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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareIN VITRO EVALUATION OF ANTI BACTERIAL ACTIVITY OF HEART WOOD EXTRACT OF ACACIA
CATECHU ON ORAL MICROBES
English0409Geetha R.VEnglish Anitha RoyEnglish Lakshmi TEnglishThe aim of the present study was to evaluate the anti bacterial activity of Acacia catechu on selected oral microbes. Acacia catechu [AC] is a moderate thorny deciduous tree growing in tropical countries. Antibacterial activity of ethanolic and aqueous extract of heart wood of AC is screened against Streptococcus mutans, Streptococcus salivarius, Lactobacillus acidophilus and Enterococcus faecalis using disc diffusion technique. The Minimum inhibitory concentration [MIC] and Minimum bactericidal concentration [MBC] of the extracts were also determined. The results of this study showed that both the extracts at different concentrations exhibited anti bacterial activity against the bacterial species tested. The ethanolic extract showed higher degree of activity than aqueous extract when compared with the standards.
EnglishAcacia catechu ,Anti bacterial ,Disc diffusion , MIC,MBC.INTRODUCTION
Herbal medicines have been used for many years. Their history can be rooted from ancient civilization wherein their role as a primary source of medication is evident. Herbs with medicinal properties are a useful and effective source of treatment for various diseases 1-3. Many of the common drugs used by us today are derived from herbs. Their beneficial effects allied to the current worldwide ?back to Nature? trend and greater attention being paid to these products. The development of drug resistance in human pathogens against commonly used antibiotics has necessitated a search for new antimicrobial substance from other sources including plants.4 Currently many studies are being conducted to know these herbs in depth. Screening of medicinal plants for antimicrobial activities are important for finding potential new compounds for therapeutic use. There have been numerous reports of the use of traditional plants and natural products for the treatment of oral diseases. Many plant-derived medicines used in traditional medicinal systems have been recorded in pharmacopeias as agents used to treat infections and a number of these have been recently investigated for their efficacy against oral microbial pathogens. The present study was to evaluate the antibacterial activity of heart wood extract [ethonolic and aqueous] of Acacia catechu willd on selected oral microorganisms.
Acacia catechu (AC) (Family: Fabaceae and subfamily: Mimosoideae)
known as Black cutch..AC is medium sized thorny deciduous tree mainly found in India and also found in deciduous forests around the world. It grows up to 13 meters in height It is said that the name ?catechu' was given to it because its bristles resemble the claws of animals of the cat family or may be because its heart wood contains gummy extract called kath or cutch. The foliage is softly textured, light green and oval-shaped. The branches are thin and spike like due to tiny thorns grown around the exterior. The sap wood of AC is large and yellowish white and heart wood is small and red in colour. The chief constituents of the heartwood are catechin and catechutannic acid 5 . The wood contains epicatechin 5 , Atzelchin, catechin tetramer, dicatechin, gallochin, gossypetin, phlobatannin, kaempferol, quercitrin, quercitin 6,7,8. Catechin is biologically highly active. It is used as a haemostatic. Taxifolin an other important constituent has antibacterial8 , anti-fungal12, antiviral, anti-inflammatory, and antioxidant activity6 . The extract of Acacia catechu extract have been reported to have various pharmacological effects like immuno modulatory13, anti pyretic 7 , hypoglycaemic7 , anti diarrhoeal7, hepatoprotective activity7,11. Cutch is astringent, cooling, and digestive. It is useful in cold and cough9,10 ulcers, boils and eruptions of the skin, bleeding piles, uterine haemorrhages, atonic dyspepsia, chronic bronchitis etc. In stomatitis, halitosis, dental caries and cavities16, AC is used with great benefit. An antibacterial mouthwash made from the extract treats gingivitis and mouth sore The leaves, bark, heartwood has many nutritional and medicinal uses.
MATERIALS AND METHODS
Plant material
The ethanolic and aqueous extract of heartwood of Acacia catechu willd was obtained from Green Chem Herbal Extract and Formulations. Bangalore.
Test microorganisms
Bacterial strains used were Streptococcus mutans [Gram positive cocci], Streptococcus salivarius [Gram positive cocci], Enterococcus faecalis [Gram positive cocci], Lactobacillus acidophilus [Gram positive bacilli]., The organisms were obtained from department of Microbiology, Saveetha Dental College and maintained in nutrient agar slope at 4C
METHODOLOGY
The extracts were prepared in the following concentrations in sterile water. 1mg/ml, 2mg/ml, 3mg/ml and 4mg/ml. 50µl of extract of different concentrations were loaded on sterile filter paper discs measuring 6mm in diameter, so that the concentration of the extract on each disc was 50µg, 100µg, 150µg and 200 µg respectively. The discs were dried and kept aseptically. Screening of antibacterial activity [Disc diffusion technique] Broth culture of the bacterial strains compared to Mac Farland‘s standard 17-20 0.5 were prepared. Lawn culture of the test organisms were made on the Muller Hinton agar [MHA-Hi media M1084] plates using sterile cotton swab and the plates were dried for 15 minutes.Filter paper discs loaded with different concentrations of the extract were placed on the respective plates. The plates were incubated at 37°C overnight and the zone of inhibition of growth was measured in millimeters. Standard antibiotic discs of amoxicillin 6 International Journal of Current Research and Review www.ijcrr.com Vol. 03 issue 06 June 2011 (30mcg/disc) and PenicillinG (30mcg/disc) were used as positive control. All the tests were done in triplicate to minimize the test error.
Determination of minimum inhibitory concentration
Macro broth dilution or tube dilution method was done to determine the Minimum inhibitory concentration (MIC) of the extracts17-20. A series of two fold dilution of each extract ranging from 4mg/ml to 0.125mg/ml was made in Muller Hinton broth as specified by National Committee for Clinical Laboratory Standards (NCCLS, 1998). 100μl of standard inoculum of the bacterial strains matched to 0.5 Mc Farland‘s standard were seeded into each dilution. Two control tubes were maintained for each test batch. These included antibiotic control (tube containing extract and growth media without inoculum) and organism control (tube containing the growth medium and the inoculum) .The tubes were incubated at 37°C for 24 hours and checked for turbidity. MIC was determined as the highest dilution (that is, lowest concentration) of the extract that showed no visible growth..
Minimum Bactericidal Concentration (MBC)
The MBCs were determined by selecting tubes that showed no growth during MIC determination; a loop full from each tube was sub cultured onto Muller Hinton agar plates and incubated for further 24 hours at 37oC. The least concentration, at which no growth was observed, was noted as the MBC.
RESULT AND DISCUSSION
The antibacterial activity of the extracts (Ethanolic and Aqueous) at different concentrations was screened by disc diffusion technique and the zone of inhibition was measured in mm diameter. The results are given in the table 1. The minimum inhibitory concentration [MIC] and minimum bactericidal concentration [MBC] were also determined for the extracts and the results are given in table 2. The ethanolic extract was more effective against Streptococcus mutans with a zone of inhibition of 24 mm diameter (at conc 200 µg.) and was least effective against Lactobacillus acidophilus with zone of inhibition of 16 mm (at conc. 200 Pg.) and among the other bacterial species studied Streptococcus salivarius showed a zone of inhibition of 19mm diameter (at conc. 200 Pg.) and Enterococcus faecalis showed inhibition zone of 22 mm diameter (at conc. 200 Pg.). The MIC and MBC values of ethanolic extract was found to be low compared to aqueous extract. The ethanolic extract was found to have Low MIC and MBC values of 0.5mg/ml and 0.5mg/ml and 0.5mg/ml and 1mg/ml for Streptococcus mutans and Enterococcus faecalis respectively. With Lactobacillus acidophilus ethanolic extract showed a higher MIC and MBC value of 2mg/ml and 2mg/ml and for Streptococcus salivarius it was 1mg/ml and1mg/ml. The lower MIC and MBC value is an indication of high effectiveness of the extract whereas higher MIC and MBC indicates the less effectiveness of the extract.
Oral diseases are major health problems with dental caries and periodontal diseases among the most important preventable global infectious diseases. Oral health influences the general quality of life and poor oral health is linked to chronic conditions and systemic diseases. The association between oral diseases and the oral microbiota is well established. Dental caries is a microbial disease that result in the destruction of mineralized tissue of the teeth. Streptococcus mutans is the potent initiator and leading cause of dental caries world wide 21. It is considered to be the most cariogenic of all of the oral Streptococci. Another organism that is important in the development of caries is Lactobacillus acidophilus. This bacteria is not important in the initiation of caries but in the continuation.21 The other problem faced in the field of dentistry is failure of endodontic treatment. Enterococcus faecalis is the commonly isolated bacteria from the infected root canal and being considered a possible cause of failure in root canal treatment 22. It is one of the most resistant species that can best adapt to and tolerate the ecologically demanding conditions in the filled root canal. The present study was to evaluate the antibacterial activity of Acacia catechu against caries causing organisms and organism associated with endodontic infections. The bacterial strains used in this study were strongly associated with caries, Streptococcus.mutans being the most cariogenic bacteria. Streptococcus salivarius and Lactobacillus acidophilus were also found to cause caries. Enterococcus faecalis is responsible for most of the endodontic infections. The results obtained from our study shows that ethnolic extract has got a very good antibacterial activity against the selected oral pathogens.
CONCLUSION
The present results therefore offer a scientific basis for traditional use heartwood extract of Acacia catechu on oral pathogens. The anti-bacterial activities could be enhanced if active components are purified and adequate dosage determined for proper administration. The dentist needs to be informed regarding the herbal and over-the-counter products that may impact the delivery of safe and effective dental treatment. The use of herbs in dentistry should be based on evidence of effectiveness and safety.
ACKNOWLEDGEMENTS
The authors are thankful to Mr.Rajendran.R, Green Chem PVT LTD. Bangalore for providing us the Ethanolic extract of heartwood of Acacia Catechu Willd as a gift sample for our research work. We wish to thank H.O.D of Microbiology, Saveetha Dental College. Dr. Selvaraj, Assistant Professor microbiology ,Saveetha Dental College.
Englishhttp://ijcrr.com/abstract.php?article_id=2155http://ijcrr.com/article_html.php?did=21551. Sheetal verma and S.P.Singh., Current and future status of herbal medicines. Veterinary world, Vol.1(11): 347 – 350.
2. R.Guo.,P.H.Canter.,E Ernst., A systematic review of randomised clinical trials of individualised herbal medicine in any indication.Post graduate medical journal. 2007 ;83 633-637.
3. Enzo A. Palombo.,Traditional Medicinal Plant Extracts and Natural Products with Activity against Oral Bacteria: Potential Application in the Prevention and Treatment of Oral Diseases Review article. Evidence – Based complementary and Alternative Medicine Vol 2011,Article ID680354,15 pages
4. Si-Yuan Pan, Si-Bao Chen, HongGuang Dong, Zhi-Ling Yu, Ji-Cui Dong, Zhi-Xian Long,Wang-Fun Fong, Yi-Fan Han, and Kam-Ming Ko.,New perspectives on Chinese herbal medicine research and development. Evidence Based Complementary and alternative medicine volume 2011.,11 pages.
5 Rao PR, Seshadri TR, L-Epi-catechin from Acacia catechu, Journal Scientist Indian Research, 7B, 1948, 59
6 V. Gayathri devi, Anitha John, R.Sreekala devi, V. A. Prabhakaran .Pharmacognostical studies on acacia catechu willd and identification of journal of pharmacy and pharmaceutical sciences. vol 3, suppl 2, 2011
7 Ray D, Sharatchandra KH, Thokchom IS, Antipyretic, antidiarrhoeal, hypoglycaemic and Hepatoprotective activities of ethyl acetate extract of Acacia catechu Willd. In albino rats, Indian Journal of Pharmacology, 38(6), 2006, 408-413.
8 Bhawna sunil and Bharti P Dave. In vitro anti microbial activity of Acacia catechu and its phytochemical Analysis. Indian Journal of Microbiology, Vol 50 Number 4, 369-374
9 Anonymous, The Wealth of India, Raw Material, Vol 1, CSIR, New Delhi, 2004, 11.
10 Wallis TE, Textbook of Pharmacognosy, 5th Edition, CBS Publishers and Distributors, New Delhi, 2005, 461-463.
11 Jayasekhar P, Mohanan PV, Hepatoprotective activity of ethyl acetate extract of Acacia catechu, Indian Journal of Pharmacology, 29(6), 1997, 426-428
12 Nagaraja T.G *, S.V. Sarang and D. C. Jambhale Evaluation of anti-mycotic activity of Acacia catechu Willd.(Mimosaceae) Anti-mycotic activity of Acacia catechu Journal of Biopesticides,1(2):197 - 198 (2008)
13 Syed Ismail and Mohammed Asad., Immunomodulatory activity of Acacia catechu. Indian journal of Pharmacology 2009:53 (1), 25 -33
14 Ammara Hassan, Salma Rahman, Farah Deeba and Shahid Mahmud. Antimicrobial activity of some plant extracts having hepatoprotective effects .Journal of Medicinal Plants Research Vol. 3(1), pp. 020-023, January, 2009
15 Pramod, Shahid H Ansari and Javed Ali., Herbal Remedies for the Treatmebt of Periodontal Disease – A patent review. Recent patents on Durg delivery and formulations 2009, 3, 221 - 228
16 J. Smullen, G.A. Koutsou, H.A. Foster, A. Zumbé, D.M. Store The Antibacterial Activity of Plant Extracts Containing Polyphenols against Streptococcus mutans. Caries research Vol. 41, No. 5, 2007
17 Collins,CH and Lyne, P.M 1976.Microbiological methods, London, Butterworths and co.288p
18 Betty A.Forbes., Daniel F.Sahm., Alice S.Weissfeld. Bailey and Scott‘s Diagnostic Microbiology 11th edition Mosby page 229 - 257
19 Connie R.Mahon., George Manuselis., Saunder‘s Diagnostic Microbiology 2nd Edition
20 J.G.Colle., A.G.Faster., B.P.Marmion., A.Simmons. Practical Microbiology [Mackie and Mc Cartney] 14th edition page 851 - 852
21 L.P. Samaranayake.,Brain M.Jones., Essential Microbiology for Dentistry. Second edition page 217 - 223.
22. Isabelle portenier., Tuomos M.T. Waltmo., Markus haopaslo., Enterococcus faecalis- the root canal survivor and ?star' in post treatment disease. Endodontic Topics 2003,vol- 6,135- 159
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30General SciencesIMPROVED GROUP ACCEPTANCE SAMPLING PLAN FOR MARSHAL-OLKIN EXTENDED LOMAX DISTRIBUTION PERCENTILES
English1025Muhammad AslamEnglish Muhammad ShoiabEnglish Y.L. LioEnglish Chi-Hyuck JunEnglishIn this paper, a group acceptance sampling plan, based on the number of failures occurred from all groups, is proposed for Marshal-Olkin extended Lomax distribution percentiles under the truncated life test. The minimum number of groups needed for a given group size and acceptance number are determined by using the two-point approach, that meets the consumer‘s and producer's risks at the specified percentile quality levels, respectively. The advantages of the designed plan are discussed by comparing with original group acceptance plan proposed by Aslam and Jun (2009). The tables are constructed and results are explained with examples.
EnglishGroup acceptance sampling plan; Marshal- Olkin extended Lomax distribution, consumer‘s risk, producer‘s risk, Truncated life test.1. INTRODUCTION
Acceptance sampling scheme is concerned with inspecting the product and making the decision by using the sample information. Accepting sampling is one of the major techniques in SQC to assure the quality of the product. It is widely used in the field of industry and business to manage the product reliability and to make the decision of accepting or rejecting the product by the consumer. Today, products are designed to be with high reliability, and most of them suffer destructive life tests to collect life information. In contrast with 100% inspection, acceptance sampling plans have the advantage to reduce the cost and meanwhile to ensure that the product quality meets the standard of the consumer. The essential technique of acceptance sampling is to establish the minimum number of sample items for the test. Due to the fact that it could be difficult to observe the complete lifetime of a high reliability item, the truncated life test would be combined with a sampling plan to save time, cost, efforts, manpower, electricity of experiment. Time truncated life test for a various distributions were discussed by Epstein (1954), Goode and Kao (1961), Kantam and Rosaiah (1998),Kantam et al. (2001), Balkizi (2003), Rosaiah et al. (2006), Rosaiah and Kantam (2005), Tsai and Wu (2006), Rosaiah et al. (2007), Aslam and Kantam (2008), Balakrishnan et al. (2007), Aslam and Jun (2010) and Radhakrishnan and Vinotha (2011).
In many situations, putting a number of items in a tester for life test could save time and cost more. In this life test, the tester is called the group and the number of items in each tester is called the group size. An acceptance sampling plan with these groups of items is called the group acceptance sampling plan. The testers had been used in the sudden death testing by Pascual and Meeker (1998) discussed how sudden death experiment could significantly reduce testing time and still yield the estimates of the Weibull percentiles that were as precise as when all sample lifetimes used. Jun et al. (2006) also studied variable sampling plans for Weibull distribution under the sudden death testing. More recently, Aslam and Jun (2009) proposed the group acceptance sampling plan for life test under the inverse Rayleigh or log-logistic distribution. Rao (2010) developed the Marshal-Olkin extended Lomax distribution for a group acceptance sampling plan based on truncated life test. In these papers, using the single point approach to satisfy the consumer‘s risk, the producer‘s risk was not considered. Traditionally, acceptance plans are designed based on the mean lifetime of items for assuring the quality level of the product. However, the sampling plans based on the population mean may not catch the specific percentile of lifetimes asked from the designed considerations of engineering. According to Lio et al. (2010) when the quality of interest is a low percentile, the sampling plans based on the population mean could pass the lot but the required low percentile is below the prespecified standard asked from the customer. Furthermore, a small decrease in the mean with a simultaneous small increase in variance can result in a significant downward shift in small percentiles of interest. This means that a lot of products could be accepted due to a small decrease in the mean lifetime after inspection. But the low percentile of lifetime for products in the accepted lot may not meet the consumer‘s expectation. Lio et al. (2010a, 2010b) introduced acceptance sampling plans based on truncated life test for Birnbaum- Saunders percentiles and Burr Type XII percentiles, respectively Two risks in acceptance sampling plan play a vital role for assuring the quality of the product. Consumer‘s and producer‘s risks are fairly well defined in terms of good product rejected and bad product accepted. In this paper, the two-point approach, which had been used for designing variable acceptance sampling plans by Fertig and Mann (1998) and by Jun et al. (2006), is used to design the proposed group acceptance sampling plan toprotect the consumer‘s and produce‘s risks. The main purpose of this paper is to develop the group acceptance sampling plan under the truncated life test for the percentile of product lifetimes which follow the Marshal-Olkin extended Lomax distribution by using the two-point approach.
2. Introduction of distribution
Assume that the product lifetimes follow the Marshall-Olkin extended Lomax distribution, whose probability density function (pdf) and cumulative distribution function (cdf) are, respectively, given by:
Ghitany et. al. (2007) investigated properties of the Marshal-Olkin extended Lomax distribution and fitted the distribution to a randomly censored lifetimes. Since then, the Marshal-Olkin extended Lomax distribution has been applied to model the product lifetime and the acceptance sampling plans; for examples, Srinivasa Rao et al. (2008) and Srinivasa Rao et. al. (2009) studied single acceptance sampling plan for the mean of the Marshal-Olkin extended Lomax distribution.
3. Group Sampling Plan Based on Total Number of Failures
The consumer‘s risk, , is used as the significance level for this hypothesis testing and 1 is called the consumer‘s confidence level. The proposed group acceptance sampling plan under the total number of failures is stated below:
1. Draw the random sample of size n from a lot, allocate r items to each of g groups (or testers) so that n rg and put them on test until the pre-determined time 0 t .
2. When the test is terminated at the predetermined time 0 t , the lot is accepted if the number of failures from g groups is smaller than or equal to c ; otherwise, the lot is rejected.
3. The lot is rejected and the life test is truncated before 0 t as soon as the total number of failures from g groups is larger than c before 0 t . Since the lot of products is accepted only if the number of failures from all groups is smaller than or equal to the specified acceptance number c, the lot acceptance probability can be calculated as,
It can be seen that the solutions for g and c are independent of the scale parameter . It should be noticed that the failure probability, 1 p , at the quality level of the consumer‘s risk is called the lot tolerance reliability level (LTRL) and the failure probability, 2 p , at the quality level of the producer‘s risk is called the acceptable reliability level (ARL).
4. Description of Tables and Examples
Given group size, r 5, or 10 , and the termination time multiplier q 0.5 or 1.0 , the minium number of groups g and the acceptance numbers c required for the proposed group acceptance sampling plans to ensure the 100-qth percentile life, where q 0.1, 0.5 , have been obtained under various consumer‘s risk values ( 0.25,0.10,0.05,0.01) at the true 100-qth percentile equal to the specified life and the producer‘s risk value 0.05 at the true 100-qth percentile ratios 2 d = (4,6,8,10,12). The results for population parameters (v 3, 3), (v 2, 3) and (v 3, 4) are given in Tables 1-6. Tables 1-3 provide the group acceptance sampling plans for 10 percentile life and Tables 4-6 show the group acceptance sampling plans for median life. Let the product lifetimes have the MarshalOlkin extended Lomax distribution with parameters 3 and 3 . To protect the consumer‘s risk of 0.25 at the prespecified 10 percentile life 0 q t and the producer‘s risk of 0.05 at the true 10 percentile life equal to 4 times of 0 q t , Table 1 shows that the group sampling plan should use 21 testers with 5 items in each tester and the acceptance number c 3 , if the life test would be terminated at 0 0.5 q t . To protect the consumer‘s risk of 0.25 at the pre-specified median life 0 q t and the producer‘s risk of 0.05 at the true median life equal to 4 times of 0 q t , Table 4 shows that the group sampling plan must use 5 testers with 5 items in each tester and the acceptance number c 4 , if the life test would be terminated at 0 0.5 q t . Similarly, given the same protections for consumer and producer as mentioned above, assuming the product lifetimes have the Marshal-Olkin extended Lomax distribution with parameters 2 and 3 , Table 2 indicates that the group sampling plan needs 20 testers with 5 items in each tester and the acceptance number c 3 , if the life test would be terminated at 0.5 multipliers of specified 10 percentile life 0 q t . In viewing Tables 1-6, it could be noticed that when the ratio 0 / q q t t increases, the number of groups and the acceptance numbers decrease at the same time.
Example 1.
Suppose that the lifetimes of products follow the Marshal-Olkin extended Lomax distribution with shape parameters v 2, 3 . It is desired to design a group acceptance sampling plan to test that the median life is greater than 2000 hours and manufacturer wants to run an experiment for 1000 hours using testers equipped with 5 products each. Let us assume that the consumer‘s risk is 0.25 and the producer‘s risk is 5% when the true percentile is 12,000 hours. Since v 2, 3, 0.25,r 5, q 0.5,and / 6 0 q q t t for this example, the minimum number of groups and acceptance can be found as g 3,c 2 from Table 5. This indicates that 15 products are needed for life testing and that 5 products are allocated to each of 3 testers. We will accept the lot if no more than 2 failures observed before 1000 hours in 3 groups. Example 2. In this example, the data set of successive failure intervals for the air conditioning system of jet plane 8044 reported by Proschan [21] is used. The data set contains 12 observations, 487, 18, 100, 7, 98, 5, 85, 91, 43, 230, 3, 130 (in hours). Fitting this data with the Marshal-Olkin extended Lomax distribution, Ramesh C. Gupta (2009) provided the shape parameter 3.5750 and the index parameter 1.4476 . The number of groups and acceptance numbers for this case of Marshal-Olikin extended Lomax distribution are given in Table 7 for 0.25, 0.05 and 10-th percentile.
5. Comparison
Figure 1 and Figure 2 present the comparison in terms of the number of groups needed between the proposed plan and the original group acceptance sampling plans proposed by Aslam and Jun (2009). Both figures indicate that the proposed sampling plan is better than the original sampling plan proposed by Aslam and Jun (2009), since the number of groups needed for the proposed sampling plan is smaller than the number of groups needed for the sampling plan proposed by Aslam and Jun (2009), given a percentile ratio 0 / q q t t . Figure 3 and Figure 4 present the comparison again in terms of the number of groups needed between Marshal-Olkin extended Lomax distribution and generalized log-logistic distribution. It can be noticed that Figure 3 and Figure 4 show the proposed distribution is better than the generalized log-logistic distribution because the number of groups needed for the case of Marshal-Olkin extended Lomax distribution is smaller than the number of groups needed for the case of generalized log-logistic distribution, given a percentile ratio 0 / q q t t .
5. CONCLUSION
In this paper, we proposed the Group acceptance sampling plan based on the number of failures from all groups for Marshal-Olkin extended Lomax distribution percentile lifetime. The two point approach was adopted to determine the required minimum number of groups and acceptance numbers. This paper only deals with the Marshal-Olkin extended Lomax distribution to check the multiple numbers of items simultaneously by saving cost and time. This paper indicate that the proposed plan is better than the original plan (stated by Aslam and Jun (2009)) because the number of groups is smaller as compared to the original plan proposed by Aslam and Jun (2009). This paper also indicates that the Marshal-Olkin extended Lomax distribution is better than the generalized log-logistic distribution because the number of groups is smaller for the case of Marshal-Olkin extended Lomax distribution. The present plan can be used to inspect the many electronic things including for example air conditioning system of jet plane.
Englishhttp://ijcrr.com/abstract.php?article_id=2156http://ijcrr.com/article_html.php?did=21561. Aslam, M., and Jun, C.-H. (2009). A group acceptance sampling plans for truncated life tests based on the inverse Rayleigh and log-logistic distribution. Pakistan Journal of Statistics, 25, 1-13.
2. Aslam, M., and Jun, C.-H. (2010). A double acceptance sampling plan for generalized log-logistic distributions with known shape parameters. J. App. Statist., 37(3), 405-414.
3. Aslam, M., and Kantam, R.R.L. (2008). Economic reliability acceptance sampling based on truncated life tests in the BirnbaumSaunders distribution. Pakistan Journal of Statistics, 24, 269-276.
4. Baklizi, A. (2003). Acceptance sampling based on truncated life tests in the Pareto distribution of the second kind. Advances Appl. Statist., 3(1), 33- 48.
5. Balakrishnan, N., Leiva, V. and Lopez, J. (2007). Acceptance sampling plans from truncated life tests based on the generalized Birnbaum-Saunders distribution. Comm. Statist. - Simu. And Compu., 36, 643-656.
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7. Fertig, F.W., and Mann, N.-R (1980). Life test sampling plans for two parameter Weibull distributions. Technometrics, 22(2), 160-167.
8. Ghitany, M. E., Al-Awadhi, F. A., Alkhalfan, L. A. (2007). Marshal-Olkin extended Lomax distribution and its application to censored data. Communication in Statistics Theory and Methods, 36, 1855-1866.
9. Goode, H.P. and Kao, J.H.K. (1961). Sampling plans based on the Weibull distribution. In Proceeding of the Seventh National Symposium on Reliability and Quality Control, (24- 40). Philadelphia.
10. Jun, C.-H., Balamurali, S. and Lee, S.- H. (2006). Variables sampling plans for Weibull distribution lifetimes under sudden death testing. IEEE Transactions on Reliability 55, 53-58.
11. Kantam, R.R.L. and Rosaiah, K. (1998). Half logistic distribution in acceptance sampling based on life tests, IAPQR Transactions, 23(2), 117- 125.
12. Kantam, R.R.L., Rosaiah, K. and Rao, G.S. (2001). Acceptance sampling based on life tests: Log-logistic models. J. App. Statist., 28(1), 121- 128.
13. Lio, Y.L., Tsai, Tzong-Ru and Wu, Shuo-Jye. (2010a). Acceptance sampling plans from truncated life tests based on the Birnbaum-saunders distribution for Percentiles. . Comm. Statist. - Simu. And Compu., 39(1), 119-136
14. Lio, Y.L., Tsai, Tzong-Ru and Wu, Shuo-Jye. (2010b). Acceptance sampling plans from truncated life tests based on Burr type XII percentiles. Journal of Chinese institute of Industrial Engineers., 27(4), 270-280.
15. Pascual, F.G. and Meeker, W.Q. (1998). The modified sudden death test: planning life tests with a limited number of test positions. Journal of Testing and Evaluation 26, 434-443.
16. Radhakrishnan., R. and Vinotha, P. (2011). Construction of repetitive group sampling plan indexed through producer‘s nano quality level and consumer‘s nano quality level, International Journal of Current Research and Review, 2 (2), 62-66
17. Rosaiah, K. and Kantam, R.R.L. (2005). Acceptance sampling based on the inverse Rayleigh distribution. Eco. Quality Control, 20(2), 277-286.
18. Rosaiah, K., Kantam, R.R.L. and Santosh Kumar, Ch. (2006). Reliability of test plans for exponentiated loglogistic distribution, Eco.Quality Control, 21(2), 165-175.
19. Rosaiah, K., Kantam, R.R.L. and Santosh Kumar, Ch. (2007). Exponentiated log-logistic distributionAn economic reliability test plan. Pak. J. Statist., 23(2), 147-146.
20. Srinivasa Rao, G., Ghitany, M.E., Kantam, R.R.L. (2008). Acceptance sampling plans for Marshal-Olkin extended Lomax distribution. International Journal of Applied Mathematics,
21 (2), 315-325. 21. Srinivasa Rao, G., Ghitany, M.E., Kantam, R.R.L. (2009). Acceptance sampling plans for Marshal-Olkin extended Lomax distribution. International Journal of Applied Mathematics, 22 (1), 139-148.
22. Tsai, Tzong-Ru and Wu, Shuo-Jye (2006). Acceptance sampling based on truncated life tests for generalized Rayleigh distribution. J. App. Statist., 33(6), 595-600.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30General SciencesENDOPHYTIC AND SYMBIOTIC MYCOTROPHY IN EQUISETUM ARVENSE L.: A MEDICINAL SPOREDISPERSING VASCULAR SPOROPHYTE
English3341Vipin ParkashEnglish Priya DhunganaEnglishArbuscular mycorrhizae (AM) are one of the most widespread and common type of symbiotic associations. Equisetum arvense L. is an important medicinal species of seedless vascular plants used in Khasi Hills under Nongkhyllem Reserve Forest, Nongpoh, Meghalaya, India. The mycorrhizal status of this species is meager and scarce in literature. In this study, typical AM structures were observed in sporophytes. The percentage of root length colonized by AM fungi was ranged from 80-90%. The morphological AM colonization pattern was typical Paris-type in rhizome part whereas in young roots (rhizoids), an Intermediate type of infection (both Paris and Arum types) was present. True vesicles were not seen but typical arbusculate and vesciculate structures, microsclerotia and cynophycean filamentous/trichomes structures were seen associated in the rhizoids of sporophyte. Dark septate endophyte (DSE) infection was only seen in rhizome part of sporophyte rather than rhizoids in which fine endophytes as well as endomycorrhizal infection were present. This study depicts that Equisetum rhizoids host abundant and diverse endorhizal fungal associates and also the evolution of Equisetum arvense sporophyte from hydric to mesic habitation due to its typical Intermediate type of endophytic and symbiotic association.
EnglishINTRODUCTION
The vascular plants are divided artificially into two major groups, the seedless (or sporedispersing) vascular plants and the seed plants. There are four major Phyla of spore-dispersing vascular plants: Psilotales, Lycophyta, Equisetales and Pterophyta. The first three Phyla, often referred to as the "fern allies" are not prevalent organisms in our ecosystems today although they are well represented in the fossil record. All of the vascular plants have a dominant sporophyte generation, and a reduced, often, dependent gametophyte stage. Equisetum is the sole surviving genus of ClassEquisetopsida (Sphenopsida). Equisetum arvense Linn. (Family–Equisetaceae, OrderEquisetales) is commonly known as Horsetail. In Khasi hills, Meghalaya, it is known as ?Teengthei’ in local dialect. The name Equisetum is derived from the Latin roots equus, meaning "horse," and seta, meaning "bristle". Horsetail is a non-flowering spore dispersing vascular plant found throughout Europe, Asia, the Middle East, and North America. The plant is a perennial with hollow stems and shoots that look like Asparagus species. As the plant dries, silica crystals that form in the stems and branches look like feathery tails and give the plant a scratching effect that accounts for its name ?scouring rush‘ and its historic use in polishing of metals, particularly pewter. It was used traditionally by Khasi people to stop bleeding, heal ulcers and wounds and treat injury problems. Horsetail contains silicon, which plays a role in strengthening bone. For that reason, it is sometimes suggested as a treatment for osteoporosis. It is also used as a diuretic, and as an ingredient in some cosmetics. Horsetail is used as a diuretic; has haemostatic properties; also considered useful in dropsy, gravel and kidney affection; the ash of the plant is useful in acidity of the stomach and in dyspepsia (Sandhu et al., 2010). It is also a ?metallophyte‘ and indicates the presence of gold and silica mineral in the soil (Nemec et al., 1936). Seed less vascular plants including Lycophytes and Monilophytes (Equisetales) are of ancient origin and occupy a very important position in the origin and evolution of vascular plants (Remy et al., 1994). Mycotrophic and symbiotic status of Lycophytes and Monilophytes is meager and not much known (Dhillion, 1993; Zhao, 2000; Zhang et al., 2004). There are some research papers in which such association was found lacking or absent due to evolution of sporophyte (Treu et al., 1996; Schmidt and Oberwinkler, 1993; Berch and Kendrick, 1982; Malloch et al., 1980; Read et al., 2000). A review of the literature also reveals that work on Equisetum roots is sparse (Bierhorst,1958; Hauke, 1978). But Nasim et al.,1987; who concluded that Equisetum hosted abundant AM fungi (but claimed to have found AM structures in above-ground and in dead plant parts only), some researchers like Dhillion (1993); Koske et al. (1985); reported moderate to little endorhizal colonization in Horsetail. Although, Muthukumar and Udaiyan (2000); have studied the occurrence of AM interactions in some pteridophytes on Western Ghats of India, but very little or almost no work was found in North-East Indian Pteridophytes particularly in Khasi Hills under Nongkhyllem Reserve Forest, Nongpoh, Meghalaya, India. So the aim of the present study was to analyze the occurrence and abundance of mycorrhizal and other endophyts in sporophytes of E. arvense growing naturally in Khasi Hills under Nongkhyllem Reserve Forest, Meghalaya, India for symbiotic mycotrophy
MATERIALS AND METHODS
Study Area
This study was carried out in Nongkhyllem Reserve Forest, located near Nongpoh, Meghalaya, India (250 5′-260 10′ N latitude, 890 47′-920 47′ E longitude). Nongpoh is the district- headquarter of Ri-Bhoi district of Meghalaya and it is situated on GuwahatiShillong National Highway (NH-40). Nongkhyllem Reserve Forest, Nongpoh is having an area of 96 km2 excluding the 29 km2 area of Nongkhyllem Wildlife Sanctuary. Due to the diverse climatic and topographic conditions, Nongkhyllem Reserve Forest supports a vast floral diversity, including a large variety bryophytes, pteridophytes, epiphytes, succulent plants shrubs and trees
Soil sample collection
Rhizospheric soil samples from the roots/rhizomes were collected from road-side stream and moist and damp bench during August-September, 2010 by digging out a small amount of soil close to plant roots up to the depth of 15-30 cm and these samples were kept in sterilized polythene bags at 5-10C for further processing in the laboratory for mycorrhizal quantification and root colonization.
Isolation of spores
Isolation of VAM spores was done by wet sieving and decanting (Gerdemann and Nicolson, 1963) technique. Approximately 100gm of soil were suspended in 1 liter or more of water. Heavier particles were allowed to settle for a few seconds and the liquid was decanted through sieves of different sizes in the order 150µm, 120µm, 90µm, 63µm, 45µm which removes large particles of organic matter, but course enough to allow the desired spores to pass through. The seivings retained on different sieves were collected on different Petri dishes then the trapped spores were transferred to Whatman filter paper no. 1 by repeating washing with water. The spores were picked by hydrodermic needles under stereo- binocular microscope.
Mycorrhizal quantification
For quantitative estimation of VAM spores, Gaur and Adholeya modified method (1994) was used. The filter paper was divided into many small sectors by marking with a ball pen. The total number of spores was counted by adding the number of spores present in each sector under stereo-binocular microscope.
Identification of VAM fungi
For identification of VAM spores the following criteria were used like conventional morphological character i.e. color, size, shape wall structure, surface, ornamentation of spores, nature and size of subtending hyphae, bulbous suspensor, the number and arrangement of the spores in the sporocarp. These VAM spores were identified by using the keys of Schenck and Perez (1990); Morton and Benny (1990), and Mukerji (1996). Clearing and staining of root segments Rhizospheric soil samples were stained according to Phillips and Hayman (1970) method. Roots were first cleared with 10% (w/v) KOH at 980C for one hour. After clearing, roots were bleached with a 5% H2O2 (v/v) solution for 10 minutes. Equisetum roots were darker, so they were bleached in this solution for 40 minutes. All samples were acidified with 1% (v/v) HCl for 5 minutes and stained with 0.05% (w/v)Trypan Blue in acidic glycerol by heating them at 980C for 15 minutes. The stained roots were stored in acidic glycerol for further study.
Analyses of the root samples
Ten stained root pieces approximately 1 cm long were mounted on a slide in Glycerol and were examined with a light trinocular microscope (Labovision, BIOXL). A total of three replicates were made. Typical structures that indicated the presence of mycorrhizas or other root associated fungi, cynophycean filamentous like structures were documented and microphotographed with light trinocular microscope attached digital camera (Canon A3100IS) and ImageZoomBrowser Ex analysis software for Windows. The mycorrhizal type present in each sample was designated according to Harley and Smith (1983) classification. The criteria used in this study for the determination of AM was the presence of vesicles, arbuscules and mycelium at least in one individual root segment and the occurrence of the rest of typical AM structures in the samples (intra or intercellular hyphae, vesicles, coils and arbuscules) and were classified into Arum or Paris-type (Smith and Smith, 1997). The percentage of root length colonized by AM fungi was estimated according to the magnified line-intersect method described by McGonigle et al. (1990).
RESULTS AND DISCUSSION
In this study, Equisetum arvense roots were examined for endomycorhizal association. Results pertaining to diversity of AM fungi and physico-chemical traits of rhizospheric soil are reported in Table 1. The pH, temperature and E conductivity of rhizospheric soil were (6.44± 0.0), (25.5 ±0.0), (15.5± 0.0) respectively. The spore quantification of sample was 20spores/10gm of soil and Glomus rubeforme, Glomus albidum and Glomus species were present in the rhizosphere (Table-1, Plate-1A and H-I). Percent colonization and other endorhizal structures associated with E. arvense L. are shown in Table-2. It was seen that the rhizome is having cent percent total colonization, (Hyphal infection 100%) but mostly the Paris-Type of AM colonization was observed. Arbusculate infection and dark septate endophyte infection (DSE) were observed only on a few segments of rhizome. Whereas Microsclerotium (MS), Microsporangiate like aggregation (MSLA), Cynophycean filamentous like structures (CFLS), Fine endophyte infection (FE) and Peloton like hyphal coils (PLC) were seen in rhizoids which were absent in rhizome part of sporophyte. Hyphal infection was 95% in rhizoids but mostly Intermediate type (IT) i.e. both Arum-type and Paris-type of infections were observed in them. The arbusculate infection was present in 60% of segments whereas vesiculate like infection was observed only in few segments (30%) but true vesicles were not seen in all the segments. More interestingly, the Cynophycean filamentous/trichomes like structures (CFLS) were found in few young rhizoids segments of the sporophyte which indicates that sporophyte may takes nitrogen from cynophycean members which live in symbiotic association with rhizoids (Table-2, Plate- 1B-G). Arbuscular mycorrhizas (AM), as the most widespread type of mycorrhizas, form symbiotic interactions with the roots of 80% of all terrestrial plant species (Read, 1999). Sporophytes of Equicetaceae have been regarded as non-mycorrhizal by several authors as discussed in introduction part of this paper, however, the results in this study and some previous reports as cited earlier indicate that plants may be extensively colonized by AM fungi. Equisetum bogotense collected from Valdivian temperate forest of Patagonia were not obligate mycorrhizal species, but they develop AM under certain conditions, probably related to the habitat and the substrate where they were growing (Fernadez et al., 2008). In spite of the fact that Paris-type is the most common type of AM among the seedless vascular plants (Smith and Smith, 1997; Zhang et al., 2004). Equisetum is commonly associated with damp or wet soils and due to this reason Berch and Kendrick (1982) and Koske et al. (1985) partially attributed their findings of relative paucity of mycorrhizal association. This work represents the record of AM fungi in monilophytes (Equisetum arvensis) of Nongkhyllem Reserve Forest Nongpoh, Meghalaya and constitutes a tread in the study of the importance of AM fungi, endophytes and other microbes in the rhizosphere of this sporophyte. Equisetum plants produce extensive, perennial rhizomatous systems with potentially 80% of their biomass being subterranean. Equisetum species can store abundant starch below ground (Hauke, 1978) and for this reason Equisetum was cultivated for its tubers in the 1920s in upland Bolivia and Peru (Berry 1924). Interactions between Equisetum roots and soil mycoflora may well provide important food sources for burrowing animals and soil invertebrates. Mycorrhizal associations have been considered necessary pre-requisites for plants to colonize terrestrial environments (Pirozynski and Malloch, 1975). In addition to AM, another type of root colonizing fungi called Dark Septate Fungal (DSF), infection has been also reported within the rhizoids of Equisetum sporophyte collected from the said site. Similar Dark Septate Fungal infection in different seedless vascular plants were also reported earlier (Cooper, 1976; Berch and Kendrick, 1982; Dhillion, 1993; Jumpponen and Trappe, 1998). Dark Septate Fungi are defined by Jumpponen (2001) as conidial or sterile fungi that colonize living plant roots without causing any apparent negative effects. The ecology, taxonomic affinities and host range of these DSF are largely unknown (Peterson et al., 2004). Including DSF in mycorrhizal studies would definitely yield valuable information about the importance and diversity of these root colonizers (Jumpponen and Trappe, 1998). The occurrence of Microsclerotia (MS), Microsporangiate like aggregation (MSLA), Cynophycean filamentous like structures (CFLS), Fine endophyte infection (FE) and Peloton like hyphal coils (PLC) in the rhizosphere is especially remarkable, because some structures like Microsporangiate like aggregation, Cynophycean filamentous like structures, Fine endophytic infection and Peloton like hyphal coils have not been previously cited in the literature. As it is becoming important to report in this paper to support the idea of Winter and Friedman (2007) that some seedless vascular plants are capable of forming plant-fungal associations with a diversity of fungal lineages. This information would be very useful to elucidate the nature and ecological importance of these inadequately and un-known root colonizing endophytes (Jumpponen and Trappe, 1998; Jumpponen, 2001; Peterson et al., 2004). This result is in agreement with the finding of Boullard (1957). He suggested that the mycorrhizal symbiosis within seedless vascular plant group ranges from obligate to facultative or non-mycorrhizal. The results also corroborate with the results obtained by Zhao (2000), who studied the mycorrhizal status of 256 species of Lycophytes and Monilophytes and found out that only 16% of them were facultative mycorrhizal.
CONCLUSION
Through this study, it is concluded that Equisetum arvense is an important medicinal spore-dispersing vascular sporophyte used by Khasi tribe in Khasi Hills of Nongkhyllem Reserve Forest, Nongpoh, Meghalaya, India and further depicts that its rhizoids host abundant and diverse endophytic fungal associates that relates the evolution of Equisetum arvense sporophyte from hydric to mesic habitation due to its typical Intermediate type of endophytic and symbiotic association. Also, its endorhizal symbiotic associations may play a significant role in nutrient cycling in the ecosystems and have a broad ecological relevance.
ACKNOWLEDGEMENT
The authors thank Indian Council of forestry Research and Education (ICFRE), an aautonomous council of Ministry of Environment and Forests, Govt. of India, for financial support under the project RFRI- 12/2008-09/SFM and also the Director, RFRI, Jorhat, Assam, India for providing every facility to carry out the experiment.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareEFFECT OF AGE ON CERVICOCEPHALIC KINESTHETIC SENSIBILITY
English4248Ravi Shankar Reddy YEnglish Arun Maiya GEnglish Sharath Kumar RaoEnglishBackground: The ageing process is important factor related to proprioception. Age-related reduction in
proprioception in weight-bearing joints, such as the knee or ankle joint, of asymptomatic adults has been
documented extensively. However, no age related increases in cervical repositioning error were reported in asymptomatic adults
Objective: The aim of the study was to determine the effect of age on cervical position sense
Materials and methods: 150 asymptomatic young adults (age range, 20-60 years) were recruited in to the study. The subjects were divided in to four groups i.e. 20-29 years, 30-39 years, 40-49 years and 50- 60 years. The subjects were measured for reposition errors (degrees) by the Cervicocephalic kinesthetic sensibility tests, which include Head-to-Neutral Head Position (NHP) repositioning tests and Head-to- Target repositioning tests with Cervical Range of Motion (CROM) Device. The two repositioning tests were performed in the sagital, transverse, and frontal planes.
Results: The results of one way ANOVA showed there is significant difference between the groups
(PEnglishNeck proprioception; Neck repositioning test; AgingINTRODUCTION
Proprioception is a term commonly used to describe the ascending information by the afferent receptors towards the central nervous system contributing to the neuromuscular control of movement and encompasses the sensation of joint movement (kinesthesia) and joint position (joint position sense).1,2 It has been demonstrated that proper function of the head– neck system relies on proprioceptive information provided from receptors in the zygapophyseal joints and small intrinsic muscles.3 Proprioception, including joint positioning sense, protects the joint by regulating joint stiffness through the activation of mechanoreceptors and the muscle spindle system.4,5 The cervicocephalic kinesthetic sensibility test (a frequently adapted method of determining neck proprioception) was been used to examine each subject‘s ability to return their head to a predetermined position without visual cues after they have moved.6 The common methods used in literature to assess cervical proprioception include cervical range of motion (CROM) device, electromagnetic tracking system (FASTRAK), Rod and frame test, ultrasound based coordinate measuring system (CMS 70P), CA6000 spinal motion analyzer, laser pointer method.7-12 It is well established that as the age increases degenerative changes occurs in the cervical column and it is well established that motion of the cervical spine decreases with age, most probably because of the development of degenerative changes.13 As a part of the degeneration process there might be an altered proprioceptive input with increasing age.14,15 Age-related reduction in proprioception in weight-bearing joints such as the knee or ankle joint, of asymptomatic adults has been documented extensively and proprioceptive deficits have been associated with chronic pain, injury and muscle fatigue.16,17 However, no age related altered proprioceptive inputs are reported in cervical spine in asymptomatic adults. So the purpose of the study is to determine if there is a positive correlation between altered proprioception inputs with increasing age in asymptomatic individuals.
METHODS
Subjects:
Advertisements in the University, physical therapy department, and local city were given in the form of posters and lectures for voluntary participation of the subjects. The study included 150 asymptomatic subjects (range, 20-70 years). All subjects reported that they had no neck pain at the time of the study. To be considered asymptomatic, a subject could not have had any previous treatment for neck pain, and no current neck pain. Exclusion criteria included are traumatic spinal injury, whiplash associated disorder, central nervous system impairment, demonstrated by parasthesia, vestibular impairment demonstrated by vertigo, dizziness, or motor imbalance, neck pain induced by cervical motion in the range tested for the study. Subjects were required to attend two sessions. The first was to familiarize them with the equipment and repositioning tasks. Ethical approval for this study was granted by the University Ethics Committee. All participants signed a written consent form prior to participating in the experiment.
Instrumentation:
Cervical Range of Motion (CROM) device:
The cervical range of motion (CROM) device is a type of goniometer designed specifically for the cervical spine and was used to measure cervical range of motion.18 The Cervical Rangeof-Motion Device (CROM) has been evaluated most often, with 7 studies assessing its reliability on healthy volunteers or symptomatic patients.19 The CROM has 3 inclinometers, one to measure in each plane, and is strapped to the head. One gravity dial meter measures flexion and extension, another gravity dial meter measures lateral flexion and a compass meter measures rotation with its accuracy reinforced by 2 magnets placed over the subject's shoulders. CROM device is effectively used in clinical set up, Easy to apply and Cost effective. CROM device has good Criterion validity (r = 0.89 – 0.99) and Reliability (ICC= 0.92 - 0.96).18-20
Measurement of cervicocephalic kinesthetic sensibility
The subjects were asked to sit upright in a comfortable position and look straight ahead to be determined as the neutral head position (NHP). The CROM unit was placed on top of the head and attached posteriorly using the Velcro strap. The magnetic part of the unit was then placed so that it sat squarely over the shoulders. The investigator calibrated the CROM device to a neutral head position.
For the cervicocephalic kinaesthetic sensibility tests, subjects were required to keep the head in the NHP and were told to close their eyes throughout the subsequent tests. The first test was Head-to-Neutral Head Position (NHP) repositioning test.21 The subjects were instructed to turn the head fully to the left and back to what they considered the starting point in a controlled fashion without opening their eyes. When the subjects reached the reference position the subject‘s relocation accuracy was measured in degrees with the CROM device. In the second repositioning test is Head-to-Target repositioning tests.22 The investigator moved the subject‘s head slowly to the predetermined target position, 65% of maximum range of motion. The speed of passive neck motion was very slow as higher speeds have been associated with significant differences in vestibular function according to age.23 The head was maintained in the target position for 3 seconds and the subject was asked to remember that position and the head was brought to neutral position and then the subject were asked to reposition actively by moving the head to the target position. When the subjects reached the reference position, the subject‘s relocation accuracy was measured in degrees with CROM device. The two repositioning tests were performed in the sagital, transverse, and frontal planes. Each test position was measured three times and the average of the three was taken for analysis.
Statistical Analysis
One way ANOVA was used to compare joint reposition errors within the groups. The statistical analysis was done using the SPSS 11.0 for windows software. The statistical significance value will be set at 0.05 with 95% confidence interval and a p value less than or equal to 0.05 will be considered to be significant.
RESULTS
The mean values of repositioning errors during cervicocephalic kinesthetic sensibility testing in Neutral Head position (table 1) and target reposition (table 2) shows greater increase in repositioning errors as the age increases. The results of one way ANOVA showed there is significant difference between the groups (PEnglishhttp://ijcrr.com/abstract.php?article_id=2159http://ijcrr.com/article_html.php?did=21591. Strimpakosa N, Sakellarib V, Gioftsosb G, Cervical joint position sense: an intra- and inter-examiner reliability study. Gait and Posture 2006;23:22-31
2. Lephart SM, Fu FH. Proprioception and neuromuscular control in joint stability. Champaign: Human Kinetics; 2000
3. Cordo P, Gurfinkel VS, Bevan L, Kerr GK. Proprioceptive consequences of tendon vibration during movement. Journal of Neurophysiology 1995;74:1675-88.
4. Johansson H, Djupsjobacka M, Sjolander P. Influences on the gammamuscle spindle system from muscle afferents stimulated by KCl and lactic acid. Neuroscience Research 1993;16:49-57.
5. Solomonow M, Zhou BH, Harris M, Lu Y, Baratta RV. The ligamento-muscular stabilizing system of the spine. Spine 1998;23:2552-62.
6. Leea H, Wanga J, Yaoc G, Association between cervicocephalic kinesthetic sensibility and frequency of subclinical neck pain. Manual therapy 2007:1-7.
7. Dall‘alba P, Sterling M, Trelaeven J, Jull G. Cervical range of motion discriminates between asymptomatic and whiplash subjects. Spine 2001;26(19):2090–94
8. Treleaven J, Jull G, Sterling M. Dizziness and unsteadiness following whiplash injury: characteristic features and relationship with cervical joint position error. Journal of Rehabilitation Medicine 2003;35(1):36–43.
9. Kristjansson E, Hardardottir L, Asmundardottir M, Gudmundson K. A new clinical test for cervicocephalic kinesthetic
sensibility: ??the fly‘‘. Archives of Physical Medicine and Rehabilitation 2004;85(3): 490–5.
10. Jasiewicza JM, Treleavenb J, Condiea P, Jullb G Technical and measurement report Wireless orientation sensors: Their suitability to measure head movement for neck pain assessment Manual Therapy 2007;12: 380-385
11. Janice K, Ruhul M, Field E, Ability to reproduce head position after Whiplash Injury. Spine 1997;22:865-868.
12. Jordan K. Reliability Studies for Cervical Spine ROM Journal of Manipulative and Physiological Therapeutics 2000;23:180- 195.
13. Kristjansson E, Dall‘Alba P, Jull G. A study of five cervicocephalic relocation tests in three different subject groups. Clin Rehabil 2003;17:768-74.
14. Tenga C, Chaia H, Laib D Cervicocephalic kinesthetic sensibility in young and middleaged adults with or without a history of mild neck pain Manual Therapy 2007;12:22-28
15. Skinner HB, Barrack RL, Cook SD. Agerelated decline in proprioception. Clinical Orthopaedics and Related Research 1984:208-11.
16. Gilsing MG, Van den Bosch CG, Lee SG, Ashton-Miller JA, Alexander NB, Schultz AB, et al. Association of age with the threshold for detecting ankle inversion and eversion in upright stance. Age Ageing 1995;24:58-66.
17. Robbins S, Waked E, McClaran J. Proprioception and stability: foot position awareness as a function of age and footwear. Age Ageing 1995;24:67–72
18. Michel T, Cecile S, Anne-Marie B. Criterion valisity of Cervical Range of Motion (CROM) device for Rotational range of motion on healthy adults: Journal of Orthopedic and Sports Physical Therapy 2006:36:242-248.
19. Tousignant M, de Bellefeuille L, O'Donoughue S, Grahovac S. Criterion validity of the cervical range of motion (CROM) goniometer for cervical flexion and extension. Spine. 2000;25:324-330.
20. Tousignant M, Duclos E, Lafleche S, et al. Validity study for the cervical range of motion device used for lateral flexion in patients with neck pain. Spine. 2002;27:812- 817.
21. Lee H-Y, Teng C-C, Chai H-M, Wang S-F. Test-retest reliability of cervicocephalic kinesthetic sensibility in three cardinal planes. Manual Therapy 2006;11:61-8.
22. Loudon JK, Ruhl M, Field E. Ability to reproduce head position after whiplash injury. Spine 1997;22:865-8.
23. Goebel JA, Hanson JM, Fishel DG. Agerelated modulation of the vestibulo-ocular reflex using real and imaginary targets. Journal of Vestibular Research 1994;4:269- 75.
24. Gilman S. Joint position sense and vibration sense: anatomical organization and assessment. J Neurol Neurosurg Psychiatry. 2002 Nov;73(5):473-7
25. Richmond FJR, Bakker DA. Anatomical organization and sensory receptor content of soft tissues surrounding upper cervical vertebrae in the cat. Journal of neuro Physiology 1982; 1:49-61.
26. David T, Douglas G, Michael C. Nevitt. The effects of impaired joint position sense on the development and progression of pain and structural damage in knee osteoarthritis. Arthritis Rheum 2009;15:1070-1076.
27. Ulrike V, Stefanie H, Friso , William D. Reproducibility of postural control measurement during unstable sitting in low back pain patients. BM
sensibility: ??
-the fly‘". Archives of Physical Medicine and Rehabilitation 2004;85(3): 490-5.
10. Jasiewicza JM, Treleavenb J, Condiea P, Jullb G Technical and measurement report Wireless orientation sensors: Their suitability to measure head movement for neck pain assessment Manual Therapy 2007;12: 380-385
11. Janice K, Ruhul M, Field E, Ability to reproduce head position after Whiplash Injury. Spine 1997;22:865-868.
12. Jordan K. Reliability Studies for Cervical Spine ROM Journal of Manipulative and Physiological Therapeutics 2000;23:180- 195.
13. Kristjansson E, Dall‘Alba P, Jull G. A study of five cervicocephalic relocation tests in three different subject groups. Clin Rehabil 2003;17:768-74.
14. Tenga C, Chaia H, Laib D Cervicocephalic kinesthetic sensibility in young and middleaged adults with or without a history of mild neck pain Manual Therapy 2007;12:22-28
15. Skinner HB, Barrack RL, Cook SD. Agerelated decline in proprioception. Clinical Orthopaedics and Related Research 1984:208-11.
16. Gilsing MG, Van den Bosch CG, Lee SG, Ashton-Miller JA, Alexander NB, Schultz AB, et al. Association of age with the threshold for detecting ankle inversion and eversion in upright stance. Age Ageing 1995;24:58-66.
17. Robbins S, Waked E, McClaran J. Proprioception and stability: foot position awareness as a function of age and footwear. Age Ageing 1995;24:6772
18. Michel T, Cecile S, Anne-Marie B. Criterion valisity of Cervical Range of Motion (CROM) device for Rotational range of motion on healthy adults: Journal of Orthopedic and Sports Physical Therapy 2006:36:242-248.
19. Tousignant M, de Bellefeuille L, O'Donoughue S, Grahovac S. Criterion validity of the cervical range of motion (CROM) goniometer for cervical flexion and extension. Spine. 2000;25:324-330.
20. Tousignant M, Duclos E, Lafleche S, et al. Validity study for the cervical range of motion device used for lateral flexion in patients with neck pain. Spine. 2002;27:812- 817.
21. Lee H-Y, Teng C-C, Chai H-M, Wang S-F. Test-retest reliability of cervicocephalic kinesthetic sensibility in three cardinal planes. Manual Therapy 2006;11:61-8.
22. Loudon JK, Ruhl M, Field E. Ability to reproduce head position after whiplash injury. Spine 1997;22:865-8.
23. Goebel JA, Hanson JM, Fishel DG. Agerelated modulation of the vestibulo-ocular reflex using real and imaginary targets. Journal of Vestibular Research 1994;4:269- 75.
24. Gilman S. Joint position sense and vibration sense: anatomical organization and assessment. J Neurol Neurosurg Psychiatry. 2002 Nov;73(5):473-7
25. Richmond FJR, Bakker DA. Anatomical organization and sensory receptor content of soft tissues surrounding upper cervical vertebrae in the cat. Journal of neuro Physiology 1982; 1:49-61.
26. David T, Douglas G, Michael C. Nevitt. The effects of impaired joint position sense on the development and progression of pain and structural damage in knee osteoarthritis. Arthritis Rheum 2009;15:1070-1076.
27. Ulrike V, Stefanie H, Friso , William D. Reproducibility of postural control measurement during unstable sitting in low back pain patients. BM
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcarePRE-FORMULATION ASSESSMENT: FRUIT OF CUMINUMCYMINUM, LINN.
English4957Shanthi.AEnglish R. RadhaEnglishCuminumcyminum, (Linn.)Correa; belonging to the familyUmbelliferaeis commonly known as Cumin (Eng.) and Jira (Hindi). It is a glabrous, annual herb. Cultivated as a cold season crop on the plains and as summer crop on the hills in Northern India, a native of west Asia. It has the traditional value for curing various ailments like Asthma, fever, skin diseases, leprosy and also as a helminthiasis. This traditionally useful part (fruit) was standardized based on the pharmacognostic, physico-chemical and chromatographic conditions. Quality assessment by determining the limits of Microbials, heavy metals, pesticides and Aflatoxin as per WHO guidelines. The report states that the sample taken was standard under AyurvedicPharmacopoeial limits and WHO. The preliminary phytochemical screening confirms the presence of Essential oils, Glycosides, sterols and proteins. HPTLC finger print states the presence ofN nine different constituents and determination of total volatile oil content by Gas chromatography was found to be about 0.887%w/w. From this the report confirms that the sample was standard enough to use in the developing a herbal formulation.
EnglishCuminumcyminum, standardization, phytochemical screening, chromatographic identification and total volatile oil estimation by GCINTRODUCTION
Herb is a plant that is valued for flavor, scent, medicinal or other qualities other than its food value1 . Over the last few years, researchers have aimed at identifying and validating plant derived substances for the treatment of various diseases. Interestingly, it is estimated that more than 25% of modern medicines are directly or indirectly derived from plants2 . With the emerging interest in the world to adopt and study the traditional system and to exploit their potentials based on different health care systems, the evaluation of the rich heritage of the traditional medicine is essential3 .The general standardization protocols to determine the percentage of active medicaments could not be followed for Ayurvedic herbal preparations. The procedures have to be modified in order to make the preparation safe. The approach has to be made from raw materials to finished products evaluation for the successful outcome. Which include the standardization of rawmaterial. The drugs of plant origin especially of herbaceous nature are identified with their origin, common name, scientific nomenature,family, geographical source, cultivation, collection, preservation, storage, macroscopy,microscopy, chemical composition, identity, purity, strength and assay, substitute andadulterants etc., The microscopic examination of root which includes Transverse sectionand Longitudinal section are made for identification4 . Cuminumcyminum is commonlyknown as ?Cumin‘ cultivated as a cold season crop on the plains and as summer crop on the hills in Northern India, a native of west Asia and Cultivated throughout India. Cumin is one of the constituents of siddha preparation Attalicuranam5 . The ripe fruits have been used traditionally for curing various ailments like Asthma, fever, skin diseases, leprosy and also as a helminthiasis6 . And it also used in neurological disorders7 , anti-depressant effects8 . The present work was based on the standardization of Cuminumcyminum fruit as per WHO guidelines9 . The complete standardization of this variety may be used for formulation development in future.
EXPERIMENTAL PART
1) Chemicals and reference drugs:
All the chemicals used in this present work were analytical grade in nature. The chromatographic estimations were done in the Asthagiri Herbal Research Foundation (Chennai) are chromatographic grade.
2) Collection of plant materials:
The roots of Cuminumcyminum were provided by M/S AnnaiAravindHerbals,Chennai. The sample was taken for authentication.
3) Macroscopic evaluation:
Organoleptic characters like colour, odour, taste, size, shape and other characters can be identified entirely or it fragment. This is authentication of crude drug with its genuine varierty10 .
4) Microscopic evaluation:
The microscopic appearance of the drug both in section view and in powdered form for its authentication with its genuine variety was performed11 .
5) Determination of foreign matter:
About 100g of the drug sample was spread out on a thin layer. The foreign matter was detected by inspection, separated and weighed. And the percentage of foreign matter was calculated12 .
6) Determination of total ash:
About 2g accurately weighed drug was incinerate in a tarred silica dish at a temperature not exceeding 4500 c until free form carbon, cooled and weighed. The percentage of ash obtained was calculated with reference to the air-dried drug13 .
7) Determination of acid-insoluble ash:
Total ash obtained boiled for 5 minute with 25ml of dilute hydrochloric acid; insoluble matter was collected in a Gooch crucible, or an ash less filter paper, again washed with hot water and ignited to constant weight. The percentage of acid-insoluble ash was calculated with reference to the air dried drug.
8) Determination of water soluble ash:
The ash obtained was boiled for 5 minutes with 25ml of water; insoluble matter was collected in a Gooch crucible or on an ash less filter paper, wash with hot water, and ignited for 15minutes at a temperature not exceeding 4500 c. The weight of the insoluble matter obtained was subtracted from the weight of the ash; the different in the weight represents the watersoluble ash. The percentage of water-soluble ash was calculated with reference to the air-dried drug.
9) Determination of alcohol soluble extractive:
About 5g of the air-dried drug was coarsely powdered and macerated with 100ml of Alcohol (specified strength) in a closed flask for 24hours, shake frequently during 6hours and allowed to stand for 18hours. Filtered rapidly, taking precautions against loss of solvent and evaporated 25ml of the filtrate to dryness in a tarred flat bottomed shallow dish, and dried at 1050C to constant weight and weighed. The percentage of alcohol soluble extractive was calculated with reference to the air-dried drug.
10) Determination of water soluble extractive:
About 5g of the air-dried drug was coarsely powdered and macerated with 100ml of Chloroform water in a closed flask for 24hours, shake frequently during 6hours and allowed to stand for 18hours. Filtered rapidly, taking precautions against loss of solvent and evaporated 25ml of the filtrate to dryness in a tarred flat bottomed shallow dish, and dried at 1050C to constant weight and weighed. The percentage of alcohol soluble extractive was calculated with reference to the air-dried drug.
11) Determination of moisture content (loss on drying)
About 10 g of the drug accurately weighed and place in a tarred evaporating dish and dried at 1050 c for 5 hours, and weighed. The drying and continued and weighed at 1hour interval until difference between two successive weighing corresponds to not more than 0.25 percentage is achieved.
12) Quality parameters:
Limit test for microbial limits: Microbial limits were detected for Total aerobic count, Total bacterial count, Total Yeast and moulds, Test for Escherichia Coli, Salmonella species, Psueudomonasaerugenosa, Staphyllococusaureus and Aflatoxins14 .
13) Pesticide residue:
A pesticide is any substance or mixture of substance intended for preventing, destroying or controlling any pest. Chromatographic estimation can be used for detecting and quantifying the pesticide limits in the crude drug was detected15 .
14) Determination of heavy metals:
The test for heavy metals is designed to determine the content of metallic impurities that are coloured by Sulphide ion, under specified conditions. The limit for heavy metals is indicated in the individual monograph in terms of parts of lead per million of substance (by weight). It includes the limit test for Arsenic, Lead, Mercury, and Cadmium16 .
15) Preliminary phytochemical screening:
Preliminary phytochemical screening for alkaloids, glycosides, tannins, terpenoids, phenols, sterols, proteins, amino acids, volatile oils, flavonoids, saponins, fixed oils, essential oils, coumarins, sugars and others17
16) Study of colour change under UV light:
The powdered drug was studied for any colour change under UV light. The sample was divided into three parts and each was studied under ordinary light, short UV (254nm) and long UV (356nm). First part is observed as such, second part observed after treating with 50% Hydrochloride acid, and third part observed after treating with 50% Sodium hydroxide18 .
17) Chemical identity: TLC analysis of plant drugs19
. About 4g of the sample was soaked in chloroform for 18 hours, boiled, filtered and concentrated to 10ml in standard flak. 20Pl of the sample was applied on the Merck aluminium plate pre-coated with silicagel 60 F254 of 0.2mm thickness and the plate was developed in water: Aceto nitrile: methanol: ethyl acetate: hexane (1.5:5.0:0.5:1.5:1.5). The plate was dried and photographed under UV 254 and 366nm.
HPTLC fingerprint:
A Cammag HPTLC system equipped with a sample applicator Linomat IV, twin trough plate development chamber, TLC SCANNER II and Wincats used as an integration software 4.02 (Switzerland). TLC aluminium plates pre-coated with silica gel 60 F 254 (10x10cm, 0.2mm thick) were used. The estimation has been done using the following chromatographic conditions. The plate was dried and scanned at 200 -450nmusing Deuterium lamp the finger prints was developed.
18) Quantitative estimation:
Volatile oil determination was carried out by Gas chromatography. It is an excellent tool for the separation, characterization and quantitative estimation of volatile component of essential oil containing drugs. The identity of the components was assigned by comparing their GC retention times with those of authentic samples, as well as of the components of other essential oils. GC was performed on a Varian gas chromatograph, model cx-3400, under the following conditions: carrier gas, hydrogen; injector and detector temperatures, 2208C and 2258C, respectively; using a capillary column (Supelcowax-10, 30 m _ 0.3 mm), with oven temperature programmed from 808C at 58C/min to 1508C, then at 78C/min to 2158C20 .
RESULTS
1. Authentication of the plant material:
Plant part used was authenticated by Prof. P. Jayaraman, National institute of herbal sciences, West Tambaram, Chennai. The voucher no: PARC/2010/660.
2. Determination of foreign matter:
Foreign matters were present in less than one percentage (Limit NMT 2%) the value was obtained from the triplicate of the analysis.
3. Macroscopic evaluation:
Type: Cremocarp.
Colour: brown with light coloured ridges.
Odour:Umbelliferous characteristic.
Taste: Spicy. Size: 4-6 m long, 2 mm wide.
Shape: Ellipsoidal, elongated, tapering at both ends.
Extra features: Lateral: Slightly compressed.
Ridges: 5 primary and 4 secondary ridges. The macroscopic image was given in Fig 1: Cuminumcyminum fruit.
4. Microscopic evaluation:
a. Transverse section:
It shows Outer epidermis: polygonal cells
Testa: brown coloured polygonal cells, bundles of lignified sclerenchymatous fibres
Inner epidermis: polygonal cell - regularly arranged.
Mesocarp: few layer of parenchyma with five vascular bundles - primary ridges and 6 vittaes under secondary ridges (4dorsal side, 2 on commissural surface).
Endocarp: polygonal cells with fixed oils, aleurone grains and rosette of calcium oxalate crystals, fibres in carpophore. Fig 2: Transverse section of Cuminumcyminum fruit. b. Powder:
Colour - Brown
Endosperm cells -Aleurone grains, fixed oil, calcium oxalate crystals.
Fragments of testa: brown polygonal cells.
Mesocarp: Fibrovascular elements, thick walled sclerenchymatous cells and large oil ducts.
Vessels - annular spiral thickening
Fig 3: Microscopy ofCuminumcyminum fruit.
5. Physico-chemical constants
The table No: 1 shows that the mean valueof physical constants such as LOD, Ash values and extractive values. The report shows the presence of above values within the limits prescribed under Ayurvedic Pharmacopoeia of India, and the sample contain high proportion of water and alcohol soluble constituents.
6. Microbial determination:
The table No: 2 shows the presence of microbial limits of the sample. And the report shows that as per the WHO standards, the plant material is free from microbial load and ranges within the standard limit and the Aflatoxin. And it was found to be safe for further use in formulation development.
7. Determination of pesticide
Pesticide analysis of the raw material states that the DDT, Benzene hexa chloride, Aldrin, Dialdrin, Lindane, Chloropurophos and Enoculphan type of pesticides are not detected in the drug sample. ND - not detected (concentration less than the minimum detection limit even in ng/l units). From the above results it can be concluded that the plant material is totally safe and there is no traceable limit of pesticide in them.
8. Quality parameters:
The table No: 3 shows the presence of heavy metal limits of the sample. The report shows that the raw material is free from Arsenic, Cadmium and Mercury and Lead is present within the standard limits.
9. Preliminary phytochemical screening:
Preliminary phytochemical screening indicates the presence of Essential oils, Glycosides, sterols and proteins.
10. Study of colour change under UV light:
The table No: 4 represents the colour change of the sample under normal, long and short UV. Colour change was observed under ordinary light with acid and alkali medium with respect to powder alone and in short UV only in alkaline medium the colour change was observed and there was no change in colour was observed under long UVregion.
11. Chemical identity:
The table No. 5 indicates the Rf values obtained for the sample and the number of constituents identified with that respective Rf values. The figure 3 gives the TLC image and the figure 4 gives the HPTLC finger print data. The result obtained from the HPTLC studies showed that, therewere nine spots were observed.
12.Quantification
: The total volatile oil content of the Cumimuncyminumfruit estimated by Gas Chromatography method was found to be about 0.887% w/w.
DISCUSSION
The macroscopic and the microscopic evaluation of drug establishing its quality control profile and according to WHO, botanical standardswere proposed as a protocol for the diagnosis. The physic-chemical constants were lies within the AyurvedicPharmacopoeial limits states that the sample is free from adulterants and the quality evaluations for microbial, heavy metals states that they were present with in the WHO limits and the preliminary phytochemical evaluation states the presence of secondary metabolites and the HPTLC finger print reveals the number of constituents present and the quantification report states the purity of the sample based on the quantity of phytoconstituents.
CONCLUSION
The reports states that the Cumimuncyminumwas authenticated based on its pharmacognostic, physico-chemical and phytochemical aspect. And the qualitative estimation impliessafety of the sample due to the absence of heavy metals, pesticides and microbes. Quantitative estimation states the therapeutically useful constituent (volatile oil) is present in it.Thus the Cumimuncyminum is valid for its therapeutic potency.And thus it can be used in the herbal formulation development.
ACKNOWLEDGEMENT
The authors would like to express their gratitude to the supporters those who are directly and indirectly participated in presenting this work for publication and also supported for carrying out the above research work.And special thanks to Dr. A. Kannan, Asthagiri Herbal Research Foundation for his timely help in completing the HPTLC analysis. Funding source: Nil.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30General SciencesPILOT SCALE PRODUCTION OF BIODIESEL FROM MACRO ALGAE COLLECTED FROM COROMANDAL
COAST, EAST COAST INDIA
English5876Ramganesh SelvarajanEnglish Senthil KumarEnglish Rangasamy MurugesanEnglish Sanniyasi ElumalaiEnglishBioenergy is one of the most important components to mitigate greenhouse gas emissions and substitute of fossil fuels. The need of energy is increasing continuously, because of increases in industrialization and population. The basic sources of this energy are petroleum, natural gas, coal, hydro and nuclear. The ability of algae to fix Co2 can also be an interesting method of removing gases from power plants, and thus can be used to reduce greenhouse gases with a higher production algal biomass and consequently higher biodiesel yield. Present research such as biodiesel from macroalgae having species Sargassum ilicifolium and Turbinaria ornata. Biodiesel production (methyl ester) was found maximum in Sargassum ilicifolium (8.01%) and minimum in Turbinaria ornate (7.03%).More over sediment (glycerine, pigments and other elements) were higher in Turbinaria ornate(4.26g) than in Sargassum ilicifolium(2.93g). It was observed that Turbinaria ornata could produce large amount of poly unsaturated fatty acids (PUFA) as the major storage lipid which accounted 80.44% than the Sargassum ilicifolium is about 75.98% respectively. Since intracellular lipids can be enhanced by controlling the conditions such as nutrient limitation, limitation of light and salt stress, it can be expected that the TAG yield of macro algae can further be increased by the same approaches. This may be a First report in India that the biodiesel producing macroalgae, their detailed ultrastructure and the lipid droplets storage. If we go further to demonstrate this model in the coastal India, there will be a larger opportunities to produce cost effective biofuel or
biodiesel and glycerine production as well as the rural employment creation.
EnglishINTRODUCTION
Biodiesel is defined as the monoalkyl esters of long chain fatty acids, an alternative for fossil fuel. The most common biodiesel constituent used today is fatty acid methyl esters. Bioenergy is one of the most important components to mitigate greenhouse gas emissions and substitute of fossil fuels. The need of energy is increasing continuously, because of increases in industrialization and population. The basic sources of this energy are petroleum, natural gas, coal, hydro and nuclear. Biodiesel is non toxic completely biodegradable fuel with reduced sulfur emission. The release of sulfur content and carbon monoxide would be cut down by 30% and 10%, respectively, by using biodiesel as energy source. Using biodiesel as energy source, the gas generated during combustion could be reduced, and the decrease in carbon monoxide is owing to the relatively high oxygen content in biodiesel. Biodiesel (monoalkyl esters) is one of such alternative fuel, which is obtained by the transesterification of triglyceride oil with monohydric alcohols. It has been well-reported that biodiesel obtained from canola and soybean, palm, sunflower oil, algal oil as a diesel fuel substitute. The burning of an enormous amount of fossil fuel has increased the Co2level in the atmosphere, causing global warming. Biomass has been focused on as an alternative energy source, since it is a renewable resource and it fixes Co2in the atmosphere through photosynthesis. If biomass is grown in a sustained way, its combustion has no impact on the Co2 balance in the atmosphere, because the Co2 emitted by the burning of biomass is offset by the Co2 fixed by photosynthesis [Macedo]. Among biomass, algae (macro and microalgae) usually have a higher photosynthetic efficiency than other biomass. The ability of algae to fix Co2 can also be an interesting method of removing gases from power plants, and thus can be used to reduce greenhouse gases with a higher production algal biomass and consequently higher biodiesel yield. Micro-algae have received so far more attention with respect to macro-algae as agents for enhanced Co2 fixation due to their facile adaptability to grow in ponds or bioreactors and the extended knowledge on several strains used for fish feeding. Macro-algae are extensively grown and used as food in Asiatic Countries, or as source of chemicals. They are usually collected from natural water basins where they are seasonally available. Only recently they have been considered for energy production, and the potential of some Pacific Ocean strains has been preliminarily studied. In fact algae are the highest yielding feedstock for biodiesel. It can produce up to 250 times the amount of oil per acre as soybeans. In fact, producing biodiesel from algae may be only the way to produce enough automotive fuel to replace current gasoline usage. Algae produce 7 to 31 time greater oil than palm oil. It is very simple to extract oil from algae. The idea of using algae as a source of fuel is not new but it is now being taken seriously because of the escalating price of petroleum and, more significantly, the emerging concern about global warming that is associated with burning fossil fuels. No literature is found regarding present research such as biodiesel from macroalgae having species Sargassum ilicifolium and Turbinaria ornata. Therefore the present work will focus on existing gap between the fossil fuel and Microalgae based Biofuel.
MATERIALS AND METHODS
Study Site
The Present experiment was carried out in the research lab of Department of Plant Biology and Biotechnology, Presidency College (Autonomous), Chennai, India and Sopisticated Analytical Instrumentation Facility (SAIF) Indian Institute of Technology Madras (IITM), Chennai, India.
Macro algae Sample collection
Two different Algae (Sargassum ilicifolium and Turbinara ornata) were collected from the village Vadakadu, Rameshwaram - Taluk, Ramanathapuram, Tamil Nadu, India. It is located at 9.28°N 79.3E. It has an average elevation of 10 meters (32 feet).
Anatomical studies of the Macrolgae:
The Collected Macrolgae was fixed in FAA (Formalin-5ml + Acetic acid-5ml + 70% Ethanol-90ml) for two hours. The materials were washed in distilled water and dehydrated through graded series of Tertiary butyl alcohol (Sass, 1940). Following dehydration, the materials were infiltrated with paraffin wax controlled temperature (55°C). After infiltration, the specimens were cast into paraffin blocks and the blocks were stored in refrigerator for sectioning. Serial sections to the thickness of 6-8μm were prepared with the help of rotary Microtome. The sections were dewaxed and stained with 0.05% Toluidine blue O (O‘Brien et al., 1964) (dissolved in water) for general anatomical studies. Since it is a Meta chromatic dye, it gave good results for studying gross anatomical features of the inner parts.
Nile blue Staining:
For localization of the Lipid Bodies in the Algae, sections were stained with Nile Blue Stain. The sample or frozen sections are fixated in formaldehyde, then immersed for 20 minutes in the Nile blue solution and rinsed with water. For better differentiation, it is dipped in 1% acetic acid for 10–20 minutes until the colors are pure. This might take only 1–2 minutes. Then the sample is thoroughly rinsed in water (for one to two hours). Afterwards, the stained specimen is taken on a microscope slide and excess water is removed. The sample can be embedded in glycerol or glycerol gelatin. Both external and microtome sections were photographed with NIKON Coolpix-8400 Digital camera and NIKON Labphoto-2 microscopes. Magnifications of the micrographs are shown by the scale-bars.
Lipid Extraction and Oil Conversion from Macroalgae
Macroalgae were ground with motor and pestle as much as possible. The ground algae were dried for 20 min at 80°C in a incubator for releasing water. The yield of lipid contents were compared using different solvent system such as, organic solvents (1-butanol, ethanol, n-hexane / Ether), catalyst (NaOH), and different levels of feeding frequency, pH value, temperature, and reaction time. Hexane and ether solution (1:1) were mixed with the dried ground algae and amber colored oil extracts were collected on top of the solution Then the mixture was kept for 24 h for settling.
Biomass collection
The Macroalgal biomass was collected after filtration and weighted.
Evaporation
The extracted oil was evaporated in vacuum to release hexane and ether solutions using rotary evaporator.
Transesterification
Transesterification is the process of exchanging the organic group R" of an ester with the organic group R' of an alcohol. These reactions are often catalyzed by the addition of an acid or base catalyst. The reaction process is called transesterification
Mixing of catalyst and methanol
NaOH (0.25 g) was mixed with methanol (24 mL) and stirred properly for 20 min. The conical flask containing solution was shaken for 3 h by electric shaker at 300rpm.
Settling
After the manual shaking the biomass solution was kept for 16 h to settle the biodiesel and sediment layers clearly.
Separation of biodiesel
The biodiesel was separated from sedimentation by separating funnel carefully. Quantity sediment (glycerin, pigments, etc.) was measured.
Washing
The collected Macroalgal biodiesel was washed by 5% water until it become clean.
Storage
Biodiesel production was measured by using measuring cylinder; pH was measured and stored at room temperature for further analysis.
Gas Chromatography and Mass Spectroscopic Studies
The collected Macroalgal biodiesel from sample was processed with GC and MS (JEOL GC mate). Lipid fraction was resuspended in nhexane and applied to silica gel column chromatography. Aliphatic hydrocarbon fraction passes through the column fatty acid and carotenoid fractions were trapped. Passing through fraction was defined as hydrocarbon fraction, lipid components in hydrocarbon fraction were identified by GC/MS. The sample (1µl) was evaporated in a split less injector at 300°C. The results were recorded and compared with the Fosssil diesel and gasoline oils. The methyl esters of fatty acids were quantified by a gas chromatograph (Agilant-JEOL GC and MS). The column (HP5) was fused silica 50m x 0.25 mm I.D. Analysis conditions were 20 minutes at 100°C the 3°/ min to 235°C for column temperature, 240°C for injector temperature, helium was the carrier gas. The weight percentages of fatty acids were approximated by the area of the detector response. The fatty acid methyl esters were identified by gas chromatography coupled with mass spectrometry.
Fourier Transform Infra-Red Spectrometry
In our study Perkin Elmer model spectrum-I PC was used. FT - IR spectra (Resolution: 4 cm-1, Scan Number: 3) were performed after evaporation of the Lipid fraction on the Thalium bromide tablets.
RESULTS AND DISCUSSION
Percent Dry weight of algae before oil extraction was lower in Sargassum ilicifolium than in Turbinaria ornata (Table 1). However Extracted oil was higher in Sargassum ilicifolium than in Turbinaria ornata. However biomass after oil extraction was found maximum in Turbinaria ornata than the Sargassum ilicifolium (Table 1). Biodiesel production (methyl ester) was found maximum in Sargassum ilicifolium and minimum in Turbinaria ornata.More over sediment (glycerine, pigments and other elements) were higher in Turbinaria ornata than in Sargassum ilicifolium (Table 1). There is no significant difference in pH between both species. In (Fig 5) photograph shows esterification (biodiesel and residual layer) and biodiesel.
Sargassum ilicifolium The Macroalgal body consist of flat leaf like lateral appendages central thick stipe. The stipe or stem has an epidermal layer of squarish cells, wide, thin walled, less compact cortical tissues and central core of thick walled angular medulla.
Sections stained with Nile Blue
Epidermal cells stain dark and Cortical cells and medullary cells stain purple. Darkly stained Lipid bodies are seen in the cells of the medulla. (Fig. 1.1, 1.2.)
Sections stained with Toludine Blue O
The epidermal cells of macroalgal stain dark. The palisade (cortical) cells and the medulla stain dark violet. The oil droplets stains dark in color. (Fig. 2.1, 2.2)
Turbinaria ornata
The plant body is solid cylinder and appears lobed in transactional view. The epidermis is thin with small squarish cells. The cortical zone comprises fairly large, angular, thin wall compact cells. Medulla is wide and includes small, slightly thick walled compact cells.
Nile Blue Stain
Nile Blue stains cortex and medulla dark purple. The cell wall appears thick. Within the medullary cells are seen some granular or crystalline bodies. This is the area where oil droplets accumulates during stressful condition. (Fig. 3.1, 3.2)
Toludine Blue O
In Toludine blue staining, the epidermis appears dark. The cortex and medulla appears bright purple. The cells inclusions are not visible (Fig. 4.1, 2).
Photo graphs show esterification (biodiesel and residues layer), biodiesel production Fig 5.(a): Transesterification (b): Algal biodiesel Biodiesel can be produced from macro algae because of lipid contents. Sijama stated that docosahexaenoic acid (DHA) was a polyunsaturated fatty acid composed of 22 carbon atoms and six double bonds that belonged to the so called ω-3 group. They also reported that fish oil was the major source of DHA, but alternatively it might be produced by using of organisms. Marine organisms might contain large quantities of DHA and were considered a potential source of this important fatty acid. Some of these organisms could be grown heterotrophically on organic substrates without light. It has been reported that macro algae contain lipid content of 1.3-7.8% (dw). In addition in heterotrophic condition lipid content can be more in algae. The samples harvested are expected to produce lipid fractions containing Omega-3 fatty acids. After extraction and esterification to form the methyl esters, gas chromatographic analyses show (Fig 6 and 7) that the ω-3 fatty acids may constitute as much as higher of the total fatty acid fraction. They are generally contained in phospholipids, glycolipids, mono, or triglycerides and sulfolipids, or as the free acids, but are not limited to these forms (Long, T.V.,) The composition and structure of fatty acid esters, such as unsaturation degree and carbon.
chain length, determine the properties (e.g., cetane number, viscosity, cold flow, oxidative stability, and iodine value) of biodiesel.We therefore analysed the fatty acids of individual lipid classes from both Turbinaria and Sargassum. The fatty acid profiles of two Macro algae are presented (Table 2 and 3) respectively. However, saturated fatty acids (SFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA) displayed a significant interactive effect of growth modes.
The overall distribution of total fatty acid (TFA) in the lipid classes of both macro algae is depicted (Table 4). It was observed that Turbinaria ornata could produce large amount of poly unsaturated fatty acids (PUFA) as the major storage lipid which accounted 80.44% than the Sargassum ilicifolium is about 75.98% respectively. Since intracellular lipids can be enhanced by controlling the conditions such as nutrient limitation, limitation of light and salt stress, it can be expected that the TAG yield of macro algae can further be increased by the same approaches.
Fourier Transformer InfraRed (FTIR) gives five maxima of the generation of volatile products can be found in the spectrum. All collected extractions give bands at 674 and 2957 cm-1 (Fig ) so all are cis isomer, as expected from algal lipid because trans isomers have a strong nearby 970 cm-1 band and a weak nearby 3012 cm-1 while cis isomers gave medium nearby 720 and 3012 cm-1.bands An analysis of the IR spectrum obtained (Fig. 8 and 9) for the main composition stage, reveals the existence of the absorption bands characteristic of these five different bonds: • C?O: The main characteristic of the IR spectra of carbonylic compounds (aldehydes, acids, etc.) is the strong C?O stretching absorption band in the region of 1870-1540 cm-1. In the case of esters, this band appears in the 1750–1735 cm-1 • C-O-C: corresponding to ethers. These stretching vibrations produce a strong band in the 1200–900 cm-1 region. • C-H: absorption bands characteristic of the vibrations of C-H bonds, as an example, 2960 and 2875 cm-1 correspond to the asymmetric and symmetric vibrational modes of methyl groups, respectively, and 2929 and 2850 cm-1 correspond to the asymmetric and symmetric vibrational modes of methylene groups, respectively. • CO2: they produce strong bands in between 2800-2000 cm-1 as well as in 700 cm-1 region.
•H2O: the adsorption bands of water can be observed in the range of 1800-1200cm-1
CONCLUSION
Algae are an economical choice for biodiesel production, because of its availability and low cost. In this paper we have presented the preliminary results of an ongoing study on biodiesel produced from two macroalgae Sargassum ilicifolium and Turbinaria ornata. Many researchers reported that microalgae might better for higher biodiesl production. But research has not done yet in this regard. Till date there is no adequate research work or publications in India regarding macroalgal based biofuel Production. Our experimental results recorded maximum biodiesel production from macroalgae that confirms the previous findings. This may be a First report in India that the biodiesel producing macroalgae, their detailed ultrastructure and the lipid droplets storage. If we go further to demonstrate this model in the coastal India, there will be a larger opportunities to produce cost effective biofuel or biodiesel and glycerine production as well as the rural employment creation.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareVALUE ADDED PRODUCTS OF TOMATO AND ITS QUALITY CHARACTERISTICS
English7783V.Saradha RamadasEnglish T.ThilagavathiEnglishIn India, nearly 7.1 million tones of tomato are produced annually from an area of 5.4 lakh hectares. Tomato known as poor man‘s apple are cultivated on 70000 acres of land in the state with an average yield to 140000 tones of tomatoes a day being produced. Excess tomato production leaves Tamil Nadu farmers disappointed (www.thehindu.com). Tomato is a good source of vitamins, minerals and other useful substances like fiber which are necessary for good health. Tomato contains lycopene and betacarotene pigment. Lycopene is one of the most important components in tomato as it has been found to have a vital role in protecting humans from various types of cancer including colorectal, prostate, breast, endometrial and lung. Value addition to tomato through processing can be brought about by converting them into value added products. Sun Drying is found to be one of the ways to provide antioxidant product at low price to the community. Hence, tomato was dried and developed the various products using dried tomato and dried peel.
EnglishTomato, Cultivation, Lycopene, DryingINTRODUCTION
The top five leading fruit-producing countries are the United States, China, Turkey, Italy and India. Tomato production has been observed in 144 countries. The total global area under tomato is to the tune of 46.16 lakh hectares and the global production is to the tune of 1279.93 lakh tones. The major country China had a fruit production of 30,102,040 mt in 1,255,100 hectares. India accounts for only 8.6mt while China, the top producer, recorded an output of 33.7mt (FAOSTAT Database, 2004). In India, nearly 7.1 million tones of tomato are produced annually from an area of 5.4 lakh hectares. Tomato occupies second position amongst the vegetable crops in terms of production. On an average, about 10,800 tones of tomatoes are exported annually from India. Tamil Nadu occupies seventh position in production of tomato among the state. Tomato occupies around 0.25 lakh hectares in the state with an average yield of 12, 500kg per hectare and an annual production of annual three lakh tones. Tomato known as poor man‘s apple are cultivated on 70000 acres of land in the state with an average yield to 140000 tones of tomatoes a day being produced. Excess tomato production leaves Tamil Nadu farmers disappointed (www.thehindu.com). Tomato is a fruit used as a vegetable mainly because of the low sugar content. It is consumed both in fresh as well as processed forms. Tomato varieties are now available with double the vitamin-C, 40 times more vitamin-A, high levels of anthocyanins and 2-4 times more lycopene compared to that in traditional varieties. Tomato contains over 80 nutrients beneficial to human (Potty, 2009) Tomato is a good source of vitamins, minerals and other useful substances like fiber which are necessary for good health. Tomato contains lycopene and beta-carotene pigment. Lycopene is one of the most important components in tomato as it has been found to have a vital role in protecting humans from various types of cancer including colorectal, prostate, breast, endometrial and lung. Being a powerful antioxidant it has the ability to protect living cells and other structures in the body from oxygen damage and maintain DNA integrity in white blood cells. Lycopene is also believed to be able to activate cancer preventing phase 11 enzymes. There is another view, which gives credit to other components present in tomato along with lycopene for the anticancer activity of the fruit. Potty (2009) emphasized that lycopene is also linked to improve skin health by virtue of its ability to protect against undesirable UV ray exposure. (www.indiaagronet.com) Value addition to tomato through processing can be brought about by converting them into various products like tomato ribbon chips, rice and sago tomato vadagam and sweet tomato. The simplest and inexpensive method of processing tomato is drying. This can be achieved by processing the tomato by simple techniques and try to improve the shelf-life of the tomatoes. Thus the tomato products can be made available even in the off-season. Keeping these in mind the present study is focused with the following objectives. To
Develop an economically viable product to manage tomato wastage and
Observe the shelf-life and analyze the selected nutritional quality of the developed products.
METHODOLOGY
Sun-drying is an ancient method used in Indian households to dry fruits and vegetables. The removal of moisture from any substance is called dehydration. If the moisture in fruits and vegetables is reduced to a certain level by drying method, the shelf life can be increased. It is found to be one of the ways to provide antioxidant product at low price to the community. Hence, tomato was dried and developed the various products using dried tomato and dried peel. The steps involved in drying of tomato and list of products developed are shown in Plate I.
1. Drying of tomato: The variety of tomato selected for the powdering was country tomato (Namdhaari) which is available in the selected area. Tomato was washed swell, deseeded and dried in the sunlight. The dried tomatoes were powdered. Tomato products were developed and standardized using the dried tomato powder. Products like sago tomato vadagam, rice tomato vadagam and tomato ribbon chips were developed using the tomato powder.
2. Drying of peel: In order to make use of the peel the tomato was washed, blanched and peeled out the skin. The peel was sun dried and used for the product development of kulkanth. The tomato pulp, after removing the peel was used for the preparation of tomato sauce and pickle. Preservative was added to improve the shelf life of the tomatoes. Vinegar is used to develop a product. The concentration of vinegar in finished product should not be less than two per cent in order to ensure good result.
A) SENSORY EVALUATION OF DEVELOPED PRODUCTS
Sensory evaluation of all the developed products was done by a group of 20 selected panel members using a score card with the criteria of appearance, flavor, texture, color and taste. The panel members evaluated each sample on a specific scale for a particular characteristic indicating the rating of the samples. In order to identify the best quality product the mean acceptability scores were calculated.
B) ANALYSIS OF NUTRITIVE VALUE OF THE DEVELOPED PRODUCT
Since tomato is a good source of vitamin C and the pigment lycopene the analysis of these nutrients was done with the help of Post Harvest Technology Centre, Tamil Nadu Agricultural University, and Coimbatore. The major nutritive content of lycopene and vitamin C were analyzed in raw tomato, dried tomato powder and the products developed using dried tomato powder and the dried tomato peel. Dye method was followed for analyzing lycopene and vitamin C in fresh as well as processed tomato product and procedure followed for the analysis are given in Appendix III.
C) OBSERVATION OF SHELF LIFE OF THE DRIED TOMATO
All the developed products were stored in polyethylene cover and in air-tight plastic container for 60 days at room temperature. Quality of the stored products in terms of organoleptic quality and moisture content were assessed for every 15 days till two months. Hot air oven method was followed for finding out the moisture content in dried tomato powder. In order to ensure the quality of the dried tomato powder and product developed by using dried tomato powder and tomato peel, observation was made to assess whether the product is free from insect, fungal in mold growth over a period of 60 days.
RESULTS OF THE STUDY
1. Yield of tomato powder and peel
Yield of tomato powder and peel is shown in Table I. The study revealed that a yield of six and seven per cent of tomato powder was obtained respectively from one kg of country and hybrid tomato and there was not much difference between the two varieties. The yield of tomato peel was found to be 21 per cent and 29 per cent respectively in country and hybrid tomato since the quantity of moisture content in peel was very much less than the tomato pulp. The yield of tomato powder from one kg of country tomato was 45.5g whereas hybrid tomato was 54.5g. The yield of tomato peel from one kg of country tomato was 9g whereas hybrid tomato was 17g. Aeration sun-drying (ASD) was tried for the preparation of tomato powder. From one kg of fresh tomato 50-55g of dehydrated powder was obtained with desirable quality. The overall acceptability score was found to be 8.3 on 9- point hedonic scale. The cost of drying was Rs.1.23 per kg (Gupta et al., 2006).
2. Nutritive value of developed product
Table II predicts the lycopene and vitamin C value of the developed product using the dry tomato powder. Raw tomato contains 2.42 and 2.04mg of lycopene and vitamin C respectively whereas tomato powder had 18.55 and 14.80mg of lycopene and vitamin C respectively in 100g due to loss of moisture content. The Polychemical Information Center states that cooked tomato based foods have greater health benefits, compared to uncooked, because the heating process makes all carotenoids (including lycopene) more easily absorbed by the body (http://recipes.suite101.com). The tomato powder had 14.80mg of vitamin C per 100g whereas the dry vadagam containing 30g of tomato made with rice and sago had 3.63 and 5.38mg of vitamin C per 100g respectively. The vitamin C content of dry and fried tomato vadagam made with rice and sago was found to be less than the dry tomato powder. However, frying also further degraded the value of vitamin C content. This is also proved by the study of Murator et al. (2008). He stated that temperature was directly related to ascorbic acid loss.
3. Sensory evaluation of developed product
Table III shows the sensory evaluation of developed products. Sensory evaluation is done for all products which scored, 4.9, 4.7, 4.6, 4.5 and 4.1, tomato powder, sago tomato vadagam, rice tomato vadagam, tomato ribbon chips and kulkanth. Tomato powder scored high value among the developed products. This organoleptic score indicates that the dry powder and the product developed using tomato powder were highly acceptable as the total mean score was ranged from 20.6 to 24.7 out of 25 scores. Tomato ketchup was prepared using seven different recipes and they are explored for organoleptic evaluation to select the best recipe along with recovery and processing cost. It was found to be best in terms of recovery cost and organoleptic scores (Charusinghal et al., 2005).
4. Shelf life of dry tomato powder
Table IV reveals that the quality of dry tomato powder in packed in polyethylene cover and airtight plastic container for two months. Shelf life of the tomato powder was assessed by sensory evaluation and found out its quality over a period of 60 days. Shelf life of the dry tomato powder prepared by using dried tomatoes was assessed over a period of two months. Quality of the packet products were checked for every 15 days till two months. The moisture content of the tomato powder not changed up to one month but after the end of 45 days, one per cent of moisture has been increased. The maximum of mean score of five was awarded for the appearance, color, texture, flavor and taste for the dry tomato powder over a period of 60 days by the panel members. This indicates that the powder stored in air tight plastic containers and polyethylene cover showed no difference indicating the shelf life of the products is at acceptable standard. There was no insect, fungal or mold growth till 60 days. The study can be extended further to evaluate how long the tomato powder is safe without any spoilage. There was not much difference between the materials air tight plastic container and polyethylene pack used for storing tomato dry powder and in both the storage it was safe over a period of two months.
DISCUSSION
It is noted that the yield of tomato powder from one kg of country tomato was 45.5g whereas hybrid tomato was 53.5kg. The yield of tomato peel from one kg of country tomato was nine grams whereas hybrid tomato was 17g. The study indicated that six to seven per cent of dried tomato powder was the yield from both varieties, 21 per cent of dried tomato peel was the yield from the country tomato, whereas 29 per cent of yield was derived from hybrid tomato. Raw tomato contains 2.42 and 2.04mg of lycopene and vitamin C per 100mg. The lycopene and vitamin C content in 100mg of tomato powder was 18.55mg and 14.80mg respectively. The difference between raw and dried tomato vadagam may be due to loss of moisture content. Rice tomato vadagam dry contains 0.84mg and 1.75mg of lycopene and vitamin C respectively per 100g whereas 2.03mg and 3.63mg of lycopene and vitamin C in 100g of fried vadagam. Sago tomato vadagam contains 0.31mg and 3.03mg per 100g in dry whereas 2.11mg and 5.38mg of lycopene and vitamin C in 100g of fried vadagams. Tomato ribbon chips contain 0.48 and 5.92 mg of lycopene and vitamin C per 100g. Kulkanth made with tomato peel contains 0.90 and 6.63 mg lycopene and vitamin C per 100g. The mean total score for the products sago tomato powder, rice tomato powder and tomato ribbon chips had a score of 23 and found to be highly acceptable. The moisture content of the tomato powder not changed upto one month but after the end of 45 days, one per cent of moisture has been increased. The shelf life study revealed that when dried tomato powder is stored in polyethylene cover and air-tight plastic container for one month, they scored 25. Though after 30 days, there is slight difference in flavor and taste and scored 23 and the tomato powder, the products developed were acceptable at the high level.
CONCLUSION
Thus, the study will promote employment opportunities to rural farmers, reduce tomato wastage, develop new value added product and available during off-season and raise the standard of living of rural population.
ACKNOWLEDGEMENT
The investigator extends her heartful thanks to Post Harvest Technology Centre, Tamil Nadu Agricultural University, Coimbatore for providing all amenities needed for the successful conduct of the study.
Englishhttp://ijcrr.com/abstract.php?article_id=2162http://ijcrr.com/article_html.php?did=21621. Gupta, S., Ghuman, S.S. and Sandhu, K.S. Preparation of tomato powder on small scale, Journal of Food Science and Technology (2006), 43(1), 31-33.
2. Charusinghal, Singh P. and Singh, J.P. A study on techno-economic analysis of recipes for processing of tomato ketchup, Beverage and Food World (2005), 32(5), 24-25.
3. Muratore, G., Nobile, M.A.D., Buonocore, G.G., Lanza, C.M. and Asmundo, C.N. The influence of using biodegradable packaging fibres on the quality decay kinetic of plum tomato, Journal of Food Engineering (2005), 67(4), 393-399.
4. Muratore, G., Rizzo, V., Licciardello, F. and Maccarone, E. Partial dehydration of cherry tomato at different temperature and nutritional quality of the products, Food Chemistry, (2008), 111(4), 887-891.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareEFFICACY OF EXTERNAL FOCUS OF ATTENTION IN IMPROVING BED MOBILITY IN HEMIPLEGIC PATIENTS.
English8491Baburaja Panicker Harilal MPTEnglish Vijesh P.V MPT M Phil.English Sasidharan Tapan MPTEnglish Sankar Sahayaraj MPTEnglishObjectives: To find out the efficacy of external focus of attention in improving bed mobility in
hemiplegic patients.
Method: A sample of 30 patients satisfying the inclusion criteria were randomly assigned into a control
group and an experimental group of 15 patients each. The control group received bed mobility training
without external focus of attention. In experimental group external focus of attention was used to give bed
mobility training . The treatment was for 6 days. The outcome measure taken was Trunk Control Test. Pre
and post values were taken on the first day and sixth day of treatment.
Results: Statistical analysis using Wilcoxon signed-ranks test and Mann-Whitney ?U‘ test showed
significant difference in improvement in bed mobility status of patients treated with external focus of
attention.
Conclusion: External focus of attention for training bed mobility seems to be beneficial for hemiplegic
patients.
EnglishExternal focus of attention, Hemiplegic patient, Bed mobility.INTRODUCTION
Stroke (Cerebrovascular accident - CVA) is defined as -A focal (or at times global) neurological impairment of sudden onset, and lasting more than 24 hours (or leading to death), and of presumed vascular origin1 Several population-based surveys on stroke were conducted from different parts of India. During the last decade, the age adjusted prevalence rate of stroke was between 250-350/100,000. The ratio of cerebral infarct to hemorrhage was 2.21. The estimation of stroke mortality derived was 102000 deaths, which represented 1.2% of total deaths in the country2 Although stroke rehabilitation has to some extent progressed over the years, there is still lack of enthusiasm in pursuing the goal of ensuring that each patient recovers his best possible function3 Often however, a gradual return of some motor abilities occurs in weeks and months after injury. The severity of trunk impairment is usually less than more distal musculature. Poor recovery of trunk muscle performance results in a severe disability and a reduction in the activities of daily living. In stroke rehabilitation, trunk muscle performance is an important factor in predicting the functional outcome 4 .
In patients with hemiplegia trunk control is an indispensable basic motor ability for execution of many functional tasks5 . Turning to the intact side and sitting up over the side of the bed are critical to the re establishment of independence. However for individuals who cannot perform these without assistance, it is important to help them into the sitting position. In this way training to re-establish swallowing, communication, visual scanning of the environment, attentional capacity and the ability to balance and move can start early6 . Motor learning research with its focus on discovering laws and principles underlying the acquisition of motor skills had little impact on clinical applications in physical therapy7 . Feedback is an effective method in motor learning which can be used in the rehabilitation of neurological patients.8-11. Feedback can be extrinsic or intrinsic. During early motor learning the therapist will provide extrinsic feedback to shape performance. During early intervention visual inputs are critical for motor learning12 Learning strategies may be used for the patients with hemiplegia who is required to transfer across a variety of surfaces including obstacles of varying heights and distances apart. An external focus of attention may yield a learning advantage for the activity being practiced because the automatic control processes underlying the activities such as those associated with balance and stability are allowed to operate freely13. Careful planning of content, scheduling and attentional focus induced by the feedback can enhance the effectiveness of training considerably. However, research examining these issues in stroke patients is scarce14 . In this study bed mobility training is done with external focus of attention. For external focus the subject was asked to concentrate on a point made by a laser pointer, which reflects the performance of the patient. So the incorporation of attentional focus strategy in bed mobility training is studied here.
METHODOLGY
Study was approved by the institutional ethical committee, written informed consent was obtained from the participants. Study was conducted in the physiotherapy departments of General Hospital and Ananthapuri Hospital and research institute Trivandrum. Study design was experimental and 30 stroke patients with hemiplegia due to MCA territory involvement were selected using non probability convenient sampling and then they were assigned to control and experimental group of 15 each using random assignment. The selection criteria include male patients with first episode of stroke within the age group of 45 to 55 years. They should come under Brunnstrom‘s recovery stage15 2 and should have the ability to roll to the affected side and should have a MMSE score16 minimum 20. Subjects having Hemi neglect, visual agnosia, visual field defects, aphasia, cerebral atrophy, cerebellar lesions, history of seizures, pressure sores cardio respiratory complications and orthopedic complications were excluded from the study. Outcome measure used was Trunk control test (TCT). The inter-rater reliability and validity of TCT have already been reported17,18 TCT examines four axial movements; rolling from supine position to weak side (T1) and to strong side (T2), sitting up from a lying down position (T3) and sitting in a balanced position on the edge of the bed with feet off the ground for 30 seconds (T4). The TCT score is the sum of the scores obtained on the four tests (range, 0-100). Material used was a laser pointer having a diode with wavelength 630 to 680 nm, and with a maximum output of < 1mw. Laser pointer was modified for attaching to the subjects body Before starting the procedure subjects were tested using TCT. The control group received bed mobility training which includes rolling to the weak side and then to strong side, getting up from supine and sitting balance exercise. In addition to this subjects received exercises which include ROM, bridging and positioning strategies. For experimental group bed mobility training was done with external focus of attention. For this the laser pointer was strapped to the subject‘s chest and markings were made on the ceiling as well as on both side walls. When the laser is put on subject has to concentrate on the laser point on the ceiling and should try to move the point to the marking on the side wall. For training sitting balance laser is projected to the front wall where three markings were made. For all the exercises subject is given assistance for completion of task. When he gains control of the movement, the assistance is gradually lowered. The procedure was repeated for six days.
RESULTS
The results were analyzed with SPSS version 16 for windows using Wilcoxon signed-ranks test and Mann-Whitney ?U‘ test. The mean pre-test score of TCT for the control group was 34 (SD±18.33) and for the post-test was 59.07(SD±16.68). These values for the experimental group were 31.53(SD 14.55) and 84.67(SD 11.84) respectively. From the analysis it can be seen that there is statistically significant difference between the pre and post test values of control group as well as experimental group. Furthermore when analyzing post-test values of the control and experimental group by Mann-Whitney test, statistically significant difference is there. This indicates that external focus of attention was effective than internal focus of attention in bed mobility training for the subjects. From the item wise analysis it can be seen that for the control group and for the experimental group all the items were improved significantly from the pretest value. When analyzing the post-test values of the control and experimental group it is clear that there is significant difference between the post-test values of control and experimental group in all items except in T1. (Rolling to weak side from supine) .It means that external focus of attention was better over conventional treatment in improving all the components of TCT except T1.
DISCUSSION
Numerous studies have shown that instructions that direct individuals‘ attention to the effects of their movements on the environment thereby inducing a so called external focus of attention, were more effective for learning than instructions that direct attention to the movement themselves or inducing an internal focus.19-23 After stroke, intrinsic feedback system may be compromised making it difficult for the person to determine what needs to be done to improve performance. Extrinsic feedback may thus be even more important to people with stroke.14 According to constrained action hypothesis24,25 an internal attentional focus, or focus directed to something close to the body results in participants subtly interfering in relatively automatic control processes. As a result of this interference the degrees of freedom of the motor system are presumably constrained in such a way that the rate and effectiveness of the system to regulate movement or maintain balance is subtly compromised. Presumably there is a delicate balance between the conscious process and automatic process, which can be interfered with or overridden when the participant consciously intervene in the control process. This type of interference seems to occur to a lesser extent when the participants‘ attention is directed further away from the body and to the external effects. Common coding theory26,27 provides a possible explanation for the advantages of focusing on the effects of one‘s own movements, rather than on the movement themselves. Because according to this theory, perception and action requires a common representational medium, efferent and afferent codes are stored in the form of distal events. From this point of view, it makes sense to assume that action will be more effective if they are planned in terms of their intended outcome or effect, rather than in terms of the specific movement patterns. When participants were practiced rolling, getting up from lying down and balance in sitting position, the internal focus used by the participants constrained the motor system by interfering with natural control processes, whereas the laser marker as the effect of the movement (external focus) allowed the automatic control processes to regulate the movements. Furthermore the subjects who used laser markings, the actions are planned in terms of there intended outcome, that is to the point to which the marker should move, rather than subjects who concentrated on their movement patterns. On further analyzing the results it can be seen that all components of TCT improved significantly between post-test scores of control and experimental group except T1 (Rolling towards weak side from supine). This may be probably because due to the selection criteria of subjects. All subjects included in the study were with a minimum score 12 for T1. With 6 days of treatment most of the subjects in control and experimental group improved to the highest score, which are 25. So there was not a significant difference between the post-test scores of control and experimental group for the component T1 One of the limitation in this study was it calculated only a short term outcome of the treatment. Effects of this improvement on the late functional outcome in subjects were not studied. Also sample size was small and subjects included were limited to males with MCA territory infarction. So studies can be conducted in large sample size in other areas of rehabilitation like hand rehabilitation, gait training using external focus strategy. Relationship between early independency in bed and late functional outcome using this strategy also can be studied.
CONCLUSION
Bed mobility training using external focus was better than the training was it is not used. When the subjects concentrated on the external effects it provided a better learning advantage for the subjects. External focus of attention for training bed mobility seems to be beneficial for hemiplegic patients.
ACKNOWLEDGEMENT
The authors are grateful to patients and staffs of General Hospital and Ananthapuri Hospital Thiruvananthapuram, Management and staff, Sumandeep Vidyapeeth, Mr. Asok School of Behavioral Sciences, MG University and Mr. Shajith KP, Department of Telemedicine, Medical College Aleppey for their support during the study.
Englishhttp://ijcrr.com/abstract.php?article_id=2163http://ijcrr.com/article_html.php?did=21631. World Health Organization. WHO steps Stroke Manual: The WHO Stepwise approach to stroke surveillance. WHO 2006; p. 5
2. Anand K, Chowdhary D, Singh KB, Pandav CS, Kapoor SK. Estimation of mortality and morbidity due to strokes in India. Neuroepidemiology 200; 20: 208-11.
3. Carr JH, Shepherd RB. A motor relearning programme for stroke, 2nd ed. London: Buttorworth Heinemann; 1987. p. 4-6.
4. Fujiwara T, Sonoda S, Okajima Y, Chino N. The relationships between trunk function and the findings of transcranial magnetic stimulation among patients with stroke. J Rehabil Med 2001; 33: 249-55
5. Wade DT, Langton R. Motor loss and swallowing difficulty after stroke: frequency, recovery and prognosis. Acta Neurol Scand 1987; 76: 50-54.
6. Carr JH, Shepeard RB. Neurological rehabilitation: optimizing motor performance, 1st ed. India: Buttorworth Heinemann 1998. p. 250.
7. Winstein CJ. Knowledge of results and motor learning: implications for physical therapy. Phys Ther 1991; 140-49.
8. Morris ME, Iansek R, Matyas TA, Summers JJ. Stride length regulation in Parkinson‘s disease. Brain 1996; 119: 551-68.
9. Cheng PT, Wang CM, Chung CY, Chen CL. Effects of visual feedback rhythmic weight shift training on hemiplegic stroke patients. Clin Rehab 2004; 18: 747-53.
10. Bourbonnias D, Bilodeau S, Lepage Y, Beadoin N, Gravel D, Forget R. Effects of force feedback treatments in patients with chronic motor deficits after stroke. Am J Phys Med Rehabil 2002; 81; 890-97.
11. Dursun E, Hamamci N, Donmez S, Tuzunalp O, Cakci A. Angular biofeedback device for sitting balance of stroke patients. Stroke 1996; 27: 1354-57.
12. O‘Sullivan SB, Schmitz TJ. Physical rehabilitation: assessment and treatment, 4th edition. India: Jaypee; 2001. p. 514-45.
13. McNevin NH, Wulf G, Shea CH. Increasing the distance of an external focus of attention enhances learning. Psycholog Res 2003; 67: 22-29.
14. Van Vliet PM, Wulf G. Extrinsic feedback for motor learning after stroke: What is the evidence? Disab Rehab 2006; 28: 831-40.
15. Sawner KA, La Vigne JM. Brunnstrom‘s movement therapy in hemiplegia - a neuro physiological approach, 2nd ed. Philadelphia: Lippincott Williams and Wilkins; 1992. p. 41-42.
16. Folstein MF, Folstein SE, McHugh PR. "Mini-mental state". A practical method for grading the cognitive state of patients for the clinician. J Psychiatr Res 1975 Nov;12(3):189-98
17. Collin C, Wade D. Assessing motor impairment after stroke: A pilot reliability study. J Neurol Neurosurg Psychiatry 1996; 53: 576-79.
18. Franchignoni FP, Tesio L, Ricupero C, Martino MT. Trunk Control Test as an early predictor of stroke rehabilitation outcome. Stroke 1997; 28: 1382-85.
19. Wulf G, Shea CH, Park JH. Attention and motor performance: Preference for and advantages of an external focus. Res Q Exerc Sport 2001; 72: 335-44.
20. Tang QP, Yang QO, Wu YH, Wang GQ, Huang ZL, Liu ZJ, Huang XS, Zhou L, Yang PM, Fan ZY. Effects of problem oriented willed-movement therapy on motor abilities for people with post stroke cognitive deficits. Phys Ther 2005; 85: 1020-33.
21. Fasoli SE, Trombly CA, Ticle-Degned L, Verfaellie MH. Effects of functional reach in persons with and without cerebrovascular accident. Am J Occup Ther 2002; 56: 380- 90.
22. Zachary T, Mercer J, Bezoidis N. Increased movement accuracy and reduced EMG activity as the result of adopting an external focus of attention. Brain Res Bull 2005; 67(4): 304-9.
23. Wulf G, Weight M, Poulter D, McNevin N. Atentional focus on suprapostural tasks affects balance learning. Quart J Experim Psychol 2003; 56: 1191-211.
24. Wulf G, Prinz W. Directing attention to movement effects enhances learning: A review. Psychomet Bull Rev 2001; 8: 648- 60.
25. Wulf G, McNevin N, Shea CH. The automaticity of complex motor skill learning as attentional focus. Quart J Experim Psychol 2001; 54: 1143- 54
26. Prinz W. A common coding approach to perception and action. In: Neumann O, Prinz W, editors. Relationships between perception and action. Berlin: SpringerVerlag; 1990. p. 167-201.
27. Prinz W. Perception and action planning. Eur J Cogn Psychol 1997; 9: 129-54.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareTHE NEED OF PHARMACOVIGILANCE OF DISULFIRAM - A REVIEW
English9297Prafulla ManeEnglish Ambrish ThosarEnglish Kavita AtreEnglish Rahul MayeeEnglishA good pharmacovigilance practice is the need of time to ensure that the drugs in the market are fulfilling the purpose for what they are meant. Similarly, the literature survey highlights the miracle drug i.e. disulfiram which is showing promising results and giving a sigh of relief to the families from the rural areas of our country by enabling the alcohol dependent patients to quit alcohol. . Disulfiram is a drug used to support the treatment of chronic alcoholism by producing an acute sensitivity to alcohol. Disulfiram is also being studied as a treatment for cocaine dependence, as it prevents the breakdown of dopamine. Several studies have reported that it has anti-protozoal activity as well. There is no tolerance to disulfiram: the longer it is taken, the stronger its effects. The adverse drug reactions (ADRs) of disulfiram are themselves used as a treatment for alcoholism. So, the more we get pharmacovigilant about the drug the more it will be fruitful for us to treat the patient. Hence, we can call disulfiram as one of the life saving drugs for the alcohol addicts knowing the consequences of excessive consumption of alcohol.
EnglishPharmacovigilance, disulfiram, alcoholism, adverse drug reaction.INTRODUCTION
Pharmacovigilance is the science and activities relating to detection, assessment, understanding and prevention of adverse effects or any other possible medicine related problem. It aims in making best use of medicines for the treatment or prevention of diseases. Though it is not intended to harm the patients during medicine use process, unfortunately some medicines often cause problems. Significant harm to even a few patients not only destroys the credibility of the medicine but the patients too, loose trust on the care professional or the health system.[1] It is the process of identifying and responding to the issues of drug safety through effects usually adverse.[5] Need for Pharmacovigilance Systems To be eternally vigilant to ensure that medicines, which are developed for treatment of diseases, actually do not do more harm than good, is one of the important pre-requisites for the progress of medicine. Already, at least in the U.S.A., figures as high as 3 to 5% of hospital admissions have been attributed to iatrogenic diseases, that is., those caused by drugs. The science and systems used for systematically identifying and correlating drugs and side-effects and taking corrective actions fall under the discipline of Pharmacovigilance.[2] Key objectives pharmacovigilance can be briefly defined as the process of evaluating and improving the safety of medicines.[11]
Pharmacovigilance Process
Finding the risks of drugs
Medicines have helped to bring improved health and longer life to human beings. Medicines affect the lives of hundreds of millions of people every day. But they are not without risk, and have caused, do cause and will continue to cause harm to many people. There are also large numbers of people who experience no evident effect at all from the drugs they take.[3, 4] Pharmaceutical companies are required by law in all countries to perform clinical trials, testing new drugs on people before they are made generally available. The manufacturers or their agents usually select a representative sample of patients for whom the drug is designed — at most a few thousand — along with a comparable control group. The control group may receive a placebo and/or another drug that is already marketed for the disease.[3, 4] Clinical trials do, in general, tell us a good deal about how well a drug works and what potential harm it may cause. They provide information which should be reliable for larger populations with the same characteristics as the trial group - age, gender, state of health, ethnic origin, and so on.[3, 4] Good Pharmacovigilance will identify the risks and the risk factors in the shortest possible time so that the harm can be avoided or minimized. The integration of Pharmacovigilance may be crucial to the success of public health programmes involving medicines. Under the stimulus and coordination of World Health Organization (WHO) and its Collaborating Centre for International Drug Monitoring (Uppsala Monitoring Centre), National programmes are built up to make it a true international necessity.[1]
The specific aims of pharmacovigilance are to:
Improve patient care and safety in relation to the use of medicines and all medical and paramedical interventions. Improve public health and safety in relation to the use of medicines; contribute to the assessment of benefit, harm, effectiveness and risk of medicines, encouraging their safe, rational and more effective (including cost-effective) use. Promote understanding, education and clinical training in pharmacovigilance and its effective communication to the public.[6]
Pharmacovigilance is mainly governed by:
The pharmaceutical industry (prime responsibility), competent (regulatory) authorities, WHO collaborating Centre for International Drug Monitoring (in Uppsala), Council for International Organization of Medical Sciences - CIOMS Working Groups on drug safety plays an advisory role in harmonization of pharmacovigilance practices.[1]
Pharmacovigilance in India
Pharmacovigilance is fastest emerging as an important approach for the early detection of unwanted effects of the drugs and to take appropriate regulatory actions if necessary .This may ensure the safer use of drugs.[7] Historically, Indian market has always, except in very few cases, seen the launch of only products, which have been earlier approved and marketed in U.S.A., Western Europe or Japan. Until now, the time lag between the first marketing of a new drug in a foreign country and India has been on an average around 4 years, and hardly any new drug was introduced for the first time in India. In that kind of scenario, it was not too critical that there was in place a system of pharmacovigilance in India, since reports of side-effects from outside India would have helped our regulatory agencies to assess the rationale of continuing the drug in the Indian market. Thus in the past, action on marketed drugs has been triggered on the basis of reports on the harmful effects of drugs marketed abroad. In a few cases, drugs, which have been banned or withdrawn in foreign markets, were allowed to be kept in the market in India. For example, Chloramphenicol, Phenyl Butazone, Clioquinol, Phenformin, Cisapride, all continue to be prescribed in India on the basis of a conscious decision by the Regulatory Agency that the benefit to risk ratio is in favor of the former.[8] The evolution of a new Patent regime in the Indian Pharmaceutical Industry (the Post-2005 scenario) as a consequence of India being a founder member of WTO, and her obligations under Trade Related Intellectual Property Rights and Services (TRIPS), makes it incumbent that India can no longer copy patented products and market them without license from the innovator company. The leading Indian companies realizing the compulsions of the new regime have already initiated investments of substantial resources for the discovery and development of new drugs needed for both Indian and International markets. This in turn means that during the coming years Rand D by the Indian Pharmaceutical companies will hopefully lead to new drugs based on pre-clinical and clinical data generated mostly in India. In such cases, the Indian regulatory agencies cannot count on the experience of other markets to assess the incidence and prevalence of adverse reactions from drug usage, and therein lays the importance of a properly designed pharmacovigilance system in India. For an effective Pharmacovigilance system to be functional and efficient all the stakeholders need to be alert and attentive throughout the lifetime of the drug in the market.[8]
Need for ADR reporting
The most frequent question that arises in pharmacovigilance is the need to monitor the adverse reactions of the drugs, even though their safety profiles have already been studied adequately before their commercial release. The answer to this question is to make the drugs safer. In addition, the formal therapeutic trials are conducted in carefully controlled conditions; in highly selected and limited number of patients, so that the exact safety profile of the drug in the real life situations is not known. Children, pregnant women, and elderly are not included in clinical trials for ethical reasons. Therefore, the safety of the drug in these cases remains unknown until its release. Another important drawback of clinical trials is that they can only report adverse reactions that appear within the finite duration of trial. Delayed reactions would be missed. Reporting of adverse drug reactions is done by mainly two methods - spontaneous and intensive. Though plagued by numerous problems like low yield of reports, sub-optimal quality and imperfect nature, these have often served to be a useful source of data or provided early warning signals for the drug related regulatory actions.[9] At the time a drug is approved knowledge about its risk is incomplete. Tests in animals are necessary and useful to discover toxic effects, but do not allow sufficient conclusions about human safety. Clinical studies focus on demonstrating efficacy statistically instead of comparing benefits and ADRs with those of existing drugs. The small number of patients involved in, and unsatisfactory length of, clinical studies limit the value of their findings. Thus, pre-approval clinical data include only information about the most common ADRs. In addition, specific doses are used and patients who may be at greater risk from ADRs are usually not studied during the development of a drug, e.g. young children, elderly people, pregnant or lactating women, patients concomitantly using other drugs or other therapies, patients with complicated disease conditions, sub-populations carrying known and relevant genetic polymorphism and patients of different racial and/or ethnic origins.[10] Thus, clinical studies give very limited information about risk and efficacy in real life conditions. Reporting of harm related data from clinical studies needs improvement.[11] Here arises the research question that, why Pharmacovigilance of Disulfiram? Now the answer to this is very simple as we move into the details of the drug. Disulfiram is a drug used to support the treatment of chronic alcoholism by producing an acute sensitivity to alcohol. Trade names for disulfiram in different countries are antabuse and antabuse manufactured by Odyssey Pharmaceuticals. Disulfiram is also being studied as a treatment for cocaine dependence, as it prevents the breakdown of dopamine (a neurotransmitter whose release is stimulated by cocaine); the excess dopamine results in increased anxiety, higher blood pressure, restlessness and other unpleasant symptoms. Several studies have reported that it has anti-protozoal activity as well.[12]
Pharmacology:
Mechanism of action:
Disulfiram inhibits aldehyde dehydrogenase, the oxidative enzyme of acetaldehyde, a metabolite of alcohol. The latter is accumulated in the blood, thus producing unpleasant symptoms of disulfiramalcohol reaction when a patient has taken small amounts of alcohol. Absorption: Rapidly absorbed from the GIT; peak plasma concentrations after 8-10 hrs (oral). Metabolism: Reduction to diethyldithiocarbamate by the glutathione reductase system in the erythrocytes.
Excretion:
Urine (as metabolites); exhaled gas (carbon disulphide).
Dosage:
Disulfiram is supplied in 200 mg, 250 mg, and 500 mg tablets. The usual initial dose is 500 mg for 1 to 2 weeks, followed by a maintenance dose of 250 mg (range 125 mg– 500 mg) per day. The total daily dosage should not exceed 500 mg.[13]
How does Disulfiram work?
Under normal metabolism, alcohol is broken down in the liver by the enzyme alcohol dehydrogenase to acetaldehyde, which is then converted by the enzyme acetaldehyde dehydrogenase to the harmless acetic acid. Disulfiram blocks this reaction at the intermediate stage by blocking the enzyme acetaldehyde dehydrogenase. After alcohol intake under the influence of disulfiram, the concentration of acetaldehyde in the blood may be 5 to 10 times higher than that found during metabolism of the same amount of alcohol alone. As acetaldehyde is one of the major causes of the symptoms of a "hangover" this produces immediate and severe negative reaction to alcohol intake. Some 5–10 minutes after alcohol intake, the patient may experience the effects of a severe hangover for a period of 30 minutes up to several hours. Symptoms include flushing of the skin, accelerated heart rate, shortness of breath, nausea, vomiting, throbbing headache, visual disturbance, mental confusion, postural fainting, and circulatory collapse. Disulfiram should not be taken if alcohol has been consumed in the last 12 hours. There is no tolerance to disulfiram: the longer it is taken, the stronger its effects. As disulfiram is absorbed slowly through the digestive tract and eliminated slowly by the body the effects may last for up to two weeks after the initial intake; consequently, medical ethics dictate that patients must be fully informed about the disulfiramalcohol reaction.[14]
Adverse Drug Reactions:
The most common adverse drug reactions are drowsiness, headache, tiredness, fatigue, metallic or garlic taste in the mouth, vision changes, decreased sexual ability, mental or mood changes, trouble in breathing, increased heart rate, muscle weakness, numbness or tingling of hands and legs and several others. Whereas the moderately common occurring ADRs are severe dizziness, poor memory, dark urine, bad body odor. Also there are some rarely occurring ADRs such as yellowing of eyes, skin and nails, which are the symptoms of jaundice, severe allergic reactions such as skin rash, itching, swelling specially on the face, tongue or throat.[15] There are also certain special instructions to be followed while consuming disulfiram. Do not drink any alcoholic beverages while taking disulfiram, during the 12-hours period before you take your first dose, and for several weeks after stopping the drug. Always carry an identification card stating that you are taking disulfiram and indicating the doctor to be contacted in an emergency. Disulfiram makes some patients drowsy. Do not drive a car or operate dangerous machinery until you know how this drug affects you. Do not stop taking disulfiram without consulting your doctor. Special dietary instructions should be followed. Avoid sauces, vinegars, and all foods and beverages containing alcohol. If you remember a missed dose within 12 hours of when you should have taken it, take the missed dose. However, if it is more than 12 hours since you should have taken it, omit the missed dose and take only the next dose at its regularly scheduled time. Do not take more than one dose in a 12-hour period.[16]
CONCLUSION
The survey of literature showed that there is a need to be pharmacovigilant about disulfiram and such other drugs used for the treatment of alcoholism. As we now know that the adverse drug reactions of disulfiram is the key for the treatment of the alcoholics there must be more action taken for the ADR reporting of this miracle drug. The percentage of population quitting alcohol due to the disulfiram treatment is much greater than the other remedies. So, if more attention is focused on the pharmacovigilance of the drug it may be possible to report more such ADRs which might be beneficial for treatment of alcohol dependent patients.
Englishhttp://ijcrr.com/abstract.php?article_id=2164http://ijcrr.com/article_html.php?did=21641. The Importance of Pharmacovigilance, Safety Monitoring of Medicinal Products. WHO 2002, the Uppsala Monitoring Centre, WHO Collaborating Centre for International Drug Monitoring.
2. The Importance of Pharmacovigilance, World Health Organization 2002
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5. Tsintis P. and Mache E. CIOMS and ICH Initiatives in Pharmacovigilance and Risk Management Overview and Implications European Agency for the Evaluation of Medicinal Products. United Kingdom Drug Safety 2004; 27 (8):509-517.
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13. Kuruvilla A. Pharmacovigilance - Review Topic. Rational Drugs; Issue 26; Oct - Dec. 2006.
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15. Johnsen J., Stowell A., Bache -Wiig JE., Stensrud T., Ripel A. and Morland J. A double-blind placebo controlled study of male alcoholics given a subcutaneous disulfiram implantation. Br J. Addict 1987; 82:607-613.
16. Fuller RK. and Roth HP. Disulfiram for the treatment of alcoholism: An evaluation in 128 men. Ann Intern Med 1979; 90:901- 904.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareIN-VITRO ANTIBACTERIAL SCREENING OF THE EXTRACTS OF SWERTIA CHIRAYITA LINN. AGAINST MRSA (METHICILLIN RESISTANT Staphylococcus aureus)
English98104Abdul LatifEnglish Sumbul RehmanEnglishShamim AhmadEnglish Asad U KhanEnglishAqueous and ethanolic extract of Chirayita (Swertia chirayita Linn.) were screened for their antibacteria activity Methicillin Resistant Staphylococcus aureus (MRSA). Kirby Bauer‘s Disk Diffusion method an Broth Serial Dilution method according to CLSI Guidelines by W.H.O were used. It was compared with the Standard drug-Vancomycin and Plane control-DMSO (the solvent used). The Resistant drug– Methicillin was also used to confirm the resistance of MRSA strains used. The prepared plates were incubated and results were evaluated by measuring the Zone of Inhibition - ZOI (in mm.) of drug extract. MIC and MBC was also carried out against the resistant strain. All the experiments were conducted in triplicates and in sterilized conditions. The results were analyzed statistically using gpaid software and it was found that MRSA strain was sensitive to S.chirayita showing a significant ZOI as compared to the standard drug. Present investigation indicates that S.chirayita possesses antimicrobial properties and hence can be used for future natural plant based antimicrobial agents. Thus the study provides an in-vitro proof of its antibacterial activity against MRSA. However further investigations are needed to find out its pharmacological details and to make it fit for use, for the welfare of humankind. So, that it can be used
against the infectious diseases caused by the resistant strain of Staphylococcus aureus safely and effectively.
EnglishSwertia chirayita Linn. MRSA, Methicillin, VancomycinINTRODUCTION
Over the last three decades Methicillin Resistant Staphylococcus aureus is responsible for causing several difficult-to-treat infections in humans. Also known as Multidrug resistant S.aureus or Oxacillin resistant S.aureus (ORSA) emerged as a noscomial pathogen in early 1960s [1] it is resistant to a large group of antibiotics called beta-lactams, which include the penicillin and the cephalosporins. A study led by the CDC (Centre for Disease Control) and published in the October 17, 2007 issue of the Journal of the American Medical Association estimated that MRSA would have been responsible for 94,360 serious infections and associated with 18,650 hospital stay-related deaths in the United States in 2005. These figures suggest that MRSA infections are responsible for more deaths in the U.S. each year than AIDS. In 2007, the CDC reported that MRSA causes 19,000 deaths every year in the US, which is more than HIV/AIDS cases [2]. The studies done so far about the increasing incidence of MRSA infection are motivating the researchers to find out new pharmacological components of ?natural origin‘. Antibiotics provide the main basis for the therapy of microbial (bacterial and fungal) infections. Since the discovery of these antibiotics and their uses as chemotherapeutic agents there was a belief in the medical fraternity that this would lead to the eventual eradication of infectious diseases. However, overuse of antibiotics has become the major factor for the emergence and dissemination of multi-drug resistant strains of several groups of microorganisms [3]. The worldwide emergence of Resistant strains of Staphylococcus aureus has become a major therapeutic problem. The increasing failure of chemotherapeutics and antibiotic resistance exhibited by pathogenic microbial infectious agents has led to the screening of several medicinal plants for their potential antimicrobial activity [4]. Researchers are increasingly turning their attention to herbal products, looking for new leads to develop better drugs against multidrug resistant microbe strains. A wide group of medicinal plant preparations are available that have been used over the centuries almost exclusively on the basis of empirical evidence [6]. The primary benefits of using plant-derived medicines are that they are relatively safer than synthetic alternatives, offering profound therapeutic benefits and more affordable treatment [7]. The World Health Organization estimates that 65%-80% of the world's population use traditional medicine as their primary form of health care. The use of herbal medicine, the dominant form of medical treatment in developing countries, has been increasing in developed countries in recent years [8]. Swertia chirayita (Family Gentianaceae) is used in traditional medicines since the period of Discorides. It is rich in a wide variety of secondary metabolites such as alkaloids and flavonoids, which have been found in vitro to have antimicrobial properties [6]. Phytochemical analysis reveals that it contains a yellow bitter acid Ophelic acid (C15H 20O 13), two bitter glucosides-chiratin (C26H 48O 15) and amarogentin (C32H 38O 16); gentiopicrin, two yellow crystalline phenols, xanthone-swerchirin [9-13]. It has been found to have anti-carcinogenic, antiviral, antihelminthic, antimalarial, antipyretic, anti-inflammatory, anti-arthritic activity [14-20]. There are several reports on the antimicrobial activity of its extracts [21]. These evidences contribute to support and quantify the importance of screening S. chirayita. The pharmacological potential of Swertia chirayita reveals that its antibacterial activity towards a large number of bacterial strains, but towards MRSA its activity has not been done so far, so this was an attempt to screen its in-vitro antibacterial activity against the resistant bacteria.
MATERIAL AND METHODS
The herb was procured from the local market Baradari of Aligarh city and was properly identified by the Botanical literature available and then confirmed by Prof. S. H. Afaq from the Pharmacognosy section, Department of Ilmul Advia, Aligarh Muslim University, Aligarh. Voucher specimens (SC-0100/09-G) were preserved in the herbarium of Medicinal Plant Lab in the Department of Ilmul Advia, F/O Unani Medicine, Aligarh Muslim University, Aligarh for future reference.
Preparation of plant extract
Two different extract were prepared for analysis in the present study viz. aqueous extract and ethanolic extract as per W.H.O guidelines with some minor modifications [22].
For aqueous extract:
10 gm of the powdered drug and 150 ml of the Double Distilled Water (DDW) were put into a soxhlet apparatus. The solvent was boiled at 400C and refluxed for a period of 150 min (eleven extraction cycles).The extract was filtered and evaporated to dryness under reduced pressure in the Lyophilizer (Macro Scientific works, Delhi). It was redissolved in DMSO (Dimethyl Sulphoxide) to the desired concentration (20 mg/ml) for the study.
Similarly for the ethanolic extract: 10 gm of the powdered drug and 150 ml of the ethanol (Solvent) were put into a soxhlet apparatus and the same procedure was repeated as stated above.
Microorganisms used
Clinical strains of MRSA isolated from various sources viz. pus (Pus culture: PC), urine (Urine Culture: UC) and control strains N315, Mu50, ORSA (Oxacillin resistant Staphylococcus aureus) of the tested microorganisms were obtained from Department of Microbiology, JNMC and Hospital, AMU; Microbiology Section, Institute of Ophthalmology, JNMC and Hospital, AMU, Aligarh. The bacterial cultures were grown in Nutrient Broth (M002 Himedia Labs, Mumbai, India) and incubated at 370C for 24 hours, followed by frequent sub culturing to fresh media and were used as test bacteria. The bacterial cultures were checked to confirm the presence of sufficient number of bacterial cells on nutrient broth and maintained on nutrient agar slant.
Antimicrobial activity
Antimicrobial assay of the crude extracts was performed against pathogenic strains by Kirby Bauer‘s Disk diffusion method and Agar well method [23, 24]. The nutrient agar plates were swabbed with a suspension (106 cfu/ ml) of the bacterial strains. The wells of the equivalent size were prepared with the help of a cork borer and the drug (40µl) was poured in the respective well with the help of a micropipette. Finally, the resistant antibiotic–Methicillin disks (SD137, Himedia Labs, Mumbai, India) were placed on the prepared plates with sterile forceps and pressed properly to make complete contact with the surface of the medium. Later on these plates were kept at room temperature for 30 minutes (Pre-diffusion time) and then incubated at 370C for 24 hours. The antibiotic disks (6 mm) were used as Positive Control while the solvent used for diluting the test drug was used as the Negative Control. The diameters of the inhibition zone – Zone of Inhibition (ZOI) in mm was measured and is given in Table-1. The experiment was done in triplicate and the mean values were calculated.
Determination of minimum inhibitory concentration (MIC)
Minimum inhibitory concentration of various extracts against tested microorganisms was determined by broth dilution method [25]. 96- well microtitre plates were used, 50 µl of standardized suspension of a strain (106 cfu/ml) (cfu-colonies forming unit) was added to each tube containing extracts at various concentrations. The plates were incubated at 37oC for 24h and observed for visible growth. The minimum inhibitory concentration (MIC) is taken as the lowest concentration of the extracts at which there is turbidity after incubation, the values are given in Table-2. Minimum Bactericidal Concentration (MBC) was further determined from the same isolates MBC is the minimal concentration of drug needed to kill most (99.9%) of the viable organisms after incubation for 24 hours.
STATISTICAL ANALYSIS OF DATA
All the values have been expressed as Mean ± SEM (Standard error of mean). Statistical significance was determined by one way ANOVA (Analysis of variance) using g-paid software for calculation.
RESULT
The aqueous and ethanolic extracts of S.chirayita exhibit varying degree of inhibitory effect against all tested pathogenic strains (Table-1). The MIC of crude extracts of the drug was determined at the concentrations ranging from 19.53 to 5000 µgm/ml (Table-2). The aqueous extract produced zone of Inhibition in the range of 8.8 to 12.0 mm for all the tested strains while ethanolic extract produced comparably larger zone against them in the range of 23 to 30 mm, the results were found to be significant with p-value>0.0001. The minimum inhibitory concentration varies in the range of 625 to 1250 µgm/ml for the aqueous extract while it was lesser towards ethanolic extract in the range of 78.12 to 312.5 µgm/ml of the S.chirayita extract. Aqueous extract of the test drug sample was found to have lesser activity as compared to ethanolic extract, but both extract showed larger zone as compared to the inhibitory zone of Vancomycin, which is considered as effective drug for MRSA at present. DMSO- the solvent used to dissolve the drug was also tested; it has not shown any inhibitory effect towards the tested bacteria. This confirms that the inhibitory activity of the extract was only due to the drug used and not due to the solvent in which it was dissolved. Further the resistance to Methicillin was maintained on the nutrient plate, as the antibiotic disk does not produce any inhibitory effect, this confirms the resistance of the tested strains.
DISCUSSION
Medicinal plants are naturally gifted with invaluable bioactive compounds which form the backbone of traditional medicines. Many infectious diseases have been known to be treated with herbal remedies throughout the history of mankind [26]. In the present investigation, in vitro antimicrobial efficacy of the crude extracts of S.chirayita was quantitatively assessed on the basis of inhibition zone and minimum inhibitory concentration. The study shows that the crude ethanolic extract of S.chirayita showed more pronounced antimicrobial activity as compared to aqueous extracts, and when efficacy of either extract was compared to Standard drug ?Vancomycin‘ it was found to be quite efficacious showing a greater ZOI. The result of the present investigation suggests that S.chirayita is a potent natural source of biologically active compounds from herbal medicines, which may potentially prove to be efficient natural antimicrobial agents. This antibacterial property of S.chirayita can be attributed to the presence of the biologically active constituents present in it like amarogentin, swerchirin, triterpenoids, xanthones, ophelic acid, gentiopicrin [27]. This study also supports the previous antibacterial activity of the chirayita extract towards gram positive bacterial strains [28] and verifies the claims of traditional medicine for their use in various infectious diseases [29-31]. However exactly which constituent is helping in killing these dreadful bacteria and in which manner, this part of the study is further need to be explored. And more investigations are needed in this direction before its use in clinics, as the present study is just an in-vitro proof about its antibacterial efficacy; many studies regarding its clinical use are still needed to be done.
CONCLUSION
With the emergence and widespread occurrence of multi drug resistant bacteria focus has now been shifted in exploring natural compounds that may combat drug resistance problem. Clearly strategic planning for search on fundamental and safe integration of efficacious medicine into conventional medical practice is needed. On the other hand the drugs used in traditional system of medicine could be systematically explored for novel bioactive compounds which can inhibit drug resistance bacteria or which can enhance activity of antibiotics. Such compounds or herbal medicine may find application in combination with antibiotic for the treatment of bacterial diseases. The study suggests that S.chirayita exhibit antimicrobial properties against MRSA which is an emerging cause of a number of infectious diseases and has developed resistance to the synthetic antibiotics. The potential antimicrobial activity of S.chirayita towards the infectious micro-organism explains the basis for its use in future in combating the disease caused by such dreadful bacteria.
Englishhttp://ijcrr.com/abstract.php?article_id=2165http://ijcrr.com/article_html.php?did=21651. Tyagi A, Kapil A, Singh P: Incidence of Methicillin Resistant Stahylococcus aureus (MRSA) in Pus Samples at a Tertiary Care Hospital: AIIMS: New Delhi: JIACM: 2008; 9 (1): 33 - 35
2. en.wikipedia.orgi date of access : 12-12- 2010.
3. Khan R, Islam B, Akram M, Shakil S, Ahmad A, Ali SM, Siddiqui M and Khan AU: Antimicrobial Activity of Five Herbal Extracts Against MultiDrug Resistant (MDR) Strains of Bacteria and Fungus of Clinical Origin: Molecules 2009;14: 586- 597
4. Chahal K. J., Sarin R., and Manvimalwal: Efficacy of clerodendrum inerme l. (garden quinine) against some human pathogenic strains: International Journal of Pharma and Bio Sciences 2010; 1(4): 219 - 223
5. Cos P, Vlietinck AJ, Berghe DV, Maes L: Anti-infective potential of natural products: How to develop a stronger in vitro ?proofof-concept‘: Journal of Ethnopharmacology 2006; 106: 290 - 302
6. Bandow JE, Brotz H, Leichert LIO, Labischinski H and Hecker M: Proteomic approach to understanding antibiotic action: Amicro. Agents. Chemother. 2003; 47: 948 - 955
7. Kumar VR, Kumar S, Shashidhara, Anitha : Modulation of CNS Activity By Traditional Medicines: A Review On Neuro Pharmacognosy: International Journal of Current Research and Review2011; 3 (1):39-48 8. Lewis K, Ausubel FM : Prospects of plant derived antibacterials: Nat. Biotechnol. 2006; 24: 1504-1507
9. Anonymous: The Wealth of India: A dictionary of Indian Raw materials and Industrial Products: CSIR: New Delhi1976; X: 77-81
10. Bhattacharjee SK and De LC: Medicinal Herbs and Flowers: Aavishkar Publishers: Jaipur 2005; 236- 237
11. Chopra RN, Chopra JC, Handa KL and Kapur LD: Indigenous drugs of India: U N Dhur and Sons Pvt. Ltd: Calcutta 1958: 292- 293
12. Nadkarni KM: The Indian Materia Medica: Bombay Prakashans Pvt. Ltd 2000; I: 1184
13. Wallis TE: Textbook of Pharmacognosy: CBS Publishers: Delhi 1985: 317-319
14. Saha, Prosenjit MS, Das A, Das PC and Das S: Evaluation of anticarcinogenous activity of Swertia chirayita-Buch.-Ham., an Indian Medicinal Plant, on DMBA-induced mouse skin carcinogenesis model: Phytotherapy Research.2004; 18 (5):373-378
15. Verma H, Patil PR, Kolhapure RM, Gopalkrishna V : Antiviral activity of the Indian medicinal plant extract, Swertia chirata against herpes simplex viruses: A study by in-vitro and molecular approach: Indian Journal of Medical Microbiology 2008;26 (4): 322-326
16. Zafar I, Lateef M, Khan MN, Jabbar A and Akhtar MS: Antihelminthic activity of Swertia chirata against gastrointestinal nematodes of sheep: Fitoterapia 2006; 77 (6): 463-465
17. Panda, Ashok K, Tripathy PC and Nandy A : Clinical evaluation of Swertia chirata in the treatment of P.vivax malaria: Aryavaidyan 2005;19 (1): 39-42
18. Bhargava S, Rao SP, Bhargava P, Shukla S: Antipyretic potential of Swertia chirata Buch Ham. Root extract: Scientia Pharmaceutica 2009; 77: 617-623\
19. Kumar M.L.R.S., Yadav A.K., Saxena A. and Paul B.N. : Modulation of Interleukin- 1β, Interleukin-4, Interleukin-6, Interleukin- 10, Tumor Necrosis Factor-α and Interferon- γ by aqueous extracts of Swertia chirayita: Indian Journal of Pharmacology 2002; 34: 141-155
20. Kumar MLRS, Paul BN, Asthana R, Saxena A, Mehrotra S and Rajan G : Swertia chirayita Mediated Modulation of Interleukin-1β Interleukin-6, Interleukin-10, Interferon-γ, and Tumour Necrosis Factor-α in Arthritic Mice: Immunopharmacology and Immunotoxicolog 2003; 25 (4): 573-583
21. Bonjar S: Evaluation of Antibacterial Properties of Some Medicinal Plants Used in Iran. :J.Ethnopharmacol 2004; 94: 301-305
22. Anonymous: Quality control methods for medicinal plant materials: Word Health Organization: Geneva 1998: 25-28
23. National Committee for Clinical Laboratory Standards: Methods for Disk Susceptibility Tests for Bacteria that Grow Aerobically: NCCLS Document M2-A7: National Committee for Clinical Laboratory Standards: Wayne, USA: 2000; 7
24. Ananthanarayan R and Paniker‘s: Textbook of Microbiology:8th edition:Publishers University Press: Hyderabad 2009: 618
25. Jennifer MA: Determination of Minimum Inhibitory Concentrations: Journal of Antimicrobial Chemotherapy 2001; 48, (SI): 5 -16
26. Chahal KJ, Sarin R, and Manvimalwal: Efficacy of clerodendrum inerme l. (garden quinine) against some human pathogenic strains: International Journal of Pharma and Bio Sciences 2010; 1(4): 219 - 223
27. Evans WC and Trease: Pharmacognosy: 15th edition: Elsevier publications: Delhi 2009: 417
28. Alam KD, Ali MS, Parvin S, Mahjabeen S, Akbar MA and Ahmed R: In vitro antimicrobial activities of different fractions of Swertia chirata ethanolic extract: Pak. J. Biol. Sci. 2009; 12: 1334-1337
29. Ghani HN: Khazainul advia: Idara Kitabul Shifa: New Delhi 1921; II: 595-596
30. Momin KM: Tohfatul Mominin (Persian) Matba Hasni 1272: 75
31. Nadkarni KM: The Indian Materia Medica: Bombay Prakashans Pvt. Ltd. 2000; I: 1184
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30HealthcareNUTRIGENOMICS: THE EMERGING FACE OF NUTRITION
English105115Sheweta BarakEnglish Deepak MudgilEnglish BS KhatkarEnglishNutrigenomics is the science that examines the response of individuals to dietary compounds, foods and diets using post-genomic and related technologies. It aims to determine the influence of common dietary ingredients on the genome. This new understanding of health and disease will lead to innovative developments in human medicine, agricultural biotechnology, food quality and safety, new food products, biomarker development and nutritional interventions. Unbalanced nutrient intakes are known to be associated with the development of chronic diseases demonstrating that dietary chemicals have direct effects on molecular genetic processes. Nutrigenomics is a potentially promising development in emerging functional food and nutraceutical research.
Englishnutrigenomics, nutrigenetics, diet, diseasesINTRODUCTION
Associations between diet and chronic disease have long been recognized through epidemiological studies. New genomic technologies have enabled to find out more about the basis of these associations through studies on the functional interactions of food with the genome at the molecular, cellular and systemic levels and, the ways in which individuals respond differently to different diets according to their individual genetic make-up. The completion of Human Genome Project, an effort to clone, map and sequence the entire human genome, has improved the understanding of nutrition. With the development of scientific and technological advances in the field of health and nutrition, much focus is now being directed towards the emerging field of Nutrigenomics which is being applied to agriculture (enhanced plants and animal food sources) and to human health1 . Nutrigenomics explores how nutrients or bioactive dietary compounds can influence gene expression (Fig 1) and an individual‘s risk to certain diseases. It has the potential to impact various parts of the food chain including the genetic modification of crops and animal feeds, the development of nutrigenomic tests to monitor food safety and personalization of diets, as well as the identification of nutritional supplements and nutrigenomic based products which may offer potential health benefits. This new understanding of health and disease will lead to innovative developments in human medicine, agricultural biotechnology, food quality and safety, new food products, biomarker development and nutritional interventions. The long-term aim of nutrigenomics is to understand how the whole body responds to real foods using an integrated approach termed ??systems biology‘‘2. Over the past few decades, epidemiological and clinical studies have indicated many relations between nutrition and health. Links have been established between dietary habits and degenerative diseases like cardiovascular diseases; type 2 diabetes and cancer3 . Unbalanced nutrient intakes are known to be associated with the development of chronic diseases demonstrating that dietary chemicals have direct effects on molecular genetic processes. From a Nutrigenomics perspective, food is enormously complex. Nutrigenomics looks beyond one gene at a time because most of the major health conditions involve tens and hundreds of genes.
Nutrigenomics and other „omics technologies
Nutrigenomics is the science that examines the response of individuals to dietary compounds, foods and diets using post genomic and related technologies, i.e. genomics, transcriptomics, proteomics and metabolomics. It aims to determine the influence of common dietary ingredients on the genome, and attempts to relate the resulting different phenotypes to differences in the cellular and/or genetic response of the biological system4 . More practically, nutrigenomics describes the use of functional genomic tools to probe a biological system following a nutritional stimulus that will permit an increased understanding of how nutritional molecules affect metabolic pathways and homeostatic control. Well-known examples of nutrigenomics research are the analyses of the huge array of gene polymorphisms relating to obesity and diabetes, the genetic polymorphism of enzymes for nutrient metabolism (e.g., enzymes for folate metabolism), and the genes involved in sodium sensitivity in hypertension5 .
Nutrigenetics examines the effect of genetic variation on the interaction between diet and disease or on nutrient requirements. It embodies the science of identifying and characterizing gene variants associated with differential responses to nutrients, and relating this variation to disease states6 . Genetics has a pivotal role in determining an individual‘s risk of developing a certain disease7 . The transcriptome is the complete set of RNA that can be produced from the genome.
Transcriptomics is a very valuable way of beginning to understand how nutritional exposure influences gene expression on a genomic scale. It is possible to group genes of interest for particular metabolic processes and capture information from all of these at once to see how the cell is functioning at any given time or under certain conditions. Such techniques are aided by the commercial development of chips orientated around particular metabolic or functional systems.
Proteomics is the study of the proteome, and it addresses three categories of biological interest: protein expression, structure and function8 . It attempts to characterize all proteins in a biological sample, including their relative abundance, distribution, posttranslational modifications, functions, and interactions with other biological molecules. Dietary components can also modify the translation of RNA to proteins and the posttranslational events, which can affect protein activity9 .The presence or absence of certain key proteins can give information about the early stages of disease. Currently, the most widely used technologies for proteomics are two dimensional (2D) gel electrophoresis to separate the proteins in a complex mixture isolated from cells or tissues, and specialized mass spectrometry techniques as protein identification tools 10,11 . One of the newest ?omics? technologies in nutrition is metabolomics. It focuses on the analysis of metabolites, the metabolome. It tries to measure the level of all substances (other than DNA, RNA or protein) present in a sample; the metabolome comprises the complete set of metabolites synthesized by a biological system12 . Metabolomics examines global patterns of metabolites present in the cell or in body fluids in response to specific dietary exposures. Metabolomics is a useful tool for generating individual metabolite profiles, such as complete plasma lipid (ie, cholesterol, triglycerides) and vitamin profiles.
Epigenetics is the study of modifications to the genome which are copied from one cell generation to the next but which do not involve changes to the primary sequence. These changes, mediated through modification of chromatin proteins such as histones and through the methylation of DNA, contribute to the regulation of transcription and provide a way for the genome to ?learn from experience?, regulating gene expression in response to dietary and other exposures and leading to altered cellular phenotypes associated, for example, with chronic disease or ageing. All of these ?- omics tools? have been used to study in detail the molecular responses to food substances or the early stages of disease in common dietrelated conditions.
System biology The long term aim of nutrigenomics is to understand how the whole body responds to real foods using an integrated approach termed 'systems biology'. Systems biology is an approach for studying biological systems that analyzes multiple macromolecular species (DNA polymorphisms, RNA, protein, metabolites, etc) in one experiment. It is a holistic approach to study biological systems.
NUTRIENT-GENE INTERACTION
Genes are turned on and off according to metabolic signals that the nucleus receives from internal factors, e.g. hormones, and external factors, e.g. nutrients, which are among the most influential of environmental stimuli. Genomes evolve in response to many types of environmental stimuli, including nutrition. Therefore, the expression of genetic information can be highly dependent on, and regulated by, nutrients, micronutrients, and phytochemicals found in food13. Unbalanced diets alter nutrient gene interactions, thereby increasing the risk of developing chronic diseases. Rare genetic differences are called mutations, and common genetic variations (occurring in more than 1% of a population) are called polymorphisms. One major initiative is cataloguing the simplest form of these variations (called single-nucleotide polymorphisms, or SNPs). A SNP (pronounced 'snip') occurs when only a single nucleotide (chemical letter) in the DNA sequence varies. There are thought to be some 100,000 to 300,000 SNPs in human genes, which may either influence phenotype directly or be used as markers by researchers when they look for important genetic variants14 . Some SNPs directly alter a metabolic response to a nutrient, rather than changing the requirement for it. MicroRNA (miRNA) expression may be influenced by dietary manipulation15, but little data are available describing miRNA-level modulation of genes of metabolism. Finally, genetic variation influences eating behaviors16,17 but these effects have not been systematically explored. Other types of genetic variation include copy-number polymorphisms (CNPs).Numerous dietary components can alter genetic events, and thereby influence health. Dietary chemicals can affect gene expression directly or indirectly18,19. At the cellular level, nutrients may: 1) act directly as ligands for transcription factor receptors; 2) be metabolized by primary or secondary metabolic pathways, thereby altering concentrations of substrates or intermediates involved in gene regulation or cell signaling; or 3) alter signal transduction pathways and signaling. From the molecular standpoint, nutrients are considered to be signaling molecules that, through appropriate cellular sensing mechanisms, result in translation of these dietary signals into changes in gene, protein, and metabolite expression20,21 . Methylene tetrahydrofolate reductase (MTHFR) enzyme catalyzes the reaction that produces 5- methyl tetrahydrofolate. MTHFR has a role in supplying 5methylenetetrahydrofolate, which is necessary for the remethylation of homocysteine to form methionine. Methionine is essential to many metabolic pathways including production of neurotransmitters and regulation of gene expression. Folate is essential to the efficient functioning of MTHFR. For the MTHFR gene tow important SNPs has been well recognized: C677T (cytosine to thymidine substitution resulting in the conversion of an alanine to valine) and A1298C (adenine to- cytosine substitution resulting in the conversion of an alanine to glutamic acid)22. The presence of C677T or A1298C mutations is associated with reduction in MTHFR enzyme activity and impairs folate accumulation, which may cause increases homocysteine concentration in plasma, a risk factor for venous thromboembolic and ischemic arterial diseases23. The beneficial effect of folic acid supplementation (1 mg/day for 3 months) on plasma homocysteine level has been shown in a geno type stratified, randomized, double-blind, placebo controlled trial24. MTHFR is also involved in maintenance genomic CpG methylation patterns and prevention of DNA strand breaks, these mutations are associated with increased risk of neural tube defects and some types of cancer25 . Manganese super oxide dismutase (MnSOD) is a mitochondrial enzyme that plays a key role in detoxification of reactive oxygen species. A polymorphism valine to alanine substitution in this enzyme alters its transport into mitochondria, which has been associated with increased risk of breast cancer9 . Enzymes that utilize and metabolize vitamin B12 have been associated with NTDs, increased risk of Down syndrome and colon cancer. For example, a common polymorphism in the HFE gene (Cys282Tyr) is associated with iron storage disease hereditary haemochromatosis, leading to an iron accumulation in the liver, heart and endocrine glands. This protein is an important regulator of cellular iron homeostasis and has role in intestinal iron absorption by regulating the interaction of the transferrin receptor with transferrin25 . Cytochrome P450s (CYPs) enzymes play a central role in the oxidative biotransformation of steroids, prostaglandins, nutrients, drugs, chemicals and carcinogens. Several dietary factors can alter the expression of CYP isoforms. CYP1A2 plays an essential role in the metabolism of wide range of drug and chemical substances. For example, CYP1A2 activates dietary carcinogens such as aromatic amines, but also detoxifies compounds such as caffeine. Low-activity CYP1A2 genotype with an increased risk of myocardial infarction suggests that this enzyme detoxify a substance, which may be an important risk factor in the population. Indeed, individuals with a lowactivity CYP1A2 genotype are at a greater risk of coffee- associated heart disease. As caffeine is the main substance in coffee and is detoxified by CYP1A2, it may be an important risk factor for heart disease in certain population26 . Glutathione S transferase (GST) enzyme is a superfamily of enzymes that play an important role in the detoxification of several dietary compounds. GSTM1, GSTT1 and GSTP1 are isoforms of this enzyme. The GSTM1 and GSTT1 null genotype have been associated with both an increased and a decreased risk of some types of cancers such as breast cancer26,27. Some components such as dietary isothiocyanates that are found in cruciferous vegetables are eliminated with GSTs enzymes.
Peroxisome proliferator-activated receptors (PPARs) are nuclear receptor supper family that plays an essential role in fatty acid oxidation, glucose, and extracellular lipid metabolism. PPARs are the best-known fatty-acid-regulated nuclear receptors. One of the three members of the PPARs family regulates many genes involved in fatty acid metabolism. PPR-α (PPARA) plays a central role in lipid oxidation and inflammation, whereas PPAR-γ is involved in adipocytes differentiation, glucose and lipid storage, and inflammation. PPAR-δ (also known as PPAR-β), may has a crucial role in development, lipid metabolism, and inflammation. These receptors bind to fatty acid and regulate the expression of genes involved in fatty acid transport and metabolism. PPARs family also involve in activation of about 300 genes28. The PPAR-α gene has a polymorphism at codon 162 (Lue162Val) that has been associated with changes in total cholesterol, LDL-associated cholesterol, and Apo B concentrations. The less common V162 allele is associated with significantly higher serum concentration of total cholesterol, LDL cholesterol, Apo B, and Apo C-III than in carriers of L162 allele, especially in men. For individuals with the common L162 allele, increased intake of polyunsaturated fatty acids (PUFAs) had little effect on fasting triacylglycerol concentrations. In those with the less common V162 allele, however, fasting triacylglycerol concentrations fell abundantly with increasing PUFA intake29 . Moreover, dietary chemicals can directly affect signal transduction pathways. For example, green tea contains the polyphenol, 11- epigallocatechin-3- gallate (EGCG) that EGCG inhibits tyrosine phosphorylation of Her-2/neu receptor and epidermal growth factor receptor that reduces signaling via the phosphatidyl inositol 3-kinase (PI- 3)-AKt kinase-NF-kB pathway. Activation of the NF-kB pathway is associated with some types of breast cancer30,31,32 . Approximately 40 micronutrients are needed in human diet and each has specific role to play in maintaining genomic stability (Table 1).
NUTRIGENETICS AND DIET RELATED DISEASES
Nutrigenetics and Galactosemia, PKU and Favism
Different mutations in galactose-1-phosphate uridyltransferase (GALT) gene35 , 36 , phenylalanine hydroxylase gene37, and Glucose- 6-phosphate dehydrogenize (G6PD) gene 38 , 39,40 resulted in Galactosemia, Phenylketonuria (PKU), and Favism diseases, respectively. Other examples of enzymes polymorphisms include Lactase-phlorizin hydrolase gene (LPH) polymorphisms that show how SNPs alter gene expression. This polymorphism is in the upstream of the lactase-phlorizin hydrolase gene (LPH) associated with hypolactasia and changes tolerance to dietary lactose (milk sugar, LPH hydrolyzes lactose into glucose and galactose) and allows different expression of the LPH41 .
Nutrigenetics and Cardiovascular diseases
Cardiovascular diseases are a group of multifactorial conditions associated with atherosclerosis, thrombosis and hypertension. The link between genetic variations in the APOE gene and heart disease has been found to be significant. Apolipoprotein E (ApoE) gene has three different alleles (ε2, ε3, ε4). Persons with ε4 variant respond to a high-fat diet negatively with an increased risk for coronary heart disease (CHD). In these individuals, low-fat diet should be useful23. Moreover, there is an important relationship between allelic variants in the ApoA1/C3/ A4/A5 genes and the effect of dietary fats on lipoprotein metabolism and CVD (cardio vascular diseases) risk. Linkage disequilibrium within Apo A1/C3/A4/A5 cluster has been represented to affect plasma lipid concentration and CVD risk. Apolipoprotein A- 1 is and is a key component of high-density lipoprotein particles (HDL). The locus of gene encoding APOA-1 is on chromosome 11q and highly polymorph and has a specific SNP in its promoter region37. An Adenin/Guanin substitution in the promoter region (-75bp) of the ApoA1 gene is common in different populations. The presence of A allele (A/A and A/G) has been associated with increased HDL cholesterol. Moreover, mild increase in APOA-1 concentrations in subjects with the G/G genotype was observed22,42. APOA-5 gene is also an important regulator of triglyceride (TG) - rich lipoprotein (TRL) metabolism27 .
Nutrigenetics and Diabetes II
Diabetes is a group of disorders that result in high blood sugar levels (hyperglycemia). The body either lacks the ability to make the hormone insulin or does not use it properly. Diabetes also increases the risk of other diseases such as heart disease, blindness, nerve damage and kidney damage. Type 2 diabetes develops over time as the body gradually stops producing enough insulin (abnormal 'insulin secretion') or the cells in the body stop using it properly (called 'insulin resistance'). A rare inherited form of type 2 diabetes, called Maturity-Onset Diabetes of the Young (MODY) can result from mutations in any one of at least six different genes43. In addition, mutations in genes in the mitochondria (inherited via the mother's egg) can cause another rare form of diabetes (accounting for about 1% of cases)44. To date, over 250 genes have been studied for their role in type 2 diabetes and the majority of studies have failed to find any association. A common genetic variation in the KCNJ11 gene appears to slightly increase risk45 , 46. The CAPN10 gene47 and the IRS-1 gene may also have a small effect but results are very inconsistent.
Nutrigenetics and Cancer
Cancer is a complex disease and how it develops and spreads is not fully understood. However, it is thought to be caused by damage to the DNA inside a person's cells, including mutations and other types of damage, which then cause some cells (cancer cells) to grow out of control. Most mutations are thought to arise during a person's lifetime (called 'somatic' mutations) but some people can be born with mutations ('germ line' mutations) that increase their risk of cancer, often at an unusually early age. Diet plays an important role in increasing or decreasing the risk of developing some cancers. However, there are significant uncertainties about the role of diet in cancer and the effects depend on the type of cancer. Mutations in genes which increase the risk of breast and ovarian cancer and of colorectal cancer are some of the best studied. Mutations in either of two genes called BRCA1 and BRCA2 have been associated with a lifetime risk of breast cancer of between 45% and 87%. Mutations in these genes are thought to account for about 5% of breast cancer cases, and also increase the risk of ovarian cancer. Also it has been reported that women who consume less fruits and vegetables than required reported to be at greater risk of developing breast cancer because of a polymorphism that causes a valine to alanine (Val to Ala) change in the ninth position in the signal sequence for the enzyme manganese dependent superoxide dismutase48 . Familial adenomatous polyposis (FAP) is a largely inherited form of colorectal cancer. About 0.5% to 1% of colorectal cancer is thought to be due to mutations in the BRCA1 and BRCA2 genes. Lynch Syndrome is a form of hereditary colorectal cancer (called 'hereditary nonpolyposis colorectal cancer' or HNPCC) associated with mutations in the family of MMR genes (including four genes: hMSH2, hMLH1, PMS1, PMS2). About 3-5% of colorectal cancer is thought to be due to Lynch Syndrome. Also variants of melanocortin1 receptor gene (MCIR) have been found to be associated with several types of skin and prostrate cancers49. Glutathione peroxide is a selenium-dependent enzyme that acts as an antioxidant enzyme. Polymorphism at codon 198 of human glutathione peroxides results in a substitution of proline to leucine amino acid, and has been associated with an increase risk of lung cancer. Investigators shown that persons with (Pro/Lue) genotype were at 80% greater risk for lung cancer and (Lue/Lue) genotypes were at 130% greater risk compared risk those with the (Pro/Pro) genotype.
Nutrigenetics and Obesity
Obesity has become a major public health problem. Mutations in the genes like leptin and leptin receptor genes have emerged as leading factors towards predicting obesity. In addition to this, mutations in melanocortin 4 receptor and melanocortin 5 receptor gene and in the noncoding regions of the gene for neuropeptide Y (NPYY5R) receptor has also shown to be strongly correlated with the risk of obesity50 .
PERSONALIZED DIET AS A HEALTH STRATEGY
In its simplest form, nutrigenomics is based on the idea that diet should be tailored to an individual's genetic make-up or genotype (this is sometimes called nutrigenetics). A person's genome is the inclusive set of all their 25,000 or so genes. The genes are the parts of the DNA sequence that contain the cell's instructions for making proteins. The study of the genome is called genomics. Nutrigenomics research may also include other biological measurements (not just a person's genetic make-up). In the future, some of these other measurements may also be used to 'personalize' nutrition or to help design new functional foods. To the food industry, nutrigenomics provides an opportunity to design new products, attempt new 'personalized' marketing strategies (based on genetic test results, or, in the longer term, on other biological measurements) and to claim that it is responding to public concern about the growing epidemic of diet-related disease. The aim is to prevent disease and improve quality of life through functional foods and tailored diets.
CONCLUSION
Nutrigenomics is a potentially promising development in emerging functional food and nutraceutical research. The ability of bioactive food components to influence gene expression patterns (nutrigenomics effects) is also a factor in determining the overall response. Finally, bioactive food components may influence protein synthesis, degradation, and posttranslational modification. Nutrigenomic studies involving calorie restriction, specific carbohydrate and fat features, and exercise interventions have uncovered mechanisms that up-regulate and down-regulate gene expression toward a beneficial state of health, especially in patients with metabolic syndrome, obesity, and type II diabetes. Understanding the interrelationships among human genetics diversity, genome function, and dietary components will enable precise manipulation of genome function and stability throughout the life cycle for optimal human health and disease prevention. Nutrigenetics is only beginning to claim its potential. One can visualize the development of beverages and foods either as preventive agents or for the treatment for individuals, families, or subgroups predisposed to a particular disease. Increased physical activity and recommended diets balanced in omega-3/omega-6 ratios can be the pillars for health promotion and prevention of multiple chronic diseases. As we advance our knowledge of gene-nutrient interactions, society will need to create or utilize appropriate social, ethical, legal, educational, and economic frameworks to gain the benefits of such knowledge.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524136EnglishN-0001November30General SciencesCLIMATE CHANGE AND DROUGHT HAZARD IN HILLS: A BURNING HYDRO-METEOROLOGICAL PROBLEM INVESTIGATED THROUGH REMOTE SENSING AND GIS MODELING
English116133Pradeep K. RawatEnglish P.C. TiwariEnglish Charu C. PantEnglishThe main objective of the study was to assess the impact of climate change on drought hazards through integrated Database Management System (DMS) on three GIS (Geographical Information System) modules i.e. climate informatics and land use-informatics and hydro-informatics. The Dabka watershed constitutes a part of the Kosi Basin in the Lesser Himalaya, India in district Nainital has been selected for the case illustration. Hydro-informatics module consists of daily, monthly and annual record of spring hydrology and stream hydrology of study area. Land use informatics consists of land use mapping and change diction i.e. decadal changes and annual changes whereas the Climate informatics module consists of spatial distribution of climate and its change detection through daily, monthly and annual weather data (temperature, rainfall, humidity and evaporation) of two study periods i.e. during 1985-1990 and 2005- 2010. The spatial distribution of climate throughout study area suggesting three types of climatic zones i.e. sub-tropical, temperate and moist temperate which are respectively favorable for mixed forest, pine forest and oak forest in the mountain eco-system. The results of climate-informatics advocating that all these climatic zones shifting towards higher altitudes due to global climate change and affecting the favorable conditions of the existing land use pattern. In order to that the oak and pine forests have decreased respectively by 25 % (4.48 km2) and 3% (.28 km2) during the period of 1990 to 2010. The
results also advocated that the overall accelerating factor of land use degradation in the study area broadly categorizes as dominant factor and supporting factor. Out of the total seven classes of the land use land cover, five classes (i.e. Oak, Pine, Mixed, Barren and Riverbed) are being degraded dominantly due to climate change factor and anthropogenic factors plays a supporting role whereas only two classes (Scrub land and agricultural land) are being change dominantly by anthropogenic factors and climate change factors plays a supporting role. Expansion of mixed forest land brought out due to upslope shifting of existing forest species due to climate change factor only because upslope areas getting warmer than past with the rate of 9?c-12?c/two decades. Consequently the results concluded that the high rate of land use degradation accelerating drought hazard during non-monsoon period as dry-up of natural springs and decreasing trends of stream discharge etc. This has serious implications on livelihood and food securities as natural springs and streams constitute the main source of drinking water and irrigation in the region.
EnglishClimate Change, Land use Degradation, Decreasing Water Resources, DroughtINTRODUCTION
Drought occurs over most parts of the world, even in wet and humid regions. It is a recurring extreme climate event over land characterized by below-normal precipitation over a period of months to years. This is because drought is defined as a dry spell elative on its local normal condition. On the other hand, arid areas are prone to drought because their rainfall amount ritically depends on a few rainfall events. Through out the world mainly three types of drought have been identified i.e. meteorological drought, agricultural drought and hydrological drought (Dai, 2011). Meteorological drought is a period of months to years with below-normal precipitation. It is often accompanied with above-normal temperatures, and precedes and causes other types of droughts. Agricultural drought is a period with dry soils that results from below-average precipitation, intense but less frequent rain events, or above-normal evaporation, all of which lead to reduced crop production and plant growth. Hydrological drought occurs when river stream flow and water storages in spring aquifers, lakes, or reservoirs fall below long-term mean levels. But the proposed study investigated that the Himalaya hills experiencing all three types of drought in same time period in a particular area due to climate change and land use degradation. In order to that the present study suggesting integrated approach of drought i.e. hydrometeorological drought which include all aspect of drought either it is agricultural drought, hydrological or meteorological drought because agriculture system in mountainous terrain completely depends on hydrological and meteorological parameters. A lack of precipitation often triggers hydro-meteorological droughts, but other factors, including more intense but less frequent precipitation, poor water management, and erosion, can also cause or enhance these droughts. Dwarfing all other mountains of the world in sheer height, Himalaya is the youngest mountain system, which is still undergoing tectonic movement due to prevailing geological conditions. Though each and every part of the world is more or less susceptible to natural calamities, the Himalaya due to its complex geological structures, dynamic geomorphology, and seasonality in hydro-meteorological conditions experience natural disasters very frequently, especially water-induced hazards (Rawat et. al., 2011). Climate change and land use degradation through flash flood, river-line flood, erosion, non-seismic land slide in monsoon period accelerating hydro-meteorological drought hazard as drying up of natural water springs and decreasing streams discharge in non-monsoon period particularly during summer season (April-June). The geodynamically active Himalayan terrain is being deforested at the rate of 0.36km2 /year (Rawat and Pant, 2007 and Rawat et al 2010). There are several anthropogenic factors that may contribute to this acceleration, including poorly managed agriculture, forest fire, overgrazing, and substandard construction of roads and buildings. Increasing population and demand of land for agriculture has resulted in pressure in the watershed of the Lesser Himalayan region (Rawat et al. 2011). This is further evident by upslope shifting and destruction of forest land due to climate change. An attempt has been made in the present study to assess the trends of land use pattern in a fragile watershed located near a seismic and tectonically active region of Himalya. The watershed lies between the latitude 29°24'09"– 29°30'19"E and longitude 79°17'53"-79°25'38"N in the north west of Nainital Township (Fig.1). The region encompasses a geographical area of 69.06 km2 between 700 m and 2623 m altitude above mean sea level. The total population of the watershed is 9250 people, which includes 16 Villages. The population density is 76.02 person /km2. . About 95% population of the total population depends on agriculture and forest resources but the forest cover is decreasing 0.36 km2 per year and the agricultural production also decreasing due climate change and drought hazard. The Geographical Information system (GIS) and Remote Sensing (RS) techniques have recently been widely applied to study land use/land cover changes (Mohanty, 1994; Minakshi et al., 1999; Brahmabhatt et al., 2000; and Chauhan et al., 2003). In Himalaya, a variety of changes have emerged in the traditional resource utilization structure mainly in response to population growth and resultant increased demand of natural resources, ineffective technology transfer, market forces, inappropriate land tenure policies, faulty environmental conservation programs, irrational rural developmental schemes, and increasing economic and political marginalization, during the recent years (Palni et al., 1998; Tiwari and Joshi, 1997). These emerging negative trends in the socio-economic profile have resulted into rapid exploitation and transformation of land resources and large-scale land use changes in the region (Tiwari, 1995). Under the impact of various land use systems, the land and whole environment of a geographical region changes positively or negatively. The impact of some land use changes is limited to the area in which they are operated while that of others reaches far in the surrounding ecosystems (Kostrowicki, 1983). The extensive land use changes in Himalaya have not only disrupted the fragile ecological balance of the watersheds in the region through deforestation, erosion, landslides, hydrological disruptions, depletion of genetic resources, but have also threatened the livelihood security and community sustainability in mountains as well as in adjoining plains ecosystem (Tiwari, 2000; Tiwari and Joshi, 2002 and 2005). The high rate of land use change accelerating several environmental problems such as high monsoon runoff, flash flood, river-line flood, soil erosion and denudation during monsoon season and drought during non-monsoon period as dry-up of natural springs and decreasing trends of stream discharge etc. These environmental problems cause great loss to life and property and poses serious threat to the process of development with have far-reaching economic and social consequences. Land use degradation due to climate change effecting water resources as drying up of natural water springs and decreasing trends of streams discharge and as a whole triggering other hydrological hazards such as high runoff, flash floods, river-line floods and non-seismic landslide etc. which are mainly responsible for several socio-economic consequences in mountainous terrain (Ives, 1989; Valdiya and Bartarya, 1989; Cruz, 1992; Jain et al, 1994; Sing, 2006; Rawat et. al., 2010). Flash flood in the region cause great loss to life and property and poses serious threat to the process of development with have far-reaching economic and social consequences. About hundreds of peoples are losing their life each year due to flash flood hazard in Himalaya region. On the other hand the river-line flood triggering several environmental socio-economic problems in many ways. River-line flood is undercutting of valley sides which causes the mountainsides to become unstable and ultimately ends in landslides and slope instability which demolished the infrastructural development (road network, buildings, canals, communication connectivity etc) and natural resources specially forest, land and water. During floods, tremendous amounts of erosion occurs on the banks of rivers and streams and washed away the crops and productive land whereas some time moreover, unsorted sediments are deposited over agricultural fields and settlements especially during monsoons.
METHODOLOGY
The study comprises mainly two components, (a) lab/desk study and (b) field investigations. The procedure adopted for study has been outlined in Fig. 2 which depicting that study carried out through GIS database management system (DMS). GIS-DMS is a set of computer programs for managing an integrated spatial and attribute database for such a task as map and data input storage, search, retrieval, manipulation and output. Existing DMS is constituted of three different GIS modules consisting of spatial map layers with their attribute data. These three GIS modules are: climate-informatics, land use-informatics and hydro-informatics respectively for climate change, land use degradation and drought hazard and its socio-economic and environmental risks assessment as describing below lines:
Climate Change Assessment
To assess the climate change during last twenty five years, a comprehensive meteorological study carried out for period 2005-2010 and compared the results of this study with previous study carried out during 1985-1990 (Bight, 1990) from the same study area (i.e. Dabka watershed). Consequentially the spatial distribution of climate throughout study area has been carried out as subtropical climate, temperate climate and moist temperate climate in respect to meteorological data of both study periods. These meteorological data recorded at five meteorological observatories. Four observatories located in sample micro-watershed established at different elevation and running by geology and geography department of Kumaun University and one located at lower elevation of the study area established and running by Irrigation Department of Uttarakhand state government. The four meteorological station running by Kumaun University funded by Govt. of India under different agencies as per their requirement i.e. Department of Environment (1985-1990), Department of Science and Technology (2005-2010) government of India. The meteorological records for a period of twenty five years (1985-2010) depicting that the lowest annual average temperature (18? c) found in year 1990 whereas year 2010 experienced highest annual average temperature (27? c) that‘s why those years determined as a base years to analysis the climate change and its impact on land use pattern which accelerating several ecological risks through high rate of runoff, rive line flood, flash flood and erosion etc. In order to that the elevation levels of the each meteorological observatory have been determined to delineate isotherms for year 1990 and 2010 and carryout spatial distribution maps of climate for respective year and than superimposed theses spatial distribution maps over the land use map for year 1990 and 2010 to detach the specific impact of climate change on the particular categories of land use land cover.
Land Use Land Cover Mapping and Change Detection
Indian Remote Sensing Satellite (IRS-1C) LISS III and PAN merged data of 1990 and 2010 was used for the analysis and mapping of land cover/ land use for the respective years (Fig. 2). Supplementary data and information required for the study have been generated from various primary as well as secondary sources. The primary information was generated through field surveys, mapping, interviews etc., and the relevant secondary data was collected from various sources, such as, Census of India – 2001, Government, Land Records, forests maps etc. Radiometric corrections were done employing dark pixel subtraction technique. The satellite images of the study area were registered geometrically using SOI Topographical Sheets (56 O/7NE and 56 O/7NW) of the area at scale 1:25000. For carrying out this important exercise uniformly distributed common Ground Control Points (GCPs) were selected and marked with root mean square (rms) error of one pixel and the images used were resampled by cubic convolution method. Both the data sets were then co-registered for further analysis initially, the LISS and PAN data were co-registered with root mean square (rms) error of 0.3 pixel and the output FCC was transformed into Intensity, Hue and Saturation (IHS) colour space images. The reverse transformation from IHS to RBG was performed substituting the original highresolution image for the intensity component, along with the hue and saturation components from the original RBG images. This merge data product obtained through the fusion of IRS –1C LISS – III and PAN was used for the generation of land cover/land use map of the study area for the year 2001, and digital image processing techniques supported by intensive ground truth surveys were used for the interpretation of the remote sensing data (Fig. 2). In order to enhance the interpretability of the remote sensing data for digital analysis several image enhancement techniques, such as, PCA, NDVI etc. were employed (Fig. 2). In the Himalayan mountain terrain the interpretability of the remote sensing data to a large extent is affected by the complexity of the terrain as due to the effect of elevation and slope and its aspect, the spectral signature of same objects are often different or vice versa. In order to overcome these constraints and also to attain the best possible level of accuracy in the interpretation, intensive ground truth surveys were carried out in the study region and a visual interpretation key was evolved for primary land cover/land use classification (Table 1). This was followed by the digital classification of land cover/ land use through on screen visual recording and rectification. To monitor the dynamics of land utilization pattern in the study area the land use maps generated for the years 1990 and 2010 were overlaid using Geographic Information System and land use changes were detected and mapped (Fig. 2). The geo-hydrological impacts of land use degradation due to climate change carried out for drought hazard and its risks appraisal (Rawat et. al., 2011).
Drought Hazard and Risks Appraisal
A comprehensive geo-hydrological study on spring hydrology and stream hydrology carried out during 2005-2010 and compared the results with the previous hydrological study (Bisht, 1990) to assess the impacts of climate change and land use degradation. Spring hydrology comprises of spatial distribution of different types of springs in respect to their discharge pattern (i.e. perennial non perennial and dried springs) and spring density. Stream hydrology comprises of spatial distribution of different types of stream (i.e. perennial non perennial and dried streams) and their density and frequency. The measurement techniques of spring and stream hydrology followed suggested by Rawat et al. 2011. In order to that a draught hazard and risks map carried out through integration of climate change, land use and hydrological database following scalogram modeling approach (Cruz, 1992).
RESULTS AND DISCUSSIONS
Climate-informatics The climate-informatics module consists of comparative study of climatic parameters for a twenty years period during 1990 to 2010 to appraise the impacts of climate change on land use pattern because it‘s reversely influences the ecology of the watershed and accelerated several environmental and socio-economic risks through high runoff, erosion and sediment delivery during rainy season. Monthly and annual climate data records of two study periods is used to climate spatial distribution mapping for respective year (Table 2 and Fig. 3). A brief account is as follows:
Temperature: The average temperature of the Dabka Watershed is 27ºC in present (2010) and it was about 18ºC twenty years back in 1990 although it varies altitude wise throughout the study area (Table 2 and Fig. 3). June is the driest and hottest month and receives highest annual temperature on the south-facing barren land whereas January is coldest month and receives minimum annual temperature. The annual maximum average temperature found 37ºC in 2010 whereas it was just about 23ºC twenty years back in 1990 (Table 2). The annual minimum average temperature recorded 12ºC in 1990 whereas it is has been rise up to 17ºC in present time by year 2010 (Table 2).
Rainfall: Throughout the watershed the average annual rainfall is about 2000 mm whereas twenty years back during 1985-1990 it was 2120 mm (Table 2). The results suggested that the annul rainfall pattern has been change and also experiencing that the rainfall season shirking by one to two months (August to September) with the characteristics of extreme rainfall within very small time period which becomes a big threat for hydrological hazards such as erosion, sediment delivery flood landslide and denudation etc. whereas the rainfall records by twenty year back depicting that the rainy season covered completely four months (June to September) having moderate rainfall pattern.
Humidity: The twenty year data analysis suggested that the average annual humidity of the watershed stands at 83.4% which varies between 86% in the dense oak forest area and 64.5% in the south-facing barren land. On other ecological conditions it stands at 80.7% in fairly dens forest/shrub land, 78.5% on the agricultural land agricultural land area. Table 2 suggesting that the average humidity was high than present it has been decreased by 8% annual due to climate change.
Evapotranspiration Loss: The twenty year data analysis suggested that the present annual evapotranspiration loss of the watershed is about 602.43 mm whereas twenty year back it was quit low about 535.57mm. Presently its approaches maximum up to 1072.3 mm on the south-facing barren land and drops down to 559.4 mm on the north-facing oak forest areas of the watershed. On other environmental conditions it was found 786.7 mm, 811.4 mm on the fairly dens forest/shrub land, agricultural land area respectively.
Spatial Distribution of Climate Change: The meteorological records for twenty five years (1985-2010) depicting that the lowest annual average temperature (18? c) found in year 1990 whereas year 2010 experienced highest annual average temperature (27? c) that‘s why those years determined as a base years to analysis the climate change and its impact on land use pattern which accelerating ecological degradation through deforestation, land degradation, instant rainfall, high monsoon runoff, flash floods, erosion and sediment delivery etc. (Rawat et al. 2011). The climate spatial distribution maps of two study periods (i.e. 1985-1990 and 2005-2010) advocating that all the existing climatic zones (i.e. sub-tropical, temperate and moist temperate) spreading towards higher altitudes due to global climate change and degrading the natural favorable climatic condition of the mixed forest, pine forest and oak forest i.e. respectively subtropical, temperate and moist temperate climate (Table 3 and Fig. 3). The results also advocating that the rates of the climate change are high in mountainous areas in compare to plain areas (Table 3 and Fig. 3). It means the rates of climate change are increasing in respect to higher elevation which dominantly effecting the natural vegetation covers throughout the Himalaya up to snow line (6000m). Above the snow line it‘s also affecting the snow cover which could be the major reason for malting of glaciers in the Himalaya which is a great threat to costal ecosystem due to rising of sea level. In order to monitor the impacts of climate change on land use pattern, the land use interpretation was carried out under land use-informatics module for the years 1990 and 2010 using IRS LISS–III and PAN merged data of respective years.
Land Use-informatics As mentioned in methodological section that the land use-informatics module consists of comparative land use land cover mapping for year 1990 and 2010 to assess the changes and identified the accelerating factors for the this changes as discussed in below.
Land Use for year 1990: On the basis of the above exercise, 7 land cover/land use classes could be identified in the study area, for 1990 and 2010 (Fig. 3). Besides the interpretation of the principal land use categories, all the major forest types available in the region could also be identified and mapped during the digital interpretation process. The land use/land cover maps of the study region have been prepared on the scale 1:25000 and classified into 7 categories for both the years (1990 and 2010). The land use classes identified in the watershed include (i) Oak Forests, (ii) Pine Forests, (iii) Mixed Forests, (iv) Scrub Land, (v) Barren Land, (vi) River Beds and Water Bodies, and (vii) Cultivated Land which also includes settlements (Fig. 3). In 1996, out of the total geographical area (69.06 km2 ) of Dabka Watershed as much as 39.23 km2 or 56.81% was under forests. The forests mainly include Oak Forests, Pine Forests and Mixed Forests, which respectively account for 26.05 % (17.99 km2 ), 11.89 % (8.21 km2 ), and 18.87 % (13.03 km2 ), of total geographical area of the watershed. The scrub land and barren land were respectively extended over 6.38 km2 (9.24 %) and 2.18 km2 (3.16 %) of the watershed area. Riverbeds and water bodies occupied only 1.50 km2 or 2.17 % geographical area of the study region. Nearly 20 km2 land surface which amounts to nearly 28.63 % of the total area of Dabka Watershed was under cultivation in 1990 (Table 3 and Fig. 3).
Land Use for year 2010: The forest emerged as the major land use/land cover also in the year 2010. A geographical area of 36.77 km2 , which accounts for nearly 53 % of total area of the watershed, has been classified as forests. Due to complexities of terrain and other geomorphic features the forests of the watershed are diversified in nature. Out of the total forest 22.20 % (15.33 km2 ) is under mixed forest, 19.56 % (13.51 km2 ) is under Oak forest, and 11.48 % (7.93 km2 ) is under Pine forests (Table 3). The hilly and mountainous parts of the watershed are covered with Oak and Pine species, whereas, in the lower elevations in the south mixed type of vegetation is very common. Agriculture and settlement are now confined to 20.40 km2 or 29.54 % of the total area. Scrub land, barren land and Riverbeds and water bodies respectively extend over 6.22 km2 (9.01 %), 3.39 km2 (4.91 %), 2.28 km2 (3.30 %) of the total geographical land surface of the study area (Table 3 and Fig. 3).
Land Use Change and Degradation during 1990-2010: In order to monitor the dynamics of land transformation process land use interpretation was carried out for the years 1990 and 2010 using IRS LISS – III and PAN merged data for the respective years. The exercise revealed that oak and pine forests have decreased respectively by 25 % (4.48 km2 ) and 3% (.28 km2 ) thus bringing a decline of 4.76 km2 forest in the watershed during 1990 to 2010. But, due to climate change the mixed forest taking place of oak forest in certain pockets and consequently the mixed forest in the catchment increased by 18 % (2.3 km2 ) during the same period which reduced the overall loss of forests in the region but its not eco-friendly as the oak forest because the broad leaved and wide spread roots of oak trees helps in controlling the several hydrological hazards such as accelerated runoff, erosion, landslides, flash flood and river-line flood during monsoon period and drought during non-monsoon period. As a result, the watershed recorded a total decline of 2.46 km2 or 6% forest area during 1990 to 2010 (Table 3 and Fig. 3). The non-forest area has increased dramatically due to lopping and cutting of trees, accelerated runoff, soil erosion, and growing agricultural activities. The non-forest area has mainly been confined to barren land, riverbed and cultivated land. Barren land increased 1.21 km2 (56%), riverbed increased 0.78 km2 (52%) and cultivated land increased about 0.63 km2 (3%) during the period of 1990 to 2010 (Table 3 and Fig. 3). The results of land use dynamics presented on Table 3, advocated that the overall accelerating factor of land use dynamics in the study area broadly categoriesed as dominant factor and supporting factor. Out of the total seven classes of the land use land cover, five classes (i.e. Oak, Pine, Mixed, Barren and Riverbed) are being change dominantly due to climate change factor and anthropogenic factors plays a supporting role whereas only two classes (Scrub land and agricultural land) are being change dominantly by anthropogenic factors and climate change factors plays a supporting role. Expansion of mixed forest land brought out due to upslope shifting of existing forest species due to climate change factor only because upslope areas getting warmer than past with the rate of 9?c - 12? c/two decades (Table 3 and Fig. 3).
Hydro-informatics
A comprehensive hydrological study on spring hydrology and stream hydrology carried out during 2005-2010 and compared the results with the previous hydrological study (Bisht, 1990) to assess the impacts of climate change and land use degradation. The results suggesting that the climate change accelerating land use degradation and drough hazard as decreasing under groundwater water level, drying up water springs and decreasing trends of stream discharge as discussed below:
Decreasing Underground Water Level: Land use degradation and deforestation reduced the protective vegetal cover as a result the significant proportion of rainfall goes waste as flood water without replenishing the groundwater reserve. It is experiencing that the ground water level throughout the watershed gradually going down due to deforestation and high flood runoff. The results advocating that twenty year back during 1985-1990 the underground water was easily approachable up to 2000 m altitude whereas now it is quit difficult due to deficiency of under ground water because the water table has been gone down up to just 1200 m altitude (Fig. 4a). The decreasing trends of under ground water table affecting the spring hydrology and stream hydrology in the study area as depicting by a schematic diagram in Fig. 4a for year 1990 and 2010.
Drying up Natural Water Springs: It was observed that, ?the springs are drying-up or being seasonal due to lack of ground water recharge in the catchment and the existing springs depicting decreasing water discharge rates (Table 4). This has serious implications on water resources and on livelihood and food securities as natural springs constitute the main source of drinking water and irrigation in the region. The spatial distribution of perennial spring carried out in the region revealed that there were total 116 perennial springs in the watershed in 1990 (Bisht, 1991), out of which 52% (60) have gone dry till 2010 (Table 4 and Fig. 4b, Fig. 4c). In order to that spatial distribution of perennial spring density has been carried out which depicting four categories of density i.e. low, moderate, high and very high. All these categories suggested the decreasing trend of spring density during 1990 to 2010 (Fig. 4d).
Decreasing Trends of Stream Discharge:
In order to drying up of natural springs the streams depicting decreasing trends of annual stream discharge and becoming perennial to seasonal streams in the study area. The hydrological results suggesting that the existing average annual discharge of Dabka watershed is 13 l/s/km2 whereas during 1985-1990 it was quit high i.e. 17 l/s/km2 (Table 4). It‘s also found that due to decreasing trends of stream discharge a number of perennial streams have been dried and as a result decreasing the perennial stream frequency throughout the sturdy area (Fig. 4b and Fig. 4c). The stream frequency map depicting four categories of the spatial distribution of perennial streams frequency (low, moderate, high and very high) and advocating that perennial stream frequency has been decreased during 1990 to 2010 due to climate change through drying up of natural water springs and decreasing trends of stream discharge (Fig. 4e).
Spatial Distribution of Drought Hazard: The spatial distribution of drought hazard comprises the integrated effect of above all three types of drought (i.e. decreasing underground water, drying up springs and streams) in a unit area. In order to that throughout the study area four zones of drought hazard have been determined i.e. low, moderate, high and very high in respect to potential for drought hazard. Very high hazard zone have the maximum area i.e. about 44.20 km2 or 64% of the watershed whereas very low hazard zone have minimum area (04.83 km2 or 7% of the watershed). The low and moderate drought hazard zones accounts respectively about 08.98 km2 or 13% and 11.05 km2 or 16% areas of the watershed (Table 5 and Fig. 5a).
Socio-Economic and Environmental Risks
Appraisal of Drought Hazard
To appraise the environmental and socioeconomic consequences of drought hazards due to climate change and land use degradation in the study area, a risks zone map has been prepared. This map comprises three zones of risk potential i.e. low, moderate and high potential. Areas mainly fall under forest, wasteland, channel beds and scrubland of the watershed having low socio-economic and environmental risks and accounts maximum area about 51.37 km2 or 74% of the study area (Table 5 and Fig. 5b). Areas located along streams, on alluvial and fluvial fans having about 10 % (7.06 km2 ) area of the watershed is under moderate risk potential zone whereas 15% (10.63 km2 ) area of the watershed under human settlements, water-mills, roads, canals and cultivated land, river terraces etc. having high risks potential for socio-economy and environment.
CONCLUSION
The GIS modeling on climate-informatics and land use-informatics advocated that the overall accelerating factor of land use degradation in the study area broadly categories as dominant factor and supporting factor. Out of the total seven classes of the land use land cover, five classes (i.e. Oak, Pine, Mixed, Barren and Riverbed) are being change dominantly due to climate change factor and anthropogenic factors plays a supporting role whereas only two classes (Scrub land and agricultural land) are being change dominantly by anthropogenic factors and climate change factors plays a supporting role. Expansion of mixed forest land brought out due to upslope shifting of existing forest species due to climate change factor only because upslope areas getting warmer than past with the rate of 9 ? c-12? c/two decades. Consequently the results concluded that the high rate of land use degradation accelerating drought hazard during non-monsoon period as dry-up of natural springs and decreasing trends of stream discharge etc.
ACKNOWLEDGEMENTS
This study constitutes part of multidisciplinary Collaborated project, on “Geo-environmental Appriasal of the Dabka Watershed, Kumaun Lesser Himalaya, District Nainital: A Model Study for Sustainable Development” funded to Authors by the NRDMS Division, DST, New Delhi. Dr. Pradeep Goswami, Senior Scientist, Center for climate change, Kumaun University Nainital helped in GIS analysis for which authors indebted to him. Thanks to Shri M.S. Bargali, project assistant helped during the intensive field work.
Fig. 3: Climate Zones for Year 1990 (a) and its Impact on Existing Land Use Pattern (a1), Climate Zones for Year 2010 and its Impact on Existing Land Use Pattern (b1)
Fig. 4: Geo-hydrological Impacts of Climate Change and Land Use Degradation During 1990-2010 in Himalaya i.e. Decreasing Under Ground Water Level and Dry up of Perennial Springs and Streams depicting by Schematic Diagram (a), Decreasing Perennial Springs and Streams during 1990 and 2010 (respectively b and c), Decreasing Perennial Springs Density (d) and Decreasing Perennial Streams Frequency (e),
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