Mirdul Yadav, Joyti Sethi, Kiran Dahyia, Sushma Sood, Veena Gupta, Veena Singh, Anjana Talwar

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1 experimental medicine EFFECT OF TRITICUM AESTIVIUM ON PHYSIOLOGICAL AND BIOCHEMICAL PARAMETERS IN HIGH FAT DIET FED RABBITS. Mirdul Yadav, Joyti Sethi, Kiran Dahyia, Sushma Sood, Veena Gupta, Veena Singh, Anjana Talwar Mridul Yadav, Assistant Professor, Jyoti Sethi Associate Professor,Sushma Sood, Prof.& Head Department of Physiology: Kiran Dahiya Department of Biochemistry, Veena Gupta Department of Pathology, Pt.B.D.Sharma University of Health Sciences, Rohtak, Haryana, India. Anjana Talwar, Associate Professor, Department of Physiology, AIIMS, Delhi, India. Received May 2011 Accepted May 2013 Correspondence: Dr. Mridul Yadav, Assistant Professor, Department of Physiology, Pt.B.D.Sharma University of Health Sciences, Rohtak, Haryana, India. Telephone No mridul There is always the need for the health adjuvants which, along with the conventional mode of treatment, can help to expediate the recovery from disease and to maintain the healthy state of body. Nowadays wheat grass and barley grass are gaining much popularity and claims about their health benefits range from providing supplemental nutrition to detoxification of liver and blood. The present studywas planned to assess the claims about health benefits of wheatgrass. Thirty rabbits were included in this study in group of ten each. Group i received control diet, group ii high fat diet and group iii high fat diet along with wheatgrass powder. The fasting blood sample analysis was done for various physiological and biochemical parameters. High fat diet induced derangement of liver functions (nsgot, nsgpt, ntotal protein and serum albumin) but the kidney functions remained unaffected. Wheat grass supplementation improved the hematological picture as well as liver function tests. Wheatgrass can be used as health improving adjuvant in liver disorders. Keywords: wheat grass, triticum aestivium, hyperlipidemia, oxidative stress, liver functiontests JK-Practitioner 2013;18(3-4):39-42 INTRODUCTION: The idea of grasses as foods with specific health benefits is nothing new. The cereal grasses (wheat grass, barley grass, rye grass, oat grass) have been used as human food supplements since 1930s. Laboratory research on the health benefits of cereal grasses has increased over the past two decades in the United States and Japan. 1 At the same time, the use of wheat grass as an alternative therapy for chronic diseases became popular. Wheat grass juice has been proven over many years to benefit people in numerous ways: cleansing the lymph system, building the blood, restoring balance in the body, removing toxic materials from the cells nourishing the liver and kidneys and restoring vitality. 2 Wheat grassjuice has been shown to reduce blood transfusion requirements in patients with beta thalassemia. 3 Wheatgrass 39 JK- Practitioner Vo 1.18, No (3-4) July-Dec

2 experimental medicine juice has been documented to be effective for an unrelated condition ulcerative colitis, in a randomi ed double-blind placebo-controlled trial from Israel. 4 Except for few current studies published on the role of wheat grass juice, most of the claims made in the literature are based on unpublished observations. Therefore, the present study was planned to evaluate the effect of wheatgrass on different physiological and biochemical parameters. MATERIAL AND METHODS Animals Albino rabbits of either sex weighing kg obtained from departmental animal house were used for this study. The rabbits were housed under controlled conditions of light (12 h light-dark cycle) and temperature (23 2) C with free access to respective diets and water ad libitum for a period of 10 weeks. Commercially available dried wheat grass powder (URJA-777) was used in the study. Institutional Animal Ethical Committee (I.A.E.C.) approval (IAEC/PATHO/08/ dated ) was obtained before the experiment and care was taken to handle the rabbits inhumane manner. Experimental design:- The animals were divided into the following three groups, with 10 rabbits in each group: Group I: (Control): Standard chow diet Group II: High fat diet (HFD) Group III: High fat diet plus wheat grass powder (2gm/day) The compositions of two diets were as follows 5 : Control Diet:- Wheat flour 22.5%, roasted Bengal gram powder 60%, skimmed milk powder 5%, casein 4%, refined oil 4%, salt mixture with starch 4% and vitamin and choline mixture 0.5%. High Fat diet:- Wheat flour 20.5%, roasted Bengal gram powder 52.6%, skimmed milk powder 5%, casein 4%, refined oil 4%, coconut oil 9%, salt mixture with starch 4%, vitamin and choline mixture 0.5% and cholesterol 0.4%. Body weight of animals of each group was recorded both before and after 10 week period. After 10 weeks, animals were kept fasting overnight and blood samples were collected from marginal vein of pinna. The serum was separated immediately and subjected to following biochemical estimations:- Hemoglobin (Hb) estimation was done by Sahli s acid hematinmethod. Red blood cell (RBC) count was done using haemocytometer. Packed cell volume (PCV) was done using wintrobe s tube. SGOT, SGPT, S. Protein, S. albumin, B. urea, S. creatinine estimation was done by autoanalyser Konelab 30 i using kits by Siemens and Randox. Statistical analysis: - Results were expressed as mean SD oflo rabbits in each group. The data was analysed statistically using SPSS software version 14.0 RESULTS There was no mortality during the ten JK- Practitioner Vo 1.18, No (3-4) July-Dec

3 experimental medicine weeks of experimental period and food intake was similar in all the three groups. The average body weight was found to have increased in high fat diet fed rabbits compared with those in control group. The physiological and biochemical parameters in three groups are shown in table 1. DISCUSSION Wheatgrass juice has been promoted as a supplementary health food/tonic for many years. Marwah et al have reported non-specific well being, improved appetite and reduced musculoskeletal aches and pains in thalassemics consuming wheat grassjuice. 3 Few recent studies have shown positive and promising results with wheat grass in the treatment of supprative wounds 6, hepatocellular carcinoma 7 and ulcerative colitis 4 but no definite mechanism has been able to explain its use in a wide variety of conditions. In group II rabbits, additional supplementation of high fat led to deranged liver functions as evident by increase in SGOT/PT and decrease in total proteins and serum albumin. However, kidney functions (blood urea and serum creatinine) were not affected by high fat diet. Rabbits receiving wheat grass supplementation showed normali ation of liver en ymes as well as that of total proteins and serum albumin. Mukhopadhyay et al have reported significant improvement (p.005) in mean value of Hb, total protein and serum albumin in terminally ill cancer patients receiving wheat grassjuice for 6 months. 8 Wheat grass juice contains 70% chlorophyll, chlorophyll being the basis of all plant life. 9 Wheat grass juice is a complete protein and contains amino acids arginine, serine, lysine, aspartic acid, glycine, alanine, methionine, tryptophan, phenyl alanine and valine. Among other en ymes present in wheatgrass vi - dnase, catalase, peroxidase, cytochrome oxidase, lipase; bioflavinoids and P4D1 require special mention. Bioflavinoids together with antioxidants offer a system of protection to the body. P4D1 is the first known substance to actually stimulate the DNA molecule and can regenerate the diseased cells. 10 Another component which can contribute to the above effects is water soluble Grass Juice Factor (GJF). 11 It is a powerful healing agent which functions as a systemic immunomodulator. Our study also suggests that the beneficial effect of wheatgrassjuice could be due to its rich nutritional content that includes Vitamins C and E and bioflavinoids. However gp III rabbits receiving wheatgrass supplementation showed a statistically significant increase in RBC count, Hb level and PCV. Wheatgrass juice has been reported to reduce transfusion requirements in patients with thalassemia major. Various mechanisms have been postulated to explain this. Some authors have attributed it to the very fact that chlorophyll (present in wheatgrass) and hemoglobin share a similar atom structure. The only difference in the two is that of metallic atom element. Hemoglobin consists of iron, while in chlorophyll the metallic atom is magnesium. 12 Fernandes and Donovan have speculated that the effects of wheat grass juice therapy may be due to the action of natural antioxidants on red blood cell 41 JK- Practitioner Vo 1.18, No (3-4) July-Dec

4 experimental medicine antioxidant function, and corresponding effects on cellular en yme function and membrane integrity. 13 Sethi et al have shown increase in level of antioxidants and decrease I in free radicals in serum of rabbits suggesting antioxidant potential of wheat grass. 14 Such an antioxidant mechanism of action of wheatgrass juice was also cited by Ben-Arye et al to explain the beneficial effects of wheatgrass juice therapy in patients with ulcerative colitis. 4 attributed the beneficial effect of wheatgrass juice to its rich nutritional content that includes Vitamins C and E and bioflavinoids. Our study also suggests that the improved physiological and biochemical picture could probably be due to inherent antioxidant potential of wheatgrass. However to substantiate our results, detailed phytochemical screening of Triticum aestivum grass as well as elucidation of bioactive compounds is needed. Conclusion Even though some encouraging results are shown by our study, the clinical effectiveness of wheat grass is not sufficiently defined at present. REFERENCES 1. Bing F., Secretary, AMA Council on Foods Accepted Foods Cerophyll. The Journal of the American Medical Association. 112: Wigmore, A The Wheatgrass Book. Avery Publishing Group.Wayne,NJ. 3. Marwah RK, Bansal D, Kaur S, Trehan A. Wheat grass juice reduces transfusion requirement in patients with Thalassemia major: a pilot study. Indian pediatr 2004; 41: Ben- Arye E, Goldin E, Wengrower D, Stamper A, Kohn R, Berry E. Wheatgrass juice in the treatment of active distal ulcerative colitis: Arandomi ed double blind placebo controlled trial. Scand J. Gastroenterol 2002; 37: Kottai MuthuA, Sethupathy S, Manavalan R, Karar PK. Antioxidant potential of methanolic extract of Dolichos biflorus Linn in high fat diet fed rabbits. Indian J Pharmacol 2006;38: Gruskin B. Chlorophyll- its therapeutic place in acute and suppurative disease. Am J Surg 1940;49: Egner PA, Muno A, Kensler TW. Chemoprevention with chlorophyllin in individuals exposed to dietary aflatoxin. Mutat Res 2003; Feb-March : Mukhopadhyay S, Sonali D, Gupta PR, Mukhopadhyay A. Effect of wheatgrassjuice in supportive care of terminally ill solid organ cancer patients: experience from eastern India. Netaji Subhash Chandra Bose cancer research institute Kolkata India. Study from Jan 2003 to Dec Patek, A.. Chlorophyll and regeneration of blood. Archives of Internal Medicine.1936; 57: Kubota, K. and Matsuoka, Y. Isolation of potent anti-inflammatory protein from Barley leaves. Japanese Journal of Inflammation.1983; 4: Kohler G. Elvehjem, C. Hart F. Growth stimulating properties of grassjuice. Science. M1936; Gala D, Gala DR, Gala S, Chlorophyll; A potent prophylactic and Curative substance. In: Panacea on the Earth-Wheat grass juice. Navneet Publications ( India) Limited, Dantali, Gujarat. 2000; pp Fernandes CJ, O Donovan DJ. Natural antioxidant therapy for patients with hemolytic anaemia. Indian pediatr 2005; 42: Sethi J, Yadav M, Dahiya K, Sood S, SinghV, Bhattacharya S. Antioxidant effect of Triticum 15. Aestium (wheat grass) in high fat diet induced oxidative stress in rabbits.methods and Findings in Experimental and Clinical Pharmacology. 2010;32: JK- Practitioner Vo 1.18, No (3-4) July-Dec

5 Journal Scan Journal Scan 1 Ann Intern Med Dec 18;157(12): Probiotics for the prevention of Clostridium difficile-associated diarrhea: a systematic review and meta-analysis. Johnston BC, Ma SS, Goldenberg JZ, Thorlund K, Vandvik PO, Loeb M, Guyatt GH. Author information The Hospital for Sick Children Research Institute, Room 2420, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada. bradley.johnston@sickkids.ca BACKGROUND: Antibiotic treatment may disturb the resistance of gastrointestinal flora to colonization. This may result in complications, the most serious of which is Clostridium difficileassociated diarrhea (CDAD). PURPOSE: To assess the efficacy and safety of probiotics for the prevention of CDAD in adults and children receiving antibiotics. DATA SOURCES: Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, CINAHL, Allied and Complementary Medicine Database, Web of Science, and 12 grayliterature sources. STUDYSELECTION: Randomized, controlled trials including adult or pediatric patients receiving antibiotics that compared any strain or dose of a specified probioticwith placebo or with no treatment control and reported the incidence of CDAD. DATA EXTRACTION: Two reviewers independently screened potentially eligible articles; extracted data on populations, interventions, and outcomes; and assessed risk of bias. The Grading of Recommendations Assessment, Development and Evaluation guidelines were used to independently rate overall confidence in effect estimates for each outcome. DATA SYNTHESIS: Twenty trials including 3818 participants met the eligibility criteria. Probiotics reduced the incidence of CDAD by 66% (pooled relative risk, 0.34 [95% CI, 0.24 to 0.49]; l(2) = 0%). In a population with a 5% incidence of antibiotic-associated CDAD (median control group risk), probiotic prophylaxis would prevent 33 episodes (CI, 25 to 38 episodes) per 1000 persons. Of probiotic-treated patients, 9.3% experienced adverse events, compared with 12.6% of control patients (relative risk, 0.82 [CI, 0.65 to 1.05]; l(2) = 17%). LIMITATIONS: In 13 trials, data on CDAD were missing for 5% to 45% of patients. The results were robust to worst-plausible assumptions regarding event rates in studies with missing outcome data. CONCLUSION: Moderate-quality evidence suggests that probiotic prophylaxis results in a large reduction in CDAD without an increase in clinically important adverse events. PRIMARY FUNDING SOURCE: None. JK- Practitioner Vo 1.18, No (3-4) July-Dec

6 Journal Scan Journal Scan 2 BMJ Jan 28:346:f360. Efficacy and safety of dual blockade of the renin-angiotensin system: meta-analysis of randomised trials. Makani H. Bangalore S. Desouza KA. ShahA, Messerli FH. Author information Division of Cardiology, St Luke's Roosevelt Hospital, Columbia University College of Physicians and Surgeons, th Avenue, New York, NY 10019, USA. OBJECTIVE: To compare the long term efficacy and adverse events of dual blockade of the renin-angiotensin system with monotherapy. DESIGN: Systematic review and meta-analysis. DATA SOURCES: PubMed, Embase, and the Cochrane central register of controlled trials, January 1990 to August2012. STUDYSELECTION: Randomised controlled trials comparing dual blockers of the renin-angiotensin system with monotherapy, reporting data on either long term efficacy ( 1 year) or safety events ( 4 weeks), and with a sample size of at least 50. Analysis was stratified by trials with patients with heart failure versus patients without heart failure. RESULTS: 33 randomised controlled trials with 68,405 patients (mean age 61 years, 71% men) and mean duration of 52 weeks were included. Dual blockade of the reninangiotensin system was not associated with any significant benefit for all cause mortality (relative risk 0.97, 95% confidence interval 0.89 to 1.06) and cardiovascular mortality (0.96, 0.88 to 1.05) compared with monotherapy. Compared with monotherapy, dual therapy was associated with an 18% reduction in admissions to hospital for heart failure (0.82, 0.74 to 0.92). However, compared with monotherapy, dual therapy was associated with a 55% increase in the risk of hyperkalaemia (P 0.001), a 66% increase in the risk of hypotension (P 0.001), a 41% increase in the risk of renal failure (P=0.01), and a 27% increase in the risk of withdrawal owing to adverse events (P 0.001). Efficacy and safety results were consistent in cohorts with and without heart failure when dual therapy was compared with monotherapy except for all cause mortality, which was higher in the cohort without heart failure (P=0.04 v P=0.15), and renal failure was significantly higher in the cohort with heart failure (P v P=0.79). CONCLUSION: Although dual blockade ofthe renin-angiotensin system may have seemingly beneficial effects on certain surrogate endpoints, itfailed to reduce mortality and was associated with an excessive risk of adverse events such as hyperkalaemia, hypotension, and renal failure compared with monotherapy. The risk to benefit ratio argues against the use of dual therapy. JK- Practitioner Vo 1.18, No (3-4) July-Dec

7 Journal Scan Journal Scan 3 Chest Oct:144(4): Risk of serious atrial fibrillation and stroke with use of bisphosphonates: evidence from a meta-analysis. SharmaA, Chatterjee S. Arbab-ZadehA, GovalS, Lichstein E. Ghosh J. AikatS. BACKGROUND: Clinical studies have suggested an association between bisphosphonate use and the onset of atrial fibrillation (AF). However, data on the risk of developing AF, stroke, and cardiovascular mortality with the use of bisphosphonate are conflicting. The objective of this study was to evaluate the risk of serious AF (events that required hospital admission), stroke, and cardiovascular mortality with the use of bisphosphonates through a systematic review of the literature. METHODS: We searched the PubMed, CENTRAL, and EMBASE databases for observational studies and randomized controlled trials (RCTs) on the use of bisphosphonates from 1966 to April 2012 that reported the number of patients who developed serious AF, stroke, and cardiovascular mortality at follow-up. The random-effects Mantel-Haenszel test was used to evaluate relative riskadverse cardiovascular outcomes with the use of bisphosphonates. RESULTS: Six observational studies (n = 149,856) and six RCTs (n =41,375) were included for analysis. On pooling observational studies, there was an increased risk of AF (OR, 1.27; 95% CI, ) among bisphosphonate users. Further, analysis of RCTs revealed a statistically significant increase in the risk of serious AF (OR, 1.40; 95% CI, ) and no increase in the risk of stroke (OR, 1.07; 95% CI, ) or cardiovascular mortality (OR, 0.92; 95% CI, ) with the use of bisphosphonates. CONCLUSIONS: Evidence from RCTs and observational studies suggests a significantly increased risk of AF requiring hospitalization, but no increase in risk of stroke or cardiovascular mortality, with the use of bisphosphonate. PMID: [PubMed - indexed for MEDLINE] JK- Practitioner Vo 1.18, No (3-4) July-Dec

8 Journal Scan Journal Scan 4 Cochrane Database Syst Rev Sep 13:9:CD Holding chambers (spacers) versus nebulisers for beta-agonist treatment of acute asthma. CatesCJ. Welsh EJ. RoweBH. Author information Population Health Sciences and Education, St George's University of London, CranmerTerrace, London, UK, SW17 ORE. BACKGROUND: In acute asthma inhaled beta(2)-agonists are often administered by nebuliser to relieve bronchospasm, but some have argued that metered-dose inhalers with a holding chamber (spacer) can be equally effective. Nebulisers require a powersource and need regular maintenance, and are more expensive in the community setting. OBJECTIVES: To assess the effects of holding chambers (spacers) compared to nebulisers for the delivery of beta(2)-agonists for acute asthma. SEARCH METHODS: We searched the Cochrane Airways Group Trial Register and reference lists of articles. We contacted the authors of studies to identify additional trials. Date of last search: February SELECTION CRITERIA: Randomised trials in adults and children (from two years of age) with asthma, where spacer beta(2)-agonist delivery was compared with wetnebulisation. DATA COLLECTION AND ANALYSIS: Two review authors independently applied study inclusion criteria (one review author for the first version of the review), extracted the data and assessed risks of bias. Missing data were obtained from the authors or estimated. Results are reported with 95% confidence intervals (CIs). MAIN RESULTS: This review includes a total of 1897 children and 729 adults in 39 trials. Thirty-three trials were conducted in the emergency room and equivalent community settings, and six trials were on inpatients with acute asthma (207 children and 28 adults). The method of delivery of beta(2)-agonist did not show a significant difference in hospital admission rates. In adults, the risk ratio (RR) of admission for spacer versus nebuliser was 0.94 (95% CI 0.61 to 1.43). The risk ratiofor children was 0.71 (95% CI 0.47 to 1.08, moderate quality evidence). In children, length of stay in the emergency department was significantly shorter when the spacer was used. The mean duration in the emergency department for children given nebulised treatment was 103 minutes, and for children given treatment via spacers 33 minutes less (95% CI -43 to -24 minutes, moderate quality evidence). Length of stay in the emergency department for adults was similar for the two delivery methods. Peak flow and forced expiratory volume were also similar for the two delivery methods. Pulse rate was lower for spacer in children, mean difference- 5% baseline (95% CI -8% to -2%, moderate quality evidence), as was the risk of developing tremor (RR 0.64; 95% CI 0.44 to 0.95, moderate quality evidence). AUTHORS' CONCLUSIONS: Nebuliser delivery produced outcomes that were not significantly better than metered-dose inhalers delivered by spacer in adults or children, in trials where treatments were repeated and titrated to the response ofthe participant. Spacers may have some advantages compared to nebulisers for children with acute asthma. 43 JK- Practitioner Vo 1.18, No (3-4) July-Dec

Wheat Grass Juice Reduces Transfusion Requirement in Patients with Thalassemia Major: A Pilot Study

Wheat Grass Juice Reduces Transfusion Requirement in Patients with Thalassemia Major: A Pilot Study R.K. Marwaha, Deepak Bansal, Siftinder Kaur and Amita Trehan From the Division of Pediatric Hematology-Oncology,