Hexon-only binding of VP26 reflects differences between the hexon and penton conformations of VP5, the major capsid protein of herpes simplex virus.

VP26 is a 12-kDa capsid protein of herpes simplex virus 1. Although VP26 is dispensable for assembly, the native capsid (a T=16 icosahedron) contains 900 copies: six on each of the 150 hexons of VP5 (149 kDa) but none on the 12 VP5 pentons at its vertices. We have investigated this interaction by expressing VP26 in Escherichia coli and studying the properties of the purified protein in solution and its binding to capsids. Circular dichroism spectroscopy reveals that the conformation of purified VP26 consists mainly of beta-sheets (approximately 80%), with a small alpha-helical component (approximately 15%). Its state of association was determined by analytical ultracentrifugation to be a reversible monomer-dimer equilibrium, with a dissociation constant of approximately 2 x 10(-5) M. Bacterially expressed VP26 binds to capsids in the normal amount, as determined by quantitative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cryoelectron microscopy shows that the protein occupies its usual sites on hexons but does not bind to pentons, even when available in 100-fold molar excess. Quasi-equivalence requires that penton VP5 must differ in conformation from hexon VP5: our data show that in mature capsids, this difference is sufficiently pronounced to abrogate its ability to bind VP26.     

Can winter swimming cause people to prefer lower room temperatures?

Preferred room temperatures were determined for 16 winter swimmers who had been exposed to cold shock daily by bathing in the sea (0–6°C) during at least two winter seasons. Each subject participated in one comfort experiment (2 1/2 h) in an environmental test chamber, where the preferred ambient temperature was determined by adjusting it according to his wishes. The subjects were sedentary and clothed in a standard uniform. Skin temperatures, rectal temperature and evaporative weight loss were measured. The results were compared with similar data for two other groups: (a) a control group of 64 normal subjects and (b) 16 persons who had been working 8 h a day at 8°C during at least 6 months in the meat packing industry. The results showed no significant differences between the ambient temperatures preferred by the three groups of subjects. This indicated that man cannot be adapted to prefer lower ambient temperatures by exposing him daily to cold.     

Ultrastructural observations on marine choanoflagellates (Choano-flagellida, Acanthoecidae) from the coast of Thailand: species of Apheloecion gen.nov

Three species of loricate choanoflagellates (Choanoflagdlida,Acanthoecidae), collected from the Andaman Sea near Phuket Island(SW Thailand), have been described and referred to Apheloeciongen.nov. (holotype: A. quadrispinum sp.nov.). All species possessa single-chambered lorica composed of one transverse costs overlaidby a limited number of longitudinal costae which converge posteriorly.Anteriorly the longitudinal costae protrude above the transversecosta as sharp pointed spines. In A. quadrispinum sp.nov. andA. pentacanthum sp.nov. the lorica is terminated by a shortposterior spine, whereas in A. articulatum sp.nov. the pedicelis much more prominent, consisting of several costal stripswhich are joined end-to-end. The species of Apheloecion appearto be most closely related to species of Calliacantha and Monocosta.None of the species described are so far known from localitiesoutside the Andaman Sea.     

Quantification of Y chromosome bearing spermatozoa of cattle using in situ hybridization.

An easy assay for quantification of Y chromosome-bearing sperm (Y-sperm) is needed, especially to monitor sperm separation techniques. In the present study a tritiated bovine male-specific DNA fragment was tested for identification of Y-sperm by in situ hybridization. A protocol for in situ hybridization to bovine sperm was developed and used to study the proportion of Y-sperm of 12 bulls. The usefulness of the method in optimization of sperm separation procedures is illustrated through analysis of fractions of sperm separated by Percoll density gradient centrifugation.     

A nuclear magnetic resonance spectroscopic investigation of Kdo-containing oligosaccharides related to the genus-specific epitope of Chlamydia lipopolysaccharides.

The 1H- and 13C-NMR parameters, chemical shifts and coupling constants, for the pentasaccharide of the genus-specific epitope of Chlamydia lipopolysaccharide and related di-, tri-, and tetra-saccharides have been measured and assigned completely using 1D and 2D techniques, and their structures have been confirmed. NOE experiments indicated the preferred conformation of the pentasaccharide and the component oligosaccharides. The 3JH,H demonstrate a change in conformation by rotation of the C-6-C-7 bond of the side chain of the (2----8)-linked Kdo (unit b) in alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcN-(1--- -6)- GlcNol, alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcNAc-(1- ---O)- allyl, and alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----O)-allyl relative to that preferred in alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcNAc-(1----O)-allyl, alpha-Kdo-(2----8)-alpha-Kdo-(2----O)-allyl, alpha-Kdo-(2----4)-alpha-Kdo-(2----O)-allyl, and alpha-Kdo-(2----6)-beta-GlcNAc-(1----O)-allyl, irrespective of the size of the aglycon, e.g., allyl or beta-D-GlcN residues. The conformational results have been substantiated by computer calculations using the HSEA approach.     

Species differences in the negative inotropic effect of acetylcholine and soman in rat, guinea pig, and rabbit hearts.

1. Acetylcholine reduced atrial contractions by 82.5% in guinea pig, 50.8% in rat, and 41.5% in rabbit. 2. The EC50 values for the negative inotropic effect of acetylcholine were 3.3 x 10(-7) M in rat and guinea pig atria and 4.1 x 10(-6) M in rabbit atria. 3. There was no correlation between the species differences in the negative inotropic effect of acetylcholine in atria and the density or affinity of acetylcholinesterase or muscarinic receptors. 4. Inhibition of atrial acetylcholinesterase with soman reduced the EC50 of acetylcholine three-fold in all species, but did not change the maximal inotropic effect of acetylcholine. 5. Species differences in the negative inotropic effect of acetylcholine may be caused by differences in the coupling between myocardial muscarinic receptors and the ion channels that mediate negative inotropy.     

Isolation and characterization of alpha-amylase messenger RNA from bank vole parotid glands. Evidence for two separate messenger RNAs coding for amylase and an amylase-related protein

Bank vole saliva contains two glycogen-precipitable proteins, both of which show affinity for the alpha-amylase inhibitor cycloheptaamylose. One of these proteins, amylase, has a molecular weight of 55,000, judged from dodecylsulphate/acrylamide gel electrophoresis. The other has an apparent molecular weight of 59,000 and has no amylase activity. We report here that tryptic peptide maps as well as amino-acid composition analyses indicate extensive homology between the two proteins. We have also isolated total poly(A)-containing mRNA from amylase-rich bank vole parotid glands. These mRNAs were translated in the presence of [35S]methionine in an mRNA-dependent cell-free translation system from rabbit reticulocyte lysate. The radioactive translation products were examined by dodecylsulphate/polyacrylamide gel electrophoresis. Two major translation products with apparent molecular weights of approximately 56,500 and 60,500, respectively, were further characterized by tryptic peptide analyses. Our data indicate that the 56,500-Mr product is the biosynthetic precursor of amylase, whereas the 60,500-Mr translation product is a precursor of the 59,000-Mr amylase-like protein. Both precursors appear to contain extra peptide material, presumably as amino-terminal 'pre' or 'signal' peptides, in analogy with that found for other precursors of secretory proteins. Thus, amylase and the 59,000-Mr protein, although very similar, are translated from two separate mRNAs. These two messengers sediment in a sucrose gradient at about 17-S, corresponding to lengths of about 1,800 nucleotides.     

Selective inhibition of human acetylcholinesterase by galanthamine in vitro and in vivo

We investigated the inhibition of human cholinesterases by galanthamine, an alkaloid of the common snowdrop (galanthus nivalis). In vitro, the compound showed potent enzyme inhibition and 50-fold selectivity for acetylcholinesterase (EC 3.1.1.7) as opposed to butyrylcholinesterase (EC 3.1.1.8). There was no difference between enzyme inhibition by galanthamine in whole blood and separated fractions of plasma and erythrocytes. We conclude that galanthamine does not accumulate in large amounts in red blood cells. In vivo, administration of galanthamine in a healthy volunteer and in a patient who underwent long-term treatment confirmed the selectivity of galanthamine for acetylcholinesterase.