Changes in the electrophoretic pattern of the venom of the bushmaster (Lachesis muta stenophrys) stored under various conditions

Liquid and lyophilized samples of Lachesis muta venom were stored at different temperatures and for different periods of time, and analyzed by polyacrylamide gel electrophoresis (PAGE) and immunoelectrophoresis. Only slight variations were evident when three pools of freeze-dried venom, that had been kept at -30 degrees C for several months were compared with fresh venom. These results suggest that L. muta venom is not altered drastically when stored under these conditions.     

Study of stereoisomers of N6-phenylisopropyladenosine on the secretion of pancreatic glucagon

This work was designed to characterize the adenosine receptor (A1 or A2) involved in glucagon secretion. The most potent adenosine analogues on A1 receptors are the N6 substituted compounds, among them N6-phenylisopropyladenosine (PIA); furthermore L-PIA is 50 to 100 times more potent than D-PIA on the A1 receptor, whereas it is 3 to 5 times more potent on the A2 receptor; thus the A1 receptor shows a much higher stereoselectivity. The effects of L-PIA and D-PIA were studied on glucagon secretion from the isolated perfused rat pancreas. 1) L-PIA at 1.65 microM induced a transient glucagon secretion which was not greater than that induced by the same concentration of adenosine. 2) D-PIA at a 3 fold higher concentration (4.95 microM) elicited a secretion of glucagon comparable to that induced by L-PIA 1.65 microM; thus the involved receptor does not present a high stereoselectivity for L-PIA. These results support the fact that the receptor involved in glucagon secretion is not of the A1 type.     

Structural aspects of intranuclear matrix disintegration upon RNase digestion of HeLa cell nuclei

The organization of the intranuclear elements observed in histone-depleted (2 M NaCl-extracted) HeLa cell nuclei was investigated by means of electron microscopy and two-dimensional gel electrophoresis. This work was mainly aimed at verifying whether or not an intranuclear skeleton or matrix existed, which could explain the stable attachment of RNA to the residual nuclear structure after high-salt extraction, and its three-dimensional organization. We compared the ultrastructure and the polypeptide composition of RNA-containing and RNA-depleted (RNase-treated) nuclear residues, and we visualized intermediate stages of RNase action on the intranuclear material. We showed that this material was made of two types (fibrillar and granular) of salt-resistant RNP components equally sensitive to RNase when the enzyme was used prior to high-salt extraction. At least in our material and under our experimental conditions, no intranuclear matrix could be distinguished from the residual RNP material. Our results further suggest that formation of such a matrix is a path-dependent phenomenon.     

All out anaerobic capacity tests on cycle ergometers. A comparative study on men and women

We have studied the effects of the braking force on the results of an anaerobic capacity test derived from the Wingate test (an all out 45 s exercise on a Monark 864 cycle ergometer against a given force at the fastest velocity from the beginning to the end of the test). Seven men and seven women participated in the study and performed a total of 63 all-out tests against different braking forces. The same subjects performed a force-velocity test on the same cycle ergometer. Since the relationship between force and velocity is approximately linear for peak velocities between 100 and 200 rev X min-1 (Pérès et al. 1981a, b; Nadeau et al. 1983; Vandewalle et al. 1983) we characterized each subject by three parameters: P0 (the intercept of the force-velocity regression line with the force axis), V0 (the intercept of the regression line with the velocity axis) and Wmax (maximal power). The relationship between force and mean power was parabolic for the all-out anaerobic capacity test. In the present study the optimal force (the force giving the maximal value of mean power during an all out test) was higher for the men (approximately 1 N X kg BW-1) than the force proposed by others (0.853 N X kg BW-1 for Dotan and Bar-Or 1983). However, because of the parabolic relationship between force and mean power, the mean power which corresponds to the optimal force was approximately the same in both studies.(ABSTRACT TRUNCATED AT 250 WORDS)     

Enzymatic hydrolysis of asymmetric heterocyclic amino acid derivatives

Summary 2-methyl-oxazolinones and thio-thiazolidones representative of aromatic and aliphatic amino acids were hydrolysed by -chymotrypsin and subtilisin. The parametric ratio k_cat/K_m, correlated with the enantiomeric enrichment of the reaction product, indicates that thio-thiazolidones are converted to free amino acids by enzyme with the higher degree of stereospecificity.     

Evidence for the participation of disulfide and sulfhydril groups in the specific binding of [3H]prazosin in cerebral cortex

The tritiated ₁ antagonist prazosin [³H]PRZ binds specifically and with high affinity to postsynaptic adrenoceptors in membrane preparations from cerebral cortex. Since adrenoceptors are of protein nature, it was of interest of investigate the possible role of disulfide (—SS—) and sulfhydril (—SH) groups in the binding of [³H]PRZ. Pretreatment of the membranes with the disulfide and sulfhydryl reactivesdl0Dithiothreitol,l-Dithiothreitol, Dithioerythritol or 5,5-Dithiobis-(2-nitrobenzoic acid) (DTNB), alone or in combination with the alkylating agent N-Methylmaleimide (NMM), decreased specific [³HPRZ binding, with minor changes in the non-specific counts. Saturation experiments revealed that all these reagents reduced the affinity of the binding site for [³H]PRZ, as judged by theK _d 25°C, but only the alkylating agent NMM and the oxydizing reagent DTNB produced in addition to the increase inK _d, a decrease of the maximum binding capacity (B _max). The present results provide evidence for a participation of—SS—and/or—SH groups in the recognition site of the ₁-adrenoceptor of cerebral cortex.     

Lysosomal hydrolase activity in chorionic villi and embryonic cells in culture

Summary Fourteen lysosomal enzymes were compared in 20 cultured cell lines from chorionic biopsy and corresponding embryonic tissue after voluntary abortions. Enzymatic expression appears to be similar in cultured cells from both sources with some slightly higher levels for chorionic villi. We stress the importance of culturing chorionic villi especially in the case of enzymes (-L-iduronidase) or diseases (I cell disease) whose expression is unusual in fresh trophoblast tissue.     

Repair of damage in double-stranded phi X174 (RF) DNA due to radiation-induced water radicals

Experiments in which the yields of radiation-induced OH and H radicals were varied, showed that both types of water radicals inactivate phi X174 RF DNA to about the same extent as measured by transfection of the (irradiated) DNA to E. coli wild-type spheroplasts. On the other hand, using spheroplasts prepared from E. coli strains, deficient in one of the proteins involved in excision DNA repair (uvrA- or uvrC-) or in post-replication repair (recA-), clear differences between damage originating from OH or H radical attack were found. Part of the radiation damage due to H radicals appeared to be repairable by an uvrA-gene-dependent repair mechanism, whereas this repair pathway does not play an important role in the case of OH radical damage. The reverse applies to uvrC-gene-dependent repair, which only affects OH radical damage (obtained under anoxic conditions), but has no influence on damage due to H radicals. Irradiation of double-stranded phi X174 (RF) DNA in the presence of oxygen however, yields damage--due to OH radicals only--which appeared not to be sensitive to either uvrC- or uvrA-gene-dependent repair. Furthermore, post-replication repair (recA) has only very little effect on the amount of inactivation by H or OH radicals, when irradiation is carried out under anoxic conditions. We did not find significant inactivation due to hydrated electrons, whether the biological activity was determined by use of wild-type spheroplasts or of strains deficient in excision or post-replication repair proteins.     

Binding of 5-methoxypsoralen to human serum low density lipoproteins

5-methoxypsoralen (5-MOP) binds to human serum low density lipoproteins (LDL) according to a two-step process. Scatchard analysis of the first step yields K = 1.4 × 10⁵ M^?1 and 4 binding sites. It involves the LDL apoprotein. The second step corresponds to a solubilization, in the lipidic core, of ? 45 molecules of 5MOP per LDL molecule. It is accompanied by a large blue shift of the 5MOP fluorescence. The ability of LDL to bind 5MOP and to carry it into various cells may explain some biological effects sometimes encountered during PUVA therapy.     

Natural killer activity of kurloff cells: A direct demonstration on purified kurloff cell suspensions

In order to study their natural killer effect, guinea pig splenic Kurloff cells were fractionated by Percoll discontinuous density gradient centrifugation. Kurloff cells were collected and tested for cytotoxicity in a 24-hr chromium-release test. Comparison of different splenic cellular samples (of males or estrogenized females) with increasing percentage of Kurloff cells, revealed a highly significant positive correlation (r = 0.93, α < 0.01) with the cellular cytotoxicity developed against the K 562 target cells.