Efecto de enzimas proteolíticas y de la abstinencia sexual en la cuantificación bioquímica de alfa glucosidasa en plasma seminal humano / Effects of proteolytic enzymes and sexual abstinence on alfa glucosidase levels in human seminal plasma

Background: a-glucosidase is found in human seminal plasma as an acid form, located in accessory glands, and as a neutral form secreted almost exclusively by the epididymis. Quantification of a-glucosidase activity is a marker of the secretory function of the epididymis and indemnity of the sperm transport pathway Aim: To obtain reference values for a-glucosidase in normal samples of seminal plasma, to evaluate its behavior in serial samples and to determine the effect of proteolytic enzymes. Material and methods: Fifty donors, with normal semen analysis according to the criteria of the World Health Organization, were evaluated. For the study with alpha-quimotrypsin, 0.1 to 10 mg/ml of the enzyme was added to the seminal plasma from a group of donors. a-glucosidase was also measured in semen obtained from nine patients at different time intervals. Results: Normal a-glucosidase values ranged from 14.52 to 25.69 µU/ml. Concentrations up to 10 mg/ml of alpha-quimotrypsin (10 times of that usually used in the liquefaction of the semen) did not alter the quantification of a-glucosidase. Serial determinations revealed oscillations in their magnitude, which stayed in each patient's characteristic range. However a subgroup presented a marked reduction of the activity of a-glucosidase as the abstinence diminished (40 percent). Conclusions: Evaluation of a-glucosidase in seminal plasma gives reliable information of the secretor state of the epididymis and especially replaces invasive methods used to evaluate the indemnity of the spermatic transport from the epididymis to the anterior urethra

Proteolytic processing of the Cyt1Ab1 toxin produced by Bacillus thuringiensis subsp. medellin

Bacillus thuringiensis produces d-endotoxins that require proteolytic processing to become active. The activation of the B. thuringiensis subsp. medellin 28 kDa (Cyt1Ab1) cytolytic toxin by trypsin, chymotrypsin and gut extract from Culex quinquefasciatus larvae was analyzed. The Cyt1Ab1 toxin of B. thuringiensis subsp. medellin was processed by all proteases tested to fragments between 23 and 25 kDa, while processing of the Cyt1Aa1 toxin produce fragments between 22.5 and 24.5 kDa. The Cyt1Ab1 toxin was preferentially processed at the alkaline pH of 12. The in vitro proteolytic processing of the Cyt1Ab1 toxin by C. quinquefasciatus larvae midgut extract showed a 25 kDa fragment; a similar result was observed when the activation was performed in the in vivo experiments. The solubilized Cyt1Ab1 toxin and the protease resistant cores generated by in vitro processing showed hemolytic activity but not mosquitocidal activity. Amino terminal sequence of the C. quinquefasciatus gut extract resistant fragment indicated that the cutting site was located between Lys31 and Asp32, with a sequence DDPNEKNNHNS; while for the trypsin-resistant fragment the cutting site was determined between Leu29 and Arg30, and for the chymotrypsin-resistant fragment between Arg30 and Lys31.

A GBP 130 derived peptide from Plasmodium falciparum binds to human erythrocytes and inhibits merozoite invasion in vitro

The malarial GBP 130 protein binds weakly to intact human erythrocytes; the binding sites seem to be located in the repeat region and this region's antibodies block the merozoite invasion. A peptide from this region (residues from 701 to 720) which binds to human erythrocytes was identified. This peptide named 2220 did not bind to sialic acid; the binding site on human erythrocyte was affected by treatment with trypsin but not by chymotrypsin. The peptide was able to inhibit Plasmodium falciparum merozoite invasion of erythrocytes. The residues F701, K703, L705, T706, E713 (FYKILTNTDPNDEVERDNAD) were found to be critical for peptide binding to erythrocytes.

Induction of proliferation in resting B-cells by a factor released by activated mouse spleen cells

Mouse spleen cells activated in a mixed lymphocyte reaction release a soluble factor, which induces a significant proliferative response in fresh mouse spleen cells. This proliferation inducing factor (PIF) was found to be heat stable (90 degrees C for 45 min) and also resistant to trypsin or chymotrypsin treatment. By using a sizing HPLC column, the molecular weight of PIF appears to be 25 kDa. Mouse spleen cells treated with anti-thy-1 + complement lost Con-A induced proliferative responses but responded well to PIF. B cell depleted spleen cells obtained by negative selection panning, did not respond to PIF. These results indicate that B cells proliferated in response to PIF. Polymixin-B, which blocks the B cell proliferative response to LPS, did not inhibit PIF induced proliferation.

Interacción de promastigotes de Leishmania donovani con celulas de exudado peritonial de ratones mediante la influencia de la quimotripsina / Interaction of Leishmania donovani promastigotes with peritoneal exudate cells of mice under the influence of chymotrypsin

Se hace un estudio de la interacción de promastigotes de Leishmania donovani con células de exudado peritoneal de ratón (c e p) mediante la influencia de la quimotripsina. La adhesión de los promastigotes a las c e p fue terminal y marginal, y en observaciones hechas a partir de los 10 minutos de enfrentamiento, esta adhesión fue nula hasta los 30 minutos en el grupo tratado, y sólo a las das horas hubo un pequeño incremento (2.4%) con respecto al control. Se observa marcada disminución en todos los parámetro medidos, tales como enlace, penetración, multiplicación intracelular, división de formas flageladas, en el grupo tratado. La quimiotripsina favorece la formación de formas intermedias flageladas, heciendose el parásito piriforme y esférico, apareciendo una forma aberrante de extremo anterior cilindrico que semeja a una forma coanoflagelada. Se sospecha que la enzima reduce efectivamente fragmentos proteicos o péptidos, los cuales pueden haber sido tan pequeños como para esconder a otros ligandos relacionados con la adhesión macrófago-parásito