Localization and synthesis of an antigenic determinant of herpes simplex virus glycoprotein D that stimulates the production of neutralizing antibody.

An antigenic determinant capable of inducing type-common herpes simplex virus (HSV)-neutralizing antibodies has been located on glycoprotein D (gD) of HSV type 1 (HSV-1). A peptide of 16 amino acids corresponding to residues 8 to 23 of the mature glycoprotein (residues 33 to 48 of the predicted gD-1 sequence) was synthesized. This peptide reacted with an anti-gD monoclonal antibody (group VII) previously shown to neutralize the infectivity of HSV-1 and HSV-2. The peptide was also recognized by polyclonal antibodies prepared against purified gD-1 but was less reactive with anti-gD-2 sera. Sera from animals immunized with the synthetic peptide reacted with native gD and neutralized both HSV-1 and HSV-2.     

Control of chicken coccidiosis

Coccidiosis could potentially cause enormous economic loss to the poultry industry, especially in the production of broiler chickens (see Box 1). Losses are currently minimized by chemotherapeutic treatment but the effectiveness of many drugs seems to be declining. In this article, Peter Long and Tom Jeffers discuss the future for coccidial chemotherapy, and the potential for immunological control methods.     

Brain temperature measurements in rats: a comparison of microwave and ambient temperature exposures

In an effort to understand microwave heating better, regional brain and core temperatures of rats exposed to microwave radiation (2450 MHz) or elevated air temperatures were measured in two studies. In general, we have found no substantial evidence for temperature differentials, or "hot spots," in the brain of these animals. In the first study, after a 30-min exposure, no temperature differences between brain regions either after microwave or ambient air exposure were found. However, a highly significant correlation between brain and core temperatures was found and this correlation was the same for both microwave and ambient air heating. In the second study, time-temperature profiles were measured in rats exposed to either 30 mW/cm2 or 36.2 degrees C. In this study, the 30-min exposure period was divided into seven intervals and the change in temperature during each period was analyzed. Only the cortex showed significantly different heating rates between the air heating and microwave heating; however, this difference disappeared after the initial 5 min of exposure.     

Binding of zinc ions to heparin. Analysis by equilibrium dialysis suggests the occurrence of two, entropy-driven, processes.

Analysis by equilibrium dialysis of the binding of Zn2+ to heparin suggested that two interactions, one of high affinity and one of low affinity, occur. The stoichiometry of binding in both cases is about one Zn2+ ion bound per average heparin disaccharide unit. Both types of interaction appear to be entropy-driven.     

Porphyran primary structure. An investigation using beta-agarase I from Pseudomonas atlantica and 13C-NMR spectroscopy

Porphyran, a highly substituted agarose from Porphyra umbilicalis was degraded by highly purified beta-agarase I from Pseudomonas atlantica. This enzyme cleaved at the reducing side of units of beta-neoagarobiose (3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-beta-D-galactopyranose). The oligosaccharides were divided into fractions of low and high molecular weight by dialysis. The permeate (23% of total starting carbohydrate) was separated by ion-exchange into neutral and anionic fractions. Gel filtration of the neutral fraction (19%) resolved two major oligosaccharides. These were shown by 13C-NMR spectroscopy to be 6(3)-O-methyl-neoagarotetraose and 6(3),6(5)-di-O-methyl-neoagarohexaose. Gel filtration of the anionic oligosaccharides (3.3%) revealed two novel monosulphated tetrasaccharides, 6-O-sulphato-alpha-L-galacto-pyranosyl-(1 leads to 3)-beta-D-galactopyranosyl-(1 leads to 4)-3,6-anhydro-alpha-L-galactopyranosyl-(1 leads to 3)-D-galactopyranose and its 6(3)-O-methylated derivative. The 13C-NMR data from the sulphated tetrasaccharides provided a novel reference which was used to characterise higher, partially sulphated fragments in the dialysis permeate. The fraction retained on dialysis (77%) had an average degree of polymerisation of 40 and was homologous with the high-molecular-weight anionic permeate. From 13C-NMR spectroscopy porphyran was found to comprise 49% sulphated disaccharide units and these were calculated to occur in stretches averaging 2.0-2.5 contiguous units.     

Effect of alpha 1-proteinase inhibitor and sulphated polysaccharides on the activity of m beta-acrosin

Purified m beta-acrosin catalysed amidolysis in vitro of several p-nitroanilides with C-terminal arginine residues. alpha 1-proteinase inhibitor inhibited amidolysis catalysed by the enzyme. This effect of alpha 1-Proteinase inhibitor was not prevented by pre-incubation of the enzyme with heparin or any other glycosaminoglycan. Pre-incubation of the enzyme with sulphated dextran or sulphated cellulose alleviated the effect of alpha 1-proteinase inhibitor. These results are discussed in terms of possible in vivo modulation by alpha 1-proteinase inhibitor of acrosin activity.     

Glycopeptides of the Type-Common Glycoprotein gD of Herpes Simplex Virus Types 1 and 2

We have carried out detailed structural studies of the glycopeptides of glycoprotein gD of herpes simplex virus types 1 and 2. We first examined and compared the number of N-asparagine-linked oligosaccharides present in each glycoprotein. We found that treatment of either pgD-1 or pgD-2 with endo-β-N-acetylglucosaminidase H (Endo H) generated three polypeptides which migrated more rapidly than pgD on gradient sodium dodecyl sulfate-polyacrylamide gels. Two of the faster-migrating polypeptides were labeled with [³H]mannose, suggesting that both pgD-1 and pgD-2 contained three N-asparagine-linked oligosaccharides. Second, we characterized the [³H]mannose-labeled tryptic peptides of pgD-1 and pgD-2. We found that both glycoproteins contained three tryptic glycopeptides, termed glycopeptides 1, 2, and 3. Gel filtration studies indicated that the molecular weights of these three peptides were approximately 10,000, 3,900, and 1,800, respectively, for both pgD-1 and pgD-2. Three methods were employed to determine the size of the attached oligosaccharides. First, the [³H]mannose-labeled glycopeptides were treated with Endo H, and the released oligosaccharide was chromatographed on Bio-Gel P6. The size of this molecule was estimated to be approximately 1,200 daltons. Second, Endo H treatment of [³⁵S]methionine-labeled glycopeptide 2 reduced the molecular size of this peptide from approximately 3,900 to approximately 2,400 daltons. Third, glycopeptide 2 isolated from the gD-like molecule formed in the presence of tunicamycin was approximately 2,200 daltons. From these experiments, the size of each N-asparagine-linked oligosaccharide was estimated to be approximately 1,400 to 1,600 daltons. Our experiments indicated that glycopeptides 2 and 3 each contained one N-asparagine-linked oligosaccharide chain. Although glycopeptide 1 was large enough to accommodate more than one oligosaccharide chain, the experiments with Endo H treatment of the glycoprotein indicated that there were only three N-asparagine-linked oligosaccharides present in pgD-1 and pgD-2. Further studies of the tryptic glycopeptides by reverse-phase high-performance liquid chromatography indicated that all of the glycopeptides were hydrophobic in nature. In the case of glycopeptide 2, we observed that when the carbohydrate was not present, the hydrophobicity of the peptide increased. The properties of the tryptic glycopeptides of pgD-1 were compared with the properties predicted from the deduced amino acid sequence of gD-1. The size and amino acid composition compared favorably for glycopeptides 1 and 2. Glycopeptide 3 appeared to be somewhat smaller than would be predicted from the deduced sequence of gD-1. It appears that all three potential glycosylation sites predicted by the amino acid sequence are utilized in gD-1 and that a similar number of glycosylation sites are present in gD-2.     

Release of carboxylating enzymes from maize and sugar cane leaf tissue during progressive grinding

Summary The release of chlorophyll, chloroplasts, o-diphenols, o-diphenol oxidase activity and carboxylating enzyme activity during the grinding of maize and sugar cane leaf tissue has been correlated with the breakage of different types of cell. Enzymes of the photosynthetic carbon reduction cycle were released in the grinding stage during which the bulk of the mesophyll tissue was disrupted and grana-containing chloroplasts released. Since the largest amount of phenol oxidase activity and of phenols was also released at this stage it is likely that the enzymes were partly inhibited by phenol oxidation products and, therefore, underestimated. PEP carboxylase is released earlier in the grinding process. It is concluded that the photosynthetic carbon reduction cycle enzymes studied are located in mesophyll cell chloroplasts and that the PEP carboxylase resides outside the chloroplasts, either in the cytoplasm of mesophyll cells or in colourless tissue. These results are discussed in relation to current theories regarding the assimilation and shuttling of carbon dioxide in leaves of tropical grasses.     

A combination rapid and standard method for identification of Candida albicans

A medium consisting of an aqueous extract of zein, lactose, and Tween 80 is used together with an overlay of 1 % Tween 80 and coverslipping to provide a combined rapid (germ tube) and standard (chlamydospore) method for diagnosis ofCandida albicans. The method is exquisitely sensitive for diagnosis ofC. albicans but lumps chlamydospore-producing strains ofC. tropicalis withC. albicans.