Influence of Prooxidant Exposure on Aluminum-induced Alterations in Open-field Behavioral Study in Rats.

Neurotoxic role of aluminum is being implicated in the neurodegenerative changes and neurobehavioral pathology. Prooxidant (ethanol) exposures were found to augment the neurochemical alterations in different brain regions. Current study was aimed to follow up aluminum-induced alterations in open-field behavior of rats in absence and presence of varied doses of ethanol (prooxidant) exposure for four weeks. Male Wistar rats were exposed to oral aluminum chloride gavage (Al+ animals; 10 mg/Kg bw) and concomitantly treated with ethanol (0.2, 0.4 and 0.6 g/Kg bw) daily. Open-field behavior of rats were recorded weekly and processed for ambulatory, thigmotaxic and spatial behaviors; and compared with controls – Al0 (without aluminum exposure) and Et-0 (without ethanol exposure). Lone aluminum exposure was found to alter the quadrant preferences only at the end of 4th week of treatment. However, differences in quadrant preferences between Al0 and Al+ animals were observed at the end of 1st week itself in presence of prooxidant exposures. Significant influences of aluminum was seldom during the initial weeks and were restricted to right angle turn and wall climbing behaviors, while many parameters were significantly influenced by ethanol exposures at the end of third week. By the end of 4th week, ambulation, thigmotaxis and quadrant preferences were found to be significantly influenced by either aluminum or ethanol exposures and/or their interactions. The study clearly indicates that certain aspect of neurobehavioral toxicity of aluminum can be aggravated by concomitant presence of prooxidant dominance. Present investigation accentuates the role of aluminum toxicity in behavioral neuropathology and revealed that there are at least two specific mechanisms of aluminum-induced neurobehavioral alterations – one is oxidative stress dependent while the other is not.

Waardenburg Syndrome Presenting With Constipation Since Birth.

Background: Shah-Waardenburg syndrome is Waardenburg syndrome associated with Hirschsprung’s disease. Case characteristics: A 10-day-old full-term male neonate of Waardenburg syndrome presented with constipation since birth along with features of small bowel obstruction. Observation: Exploratory laparotomy revealed distended proximal jejunal and ileal loops along with microcolon; an ileostomy was performed. Outcome: Postoperatively patient developed sepsis and died. Histopathology confirmed total colonic aganglionosis. Message: Suspect familial Shah-Waardenburg syndrome in a neonate of Waardenburg syndrome presenting with constipation since birth or intestinal obstruction.

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.

Coronary risk variables in young asymptomatic smokers.

BACKGROUND AND OBJECTIVES: Smoking plays a dominant role in premature atherosclerosis particularly among males in South Asian countries. It initiates and promotes atherosclerosis by altering cardiac haemodynamics, causing dyslipidaemia and producing oxidative damage. Not much information is available from our country. We therefore undertook this study to see the effect of smoking on electrocardiogram (ECG), blood pressure, lipids, apolipoprotein B level and free radical activity in young asymptomatic male smokers. METHODS: The study included 100 consecutive male subjects (50 smokers and 50 non smokers) aged 30-40 yr. Smoking profile, detailed cardiovascular assessment including ECG and lipid profile were evaluated in each subject. RESULTS: Of the 50 smokers, 22 (44%) had grade I hypertension as against 5 of 50 non smokers. Sinus tachycardia (10%) and P-pulmonale (8%) were the only notable ECG abnormalities. Dyslipidaemia was detected in 92 per cent smokers and 48 per cent non smokers (P<0.001). Total serum cholesterol, low density lipoprotein (LDL)-cholesterol, triglycerides and apolipoprotein B levels were significantly higher (P<0.001) in smokers compared to non smokers. LDL-cholesterol was > or =135 mg/dl in 94 per cent dyslipidaemic smokers. However, no significant difference was found in high density lipoprotein (HDL)-cholesterol. Smokers had significantly higher serum malondialdehyde levels (P<0.001) and low superoxide dismutase (P<0.001) compared to non smokers. INTERPRETATION AND CONCLUSION: Our data indicate that young asymptomatic male smokers tend to have hypertension, dyslipidaemia and increased production of free oxygen radicals, perhaps by attenuation of oxidative stress by cigarette smoking. This makes them prone for premature coronary artery disease. However, the findings need to be confirmed on a larger sample.

Ectopic pancreas as a cause of gastric outlet obstruction in an infant.

We report a one-month-old male child who presented with clinical and radiological features of gastric outlet obstruction. Surgical exploration showed presence of a prepyloric mass; histological examination of the resected specimen confirmed presence of ectopic pancreatic tissue. The child is well 8 months later.

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.

Studies on oxidative stress, serum iron and iron binding capacity in subjects prone to the risk of coronary artery disease.

Lipid peroxidation in vitro and in vivo has been postulated to be involved in the development of atherosclerosis. It is also known that free iron catalyses the lipid peroxidation. Therefore, we assessed the status of oxidative stress in smokers, hypertensives and non-insulin dependent subjects, who were prone to coronary artery disease. In addition, superoxide dismutase levels and iron binding capacity were also measured to know their antioxidant defences. One hundred seventy-five consecutive subjects below 60 years of age were examined; they were then divided into three groups: one with coronary artery disease, another without coronary artery disease and a healthy control group. The patients having either of the one risk factors for coronary artery disease i.e. smoking, hypertension and/or diabetes were studied. Serum lipid peroxides, superoxide dismutase, serum iron and iron binding capacity were estimated. Oxidative stress was highest in smokers with coronary artery disease (3.11+/-0.79 mmol/ml) as compared to hypertensives (2.69+/-0.20 mmol/nl) and non-insulin dependent diabetics (2.78+/-0.19 mmol/ml). Superoxide dismutase activity was also significantly decreased (p<0.001) in smokers with coronary artery disease as compared to hypertensives and non-insulin dependent diabetes mellitus. Final step of stepwise logistic regression based on malondialdehyde and superoxide dismutase correctly predicted coronary artery disease status in 90 percent smokers. Serum iron and total iron binding capacity were not significantly different in risk prone subjects. However, among all risk prone subjects, smokers with coronary artery disease showed highest serum iron levels and decreased iron binding capacity.

An antibody induction method in mice for the potency assays of diphtheria and tetanus components in combined vaccines.

Eleven batches of Adsorbed Diphtheria-Tetanus (DT) vaccines and thirteen batches of Adsorbed Diphtheria-Pertussis-Tetanus (DTP) vaccines were tested for the potency of diphtheria and tetanus components by an Antibody Induction Method (AIM) developed in mice. The potency results obtained were found comparable and did not show any statistically significant difference with those obtained by WHO recommended lethal challenge tests for diphtheria in guinea pigs and for tetanus in mice. AIM in mice is more economical as both diphtheria and tetanus components of combined vaccine can be tested in the same experiment and the procedure also eliminates the use of guinea pigs required in the lethal challenge/conventional tests. The data obtained while testing tetanus component by the conventional antibody induction (IP) method in guinea pigs suggests that minimum requirements laid down in i.p. is too low which may be fixed as at least 3 out of 9 guinea pig sera and should contain > or = 4 units of tetanus antitoxin per ml.

"Changing trends (clinico-biochemical) in gall-bladder stone disease"--an observation.

Present study has been undertaken to know the causative factors responsible for change in trend of gall-stone disease from middle aged, fertile, fat females to young asthenic females in twenties. Our findings reveal high incidence of gall stone formation in non-obese young females. Average fat consumption in non-obese patients was less (17%) than that of obese (26%). However, use of oral contraceptives was high in non-obese females and maximum users were in young age group while in obese in middle age group. Bilirubin content in gall bladder stones of non-obese was significantly more than that of obese (p < 0.01) whereas cholesterol content in gall bladder stones of obese was significantly high when compared to non-obese subjects. Analysis of bile showed significant increase in bilirubin and calcium level of non-obese when compared to control and obese subjects whereas phosphorus levels were significantly decreased in the bile of non obese subjects. These findings suggest that in non-obese females less intake of fat, early use of oral contraceptives, higher contents of bilirubin and calcium and low content of phosphorus in bile may be responsible for gall stone formation.

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).

Transplacental immunity and waning of maternal antibody in measles.

Since transplacental immunity and waning of maternally derived measles specific antibodies play an important role in determining the optimum age for vaccination of infants against measles, a study was carried out in which 150 paired samples and 581 infant serum samples were tested for measles specific antibodies. Out of these paired samples, 132 pairs showed measles antibodies in both mother and cord. HAI antibody was absent in 3 paired samples whereas, 5 mothers could not pass on the antibodies in the cord samples. In the remaining 10 serum samples only cord blood showed the presence of antibodies without the detectable level of antibodies in mother. Statistically no significant difference between the mother and cord blood titers was observed by applying the student 't' test for comparison of the mean (t = 0.01). Analysis of 581 infant serum samples for prevalence of maternal antibodies indicated that 83% of the samples at the age of 3 months or below had measles antibodies but with the increase in age there was tremendous loss with only 19-20% at the age of 6-7 months. After 7 months the percentage of infants which had antibody varied from 11-13%. There was negative correlation between age and seropositivity (r = -0.72) which was highly significant (p < 0.05).

Evaluation of potency of inactivated cholera vaccine by mouse protection assay & antibody induction method.

Twenty one batches of whole cell inactivated cholera vaccine manufactured at Central Research Institute, Kasauli were evaluated for potency by mouse protection assay (MPA) and antibody induction method. In the antibody induction method the sera of immunized mice were screened for the presence of antibodies against Vibrio cholerae by microagglutination (MA) test and IgG ELISA. The number of organisms estimated by MPA were correlated with agglutinating and neutralizing antibodies against individual serotypes by MA and ELISA respectively. Correlation coefficient(r) of 0.692 and 0.815 were observed for the titres evaluated by MA and ELISA when compared with standard MPA method for the serotype Ogawa. Similarly r values of 0.925 and 0.849 were observed for titres evaluated by MA and ELISA when compared with standard MPA method for the serotype Inaba. Antibody induction method can be as an alternative method for determining the potency of inactivated cholera vaccine.

Comparative stabilities of glutaraldehyde & heat inactivated pertussis vaccine components of adsorbed DPT vaccine with different preservatives.

The stability of pertussis component (glutaraldehyde or heat inactivated pertussis vaccine) of the adsorbed diphtheria-pertussis-tetanus (DPT) vaccine preserved in thiomersal or benzethonium chloride was studied at 4-8 degrees C and 35 degrees C for 30 days. The potency of pertussis component of adsorbed DPT vaccine preserved with benzethonium chloride was lower than that preserved with thiomersal. After the initial loss of potency of pertussis component in the benzethonium chloride during blending, the stability of potency of pertussis component at 4-8 degrees C and 35 degrees C for 30 days was similar for vaccines preserved with either benzethonium chloride or thiomersal. The stability of both types of pertussis components inactivated with glutaraldehyde or heat was also similar at both the temperatures for 30 days.