The viral sigma1 protein and glycoconjugates containing alpha2-3-linked sialic acid are involved in type 1 reovirus adherence to M cell apical surfaces

Type 1 reoviruses invade the intestinal mucosa of mice by adhering selectively to M cells in the follicle-associated epithelium and then exploiting M cell transport activity. The purpose of this study was to identify the apical cell membrane component and viral protein that mediate the M cell adherence of these viruses. Virions and infectious subviral particles of reovirus type 1 Lang (T1L) adhered to rabbit M cells in Peyer's patch mucosal explants and to tissue sections in an overlay assay. Viral adherence was abolished by pretreatment of sections with periodate and in the presence of excess sialic acid or lectins MAL-I and MAL-II (which recognize complex oligosaccharides containing sialic acid linked alpha2-3 to galactose). The binding of T1L particles to polarized human intestinal (Caco-2(BBe)) cell monolayers was correlated with the presence of MAL-I and MAL-II binding sites, blocked by excess MAL-I and -II, and abolished by neuraminidase treatment. Other type 1 reovirus isolates exhibited MAL-II-sensitive binding to rabbit M cells and polarized Caco-2(BBe) cells, but type 2 or type 3 isolates including type 3 Dearing (T3D) did not. In assays using T1L-T3D reassortants and recoated viral cores containing T1L, T3D, or no sigma1 protein, MAL-II-sensitive binding to rabbit M cells and polarized Caco-2(BBe) cells was consistently associated with the T1L sigma1. MAL-II-recognized oligosaccharide epitopes are not restricted to M cells in vivo, but MAL-II immobilized on virus-sized microparticles bound only to the follicle-associated epithelium and M cells. The results suggest that selective binding of type 1 reoviruses to M cells in vivo involves interaction of the type 1 sigma1 protein with glycoconjugates containing alpha2-3-linked sialic acid that are accessible to viral particles only on M cell apical surfaces.     

Studies on thermal degradation and termite resistant properties of chemically modified wood

A series of experiments were carried out to examine the resistant capacity of a chemically treated hard wood, Anthocephalus cadamba (Roxb) Miq. to thermal and termite degradation. The treatment with thermosetting resins viz. urea formaldehyde (UF), melamine formaldehyde (MF) and phenol formaldehyde (PF) at 31-33 levels of weight percent gain (WPG) increased the strength property i.e. modulus of rupture (MOR) by 7.50-21.02% and stiffness i.e. modulus of elasticity (MOE) by 9.50-12.18% over the untreated one with no remarkable effect on specific gravity. The treated samples were found resistant to termite attack, while the untreated one was badly damaged by termites on 12 months' exposure to a termite colony. The thermal degradations of untreated and treated wood samples were studied using thermogravimetric (TGA) and differential thermogravimetric (DTG) techniques at heating rates 20 and 30 degrees C min(-1) in temperature range 30-650 degrees C. The treated wood was found to be thermally more stable than the untreated one.     

Stereochemistry of strictic acid and related furano-diterpenes from conyza japonica and grangea maderaspatana

Extraction of Conyza japonica gave strictic acid, ent-2β-hydroxy-15,16-epoxy-3,13(16),14-clerodatrien-18-oic acid and 5,7-dihydroxy-3,8,4′-trimethoxyflavone. Extraction of Grangea maderaspatana gave (-)-hardwickiic acid, ent-15,16-epoxy-1,3,13(16),14-clerodatetraen-18-oic acid and 3-hydroxy-8-acetoxypentadeca-1,9,14-trien-4,6-diyne. The structure of ent-2β-hydroxy-15,16-epoxy-3,13(16),14-cleroclatrien-18-oic acid was deduced by spectroscopic methods and by partial synthesis from (-)-hardwickiic acid and the stereochemistries of strictic acid and (ent-15,16-epoxy-1,3,13(16),14-clerodatraen-18-oic acid were established by correlation with ent-2β-hydroxy-15,16-epoxy-3,13(16),14-clerodatrien-18-oic acid.     

Presynaptic modulation of amino acid release from synaptosomes

Using synaptosomes prepared from whole rat brain, the spontaneous, calcium-independent, and calcium-dependent release of glutamate and GABA was assessed. Time intervals of 1–30 seconds were studied. Spontaneous release of glutamate (but not GABA) was elevated by 10 M NMDA or AMPA by thirty seconds. This stimulation was partially calcium-dependent. Calcium-dependent release induced by 30 mM KCl was biphasic, confirming previous findings. This release was stimulated at all time periods by the presence of 10 M NMDA or AMPA in an antagonist-sensitive manner. These data suggest that glutamate and GABA are released from vesicular stores in rat synaptosomes and that some of this release is modulated by presynaptic glutamate receptors.     

Characterization of Plasma membrane of winter wheat (TritiCum aestivum L.) Seedlings and Changes in ATP hydrolysing activity under low temparature stress

Plasma membranes were isolated from young seedlings of cold hardened and non-hardened winter wheat using aqueous polymer two phase system consisting of dextran-polyethyleneglycol. Electron microscopic analysis and activities of various marker enzymes revealed that the upper phase was enriched in plasma membrane and free from contamination by other membranes. The fraction thus obtained possessed Mg ATP hydrolysing activity with an optimum pH 6.5 and the activity was stimulated by K⁺. The activity was highly sensitive to orthovanadate, diethylstilbesterol, dicyclohexylcarbodimide, insensitive to azide and nitrate. It was suggested that the ATP hydrolysis was catalysed by H⁺ ATPase. Mg ATP hydrolysing activity in cold hardened seedlings was 38% higher than that from non-hardened seedlings.     

Genetic control of retinal specification and determination in Drosophila

The Drosophila compound eye has long served as an outstanding model system to study many processes, including cell fate specification, cell division, cell growth and cell death. In addition, exploring the molecular basis of eye specification in Drosophila has identified a set of nuclear factors that trigger the conversion of a group of multipotent epithelial cells into eye primordia. These nuclear factors act in complex networks to regulate retinal specification and appear to be conserved throughout phylogeny. Finally, evidence suggests that these nuclear networks have been co-opted to specify cell fates in other tissues. We review the latest developments in the field of retinal specification in Drosophila and discuss several future directions that remain open for investigation.     

Small mammal trapping in tropical montane forests of the Upper Nilgiris, southern India: An evaluation of capture-recapture models in estimating abundance

Capture-mark-recapture was used to study small mammal populations in tropical montane forests in southern India. Eleven plots in six montane forest patches were sampled from February-October, 1994. Six species were captured, including four rodents and two shrews. PROGRAM CAPTURE was used to derive estimates of density of the most abundant species in the study area,Rattus rattus Linnaeus. The coefficient of variation of the density estimate was used as an index of precision. The coefficient of variation decreased exponentially with increasing capture probability and with an increase in trapping duration. The coefficient of variation and the capture probability were not correlated with estimates of density. The density estimate increased with trapping duration, as did trap mortality. The latter may have been due to the trend of increased mortality with recaptures of the same individual, which in turn may have been due to weight loss over consecutive captures. Estimates of density derived using four estimators were different for 2, 3, 4 and 5 days of trapping. The coefficient of variation was highest for the generalized removal estimate and lowest for Darroch’s estimate. The models and estimators could not be applied to more than one species, and for this species, only in select habitats in a few seasons. Therefore, models of density estimation developed for temperate areas may not be suitable for tropical habitats due to low densities of small mammals in these habitats.     

Putative autocleavage of reovirus mu1 protein in concert with outer-capsid disassembly and activation for membrane permeabilization

Capsid proteins of several different families of non-enveloped animal viruses with single-stranded RNA genomes undergo autocatalytic cleavage (autocleavage) as a maturation step in assembly. Similarly, the 76 kDa major outer-capsid protein mu1 of mammalian orthoreoviruses (reoviruses), which are non-enveloped and have double-stranded RNA genomes, undergoes putative autocleavage between residues 42 and 43, yielding N-terminal N-myristoylated fragment mu1N and C-terminal fragment mu1C. Cleavage at this site allows release of mu1N, which is thought to be critical for penetration of the host-cell membrane during cell entry. Most previous studies have suggested that cleavage at the mu1N/mu1C junction precedes addition to the outer capsid during virion assembly, such that only a small number of the mu1 subunits in mature virions remain uncleaved at that site (approximately 5%). In this study, we varied the conditions for disruption of virions before running the proteins on denaturing gels and in several circumstances recovered much higher levels of uncleaved mu1 (up to approximately 60%). Elements of the disruption conditions that allowed greater recovery of uncleaved protein were increased pH, absence of reducing agent, and decreased temperature. These same elements allowed comparably higher levels of the mu1delta protein, in which cleavage at the mu1N/delta junction has not occurred, to be recovered from particle uncoating intermediates in which mu1 had been previously cleaved by chymotrypsin in a distinct protease-sensitive region near residue 580. The capacity to recover higher levels of mu1delta following disruption of these particles for electrophoresis was lost, however, in concert with a series of structural changes that activate the particles for membrane permeabilization, suggesting that the putative autocleavage is itself one of these changes.