A comparative study on cell disruption methods for release of aspartase from E. coli K-12.

Applicability of different mechanical cell disruption techniques namely sonication, bead milling and French press for the release of aspartase from E. coli K-12 was compared. Various operating parameters of each technique were optimized to obtain maximum aspartase release. The efficiency of aspartase release and cell disruption by all the methods was also compared under optimal conditions. The maximum release of aspartase (98.22%) and maximum cell breakage (84.25%) was observed using French press, while 92% of aspartase release was obtained by both sonication and bead milling. The order of cell disruption constant (k) for aspartase release by these methods was French press > bead milling > sonication. Disruption of cells using French press also demonstrated maximum protein release (14.12 mg/mL). The crude enzyme preparations can be further used for purification and its applications.

Characterization and antimicrobial activity of lectins from Penicillium sp.

Ten Penicillium sp. were screened for lectin activity for occurrence of lectins. Mycelial extracts from submerged cultures of P. corylophilum, P. expansum and P. purpurogenum showed agglutination against human (A, B, AB and O), goat, sheep, pig and rabbit erythrocytes. Neuraminidase treatment to human blood type O erythrocytes substantially increased their agglutinability by all the lectins as compared to untreated erythrocytes. Modification of erythrocyte surfaces by protease increased the lectin titre only of P. corylophilum with no effect on other two lectins. P. corylophilum and P. expansum displayed relatively lower titres in mycelial extracts prepared from agar plate cultures as compared to broth cultures. A panel of sugars was tested for inhibition of lectin activity. All the lectins were found to be specific for asialofetuin, bovine submaxillary mucin, porcine stomach mucin, chondroitin-6-sulphate, D-sucrose and D-glucose. P. corylophilum lectin was expressed (Titre 8) by 5 day old cultures, reaching its maximum level (Titre 32) upon 8 days of cultivation, thereafter declin in lectin activity was observed. P. purpurogenum lectin was expressed by 7-10 days old cultures, while in P. expansum maximum lectin activity was elaborated by 5-8 days old cultures. Lectin extracts from all the three species were found to possess antimicrobial activities. Lectin extracts from the three Penicillium species displayed antifungal activity and antibacterial activity against Gram-negative and Gram-positive bacterial strains.

Antibiotic resistance and pathogenicity factors in Staphylococcus aureus isolated from mastitic Sahiwal cattle.

Methicillin-resistant Staphylococcus aureus (MRSA) poses a serious problem in dairy animals suffering from mastitis. In the present study, the distribution of mastitic MRSA and antibiotic resistance was studied in 107 strains of S. aureus isolated from milk samples from 195 infected udders. The characterizations pathogenic factors (adhesin and toxin genes) and antibiotic susceptibility of isolates were carried out using gene amplification and disc diffusion assays, respectively. A high prevalence of MRSA was observed in the tested isolates (13.1%). The isolates were also highly resistant to antibiotics, i.e. 36.4% were resistant to streptomycin, 33.6% to oxytetracycline, 29.9% to gentamicin and 26.2% each to chloramphenicol, pristinomycin and ciprofloxacin. A significant variation in the expression of pathogenic factors (Ig, coa and clf) was observed in these isolates. The overall distribution of adhesin genes ebp, fib, bbp, fnbB, cap5, cap8, map and cna in the isolates was found to be 69.1, 67.2, 6.5, 20.5, 60.7, 26.1, 81.3 and 8.4%, respectively. The presence of fib, fnbB, bbp and map genes was considerably greater in MRSA than in methicillin-susceptible S. aureus (MSSA) isolates. The proportions of toxin genes, namely, hlb, seb, sec, sed, seg and sei, in the isolates were found to be 94.3, 0.9, 8.4, 0.9, 10.2 and 49.5%, respectively. The proportions of agr genes I, II, III and IV were found to be 39.2, 27.1, 21.5 and 12.1%, respectively. A few isolates showed similar antibiotic-resistance patterns, which could be due to identical strains or the dissemination of the same strains among animals. These findings can be utilized in mastitis treatment programmes and antimicrobials strategies in organized herds.

Inflammatory pseudotumour of the lung : report of a case and review of literature.

A case of unusual benign tumour of the lung is described. The patient presented with a history of prolonged cough with expectoration, and fever with generalised weakness of shorter duration. Radiological examination of the chest revealed a large non-homogeneous opacity with calcification in the left lower zone. Fine needle aspiration cytology (FNAC) was inconclusive. Therefore, surgical exploration and a left lower lobectomy was performed. The diagnosis of inflammatory pseudotumour was made on histopathological examination.

In vitro regeneration of Hypericum patulum Thunb.--a medicinal plant.

A protocol was developed for high frequency plant regeneration in H. patulum by shoot-tip culture. H. patulum plants were collected from a wild source growing at high altitude in the eastern Himalayas. Multiple buds were initiated from shoot-tips cultured on Murashige and Skoog's medium supplemented with BAP, kinetin. Addition of thiamin HCI, Ca-pantothenate and biotin enhanced multiple shoot formation. Upon transfer to phytohormone free liquid medium following a brief exposure to auxin, root formation occurred from the micro shoots . Rooted plants were hardened and transferred to soil. Regeneration potentiality was found to be constant throughout the year in long term cultures.

A comparison of esmolol and diltiazem for heart rate control during coronary revascularisation on beating heart.

This prospective study compared control of heart rate and haemodynamics during coronary artery revascularization without cardiopulmonary bypass using either esmolol or diltiazem. Sixty adult patients with one or two vessel coronary artery disease, were randomly divided into 2 groups. Group A (n=30) received a 50 microg/kg/ loading dose of esmolol followed by a 100 microg/kg/hr infusion, for control of heart rate during surgical anastomosis of the coronary vessel. Group-B (n=30) received 0.15 mg/kg of diltiazem as a loading dose followed by a 5 mg/hr infusion for heart rate control, during the anastomosis. It was seen that heart rate control was better in group A, 51.4 +/- 1.3 beats/min, (p <0.01) than in group B, 69.6 +/- 3.0 beats/min (p <0.05), as compared to baseline values of 80.6 +/- 12.1 beats/min in group A and 82.4 +/- 10.6 beats/min in group B respectively. Systemic vascular resistance and pulmonary artery wedge pressure were unchanged in group A but mean pulmonary artery pressure and pulmonary vascular resistance were significantly raised. Group B patients had decreased systemic vascular resistance, mean pulmonary artery pressure and pulmonary artery wedge pressure, and reduced right ventricular stroke work index at the time of distal coronary anastomosis. We concluded that although esmolol provided dramatically slower heart rates, during surgery, the resulting elevations in mean pulmonary artery pressure and pulmonary vascular resistance would require caution if used in patients with underlying right ventricular dysfunction from ischaemia or infarction. Diltiazem by virtue of its effects on systemic vascular resistance, cardiac output, and lowering of mean arterial pressure may be a better choice in hypertensive patients.

A comparison of esmolol & diltiazem for heart rate control during coronary revascularisation on beating heart.

This prospective study was done to compare the control of heart rate and haemodynamics during coronary artery revascularisation without cardiopulmonary bypass using either esmolol or diltiazem. Sixty adult patients with one or two vessel coronary artery disease, were randomly divided into 2 equal groups. Group A received a 500 micrograms/kg loading dose of esmolol followed by a 100 micrograms/kg/h infusion, for control of heart rate during surgical anastomosis of the coronary vessel. While Group B received 0.15 mg/kg diltiazem as a loading dose followed by a 5 mg/h infusion for heart rate control, during the anastomosis. It was seen that heart rate control was better in Group A, 51.4 (+/- 1.3) beats/min, than in Group B, 69.6 (+/- 3.9) beats/min but the decrease in heart rate was significant in both the groups at peak effect compared to respective predrug values. Group A patients had unchanged systemic resistance and pulmonary artery wedge pressure but mean pulmonary artery pressure and pulmonary vascular resistance were significantly raised. Group B patients had decreased systemic resistance, mean pulmonary artery pressure and pulmonary artery wedge pressure, and reduced right ventricular stroke work index. We concluded that although esmolol provided dramatically slower heart rates, during surgery, the resulting elevations in mean pulmonary artery pressure and pulmonary vascular resistance would require caution if used in patients with underlying right ventricular dysfunction from ischaemia or infarction. Diltiazem by virtue of its effects on systemic vascular resistance, cardiac output, and lowering of mean arterial pressure may be a better choice in hypertensive patients.

Unusual intrapulmonary foreign body: a pencil.

A case of unusual intrapulmonary foreign body in the form of a pencil is described. The penetration occurred following an insignificant fall. The diagnosis was made on CT scan and subsequently surgical removal was undertaken successfully.

Tropical pulmonary eosinophilia presenting as eosinophilic pleural effusion.

A case of tropical pulmonary eosinophilia presenting as an eosinophilic pleural effusion is reported. The condition resolved by treatment with diethylcarbamazine. The patient has been well during the two year follow-up period.

Cutaneous anthrax--a report of ten cases.

The clinical features of ten cases of cutaneous anthrax are reported. Six males and four females were affected. Their ages ranged from one and a half year to sixty years. The average incubation period was 8 days. Fever and headache were common systemic manifestations. Pruritus and ulcer with formation of black eschar were typical features. The patients were treated with penicillins and chloramphenicol. There was no mortality. Possible human-to-human spread in a patient is discussed.

Outbreak of human anthrax in Ramabhadrapuram village of Chittoor district in Andhra Pradesh.

Thirty human anthrax cases were reported from Ramabhadrapuram village of Chittoor district in Andhra Pradesh during November-December, 1989. These cases occurred following an epizootic of anthrax among cattle and sheep of the village and ingestion of contaminated meat by the villagers. The overall attack rate was 24.39 per cent with a case fatality of 16.67 per cent. All age groups and both sexes were affected. Ten cases were of cutaneous form with typical black eschar formation which were confirmed bacteriologically. Fever and headache were common systemic manifestations. They responded well to penicillins and there was no mortality. The possibility of human to human spread is suggested. The twenty cases of internal anthrax comprised intestinal, septicemic, peritonitis, meningeal and pulmonary forms. Sub-clinical forms also occurred. Fever, abdominal pain, ascites, anorexia and vomiting were notable features. Diagnosis was made clinically and also on epidemiological basis. All deaths during this outbreak occurred in women with internal anthrax, the case fatality rate for the latter being 25 per cent. Prophylactic administration of penicillin was done for individuals at risk.

Retinal hemorrhages in newborn--fetal causative factors.

Two hundred and eight alive newborns examined within 2 hours of birth showed 37.50% incidence of retinal hemorrhage (RH). Out of these, 38.59% were full term and 29.17% were preterm. Head circumference had a significant correlation (p less than 0.05) with retinal hemorrhage, while sex of the newborn, birth weight, gestational age and fetal distress showed no significant correlation. Birth injuries especially ocular and periocular injuries showed a significantly higher incidence of retinal hemorrhage (83.33%).