Purification and properties of a decapping enzyme from rat liver cytosol.

A decapping enzyme has been purified about 2400-fold from rat liver cytosol. The decapping enzyme was shown to be fairly homogeneous by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme had an apparent molecular weight of 110,000 and consisted of two equal subunits. The enzyme hydrolyzed m7Guo5'PPP5'Ado to m7GMP and ADP. Analysis of the products produced from radioactively capped oligonucleotides and intact mRNA having 3H-cap suggests that the enzyme can hydrolyze capped mono- to pentanucleotides (m7Guo5'PPP5'N (where N = 1-5 nucleotides)) but not intact mRNA. The existence of methyl group at the N7 position of guanosine moiety of cap structure was necessary for the action of the decapping enzyme. This was confirmed by the comparison of the rates of hydrolysis of m7Guo5'PPP5'Ado by the enzyme in the presence of various nucleotides. The activity of enzyme was slightly stimulated by Na+, K+, NH4+, Ca2+ and polyamines. Mg2+ and Mn2+ were without effect on the enzyme activity.

Sexual differentiation in sensitivity to male body odor(1).

We have confirmed that more female subjects than male subjects evaluate male body odor as significantly unpleasant. Through an investigation on sexual differentiation in sensitivity to male body odor, we concluded that one of the volatile steroids, androstenone, had two effects on female olfactory sense. First, female subjects perceived androstenone itself to be more unpleasant than male subjects. Second, for only female subjects, androstenone, at a concentration of one-tenth of detection threshold, enhanced the intensity and unpleasantness of body-odor constituents such as short-chain fatty acids.

Swelling/deswelling mechanism of calcium alginate gel in aqueous solutions.

To elucidate the mechanism of dimensional changes in alginate impression in solutions, the relationship between the ion concentrations in three types of solutions (nonelectrolyte and monovalent and divalent metallic salts) and change in gel volume was examined. The gel in the monovalent metallic salt solution expanded and a decrease in monovalent cation and an increase in Ca2+ were observed. This suggests that the crosslinking density of the gel reduced due to dissociation of Ca2+ from the calcium alginate gel. In divalent metallic salt solutions, the shrinkage occurred according to crosslinking of unreacted residue. In the nonelectrolytic solution, gel, neither ethylene glycol nor acetonitrile showed any volume changes, while that in glutaraldehyde contracted. It is speculated that the former two solutions were affected by the fact that the gel has no semipermeability, and that the latter result was due to chemical interaction between the gel and solution.

The effect of evolution on host-parasitoid systems.

It is well known that a simple first-order difference equation can exhibit complex population dynamics, such as sustained oscillations and chaos. An interesting problem is whether such oscillatory dynamics are expected to occur in real populations. This paper assumes that the resident system is composed of 1-host and 1-parasitoid and that only the host is allowed to evolve, but not the parasitoid. Based on the invasibility of a host to host-parasitoid systems, we investigate the dynamics of the host-parasitoid system favored by natural selection. We consider two cases. In the first case, the host's evolution involving both the intrinsic growth rate and the sensitivity to density is considered. In the second case, the host's evolution involving both the intrinsic growth rate and the vulnerability to the parasitoid is considered. In both cases, we see that the dynamics with a stable equilibrium will not be favored by natural selection without the trade-off between the host's traits which are allowed to evolve. The host-parasitoid system with a stable equilibrium will be eventually invaded by a host type that develops an unstable equilibrium with the parasitoid. If there is a trade-off between the host's traits which are allowed to evolve, a host-parasitoid system with a stable equilibrium can be favored by natural selection.

Artificially damaged hairs: preparation and application for the study of preventive ingredients.

We have developed a method for preparing artificially damaged hairs, similar to those generally observed in permed hair. Moreover, we have established two models of hair for testing preventive ingredients. A model for scale lift was prepared using alkaline protease digestion following lyophilization. A model for splitting, was prepared by successive extraction of the cortical protein, re-oxidation, and lyophilization. The practical application of these models was confirmed through the evaluation of the preventive effect of a polymer or peptides.

Metabolism of paeonol in rats.

As a part of our studies on the metabolism of active components from traditional Chinese medicines, paeonol was orally administered to rats. The urinary metabolites were analyzed by 3D HPLC, and their structures were determined to be 2, 4-dihydroxyacetophenone-5-O-sulfate (P1), resacetophenone-2-O-sulfate (P2), 2-hydroxy-4-methoxyacetophenone-5-O-sulfate (P3), paeonol-2-O-sulfate(P4), resacetophenone (P5), and unchanged paeonol, on the basis of their chemical and spectral data. Among these metabolites, P2-P4 and paeonol were detected in the plasma after the oral administration of paeonol. Furthermore, the bile of rats given paeonol orally was found to contain P3, suggesting the enterohepatic circulation of paeonol.

An h.p.l.c. method for the quantitative determination of asulam, acetylasulam and sulphanilamide in peaches.

An analytical method has been developed for the determination of the herbicide, asulam [methyl(4-aminobenzenesulphonyl)carbamate] and two metabolites, sulphanilamide and acetylasulam individually, in peaches. Asulam and acetylasulam were partitioned from an aqueous phase with ethyl ether, leaving behind more polar components which interfered with the analysis of these compounds. The sulphanilamide was next partitioned from the aqueous phase into ethyl acetate. Each fraction was then chromatographed on its own neutral alumina column, activity grade 1, and analysed by high performance liquid chromatography (h.p.l.c.) using a C-18 reverse phase column. From peach samples fortified at the 0.10 p.p.m. level, recoveries averaged 72% (asulam), 100% (acetylasulam) and 90% (sulphanilamide). No residues of these compounds were found in two studies of peaches harvested 0, 5, 10, 15 and 20 days following asulam treatment at 0-7.50 kg/ha.

Coagulation factor XI is a contaminant in intravenous immunoglobulin preparations.

A small number of thromboembolic events, including deep venous thrombosis and myocardial infarction, have been reported in patients receiving IVIG. These events have primarily occurred in patients receiving high-dose IVIG and have been attributed to an increase in blood viscosity. To test the hypothesis that a procoagulant might be present in IgG preparations, twenty-nine samples of intravenous immunoglobulin (IVIG) from eight different manufacturers were assayed for procoagulant activity. Twenty-six of these samples shortened the clotting time of factor XI-deficient plasma. Of these, fourteen samples had factor XI activities greater than 0.001 U/ml of normal pooled plasma. The remaining samples possessed less than 0. 001 U/ml of normal plasma activity. The procoagulant activity in these samples could be inhibited by an anti-factor XI polyclonal antibody, suggesting that the procoagulant activity was factor XI. The procoagulant activity increased in two samples after storage at 4 degrees C for 4 weeks, likely as a result of factor XIa autoactivation. Additionally, activity in some IVIG samples was able to directly activate factor IX, indicating that activated factor XI was present in these samples. Finally, the degree of factor XI(a) contamination in the samples was correlated with the manufacturer, suggesting that variations in the manufacturing process or source plasma affect the level of factor XI in the IVIG product. Because addition of small amounts of factor XIa to plasma can lead to production of significant amounts of thrombin, we suggest that factor XIa present in some IVIG preparations could contribute to the in vivo risk of thrombosis after IVIG therapy.

Deencryption of cellular tissue factor is independent of its cytoplasmic domain.

Tissue factor (TF) is a transmembrane molecule that, when exposed to plasma, is the key initiator of coagulation. Cellular TF activity is normally "encrypted", but treating cells with calcium ionophore (i.e. , ionomycin or A23187) increases ("deencrypts") TF activity without increasing TF mRNA or antigen expression. Deencryption results from both plasma membrane phosphatidylserine (PS)-dependent and -independent mechanisms; however, the nature of the PS-independent component is unclear. Since deencryption has been suggested to result from release of TF dimers on the cell surface, and since TF's cytoplasmic domain binds to actin-binding protein 280 and interacts with the cytoskeleton, we hypothesized that interactions with the cytoskeleton, through the cytoplasmic domain, play a role in mediating encryption/deencryption. We examined TF deencryption and the role of the cytoplasmic domain in the PS-independent component using baby hamster kidney (BHK) cells expressing full length TF (BHK-TF) or TF lacking its cytoplasmic domain (BHK-descyt) (Sorensen et al. (1999) J. Biol. Chem. 274, 21349). Both BHK-TF and BHK-descyt cells exhibited a dose-dependent, 1.5- to 10-fold increase in TF activity upon treatment with calcium ionophore, and this increase in activity was only partially blocked by annexin V. These results indicate that deencryption is not restricted to cells which naturally express TF and that the PS-independent component of deencryption is intact on cells transfected with either full length or truncated TF. Our results clearly indicate that deencryption is not dependent on an intact cytoplasmic domain in transfected BHK cells.

[Changes in the electrical activity of the cerebral cortex under the combined influence of Cl. perfringens type A toxin and products of Cl. butyricum activity]. / Izmeneniia élektricheskoi aktivnosti kory golovnogo mozga pri kombinirovannom deistvii toksina Cl. perfringens tipa A i produktov zhiznedeiatel'nosti Cl. butyricum

A study of the changes of the electrocorticogram (ECoG) and the electrocardiogram (ECG) in combined action of Cl. perfringens, type A, and of the Cl. butyricum broth culture filtrate showed that desynchronization of the cortical electrical activity and its subsequent depression occurred at earlier periods than in the case of isolated administration of Cl. perfringens toxin. The general character of the changes in the cortical rhythmic activity remained the same as in intoxication caused by Cl. butyricum toxin alone. The ECoG and ECG changes occurred at shorter intervals. Cl. butyricum filtrates induced no ECoG and ECG changes. It is supposed that the effect of the products of the Cl. butyricum vital activity consisted in increase in the tissue barrier permeability and, in this connection, in a greater penetration of Cl. perfringens toxin into the tissues, including the central nervous system.