose plug and incubate for 30minutes. After incubation, results were noted.
Bacterial Growth and Biosurfactant Production
A series of 500ml flask containing 200ml of sterile mineral salt medium with 1% glucose as a substrate were prepared, and the pH was maintained at 7.5. Each of the individual bacterial cultures was inoculated and the flasks were incubated at 30ºC in a shaker at 200rpm followed by the addition of 1% toluene after 24 hours. At every 24 hours intervals, biomass, biosurfactant production and emulsification activity were measured.
RESULTS
In this study, the bacterial strains were isolated from oil contaminated site in Sivakasi. From the oil contaminated soil, 10 bacterial strains were isolated. Among them, only two strains were used for further studies because they have the ability for survive that oil contaminated area.
Isolation and Characterization of Isolates
The cultural morphological and biochemical characterization of isolates were carried out and results were noted. Various biochemical and carbohydrates tests were carried out for isolated microorganisms and identified as Pseudomonas sp. Physico-Chemical Parameters The normal soil sample showed pH 7.9, but the oil contaminated soil sample showed low pH 6.7 than normal soil. The moisture content of the normal soil showed 0.908gms. But the oil contaminated soil showed low value (0.798gms) than normal soil (Table.1). The carbon content of the normal soil was found to be 16.8 (Kg/ha) but the carbon content of the oil contaminated soil was 22 (Kg/ha), because the oil contain increased content of carbon and hydrogen in their composition. In this study, the carbon, phosphorous and potassium content of the normal and oil contaminated soil was estimated. The results showed higher carbon content in the oil contaminated soil, because the oil contain large amount of hydrocarbons in their composition. During analysis of phosphorus estimation, the normal soil showed 17.9 (Kg/ha) of phosphorus content, but in the oil contaminated soil 11.2 (Kg/ha) of phosphorus content. During the estimation of presence of potassium in the normal soil, it was found to be 43.6 (Kg/ha). Whereas, the oil contaminated soil was found to be 30.2 (Kg/ha) of potassium. In the above study revealed that phosphorus and potassium content was rapidly metabolized by microorganisms amended with oil contaminated soil.
Degradation of Hydrocarbon (Toluene)
The degradation of hydrocarbon was analyzed by High Performance Liquid Chromatography (HPLC). Among the different strains of Pseudomonas sp. Pseudomonas fluorencens was found to be potassium in degrading the hydrocarbons (Table.2). When the concentration of toluene was increased, the rate of degradation was found to be decreased in the all experiments. Toluene and related aromatic hydrocarbons are highly toxic for living organisms because the preferential partitioning of these compounds in cell membrane disturbs the membrane structure, which leads to cell death. Hence the present investigation of toluene degradation was done, investigate certain the role of hydrocarbon which cause contamination to the environment. An oil spills in the environment leads to an adaptive process and if metabolically active hydrocarbon utilizing microorganisms could be added quickly, the long period before the indigenous population could respond would be reduced considerably (Chhatre et al., 1996).
Chemotaxis Assay
In the chemo taxis assay, a chemotactic response was observed in the form of a band of cells that accumulated in a ring surrounding, but not touching, toluene containing agarose plug. The Pseudomonas fluorescens was showed positive result to the chemo taxis assay within 30minutes and the Pseudomonas aeruginosa was showed positive result to the chemo taxis assay with in 45 minutes but Pseudomonas stutzeri was showed positive result to the chemo taxis assay after 45 minutes (within 1 hour). Besides, the roles of biosurfactant property in biodegradation were analyzed (Fig.1). Generally aromatic hydrocarbon compounds bind to soil components and are difficult to remove or degrade due to their limited availability to microorganisms.
Culture Biomass
During biosurfactant production, simultaneously the culture biomass was also observed and calculated. It was found out that total biomass of Pseudomonas fluorescens was increased from 1.021 to 1.20gms/ 100ml during 12 hours to 72 hours of incubation. Simultaneously the Pseudomonas aeruginosa showed 1.03grams/100ml during 12 hours of incubation and increased to 1.23 grams/100ml during 72 hours. Simultaneously growth of Pseudomonas stutzeri was found to be 1.01grams/100ml in 12 hours, increased to 1.21 grams/100ml during 72 hours of incubation. Emulsification Rate Emulsification rate was measured by height of the emulsion layer divided by total height of the reaction mixture and multiplying in to hundred. The rate was increased in Pseudomonas fluorescens 10 to 17.3%, Pseudomonas aeruginosa 8.4 to 16.6% and Pseudomonas stutzeri 7.5 to 16. 3% (Table.3).
Thin Layer Chromatography
After the production of biosurfactant was confirmed by the thin layer chromatography analysis and the Rf value (relative frond) value was calculated. The Rf value calculated from distance traveled by solute dividedly the distance traveled by the solvent on thin layer plates were calculated. The biosurfactant play an important role in degradation of hydrocarbons. In this study, the Pseudomonas sp. was found to be capable of producing surfactants exhibited emulsification activity on toluene. Emulsification activity is an indicator used extensively to enhanced identify and quantify biosurfactants produced by microbial culture of three Pseudomonas sp. Among them, three different species of Pseudomonas, Pseudomonas fluorescens revealed higher emulsification activity and also the amount of biosurfactant production simultaneously increased when compared to the Pseudomonas stutzeri and Pseudomonas aeruginosa. Crude surfactants were detected by Thin Layer Chromatographic technique (Fig. 2). Distance traveled by the solute Rf value = --------------------------------------- Distance traveled by the solvent
DISCUSSION
The oil spillage introduces non-organic and growth inhibiting chemicals present in the crude oil their toxicity to microorganisms and man is well known. The natural recovery of crude oil from polluted soils is slow (Ebuehi et al., 2005).Generally the natural degradation is show due to various reasons; they may be bioavailability, physical, chemical factors unsatisfactory. pH, oxygen requirement, moisture, redox potentsial, concentration of pollutant, recalcitrance (basic chemistry of pollution responsible), and xenobiotic nature of pollutant. Bioremediation ameliorates these factors through optimization of environmental conditions (Romas et al., 1995). In this study, the carbon, phosphorous and potassium content of the normal and oil contaminated soil was estimated. The results showed higher carbon content in the oil contaminated soil, because the oil contain large amount of hydrocarbons in their composition. Hence after oil spills, carbon content of the contaminated soil was increased than carbon level of normal soil. The microorganisms use the hydrocarbon as their carbon and energy source. The low amount of phosphorous and potassium content in the oil contaminated soil was found to be decreased than normal soil, because the available nutrients were rapidly assimilated by microbes, thus depleting the nutrient reserves in the contaminated soil. This coincides with the work of Vasudevan and Rajaraman, 2001. Where in they here described as increase in concentration leads to increase in microbial activity and role of biodegradation following adding the inorganic nutrients.
The biosurfactant play an important role in degradation of hydrocarbons. In this study, the Pseudomonas sp. was found to be capable of producing surfactants exhibited emulsification activity on toluene. Emulsification activity is an indicator used extensively to enhanced identify and quantify biosurfactants produced by microbial culture of three Pseudomonas sp. Among them, three different species of Pseudomonas, Pseudomonas fluorescens revealed higher emulsification activity and also the amount of biosurfactant production simultaneously increased when compared to the Pseudomonas stutzeri and Pseudomonas aeruginosa. Crude surfactants were detected by Thin Layer Chromatographic technique. Besides, the roles of biosurfactant property in biodegradation were analyzed. Generally aromatic hydrocarbon compounds bind to soil components and are difficult to remove or degrade due to their limited availability to microorganisms. This is in accordance with the work of Banat et al., 2000. According to him the biosurfactant (BS) can emulsify hydrocarbons, thus enhancing their water solubility, decreasing surface tension and increasing the displacement of oily substances from soil particles. In specific, among the three bacterial symbiasis such as Pseudomonas fluorescens, Pseudomonas aeruginosa and Pseudomonas stutzeri carried out for degradation studies, Pseudomonas fluorescens proved to be efficient in degradation of toluene, production of biosurfactant and chemotaxis property. Hence the present investigation entitled “Microbial Degradation of Hydrocarbons from Oil Contaminated Soil by using Pseudomonas sp” was studied.
CONCLUSION
Among the Various types of microorganism used in biodegradation process, pseudomonas Sp., in specific Pseudomonas aeruginosa, Pseudomonas fluorescence and Pseudomonas stutzeri play a vital role in degradation of hydrocarbons hence the present investigation, by using Pseudomonas sp., to degrade the toluene hydrocarbon. Pseudomonas sp., is recognized has one of the most important ecofriendly microorganisms in the world for clean-up technology by an adapting manner. The enzyme path way and their role in hydrocarbon degradation studies will be perform in future research.
Englishhttp://ijcrr.com/abstract.php?article_id=1999http://ijcrr.com/article_html.php?did=19991. Atlas, R.M. (1995) Efficacy of bioremediation chemical and risk based determinations. In Bioremediation: The Tokyo 94 Workshop. OECD Documents, Paris. France.
2. Atlas, R.M. (1981) Microbial Degradation of Pertroleum Hydrocarbon. An Environmental perspective. Microbial. reviews 45, 180 – 209.
3. Banat, I.M., Makkar, R.S. and Cameotra, C.S. (2000) Potential commercial application of microbial surfactance. Applied and Environmental Microbiology 58(5), 495-508.
4. Chhatre, S., Purohid, H.J., Shanker, R. and Khanna, P. (1996) Bacterial consortia for crude oil spill remediation. Water Sci.Technol 34, 187-193.
5. Colwell, R.R. and Walker, J.D. (1977) Ecological Aspects of Microbial Degradation of Petroleum in the Marine Environment CRC Critical Reviews in Microbiology 5(4), 423- 445.
6. Desai, J.G., and Banat, I.M. (1997) Microbial production of surfactants and their commercial potential. Microbiology and Molecular biology Reviews 61, 47- 65.
7. Ebuehi, O.A.T., Abibo, I.B., Shekwolo, P.D., Sigismund, K.I., Adoki, A and Okoro, I.C. (2005) Remediation of Crude Oil Contaminated Soil by enhanced Natural Attenuation Technique. J.Appl. Sci.Environ 9(1), 103- 106.
8. Isken, S. and De Bont, J.A.M. (1996) Active efflux of toluene in a solvent resistant bacterium. J. Bacteriol 178, 6056-58.
9. Karpagam, S. and Lalitha Kumari, D. (1998) Enumeration of hydrocarbon degrading bacteria in the petroleum polluted soils on Chennai city. Indian J. Env. Prot 18, 520.
10. Parales, R.E., Ditty, J. and Harwood, C.S. (2000) Toluene Degrading Bacteria Are Chemotactic towards the Environment Pollutant Benzene, Toluene and Trichloroethylene. Applied Environmental Microbiology 4098-04.
11. Rahman, K.S.M., Rahman, T.J., Lakshmanaperumalsamy, P. and Banat, L.M. (2002) Towards efficient crude oil degradation by a mixed bacterial consortium. Bioresource Technology 85, 257-261.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareBILATERAL FORAMEN ARCUALE IN ATLAS VERTEBRA: A RARE CASE
English8688S. D. DesaiEnglish Rekha HiremathEnglish Anand MugadlimathEnglishPonticuli in the atlas vertebra are bony spurs seen frequently from the anterior and posterior margins of the groove for the vertebral artery, which may convert the groove into a foramen.1 This foramen is known as foramen arcuale. The foramen arcuale is infrequently found and is potentially a clinically/surgically significant anatomical variation of the atlas. When present, the vertebral artery travels through this bony ring after exiting the transverse foramen of the atlas and prior to entering the cranium. We present a case report in which we noted during routine osteology practicals on atlas vertebrae for under graduates, foramen arcuale. The literature regarding these osteological structures is reviewed regarding their presence and potential clinical significance. The occurrence of bilateral fully developed posterior ponticuli resulting in encasement of the third part of the vertebral artery appears to be very rare, hence the case being reported.
Englishforamen arcuale, sulcus arteriae vertebralis, vertebrobasilar insufficiency.INTRODUCTION
A surgery pertaining to atlantoaxial complex has often posed a challenge to surgeons because of complex anatomy and biomechanics of this region of spine. The vertebral artery groove is located on the superior surface of the posterior arch of atlas at its junction with lateral mass lodging the third part of the vertebral artery, vertebral venous plexus and suboccipital nerve.1 The groove is also called as the sulcus arteriae vertebralis, sometimes, the distal part of the groove forms a full bone bridge to the upper edge of the Superior articular facet of the atlas, thus forming a foramen called as foramen-arcuale. The foramen arcuale is an anatomical variation that the neurosurgeon should consider when undertaking surgery near the posterior arch of atlas and also should be kept as a differential- diagnosis of patients presenting with symptoms of vertebrobasilar insufficiency. 2
Case Report
We report a case of bilateral foramen arcuale in human atlas vertebrae (Figure 2), found during routine osteology class for undergraduate teaching. Normal atlas is shown (Fig.1) for comparison with anomalous atlas vertebra. Complete fusion of the posterior ponticules was noted forming foramen for 3rd part of vertebral artery instead of groove (sulcus arteriae vertebralis).Vertebral artery represented by red colored wire in the figures below. In Fig.1 arrow represents sulcus arteriae vertebralis and in Fig.2 arrow represents posterior ponticle.
DISCUSSION
Many synonyms have been used by different authors from time to time some of them include‘foramen retroarticulare superior’ (Broacher, 1955), ‘canalis vertebralis’ (Wolff Heidegger, 1961), retroarticular vertebral artery ring (Lamberty and Zivanovic, 1973), ‘‘retroarticular canal’ (Mitchell, 1998) and ‘retrocondylar vertebral artery ring’ (Mitchell 1988).3 Ponticle formation stated by many authors is a regressive and morphological phenomenon; furthermore this anatomical variation is not related to an increase in degenerative changes synonymous with aging. These characteristics not only serve as anthropological data but may also help in identifying the impact of complete foramen arcuale on the signs and symptoms of vertebrobasilar insufficiency.4 The sulcus/groove for third part of vertebral artery on the dorsal aspect of the atlas that is completely covered by an abnormal ossification is termed the ponticulus posticus. The resulting foramen contains the vertebral artery referred as foramen arcuale. The foramen arcuale, an osseous variant of the atlas should be considered during lateral mass screw fixation.5 Complete ossification of ligamentous free margin of posterior atlanto-occipital membrane can lead to complete foramen arcuale. Clinical complaints such as vertigo, neck pain of discopathy are reported. Foramen arcuale is associated with Barre-Lieou syndrome, which represents symptoms of headache, retro-orbital pain, vasomotor disturbance of the face and recurrent disturbances of vision, swallowing and phonation due to alteration of blood flow within the vertebral arteries and an associated disturbance of periarterial nerve plexus.6 Foramen arcuale leads to symptomatic entrapment, additional compression of the vertebral artery by a lateral ponticle when present could very likely result in stenosis of the vertebral artery .7 In a study Limousin reported good results in patients with this syndrome and an identified foramen arcuale in which the foramen was fractured and a periarterial sympathectomy was performed8 . In another report Li et al. placed the foramen arcuale in the differential diagnosis for vertigo and found satisfactory results in eleven patients that underwent decompression of the vertebral artery in foramen arculae.9 Likewise, Sun found cessation of vertigo in 69 patients undergoing decompression and sympathectomy of the vertebral artery within a foramen arcuale.10 Cushing et al. noted foramen arcuale in 8 of 11 patients with vertebral artery dissection and occlusion.11 From the above discussion it is evident that foramen arcuale is a clinically significant anomaly of atlas vertebra which can present with varied clinical features and syndromes, and hence the case being reported for publication.
CONCLUSIONS
The knowledge about this present anatomical variation will be of immense help to Neurologists, neurosurgeons and radiologists when dealing with the patients complaining of symptoms of vertebrobasilar insufficiency like headache, vertigo, shoulder and arm pain.
ACKNOWLEDGEMENT
Authors acknowledge the 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=2000http://ijcrr.com/article_html.php?did=20001. Cakmak O, Gurdal E, Ekinci G, Yildiz E, Cavdar S. Arcuate foramen and its clinical significance. Saudi Med J 2005; 26: 1409- 1413.
2. Tubbs RS, Johnson PC, Shoja MM, Loukas M. Foramen arcuale: anatomical study and review of the literature. J Neurosurg Spine. 2007; 6: 31–34.
3. Hasan M, Shukla S, Siddiqui MS, Singh D. Posterolateral tunnels and ponticuli in human atlas vertebrae. J Anat. 2001; 199: 339–43.
4. Unur E, Erdogan N, Ulger H, Ekinci N, Ozturk O. Radiographic incidence of complete arcuate foramen in Turkish population. Erciyes Medical Journal. 2004; 26: 50–54.
5. Young JP, Young PH, Ackermann MJ, Anderson PA, Riew KD. The ponticulus posticus: implications for screw insertion into the first cervical lateral mass. J Bone Joint Surg Am. 2005; 87: 2495–98.
6. Limousin CA. Foramen arcuale and syndrome of Barre-Lieou. Its surgical treatment. Int Orthop. 1980; 4: 19–23.
7. Tubbs S, Shoja MM, Shokouhi G, Farahani RM, Loukas M, Oakes JW. Simultaneous lateral and posterior ponticles resulting in the formation of a vertebral artery tunnel of the atlas: Case report and review of the literature. Folia Neuropathol. 2007; 45: 43–46.
8. Limousin CA. Foramen arcuale and syndrome of Barre-Lieou. Int Orthop 1980; 4: 19-23.
9. Li S, Li W, Sun J. Operative treatment for cervical vertigo caused by foramen arcuale. Zhonghua Wai Ke Za Zhi 1995; 33: 137-139.
10. Sun JY. Foramen arcuale and vertigo. Zhonghua Wai Ke Za Zhi 1990; 28: 592-594.
11. Cushing KE, Ramesh V, Gardner-Medwin D, Todd NV, Gholkar A, Baxter P, Griffiths PD. Tethering of the vertebral artery in the congenital arcuate foramen of the atlas vertebra: a possible cause of vertebral artery dissection in children. Dev Med Child Neurol 2001; 43: 491-496.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareSYMPATHETIC REACTIVITY TO COLD PRESSOR TEST IN MEDICAL STUDENTS OF HYPERTENSIVE AND NORMOTENSIVE PARENTS
English8993Rinku GargEnglish Yogesh TripathiEnglish Varun MalhotraEnglishBackground: Essential hypertension shows familial predisposition. It is detectable at an early age in descendants of hypertensive parents following sympathetic stimulation. Aims and objectives: The present study was designed to examine the response in the blood pressure to the cold pressor test in medical students of hypertensive parents and to compare it with age-matched controls of medical students of normotensive parents. Material and Methods: Cold pressor test was performed in the study and control groups. First basal blood pressure was recorded and afterwards subjects were asked to dip left arm in the cold water (temperature at 2-40 C) for 2 minutes and blood pressure was recorded from the right arm in sitting position. Blood pressure was again recorded 5 minutes after hand was taken out of the cold water. Results and conclusions: Results showed that basal systolic and diastolic blood pressure were higher (p Englishhypertension, cold pressor test, baseline blood pressureINTRODUCTION
Blood pressure is defined as the lateral pressure exerted by the column of flowing blood on the blood vessel wall. Hypertension is the increase in blood pressure ≥140/90 mm Hg measured on three separate occasions1. In about 88% of the patients with hypertension, cause of hypertension is unknown and they are said to have essential hypertension. Essential hypertension is polygenic in nature (2).Genetic factors as well as environmental factors play an important role in the development of primary hypertension3-6. High blood pressure before the age of 55 years occurs 3.8 times more in individuals with a family history of hypertension6 . Sympathetic system has been implicated to be overactive in these individuals7-11.Individuals developing hypertension at a later age might have an altered physiology at a very young age12.Blood pressure regulating mechanisms have been shown to be hyperactive in these individuals13.There are different sets of genes that regulate blood pressure during rest and during cold pressor test and are independent of each othe5 . Main circulatory alteration in essential hypertension might be due to increased vascular peripheral resistance or a raised cardiac output15.People at high risk for high blood pressure may have an exaggerated stress-induced cardiovascular response at a younger age13. Cold pressor test is a satisfactory method in studying the reactivity of blood pressure14. Cardiovascular reactivity to stress has pathophysiological role in hypertension12. . The aim of the present study was to examine the blood pressure response following sympathetic stimulation in the form of cold pressor test in the medical students of hypertensive parents and compare it with the age-matched controls of normotensive parents. MATERIAL AND METHODS The present study was a cross-sectional study, conducted in Santosh Medical College, Ghaziabad .Ethical approval was taken from the research committee of the Institution. Fifty medical students in the age group of 17-24 years with family history of hypertension (either single parent or both parents) were recruited for the study and the results were compared with fifty age-matched medical students with no familial history of hypertension. Informed consent was taken from all the medical students. Medical students with H/o any chronic illness, on any medication, smokers were excluded from the study. Subjects were divided into 2 groups Group I-control group with no family h/o hypertension Group II-study group with family h/o hypertension Procedure of the cold pressor test (CPT) was explained to all the students participating in the study. Before the test, subjects were allowed to rest for 10 minutes in a quiet room to reduce the anxiety. Basal blood pressure of all the subjects was measured by auscultatory method with the help of mercury sphygmomanometer (DIAMOND).First Kortkoff sound indicated systolic blood pressure (SBP) and fifth Kortkoff sound indicated diastolic blood pressure(DBP). Mean blood pressure (MBP) was calculated as diastolic blood pressure + 1/3rd of Pulse Pressure. Cold pressor test was done in both the study group and control groups. After recording basal blood pressure, subjects were asked to dip left arm in the cold water (temp at 2- 4 0 C) for 2 minutes and blood pressure was recorded from the right arm. Blood pressure was again recorded 5 minutes after hand was taken out from the cold water.Results were analysed by ANOVA with SPSS version 17.0 using unpaired‘t’ test
RESULTS
Data presented in Table 1 shows that there was significant increase in the basal SBP,DBP &MBP in the individuals as compared to group I and values were statistically significant(pEnglishhttp://ijcrr.com/abstract.php?article_id=2001http://ijcrr.com/article_html.php?did=20011. World Health Organisation Expert Committee:Hypertension Control Technical Report Series 862 Geneva,WHO 1996.
2. Ganong WF.Cardiovascular homeostasis in health and disease.In:Ganong WF,ed.Review of medical physiology.22nd ed.Newyork :McGraw Hill:2005,p,631-646.
3. Mcllhany ML,Shaffer JW,Hines EA Jr: The heritability of blood pressure :an investigation of 200 pairs of twins using the cold pressor test.John Hopkins Med J 1975;136(2):57-64.
4. Thacker EA:A comparative study of normal and abnormal blood pressure among university students,including the cold-pressor test.American Heart Journal 1940;20(1):89-97.
5. Busjahn A,Faulhaber HD,Viken RJ,Rose RJ,Luft FC:Genetic influences on blood pressure with the cold-pressor test:a twin study.J Hypertens 1996;14(10):1195-9.
6. Williams RR,Hunt SC,Hasstedt SJ,Hopkins PN,Wu LL,Berry TD,Stults BM,Barlow GK,Schumacher MC,Lifton RP: Are there any interactions and relations between genetic and environmental factors predisposing to high blood pressure. Hypertension 1991;18(3):129- 37.
7. Mancia G,Di Rienzo M,Giannattasio C,Parati G,Grassi G:Early and late sympathetic activation in hypertension.Scand Cacrdiovasc J Suppl 1998;47:9-14
8. Grassi,Guido:Role of sympathetic nervous system in human hypertension.Journal of Hypertension 1998;16(12):1979-1987.
9. Quattro VD,Feng M.The sympathetic nervous system:the muse of primary hypertension.Journal of Human Hypertension 2002;16(1):s64-s69.
10. Khaliq F,Gupta K,Singh P. Autonomic reactivity to cold pressor test in prehypertensive and hypertensive medical students.Indian J Physiol Pharmacol 2011;55(3):246-252.
11. Parmanik T,Regmi P,Adhikari P,Roychowdhury P.Cold pressor test as a predicto.r of hypertension.J The Univ Heart Ctr 2009; 3:177-180
12. Mathews KA,Woodall KL,Allen MT.Cardiovascular reactivity to stress predicts future blood pressure status.Hypertension 1993;22:479-485.
13. Menkes MS,Matthews KA,Krantz DS,Lundberg U,Mead LA,Qaqish B,Liang KY,Thomas CB,Pearson TA:Cardiovascular reactivity to the cold pressor test as a predictor of hypertension.Hypertension 1989;14(5):524- 30.
14. Edgar A,Hines Jr:The significance of vascular hypertension as measured by the cold- pressor test.American Heart Journal 1940;19:408-416.
15. Hooft IMV,Grobbee DE, Waal-Manning HJ,Hofman A.Hemodynamic characteristics of the early phase of primary hypertension.The Dutch Hypertension and offspring study.Circulation 1993;87:1100-1106.
16. Walia V,Ahuja J,Kaur P.Cardiovascular reactivity to cold pressor test in offspring ofnormotensive and hypertensive parents.The Indian Journal of Medical Research 1994;99:38-41.
17. Douglas C, George DS,Gonneke W,David S,Peter M S,JohnEJG,Peter CE.Blood pressure reactions to the cold pressor test and the prediction of ischaemic heart disease:data from the carephilly study.J Epidemiol Community Health 1998;52:528-529.
18. Verma V,Singh SK,Ghosh S:Identification of susceptibility to hypetension by the cold pressor test.Indian J Physiol Pharmacol 2005;49(1):119-120. )
19. Gupta AK.Influence of family history of morbid cardiovascular events on blood pressure of school children.Indian Paediatr 1991;28(2):131-9.
20. Thomas J. Meharry Cahort study.J Natl Medl Assoc.82(6):409-412.
21. Nageswari KS,Sharma R,Kohli DR.Assessment of respiratory and sympathetic cardiovascular parameters in obese school children.Indian J Physiol Pharmacol 2007;51(3):235-243.
22. Rajashekhar RK,Niveditha Y,Ghosh S.Blood pressure response to cold pressor test in siblings of hypertensives.Indian J Physiol Pharmacol 2003;47(4)453-458.
23. Velasco M,Gomez J,Blanco M,Rodriguez I.The cold pressor test:pharmacological and therapeutic aspects.American Journal of Threaputics 1997;4(1):34-38.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareSTUDY OF SOCIODEMOGRAPHIC PROFILE OF CONTRACEPTIVE USERS IN AN URBAN SLUM OF BIJAPUR
English9499Santosh M BiradarEnglish Angadi M MEnglishBackground: The fruits of economic growth cannot felt unless population explosion is prevented. The contraceptive usage in community depends on various factors like socioeconomic class, religion, education, parity, preference for male children etc. The study was done to know the sociodemographic profile of contraceptive users Objective: To assess the prevalence of contraceptive usage and to find out reasons for non acceptance. Methodology: community based cross-sectional study of the married women residing in an urban slum of Bijapur. Results: couple protection rate was 65.3%. Tubectomy (56.6%) was most common method of contraception. Contraceptive usage was more in families were women was from upper class, nuclear family, higher age group, high parity and had sons. These associations were found to be statistically significant. 34.7 % women were non acceptors of contraception. The most common reason for non acceptance was pregnancy and lactation (34.6%) and intending for more children (24.1%). Other important reasons were fertility related reasons, Opposition to use, Lack of awareness, and Fear of side effects. Conclusion: Contraceptive usage in community can be increased by improving IEC activities and quality of family planning services.
Englishsociodemographic profile, contraceptives, family planning.INTRODUCTION
An Expert Committee (1977) of the WHO defined family planning as "a way of thinking and living that is adopted voluntarily, upon the basis of knowledge, attitudes and responsible decisions by individuals and couples, in order to promote the health and welfare of the family group and thus contribute effectively to the social development of a country" 1 .The population growth rate in India is alarming and detrimental to development of country. The fruits of economic growth cannot felt unless population explosion is prevented. According to the census 2001, the Indian population was 1024 million and is expected to overtake China and become the most populous country in the world by 2045.2 India was the first country in the world to formulate the National Family Planning Programme in year 1952 with objective of “reducing the birth rate of the extent to stabilize the population at a level consistent with the requirement of National Economy” 3 . The contraceptive usage in community depends on various factors like socioeconomic class, religion, education, parity, preference for male children etc. The study was done to know the sociodemographic profile of contraceptive users and also to find out reasons for non acceptance.
MATERIALS AND METHODS
This is a community based cross-sectional study. The study was conducted on married women (15yr – 49yr) residing in an urban slum of the field practice area of Shri B M Patil Medical College. Sample size was calculated using the formula4 Z= 4PQ / D2 as 258 ( NHFS-3 5 , prevalence (P) of Family Planning practices in Karnataka for urban population was 60.8 %, at 5 % significance, with 10 % allowable error). The total sample size was rounded off to 300. The participants were interviewed using a predesigned, structured questionnaire and data was analysed by statistical tests like proportions and chi-square.
RESULTS AND DISCUSSION
It was observed from the study that majority women were Hindus (71.3 %) and Muslims were 28.7%.Most women were in the age group of 20-29yrs (29.6%) and from joint family (59%). As the study was conducted in slum, majority women (62.3%) were from lower socioeconomic strata (upper Lower and Lower class of Modified Prasad Classification).
The couple protection rate in study area was 65.3% with 196 contraceptive users. The 34.7% women were not using any form of modern contraception. The contraceptive prevalence in study area (65.3%) was more than that observed by NFHS-3 for urban population in Karnataka (60.8%). Tubectomy (56.6%) was most common method of contraception. This was followed by methods like OCP (18.4%), IUD (16.3%) and Condom (8.7%). None of subject's partner had undergone Vasectomy as a method of contraception. A study in rural community of Maharashtra 6 observed the prevalence of contraceptive usage of 66.7 % with tubectomy as most common method of contraception. Anju Puri et al7 reported 58.3% of contraceptive users had tubectomy and vasectomy was not accepted by any of subject's partner. These studies indicate male dominant nature of society with family planning as a female domain. Thus there is need to encourage male participation (condoms, vasectomy) and improve couple protection rate.
It was observed that 34.6% of women were either Pregnant or Lactating and hence not using any contraception. Intenders for children accounted for 24.1%.The other reasons for non acceptance were Opposition to use (14.4%), Fertility related reasons (11.5%), Lack of awareness (9.6%), Fear of side effects (5.8%). Khokhar A8 et al reported in their study reasons for non acceptance as anti religion, opposition from family members, ignorance about use and no knowledge about the source. These studies indicate that many subjects had given the reason for not using contraception as lack of information, worry of side effects and opposition to use. These couples need to be properly counseled to reduce the myths and fear about contraceptives and improve acceptance rates.
The age of women had an influence on acceptance of contraception. The acceptance rate increased with increase in age of women. This association was highly significant (p 0.05). DHS reports8 from Bangladesh, Nigeria, Pakistan, Senegal and study by Padma Mohanan10 et al have shown Muslims were less likely to be ever users. The non acceptors are more in Muslims may be due to religious objections.
Acceptors of contraceptives were more from the nuclear families (73.2%) compared to joint families (59.9%) and this association was statistically significant (PEnglishhttp://ijcrr.com/abstract.php?article_id=2002http://ijcrr.com/article_html.php?did=20021 WHO (1971). Techn. Rep. Ser., No. 483
2 Park K. Textbook of Preventive and Social Medicine 19th edition published by Banarsidas Bhanot Jabalpur 2007. Page No 389-412
3 National Population Policy-2000, Government of India
4 Kishore J. National Health Programmes of India; National policies and legislations related to health. Century publications New Delhi 7th edition, page no 93
5 National Family Health Survey, India. Available at http://www.nfhsindia.org
6 Balaiah, M. Ghule, Naik D D, Parida R C and Hazari K T. Fertility attitudes and family planning practices of men in rural community of Maharashtra. The Journal of Family Welfare, 2001; April; Vol. 47, No.1, Page 56-37
7 Anju Puri, Suneela Garg, Malti Mehra. Assessment of unmet need for contraception in an urban slum of Delhi. Indian Journal of Community Medicine, (2004-07 - 2004-09); Vol. 29, No. 3
8 Khokhar A, Gulati N. A study of never users of contraception from an urban slum of Delhi. Indian Journal of Community Medicine, (2000-01 - 2000-03); Vol. 25, No. 1
9 Sumedha M. Joshi, Shirish B.Patil. Knowledge and practice of family planning in an urban slum of Mumbai, Indian Journal of Preventive and Social Medicine;2007; Vol. 38, No 1 and 2
10 Padma Mohanan, Asha Kamath, B.S. Sajjan. Fertility pattern and family planning practices in a rural area in Dakshina Kannada. Indian Journal of Community Medicine 2003; Jan.-Mar; Vol. Xxviil, No.1
11 Bratati Banerjee. Socioeconomic and cultural determinants of acceptance of permanent methods of contraception. The Journal of Family Welfare, 2004; June; Vol 50 No 1, Page No 54-60
12 Rama Ram, Ghosh M.N, Salil Bhattacharya, Anima Haldar, Chitra Chatterjee, Narendranath Naskar. Study of unmet need for family planning among married women of reproductive age attending immunization clinic in a medical college of Calcutta. Indian Journal of Community Medicine. (2000-01 - 2000-03); Vol. 25, No. 1
13 Kansal A, Chandra R, Kandpal S.D, Negi KS. Epidemiological correlates of contraceptive prevalence in rural population of Dehradun district. Indian Journal of Community Medicine, 2005; April-June; Vol. 30, No. 2, Page 60-62
14 Sharma AK, Grover V, Agrawal OP, Dubey KK, Sharma S. Pattern of contraceptive use by residents of a village in South Delhi. India Journal of Public Health 1997; 41 (3): 75-78.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcarePREVALENCE OF ANAEMIA IN SCHOOL CHILDREN OF BIJAPUR, KARNATAKA
English100108Shamshad Begum .A.LoniEnglish Irshad AhamedEnglish Lizzy SunnyEnglishObjectives: Prevalence of anemia is high in India .Anemia is a serious health problem in Indian school children. Material and methods: Children of age group 6-15 years of Secab institution of Bijapur district of Karnataka were included in our study .Hemoglobin was determined by Salhi’s method taking WHO standard .Result. Of the 1033 students included in the study, 594 were boys and 439 were girls. Overall prevalence of anemia was 67.1% with higher prevalence in girls (79.7%) as compared to boys (57.7%). The difference in prevalence was higher in the age-group >12 years (boys = 48.5%, girls = 82.5%) as compared toEnglishAnemia, Salhi’s method, HemoglobinINTRODUCTION
Anemia is a serious health problem in Indian school children. The nutritional status of children is a good indicator of the health status of a community1.According to Dreyfuss et al (2000)2 two billion children are affected with iron deficiency anemia worldwide. There is now need for the development of clear policy guidelines based on these simple and integrated intervention3 . Anemia is estimated to effect one half (50%) of the school age children in developing countries4 . Iron deficiency anemia affects a substantial portion of the world population pocking severe health problem to the people suffering these condition5 . Iron deficiency is probably the only nutrient deficiency of significant prevalence in virtually all developed countries 6 . Recent studies on the nutritional status of Indian children in this age group have showed prevalence ranging from 10- 60 % 7, 8, 9,10,11,12.Adolescence is defined by WHO as the period of life between 10- 19 years (WHO 1996)13 .Causes of anemia are manifold ranging from nutritional deficiencies which includes iron deficiency, which is the world’s largest nutritional problem (WHO) B12 and folate deficiency14. Dietary intake of haem iron (animal origin) is negligible in developing countries. Ascorbic acid, presence of food items of animal origin and low PH enhance while milk, coffee and tea inhibit the iron absorption. Likewise, deficiency of Vitamin A limits the body ability to use stored iron, resulting in an apparent iron deficiency because Hb levels are low even though the body stores are normal15.Daily iron requirement of children aged 6-11 years is 40ug/kg/day.14 . Many south East Asian countries including India have started iron and folic acid supplementation in target population .Indonesia has covered 60% of its target population via iron and folic acid supplementatary16 . We Indian still have not quantified anemia in our school age population .Hence our study is an attempt to estimate the magnitude of this problem of anemia in apparently healthy school children of age group 6-16 years of both sexes. The united Nation’s subcommittee on Nutrition held in Oslo in 1998 concluded that more data on health and nutrition of school age children are needed to assess their scale of problems 17 . However, recent studies have shown that the prevalence of malnutrition and anemia is high in these age group18 . The most commonly used indicator of iron deficiency anemia is Hemoglobin. But it is true that not all anemias are due to iron deficiency and not all iron deficiency will reflect in anemia
Aim of study
The present study was conducted with the object of assessing the nutritional status of children of age group 6-15 years attending school in Bijapur. And to compare Hematological Measurements with the WHO standards for that age and sex groups.
MATERIALS AND METHODS
An observational descriptive cross sectional study was conducted in apparently healthy children of age group 6-15 years in Secab Intuitions of Bijapur.
Measurements
Hemoglobin was determined by Salhi’s method. WHO standard values were considered to compare the value for those age groups and sexes. Salhi’s method of hemoglobin estimation has been used since long20 . Inspite of the availability of newer techniques which give more reliable and accurate results, this method is still in vogue 21. In our study children having Hb level 11.5 were classified as having sever, moderate, mild and no anemia respectively22 (WHO 2001). Statistical analysis – was done by statistical package for SPSS software .Mean and SD of mean was computed for each group and sex.
RESULTS
The study was carried out in apparently healthy school children of age group 6-16 years attending the schools of Secab institution of Bijapur district of South India. Relevant history was taken and completes physical examination done. Of the 1033 students included in the study, 594 were boys and 439 were girls. Overall prevalence of anemia was 67.1% with higher prevalence in girls (79.7%) as compared to boys (57.7%). The difference in prevalence between boys and girls were higher in the age-group above 12 years (boys = 48.5%, girls = 82.5%) as compared to 12 years or below 12 years (boys = 65.6%, girls = 78.0%). Further stratification of the age into smaller groups showed a highly significant difference in the mean hemoglobin levels between boys and girls in the age-group 6 to 8 years (boys = p=0.002). Among older children, the difference in the mean hemoglobin levels between boys and girls were highly significant for 13, 14, &15 years (p=0.001), In the age group of 6-15 years our study showed the prevalence of Anemia of the total children screened (N=1033), 67.1% had Hb level below the normal (12 g dl) values, indicating anemia, out of which 57.7% were boys and 79.73% were girls.
In 57.75 of anemic boys 41.07% had mild and 16.66% had moderate anemia .In girls of 79.33%, 53.07% were mild and 26.65% were moderate anemia .In age group of < 12 years we could find 30.69% mild and 25.1% moderate . In girls 12 years the mild prevalence of anemia in boys was 38.21 % and moderate 12.19 % In girls it was 51.68% mild and 27.3% moderate out of which none had severe anemia... Table 1 of our study shows the number of school going children selected. Of 1033 children 594 boys (57.7%) and 439 girls (42.5%) of age group 6-15 years. Among 6-15 years age groups children selected highest number of children was in the age group of 10 years and 12-13 years. Table 2 of our study shows the overall % of anemia in the school children. Higher Prevalence of anemia was found in girls (79.7%) as compared to boys (48.5%).Among the girls of 6-15years high prevalence of anemia was found in age group of above 12 years. When compared to below 12 years group (78%). Among the e boys higher prevalence was seen in 12 years (48.5%). The mean hemoglobin values according to the both age in both sexes are shown in table 3: The mean hemoglobin values of boys and girls aged between 13 and 15 years were highly significant The mean hemoglobin values of boys and girls at age of 6,12 were not significant . The overall mean hemoglobin values between the boys and girls were highly significant. Table 3 depict the age wise mean_+SD of Hb with P value of 12 years when compared with boys of same age 48.5%.Similarly in girls Englishhttp://ijcrr.com/abstract.php?article_id=2003http://ijcrr.com/article_html.php?did=20031. Ujjwal Chakraborty,Sutanu Dutta,Chowdhury, Goutam Dutta, Tusharkanti Gosh.A Comparative study of Physical Growth and Nutritional Status in Santal Children of Ghatsila and Bolpur.Tribes and Tribals 2008,sp:(2):79-86.
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40. Zimmermann M.B., Zeder C, Chaouki N, Saad A, Torresani T, Hurrell RF.Dual fortification of Salt with iodine and microencapsulated iron: A randomized, double- blind, controlled trial in Moroccan school children. Am. J. Clin. Nutr2003 Feb. 77(2):425-432.
41. Ruchika Handa, Faizan Ahamad, Kavindra Kumar Kesari , Ranu Prasad. Assessment of Nutritional Status of 7-10 Years School Going Children Of Allahabad District: A Review: Middle East Journal of Scientific Research 2008;3(3):109-115.
42. Arlappa N, Balakrishna N, Laxmaiah A, Brahmam GNV. Prevalence of anaemia among rural pre-school children of West Bengal, India. Ann Hum Biol. 2010April; 37(2):231–242.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareRELATIONSHIP OF LOWER LIMB SPASTICITY, STRENGTH AND GROSS MOTOR FUNCTION IN CHILDREN WITH SPASTIC DIPLEGIA: A CROSS-SECTIONAL STUDY
English109122Kamat PA English Ganesan SEnglish Jaya Shanker TedlaEnglishPurpose: To establish the relationship between spasticity and strength in the lower limb muscles and gross motor function of children with spastic diplegia. Method: This cross-sectional study included thirty children (21 males, 9 females) with spastic diplegia of GMFCS-EandR levels I to V between the age of 5 to 15 years. Spasticity, strength and gross motor function were assessed using tardieu scale, hand-held dynamometer and Gross Motor Function Measure respectively. Results: The correlation between spasticity, strength and gross motor function was analyzed using the spearman’s correlation. A negative correlation was found between spasticity of hip flexors, hip adductors and knee extensors with GMFM. No correlation was found between spasticity and strength. A positive correlation was found between strength of hip abductor, knee flexors, dorsiflexors and plantarflexors with GMFM. Conclusion: Aggregate spasticity of muscles around the hip, knee and ankle joint showed a negative correlation with GMFM, whereas aggregate strength of muscles around all the 3 joints showed a positive correlation with GMFM. Hip flexor was the only muscle that showed a negative correlation between spasticity and strength.
EnglishINTRODUCTION
Cerebral palsy describes a group of disorders of the development of movement and posture, causing activity limitations that are attributed to non-progressive disturbances that occurred in the developing fetal or infant brain. The motor disorders of cerebral palsy are often accompanied by disturbances of sensation, cognition, communication, perception, behaviour or by a seizure disorder.1 Cerebral palsy (CP) has been generally classified on the basis of motor abnormalities as spastic, dyskinetic and ataxic.1 Based on the topographic distribution of impairments, spastic CP is divided into monoplegia, hemiplegia, diplegia, triplegia and quadriplegia.2 Spastic diplegia is the most common form of spastic cerebral palsy, occurring in 21.9 % of cerebral palsy children.2, 3 Children with spastic diplegia have many neurological deficits that interfere with motor function and daily activities. These impairments include neuromuscular and musculoskeletal problems such as spasticity, weakness, loss of selective motor control, incoordination and contractures.4 In children with spastic diplegia, lower limbs are more severely affected than upper limbs.2 Lower extremities exhibit abnormal timing of muscle contraction, poor ability to terminate muscle activity, and also show limited functional synergies and poor grading of co-contractions.5 Spasticity has been viewed as a major primary impairment of body function in spastic diplegics. Despite the lack of consensus on the role of spasticity in motor abilities, alleviations of spasticity remain the primary focus in the clinical management of spastic diplegics. Spasticity might produce contractures due to a marked loss of sarcomeres in the muscle, increase stiffness in the spastic muscle and changes connective tissues within the muscle. 4 Strength is an important aspect of normal motor control.6 Research finding indicate that children with spastic diplegia are indeed weak and that strength is directly related to motor function and therefore this weakness can be associated with difficulties in performing everyday functional activities.7 Gross motor development in children is commonly described as the acquisition of motor milestones such as unsupported sitting, crawling and walking. Children and infants with spastic diplegia are at risk for gross motor developmental disabilities. There is evidence that severity of the disorder is related to development of gross motor function. The primary goal of therapeutic intervention for children with spastic diplegia is to enhance the child’s ability to perform activities in the context of daily life. Relationship between motor impairments and functional activity has a significant impact on clinical practice.4, 8 Limited literature is available about the relationship of strength, spasticity, and gross motor function in children with spastic diplegia. The three variables of strength, spasticity and function have been correlated in various combinations at various times, but a general consensus regarding the relationship of the three has not been reached. The results of published literature shows high level of variation with some studies suggesting a significant relation between the three variables and many establishing insignificant or no relation.4,9-12 A study was done on children with spastic diplegia of GMFCS levels I to III to determine the relation between spasticity, strength, gross motor function and the functional measures of gait. They used Isokinetic dynamometer to measure spasticity and strength of selected ankle, knee and hip muscles. Gait analysis was done to evaluate linear and kinematic variables. They had also recorded gross motor function using GMFM 66. They concluded that spasticity did not account for a substantial amount of variance in gait and gross motor function. There was a moderate to high correlation between strength, gait linear data and gross motor function.10 Studies done on the relation between strength, spasticity, and gross motor function have used Isokinetic dynamometer to record strength or spasticity on children of GMFCS levels I to III. Most of the studies were done by either taking a single joint or all 3 joints of the lower limbor aggregate spasticity and aggregate strength.10, 11, 13 Isokinetic dynamometers are expensive instruments, requiring specific training for data collection and recording, which is usually provided by the manufacturers. This makes it inaccessible to many pediatric physiotherapy units. There are many cost effective, easy to perform reliable tools available for recording spasticity, strength and gross motor function like Tardieu, hand held dynamometer and GMFM-88 respectively.In children with spastic diplegia of GMFCS-EandR levels I to V, evidences regarding the relationship of spasticity, strength and gross motor functions are largely unknown. Studies regarding comparison of individual joint involvement with gross motor function are also limited. Aim of the current study was to establish the relationship between spasticity and strength in the lower limb muscles and gross motor function of the children with spastic diplegia of GMFCS-EandR levels I to V by using Tardieu scale, hand held dynamometer and GMFM-88 respectively. Objective of the study is to evaluate if individual joint involvement can have an effect on gross motor function of the children with spastic diplegia.
METHODOLOGY
This was a cross-sectional study using a convenient sampling method included 30 (21males and 9 females) children with spastic diplegia between the age of 5 to 15 years of GMFCS- EandR level I –V. These children were graded on Modified Child Mini Mental State Examination.19 Only those who had a minimum score of >24 for 5years, >28 for 6-8years, >30 for 9-11 years and >35 for 12-15 years were included. Those children who had an history of corrective surgeries, Botox / phenol injections within the past 6 months, any systemic or musculoskeletal disorders interfering with performance of child, children who are taking muscle relaxants (baclofen, dantrolene), or those who had fixed flexion contracture of hip and knee >25° and fixed ankle plantarflexion contracture of > 10° were excluded. The materials used were GMFCS-EandR, GMFM-88 scale, Baseline pushpull digital TM dynamometer, Tardieu scale, Omega Universal Goniometer.
Procedure
Approval from the scientific committee and time bound research ethics committee was obtained prior to commencement of the study. All parents of the children with spastic diplegia signed the consent form. Children aged 8 years and above signed the assent form. Children of both genders diagnosed by pediatrician as spastic diplegics were screened by the tester for inclusion and exclusion criteria. Children who fulfilled the inclusion criteria wereclassified on the GMFCS-EandR levels I toV.16, 23 Spasticity, strength and gross motor function was assessed using the Tardieu scale, hand held dynamometer and GMFM-88 respectively. In children with spastic diplegia the spasticity and strength of hip flexors, extensors, adductors and abductors, knee flexors, and extensors, ankle dorsiflexors and plantarflexors of both lower limbs were tested using Tardieu scale20,21,25 and hand held dynamometer14,26,32 respectively. Spasticity was assessed in one attempt only. The hand-held dynamometer was used to quantify isometric strength of the major lower extremity muscle groups bilaterally. Children were positioned in the respective test positions (Appendix-1). The proximal joint segment was stabilized. The plate of the dynamometer was then placed distally on the distal joint segment. The piston of the dynamometer was held perpendicular to the limb segment in the direction of the movement to be performed. The subject was then asked to produce a maximal isometric contraction of the muscle group, tested by pushing against the plate of the dynamometer and holding in position for duration of 1 second to allow the participant to adjust and recruit the maximum number of muscle fibers. Three attempts were recorded. 1st attempt was to familiarize the child with the test procedure and the final reading was obtained from the best out of the 2nd and 3rd readings. The Gross Motor Function Measure (GMFM)was administered to measure gross motor function quantitatively. The GMFM consists of 88 items grouped into 5 dimensions: (1) lying and rolling (2) sitting, (3) crawling and kneeling (4) standing and (5) walking, running, and jumping.17
Data Analysis
The values of spasticity and strength of hip flexors, extensors, adductors and abductors, knee flexors and extensors, ankle dorsiflexors and plantarflexors between right and left side were compared by using Mann-Whitney U test. The data of spasticity and strength of hip flexors, extensors, adductors and abductors, knee flexors and extensors, ankle dorsiflexors and plantarflexors and gross motor function was analyzed using the spearman’s correlation. pvalue< 0.05 was considered significant at 95% confidence interval. SPSS-13 version was used for analysis.
RESULTS
The aim of this study was to establish a relationship between spasticity, strength and gross motor function in children with spastic diplegia between the age group of 5 to 15 years. The subjects were classified on the gross motor function classification system levels I to V. The total number of children included in the study was 30, out of which 21 were males and 9 were females. The demographic data such as age and gender was obtained. The number of subjects mean and standard deviation of their age and GMFM scores is shown in table1 and figure 1. The comparison of spasticity and strength between the right and left sides showed that there was no significant difference in spasticity and strengthof hip flexors, extensors, adductors and abductors, knee flexors and extensors, ankle dorsiflexors and plantarflexors between the right and left side. Thus as observed that there was no significant difference in spasticity and strength of hip flexors, extensors, adductors and abductors, knee flexors and extensors, ankle dorsiflexors and plantarflexors between the right and left side.Therefore, the values of spasticity and strength of individual muscles of right and left side was added together for the purpose of correlation analysis between spasticity, strength and gross motor function. In addition, the aggregate values of spasticity and strength of each joint as a whole of right and left side together was correlated with gross motor function.
A. Correlation between spasticity, strength of muscles around the hip jointand gross motor function.
Correlation of spasticity and strength of muscles around the hip joint and gross motor function was done using the spearman’s correlation coefficient as shown in table 2 Correlation between Spasticity and GMFM As shown in table 2, there was a negative correlation of quality of muscle reaction of hip flexors and adductors, and positive correlation of angle of muscle reaction of hip flexors and adductors with GMFM. There was no correlation between the quality and angle of muscle reaction of hip extensors and abductors with GMFM. When aggregate spasticity of muscles around the hip joint was considered, there was negative correlation between quality of muscle reaction and positive correlation between angle of muscle reaction with GMFM as shown in table 2. Correlation between Spasticity and Strength The quality of muscle reaction of the hip flexor spasticity showed no correlation with strength but the angle of muscle reaction of hip flexors showed negative correlation with strength. There was no correlation between the quality and angle of muscle reaction of the hip extensors, adductors and abductors, and the aggregate hip joint with strengthas shown in table 2.
Correlation between Strength and GMFM
There was no correlation between the strength of hip flexors, extensors and adductors with GMFM. There was a significant positive correlation of hip abductor and the aggregate hip joint strength with GMFM as shown in table 2.
B. Correlation between spasticity, strength of muscles around the knee joint and gross motor function.
Correlation of spasticity and strength of muscles around the knee joint and gross motor function was done using the spearman’s correlation coefficient as shown in table 3. Correlation between Spasticity and GMFM As shown in table 3 there was no correlation between quality and angle of muscle reaction of knee flexors with GMFM.There was a negative correlation between the quality of muscle reaction of the knee extensors, and the aggregate of knee joint with GMFM and a positive correlation of the angle of muscle reaction of knee extensors and aggregate of knee joint with GMFM. Correlation between Spasticity and Strength There was no correlation between quality and angle of muscle reaction of the knee flexors, knee extensors and aggregate knee joint with its strength as shown in table 3 Correlation between Strength and GMFM There was a positive correlation between strength of knee flexors, and aggregate knee joint with GMFM. Strength of knee extensors had no correlation with GMFM as shown in table 3.
C. Correlation between spasticity, strength of muscles around the ankle joint and gross motor function
Correlation of spasticity and strength of muscles around ankle joint and gross motor function was done using the spearman’s correlation coefficient as shown in table 4. Correlation between Spasticity and GMFM As shown in table 4 there was no correlation in quality and angle of muscle reaction of ankle dorsiflexors with GMFM. There was a positive correlation between angle of muscle reaction of plantarflexors and aggregate ankle joint, and a negative correlation between the quality of muscle reaction of the aggregate ankle joint with GMFM. Correlation between Spasticity and Strength There was no correlation of quality and angle of muscle reaction of dorsiflexors, plantarflexors and aggregate ankle joint with its strength as shown in table 4. Correlation between Strength and GMFM There was positive correlation of strength in dorsiflexors, plantarflexors and aggregate ankle joint with GMFM as shown in table 4.
DISCUSSION
The aim of the current study was to establish a relationship between spasticity, strength and gross motor function of children with spastic diplegia of GMFCS-EandR levels I to V and to evaluate if individual joint involvement can have an effect on gross motor function. The spasticity was tested using the Tardieu scale, strength using the hand held dynamometer and gross motor function using the GMFM-88. The study included 30 children (21 males and 9 females) with a mean age of 9.17±3.26 years. Correlation between Spasticity and GMFM The results of the present study showed that the quality of muscle reaction of the hip flexors and adductors, knee extensors and aggregate of hip, knee and ankle was negatively correlated with the gross motor function. This shows, when spasticity is less, the level of gross motor function is high. In children with spastic diplegia, the spasticity in the lower extremities causes use of limited synergies in all function i.e. they have poor ability to grade co-contraction, reciprocal inhibition, terminate sustained muscle activity and abnormal timing of muscle contraction. In addition to these, there is rapid development of secondary impairments in the musculoskeletal system leading to shortened muscles, mal-aligned jointsand deformed bones. Shortening of hip flexors, adductors and medial hamstrings leads to hip flexion, adduction and internal rotation posture which narrows the base of support in all positions, and makes learning upright trunk control more difficult. When the flexion component is stronger than the internal rotation component, the rectus femoris exerts a strong influence as hip flexors and knee extensor leading to extended knees.5,9,27,33 Our results were supported by a report of Tuzson et al. who determined a spastic threshold velocity, using electromyography during isokinetic testing, for the quadriceps and hamstrings and found it moderately correlated with the Gross Motor Function Measure (GMFM) walk and run domain (r =.58) and walking velocity (r =.64), indicating the milder the spasticity, the higher the function.27 The ankle mostly assumes a plantarflexed position, because of the large cross-sectional area of the plantarflexors compared to the dorsiflexors. Functionally the child is limited to the use of hip flexion, adduction, internal rotation and ankle plantarflexion with variable knee positions. This leads to lack of variety of movement which is essential for gross motor function. Thus, when spasticity in certain muscle groups is high the gross motor function is reduced.4, 5 Correlation between Spasticity and Strength The results of the present study showed that the quality and angle of muscle reaction of hip extensors, adductors and abductors, knee flexors and extensors, ankle dorsiflexors and plantarflexors, had no correlation with strength except the angle of muscle reaction of hip flexor which was negatively correlated with strength.
The negative correlation between spasticity and strength of hip flexors could be because typically children with spastic diplegia use the superficial lumbar extensors with the hip flexors as prime movers or in synergy to control the upper trunk on the pelvis. As these children also have loss of selective motor control, so whenever there is a voluntary contraction of the muscles, the lumbar spine goes into end- range extension with some degree of hip flexion leading to overlengthened abdominal muscles further weakening their use. This repeated use of hip flexors which causes an increase in their strength and, therefore leads to a decrease in the spasticity. This was supported by Morton et al who reported that with an increase in the muscle strength there is decrease in the spasticity. This explains the negative correlation between spasticity and strength.5, 11, 28, 29 Correlation between strength and GMFM There was positive correlation between strength of hip abductors, knee flexors, ankle dorsiflexors and plantarflexors and aggregate hip, knee and ankle joint with GMFM. Hip abductors strength in children with CP has been reported to be significantly less than the typically developing children.Hip abductor strength is important for kneel and half kneel skills, single-limb balance, and gait. Weak hip abductors will result in a contralateral pelvic drop and excessive hip adduction during gait. In the present study, the hip abductor strength appeared to be most closely correlated to gross motor function than any other lower-extremity muscle group.5, 7, 10, 24 In the present study, there was a positive correlation between strength of knee flexors and GMFM. Most of the children with spastic diplegia have a crouched posture due to inability to terminate the hamstrings. Over a period, this constant over activity leads to shortening of hamstrings causing inefficient muscle strength and decrease in knee extension ranges. Most of the components of GMFM requires synergistic action of muscles across the joints with normal timing of contraction and graded control which becomes difficult with abnormal alignment because of the short hamstrings. This can explain the positive correlation of knee flexors and GMFM.5, 7, 18, 24 Strength of ankle dorsiflexors showed a highly significant correlation with gross motor function. This could be because children with spastic diplegia experience varying degree of reduced motor control of their distal musculature which compromises their ability to dorsiflex their feet. This presents a significant problem for many children as it impedes a normal heel strike during gait, forcing children to walk on toes. Various mechanisms contribute to reduced dorsiflexion including altered neural control, increased stiffness of the plantarflexors or weakness of dorsiflexors. 5, 15, 30 Similarly even plantarflexors showed positive correlation to gross motor function. Like the hamstrings, the plantarflexors are also overactive, leading to shortening and inability to move ankle joint effectively during functional activities. This weakness in ankle dorsiflexors and over activity in plantarflexors leads to inefficient co-contraction or timing causing reduction in standing and walking function. These results were similar to the study done by Ross et al who reported that increase in ankle strength improved gross motor function.6, 15, 31 In the present study a positive correlation was found between strength of hip abductors, ankle dorsiflexors and plantarflexors. Similar results have been shown in a previous study done by Ross et.al on relationship between spasticity, strength, gross motor function and gait in children with spastic diplegia. This study reported that strength was positively correlated with gross motor function in hip abductors, ankle dorsiflexors and plantarflexors.10 Subgroup analysis of relationship between spasticity, strength and gross motor function of children in the different levels of GMFCS was not done due to insufficient sample size in each level.
Psychometric properties of Tardieu scale in hip flexors, extensors, and abductors, knee extensors, have not been established. A single observer assessed all the children on the Tardieu scale, hand-held dynamometer and GMFM. This may have affected the results in the present study. Further studies can be done to find out relationship between spasticity, strength and gross motor function in other types of cerebral palsy and in different levels of GMFCS levels may be established.
CONCLUSION
In the relationship between spasticity and GMFM, there was a significant negative correlation between aggregate spasticity of muscles around the hip, knee and ankle joint and the gross motor function. But when individual muscle analysis was done only hip flexors, adductors and knee extensors showed negative correlation with gross motor function. In the relationship between strength and GMFM, a significant positive correlation was found between aggregate strength of muscles around the hip, knee and ankle joint and the gross motor function. But when individual muscle analysis was done only strength of hip abductors, knee flexors, ankle dorsiflexor and plantarflexors showed positive correlation with gross motor function. In the relationship between spasticity and strength, the spasticity of hip flexors were the only muscles that showed a negative correlation with strength, where as spasticity of other muscles showed no correlation with strength.
ACKNOWLEDGEMENT
Authors acknowledge the immense help received from the scholars whose articles are sited 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.
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7. Damiano D, Abel MF. Functional outcome of strength training in spastic cerebral palsy. Arch Phys Med Rehab1998;79:119-25.
8. Hanna SE, Palisano RJ, Rosenbaum PL, Russell DJ, Walter SD, Wood EP, Raina PS, Galuppi BE. Validation of a model of gross motor function for children with cerebral palsy. Phys Ther 2000;80:974-85
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10. Ross SA, Engsberg JR .Relationship between spasticity, strength, gait, and the GMFM-66 in person with spastic diplegic cerebral palsy. Arch Phys Med Rehab.2007;88:1114-20 .
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12. Eek MN .Muscle strength, gross motor function and gait pattern in children with cerebral palsy. Institution of clinical science \ Department of pediatrics at Schlgrenska academy, University of Gotherberg - 2009 – unpublished thesis
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareFORMATION OF MEDIAN NERVE BY MULTIPLE LATERAL ROOTS
English123128Sharadkumar Pralhad SawantEnglish Shaguphta T. ShaikhEnglish Rakhi Milind MoreEnglishAim: To study the formation of median nerve by multiple lateral roots. Materials and Methods: 50 upper limbs of 25 donated embalmed cadavers (20 males and 5 females) were studied in the department of Anatomy at K. J. Somaiya Medical College, Sion, Mumbai, INDIA, the formation of median nerve by multiple lateral roots were observed. The finding was noted after thorough and meticulous dissection of the upper limbs of both sides (axilla, arm, forearm and palm) of 25 cadavers. The photographs of the variations were taken for proper documentation. Observations: The variations were found in two cases. In the first case the median nerve was formed by multiple roots- three roots from lateral cord and one root from medial cord of brachial plexus. In the second case out of the three roots forming the anomalous median nerve, two were from lateral cord and one from medial cord of brachial plexus. However, the distribution of the anomalous median nerve was normal in arm, forearm and palm. The arterial pattern in the arm (axillary and brachial arteries) was also normal in both the specimens. Conclusion: The presence of such type of variations are clinically important for surgeons, orthopaedicians and anaesthetist performing pain management therapies on the upper limb.
EnglishAxilla, Brachial Plexus, Median Nerve, Multiple Roots, Pain Management Therapy.INTRODUCTION
The median nerve is normally formed by the union of lateral root of median nerve arising from the lateral cord (C5, C6, C7) of brachial plexus and medial root of median nerve arising from the medial cord (C8, T1) of brachial plexus anterior to the axillary artery. Some fibres from C7 often leave the lateral root to join the ulnar nerve. Clinically they are believed to be mainly motor to the flexor carpi ulnaris. The median nerve enters the arm at first lateral to the brachial artery. Near the insertion of the coracobrachialis, it crosses in front of the artery, descending medial to it, to the cubital fossa, where it is posterior to the bicipital aponeurosis and anterior to the brachialis. It usually enters the forearm between the heads of the pronator teres, crossing to the lateral side of the ulnar artery and separated from it by the deep head of pronator teres (1). Anomalous pattern of median nerve can be explained on the basis of embryological development The upper limb buds lie opposite the lower five cervical and upper two thoracic segments. As soon as the buds form, the ventral primary rami of the spinal nerves penetrate into the mesenchyme of limb bud. Immediately the nerves enter the limb bud, they establish intimate contact with the differentiating mesodermal condensations and the early contact between nerve and muscle cells is a prerequisite for their complete functional differentiation (2). The growth as well as the path finding of nerve fibres towards the target is dependent upon concentration gradient of a group of cell surface receptors in the environment (1). Several signalling molecules and transcription factors have been identified which induce the differentiation of the dorsal and ventral motor horn cells The high percentage of anomalies as mentioned above emphasizes the complexities and irregularities of this anatomic region with regard to surgical approaches (3). Knowledge of such variations is important for surgeons to perform surgical procedures in the axillary region and arm (4). Considering the high percentage of anomalies in the formation of median nerve and its paramount clinical importance, the present variations are documented.
MATERIALS AND METHODS
50 upper limbs of 25 donated embalmed cadavers (20 males and 5 females) were studied in the department of Anatomy at K. J. Somaiya Medical College, Sion, Mumbai, INDIA, the formation of median nerve by multiple lateral roots were observed. The findings were noted after thorough and meticulous dissection of the upper limbs of both sides (axilla, arm, forearm and palm) of 25 cadavers. Arterial pattern of upper limb was also noted. Photographs of the abnormalities were taken for proper documentation.
OBSERVATIONS
In the present study, the variations related to the formation of median nerve were found in two cases. In the first case the median nerve was formed by multiple roots- three roots from lateral cord and one root from medial cord of brachial plexus. In the second case out of the three roots forming the anomalous median nerve, two were from the lateral cord and one from the medial cord of brachial plexus. However, the distribution of the anomalous median nerve was normal in arm, forearm and palm. The arterial pattern in the arm (axillary and brachial arteries) was also normal in both the specimens.
DISCUSSION
Neural variations of the brachium constitute an important anatomical and clinical entity. Although frequently reported, if accompanied by other anomalies, they deserve special mention in anatomical literature. The nerves of the extremities are especially vulnerable to injury because of their long course and superficial distribution. The variations related to the formation of median nerve were found in literature. The variations related to the formation of median nerve by more than two roots are relatively uncommon as compared to the other types of variations of median nerve. Most of the variations were related to anomalous relationship between median and musculocutaneous nerves. As found in literature, it was documented that the lateral root was small and the musculocutaneous nerve was connected with median nerve in the arm (5). Another study involving dissection of ten cadavers,mentioned failure of separation of musculocutaneous nerve from the median nerve and the latter therefore gave off the branches that should arise from musculocutaneous nerve, namely branches to coracobrachialis, biceps brachii and major part of brachialis (6). The variations related to the formation of median nerve by more than two roots which have been observed in the present study are rare as revealed by survey of literature. These variations can be explained on the embryological basis. The first indication of limb musculature is observed in the seventh week of development as condensation of mesenchyme near the base of the limb buds. With further elongation of the limb buds, the muscle tissue splits into flexor and extensor compartments. The upper limb buds lie opposite the lower five cervical and upper two thoracic segments. As soon as the buds form, ventral primary rami from the spinal nerves penetrate into the mesenchyme. At first, each ventral ramus divides into dorsal and ventral branches, but soon these branches unite to form named peripheral nerves which supply extensor and flexor group of muscles respectively. Immediately after the above mentioned rearrangement of nerves, they enter the limb buds and establish an intimate contact with the differentiating mesodermal condensations and this early contact between the nerve and muscle cells is a prerequisite for their complete functional differentiation (2). Over the years, two principal theories have emerged concerning the directional growth of nerve fibres – the neurotropism or chemotropism hypothesis of Ramon et al (7) and the principle of contact-guidance of Weiss (8). The salient feature of chemotropism is that axonal growth cones act as sensors to concentration gradients of molecules in the environment and grow up the gradient towards the source, i.e. the target. There is no doubt, however that contact guidance mechanisms operate in parallel with neurotropism. Adhesion to the structures with which the growth cone contacts also plays a role. A group of cell surface receptors viz. neural cell adhesion molecule (N-CAM) and L1 and the Cadherins act as transcription factors which recognize and bind to components of the extracellular matrix. Thus, both cell-cell and cellmatrix interactions may be involved in axonal path finding (1). Over or under expression of one or multiple transcription factors as mentioned above have been found to be responsible for the variations in the formation, relation and distribution of the motor nerve fibers (1). Chauhan et al strongly recommend the consideration of the phylogeny and the development of the nerves of the upper limb for the interpretation of the nerve anomalies of the arm (9). The presence of such communications may be attributed to random factors influencing the mechanism of formation of limb muscles and the peripheral nerves during embryonic life. Significant variations in nerve patterns may be a result of altered signaling between mesenchymal cells and neuronal growth cones (10) or circulatory factors at the time of fusion of brachial plexus cords (11). Iwata believed that the human brachial plexus appears as a single radicular cone in the upper limb bud, which divides longitudinally into ventral and the dorsal segments. The ventral segments give roots to the median and the ulnar nerves with musculocutaneous nerve arising from the median nerve. He further kept the possibility of failure of the differentiation as a cause for some of the fibers taking an aberrant course as a communicating branch (12). Chiarapattanakom et al are of the opinion that the limb muscles develop from the mesenchyme of local origin, while axons of spinal nerves grow distally to reach the muscles and/or skin. They blamed the lack of coordination between the formation of the limb muscles and their innervation for appearance of a communicating branch (13). Anastomosis between the musculocutaneous nerve and the median nerve is by far the commonest and frequent of all the variations that are observed among the branches of the brachial plexus (14). Li Minor categorized these communications into following five type: In type I, there is no communication between the median nerve and the musculocutaneous nerve, in type II, the fibers of the lateral root of the median nerve pass through the musculocutaneous nerve and join the median nerve in the middle of the arm, whereas in type III, the lateral root fibers of the median nerve pass along the musculocutaneous nerve and after some distance, leave it to form the lateral root of the median nerve. In type IV, the musculocutaneous nerve fibers join the lateral root of the median nerve and after some distance the musculocutaneous nerve arises from the median nerve. In type V, the musculocutaneous nerve is absent and the entire fibers of the musculocutaneous nerve pass through the lateral root and fibers to the muscles supplied by musculocutaneous nerve branch out directly from the median nerve (15). The most frequent variation is the presence of a communicating branch that emerges from the musculocutaneous nerve and goes distally to join the median nerve, an anastomosis observed in the lower third of arm (16). If this branch is given off in upper third of the arm, it is generally considered as third (double lateral) root of the median nerve (16). In the present case, the musculocutaneous nerve in upper third of the arm, passed medially downwards and joined the median nerve. It can be considered as the double lateral root of the median nerve or in other words the median nerve can be said to be formed by three roots: a) one from the lateral cord; b) one from the musculocutaneous nerve; c) and the third from the medial cord. Similar variation was observed earlier by different authors - The median nerve, instead of having two roots may have three roots - either one each from lateral cord, medial cord and musculocutaneous nerve (9) or two from lateral cord and one from the medial cord (17) or it may have even four roots – three from the lateral cord and one from the medial cord (18). Knowledge of anatomical variations of these nerves at the level of upper arm is essential in light of the frequency with which surgery is performed in the axilla and the surgical neck of the humerus (19). The variations as noted in the present study may be attributed to over or under expression of one or more transcription factors as mentioned above. The variations in the formation and relations of median nerve in the arm bear remarkable clinical significance. Considering these variations Rao advocated that the clinicians and surgeons should be aware of such variations while performing surgical procedure in this region (20). Injury to such a variant nerve in the proximal arm may lead to a galaxy of manifestations including sensory, motor, vasomotor and trophic changes (21). The possible clinical implications of these variations relating either to the surgical approach to the shoulder joint and entrapment syndromes are important (22). Anomalies of axillary or brachial artery are frequently related to unusual pattern of brachial plexus and median nerve (23). However, in the present study no abnormal arterial pattern was detected.
Clinical Significance
The anatomical variation described here has practical implications, since injury to the median nerve in the axilla or arm would, in this case, have caused unexpected paresis or paralysis of the flexor musculature of the elbow and hypoesthesia of the lateral surface of the forearm, in addition to the classical signs that are already well known. Injury to the median nerve could occur in cases of open or closed trauma to the arm, such as bullet and blade wounds or during surgeries on the axilla or arm. The median nerve and its roots are close to the axillary vein, which is used as the most cranial limit for axillary lymph node dissection, a procedure used in treating certain tumors, such as breast carcinoma and melanoma. If the dissection extends more cranially than normal, injury to the median nerve (or to its medial root) may occur, with consequent dysfunction of the flexor musculature of the elbow if the anatomical variation described here is present. It would not be unlikely for such accidents to occur even with the most eminent surgeons, considering that the classical concept is that the median nerve does not give rise to branches in the arm (24).
CONCLUSION
The variations in the formation of median nerve by multiple lateral roots are clinically important for surgeons, orthopaedicians and anaesthetist performing pain management therapies on the upper limb. These variations are compered with the earlier data and it is concluded that variations in branching pattern of cords of brachial plexus are a rule rather than exception. A lack of awareness of variations with different patterns might complicate surgical repair and may cause ineffective nerve blockade.
Competing Interests:
The authors declare that they have no competing interests.
Authors' contributions:
SPS wrote the case report, performed the literature review and obtained the photograph for the study. RMM performed the literature search and assisted with writing the paper. STS conceived the study and helped to draft the manuscript. All authors have read and approved the final version manuscript.
ACKNOWLEDGEMENT
All the authors wish to convey our sincere thanks to Dr. Arif A. Faruqui for his valuable help, support and inspiration. Authors also 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=2005http://ijcrr.com/article_html.php?did=20051. Williams P.L., Bannister L.H., Berry M.M. et al. Gray’s Anatomy. In: Nervous System. 38th ed. London Churchill Livingstone, 1999: 231- 232,1270.
2. Saddler T.W. Langman’s Medical Embryology. In: Muscular system. 10th ed. Philadelphia Lippincott Williams and Wilkins, 2006: 146-147.
3. Edglseder WA JR, Goldman M. Anatomic variations of the musculocutaneous nerve in the arm. Amer J Orthop 1997; 26: 777-80.
4. Uysal II, Seker M, Karabulut AK, Buyukmumcu M, Ziylan T. Brachial plexus variation in human foetuses. Neurosurgery 2003; 53: 676-84; discussion 684.
5. Standring S, Ellis H, Healy JC, Johnson D et al. Gray’s Anatomy. In: General organisation and surface anatomy of the upper limb.39th ed. Philadelphia Elsevier Churchill Livingstone, 2005: 803-4.
6. Guha R, Palit S. A rare variation of anomalous median nerve with absent musculocutaneous nerve and high up division of brachial artery. J Interacad 2005; 9: 398-403.
7. Ramon Y, Cajal S. The neurotropism or chemotropism hypothesis. Trab Lab Invest Biol 1919; 17: 65-8.
8. Weiss P. Nerve patterns: the mechanics of nerve growth. Growth (suppl 5) 1941; 163- 203.
9. Chauhan, R., Roy, TS. Communication between the median and musculocutaneous nerve: A case report. Journal of Anatomical Society of India, 2002, vol. 51, n. 1, p. 72-75.
10. Abhaya, A., Bhardwaj, R., Prakash, R. Dual origin of musculocutaneous nerve. Journal of Anatomical Society of India, 2003, vol. 52, n. 1, p. 94.
11. Kosugi, K. Mortia, T., Yamashita, H. Branching pattern of the musculocutaneous nerve. 1. Cases possessing normal biceps brachii. Jikeakai Medical Journal, 1986, vol. 33, p. 63-71.
12. Iwata, H. Studies on the development of the brachial plexus in Japanese embryo. Republic Department Anatomy Mie Prefect University School of Medicine, 1960, vol. 13, p. 129-144.
13. Chiarapattanakom, P., Leechavengvons, S., Witoonchart, K., Uerpairojkit, C., Thuvasethakul, P. Anatomy and internal topography of the musculocutaneous nerve: The nerves to the biceps and brachials muscle. Journal of Hand Surgery, 1998, vol. 23A, p. 250-255.
14. Venieratos, D., Anagnostopoulou, S. Classification of communications between the musculocutaneous and median nerves. Clinical Anatomy, 1998, vol. 11, p. 327-331.
15. Li Minor, JM. A rare variant of median and musculocutaneous nerves in man. Archives Anatomy Histology Embryology, 1992, vol. 73, p. 33-42.
16. Bergman, RA., Afifi, AK., Miyauchir, RA. Ilustrated encyclopedia of human anatomic variation. In: NERVOUS system - plexuses. 1988. Available from: http://virtualhospital.com. Universityofiowacare.
17. Mohapatra, BB., Chinara, PK., Dutta, BK., Nayak, AK. Variation in the formation and branching pattern of median nerve. Journal of Anatomical Society of India, 2004, vol. 53, n. 1, p. 31-66.
18. Uzun, A., Seelig, LL. A variation in the formation of the median nerve: communicating branch between the musculocutaneous and median nerves in man. Folia Morphologica (Warsz), 2001, vol. 60 , n. 2, p. 99-101.
19. Leffert, RD. Anatomy of the brachial plexus. New York: Churchill Livingstone, 1985. 384 p.
20. Rao PPV, Chaudhary SC. Communication of Musculocutaneous nerve with the median nerve. East Afr Med J 2000; 77: 498-503.
21. Saeed M, Rufai AA. Median and musculocutaneous nerves: variant formation and distribution. Clin Anat 2003; 16: 453-7.
22. Venieratos D, Anagnostopoulou S. Classification of communication between musculocutaneous and median nerves. Clin Anat 1998; 11: 327-31.
23. Basar R, Aldur MM, Celik HH, Yuksel M, Tascioglu AB. A connecting branch between the musculocutaneouos nerve and the median nerve. Morphologie 2000; 84: 25-7.
24. FregnanI, JHTG., Macéa, MIM., Pereira, CSB., Barros, MD., Macéa, JR. Absence of the musculocutaneous nerve: a rare anatomical variation with possible clinical-surgical implications. Sao Paulo Medical Journal, 2008, vol. 126, n. 5, p. 288-90. PMid:19099164.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesCORRELATION BETWEEN THE GLOBAL RADIATION AND SOLAR INDICES
English129132Sumathi.REnglish Samuel Selvaraj.REnglishThe relationship between Global radiation on a horizontal surface at Chennai and solar indices(smoothed sunspot number,10.7cm solar flux, aa index and sunspot area) are individually analyzed in this paper. Analysis has been done for the period from 2008-2010. The method used for the analysis has been Least square regression method. The relationship revealed that the Global radiation correlates negatively with the solar indices.
EnglishINTRODUCTION
Global solar radiation is an important parameter necessary for most ecological models and serves as an input for different photovoltaic conversion system, which is of greater Economic importance for renewable energy alternation. The solar radiation reaching the Earth?s surface depends on the climatic conditions of the specific site location. The sun is a constant energy source and is known for its cyclic behavior. Solar radiation output changes periodically and it affects the Earth and its atmosphere different ways , such as the formation of aurora, adverse effects on the satellite communications. Various indicators for the sunclimate connection such as sunspot numbers(Chambers,1878)[1],Solar cycle length(Friis-Chrislensen and Lassen.1991)[2] , Geomagnetic aa indexes(Cliver and Borier Koff,1998)[3] and Solar irradiance changes (Floyd et.al 2001)[4] have been reviewed in past literature. Recently Samuel selvaraj et.al 2011[5] have done research on the variation of Chennai global radiation due to sunspot number. W.E.Alnaser et.al(1991) developed a correlation study between Global radiation and sunspot number.[6] Earth receives a total amount of radiation by its cross section (π·RE²), but as it rotates this energy is distributed across the entire surface area (4·π·RE²). Hence the average incoming solar radiation (called the solar irradiance), taking into account the angle at which the rays strike and that at any one moment half the planet does not receive any solar radiation, is onefourth the solar constant (approx 342 W/m²). At any given moment, the amount of Solar radiation received at a location on the Earth's surface depends[10] on the conditions of the atmosphere and the latitude of the location. This Solar Irradiance hits the surface of the earth in two forms, beam (Gb) and diffuse (Gd). The beam component comes directly as irradiance from the sun[11,12], while the diffuse component reaches the earth indirectly and is scattered or reflected from the atmosphere or cloud cover.The total irradiance on a surface is G = GB + Gd Sunspots are temporary on the photosphere of the sun that appear visibly as dark spots compared to surrounding regions. The F 10.7 index is a measure of the solar radio flux per unit frequency at wavelength of 10,7cm, near the peak of the observed solar radio emission. The aa index is a measure of the disturbance level of the Earth?s magnetic field based on magnetometer observations at two nearly antipodal stations. The daily sunspot area is the measure of the area of solar surface covered by spots (in the sunspot area unit namely ppm of solar hemisphere) in an observational day. The monthly average of the daily sunspot area usually bears information of long term variations of solar activity.[7,8] As the number of spots increases the total area of the spots naturally increases and their magnetic complexity usually grows as well. The objective of the work is to look for individual correlation between the monthly averaged daily global radiation on a horizontal surface at Chennai with solar indices like sunspot number .10.7cm solar flux , aa index and sunspot area . Method of least squares is used to find out the Mean Bias Error , Root Mean Square Error and Mean Average Percentage Error.
DATA ANALYSIS AND METHODOLOGY
The local climate was analysed using the following data. The monthly average daily global radiation on a horizontal surface for Chennai for 30 months(July2008 to December2010) were obtained from the Indian Metrological Department, Pune. The solar indices data were taken from the site ftp://Ftp.ngdc.noaa.gov/STP/SOLAR_DATA.
The concept of „correlation? is a statistical tool which studies the relationship between two variables. “Regression analysis is a mathematical measure of the average relationship between two or more variables in terms of the original units of the data.”Regression analysis, in general sense, means the estimation or prediction of the unknown value of one variable from the known value of the other variable. For this study, correlation coefficients are calculated to analyze the statistical relationship between global radiation and solar indices like sunspot number,10.7cm solar flux, aa index and
sunspot area. Correlation coefficient for global radiation with solar indices have been found individually for every index. Table 2 shows the comparison of the correlation coefficient of Global radiation with the sunspot number,10.7cm solar flux, aa index and sunspot area Sunspot number,10,7cm Solar flux,sunspot area are negative correlation with the Global radiation,but aa index show no significant correlation with the Global radiation. After performing all the Regression techniques to study the correlation between global radiation and the solar indices, it was found that the linear model works with least error. The linear regression equation for solar indices are Y=aX + b Where Y-Global radiation, X-Solar indices, and a,b-regression coefficients obtained by fitting measured data. The following statistical errors are used to test the performance of model.[9] Mean Bias Error is defined as MBE=(1/N)∑(Hcomputed –Hmeasured) Where N denotes total number of observations. This test provides an information on the long term performance. A low MBE is desired. Ideally a zero value of MBE should be obtained. A positive value gives the average amount of over-estimation in the calculated value or vice versa. One draw back of this test is that over estimation of an individual?s observation will cancel under estimation in a separate observation. Root Mean Square Error is defined as RMSE={[∑(Hcomputed - Hmeasured)2]/N}1/2 Root mean Square Error is always positive, a zero value is ideal. This test provides information on the short term performance of the models by allowing a term by term comparison of the actual deviation between the calculated value and the measured value. Mean percentage error is defined as MPE=[∑(Hmeasured – Hcomputed/Hmeasured) X 100]/N
RESULT AND CONCLUSION
The monthly average daily global radiation by the linear model have been compared with solar indices . The present study shows that linear model between global radiation and sunspot number can result in low error level. MPE is 9.54298, RMSE is 1.661619 and MBE is 0.67995,followed by the 10.7solar flux MPE 10.22973 ,RMSE 1.815245, MBE 0.69509 ,then aa index MPE 13.99744, RMSE 2.828876, MBE 2.104048 and the sunspot area MPE 12.04038 RMSE 9.591663 MBE -0.01392. So sunspot number is more correlated with Global radiation as there is least error in comparison to other indices.
Englishhttp://ijcrr.com/abstract.php?article_id=2006http://ijcrr.com/article_html.php?did=20061. Chambers,F.,(1878),?Sunspots and Weather?, Nature 18, p.567-568.
2. Friis-christensen,K.Lassen(1991)?Length of solar cycle: An indicator of solar activity closely associated with climate? Science Vol.254,p.698-700.
3. Cliver EM.Boriakoff V.,Feynman,J.,(1998),;Solar variability and climate change: Geomagnetic aa index and global surface temperature? Geophysical Research Letter Vol.25,P.1035-1038.
4. Floyd.L.D.,Prinz,P.C.Crane,and L.C.Herring(2001),?Total solar and spectral irradiance Variations from solar cycles 21 to 23? Advanced Space Research in press.
5. Samuel Selvaraj.R and Sivamadhavi.V(2011),?Solar insolation at Chennai during the 23rd Solar cycle? Universal Journal of Environmental Research and Technology,Vol3,381-384.
6. Alnaser,W.E.,and M.J.Alothman(1991)?Studies of the correlation between the Global radiation, Ultraviolet Radiation ,Cosmic Radiation,Sunspotnumber and Meteorological Parameters?Astrophysics and space science,Vol.186,P.191-203.
7. A.P.Sarychev and E.M Rosh china,?Total sunspot area as a Solar activity index? Solar System Research,Vol.40,No.6 521-526 (2006).
8. Yu.A. Nagovitsyn „To the description of longterm variations in the solar magnetic flux; The sunspot area index? Springer link,Astronomy letter Vol.31,No.8 557-562(2005).
9. M.Sekae,M.Sakthivel,s.Satheesh Kumar,C,Ramesh(2012),?Estimation of Global Solar Radiation for chennai?,European Journal of Scientific Research?,Vol.73,P.415-424.
10. Smart,W.M. (1980),Text book on Spherical Astronomy(6th edition),Cambridge University press,Cambridge,P-69.
11. Alam,M.S.,Saha,S.K.,Chowdhury,M.A.K.,Saif uzzaman and Rahman,M.,(2005), ?Simulation of solar radiation system? American.Journal ofApplied science 2(4),P.751-58
12. Badescu,V.,(1999),?Correlations to estimate monthly mean daily solar global irradiation Application to Romania?,Energy 24 ,P.883- 893.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareSEX DETERMINATION BY MASTOID PROCESS IN SOUTH INDIAN POPULATION
English133137Suresh SukumarEnglish Sushil YadavEnglish Vimal Kumar MEnglish Manju H BEnglishObjective: The purpose of this study was to evaluate the significance for sex determination of the measurement of the area formed by projection of 3 craniometric points related to the mastoid process: the porion, asterion, and mastoidale points. Method: Fifty skulls, 25 males and 25 females, were analysed. The three craniometric points were located and marked on both side of the skull and the measurement was done by digital calliper (0.01mm).The area of mastoid triangle was calculated by means of the Heron’s formula. Total area calculated was done by adding the areas (mm2) obtained on each side. Result:From this study the areas of the mastoid triangles in male skulls 1611.747 mm2 > areas of the mastoid triangles in female skulls1 241.13 mm2. Minimum area of mastoid in male Right Area (mm2) is 702.36 and left area is 680.16 with Total Area (mm2) 1411.08. Maximum area of mastoid, in male Right Area (mm2) is 991.84and left area is 1008.25with Total Area (mm2) 2000.09. . Minimum area of mastoids, in female Right Area (mm2) is 506.56and left area is 468.81with Total Area (mm2) 982.58. Maxium aread of mastoiod in female Right Area (mm2) is 857.35left area is 848.16with Total Area (mm2) 1705.51
EnglishINTRODUCTION
Historically, human identification is one of the most challenging subjects that man has confronted. In the skull, the temporal bone is highly resistant to physical damage1 ; thus it is commonly found as remainder in skeletons that are very old; this study examines patterns of variation within a group, particularly the variation between sexes. The topic of this study is the mastoid process, which can be defined as a coneshaped process of the temporal bone located posterior to the external auditory meatus that projects inferiorly. Pelvis is one of the best area to determine the sex, but most of the time it is damage. The most obvious example being the distinctive appearances of the android and gyneacoid pelvis5 . However, other skeletal appearances can be analysed including ossification centres, shapes of skull and mandible and the rarely used sternal length6 . Gender can be assessed by both anthropological and radiological means. In cases of skeletal remains, radiography may not be necessary if the features can be observed.Paiva andSegre (2003) introduce the technique is based on the triangular area calculation obtained between the points porion, mastoidale, and asterion, measured from xerographic copy of skulls2. From their result they had conculude that significant difference in area between the right and left mastoid triangle when comparing male and female skulls. According to Paiva andSegre (2003), When the total area was lower than or equal to 1260.36 mm2 , the skull is recognized as female skull2 and When it is higher than or equal to 1447.40 mm2 , the skull is recognized as male skull2 .The presence of sexual dimorphism in the mastoid triangle has been recently questioned4 .
OBJECTIVE
The purpose of this study was to evaluate the significance for sex determination of the measurement of the area formed by projection of 2 craniometric points related to the mastoid process: the porion, asterion, and mastoidale points.
METHOD
Fifty skulls, 25 males and 25 females, were analysed. The three craniometric points were located and marked on both side of the skull and the measurement was done by digital calliper (0.01mm).The area of mastoid triangle was calculated by means of the Heron’s formula. Total area calculated was done by adding the areas (mm2 ) obtained on each side. Statistical Analysis: The ‘Independent Samples t Test’ was used to evaluate the mean differences of the measured parameters. A comparison was performed between male and female and p areas of the mastoid triangles in female skulls1 241.13 mm2. Minimum area of mastoid, in male Right Area (mm2) is 702.36 and left area is 680.16 with Total Area (mm2) 1411.08. Maximum area of mastoid in male Right Area (mm2) is 991.84and left area is 1008.25with Total Area (mm2 ) 2000.09.Minium area of mastoid in female Right Area (mm2) is 506.56and left area is 468.81with Total Area (mm2) 982.58. Maxium aread of mastoiod in female Right Area (mm2) is 857.35left area is 848.16with Total Area (mm2) 1705.51. According to Paiva andSegre (2003), When the total area was higher than or equal to 1447.40 mm2 , the skull is recognized as male skull. According to Paiva andSegre (2003), when the total area was lower than or equal to 1260.36 mm2 , the skull is recognized as female skull. Table – 1 the areas of the female mastoid triangles about (about here) Table – 2 the areas of the male mastoid triangles (about here) Table 3, showing the areas of the mastoid triangles in male skulls (about here) Table 4, showing the areas of the mastoid triangles in female skulls (about here)
DISCUSSION
The analysis of the mastoid process characteristics is important in the determination of sex for forensic purposes. The mastoid region used in this study, being a part of the temporal bone, is recognized as being the most protected and resistant to damage, due to its anatomical position at the base of the skull. This has been demonstrated by Kloiber (1953), Wells (1960), Schäefer (1961), Gejval (1963), and Spence(1967), as cited by Wahl and Henke10(1980) According to Paiva andSegre (2003),When it is higher than or equal to 1447.40 mm2 , the skull is recognized as male skull. In present study when the skull were analyzed as described by Paiva andSegre, the values overlapping were about 20% for male skulls.According to Paiva andSegre (2003),When it is lower than or equal to 1260.36 mm2 , the skull is recognized as female skull. In present study when the skull were analyzed as described by Paiva andSegre, the values overlapping were about 32% for female skulls.
CONCLUSION
The area of the mastoid triangle can be used to determine the sex of the skull but the determination of sex can be better confined if the studies are carried out with greater number of skulls. Conflict of interest statement: We (the authors) confirm that there are no conflicts of interest associated with the submission of this article
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=2007http://ijcrr.com/article_html.php?did=20071. Kalmey, J. K. and Rathbun, T. A. Sex determination by discriminant function analysis of the petrous portion of the temporal bone. J. Forensic Sci., 41:865-7, 1996
2. De Paiva, L. A. and Segre, M. sexing the human skull through the mastoid process. Rev. Hosp. Clin. Fac. Med. São Paulo, 58:15-20, 2003.
3. Standring S, ed. Gray’s Anatomy. 40th Ed., Philadelphia, Elsevier, Churchill Livingstone. 2005.
4. Kemkes, A. and Gobel, T. Metric assessment of the "mastoid triangle" for sex determination: a validation study. J.Forensic Sci., 51:985-9, 2006
5. Walsh M, Reeves P, Scott S. When disaster strikes; the role of the forensic radiographer. Radiography 2004; 10:33-43.
6. Al Ekrish AA, Ekram M. A comparative study of the accuracy and reliability of multidetector computed tomography and cone beam computed tomography in the assessment of dental implant site dimensions. Dental Maxillofacial Radiol 2011; 40:67-75.
7. SUAZO, G. I. C.; ZAVANDO, M. D. A. and SMITH, R. L. Sex determination using mastoid process measurements in Brazilian skulls. Int. J. Morphol., 26(4):941-944, 2008
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN-0001November30HealthcareFORMULATION AND EVALAUTION OF SUSTAINED RELEASED AZITHROMYCIN MICROSPHERE BY EMULSION SOLVENT EVAPORATION TECHNIQUE
English138147Preeti. R. MeshramEnglish Suvarna P. GajareEnglish Manoj A. BhadaneEnglishAzithromycin is macrolide antibiotic is effective against a wide variety of bacteria such as Hemophilus influenzae, Streptococcus pneumoniae, Mycoplasma pneumoniae, Staphylococcus aureus, mycobacterium avium, and many others. The aim of present work was to formulate sustained released azithromycin microsphere to overcome the problem related to drug like insoluble in water, bitter taste and gastrointestinal side-effects. Microspheres were prepared by varying drug polymer ratio using emulsion solvent evaporation technique. Characterization of microspheres was done by Differential Scanning Calorimetry (DSC), X-Ray diffractometry (XRD), Surface Morphology (SEM), Stability studies.
EnglishSustained Released Microsphere, Azithromycin, Solvent evaporation technique, Ethyl Cellulose, Ethyl AcetateINTRODUCTION
The objective of any drug delivery system is to provide drug in therapeutic amount to the proper site in the body to achieve immediately and then maintain the desired drug concentration. These idealized objectives are achieved by appropriately developed sustained release drug delivery which also has diverse applicability and merits. There are certain drugs having large dose they may cause GI side effect so to avoid this drug can be formulated in sustained form to release slowly in lower GI tract. The sustained release drug delivery includes the application of physical and polymer chemistry. These polymers slowly release the drug in biosystem and maintain drug blood level within therapeutic range for longer duration. Some of the products characterize the drug permeation through the appropriate biological membrane and any first pass metabolic effects prior to the entry of drug into systemic circulation. The fact that the absorption and release rate of the drug from the dosage form, is one of the interesting and most recent developments in pharmaceutical field1, 2 . Microencapsulation is a process whereby relatively thin coating of polymers are applied to small particles of solid or droplets of liquid and dispersions. The microencapsulation processes produce small particles ranging in size from 1 to 1000 µm. There are different names for these particles: microparticle, microsphere, microcapsule and micromatrix. The microparticle system has become an indispensable part of the sustained drug delivery fields for the past few decades since it can readily be adapted for various administration methods3, 4, 5 .
MATERIALS AND METHODS
Procurement of drug and excipients: The drug Azithromycin and polymer Ethyl cellulose (50 cps) were gifted from Zim Laboratories Ltd, Nagpur, where other AR grade excipients like Polyvinyl alcohol, Dichloromethane, Chloroform, Ethyl acetate, Sodium hydroxide and Dibasic sodium phosphate were supplied by Loba chemical, Mumbai. Microspheres were prepared by using mechanical stirrer of Remi Motors Ltd, Mumbai. Evaluation and characterization of microspheres was carried out by using equipments like USP Tablet dissolution apparatus Type II (TDT 08L), UVVisible double beam Spectrophotometer (UV1800), FTIR (8400SCCE), X Ray diffractometer (PW1700), DSC (61000), Scanning Electron Microscope ( JSM 6380 A), Light Microscope and Stage Micrometer. METHODS Preliminary study 6, 7, 8: The Preformulation study like Preliminary identification test (melting point, solubility) characterization of drug and polymer and their interaction study were performed. Spectroscopic studies 9, 10, 11: UV Spectroscopy studies involved Determination of λ max and preparation of standard curve of Azithromycin at 244 nm. The IR spectrum of the drug sample was recorded by IR spectroscopy. The drug polymer interaction was carried out using FT-IR Apparatus. Formulation of microspheres: The formulations were prepared by using solvents ethyl acetate at different drug: polymer ratio (1:1, 1:1.5, and 1:2). Method of preparation 12, 13, 14: In the O/W emulsion solvent evaporation method, the polymer (ethyl cellulose) was dissolved in internal organic phase of ethyl acetate. Accurately weighed quantity 1g of Azithromycin was dispersed or dissolved in the polymer solution. This resulting mixture was poured slowly with stirring into 100 ml of a 0.015% w/v aqueous solution of polyvinyl alcohol. The emulsion was then stirred continuously at 700 rpm for 1 hr to evaporate the solvent. The microspheres were recovered by vacuum filtration, washed with 200 ml of deionized water and dried at room temperature. The compositions of Azithromycin microspheres using ethyl acetate as solvent were given in Table 1. Evaluation of microspheres15, 16: Angle of repose: Angle of repose is defined as the maximum angle possible between the surface of pile of powder and horizontal plane. The angle of repose for the microspheres of each formulation was determined by the funnel method. The microspheres were allowed to flow out of the funnel orifice on a plane paper kept on the horizontal surface. It forms a pile of microspheres on the paper. The angle of repose was calculated by substituting the values of the base radius ‘R’ and pile height ‘H’ in the following equation Bulk density of all batches of microspheres was determined by pouring gently 2 g of sample through a glass funnel into a 10 ml graduated cylinder. The volume occupied by the sample was recorded. Bulk density was calculated as per given formula:
Tapped Density:
The tapped density was determined by pouring 2 g of microspheres through a glass funnel into a 10 ml graduated cylinder. The cylinder was tapped from height of 2 inches until a constant volume was obtained. Volume occupied by the sample after tapping was recorded .The values for tapped density was calculated as per given formula:
Compressibility index:
The compressibility indices of the formulation blends were determined using Carr’s compressibility index formula.
Hausner Ratio:
It provides an indication of the degree of densification which could result from vibration of feed hopper. Lower the Hausner ratio better is the flowability. It was calculated as per given formula.
Determination of particle size 17:
The particle size was determined using stage micrometer. The diameters of about 300 microspheres were measured and the average particle size was determined. Estimation of drug loading18, 19: For determination of drug content, microspheres equivalent to 100 mg were weighed and dissolved in 100ml of acetone. After suitable dilutions were with phosphate buffer (pH 6.0), the resulting solution was analyzed spectrophotometrically at 244 nm.
In-vitro dissolution study20, 21, 22: The study was carried out using dissolution apparatus USP Type-I (Rotating Basket type). Accurately weighed microspheres equivalent to 200 mg of Azithromycin were taken in muslin cloth and it was kept in baskets. Dissolution study was carried out in phosphate buffer pH 6.0 at 50 RPM at temp 37 ± 0.5° c. During dissolution study 10 ml of aliquot was withdrawn at a time intervals of 1 to 12 hr and same was replaced with equal volume of fresh medium. The withdrawn samples were filtered through Whatmann filter paper and absorbances were measured at 244 nm. Drug concentration in the samples was determined from the calibration curve. Characterization of microspheres: Differential Scanning Calorimetry 9, 10, 11, 27:
The DSC measurements were performed on a differential scanning colorimeter with thermal analyzer. All accurately weighed samples (about 10 mg of Azithromycin, ethyl cellulose, physical mixtures, and formulation) were placed in a sealed aluminium pan, before heating under nitrogen flow (10 ml/min) at a scanning rate of 20 ° c per min from 100 to 300 ° c an empty aluminium pan was used as reference. X-Ray diffractometry 13, 14, 15, 23: The powder X-ray diffraction pattern of Azithromycin and polymer were obtained using Phillips X-ray diffractometer with a Ni-filtered CuKα-radiation at a scanning speed of 10 °/min at 2θ.The graph was plotted in 2 theta angle Vs intensity count. Surface Morphology 24 The microspheres were coated with Platinum by ion sputtering using Autofine coater. The microspheres were kept on the sample holder and the scanning electron micrographs were taken. Stability studies 35 The study was carried out by storing the microspheres in glass bottle 40 ° c and 75% RH for 30 days. These samples were collected on 7th, 14th , 21st, 28th day and analyzed for changes drug content and in-vitro dissolution studies. RESULTS Preformulation studies: The melting point of azithromycin was found in the range of 119 – 121 ° c. The wavelength of maximum absorbance (λ max) found was 244 nm. Evaluation of microspheres: All the formulations show angle of repose value in the range of 19.72 to 20.42. The values for bulk density were found in the range of 0.352 to 0.450. The values for tapped density were found to range from 0.414 to 0.510. The compressibility index values were found in the range of 11.76 to 14.99 respectively. Hausner’s ratio was ranging from 1.13 to 1.17. The average particle size of microcapsules is found to be within 151.263 to 171.342 µm. The % encapsulation efficiency is found to be in the range of 73.50 to 75. 60%. In – vitro dissolution study, % cumulative drug release from formulation in 12 hrs was decreased from 80.13 to 69.98 % represented in Table 2. The dissolution data for formulations C1, C2, and C3 was fitted to various drug release kinetic models like Zero order, First order, Higuchi Matrix and Korsemeyer Peppas, Hixon-Crowel model. Kinetic Treatment to Dissolution Data proved important information about rate constants (K), correlation coefficients (R) obtained for various models were tabulated in Table 3. Characterization of microspheres: The FT-IR spectra of pure drug - Azithromycin, ethyl cellulose, physical mixture and formulation (C1) shows major functional groups of Azithromycin (methyl substituted nitrogen on lactone ring) in spectra of individual drugs as well as in spectra of physical mixture and formulation (C1). FT-IR of Formulation C1 presented in Fig 1. DSC curve presented in fig 2 of Azithromycin shows a single endothermic peak at 121 ° c, due to melting of the drug. Optimized formulation C1 which contains Azithromycin and ethyl cellulose, the thermogram indicates characteristic peaks for melting of Azithromycin at 121 ° c. The XRD pattern presented in fig 3 of formulation exhibited halo pattern with less intense and denser peaks compared to plain Azithromycin, Scanning Electron Microscopy results were presented in fig 4a and 4b. Accelerated stability studies (AST) was carried for optimized batch (C1) by exposing it to 40° c/75% RH for one month and analyzed the samples at the interval of 7,14,21,28 days and the samples was analyzed for drug content and invitro dissolution study. The stability studies show drug content after 28 day was 74.14 where percent cumulative drug release 80.55. DISCUSSION The XRD scan of Azithromycin showed intense peaks of crystallinity. Diffractogram of azithromycin showed high intensity peaks between 2θ of 10-200 values demonstrating the crystalline nature of drug. No intense peaks were observed in diffractogram of ethyl cellulose which indicates amorphous nature. Thus, preformulation studies indicated that there is no interaction between Azithromycin and ethyl cellulose. Physical evaluation of microsphere formulation for angle of repose, Compressibility index and Hausner’s ratio showed that they had good flow properties. The microsphere formulated using ethyl acetate as internal organic phase or solvent in ratio1: 1 (C1) showed better encapsulation efficiency than other formulations. All the formulations showed diffusion exponent (n) varying from 0.524 to 0.668. From the n values of all formulations it can be concluded that as the concentration and viscosity of polymer was increased the value of diffusion exponent also increases. From the dissolution profile of formulations C1, C2 and C3, It can be concluded that formulation C1 shows better drug release (80%in 12 hrs) than C2 and C3. Formulations C1 to C3 showed retardation of Azithromycin as concentration of ethyl cellulose increased. Total release in 12 hrs was decreased from 80% to 70%. The IR data of Azithromycin loaded ethyl cellulose microsphere also showed the peaks same as that the peaks in IR spectrum of pure drug, this suggested that there was no chemical interaction between ethyl cellulose and Azithromycin. The DSC results also support IR spectrometry results indicating absence of drug - polymer interactions. The XRD pattern of formulation C1 exhibited halo pattern with less intense and denser peaks compared to plain Azithromycin. This indicated that Azithromycin is dispersed at the molecular level in the blend of polymeric matrix. The SEM photographs showed that the microspheres were spherical in nature and had a smooth surface. SEM photographs revealed the absence of crystals of drug on the surface of microspheres and uniform distribution of the drug within the microspheres.
The stability study conducted for observing effect of temperature on the formulation showed that there is no significant change in drug content and in-vitro drug release of optimized formulation. CONCUSION From the investigation carried out and results obtained, it was observed that the % of drug released from microspheres was sustained. As the GI side effects of Azithromycin are related to its high dose which can be minimized by formulating as microspheres. It would also helps to improve bioavailability because of absorption window and microencapsulation helps to mask the bitter taste of drug. Hence from this investigation, we can propose that the objective of the study is achieved. ACKNOWLEDGEMENT Authors are grateful to Zim Laboratoris Nagpur and Loba chemical, Mumbai, for providing the gift sample of drug and chemicals.
Englishhttp://ijcrr.com/abstract.php?article_id=2008http://ijcrr.com/article_html.php?did=20081. Brahmankar DM, Jaiswal SB. Biopharmaceutics and pharmacokinetics a treatise. 1st ed. New Delhi: Vallabh Prakashan Publications; 1995. p. 50-57. 2. Vyaas SP, Khur RK. Targeted and controlled drug delivery- novel carrier systems, 1st ed. Delhi: Cbs Publishers and Distributers; 2006. p. 414-458. 3. Jackson LS, Lee K. Microencapsulation and the food industry. 1st ed. Lebensmittelwissenschaft technologie 1991; 2;23. 4. Ming L, Olivier R, Denis P. Microencapsulation by solvent evaporation: state of the art for engineering approaches. Int J Pharm 2008;1:363. 5. Jain NK, Jain S, Bhandra D. Advances in controlled and novel drug delivery, 1st ed. Delhi: Cbs Publishers and Distributors; 2001. P. 426- 449. 6. Chien YW. Novel drug delivery system. 2nd ed. New York: Marcel Dekker Inc; 1992. P. 161-172 7. Julian BL, Leah EA, Scott MH, Thombre AG. Formulation design and pharmaceutical development of a novel controlled release form of azithromycin for single-dose therapy. Drug Dev And Ind Pharm 2009;35:1522-29. 8. The Indian Pharmacopoeia Commission, Ghaziabad. Indian Pharmacopoeia. 1996;2:246, 1:477- 78, 2:912- 913. 9. Mendham J, Denney RC, Barnes JD, Thomas MJK. Vogel’s textbook of quantitative chemical analysis. 6th ed , New Delhi: Pearson Education; 2006. p. 503 10. Beckett AH, Stenlake JB. Practical pharmacutical chemistry. 4th ed. Nodia: Cbs Publishers And Distributers; 2007; (Pt 2): P. 383, 275 11. Skoog DA, Holler FJ, Crouch SR, Instumental anaiysis. 2nd ed. New Delhi: Cengage Learning India Private Ltd; 2008. p. 343, 982. 12. Behera BC, Sahoo SK, Dhal S, Barik BB, Gupta BK. Chracterization of glipizideloaded polymethacrylate microspheres prepared by an emulsion solvent evaporation method. Trop J Pharm Research 2008;7(1):879-85 13. Deore BV, Mahajan HS, Deore UV. Development and characterization of sustained release microspheres by emulsion solvent diffusion method. Int J Pharm 2009;3:634-42. 14. Gao Y, Fu-De C, Guan Y. Preparation of roxithromycin-polymeric microspheres by the emulsion solvent diffusion method of taste masking. Int J Pharm 2006;318:62-9. 15. Hadkar UB. A handbook of practical physical pharmacy and physical pharmaceutics, 1st ed. Pune: Nirali Prakashan; 2004. p. 68-69. 16. Sinko PJ. Martin’s physical pharmacy and pharmuceutical sciences. 5th ed. New Delhi: Wolters Kluwer Health (India) Pvt Ltd; 2007. p. 4, 533, 540, 546, 553, 558. 17. Lachman L, Libermann HA, Kanig J. The theory and practice of industrial pharmacy, 3rd ed. Bombay: Varghese Publication; 1991. p. 412- 428. 18. Benita S, Benoit LP, Puisieux F, Thics C: Characterization of drug-loaded poly (d,llactidc) microspheres. J Pharm Sci 1984;73:1721. 19. Suhagia BN, Shah SA, Rathod IS. Determination of azithromycin in pharmaceutical dosage form. Int J Pharm 2006;68:242-45. 20. Abazinge M, Jackson T, Yang Q, OwusuAbabio G. Comparison of in-vitro and in-vivo release characteristics of sustained release drug delivery. Xenobiotica 2000;7 (2):77-81 21. Sultana N, Arayne MS, Hussain F. Degradation studies of azithromycin and its spectrophotometric determination in pharmaceutical dosage forms. Pak J Pharm 2006;19:94-98 22. United States Pharmacopoeial Convention, Rockville. Usp Nf 30. 2007; p. 2350. 23. Zhang ZY, Ping QN. Microencapsulation and characterization of tramadol- resin complexes. Int J Pharm 2000;66:107-113. 24. Patil.VB, Varsha BP. Preparation and evaluation of sustainedrelease nimsulide microspheres prepared from sodium alginate. Indian J.Pharm.Sci.2001:63(1);15-19. 25. Schimdt P. Secondary electron microscopy in pharmaceutical technology. In: Swarbrick J, Boylon, JC, editors. Encyclopedia of pharmaceutical technology. 3rd ed. New York:Marcel Dekker, Inc; 2007. p. 3217- 325. 26. Jens TC, Rhodes CT. Drug stability principles and practices. 3rd ed. New York:Informa Heathcare, 2005. p. 314-316, 322.
ABBREVIATION IR:
Infrared spectroscopy DSC: Differential Scanning Calorimetry XRD: X-Ray diffractometry SEM: Surface Morphology UV: Ultraviolet Visible double beam Spectrophotometer K: Rate constants R: Correlation coefficients AST: Accelerated stability studies
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesPERFORMANCE OF DFT FOR THE CALCULATION OF HYPERFINE COUPLING CONSTANTS OF SOME ISOTROPIC METAL COMPLEXES
English148153P.L.VermaEnglishVinita PrajapatiEnglish Dhirendra PrajapatiEnglishThe performance of DFT methodology to predict with accuracy the hyperfine coupling constants (hfccs) of isotropic metal complex, [Mn(CN)4N]- [Cr(CO)4]+,and Co (CO)4] have been investigated. For this investigation, , BHPW91, B3LYP, B3PW91, BLYP, BHLYP,BPW91, BP86, BHP86 functionals have been used. The hyperfine coupling constants values obtained from different density functionals have been found in the order of BHPW91> BHP86 > BHLYP > BLYP > BP86> BPW91> B3LYP > B3PW91 >EXP for [Mn(CN)4]2- , BLYP >BP86 >BPW91>B3LYP >B3PW91> BHLYP >BHP86 >BHPW91 >EXP for [Cr(CO)4]+ and BHPW91> BHP86> BHLYP> BLYP > BP86 > BPW91> B3PW91> B3LYP >EXP for [Co (CO)4]. The performance of the functional B3PW91 has been found better for Isotropic Metal Complexe [Mn(CN)4N]- ,BHPW91 for Isotropic Metal Complex [Cr(CO)4] and B3LYP for Isotropic Metal Complexe [Co (CO)4]. We have inferred that results obtained by DFT calculated EPR parameter hyperfine coupling constant values are in close agreement with experimentally observed values.
EnglishDensity Functional Theory (DFT). Isotropic Metal Complexes, Hyperfine coupling Constant.INTRODUCTION
Experimental method based on electron paramagnetic resonance phenomena belongs to most widely experimental techniques for investigation of molecular and electronic structure the difficulty with such experiments usually a proper interpretation of data obtained from high resolution spectra, opens new challenges for pure theoretical methods. One of these methods is density functional theory that now has an advanced position among the whole variety of computational techniques [11,12]. The computation of the structure of transition metal complexes has been an area of long standing interest, with clear problems not present in the corresponding studies of 1st and 2nd row species [5].Much of the growing acceptance by the physics and chemistry community of Density Functional Theory (DFT) is due to its success in treating transition metal complexes, in stark contrast to the failure of the Hartree Fock method. This technique presently the most successful approach to compute electronic structure of matter and hence for calculation of EPR parameters such as g-tensors and hyperfine coupling constants A of transition metal complexes [6, 22]. The basic idea behind DFT is to use the electron density rather than the quantum mechanical wave function to obtain information about atomic and molecular systems. While this idea came up in the very.
first years of quantum mechanics with the pioneering work of Thomas and Fermi in 1927[9] and was continued with Slater [24] the Hohenberg-Kohn theorems [7] are regarded as the real beginning of DFT. The most fundamental of these approaches originates from the pioneering work of Hartree and Fock [20] .Modern DFT rests on two theorems by Hohenberg and Kohn [8,10] .The major problem of DFT: the exact functionals for exchange and correlation are not known except for the free electron gas. However, many approximations exist which permit the calculation of molecular properties at various levels of accuracy. The most fundamental and simplest approximation is the local-density approximation (LDA), in which the energy depends only on the density at the point where the functionalis evaluated (Kohn and Sham) .LDA, which in essence assumes that the density corresponds to that of a homogeneous electron gas, proved to be an improvement over HF. While LDA remains a major workhorse in solid state physics, its success in chemistry is at best moderate due to its strong tendency for over binding. The first real breakthrough came with the creation of functionals belonging to the so-called generalized gradient approximation (GGA) that incorporates dependence not only on the electron density but also on its gradient, thus being able to better describe the inhomogeneous nature of molecular densities. GGA functionals such as BP86 [3]) or PBE [20] can be implemented efficiently and yield good results, particularly for structural parameters, but are often less accurate for other properties. The next major step in the development of DFT was the introduction of hybrid functionals, which mix GGA with exact Hartree–Fock exchange [2]. Now days, hybrid DFT with the use of the B3LYP functional [3] is the dominant choice for the treatment of transition metal containing molecules.This method has shown good performance for a truly wide variety of chemical systems and properties. Transition metal complexes have achieved considerable interest in the field of physics, chemistry, and biology. These metal complexes show a wide variety of biological activity such as antitumoral [13, fungicidal [1] bactericidal, or antiviral [14].They have been used for metal analysis for device application relative to telecommunications optical computing, storage and information processing. Several investigators have studied EPR parameters of transition metal complexes [4, 15, 16, 17, 18, 19, 21 23, 25, 26, 27, 28,] and have obtained good agreement with the experimental results. In this work we have represented the first extensive evaluation of DFT methods for the prediction of hyperfine coupling constants for [Mn(CN)4] 2- , [Cr(CO)4] + ,and Co (CO)4] Isotropic transition metal complexes. Results obtained for these complexes using eight different density functionals eight density functional BLYP, BP86, BPW91, B3LYP, B3PW91, BHLYP, BHP86, BHPW91 have been compared with reliable experimental data.
Figure1.0- Shows hyperfine coupling constant A values obtained from BLYP, BP86, BPW91, B3LYP, B3PW91, BHLYP ,BHP86, BHPW91, density functionals and experiment for Isotropic Metal Complex [Mn(CN)4N]-
Results - BHPW91> BHP86 > BHLYP > BLYP > BP86> BPW91> B3LYP > B3PW91 >EXP
Figure2.0- Shows hyperfine coupling constant A values obtained from BLYP, BP86, BPW91, B3LYP, B3PW91, BHLYP ,BHP86, BHPW91, density functionals and experiment for Isotropic Metal Complex [Cr(CO)4] +
Results - BLYP >BP86 >BPW91>B3LYP >B3PW91> BHLYP >BHP86 >BHPW91 >EXP
Figure3.0- Shows hyperfine coupling constant A values obtained from BLYP, BP86, BPW91, B3LYP, B3PW91, BHLYP ,BHP86, BHPW91, density functionals and experiment for Isotropic Metal Complex [Co (CO)4]
Results - BHPW91> BHP86> BHLYP> BLYP > BP86 > BPW91> B3PW91> B3LYP >EXP.
RESULTS
The results of the EPR parameter hyperfine coupling constant of different transition metal complexes considered for the DFT calculation in this work, obtained by the application of different density functionals BLYP, BP86 , BPW91, B3LYP , B3PW91 , BHLYP , BHP86 , BHPW91, and experiments are listed in Table No1.The hyperfine coupling constant values obtained from different DFT functionals of different complexes are also shown in Figure 1, 2,3.From the Figure 1, 2, 3, and Table 1, the results obtained by different density functional may be compared with experimental results. From the comparison of hyperfine coupling constants values obtained from different density functionals with experimentally observed values of the complexes considered in this study the performance of different functional have been found as follows. 1-Isotropic Metal Complex [Mn(CN)4N]- Results - BHPW91> BHP86 > BHLYP > BLYP > BP86> BPW91> B3LYP > B3PW91 >EXP. From these results it is inferred that result obtained by DFT functional B3PW91 is in close to the experimental result so the performance of this functional best for calculation EPR parameter hyperfine coupling constant A for Isotropic Metal Complex [Mn(CN)4N]- where as result obtained by BHPW91is very y poor and this functional is not appropriate for the calculation of EPR parameter hyperfine coupling constant A for this complex. 2-Isotropic Metal Complex [Cr(CO)4] + Results - BLYP >BP86 >BPW91>B3LYP >B3PW91> BHLYP >BHP86 >BHPW91 >EXP. In this case of Isotropic Metal Complex[Cr(CO)4] + , result obtained by DFT functional BHPW91 is in close to the experimental result so the performance of this functional best for calculation EPR parameter hyperfine coupling constant A for Isotropic Metal Complex Isotropic Metal Complex[Cr(CO)4] + where as result obtained by BLYP is very y poor and this functional is not appropriate for the calculation of EPR parameter hyperfine coupling constant A for this complex. 3-Isotropic Metal Complex [Co (CO)4] Results - BHPW91> BHP86> BHLYP> BLYP > BP86 > BPW91> B3PW91> B3LYP >EXP. In the case of Isotropic Metal Complex [Co (CO)4] , result obtained by DFT functional B3LYP is in close to the experimental result so the performance of this functional best for calculation EPR parameter hyperfine coupling constant A for Isotropic Metal Complex [Co (CO)4where as result obtained by BLYP is very y poor and this functional is not appropriate for the calculation of EPR parameter hyperfine coupling constant A for this complex. CONCLUSION From the above results it is inferred that that although the DFT calculated values of hyperfine coupling values obtained from different density functional for hyperfine coupling constant A have been found in close agreement with the experimental values but the better performance of the different functionals have been found different for different metal complexes .We have not identified any such functional, the results of which is excellent for all three metal complexes considered for study in this work. The performance of the DFT functional B3PW91 have been found better for Isotropic Metal Complex [Mn(CN)4N]- , B3LYP for Isotropic Isotropic Metal Complex [Co (CO)4] where as the DFT functional >BHPW91 is better for Isotropic Metal Complex [Cr(CO)4] + The theoretical results for hyperfine coupling constant obtained in this investigation suggesting that further development of density functionals is needed.
ACKNOWLEDGMENT
The authors acknowledge the immense help received from the research scholars and scientists 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 authors would like to thank Dr .R.N Patel ,Prof.K.B.Ponday and Dr.H.Surya Prakash.Rao for valuable suggestions.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareASSESSMENT OF DISABILITY IN TERMS OF ACTIVITIES OF DAILY LIVING (ADL) IN LEPROSY AFFECTED PEOPLE OF LEPROSY COLONIES AT BIJAPUR CITY, KARNATAKA
English154159Vinod S KambleEnglish Aparna Y TakpereEnglish Santosh M BiradarEnglishBackground: Disability in leprosy affects the activities of daily life which has got a negative impact on the quality of life. Further due to disability and social stigma there is restriction in participation in social functions. All these factors must be considered while measuring disability and rehabilitation. Objectives: 1) To assess the disability in persons suffering from leprosy and to find out the effect of disability on activities of daily living and social life. Design: A cross sectional descriptive type of study. Subjects: 132 people affected by leprosy, living in leprosy colonies at Bijapur, Karnataka, India were involved in the study. Methodology: A house to house survey and interview was carried out using the standard questionnaire of the Green Pasture’s assessment scale. Results: The prevalence of different types of impairments ranged from 31% in Lower Limbs, 35.9% in Upper Limbs and 45.46% in Eyes. The most commonly affected indoor activities were bathing (30.30%), washing hands and feet (30.30%), going to toilet (27.27%), dressing (31.82%), using scissors and knife (33%), eating and drinking (28.23%). Among the outdoor activities affected were cutting grass (51%), ploughing (61%) and carrying water pot (48%). Conclusion: Experiencing severe difficulties with the activities of daily living is a common problem in persons with chronic impairments due to leprosy. This leads to restrictions in social participation and sometimes even isolation as a result of disfigurement and social stigma. The level of difficulty can be assessed and measured using Green Pasture’s assessment scale based on subjective difficulty, which could be more helpful in rehabilitation.
Englishimpairment, social participation, rehabilitation, leprosy, Green Pasture’s assessment scale.INTRODUCTION
Leprosy is one of the major causes of preventable disability, including impairments, problems in activities of daily life and social exclusion resulting from stigma.1 Impairment and deformities (visible impairments) may cause limitations of activities of daily living (disability) and adverse social reactions (restriction of participation).2 Leprosy is still a major public health problem in several leprosy endemic countries with a worldwide prevalence of 212,802 patients at the beginning of 20083. In various studies done in India, the ratio between grade 2 and grade 1 disabilities ranged from 0.4-7.8.4 WHO disability grading does not grade disabilities but impairments. It is insensitive to change in the patient’s condition. 2 This leads to inappropriate way of rehabilitation and sometimes its failure, as it is based on impairment and not the subjective difficulties faced by the leprosy affected individuals. Therefore, a cross-sectional descriptive study was designed to find out the impairment and disability effect on the activities of daily living using Green Pasture’s assessment scale.5
Aims and Objectives
1. To assess the disability in persons suffering from Leprosy 2. To find out the effect of disability on activities of daily living and social life.
MATERIALS AND METHODS
A house to house survey and interview was carried out using the standard questionnaire of the Green Pasture’s assessment scale. 132 people were interviewed with informed verbal consent.
RESULTS AND DISCUSSION
As there is no well-defined concept of disability and there is confusion over terminology, less attention is paid in assessing disability.2 For rehabilitation the cause of the impairment that resulted in the disablement is less important than problems experienced by the person affected.2 Rehabilitation therefore aims at reducing disability and handicap.6,7,8 Hence this study was carried out to find the disability in terms of (ADL)Activities Of Daily Living using Green Pastures Assessment Scale which is based on subjective feelings or perception regarding disability. From the interview using the standard questionnaire the data obtained and analysed was as follows: From the interview it was found that maximum numbers of people affected with leprosy are in the age group 56-65 years. This may be because in the olden days, proper health infrastructure was not present, adequate man power was not available, and health awareness was not spread up to the grass-root level which is consistent with the study done by The Salsa Collaborative Study group.9 In the present study, 48.48% were males and 51.58% were females. In a similar study by Wim H van Brakel et al 74% were males and 26% were females.2 and by Fredrick J Slim et al 59% were males and 41% were females.4 It is seen that both the sexes have approximately similar prevalence rate in percentage suggesting that both sexes are equally prone to the disease. In our study, 84.85 %( 112) had taken MDT treatment. This finding is consistent with the study by Annamma Succhanda John et al where 90% patients had taken treatment for leprosy.10 This indicates that majority of people affected by leprosy have good compliance towards MDT treatment. The hurdles in good compliance are social stigma, tendency to hide disability and disfigurement, repeated hospital visits required for treatment, domestic responsibilities and cost or loss of daily wages due to hospital visit.11 Also women suffer more isolation and rejection than males.12 104 people (78.79%) said that their job/occupation was affected due to disability. In a study by F.J.Slim et al 23.8% people affected by leprosy said that their job/occupation was affected due to disability.2 82 people (62.12%) felt that they are physically and financially dependent on others. In study by Gopal et al 35% of the leprosy affected people and their family members faced social and economic problems. 13 In a study done by Nandgoankar Hemant et al 65.19% had problems in the work area due to disability.14 Hence activity limitation were found to be major determinants of participation restrictions and occupational loss which depends upon environmental, social and cultural variations along with gender discrimination.2,12This restriction affects the physical and financial dependency. Also certain occupations in leprosy affected may lead to deterioration of deformities causing further disability. 14 Due to muscle paralysis, ulceration, digital shortening, stiffness of fingers there is loss of skillful work and the affected people become jobless with no other alternative but begging because of social stigma. 15 94 people (71.21%) stay with the family members in the colony. 38 people (28.79%) are staying alone. In the study by Gupte M D et al, 8% of the leprosy affected people faced social isolation.16 In another study in India by Leprosy International Union, 43% of the leprosy affected people faced dehabilitation .17 To become dehabilitated is to get devalued, dispossessed of ones roles and functions in society and loose ones social identity. Social and financial factors are interlinked with each other and hence while planning for rehabilitation these 2 factors and the characteristics of the target population should be considered. 15 RELATIONSHIP: Relationship with family members and others with some and many problems was as under:
a) It was noticed that Relationship with the spouse and children was affected in the range of 7-16.66%. In the study by Ms. W. Chitra the percentage was between 6-24%.18 A study by International Leprosy Union showed that 43% of the total 1071 leprosy affected people faced dehabilitation and had to leave their homes.17In a study done in Orissa it was found that 45.90%(308) of families with leprosy patient faced social isolation in which 48.3%( 150) had deformity and 51.30%(158) had no deformity.19 The various reasons for social isolation are 1) fear of infection 2) denial and rejection by the community members due to stigma. 3) Stigmatizing and isolating oneselves.20 Hence along with physical treatment and rehabilitation, social, vocational and mental rehabilitation is very much required. 19.69%(9) people found some difficulty in the use of assistive devices and 9.09% had much difficulty while the study by Ms.W. Chitra showed that 66% found some difficulty and 22% found more difficulty in use of assistive devices.18 69.69% people said assistive devices were not necessary and 28.28% said there is a need of the devices. This suggests that the use of assistive devices to help in activities and participation in life areas such as work and employment is not utilized on a broad scale. 11This is because of lack of knowledge and awareness of assistive devices, non-provision of the assistive devices by the health services and underestimation of disability with underutilization of the assistive devices for hiding the disability due to stigma by the leprosy affected people themselves. In the present study, 90.90% people were unemployed and 9.10% were employed. Out of the employed 41.66% were employed daily and 58.34% were employed sometimes. In present study only 4 people were farmers and 8 people were laborers. 90.91% (120) people had no jobs and so they did begging for survival. In the study by Wim H. van Brakel et al. 63% were farmers, 9.7% were working in the house, 6.3% were students, only 0.7% were beggars and 145 were doing some other work.2 All the people affected by leprosy face difficulty in their jobs. For the less disabled who still could work in the field, life was easier than for those who were older and with severe disability. Due to severe disability, disfigurement and social stigma most of these leprosy affected people have no other alternative but to succumb to begging for the livelihood. Hence along with physical rehabilitation, vocational and financial rehabilitation is very much required.
Impairment: In our study impairment in vision was 45.46%, for lower limbs it was 31% and upper limbs was 35.9%. In a study by Nandgoankar Hemant et al, the percentage of involvement of eyes, upper limbs and lower limbs was 5% , 79% and 63%, which is high compared to the present study.14 and by Fredrik J Slim et al it was reported to be 41%, 68% and 82% which is also high.4 Here in the study, upper and lower limb impairment was low in percentage and vision impairment is similar when compared with other studies above. Impairment of the eyes, hands and feet leads to severe disability while performing activities of daily living and social responsibilities which in turn changes to social handicap. Though the measurement of impairment helps the clinician to plan for physical and medical rehabilitation, the measurement of impairment do not give the true picture of disability as disability is contextdependent and subjective. One individual with a given impairment may experience much more difficulty with certain activities than someone with the same impairment living under different conditions. Hence rehabilitation requires us to look at the situation from the patient’s point of view.2
CONCLUSIONS
Facing severe difficulties with the activities of daily living is a common problem in people affected with leprosy related impairments and the level of difficulty can be assessed and measured using Green Pastures Assessment Scale. People affected by leprosy have problems in social participation as a result of disability, disfigurement and social stigma. Disability in activities of daily life has a negative impact on the quality of life. The relations amongst the family members and others are also stressed. Jobs are affected leading to unemployment, leaving the leprosy affected people no other option for survival but begging because of insufficient government schemes. All these factors must be considered while measuring disability and planning for rehabilitation. Recommendations: It is recommended that disability assessment should be a standard activity for planning, monitoring and evaluation of rehabilitation, both for individuals and at various rehabilitation programs. Knowledge of the disability status of a person will be valuable in needs-assessment for rehabilitation interventions and in clinical decision making.
ACKNOWLEDGMENT
The authors wish to acknowledge all the staff who was involved in the study. Also we wish to thank the people affected by leprosy living at leprosy colonies for their cooperation and participation in the study. The immense help received from the scholars whose articles are cited and included in references of this manuscript cannot be forgotten. 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=2010http://ijcrr.com/article_html.php?did=20101. Wim H Brakel –Disability And Leprosy: The Way Forward-Annals Academy Of Medicine Jan 2007, VOL.36 No 1:86-87
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4. Frederik J Slim,MD, Carine H.van Schie ,PhD, Renske Keukenkamp, MSc, William R. Faber, MD, PhD and Frans Nollet ,MD, PhD. Effects of impairments on activities and participations in people affected by Leprosy in Netherlands. J. Rehab Med. 2010; 42: 536- 543)
5. W.H.VanBrakel,A.M.Anderson,F.C.Worpel,R .Saiju,H.B.BK,S.Sherpa,S.K.Sunwar,J.Gurung ,M.Deboer and E.Scholten A scale to assess Activities of Daily Living in persons affected by leprosy. Asia Pacific Disability rehabilitation Journal,1998; vol.9,no 1, 1-12
6. Van BennekomC A M,Jelles F,Lankhorst G J. Rehabilitation activities profile:The ICIDH as a framework for a problem-oriented assessment method in rehabilitation medicine.Disabil.Rehabil.1995;17:169-175.
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10. Annamma Succhanda John,Pamidipani Samuel Sunder Rao and Sonali Das. Assessment of needs and quality care issues of women with leprosy.Lepr Rev.2010;81:34-40.
11. Johan Borg and Stig Larsson. Assistive devices for people affected by leprosy: underutilized facilitators of functioning? .Lepr Rev.2009 Mar;80(1):13-21.
12. Sanjay P. Zodpey, Rajnarayan R. Tiwari and Atul D. Salodkar. Gender differentials in the social and family life of leprosy patients. Lepr Rev.2000; 71: 505-510
13. Gopal P K. Methods to identify the Leprosy Patient Needing Rehabilitation. Indian Journal of Leprosy1997; 69:133-152
14. Nandgaonkar Hemant P,Mancheril Joy,Ebenezer.J,Samy A.A. Activities of Daily Living (ADL) Assessment :A measure for grading activity limitation in Leprosy patients . The Indian Journal of Occupational Therapy 2003; vol34 (3): 8-12.
15. H Srinivasan .Developmental Articles: The problems and challenges of disability and rehabilitation in leprosy. Asia Pacific Disability Rehabilitation Journal1998.Vol.9;No.1
16. Gupte M D et al.-Personal Communication 17. International Leprosy Union .A Study of social Aspects of Dehabilitation in Leprosy, Pune, 1994.
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19. 15.Srinivasan H and Sivakumar S S.Personal Communication.
20. Chen, Tongsgeng Chu and Qihua Wang. Qualitative assessment of social, economic and medical needs for ex-leprosy patients living in leprosy villages in Shandong Province, The People’s Republic of China ,Shumin. Lepr Rev 2005; 76: 335–347
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29General SciencesCUSTOMER ACCEPTANCE OF eBANKING: CUSTOMIZED MODEL FOR eBANKING PRACTICES IN PAKISTAN
English160170Muhammad SiddiqueEnglish Allah Nawaz English NajeebullahEnglishComputer-based banking (e and iBanking) is mushrooming around the globe. Both advanced and developing countries are making all out efforts to adopt digital technologies and thereby receive the benefits for their masses in general and banking sector in particular. However, experiences of different countries in taking digital opportunity initiatives (DOI) for this purpose reveal that uptake of information and communication technologies (ICT) is neither automatic nor absolutely technical rather a „social-process? where several factors play significant role in determining user-acceptance of technologies. This paper develops a customized theoretical model of the issue from the existing research that will be tested empirically.
EnglisheBanking, iBanking, ICT, DOI.INTRODUCTION
The application of electronic communication in business dates back than the 1970s. About 1918, the payments between banks were done electronically through telegraph. This use has gradually inflated with the passage of time and currently almost all the payments between banks and other organizations are done electronically (Singh et al., 2002; Siam, 2006). Today, a click of digital button offers the customers with eBanking services by empowering them with extraordinary freedom in choosing vendors for their financial services. The states of the world have no other option than to adopt eBanking because of globalization and high competition (Wahab et al., 2009; Nawaz, 2010; Banan, 2010). Several studies show that competitive pressure is the principal driving force behind taking digital opportunity initiatives (DOI) for iBanking, ranking ahead of revenue enhancement and cost reduction. By using the internet technology, banks offer iBanking services which serve the needs of the current customers and magnetize the new ones (Comptroller, 1999:3). Financial institutions have removed the borders between themselves, and introduced innovative products and services and made available the current ones into several packages (Siam, 2006). However successful adoption and use of IS, is vital for the growth and development of financial institutions (Kuppusamy et al., 2009; Wahab et al., 2009). In contemporary digital banking the customer fidelity can change instantaneously by clicking the computer-button only, therefore, all the banks have to recognize the importance of different factors responsible for the success and failure of modern banking. They must study the role of these factors in the current business environment, so as to manage them successfully to remain in competition in the rapidly changing world (Singh et al., 2002). In addition to these, the banks should attract and stimulate people through extensive advertisement efforts and incentives of cost cutbacks and other ones. According to TAM, Perceived usefulness of customer about the new technology is essential. Each society that wants to reach its goals in implementing eBanking should pay attention to these issues vigilantly and have a complete program and sufficient investment in this way (Salehi and Alipour, 2010).
E-BANKING: CONCEPTS and PRACTICES
Literature defines eBanking as web-based-banking and it has stayed a high-risk area from the very start of its innovation, for example; in 2001 about $17 billion was spent on the security of information-related products and services in America (Hertzum et al., 2004). Online banking is based on the Internet gateway, which provides different types of online banking services like payment of utility bills, buying the mobile recharge and investing online. It is thus, necessary for the banks to establish their official websites providing the option for online business (Pikkarainen et al., 2004; Salehi and Alipour, 2010). In the eBanking powers is delegated to the customers for self-service by satisfying certain banking needs and thereby ease themselves in a number of ways: like they can examine their account histories, pay bills and transfer funds, activate or restore credit card (Amor, 1999). eBanking offers the facility to the customers to access their accounts and carry out transactions in a trouble-free way by visiting the websites of the bank. All the individuals and companies are cutting-back a lot of their time and cost because online banking is proving less costly as compared to the traditional banking transactions (Karjaluoto et al., 2002, p.261). Internet offers a podium for using online banking services from inside and outside the organizations thus, ICT-adoption is steadily transforming businesses from local to global (Polatoglu and Ekin, 2001). According to Giglio (2002) for delivering banking products and services the cheapest delivery channel can be online banking. With the help of online banking services, the branch networks of banks have condensed and the banking staff as well as the customers are pleased with online banking services as it saves time and effort to carry out transactions (Karjaluoto et al. (2002).
E-BANKING IN PAKISTAN
The development and growth of ICTs is quiet slow in Pakistan as compared to other developing countries. Here, the ePolicies focus to develop the domestic ICT sector for the development of eCommerce in the country. For this purpose, the government of Pakistan has established seven IT universities and one virtual university under the National Information Technology Policy to produce qualified professionals to meet the existing needs of digital society (Khan and Bawden, 2005). Similarly, the National Education Policy (1998-2010) seeks to upgrade the education level in Pakistan by offering IT courses at different levels of education to ensure digital literacy among the youths. Some renowned IT companies of the world like Microsoft, Cisco, and Oracle are also making huge investments in Pakistan for the promotion of ICTs (Shahzada, 2006; Pasquet et al., 2008). eBanking is becoming attractive to both the banks and the customers because this new technology is increasingly acceptable to them. Similarly, banks are also facing huge competition in the market and they can increase their market shares by offering high quality services to the customers. The ATMs and credit card services were introduced in Pakistan by City Bank in mid 1990s, which was then followed by the domestic banks in the late 1990s (Khan and Bawden, 2005). eBanking is the use of ICTs and other electronic means by a bank to carry out transactions and to maintain interaction with all the stakeholders (Abid and Noreen, 2006; Hasan et al., 2010). The Government of Pakistan started digital initiatives in early 2000. The banks have got the lead in eCommerce but most of the progress has been made in eGovernment. Some business to business gateways are available which are designed more for information rather than transactions. The de-regulation policy of the country has encouraged many private and foreign banks to establish strong end user by adopting eCommerce (Ahmed, 2006; Hamzaee and Hughs, 2006). The Ministry of Science and Technology has taken several steps to promote eCommerce and eBanking in the country. In Pakistan, high technological flow has forced essential changes in the financial industry; new business plans have emerged and opened ways for doing business. eBusiness has launched the use of IT for improved internal controls and more complex risk management systems, and this in turn has resulted in improved, high quality, and convenient customer services (Akhtar, 2006).
CUSTOMER ACCEPTANCE
Models (Theories) of Customer Acceptance
It is widely established that customer acceptance is the prime factor in determining the rate of change in the financial sectors (Sathye, 1999; Floh and Treiblmaier, 2006; Abukhzam and Lee, 2010). A number of models have been suggested to examine, recognize and apply strategies for creating user acceptance of new technologies. The most commonly used research models include: TAM (Davis, 1986), TRA (Fishbein and Ajzen, 1975), TRI (Parasuraman, 2000), DI (Rogers, 1962), and TPB (Ajzen, 1985, 1991). These models have been used by several researchers to discover the dynamics of „user acceptance? of eBanking (see for example, Kuppusamy, et al., 2009; Andoh-Baidoo, and Osatuyi, 2009; Amin and Ramayah, 2010). a. Technology Acceptance Model (TAM) TAM was devised to identify a small number of basic factors suggested by earlier research and dealing with the cognitive and affective determinants of computer acceptance (Sathye, 1999). TAM defines the two constructs, that is, perceived usefulness (PU) and perceived ease of use (PEOU) that are of principal importance for computer acceptance. Perceived usefulness (PU) is defined as the level that using a specific system will increase the job performance while Perceived ease of use (PEOU) refers to the extent to which the user considers that the target system is to be free from effort (Davis et al., 1989). TAM was emerged from the theory of reasoned action (TRA) and proved to be the generally accepted model (Jahangir and Begum, 2008). b. Technology Readiness Index (TRI) TRI (Parasuraman, 2000) refers to people?s inclination to hold and use new technologies for the achievement of their goals. The TRI includes four factors: optimism, innovativeness, discomfort, insecurity. Optimism means to what extent the people have positive view of technology for improving their life standards. Innovativeness refers to the level of people for becoming pioneers in technology adoption and hence, the thought leaders. Discomfort means the extent to which people perceive a lack of control over technology and feel overwhelmed by it; and insecurity: is the degree to which people have doubts about technology and are uncertain of its capacity to work properly (Gerrard et al., 2006). c. The Theory of Reasoned Action (TRA) This theory was developed by Fishbein and Ajzen (1975) and improved afterwards by experimental facts to hold up its strength and consistency. It was assumed that an individual?s behavioral intention is the instant determinant of behavior, his/her attitude and subjective norms are mediated through behavioral intention and their behavioral and normative beliefs are mediated through attitude and subjective norm (Ajzen andFishbein 1980). Subjective norm is beliefs about what others will consider about the behavior; in other words, the perceived influences of social pressure on an individual to perform or not to perform the behavior (Sadeghi and Farokhian, 2011). d. The Theory of Planned Behaviour (TPB) After recognizing some problems with the Theory of Reasoned Action, it was designed specifically a modified model: Theory of Planned Behavior (TBP) to predict and explain behavior based assumption “person?s volitional control” to influence the adoption of new technology (Ajzen and Madden 1986:457). TRA was expanded by adding another construct called Perceived Behavioral Control (PBC), which refers to an individual?s perception of the presence or absence of required resources and opportunities to carry out the specific behavior (Ajzen, 1991). The TPB presume that “the behavior is determined by the intention to perform the behavior (Benham and Raymond, 1996)” and this intention is determined by three factors: attitude, subjective norms and perceived behavioral control. Each factor consists of a number of beliefs and related evaluations (Mashadi et al., 2007). Plenty of experiential proof proposes the TPB capacity to effectively explain one?s intentions and behavior in accepting new information technologies. TAM based on Theory of Reasoned Action (TRA), and afterwards was developed to be the Theory of Planned Behavior (TPB) (Kasemsan and Hunngam, 2011). e. The Diffusion of Innovations (DI) The theory of DI (Rogers, 1995) explains diffusion of innovations as: “… the process by which an innovation is communicated through certain channels over time among the members of social systems. It is a special type of communication, in that the messages are concerned with new ideas”. A decision to adopt an improvement purely depends on the perceived characteristics of innovations, i.e., compatibility, relative advantage, trialability, and observability are to be considered (Al-Hajri, 2008). The increased use of integration of services around digital networks (ISDN) and electronic data interchange (EDI) protocols are fundamental in the new distribution channels such as smart cards, telephone transfer systems (TTS), and electronic fund transfer at point of sale terminals (EFTPOS). Card technology such as VISA and Master Card International Networks is evolving to offer customers with border-free services (Kuppusamy et al., 2009). The modern distribution channels permit banks to offer more services and consequently have great effects in the banks? cost structures (Kasemsan and Hunngam, 2011).
Factors of Customer Acceptance
Issue-Related Variables
a. Government ePolicies (GEP)
The IT policy (ePolicy) plays dominant role in determining the prospects of eBanking in any country including Pakistan (Zarmeene, 2006). Consistent policies with due support of the government helps in the promotion of IT culture in the country and stimulates eBusiness. Pakistan is confronted with numerous challenges in IT adoption (Kundi and Shah, 2009). Similarly, legal support has a huge impact on users? iBanking acceptance as responsibility must be fixed when financial losses happen in Internet transactions (Kasemsan and Hunngam, 2011).
b. Quality of Internet (QOI)
eBanking uses the web browser for the user interface and the Internet for data transfer and software download, and so has an advantage of cost-reduction and speedy transmission of information (Hertzum et al., 2004). From a technological and cost-driven perspective it may appear quite logical for the banks to switch over to online however, at the same time the problem of how to foster customer loyalty arises when the connection between the bank and the user becomes virtual (Floh and Treiblmaier, 2006). Thus, the overall quality of eBanking has captured greater attention of bank managers and researchers due to its sturdy impact on customer loyalty, customer satisfaction, costs, profitability, and business performance (Sadeghi and Farokhian, 2011).
c. eBanking Awareness (EBA)
The researchers tell that there is a significant statistical relationship between awareness, access to Internet facility, length of banking relationship, people working in the iBanking, education level and the income level with the usage of iBanking (Padachi et al., 2007). A research tells that the main reason why customers are unwilling to use iBanking is information diffusion between the banks and their customers. This problem was articulated this way “the youth are more likely to use iBanking than the older citizens because they are more familiar with the Internet but for older customers, there is a need to entice them through Internet awareness programs” (Andoh-Baidoo, and Osatuyi, 2009).
d. Perceived Usefulness (PU)
It is natural that if a human being considers a technology valuable, he/she gets mentally prepared to learn about even by taking pains. The user resistance to change is reduced due to the positive effects of the modern digital devices. Perceived usefulness is the extent to which an individual considers that using a particular system would improve his performance (Al-Hajri, 2008). It is also correct that perceived usefulness depends completely on the level of customer awareness about the features and functions of new technologies. If they have inadequate information about the utilities provided by the new devices, it is very much possible that users may misjudge the usefulness of eBanking (Riyadh et al., 2009).
e. Perceived Ease of Use (PEU)
It is commonly acknowledged that user acceptance is radically related with the „perceived ease of use and usefulness of modern technologies. If users value them negatively, their attitude towards change will also be negatively prejudiced. Moutinho and Smith (2000) studied the behavior of bank customers and concluded that simplicity of baking operations and convenience are the two important expectations. Furthermore, the variety of eBanking users and the lack of any special training to ensure ease of use are the important concerns to successfully implement the eBanking strategies (Hertzum et al., 2004). From the viewpoint of technology, ease of use is usually regarded as an important quality characteristic in computer services (Floh and Treiblmaier, 2006; Shih, 2007; Al-Hajri, 2008; Riyadh et al., 2009; Amin and Ramayah, 2010; Adesina and Ayo, 2010).
g. Security and Privacy (SandP)
People understand about the risks but they have pessimistic view regarding protection from ibanking risk. It has been found that customers have confidence on the bank but not sure about the security and privacy of their confidential information (Roboff and Charles, 1998). The researchers have also found that though the customers? confidence on the bank was strong but they were having weak confidence on the technology (Howcroft et al., 2002). eBanking must be safe, secure and easy to use. Automation, instruction, and understanding can be recognized as three approaches to practical security. Instruction is the main approach of the systems evaluated; automation ease the user from taking part in security, at a possible level; and understanding goes beyond step-by-step commands, to permit users to act proficiently and securely (Hertzum et al., 2004).
h. Trust of the Customer (TOC)
In order to examine the significance of eLoyalty, the identification of factors influencing recurring purchasing behavior and word-of-mouth suggestion is a crucial area of research. The extensive adoption of online banking services calls for research to investigate the factors which determine eBanking customers? loyalty. The most imperative factors affecting eLoyalty are unavailability of infrastructure, lack of trust on technology, service charges and security and privacy (Padachi et al., 2007). In response to these demands banks attempt to enhance the satisfaction of customers by offering improved products and services at reduced operating costs. Thus, customer trust plays a central role for eReadiness to use eServices (Wahab et al., 2009).
i. Quality of eBanking Services (QOS)
The internet infrastructure plays a very important role in eBanking and some grave steps should be taken for the development of internet. If we compare the current infrastructure with the five years before, we observe that a great many technological advancements have been made especially in the private sector and the credit of all this goes to the Government of Pakistan for fostering the privatization of banks in Pakistan (Pauline, 2001). This has resulted in the development and increase of IT infrastructure, online branches, ATM machines, ePayment systems, and eTransactions (State Bank of Pakistan, 2003).
Demographic Factors
Customer demographics play an important role in shaping their behaviors towards new technologies. There is a series of research studies on the measurement of demographic implications on the users of computer based information systems including eBanking (see for example, Ramayah et al., 2003; Shih, 2007; Padachi et al., 2007; Yang and Ahmad, 2009; Adesina and Ayo, 2010; Amin and Ramayah, 2010). Demographics have been identified as the intervening and/or moderating variables in affecting the relationships between the determinants of customer acceptance in case of using eBanking (Tat et al., 2008; Wahab et al., 2009). Similarly the researchers have explored and tested several demographic characteristics however some of these are very frequently used in researches on eBanking. These characteristics include: Gender, education, marital status, position (Ramayah et al., 2003); Gender, age, involvement, seeking behavior and technophobia (Floh and Treiblmaier, 2006); age, education, and income (Padachi et al., (2007); experience with eBanking, experience of using Internet, and frequency of use at least once a week (Shih, 2007); Gender, marital status, education level, and religion (Amin and Ramayah, 2010). In this research we have used the following demographics for analysis: Education, Gender, Age, Experience with eBanking, eBank(s) used, and Frequency of Use.
Factors of Customer Acceptance
From the literature review (presented above) a customized conceptual model (Figure 1) has been constructed which is grounded in the literature on one hand and reflects the current environment in Pakistan?s eBanking and iBanking industry on the other hand. The hypotheses emerging from the model have been identified which will be tested in the field surveys to be conducted by the researchers in near future.
DISCUSSIONS
All businesses are the recipients of eBanking, irrespective of their size or physical locations. Internet has made it possible to perform all types of commercial transactions on an electronic medium without geographical restrictions (Yang and Ahmed, 2009). The banks have been proved as the principal user of ICTs for many years because the use of IS helps in cost reduction, mainly due to the reduction of bank branches and their staff. The understandings and control of operational risks has also become easier for the banks using the modern Information Systems. Moreover, the successful use of Information System enable banks to process credit card and loan applications speedily, without wasting time in too many formalities (Wahab et al., 2009). There is a massive gap between developed and developing countries in terms of development and growth of eBanking services. The developed countries are far ahead in the adoption and use of leading-edge technologies in business and commerce, whereas the developing countries are mainly lacking the modern technology infrastructure, and where it is available, a large number of people are not familiar with the use of technology and thus, not using it (Pasquet et al., 2008). The banking industry has made substantial progress and the use of Internet technology has turned out to be a strong force changing the very core of conventional banking. So, technology is no doubt a single most leading strategic issue that has created regulatory challenges for eBanking (Banan, 2010). Investments of an organization in IS creates different problems, such as hardware and software inappropriateness, information overload, and feelings of job insecurity among staff members who fear that they will be replaced by machines. These problems may have unfavorable effects on productivity as well as lessen the growth and use of other IS applications (Kuppusamy et al., 2009). Least-developed countries are facing a shortage of quality IT professionals to design, develop, and operate eBanking systems. A short term solution to this problem is to launch, on emergency basis, the high quality training programs for IT professionals to foster the development of eBanking applications, but the development of modern high quality higher education institutions, to promote the educational standard of masses would be a real long-term solution (Hasan et al., 2010).
CONCLUSION
Currently the customers in the developed countries like UK, USA, Sweden, and Denmark are enjoying the benefits of eBanking but Pakistan is still lagging far behind due to several factors (Shahzada, 2006). For example, in Pakistan the acquisitions, installation, use, and maintenance of eBanking technologies along with user training are big issues. Similarly, due to below standard and poorly established IT education institutions, the developing countries are facing serious shortages of qualified IT professionals to meet their national requirements of eBusiness (Kundi and Shah, 2009). When asked about the key challenges and complexities of existing eBanking operations, fifty percent people argued for the lack of in-house IT professionals. The small banks have outsourced the development of their website design and eBanking service operations but they suffered from delays and slow responses of outside IT professional (Yang et al., 2005; Hamzaee and Hughs, 2006). IS-savvy customers refer to the customers? understanding and acceptance of eBanking services. Although the Banks? web traffic statistics indicate rise in the number of web visitors due to anytime, anywhere convenience of eBanking, but this increase in visitor traffic is not matched by a related increase in iTransactions. Regardless of the increase in online visitors and use of the website, just a mere tenth of households have used Internet for their financial transactions (Kuppusamy et al., 2009). This is due to the fact that today?s world has not only made remarkable developments in every field and has created exciting opportunities for the people and organizations, but has also caused general uncertainty, which is often a great concern of eBanking users (Banan, 2010). Today in a highly competitive economy, banks are investing huge sums of money on ICTs to meet the growing demands for eBanking and they are also well are of the fact that failure to do so will lead to their demise (Kuppusamy et al., 2009). Banking sector has achieved substantial growth and the Internet technology has turned out to be a strong force shifting the pattern of banking from conventional banking to eBanking. Therefore it can be claimed that technology is the single most leading strategic weapon that has created paramount challenges for the banking sector (Banan, 2010).
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareGASTROSCHSIS ASSOCIATED WITH OTHER ANOMALIES - A CASE REPORT
English171175Arun Kumar.S.BilodiEnglish M.R.GangadharEnglishAim of the study :-The objective of present study is to report a case of gastrochisis associated with multiple congenital anomalies that is born to consanguineous mother Place of study : in the labor ward at Teaching Hospital of Raja Rajeswari Medical College,Bangalore, Period of Study:-During three years period of study (2008—2010) Case Study :-A male child was born to consanguoious mother in the labour ward of above institution with presence of coils of small intestines outside the body wall associated with jejunal atresia ,imperforate anus and club foot. The Scoliosis of spine (deformity of spine).was also present and other systems were normal. No other anomalies were found in this new born male baby. Discussion: Since this is a case of Multiple Congenital Anomaly (MCA), this case has been well compared,,correlated and discussed with the available literature of other cases showing multiple congenital anomalies showing gastrointestinal Anomalies like gastrochisis , amphalocele,atresia of any part of gastrintestinal tract,imperforate anus etc. Conclusion:This is case of Gastroschisis associated with Multiple Congenital Anomalies (MCA) has not only profound embryological importance but also give profound knowledge of gastroscisis with multiple congenital anomalies of gastrointestinal tract to the Anatomists, Surgeons and Physicians . Hence this case has been studied well and reported
EnglishGastroschsis,-- Abdominal wall defect .Paraomphalocele -Abdominoschisis—LaproschisisINTRODUCTION
Gastroschsis is defect in the abdominal wall in the developing fetus during pregnancy by the formation of a hole in the abdominal wall through which allows intestine and other organs come out. These are not covered by any protective sac but exposed to amniotic fluid. Intestines may be shortened,twisted,swollen or may become irritated .Child may require surgery immediately after birth by pushing back the intestinal coils into the abdominal cavity Even after surgery child may have problem of feeding, digestion and absorption of food.”.According to the .Center for disease control and prevention(CDC), around 1871 babies are born with gastrochisis every year in USA1 . It has been stated by some authors that Gastroschsis occurs to the right of umbilicus and may be associated with Artesia of intestines (10%) .(Cincinnati children’s Hospital Medical center.). Maternal History This anomalous child was born normally to consanguoious mother after term in the middle class family..This was third child to her and other two children (one male and a female child )were normal but they were born by caesarean sections There was history of hydramnios during this pregnancy and family history of consanguinity, but there was no history of taking drugs or infection during first trimester. No history of smoking and alcohol intake. No systemic diseases in the family. METHODOLOGY A live male child was born to consanguoious mother aged 23 years in the labor ward at Teaching Hospital of Raja Rajeswari Medical College,Bangalore.. Once the child was born, cried well , minimum muconium was present on the body .Baby was cleaned well and weighed. The weight of the .new born male child was 3.0.kg..There was no difficulty in respiration. All the systems were examined in detail and they were found to be normal except Gastro intestinal system and anterior abdominal wall.. On general exaination Head and neck – Normal,, Face – normal, Chest – normal Extremities :-Upper limbs were normal There were ten digits. with no anomalies of digits . Spine : Scoliosis of spine (deformity of spine).was present Lower limbs:-Normal but there was club foot(deformity of foot) of the right foot .left lower limb was normal with normal ten digits No polydactyly or syndactyly of both upper and lower limbs were found in this new born child. Local examination of the abdomen On the detailed examination, child had defect in the anterior abdominal wall. with coils of intestine lying outside the abdominal wall These were coils of small intestine were not covered by any protective membrane or sac There was part of stomach herniating The opening of aperture was quite large measuring approximately 5.5 cms . There was also associated anomalies like jujenal atresia ,imperforate anus. Scrotum was normal .Testes were present in the srcotum.No other congenital inguinal or umbilical hernias were found. No anomaly of bladder or other pelvic organs..No other anomalies observed in this new born baby other than above anomalies. After examination was complete, photograph of anterior abdominal was taken.
DISCUSSION
The incidence of Gastroschsis in United States is 1 in 5000 infants. Cases of Gastroschsis are increasing world wide and in United States of America. This congenital disorder also has affected in infants having background of ethnic. These anomalies can be diagnosed by ultra sound also but exact etiology is not known in the fetus About 5 cms vertical opening is present in the abdominal wall adjacent to the umbilical cord Rarely large intestine along with other organs may be herniated through the opening New borns with gastrochisis are either have a low birth weight or prematurely born. They have associated anomalies like bowels which are underdeveloped or part of genetic syndrome or disorder Pregnant mothers have shown increase level of alpha feto protein in their blood due to presence of gastrochisis of fetus Mothers using recreational drugs in early period of pregnancy or smoke during pregnancy are highly prone for getting infants with Gastroschsis. 2 . Mothers of younger age are likely to have babies with Gastroschsis and mothers of white teenagers have higher incidence of Gastroschsis infants than in Black or African –American teenagers3 Babies with Gastroschsis are found more in mothers who consume more of alcohol and use tobacco in the form of smoking4 Use of certain drugs like Ibuprofen during pregnancy carries high risk of delivering babies with gastrochisis and also in women who have repeated urinary tract infections before pregnancy or early part of pregnancy have high risk of delivering babies with Gastroschsis5 Yuvaraj Bhosale et al (2007) have reported a case of multiple congenital anomalies having Omphalocele,extrophy of cloaca with imperforate anus and,Spinal defects(OEISComplex) in a still birth full time fetus to non consanguineous mother of 21 years old.There was no history of drug intake nor infection in her first trimester. Antenatal ultra sonography revealed omphalocele associated with polyhydramnios, congenital dislocation of hip on both sides with calcaneo vulgus feet deformity ,imperforate anus,genital ridge was unfused and there was splitting of genital tubercle .Hence sex could not made out . There was also kyphosis of lumbar spine.6 In India Incidence of multiple congenital anomalies(MCA) is 1.94% - 2.03% as per the analysis of all published studies 7and8 According to I.C.Verma (1978), the commonly occurring multiple congenital anomalies are defects of neural tubes with anomalies of cardio vascular systems, or musculo-skeletal system ,and gastro intestinal systems9The causative factors of major multiple congenital anomaly is due to genetic factors (30%-40%)environmental factors (5%-10%).Among anomalies caused by genetic, chromosomal abnormality constitutes only 6% , disorders of single gene constitutes 25% and multifactorial constitutes 20-30% and cause of 50% of multiple congenital anomalies is idiopathic 10. A study was conducted by Saarah Waller et al from 1987 to 2006 on all cases of live born infants with gastroscisis.They established a link between infants with gastroscisis and agricultural chemicals like atrazine ,nitrates,and 2,4dichlorophenoxyacetic acid .They found occurrences of gastroscisis was high due to contamination of water with atrazine Incidence of gastroscisis risk was also increased in a women who concieved in the spring that is between month of March,April and May due to more use of chemicals.during these months11 . This is case of multiple congenital anomalies seen in full term new born male baby involving gastro intestinal system(showing gastroscisis, jejunal atresia imperforate anus,),skeletal system club foot(deformity of foot) and scoliosis of spine, (deformity of spine) and polyhydramnios, but there was no congenital dislocation of hip on both sides nor omphalocele as seen in studies of Yuvaraj Bhosale et al (2007) studies No incidences of neural tube defects, nor anomalies of cardiovascular systems unlike in studies of I.C.Verma (1978), PRESENT STUDY In present case, a full term male baby was born normally to 23 years old mother of Bangalore in labor ward at Teaching Hospital of Raja Rajeshwari Medical College, Bangalore, Karnataka with coils of intestine lying outside the Anterior Abdominal wall. These coils of small intestine were not covered by any membrane or sac. On close examination, there was part of stomach also herniating along with coils of intestine on close examination The opening of aperture was quite large(approximately 5.5cms wide),In this case, environamental factor has not played any role unlike in studies conducted by Sarah Waller et al On examination of abdomen Coils of intestine were moist with adequate blood supply,and there was atresia of jejunum. Stomach was just protruding. There were no other organs seen at opening of the anterior abdominal wall.There was associated anomaly of club foot(deformity of foot) and scoliosis of spine (deformity of spine). No anomaly of bladder and other pelvic organs It is second degree consanguinity, hydramnios, and past family history of anomalies which may be the contributing factors for the occurrence of the present anomaly of Gastroschsis. The immediate surgery was done on this child by placing all the coils of intestine and stomach back into cavity of the abdomen and opening was closed in layers .Post operative period was uneventful Later child was shifted to Neonatal ICU.
CONCLUSION
It is the ultra sound that has played a prominent role in the diagnosis of anomalies in gestation period (antenatal period).Early diagnosis is of paramount importance to obstetrician to plan for further management. Anomalies found after birth also can be to treated either immediately after birth or after certain specified time This case of multiple congenital anomalies with gastroscisis as main anomaly gives knowledge of anomalies not only to Anatomists but to surgeons of all clinical branches and physicians This study of anomaly is of paramount importance to community medicine in bringing down the incidences of morbidity and mortality of anomaly ,through Health workers ,social workers and volunteers ,and NGOs. ANOMALY committee or caell can be set to bring down incidences of Anomalies in coming years . Hence this case has been studied and reported.
ACKNOWLEDGEMENT
Sincere thanks to:-1)Medical Director,.2)Medical Superintendent and 3)Prof and HOD OF Obstretrics and Gynaecology of Teaching Hospital of Raja Rajeswari Medical college Bangalore for permitting me to examine the cases. The immense help received from the scholars whose articles are cited and included in the references of this manuscript .The authors are also grateful to authors/editors/publishers of all these articles ,journals,books,from where the literature for this article has been reviewed and discussed .
Englishhttp://ijcrr.com/abstract.php?article_id=2012http://ijcrr.com/article_html.php?did=20121. Parker.S.E, Mai CT,Canfield MA,Richard.R,Wang Y,Mayer RE, et al :for the National Birth Defects Prevention Network .Updated national birth prevalence estimates for the selected birth in the United States .2004-2006.Bith Defects Res A Clin Mol Teratol .2010 Sept 28 (EPub ahead of print)
2. Yang Ping ,Terri.H Beaty,Muin.J,Khoury,Elsbeth Chee, Walter Stewart and Leon Gordis ‘’Genetic Epidemiologic study of omphalocele and gastrochisis evidence in heterogeneity” American journal Medical genetics 44(2005)668-675;
3. Williams LJ, Kucik JE, Alverson CJ, Olney RS, Correa,A,: Epideomology of Gastroschsis in Metropolitan Atlanta 1968 through 2000 Birth Res A 2005;73:177-83,
4. BirdsTM, Robbins JM, Druschel C, Cleves MA,Yang S, Hobbs C A.and National Birth Defects Prevention study(2009) Demographic and environmental risk factors for gastrochisis and omphalocele in the National Birth Defects Prevention study: .J.Pediatr Surg ;44:1546-1551.
5. Feldkamp ML, Reefhuis J, Kucik J, Krikov S,Wilson A, Moore CA,Carey JC,Botto.LD,and National Birth Defects Prevention study,Case –control study of self reported genitourinary infections and risk of gastrochisis : findings from the National Birth Defects Prevention study-1997 - 2003,BMJ,2008,Jun 21;336(7658);1420-3.
6. YuvaRaj Bhosale,Laxmi Rajgopal, Nandanwar,YS;:Omphalocale,Exstrophy of cloaca ,imperforate anus,and Spinal Defects,(OEIS Complex);A case Report and Review of literature;J.Anat Soc.56.(1) :2007:41—43.
7. MadhavanT and Narayan.J.(1991)- Consanguinity and mental retardation – Journal of Mental Deficiency Research 1991:35:133-139
8. Verma I.C,Prema A,Puri.A.K(1992):-Health effects of Consanguinity in Pondicherry ,Indian pediatrics 1992,,29,685-692
9. Verma I.C,(1978);-High frequency of neural tube defects in North India –Lancet -1978-1- 879-880.
10. Sayee Rajangam ,Rama Devi(2007):- Consanguinity and chromosomal abnormality in mental Retardation or multiple congenital abnormality.-Journal of Anatomical Society of India- Vol56(2),DEC-2007 -30-33p.
11. Sarah Waller MD,Kathleen Paul .MD.,Suzanne Peterson .M.D., and Jane Hitti .M.D.,of the university of Washington,Seattle Society for Maternal – Fetal Medicine (2010,February 7).Link between birth defect gastroscisis and the agricultural chemical atrazine found . Science Daily.Retrieved March 12 2011, from http://www.Science daily.com/releases /2010/02//1000205081805.htm.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareTHE EFFECT OF COMBINED VITAMIN E AND C SUPPLEMENTATION ON THE OXIDATIVE STRESS PARAMETERS IN PATIENTS WITH SCHIZOPHRENIA
English176183Santoshi R. GhodakeEnglish A. N. SuryakarEnglish Abdul Kayyum ShaikhEnglish Prabhakar M. KulhalliEnglish R.K. PadalkarEnglish A. M. RautEnglishRecently, it is proposed that oxidant- antioxidant imbalance may have a role in the pathophysiology of schizophrenia. The present study was performed to assess differences in plasma levels of Malondialdehyde (MDA), as an oxidant product, antioxidant such as superoxide dismutase (SOD), reduced glutathione (GSH), vitamin E (α-tocopherol), vitamin C (ascorbic acid) etc., and total antioxidant capacity(TAC) in the patients with schizophrenia as compared with healthy controls. Our secondary aim was to further evaluate the effect of combined supplementation of antioxidant vitamins E and C along with antipsychotics treatment on these parameters. The study includes prospective, double blind, non-crossover, and 12 weeks intervention of antioxidants. Plasma levels of MDA, erythrocytes SOD, GSH, vitamin E, vitamin C and TAC levels in plasma in patients with schizophrenia before (n=40) and after regular antipsychotic treatment (n=20) and treatment with antipsychotics plus antioxidant E and C supplementation (n=20) were compared with healthy controls (n=40). The levels of MDA were significantly increased in patient before treatment while the levels of SOD, GSH, vitamin E and C and TAC in plasma shown significant reduction. However, the antioxidant supplementation along with antipsychotics reverses the above levels. PANSS (positive and negative syndrome scale) change scores at 12 weeks improved statistically significant with antioxidant E and C supplementation as compared to alone antipsychotics treatment. So oral supplementation of antioxidants in combination reduces oxidative stress and improves clinical symptoms suggest the application of antioxidant in clinical trials is beneficial to prevent or reduces the progression of disease.
Englishoxidative stress, schizophrenia, Treatment, vitamin C, vitamin E.INTRODUCTION
Schizophrenia is a devastating mental disorder, expressed in the form of abnormal mental functions and disturbed behavior. It has a life-time prevalence of approximately 1% of the world?s population. The disorder has a bad outcome, regardless of different treatment (pharmacological, social, cognitive etc.). Oxidative stress is common to several neuropsychiatric disorders. The mechanisms have been most widely studied in schizophrenia, employing various areas of research, including oxidative biomarkers, psychopharmacology and clinical studies with antioxidants (1). The aetiopathogenesis of neuropsychiatric disorders is incompletely understood, which may partly account for the persisting dominance of the syndrome nosology in neuropsychiatry, despite its widely recognized inadequacies. Oxidative stress mechanism have been implicated in the pathogenesis has theoretical appeal, as the brain is considered particularly vulnerable to the damage for several reasons (2). The oxidative vulnerability of the brain, with growing evidence for neuropsychiatry syndromes, suggests that oxidative damage may be a plausible pathogenic candidate (2). In fact, experimental models have demonstrated that oxidative stress induces behavioral and molecular anomalies strikingly similar to those observed in schizophrenia. Thus the extant evidence for the role of oxidative stress in the pathophysiology of neuropsychiatric disorders such as Schizophrenia, Depression, Alzheimer?s disease, Epilepsy etc. offers a hypothesis derived therapeutic approach in the form of antioxidants. Vitamin E and vitamin C are well known antioxidants that are postulated to protect against damage to biological membranes by their ability to scavenge free radicals. Accordingly, several studies have examined the efficacy of vitamin E or vitamin C in the treatment of schizophrenia (1, 3). Treatment of schizophrenia, particularly the most disturbing symptoms is considered to be very difficult. This is because treatment with both typical as well as atypical antipsychotics has serious side effects that lead to a significant loss of quality of life. It is important to develop alternative or adjunctive treatment strategies that may augment the antipsychotic action and reduce the side effects. Clinical trials investigating adjunctive antioxidants in the treatment of schizophrenia and depression have utilized vitamin E and vitamin C, Ginkgo biloba extract and N-acetyl cysteine (NAC) have been considered for some to improve the outcome of these diseases (2). Supplementation has been done, first with vitamin C with haloperidol reported no symptomatic improvement (4). Then vitamins E/C along with omega – 3 fatty acids, shown significant symptomatic improvement, but no significant change in oxidative products has been found (5). Some studies showed reversal of MDA, ascorbic acid and SOD levels along with improved positive and negative symptoms, extrapyramidal side effects and superior BPRS outcome (6,7). As the evidence is increasing for the increased oxidative stress and oxidative cellular injury in various neuropsychiatric disorders, use of more than one antioxidant vitamin may be the preferred supplementation. Thus the use of antioxidants is critical since peroxidative membrane injury may contribute to pathophysiology and must be prevented (8). It is also important to point out that the number of studies has reported variable therapeutic effects of dietary antioxidants, primarily vitamin E on tardive dyskinesis (9). Supplementation with vitamin C, an effective intracellular antioxidant, has not been tried, although its use in preventing intracellular peroxidative injury and restoration of active vitamin E for the prevention of membrane lipid peroxidation has been suggested (8). There is a lack of information on oxidative process in cerebrospinal fluid and brain. Considering the size of the CNS in comparison to the other compartments of the human body, it seems reasonable to assume that changes in the levels of various biomolecules inside the brain can influence the plasma levels of those biomarkers. This was concluded by various studies on CNS (CSF, post mortem or animals) showed at least correlative tendencies with studies assaying biomarkers in erythrocytes, plasma or polymophonuclears (1). So most of the oxidative stress in patients with schizophrenia have made on peripheral tissues. The patients with schizophrenia had increased serum Malondialdehyde levels and decreased plasma ascorbic acid levels, which proves increased oxidative stress. These levels were significantly reversed after treatment with vitamin C along with atypical antipsychotics compared to placebo with atypical antipsychotics. BPRS scores improved significantly with addition of vitamin C as compared to placebo after 8 weeks of treatment (6). The supplementary use of vitamin C in schizophrenia requires caution since a high dietary intake of iron will result in vitamin C having a pro-oxidant rather than an antioxidant action. Vitamin E is a lipid soluble antioxidant with the potential to prevent oxidative damage. However, vitamin E cannot prevent oxidative damage to cytosolic proteins, mitochondria, and nuclei, where most of the ROS are generated. Therefore, it may be important to use vitamin E in combination with vitamin C. Addition of vitamin C probably reduces this risk because of its interactions with vitamin E, i.e. vitamin C reduces vitamin E radicals formed when vitamin E scavenges the oxygen radicals (10). Also, theses vitamins are suitable for human clinical trials because they are readily available, inexpensive and relatively safe (11). We have therefore tested the safety and efficacy of combining vitamin C and E in a sample of patients. So, aim of this study was to explore the beneficial effects of combined supplementation of vitamin E and C on oxidative stress parameters in schizophrenia patients. Also to suggest the new therapeutic approaches using appropriate antioxidants which might partially alleviate or prevent the symptoms of schizophrenia.
MATERIALS AND METHODS
This prospective cohort study, 12 weeks trial was carried out in the department of Biochemistry and outpatient and inpatient department of Psychiatry, PDVVPF?s Medical College, Ahmednagar, India. Study group consist of a control group of healthy subjects and that of schizophrenia selected consecutively. Healthy subjects more than 18 years of age, of both sexes, more than 50 kilograms weight, taking good diet, non-smokers, and non-alcoholic, free of any medication for at least one month prior to study were selected as controls. These were enrolled from the general population and academic community. The research protocol and consent forms were approved by institutional ethics committee. Some the patients had a decision-making capacity and consented for the study signed the consent form, but most of the cases, the consent form was signed by the family members. After taking consent and screening with routine investigations, physical examination. Fasting blood samples were obtained at 0 weeks and at the end of 12th weeks from the start of the treatment. After initial analysis subjects were divided into to two groups by stratified sampling. First group receiving regular antipsychotic treatment (consisting of haloperidol, dose decided by psychiatrist, depends on severity) and second group allow receiving antioxidant vitamins E and C along with antipsychotic treatment. The vitamin supplementation was carried out by supplying both vitamin E (400 IU, dl-α-tocopheryl acetate/day, Evion) and vitamin C (250 mg L-ascorbic acid/day, Celin). The patients were instructed to consume vitamin E and C daily for the duration of 12 weeks (3 months). After 12 weeks of supplementation, the effect of supplements was assessed by measurement of oxidant and antioxidant markers in the patient. The patient and control were excluded for high levels of dietary supplemental, sever under or malnourishment, seizure disorders, head injury, with loss of consciousness, alcohol and substance abuse and dependence, excessive smoking, type II diabetes, lipid disorders, cardiovascular diseases, hypertension or a family history of the same. These factors and obesity, which is also rarely seen in these patients, known to affect free radicals and antioxidant status were also excluded. Diagnosis of schizophrenia was derived by structured clinical interview (SCID-P) according to axis I of DSM-IV (Diagnostic and Statistical manual of Mental Disorders- fourth edition) criteria (12). Patients with WAIS-R full scale IQEnglishhttp://ijcrr.com/abstract.php?article_id=2013http://ijcrr.com/article_html.php?did=20131. Boskvic M, Vovk T, Plesnicar BK, Grabnar I. oxidative stress in schizophrenia. Current Neuropharmacology. 2011; 9: 301-2.
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5. Arvindakshan M, Ghate M, Ranjekar PK, Evans DR, Mahadik SP. Supplementation with a combination of omega-3 fatty acids and antioxidants (vitamins E and C) improves the outcome of schizophrenia. Schizophrenia Research. 2003; 62:195–204.
6. Dakhale GN, Khanzode SD, Khanzode SS, Saoji A. Supplementation of vitamin C with atypical antipsychotics reduces oxidative stress and improves the outcome of schizophrenia. Psychopharmacology. 2005; 182:494–8.
7. Sivrioglu EY, Kirli S, Sipahioglu D, Gursoy B, Sarandol E . The impact of omega-3 fatty acids, vitamins E and C supplementation on treatment outcome and side effects in schizophrenia patients treated with haloperidol: an open- label pilot study. Progress in Neuropsychopharmacology and Biological Psychiatry. 2007; 31:1493–9.
8. Mahadik SP, Evans D, Lal H. Oxidative stress and role of antioxidant and omega-3 essential fatty acid supplementation in schizophrenia. Prog Neuropsychopharmacol Biol Psychiatry. 2001; 25:463-93.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241418EnglishN2012September29HealthcareSERODIAGNOSIS OF DENGUE IN TERTIARY CARE HOSPITAL IN DAVANGERE DISTRICT OF KARNATAKA
English184187G.K.MangalaEnglish G. VishwanathEnglishPurpose: To study the seoprevalence of Dengue infection in Davangere district of Karnataka. Materials and Methods: A total of 1952 blood samples were collected from patients attending tertiary care hospital in south India from January to December 2010.Serum was separated and subjected to Ig M ELISA. Results: Out of 1952 samples tested 583 (29.9%) were positive for IgM ELISA with female preponderance 266 cases (33.29%).Common age group affected was 11-20 yrs followed by 1-10 yrs with 39.45% and 36.36% respectively.Maximum number of cases were seen during the month of july and August. Conclusion: The prevalence of Dengue is 29.9% which is in agreement with other studies . More number of cases were seen in july and august affecting less than 20yrs age group which helps to plan preventive measure strategies.
EnglishDengue, Seroprevalence, ELISA.INTRODUCTION
Dengue fever, dengue heamorrhagic fever (DHF) and Dengue shock syndrome (DSS) represent a spectrum of disease resulting from infection with the dengue virus (member of the genus flavivirus and family flaviviridea)which is primarily transmitted by the mosquito ,Aedes aegypti and Ae albopticus.Dengue fever can be caused by one of four types of dengue virus :DEN-1,DEN-2,DEN-3 and DEN-4.Dengue virus infection is clinically similar to many other acute febrile illnesses ,in which serological testing plays a important role in early diagnosis and management. The spectrum of illness is broad and ranges in severity from mild symptoms to death .The World Health Organisation (WHO) receives reports of about 5,00,000 dengue fever cases each year, but estimates that as many as 50 million people are infected annually with 24,000 deaths(3). India is endemic for dengue infections and various states have reported outbreaks. In India highest number of Dengue cases were reported in 2010, as 25725 cases and 99 deaths till November,2010.In Karnataka highest number were reported in 2010 with 2177 cases and 6 deaths (4) . Laboratory diagnosis of Dengue fever can be done by various methods such as virus isolation and characterisation, detection of genomic sequence by a nucleic acid amplification assay, and detection of dengue virus antigen & specific antibodies (5) .However, most of laboratories in India do not have setup for molecular diagnosis,hence diagnosis is made by demonstration of antigen and antibodies by ELISA based tests. This study was conducted to know the seroprevalence of dengue in tertiary care hospital in central part of Karnataka.
MATERIALS AND METHODS
The study was conducted in department of microbiology, J.J.M. Medical college, Davangere .The study group consist of clinically suspected cases of dengue attending Chigateri General and Bapuji hospital attached to J.J.M.Medical college in the year 2010. A total of 1952 blood samples were collected from clinically suspected cases from January to December 2010.Sera was separated and Ig M ELISA was performed using Dengue IgM Microlisa manufactured by J.Mitra. The test was performed according to manufacturers instruction.
DISCUSSION
The first evidence of occurrence of Dengue fever in the country was reported during 1956 from Vellore district in Tamilnadu . Subsequently , the first DHF outbreak occurred in Calcutta, West Bengal in 1963 with 30% of cases showing haemorrhagic manifestations. All the four serotypes have been isolated in India since 1966 .In the present study seroprevalence of Dengue infection was 29.9% .Similar findings have been reported by various other studies . Ukey PM et al(6) reported prevalence of 31.3%,Lal M(7) et al reported 39.4 %.But Gunasekaran(8) et al reported high prevalence of 43.0% and Garg et al reported low prevalence of 19.7%. In our study high incidence was seen in age group of 11-20 yrs with 230 cases (39.45%) followed by 1-10 yrs with 212 cases (36.36%).This is in concordance with other workers, Gunasekaran(8) et al has reported high incidence of 84.5% in age group less than 14 yrs, Ukey PM et al(6) reported 31.71% in 15-30 yrs age group. The incidence was high in females with 33.29%. Seasonal distribution showed highest incidence in july and August. This is in agreement with other studies like Garg et al(9) , Pandhya G(10) et al and Tripathi P et al(11) .The high incidence in monsoon and postmonsoon is due to the climatic conditions favouring breeding of mosquitos and transmission of disease.
Englishhttp://ijcrr.com/abstract.php?article_id=2014http://ijcrr.com/article_html.php?did=20141. Guzman MG,Kouri G.Dengue diagnosis:advances and challenges.Int J Infect Dis.20004;8:69-80.
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4. Baruah K,Dhariwal AC.Epidemiology of dengue,its prevention and control in India. J Indian Med Assoc.2011;109:82-86.
5. Robert.B.Belshe editor .Flaviviruses in Textbook of human virology,2nd edn,Mosby -Year Book,Inc:1991;23:p 23.
6. Ukey PM,Bondade SA,Paunipagar PV,Powar RM,Akulwar SL.Study of seroprevalence of dengue fever in central India.Indian J Community Med 2010;35:517-19.
7. M Lal,A Aggarwal ,A Oberio.Dengue feveran emerging viral fever in Ludhiana,North India.Indian J Public Health.2007;51:198-99.
8. Gunesekaran P,Kaveri K,Mohana S,Arunagiri K,Babu BS,Priya PP et al. A retrospective analysis Dengue disease status in Chennai (2006-2008) Indian J Med Res . 2011;133:322-25.
9. Garg A,Garg J,Rao YK,Upadhyaya GC,Sakhuja S.Prevalence of dengue among clinically suspected febrile episodes at a teaching hospital in North India. J Infect Dis Immun.2011;3:85-89.
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11. Tripathi P,Kumar R,Tripathi S,Tambe JJ,Venkatesh V.Descriptive epidemiology of Dengue transmission in Uttar Pradesh.Indian Paediatrics.2008;45:315-18.