Thalamic superoxide and peroxide handling capacity (SPHC): An experimental study with aluminum, ethanol and tocopherol in rats.

Superoxide and peroxide handling capacity (SPHC) is an important determinant of oxidative stress. Neurotoxic impacts of aluminum are associated with oxidant imbalance. Here, we studied the influence of aluminum on oxidative stress parameters, antioxidative enzymes and SPHC of thalamic area on pro-oxidant (ethanol) and antioxidant (α-tocopherol) exposure. Two sets of male Wistar rats were divided into 8 groups (6 each) and exposed to aluminum (10 mg/Kg body wt.), ethanol (0.6 g/Kg body wt.) and α-tocopherol (5 IU/day) for 4 wk, each having respective control group. Levels of reduced glutathione (GSH), lipid peroxidation (TBARS) along with activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione reductase (GR) of thalamic area were estimated for each group. Glutathione-independent superoxide peroxide handling capacity (GI-SPHC) and glutathione-dependent superoxide peroxide handling capacity (GD-SPHC) were calculated from the GPx, CAT and SOD values. Concomitant exposure to aluminum and ethanol demonstrated significant increase in SOD activity and significant decrease in GPx activity compared to the control group, while lone aluminum-exposed rats showed raised GR activity, without alterations in GPx and SOD activities. However, significant reduction of both GI- and GD- SPHC were found in ethanol-exposed groups. α-Tocopherol supplementation could resist most of the alterations. In addition, current antioxidant exposure reduced the inherent GD-SPHC, and thus, made thalamic area more vulnerable to oxidant threat. The present study corroborates the thalamic susceptibility to aluminum-augmented oxidant imbalance and suggests cautious use of antioxidant supplementation against neurodegenerative disorders.

Drug development from natural resource: a systematic approach.

Modern research in drug discovery from medicinal plants involves a multidimensional approach combining botanical, phytochemical, biochemical combinatorial chemistry and bioassay-guided fractionation approaches. Natural sources continue to provide an alternative as pharmacological leads against various devastating diseases such as diabetes, CVD, cancer etc. Nowadays, there is enormous requirement of safe and effective drugs in the world. This has prompted scientists to revert back towards natural resources as a potential source of therapeutics for treatment and management of such chronic and fatal diseases. However, there are certain serious challenges and limitations in this field including scale up and commercialization of active compounds which allow only one in thousand lead molecules to be developed as drug. A systematic and scientific approach is an essential requirement for drug development from natural resource. This mini review provides an overview of the methods involved in natural product research starting from crude plant extract to bioactive pharmacological lead. Moreover, it also discusses the limitations of working concerning the bioactivity of medicinal plants.

Protective effects of Eugenia jambolana extract versus N-acetyl cysteine against cisplatin-induced damage in rat testis.

To assess the protective effects of Eugenia jambolana extract (EJE) or N-acetyl cysteine (NAC) on testis, cisplatin (CIS, 5 mg kg(-1) bw, single dose) was administered either alone or along with EJE (25 mg kg(-1) bw, alternate day) or NAC (150 mg kg(-1) bw, Day 1 and 4) for 7 days. Significant alterations in serum LH, FSH and testosterone were observed in CIS group which were effectively modulated by EJE or NAC supplementation. Upregulation of 3ß-HSD gene indicated the rise in functional Leydig cells. This was further confirmed from the identical improvement in hCG-stimulated testosterone production in isolated Leydig cells. Reduction in oxidative stress was associated with restoration of total antioxidant capacity and glutathione levels, and activation of antioxidant enzymes, SOD, catalase, glutathione s-transferase (GST) and glutathione reductase (GR). CIS-induced apoptosis of germ and Leydig cells was contained by both NAC and EJE intervention by effective modulation of apoptotic markers in the extrinsic, intrinsic and other pathways of metazoan apoptosis. Taken together, the study findings establish the potential of EJE as a therapeutically better antioxidant than NAC for use in curtailing the adverse effects of anticancer drugs on testicular function.

Cytoprotective effects of fruit pulp of Eugenia jambolana on H 2 O 2 -induced oxidative stress and apoptosis in rat Leydig cells in vitro.

This study was undertaken to investigate the cytoprotective effect of the fruit pulp of Eugenia jambolana (50-250 µg ml(-1) ) against the damage induced by H 2 O 2 (100 µm) exposure to Leydig cells in vitro. Cell survival with extract was found comparable to similar effects by N-acetyl-l-cysteine. H 2 O 2 -induced rise in thiobarbituric acid reactive substance formation and decline in the activity and expression of antioxidant enzymes like superoxide dismutase, catalase and glutathione-s-transferase were effectively checked. Cellular glutathione and total antioxidant capacity demonstrated significant improvement. The increase in expression of inducible nitric oxide (NO) synthase leading to NO production was successfully countered. Co-treatment of the extract helped in the down-regulation of caspase-3 and poly-ADP-ribose polymerase resulting in a significant reduction in Leydig cell apoptosis induced by H 2 O 2 . Upstream marker proteins of extrinsic (caspase-8, Fas, FasL) and intrinsic (caspase-9) pathway of metazoan apoptosis were identically down-regulated. The Bcl-2 family of proteins, though, remained unaffected. The extract also positively modulated the other marker proteins like c-Jun NH 2 -terminal kinase, p38, Akt, nuclear factor-κB, c-Fos, cellular FLICE-inhibitory protein, cyclooxygenase-2 and p53. Taken together, the above-mentioned findings establish the anti-oxidative and anti-apoptotic potency of the extract that ameliorates the H 2 O 2 -induced adverse effects on rat Leydig cells in vitro.

Aluminum and ethanol induce alterations in superoxide and peroxide handling capacity (SPHC) in frontal and temporal cortex.

Aluminum is an omnipresent neurotoxicant and has been associated with several neuropathological disorders. Cerebrum and cerebellum have been shown to face augmented oxidative stress when animals are exposed to aluminum and high doses of ethanol. To establish the link between oxidative stress and neurobehavioral alterations, the present study was conducted to determine the extent of oxidative stress in low levels of pro-oxidant (ethanol exposure) status of the functionally discrete regions of the cerebrum. Male Wistar rats were exposed to aluminum (10 mg/kg body wt) and ethanol (0.2-0.6 g/kg body wt) for 4 weeks. Spontaneous motor activity (SMA) and Rota-Rod performances (RRP) were recorded weekly during the period of exposure. At the end of 4th week, oxidative stress parameters were determined from the homogenized cerebral tissue. GSH-independent superoxide peroxide handling capacity (GI-SPHC) and GSH-dependent superoxide peroxide handling capacity (GD-SPHC) were determined for FC and TC upon exposure to ethanol in the absence and presence of aluminum exposure. Aluminum was found to augment the oxidative stress at higher doses (0.6 g Ethanol/kg body wt) of ethanol, particularly in FC. The SPHC of FC was also found to be compromised significantly in aluminum-ethanol co-exposed animals. It was concluded that even though the manifestation of oxidative stress was not observed as revealed by assaying the widely used oxidative stress biochemical markers (indices), aluminum and ethanol (low doses) exposure induced alterations in the handling capacity of oxidant imbalance that could be recognized by studying the SPHC of FC. Comparison of GD-SPHC and GI-SPHC offered a possible mechanism of compromised SPHC in FC. This observation is likely to offer insights into the mechanism of association between aluminium exposure and behavioral changes in neurodegenerative disorders towards therapeutic strategies for these disorders.

Hypoglycaemic and hypolipidemic effect of ethanolic extract of seeds of Eugenia jambolana in alloxan-induced diabetic rabbits.

The hypoglycaemic and hypolipidemic effect of ethanolic extract obtained from seeds of E. jambolana was investigated in alloxan-induced diabetic rabbits. Hypoglycaemic activity was assessed by reduction in fasting blood glucose (FBG) at 90min and also fall in peak blood glucose during glucose tolerance test (GTT) in sub-diabetic and mild diabetic (MD) rabbits, but in severe diabetic (SD) rabbits by reduction in FBG at 90min. Ethanolic extract (100mg/kg body weight) when given orally to sub-diabetic (AR) for 1 day, MD for 7 days and SD for 15 days showed significant fall in FBG at 90min (12% AR, 18.9% MD and 29% SD) and also produced 16.9% fall in peak blood glucose in AR and 21% in MD rabbits during GTT. When administered daily for 15 days to MD and SD rabbits, significant fall in FBG (41.3% MD, 31.6% SD) and glycosylated haemoglobin (GHb) levels (23.3% MD, 26.6% SD) were observed, while serum insulin level showed significant increase (32.8% MD, 26.9% SD). Liver and muscle glycogen content also increased. The ethanolic extract of seeds also exhibited significant hypolipidemic effect as evident from fall in total serum cholesterol (TC)/high density lipoprotein cholesterol (HDL-c) ratio, serum low density lipoprotein cholesterol (LDL-c) levels and decreased activity of HMG-CoA reductase. The histopathological studies of liver, pancreas and aorta in alcoholic extract treated diabetic groups revealed almost normal appearance.

Effect of Yoga asanas on nerve conduction in type 2 diabetes.

Twenty Type 2 diabetic subjects between the age group of 30-60 years were studied to see the effect of 40 days of Yoga asanas on the nerve conduction velocity. The duration of diabetes ranged from 0-10 years. Subject suffering from cardiac, renal and proliferative retinal complications were excluded from the study Yoga asanas included Suryanamskar. Tadasan, Konasan, Padmasan Pranayam, Paschimottansan Ardhmatsyendrasan, Shavasan, Pavanmukthasan, Sarpasan and Shavasan. Subjects were called to the cardio-respiratory laboratory in the morning time and were given training by the Yoga expert. The Yoga exercises were performed for 30-40 minutes every day for 40 days in the above sequence. The subjects were prescribed certain medicines and diet. The basal blood glucose, nerve conduction velocity of the median nerve was measured and repeated after 40 days of Yogic regime. Another group of 20 Type 2 diabetes subjects of comparable age and severity, called the control group, were kept on prescribed medication and light physical exercises like walking. Their basal & post 40 days parameters were recorded for comparison. Right hand and left hand median nerve conduction velocity increased from 52.81 +/- 1.1 m/sec to 53.87 +/- 1.1 m/sec and 52.46 +/- 1.0 to 54.75 +/- 1/1 m/sec respectively. Control group nerve function parameters deteriorated over the period of study, indicating that diabetes is a slowly progressive disease involving the nerves. Yoga asanas have a beneficial effect on glycaemic control and improve nerve function in mild to moderate Type 2 diabetes with sub-clinical neuropathy.

Study of yoga asanas in assessment of pulmonary function in NIDDM patients.

Certain yoga asanas if practiced regularly are known to have beneficial effects on human body. These yoga practices might be interacting with various, somato-neuro-endocrine mechanisms to have therapeutic effects. The present study done in twenty four NIDDM patients of 30 to 60 year old, provides metabolic and clinical evidence of improvement in glycaemic control and pulmonary functions. These middle-aged subjects were type II diabetics on antihyperglycaemic and dietary regimen. Their baseline fasting and postprandial blood glucose and glycosylated Hb were monitored along with pulmonary function studies. The expert gave these patients training in yoga asanas and were pursed 30-40 min/day for 40 days under guidance. These asanas consisted of 13 well known postures, done in a sequence. After 40 days of yoga asanas regimen, the parameters were repeated. The results indicate that there was significant decrease in fasting blood glucose levels (basal 190.08 +/- 90.8 in mg/dl to 141.5 +/- 79.8 in mg/dl). The postprandial blood glucose levels also decreased (276.54 +/- 101.0 in mg/dl to 201.75 +/- 104.1 in mg/dl), glycosylated hemoglobin showed a decrease (9.03 +/- 1.4% to 7.83 +/- 2.6%). The FEV1, FVC, PEFR, MVV increased significantly (1.81 +/- 0.4 lt to 2.08 +/- 0.4 lt, 2.20 +/- 0.6 lt to 2.37 +/- 0.5 lt, 3.30 +/- 1.0 lt/s to 4.43 +/- 1.4 lt/s and 64.59 +/- 25.7 lt min to 76.28 +/- 28.1 lt/min respectively). FEV1/FVC% improved (85 +/- 0.2% to 89 +/- 0.1%). These findings suggest that better glycaemic control and pulmonary functions can be obtained in NIDDM cases with yoga asanas and pranayama. The exact mechanism as to how these postures and controlled breathing, interact with somato-neuro-endocrine mechanism affecting metabolic and pulmonary functions remains to be worked out.

Anti-inflammatory and analgesic effect of herbal preparation: septilin.

Septilin a proprietary herbal preparation has been reported to have wound healing and immunomodulatory properties. The present study was, therefore, designed to ascertain the anti-inflammatory and analgesic properties of septilin in rats and mice by oral route. Anti-inflammatory activity was assessed by using acute (carrageenan-induced paw oedema), sub-acute (cotton pellet granuloma) and chronic (Freund's adjuvant induced-arthritis) models of inflammation. Administration of septilin in a dose of 500 mg/kg orally produced significant anti-inflammatory effect in all experimental models of inflammation. However, the anti-inflammatory effect was more in chronic model when compared to reference standard drug (Prednisolone). Septilin (500 mg/kg) produced significant analgesic effect on tall flick test. However, the effect was less when compared to morphine (5 mg/kg) but when septilin was combined with subanalgesic dose of morphine (2.5 mg/kg) the analgesic effect was more than the either drug given alone. On GAA induced writhing test septilin produced significant analgesic effect at a dose of 750 mg/kg which was comparable to diclofenac (50 mg/kg). However, subanalgesic dose of septilin (500 mg/kg) when combined with diclofenac (25 mg/kg), analgesic effect was enhanced significantly.

A preliminary report on the role of yoga asanas on oxidative stress in non-insulin dependent diabetes mellitus.

Nineteen subjects of non-insulin dependent diabetes mellitus (NIDDM) between the age group of 30-60 yrs were studied to see the effect of specific yoga asanas on fasting and postprandial blood glucose (FBG, PPG), serum malondialdehyde (MDA) and glycosylated hemoglobin (HbA(1)) in addition to drug treatment and diet control. The duration of diabetes ranged from 1-10 years. Patients with renal, cardiac and proliferative retinal diseases were excluded from the study. The same patients served as their own control. Subjects were called in the morning to the cardio-respiratory laboratory and were given training by a yoga expert. Yoga asanas included Suryanamskar, Tadasan, TriKonasan, Padmasan, Pranayam, Paschimottanasan, Ardhmatsyendrasan, Pavanmukthasan, Sarpasan and Shavasan. The asanas were done every day for 40 days for 30-40 min. FBG, PPG, serum MDA and HbA(1) were estimated before and after 40 days of yoga asanas regimen. Significant reduction was seen in FBG from 220 mg/dl to 162 mg/dl, PPG from 311 mg/dl to 255 mg/dl, MDA from 6 nmol/l to 3 nmol/l and HbA(1), from 8.8% to 6.4%. Subjects felt better and were relieved of their stresses and had an improvement in their day to day performance. The decrease was statistically significant (p<0.0001 for FBG and PPG, p<0.001 for MDA and for HbA(1)).

Medicinal plants for treatment of diabetes mellitus.

Many plants have been used for the treatment of diabetes mellitus in Indian system of medicine and in other ancient systems of the world. Out of these only a few have been evaluated as per modern system of medicine. From many such plants only extracts have been prepared and their usefulness evaluated in experimental diabetes in animals. In some plants likeAllium cepa, Allium sativum, Ficus bengalensis, Gymnema sylvestre, Pterocarpus marsupium etc. active hypoglycemic principles have been isolated and their mechanism of action studied. Most of them seem to act directly on pancreas (pancreatic effect) and stimulate insulin level in blood. Some have extra pancreatic effect also by acting directly on tissues like liver, muscle etc. and alter favourably the activities of the regulatory enzymes of glycolysis, gluconeogenesis and other pathways. Since the plant products have less side effects, they have the potential as good hypoglycemic drugs. They may also provide clues for the development of new and better oral drugs for diabetes.

Alternative approach and evaluation of three potency assay methods for testing adsorbed tetanus toxoid.

Thirteen batches of adsorbed Tetanus Toxoid (TT) from different manufacturers were tested for potency by three different methods viz: (i) An Antibody Induction Method (AIM) developed in mice: (ii) WHO lethal challenge in mice; and (iii) Conventional Antibody Induction (I.P). Method in guinea pigs. The potency results obtained in AIM, by serological evaluation of immunized mice were found identical and correlated significantly with those obtained by WHO recommended lethal challenge test in mice. The potency data obtained in the present study was found comparable with other studies. An AIM in mice thus offers an alternative to lethal challenge tests and can replace guinea pig model. Out of 107 serum samples obtained from immunized guinea pigs in the conventional antibody induction method, 90% samples contained more than 4 units of tetanus antitoxin per ml. End point titres of 42 serum samples belonging to 5 batches of TT also showed much higher tetanus antitoxin content when determined by TN test. The potency data obtained thus suggest revision of the minimum requirement in Indian Pharmacopoeia which is too low and which may be increased as indicated by the present study.

Quantitation of tetanus and diphtheria antitoxins in mouse sera by indirect haemagglutination.

Serum samples obtained from 75 groups of mice immunized with various doses of adsorbed tetanus vaccine, adsorbed diphtheria-tetanus vaccine and adsorbed diphtheria-tetanus-pertussis vaccine were titrated for tetanus antitoxin content by an in-vitro indirect haemagglutination (IHA) and by toxin neutralization test (TN) in mice. From these serum samples of 49 groups of mice which were immunized with combined vaccine containing diphtheria toxoid were titrated for their diphtheria antitoxin content by IHA and by i.d. toxin neutralization test (TN) in guinea pigs. Good correlations were found between the estimates obtained by in-vitro IHA and in vivo TN tests in both tetanus and diphtheria antitoxin titrations. The minimum level of tetanus or diphtheria antitoxin detectable by IHA was 0.00039 IU/ml. It is concluded that IHA is a simple, sensitive and reproducible alternative test which can replace the animal TN tests for the estimation of tetanus and diphtheria antitoxins and could reliably be used in the potency assay of tetanus and diphtheria toxoids of combined vaccines based on antibody induction in mice.

Biochemical studies on combined effects of garlic (Allium sativum Linn) and ginger (Zingiber officinale Rosc) in albino rats.

Adult male rats of wistar strain were fed diets containing (w/w) 2% garlic (group II), 0.5% ginger (group III) and a combination of garlic plus ginger (group IV) for 4 weeks to study their effects on serum biochemical parameters. A significant increase in body weight was observed in all groups except that fed ginger (group III). A significant decrease in blood glucose, serum total cholesterol and serum alkaline phosphatase were found in all groups, whereas serum triglycerides were decreased significantly only in group IV. Serum HDL-cholesterol was significantly increased only in groups III and IV. However, HDL-cholesterol, VLDL-cholesterol and atherogenic index were significantly decreased in animals fed with combination of the two, compared to garlic/ginger alone. Hence, a combination of garlic and ginger is much more effective in reducing blood glucose and serum lipids.

Effect of herbal preparation on immune response of immunosuppressed mice.

The present study was undertaken to study the effect of a herbal formulation (Septilin) as immunomodulator, on immune response in mice. The study of this formulation in respect to humoral and cell mediated immune response has suggested that, oral administration of Septilin (500 mg/kg) alone or in combination with an immuno-suppressive drug (prednisolone 4 mg/kg), enhances both primary and secondary immune response, in mice immunized with sleep red blood cells (SRBC).