A randomized, controlled, comparative, proof-of-concept study to evaluate the efficacy and safety of Nisha-Amalaki capsules in prediabetic patients for preventing progression to diabetes.

BACKGROUND: Prediabetes is an intermediate state of hyperglycemia, which acts as a precursor to Diabetes mellitus if left untreated. Nisha (Curcuma longa) and Amalaki (Emblica officinalis) combination has been advocated as drugs of choice to treat the early manifestations of Diabetes mellitus. OBJECTIVE: This prospective, randomized, single-blind, placebo-controlled, comparative study was planned to assess the efficacy and safety of Nisha-Amalaki capsules in preventing progression to Diabetes mellitus in prediabetic patients when administered for 6 months. METHODS: The study was conducted on prediabetic participants randomized to receive either Nisha-Amalaki (500 mg) or placebo one capsule twice a day for six months. The effect of study medications on IDRS (Indian Diabetes Risk Score), BMI (Body Mass Index), blood sugar, serum insulin, HOMA-IR (Homeostasis Model Assessment-Estimated Insulin Resistance), HbA1c (glycated hemoglobin), oxidative markers, Ayurvedic symptoms and Quality of Life (QoL) scores was assessed at regular intervals. RESULTS: 58 of the 62 participants enrolled completed the study. Significant fall in IDRS score [p < 0.001], BMI [p < 0.001], fasting, and 2 h post-OGTT sugar, insulin, HbA1c, HOMA-IR, and oxidative stress markers [p < 0.001] was observed in patients receiving Nisha-Amalaki at 6 months. Ayurvedic symptoms and QoL scores also improved at 6 months in the treatment group. CONCLUSION: Treatment with Nisha-Amalaki capsules improved all study parameters including insulin sensitivity at 6 months as compared to placebo in prediabetic patients. Thus Nisha-Amalaki should be considered as prophylactic therapy in prediabetics to delay progression to diabetes.

pH-responsive microparticles of rifampicin for augmented intramacrophage uptake and enhanced antitubercular efficacy.

In this study we present pH-responsive rifampicin (RIF) microparticles comprising lecithin and a biodegradable hydrophobic polymer, polyethylene sebacate (PES), to achieve high intramacrophage delivery and enhanced antitubercular efficacy. PES and PES-lecithin combination microparticles (PL MPs) prepared by single step precipitation revealed average size of 1.5 to 2.7 µm, entrapment efficiency âˆ¼ 60 %, drug loading 12-15 % and negative zeta potential. Increase in lecithin concentration enhanced hydrophilicity. PES MPs demonstrated faster release in simulated lung fluid pH 7.4, while lecithin MPs facilitated faster and concentration dependent release in acidic artificial lysosomal fluid (ALF) pH 4.5 due to swelling and destabilization confirmed by TEM. PES and PL (1:2) MPs exhibited comparable macrophage uptake which was âˆ¼ 5-fold superior than free RIF, in the RAW 264.7 macrophage cells. Confocal microscopy depicted intensified accumulation of the MPs in the lysosomal compartment, with augmented release of coumarin dye from the PL MPs, confirming pH-triggered increased intracellular release. Although, PES MPs and PL (1:2) MPs displayed comparable and high macrophage uptake, antitubercular efficacy against macrophage internalised M. tuberculosis was significantly higher with PL (1:2) MPs. This suggested great promise of the pH-sensitive PL (1:2) MPs for enhanced antitubercular efficacy.

An open labeled, randomized, controlled clinical study to evaluate the efficacy of Torchnil capsules and Febcin tablet as add-on therapy for Covid-19 patients.

Objectives: This study was conducted to evaluate the efficacy and safety of Cap. Torchnil & Tab. Febcin when given as add-on therapy to Covid19 positive patients with moderate disease. Material and methods: Following written informed consent, patients were randomized to receive Cap. Torchnil & Tab. Febcin in addition to standard of care (SOC) [Add-on Group] or only SOC [SOC Group] for 14 days. Effect on clinical symptoms, WHO Clinical Assessment scale, hospital stay duration, time to Covid negative report, Sp02 levels and biomarkers was assessed during admission and relapse rate, if any, post discharge for 3 months. Results: 193 patients were screened and 150 completed the study, 77 in Add-on Group and 73 in SOC Group. Improvement in Covid related symptoms, WHO Assessment scale, time to covid negative report and duration of hospital stay was observed earlier in Add-on Group. Statistically significant fall in biomarker levels viz. CPK, D-dimer and IL-6 values at Day 14 and LDH levels at Days 7 & 14 was observed in Add-on Group. Improvement in Sp02 levels was also seen earlier in Add-on Group. Only 2 patients complained of acidity. Post discharge, 91 patients (49 from Add-on group and 42 from SOC group) came for physical visits. All these patients were clinically stable with no evidence of relapse. Conclusion: The study results thus showed that Cap. Torchnil and Tab. Febcin were effective and safe when given as add-on therapy to SOC in the clinical management of patients with moderate Covid-19 disease.

In-Vitro Evaluation of Antimicrobial Activities of Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Neisseria gonorrhoeae, and Candida albicans Nosodes.

BACKGROUND: This study presents the results of the minimum inhibitory concentration (MIC) assay of a series of nosodes: namely Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Neisseria gonorrhoeae, and Candida albicans. Each was tested against its corresponding infection as well as cross infections. METHODS: In-vitro efficacy of polyvalent nosodes was tested using the MIC assay technique. The nosodes, namely C. albicans polyvalent nosode (35c, 100c), N. gonorrhoeae (35c), K. pneumoniae (35c, 100c), E. coli polyvalent nosode (35c, 100c) and Salmonella typhi polyvalent nosode (30c, 100c), were tested along with positive and negative controls. Nosodes were studied in different potencies and at 1:1 dilution. RESULTS: C. albicans polyvalent nosode 35c, 100c, N. gonorrhoeae 35c, and positive control amphotericin B showed inhibition of the growth of C. albicans species. K. pneumoniae 35c, E. coli polyvalent nosode 100c, and meropenem (positive control) showed inhibition of the growth of K. pneumoniae; this effect was not seen with ceftriaxone, ofloxacin and amoxicillin antibiotics. E. coli polyvalent nosode 30c in 10% alcohol (direct and dilution 1:1) and the positive controls ciprofloxacin, ofloxacin, and amoxicillin showed inhibition of the growth of E. coli. The S. typhi polyvalent nosode 30c in 10% alcohol showed inhibition of growth of S. typhi. CONCLUSION: This study reveals that the tested nosodes exhibited antibacterial potential against the corresponding micro-organisms and against other selected organisms studied using this assay.

Preparation and Standardization of Nosodes Sourced from Klebsiella Pneumoniae, Salmonella Typhi, Neisseria Gonorrhoeae and Candida Albicans Strains.

BACKGROUND: Homeopathic nosodes prepared from organisms and pathological tissues have shown biological effects, encouraging more research. There is a need to develop some new nosodes systematically and to re-make others that were developed over a century ago. In our program of work on nosodes, the bacterial strains Klebsiella pneumoniae (BAA 2146), Salmonella typhi and Neisseria gonorrhoeae (ATCC 43069), and the single-celled fungus Candida albicans (24433, 26790, and 60193) have been identified for preparation. MATERIALS AND METHODS: The systematic and scientific method of preparation of nosodes includes identification, culture, quantification, characterization, preparation, and standardization. Under laminar flow, a suspension of respective bacterial and fungal cells (20 billion cells/mL) was processed as per the Homoeopathic Pharmacopoeia of India (HPI). Culture tests, sterility tests and molecular testing (polymerase chain reaction) were performed to establish the absence of contamination, live organisms and DNA material. RESULTS: K. pneumoniae, S. typhi (single, bivalent, or polyvalent), N. gonorrhoeae, and C. albicans nosodes (single and polyvalent) were sourced and prepared from different strains of respective cultures. The nosode preparations were processed by serial dilution and potentization, normally following the HPI guidelines. Molecular test results showed the absence of live organisms or DNA material; culture and sterility test results demonstrated the safety profile of the potentized nosodes. CONCLUSION: K. pneumoniae, S. typhi, N. gonorrhoeae and C. albicans nosodes were successfully prepared. Their therapeutic potential may now be evaluated.

A Phase 3, Randomized, Open-label, Noninferiority Trial Evaluating Anti-Rabies Monoclonal Antibody Cocktail (TwinrabTM) Against Human Rabies Immunoglobulin (HRIG).

BACKGROUND: Limited supply, cost and potential for severe adverse effects observed with the blood derived rabies immunoglobulin products has led to search for alternative therapies. This issue has been addressed by developing an anti-rabies monoclonal antibody cocktail. METHODS: This is a phase 3, randomized, open-label, noninferiority trial conducted in patients with World Health Organization (WHO) category III exposure with suspected rabid animal. Eligible patients were assigned to either the test arm, TwinrabTM (docaravimab and miromavimab) or the reference arm, human rabies immunoglobulin (HRIG; Imogam® Rabies-HT), in a ratio of 1:1. The primary endpoint was the comparison of responder rates between the 2 arms assessed as percentage of those with rabies virus neutralizing antibodies titers ≥0.5 IU/mL on day 14. RESULTS: A total of 308 patients were equally randomized into the 2 arms. In the per-protocol (PP) population, there were 90.21% responders in the TwinrabTM arm and 94.37% in the HRIG arm. The geometric mean of rapid fluorescent foci inhibition test titers in the PP on day 14 were 4.38 and 4.85 IU/mL, for the TwinrabTM and HRIG arms, respectively. There were no deaths or serious adverse events reported. CONCLUSIONS: This study confirmed that TwinrabTM is noninferior to HRIG in terms of providing an unbroken window of protection up to day 84. This trial in healthy adults with WHO category III exposure from suspected rabid animal also establishes the safety of TwinrabTM in patients with 1 WHO approved vaccine regimen (Essen). CLINICAL TRIALS REGISTRATION: CTRI/2017/07/009038.

Does Panchatikta ghrita have anti-osteoporotic effect? Assessment in an experimental model in ovariectomized rats.

BACKGROUND: Osteoporosis is a public health problem in the elderly wherein a decrease in bone mass and mineral density increases the at risk of fractures. Panchatikta Ghrita (PG) is a classical Ayurvedic formulation that may help slow bone degeneration. OBJECTIVE: This experimental study was conducted to assess the efficacy of Panchatikta ghrita (PG) in protecting against postmenopausal osteoporosis in ovariectomized rats. MATERIALS AND METHODS: The experiment was initiated after Institutional Animal Ethics Committee approval. 96 female Sprague Dawley rats were divided into 8 groups viz. sham control (NC), diseased control (DC), vehicle control (VC), 3 test drug (PG) groups (PG1, PG2 & PG3 - 0.9, 1.8 and 2.7gm/kg body weight respectively) and 2 standard control (SC) groups - SC1 received 17α-ethinylestradiol 1µg/kg/day while SC2 received alendronate (7mg/kg/week). Study medications were administereddaily for four months. Bone specific biomarkers viz. osteocalcin and TRAP-5b were estimated at baseline and end of study. Animals were sacrificed on day 121 and their femurs and tibiae were harvested for histomorphometric analysisand bone microarchitectural studies. RESULTS: Serum osteocalcin and TRAP-5b showed significant increase (p < 0.001) in levels in DC group as compared to sham controls. All 3 doses of PG decreased bone specific biomarker levels with maximal effect seen with highest dose of PG similar to that seen with standard drugs. PG also significantly improved bone micro architectural parameters like bone mineral density and mineral content at higher dose levels. Decrease in osteoclasts and significant dose dependent increase in bone hardness and elasticity was seen with PG which was comparable to standard drugs. CONCLUSION: PG increased bone mineral density and content, decreased turnover of bone specific biomarkers and osteoclast formation, indicating its protective effect against experimentally induced postmenopausal osteoporosis.

Chrysin mitigated obesity by regulating energy intake and expenditure in rats.

BACKGROUND AND AIM: Chrysin is a flavonoid found in plant extracts from Passiflora species, honey and propolis. It has demonstrated anti-adipogenic activity in vitro but there are no studies substantiating the anti-obesity activity of chrysin in vivo. EXPERIMENTAL PROCEDURE: The pancreatic lipase (PL) inhibitory potential of chrysin was determined by preliminary in silico screening and further confirmed by in vitro PL inhibitory assay and oral fat tolerance test (OFTT). The effect of chrysin on acute feed intake and sucrose preference test was determined in normal rats. Obesity was induced by feeding of high fructose diet (HFD) to the rats. The rats were divided into six groups: normal control, HFD control, orlistat and three doses of chrysin (25, 50 and 100 mg/kg body weight). Body weight, body mass index (BMI), abdominal circumference/thoracic circumference (AC/TC) ratio, calorie intake, adiposity index, fecal cholesterol, locomotor activity and histopathology of the adipose tissue of the rats were evaluated. RESULTS: Chrysin showed good affinity to PL with competitive type of inhibition. It significantly reduced serum triglycerides in OFTT. Chrysin also significantly reduced acute feed intake and sucrose preference in rats. Chrysin significantly decreased the body weight, BMI, AC/TC ratio, adiposity index, calorie intake while it significantly increased the fecal cholesterol and locomotor activity of the rats. Chrysin was found to reduce the size of the adipocytes when compared to the HFD control group. CONCLUSION: Thus, chrysin exerted anti-obesity effect by inhibiting PL, reducing sucrose preference, reducing calorie intake and increasing the locomotor activity of rats.

Preparation and Standardization of Escherichia coli Nosodes Sourced from Select E. coli Strains.

BACKGROUND: The nosodes are well-known preparations in homeopathy that are sourced from organisms and diseased materials. More than 40 known nosodes have been used in homeopathic practice for over a century. Having identified the need for scientifically developed new nosodes sourced from organisms that are currently prevalent, the preparation of Escherichia coli nosodes from different strains of the bacterium is presented in this article. MATERIALS AND METHODS: Escherichia coli strains (E. coli ATCC 11775E, ATCC 25922, and ATCC 8739) were identified, cultured, and tested for purity, and 20 billion cells were processed following the nosode preparation method given in the Homoeopathic Pharmacopoeia of India, group N1. Serial dilution and potentization for liquid potency were done up to 30c potency. Nosodes were prepared by two methods: from cell-free extract (endotoxin) and from entire-cell extract. RESULT: Six nosodes were developed in total. Three univalent nosodes were prepared using individual endotoxins, one from each of the three E. coli strains; those three univalent nosodes were also combined as "Trivalent nosode-I". "Trivalent nosode-II" was prepared by mixing entire cells of the three E. coli strains. A mix of both Trivalent nosode-I and Trivalent nosode-II was labeled "EC-Polynosode". The safety profile of the potentized nosodes was documented by the non-detectability of traces of source material (absence of contamination, live organisms, or DNA material) through a culture test, sterility test, and molecular testing (polymerase chain reaction). CONCLUSION: Different variants of E. coli nosodes were systematically and scientifically prepared and standardized using the cultures. Homeopathic pathogenetic trials, in-vitro efficacy studies, and clinical evaluation of E. coli nosodes (single, trivalent, or polyvalent nosodes) will be required in future.

Punica granatum improves renal function in gentamicin-induced nephropathy in rats via attenuation of oxidative stress.

BACKGROUND: Gentamicin is widely used as an antibiotic for the treatment of gram negative infections. Evidences indicates that oxidative stress is involved in gentamicin-induced nephrotoxicity. In Ayurvedic medicine, Punica granatum Linn. is considered as 'a pharmacy unto itself". It has been claimed in traditional literature, to treat various kidney ailments due to its antioxidant potential. OBJECTIVE: To explore the possible mechanism of action of methanolic extract of P.granatum leaves (MPGL) in exerting a protective effect on gentamicin-induced nephropathy. MATERIAL AND METHODS: Animals were administered with gentamicin (80 mg/kg/day i.m.) and simultaneously with MPGL (100, 200 and 400 mg/kg p.o.) or metformin (100 mg/kg p.o.) for 8 days. A satellite group was employed in order to check for reversibility of nephrotoxic effects post discontinuation of gentamicin administration. At the end of the study, all the rats were sacrificed and serum-urine parameters were investigated. Antioxidant enzymes and tumor necrosis factor alpha (TNF-α) levels were determined in the kidney tissues along with histopathological examination of kidneys. RESULTS: Increase in serum creatinine, urea, TNF-α, lipid peroxidation along with fall in the antioxidant enzymes activity and degeneration of tubules, arterioles as revealed by histopathological examination confirmed the manifestation of nephrotoxicity caused due to gentamicin. Simultaneous administration of MPGL and gentamicin protected kidneys against nephrotoxic effects of gentamicin as evidenced from normalization of renal function parameters and amelioration of histopathological changes. CONCLUSION: Data suggests that MPGL attenuated oxidative stress associated renal injury by preserving antioxidant enzymes, reducing lipid peroxidation and inhibiting inflammatory mediators such as TNF-α.

Assessing the Effectiveness of Panchatikta Ghrita, a Classical Ayurvedic Formulation as Add-on Therapy to Vitamin D3 and Calcium Supplements in Patients with Osteopenia: A Randomized, Open-Labeled, Comparative, Controlled Clinical Study.

Objectives: To assess the change in the bone mineral density (BMD) score, bone-specific biomarkers (serum vitamin D3, tartrate-resistant acid phosphatase 5b [TRAP-5b], and osteocalcin), quality of life, Ayurvedic symptoms (Asthikshaya Lakshanas), and fracture risk assessment tool (FRAX) scores following treatment with Panchatikta Ghrita (PG), a classical herbal formulation as add-on therapy to calcium and vitamin D3 supplements. Study design: Randomized, open-labeled, comparative, controlled clinical study. Location: TN Medical College and BYL Nair Hospital, Mumbai, India. Study participants: Eighty adult patients, aged between 40 and 75 years, diagnosed to have osteopenia (BMD T-score between -1 and -2.5 in at least two of the three joints tested-lumbar spine L1-L4, left femur-neck, left forearm-radius total). Study intervention: Treatment group received two tablespoons of PG (10 mL in lukewarm milk) along with calcium and vitamin D3 supplements twice a day, whereas control group received only calcium and vitamin D3 supplements twice a day for a period of 12 months. Outcome measures: BMD, bone-specific biomarkers (vitamin D3, TRAP-5b, and osteocalcin), quality of life, Ayurvedic symptoms, and FRAX scores were evaluated before and at 6 and 12 months. Results: Eighty patients were enrolled; of which, 65 patients completed the study while 15 patients dropped out. Improvement in the BMD scores was observed at 6 and 12 months with the maximum benefit in the lumbar spine region. Significant improvement in the bone-specific biomarkers, namely serum vitamin D3 (p < 0.001), osteocalcin (p < 0.001), and TRAP-5b (p < 0.05), was observed in the PG-treated group compared with the standard treatment group. Improvement in the quality of life, Ayurvedic symptoms scores, and risk reduction in FRAX scores of major osteoporotic fracture risk and hip fracture risk was greater with PG, although not statistically significant. Conclusions: The study findings demonstrate that PG slows down the bone degeneration processes by its stabilizing effect on the bone-specific biomarkers, indicating its potential usefulness as preventive therapy in osteopenia. The positive improvement noted in this study needs to be confirmed in studies with a larger sample size and longer duration.

An In-Vitro Assay Estimating Changes in Melanin Content of Melanoma Cells due to Ultra-Dilute, Potentized Preparations.

INTRODUCTION: The authors had previously conducted an in-vitro study to observe the effect of homeopathic medicines on melanogenesis, demonstrating anti-vitiligo potential by increasing the melanin content in murine B16F10 melanoma cells. A similar experiment was performed using further homeopathic preparations sourced from kojic acid (KA), hydrogen peroxide (H2O2; HP), 6-biopterin (BP), and [Nle4, D-Phe7]-α-melanocyte-stimulating hormone (NLE), some of which are known to induce vitiligo or melano-destruction at physiological dose. MATERIALS AND METHODS: The homeopathic preparations of BP, KA, NLE, and HP were used in 30c potency. Alcohol and potentized alcohol were used as vehicle controls. Prior to starting the main experiment, the viability of B16F10 melanoma cells after treatment with study preparations was assayed. Melanin content (at 48 h and 96 h) and tyrosinase activity in melanocytes were determined. RESULTS: At the end of 48 hours, NLE and HP in 30c potency had a significantly greater melanin content (p = 0.015 and p = 0.039, respectively) compared with controls; BP and KA in 30c potency had no significant effects. No significant changes were seen at the end of 96 hours. KA, NLE, HP, and vehicle controls showed an inhibition of tyrosinase activity. CONCLUSION: The study demonstrated melanogenic effects of two homeopathic preparations. Further research to evaluate the therapeutic efficacy of these medicines is warranted.

An experimental study to evaluate the antiosteoporotic effect of Panchatikta Ghrita in a steroid-induced osteoporosis rat model.

OBJECTIVE: The study was conducted to develop the glucocorticoid-induced osteoporosis (GIO) model in Sprague-Dawley weanling rats using different doses of methylprednisolone (MP) and evaluate the antiosteoporotic effect of a classical ayurvedic formulation, Panchatikta Ghrita (PG), in this model. MATERIALS AND METHODS: Institutional Animal Ethics Committee approval was obtained. Development of model was done by subcutaneous injection of 2 doses of MP (14 and 28 mg/kg/week) for 4 weeks in 21-day old weanlings. Following confirmation of the dose of MP that induced osteoporosis, the antiosteoporotic effect of PG was tested in this model in comparison to a known antiosteoporotic agent, alendronate. Both alendronate (2.9 mg/kg/day) and PG (1.35 g/kg/day) were administered orally 2 weeks after MP - 14 mg/kg/week injection and continued for 4 weeks. Serum and urine calcium and inorganic phosphate were analyzed at weekly intervals. Animals were sacrificed after 6 weeks, and femur bones were processed to measure bone hardness and elasticity and for histological studies. RESULTS: Rats treated with MP - 14 mg/kg/week showed optimum osteoporotic effect with no mortality as compared to MP - 28 mg/kg/week; hence, this dose of MP was used further for the efficacy study. Osteoporotic rats treated with PG 1.35 g/kg showed increase in serum calcium and inorganic phosphate levels, whereas urine calcium and phosphate levels were significantly reduced. A significant decrease in a number of osteoclasts, whereas an increase in bone hardness and elasticity was observed as compared to diseased group demonstrating antiosteoporotic effect of PG. CONCLUSION: PG has an antiosteoporotic effect in GIO rat model.

Immunological & metabolic responses to a therapeutic course of Basti in obesity.

BACKGROUND & OBJECTIVES: Basti (medicated enema) is a popular Ayurvedic intervention recommended for obesity. However, there are no data to show whether any physiological or biochemical changes occur following this treatment. This study was conducted to identify the immunological and metabolic changes in obese individuals after a therapeutic course of Basti. METHODS: Thirty two obese individuals (18 and 60 yr) with a body mass index (BMI) ≥30 kg/m [2] who received a therapeutic course of 16 enemas (Basti) followed by a specific diet and lifestyle regimen for a period of 32 days as their treatment for obesity, were enrolled in the study. Clinical examination, measurement of immune and metabolic markers were done before (S1), immediately after (S2) and 90 days after the completion of therapy (S3). RESULTS: A significant reduction ( P<0.001) in weight, BMI, upper arm and abdominal circumference was seen at S3, along with a decrease in serum interferon (IFN)-γ (P<0.02), interleukin (IL)-6 ( P<0.02) and ferritin (P<0.05) and increase in IgM levels ( P<0.02). Peripheral blood lymphocytes (PBLs) stimulated with anti-CD3 monoclonal antibodies showed significant increase in reactive oxygen species (ROS) generation and calcium flux after Basti. All organ function tests revealed no changes. INTERPRETATION & CONCLUSIONS: Our study documents that a therapeutic course of Basti modulates immune responses by regulating pro-inflammatory cytokines, immunoglobulins and functional properties of T-cells. These changes are associated with a reduction in the body weight which is maintained even after three months of treatment. The study also documents the safety of Basti procedure.

Effects of a food-based intervention on markers of micronutrient status among Indian women of low socio-economic status.

Intakes of micronutrient-rich foods are low among Indian women of reproductive age. We investigated whether consumption of a food-based micronutrient-rich snack increased markers of blood micronutrient concentrations when compared with a control snack. Non-pregnant women (n 222) aged 14-35 years living in a Mumbai slum were randomised to receive a treatment snack (containing green leafy vegetables, dried fruit and whole milk powder), or a control snack containing foods of low micronutrient content such as wheat flour, potato and tapioca. The snacks were consumed under observation 6 d per week for 12 weeks, compliance was recorded, and blood was collected at 0 and 12 weeks. Food-frequency data were collected at both time points. Compliance (defined as the proportion of women who consumed ≥ 3 snacks/week) was >85 % in both groups. We assessed the effects of group allocation on 12-week nutrient concentrations using ANCOVA models with respective 0-week concentrations, BMI, compliance, standard of living, fruit and green leafy vegetable consumption and use of synthetic nutrients as covariates. The treatment snack significantly increased ß-carotene concentrations (treatment effect: 47·1 nmol/l, 95 % CI 6·5, 87·7). There was no effect of group allocation on concentrations of ferritin, retinol, ascorbate, folate or vitamin B12. The present study shows that locally sourced foods can be made into acceptable snacks that may increase serum ß-carotene concentrations among women of reproductive age. However, no increase in circulating concentrations of the other nutrients measured was observed.

Development of an experimental diet model in rats to study hyperlipidemia and insulin resistance, markers for coronary heart disease.

OBJECTIVES: The objective of this study is to develop an experimental model of hyperlipidemia and insulin resistance (IR), markers of coronary heart disease (CHD) using high fat and high sugar (HFHS) diet and to evaluate the efficacy of the model using atorvastatin, a known antihyperlipidemic drug, pioglitazone, a known insulin sensitizer, and Tinospora cordifolia (Tc), an antidiabetic plant. MATERIALS AND METHODS: Following Institutional Animal Ethics Committee permission, the study was conducted in male Wistar rats (200-270 g). The model was developed using a high fat (vanaspati ghee: coconut oil, 3:1) oral diet along with 25% fructose (high sugar) added in drinking water over a period of 6 weeks. Atorvastatin (2.1 mg/kg/day), pioglitazone (2.7 mg/kg/day) and Tc (200 mg/kg/day) were administered 3 weeks after initiation of HFHS diet and continued for another 3 weeks. Parameters assessed were weight, lipid profile, fasting blood glucose, insulin, and gastric emptying. Serum malondialdehyde (MDA) and catalase were assessed as markers of oxidative stress. RESULTS: Administration of HFHS diet demonstrated a significant increase in blood glucose, insulin, total and low density lipoprotein cholesterol and triglycerides with a decrease in high density lipoprotein cholesterol. Treatment with test drugs decreased blood sugar, insulin, lipid parameters, increased gastric emptying rate, decreased MDA levels, and catalase activity when compared to HFHS diet group, confirming the efficacy of the model. Atherogenic index of all the test drugs (0.48, 0.57, and 0.53) was significantly lower as compared to HFHS diet group (1.107). CONCLUSION: This study confirms the development of a diet based cost-effective and time efficient experimental model, which can be used to study two important markers of cardiovascular disease that is, hyperlipidemia and IR and to explore the efficacy of new molecules in CHD.

Do plants mediate their anti-diabetic effects through anti-oxidant and anti-apoptotic actions? an in vitro assay of 3 Indian medicinal plants.

BACKGROUND: Both experimental and clinical studies suggest that oxidative stress plays a major role in the pathogenesis of both types of diabetes mellitus. This oxidative stress leads to ß-cell destruction by apoptosis. Hence exploring agents modulating oxidative stress is an effective strategy in the treatment of both Type I and Type II diabetes. Plants are a major source of anti-oxidants and exert protective effects against oxidative stress in biological systems. Phyllanthus emblica, Curcuma longa and Tinospora cordifolia are three such plants widely used in Ayurveda for their anti-hyperglycemic activity. Additionally their anti-oxidant properties have been scientifically validated in various experimental in vitro and in vivo models. Hence the present in vitro study was planned to assess whether the anti-hyperglycemic effects of the hydro-alcoholic extracts of Phyllanthus emblica (Pe) and Curcuma longa (Cl) and aqueous extract of Tinospora cordifolia (Tc) are mediated through their antioxidant and/or anti-apoptotic property in a streptozotocin induced stress model. METHODS: RINm5F cell line was used as a model of pancreatic ß-cells against stress induced by streptozotocin (2 mM). Non-toxic concentrations of the plant extracts were identified using MTT assay. Lipid peroxidation through MDA release, modulation of apoptosis and insulin release were the variables measured to assess streptozotocin induced damage and protection afforded by the plant extracts. RESULTS: All 3 plants extracts significantly inhibited MDA release from RIN cells indicating protective effect against STZ induced oxidative damage. They also exhibited a dose dependent anti-apoptotic effect as seen by a decrease in the sub G0 population in response to STZ. None of the plant extracts affected insulin secretion from the cells to a great extent. CONCLUSION: The present study thus demonstrated that the protective effect of the selected medicinal plants against oxidative stress induced by STZ in vitro, which was exerted through their anti-oxidant and anti-apoptotic actions.

Insulin sensitizing effect of 3 Indian medicinal plants: an in vitro study.

OBJECTIVE: Measurement of glucose uptake into peripheral tissue is an important mechanism to assess Insulin sensitivity. The present in vitro study was conducted to evaluate the Insulin sensitizing activity of Phyllanthus emblica (Pe), Tinospora cordifolia (Tc) and Curcuma longa (Cl) by assessing glucose uptake activity in a 3T3L1 adipocyte model. MATERIALS AND METHODS: The 3T3 L1 fibroblast cells were differentiated to adipocytes, using a cocktail of insulin, isobutyl-1-methylxanthine and dexamethazone. These adipocytes were initially treated with different concentrations of the selected plants following which 2-deoxy glucose uptake was estimated using a radioactive assay. The effects of plants on glucose uptake both in the presence and absence of insulin was evaluated and compared with pioglitazone, a known insulin sensitizer. RESULTS: Pe and Tc per se significantly stimulated glucose uptake in 3T3-L1 adipocytes in a dose dependent manner with maximal effect at higher concentrations (200 µg/ml). The effect of both Pe and Tc at 200 µg/ml was comparable to insulin and greater than pioglitazone. Cl per se stimulated glucose uptake with maximal effect at 50 µg/ml. However, this effect was lesser as compared to insulin with higher concentrations inhibiting glucose uptake. When combined with insulin, an antagonist effect was observed between Pe, Tc and insulin indicating a possible plant-drug interaction while Cl in combination with insulin showed an increase in the glucose uptake as compared to Cl alone. CONCLUSION: The results suggest that one of the mechanisms for the anti-diabetic effect of Pe, Cl and Tc may be through an insulin sensitizing effect (stimulation of glucose uptake into adipocytes). Further studies using other target sites viz. skeletal muscle and hepatocytes models and in an insulin resistant state would help substantiate this conclusion.

Polysaccharide enriched immunomodulatory fractions from Tinospora cordifolia (Willd) miers ax hook. f. & Thoms.

Tinospora cordifolia is used in Ayurveda as "Rasayanas" to improve the immune system and the body resistance against infections. Polysaccharides are the main constituents which are considered to be responsible for immune enhancement. In this study, immunomodulatory activity of three polysaccharide enriched fractions was evaluated using the polymorphonuclear leukocyte function test. Sugar composition was determined by GC-MS analysis of the derivatised fractions. The active polysaccharide fractions mainly constitute glucose, fructose and arabinose as monomer units.