Production Purification and Characterization of Extracellular Anti-Leukaemic L-Asparaginase from Isolated Bacillus Subtilis Using Solid State Fermentation.

Bacterial L-asparaginase has been widely used as therapeutic agent in treatment of various lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. The present work deals with production and purification of extracellular L-asparaginase from soil isolates using solid state fermentation. The isolate was characterized by big chemical test and identified as Bacillus subtilis. The enzyme production was carried out by solid state fermentation comparing the results with submerged fermentation. The enzyme was purified to near homogeneity by ammonium sulphate precipitation, dialysis, followed by gel filtration on Sephadex G-100 column, CM Sephadex C-50 and SDS-PAGE. The enzyme was purified at 110.2 folds and showed a final specific activity of 1785.7 IU/mg with 26.5% yield. SDS-PAGE of the purified enzyme revealed an apparent molecular weight of 109 kDa. The purified enzyme showed maximum activity at pH 9 when it was incubated at 50°C for 35 min. The enzyme was activated by Mg+2 and strongly inhibited by EDTA.

Purification and characterization of an extracellular alkaline protease produced from an isolated bacillus subtilis.

This paper describes the studies on the purification and partial characterization of serine alkaline protease produced through submerged fermentation process from a locally isolated Bacillus subtilis. This strain, grown in a highly alkaline medium (pH 10), produces an extracellular proteolytic enzyme. The alkaline protease was purified in a simple two-step procedure involving ammonium sulphate precipitation and gel filtration. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the purified alkaline protease indicated an estimated molecular mass of 30KDa. It was more active in the range of 20-60ºC and had an optimum activity at 55ºC with optimum pH of 10.5. Characterization of the protease showed that it required certain cations such as Mg++, Mn++ and Ca++ for maximal activity. The serine nature of the alkaline protease was confirmed by PMSF inhibition. The temperature and pH stability of this Alkaline Protease from Bacillus Subtilismakes it potentially useful forindustrial applications.

Production Optimization of Extracellular L-Asparaginase Through Solid- State Fermentation By Isolated Bacillus Subtilis.

L-asparaginase has been used as anti-tumor agent for the treatment of acute lymphoblastic leukemia and food processing aid to reduce the formation of cancer causing acrylamide. Extracellular Lasparaginase production was optimized through solid state fermentation using ground nut cake by isolated Bacillus subtilis. which was not reported in literature.Optimum production of L-asparaginase enzyme (18.4U/ml) was obtained after 48h of incubation at 370C moisture content of 70% and at pH 7.

Scleredema of Buschke: a rare post-streptococcal complication.

A 19-year male presented with acute onset, gradually progressive symmetric indurations involving the skin over the face, neck, shoulders and upper part of chest following an upper respiratory tract infection. Detailed history and examination did not reveal evidence of Raynauds' phenomenon, nail changes, digital ulcers, pigmentation or any systemic involvement. Autoantibodies for systemic sclerosis were absent. Histopathology of skin biopsy documented scleredema. Antistreptolysin O (ASO) titer was elevated. We diagnosed a case of Scleredema adultorum of Buschke following a streptococcal throat infection. We report this case to highlight the importance of clinically differentiating this relatively benign, self-limiting disorder from systemic sclerosis.

Correlation of lipid profile and carotid artery plaque as detected by Doppler ultrasound in ischaemic stroke patients--A hospital-based study.

Stroke is one of the leading causes of morbidity and mortality all over the world. Carotid plaque formation and intima media thickness can be a predictor of ischaemic stroke. In this regard studies from our country, are few and far between. This is a small hospital-based study to look in to this matter. We have assessed the intima media thickness of the common carotid as well as the internal and external carotid arteries by the ultrasound method. The lipid profiles were estimated and correlated with the intima media thickness. Results indicate that in the common as well as in the internal and external carotid arteries, the intima media thickness is a good predictor of ischaemic stroke. This thickness is also well correlated with the lipid levels in blood. Hence this non-invasive method can be used successfully to identify the high risk patients, prone to develop stroke.

Hepatopulmonory syndrome--an uncommon and severe complication of certain liver diseases.

Hepatopulmonary syndrome (HPS) is charaterised by arterial hypoxaemia found mainly in association with chronic liver disease (most commonly cirrhosis of liver) but also rarely with acute liver diseases like fulminant hepatic failure or Budd-Chiari syndrome. The purpose of this article is to present an uncommon but grave complication mostly associated with cirrhosis of liver and to differentiate this entry from cyanotic heart diseases or certain lung conditions. Investigations include history and physical examination, certain biochemical tests, upper gastro-intestinal (GI) endoscopy, abdominal ultrasonography, liver biopsy and venacavography in appropriate cases. Finally, the suspected cases underwent arterial blood gas (ABG) analysis and contrast enhanced echocardiography (CEE) for confirmation of the diagnosis of HPS. Of the 123 cases of cirrhosis of liver, three cases of HPS (2.4%) were found-all of them being males. Also one male patient with inferior vena cava (IVC) obstruction amongst other causes presenting with HPS was encountered. As of now, no medical treatment has been proved to be useful and liver transplant remains the only hope for this disorder.

Glycaemic consequences of HIV and highly active antiretroviral therapy: a pilot study and review of literature.

The metabolic consequences of HIV and AIDS are accentuated in the setting of highly active antiretroviral therapy. Peripheral lipodystrophy, central adiposity, hyperlipidaemia, insulin resistance and diabetes mellitus are frequent associations of protease inhibitor containing highly active antiretroviral therapy regimens. Ninety patients aged 25-50 years (males 52, females 38), seropositive for HIV 1 and 2 or both were selected to see the glycaemic profiles in asymptomatic early HIV disease with CD4 counts > 100/microl and to compare this with the glycaemic profile of (a) advanced HIV disease (CD4 counts < 200/microl), not on highly active antiretroviral therapy and (b) advanced HIV disease (CD4 counts < 200/microl), on uninterrupted non-protease inhibitor highly active antiretroviral therapy > 6 months. All the patients were grouped into 3: (1) Group A: CD4 counts > 500/microl (n=37), highly active antiretroviral therapy naive, (2) group B: CD4 counts < 200/microl (n=21), not on highly active antiretroviral therapy, and (3) group C: CD4 counts < 200/microl, receiving uninterrupted non-protease inhibitor based highly active antiretroviral therapy for > 6 months (n=32). The fasting blood glucose, glycosylated Hb (HbA1c) levels, were measured in all the patients in 3 groups and significance of difference between means was calculated among various groups. Body weight and waist-hip ratio were also measured. The results were analysed and compared with other studies.

Comparison of ondansetron, dexamethasone, ondansetron plus dexamethasone and placebo in the prevention of nausea and vomiting after laparoscopic tubal ligation.

Laparoscopic tubal ligation is associated with an appreciably high rate of postoperative nausea and vomiting. This study was designed to compare the effectiveness of ondansetron, dexamethasone, ondansetron plus dexamethasone or placebo in the prevention of postoperative nausea and vomiting in patients after laparoscopic tubal ligation. In a prospective, randomised, double blind placebo controlled trial, 160 ASA I-II females received one of four regimens; ondansetron 4 mg, dexamethasone 8 mg, ondansetron 4 mg plus dexamethasone 8 mg or placebo (n=40 each) intravenously immediately before induction of anaesthesia. Patients were then observed for 24 hours postoperatively. The incidence of emetic episodes in the ondansetron with dexamethasone group was lower than in the placebo (p<0.001) and ondansetron (p=0.091) and dexamethasone (p=0.143) groups. A complete response (as no postoperative nausea and vomiting) was achieved in 60% of patients given ondansetron, 63% of the patients given dexamethasone, 78% of patients given ondansetron with dexamethasone and 37% of patients received placebo. The prophylactic use of ondansetron with dexamethasone is more effective in preventing postoperative nausea and vomiting.

Disseminated tuberculosis presenting as hemobilia, successfully treated by arterial embolization.

Tuberculosis, specially disseminated tuberculosis, involves the liver frequently. Focal hepatic tuberculosis with local hemorrhage has been reported. We report on a twenty-one year female with disseminated tuberculosis presenting with initially non-localisable massive upper gastrointestinal bleeding, subsequently found to have pancreatitis, right sided pleural effusion and hemobilia which was treated successfully.

Management of the Budd-Chiari syndrome by balloon cavoplasty.

BACKGROUND: Obstruction of the suprahepatic inferior vena cava (IVC) by a membrane or stricture is the commonest cause of Budd-Chiari syndrome in the eastern hemisphere. We present our experience with the outcome of balloon cavoplasty in such cases. METHODS: We followed up 40 consecutive cases of Budd-Chiari syndrome over seven years. Doppler study of hepatic venous outflow tract (in all cases), liver biopsy (30 cases) and necropsy (two cases) were performed. Balloon cavoplasty was done in selected cases. RESULTS: Of 40 patients with BCS (mean age 35.2 [SD 8.7] years; 26 men) 5, 5 and 30 had fulminant, acute and chronic presentation, respectively. Inferior vena cavography was performed in 32 cases, and showed membranous obstruction of the IVC in 12, segmental occlusion of the IVC in 11 cases, and block in both the IVC and the main hepatic veins in the rest. Successful balloon cavoplasty was done in 18 cases with obstruction of the IVC (membrane or stricture); 15 of them are well over a mean follow up of 56 (14.6) months. Three patients developed restenosis; two of them, treated with redilatation, are doing well, and one died of septicemia and hepatic failure following a surgical bypass. Pressure gradient between the IVC and right atrium decreased significantly after cavoplasty (15.4 [2.8] vs 6.6 [2.0] mmHg; p< 0.001). CONCLUSION: Balloon cavoplasty gave encouraging results in the management of Budd-Chiari syndrome due to membranous obstruction or stricture of the IVC.

Lipoprotein(a) and coronary heart disease in Indian population.

OBJECTIVE: Present study was undertaken to evaluate the role of lipoprotein(a) in coronary heart disease (CHD) patients and its relationship with other established risk factors. METHODS: Blood samples of 67 control patients (non-cardiovascular problems) and 222 CHD patients (> or = 4 weeks post myocardial infarction) were analyzed. Lipoprotein(a) was measured in serum samples by enzyme-linked immunosorbent assay utilizing rabbit polyclonal antibodies against purified human Lp(a). Step-wise linear discriminant analysis was used to find the important parameters to discriminate CHD and non-CHD subjects. RESULTS: The LDL to HDL cholesterol ratio (p < 0.01) and serum level of lipoprotein(a) (p < 0.01) were significantly higher in CHD patients. Levels of lipoprotein(a) were found to be higher in females compared to males (p < 0.01). Positive family history of CHD did not show significant difference in Lp(a) levels. Lp(a) level in CHD patients with positive family history of NIDDM and hypertension was higher than in with negative family history. CONCLUSION: Clinical significance of serum level of Lp(a) and albumin in determining the risk of CHD has been observed. Lp(a) alone could correctly discriminate a CHD individual from a control subjects by 95%. Estimating of Lp(a) together with albumin provided 99% correct discrimination between control and CHD patients. These results also suggest that Lp(a) together with malnutrition could be responsible for the increased incidence of CHD in Indians. It is also indicated that in females atherothrombogenic potential of lipoprotein(a) remains suppressed before menopause but after this stage women lose this advantage.

Assessment of normal weight in males of the age group of 40-60 years with respect to height: a new formula.

In the present work, 3000 male human subjects were studied for their health status and endeavour was made to establish the relation between the height and weight. Male individuals between the age group of 40 to 60 years were studied; their heights were between 140-180 cm. They were grouped as Group I-weight above (Height in cm-100 cm) x kg/ cm, Group II-weight between Group I and Group III subjects and Group III-weight below Group I subjects at least by 15%. It was found that Group II subjects were less sufferer from disease like, diabetics, coronary heart diseases, hypertension etc. than Group I subjects, Group III subjects were also similarly less affected by diseases but they are more prone to tiredness to accustomed work than Group II subjects. So it is concluded that the weight of Group II subjects should be considered standard.

Prevalence of diarrhoeal diseases amongst tribals of Car Nicobar Island, India.

In a cross sectional study of total 14514 tribal Nicobarese people of Car Nicobar Island, an union territory of India, situated in Bay of Bengal, morbidity, mortality and treatment practices of diarrhoeal diseases of under-five children were surveyed. The incidence of diarrhoea (0.2 episode/ch.year), Crude death rate (3.6/1000) and IMR (31/1000 live births) were exceedingly low as compared to SouthEast Asian Countries including mainland of India. ORS utilisation rate was high (41%) as compared to the National figure (12.0%). This is the first study of it's kind carried out in this island and amongst the Nicobarese tribe in India. A preliminary study carried out among the mothers of the hospitalised children indicated that protective behaviours like breast feeding, hand washing, use of chlorinated water and proper disposal of stool were regularly being practiced by the community traditionally. An in-depth study on risk factors of diarrhoea in this island can yield useful clue for describing the same.