Purification and tissue/species dependence of the specificity of buffalo kidney cathepsin B.

A simple purification scheme was developed for isolation and purification of cathepsin B from buffalo kidney. The use of CM-Sephadex and chromatofocusing helped in better and simultaneous separation of cathepsin B, H and L. As judged by PAGE and SDS-PAGE studies, the enzyme was found to be pure on the basis of charge and had a molecular mass of 25.5 kDa. The amino acid composition, number of free sulfhydryl groups and other major physico-chemical properties of the purified enzyme were similar to the properties reported for cathepsin B from other sources/tissues. However, the NH2-terminal amino acid residue of the enzyme was found to be Ala as against Leu reported from other tissues/species. The total carbohydrate content was also found to be significantly lower (3.6%) as compared to 7.0-7.6% reported for the enzyme from other sources. Thiol reducing compounds activated the enzyme whereas thiol blocking compounds inhibited it. The buffalo kidney enzyme hydrolyzed Z-Phe-Arg-MCA (Vmax/K(m) = 17.1) as the most efficient substrate followed by Z-Arg-Arg-MCA, BANA and BAPNA. Among the protein substrates, goat hemoglobin (Vmax/K(m) = 874) was found to be the most preferred. Rabbit muscle aldolase, usually considered to be a good substrate for cathepsin B, proved to be a poor substrate for this enzyme; only 25-30% inactivation of aldolase was observed. Antibodies raised against the enzyme recognised only cathepsin B and did not have any cross reactivity with cathepsin H or L from the same or different sources. These differences in the properties of the buffalo kidney enzyme vis-a-vis the same enzyme from other tissue/species have been attributed to specialized function of cathepsin B in diversified tissues.

Hyperprolactinemia and reproductive disorders--a profile from north east.

Serum prolactin was estimated in 90 women of various menstrual disorders and infertility. Hyperprolactinemia (PRL 25 ng/ml) was detected in twenty four cases. Out of these, nine had prolactinoma, four had hypothyroidism, three had drug induced HPRL and in eight, no cause could be ascertained. Five patients with prolactinoma became pregnant following bromocryptine treatment and delivered normal babies.

Mass closure versus layer closure of abdominal wound: a prospective clinical study.

The results of a controlled clinical trial of two currently recommended methods of abdominal closure have been reported here. OUt of 160 patients, 80 cases were randomised to have their abdominal wall closed in single layer mass closure with monofilament nylon and 80 cases in layer closure with chromic catgut. Eighteen patients (22.5%) in the former and 38 (47.5%) in the latter had wound infection (p < 0.001). Sinus formation occurred in 2 patients (2.5%) in the mass and 20 (25%) in the layered group (p < 0.001). Three cases (3.75%) of burst abdomen occurred in layer closure and none in mass closure. Wound infection was the most important denominator next to suture material influencing wound healing. There was a significant association between the rate of infection and sinus formation. Minimal complication and good patient compliance seem to justify the use of mass closure in place of layer closure in all types of abdominal operations.