Baculovirus p35 increases pancreatic beta-cell resistance to apoptosis

beta-cells die by apoptosis in type 1 diabetes as a result of autoimmune attack mediated by cytokines, and in type 2 diabetes by various perpetrators including human islet amyloid polypeptide (hIAPP). The cascade of apoptotic events induced by cytokines and hIAPP is mediated through caspases and reactive oxygen species. The baculovirus p35 protein is a potent anti-apoptotic agent shown to be effective in a variety of species and able to inhibit a number of apoptotic pathways. Here, we aimed at determining the protective potential of p35 in beta-cells exposed to cytokines and hIAPP, as well as the effects of p35 on beta-cell function. The p35 gene was introduced into betaTC-tet cells, a differentiated murine beta-cell line capable of undergoing inducible growth-arrest. Both proliferating and growth-arrested cells expressing p35 manifested increased resistance to cytokines and hIAPP, compared with control cells, as judged by cell viability, DNA fragmentation, and caspase-3 activity assays. p35 was significantly more protective in growth-arrested, compared with proliferating, cells. No significant differences were observed in proliferation and insulin content between cells expressing p35 and control cells. In contrast, p35 manifested a perturbing effect on glucose-induced insulin secretion. These findings suggest that p35 could be incorporated as part of a multi-pronged approach of immunoprotective strategies to provide protection from recurring autoimmunity for transplanted beta-cells, as well as in preventive gene therapy in type 1 diabetes. p35 may also be protective from beta-cell damage caused by hIAPP in type 2 diabetes.     

Receptor editing in positive and negative selection of B lymphopoiesis

In B lymphopoiesis, Ag receptor expression and signaling are critical to determine developmental progression, survival, and activation. Several positive and negative selection checkpoints to test this receptor have been described in B lymphopoiesis, aiming to ensure the generation of functionally competent, nonautoimmune repertoire. Secondary Ag receptor gene recombination allows B lymphocytes to replace an inappropriate receptor with a new receptor, a mechanism called receptor editing. This salvage mechanism uncouples the Ag receptor fate from that of the cell itself, suggesting that B cell repertoire is regulated by a process of receptor selection. Secondary rearrangements are stimulated in different stages of B cell development, where editing of the receptor is necessary to fulfill stage-specific requirements. In this study, we discuss the contribution of receptor editing in B lymphopoiesis and its regulation by positive and negative selection signals.     

Regulation of collagen deposition and lysyl oxidase by tumor necrosis factor-alpha in osteoblasts

Tumor necrosis factor-alpha (TNF-alpha) inhibits osteoblast function in vitro by inhibiting collagen deposition. Studies generally support that TNF-alpha does not inhibit collagen biosynthesis by osteoblasts but that collagen deposition is in some way diminished. The study investigated TNF-alpha regulation of biosynthetic enzymes and proteins crucial for posttranslational extracellular collagen maturation in osteoblasts including procollagen C-proteinases, procollagen C-proteinase enhancer, and lysyl oxidase. The working hypothesis is that such regulation could inhibit collagen deposition by osteoblasts. We report that in phenotypically normal MC3T3-E1 osteoblasts, TNF-alpha decreases collagen deposition without decreasing collagen mRNA levels or procollagen protein synthesis. Analyses of the cell layers revealed that TNF-alpha diminished the levels of mature collagen cross-links, pyridinoline and deoxypyridinoline. Further analyses revealed that the mRNA expression for lysyl oxidase, the determining enzyme required for collagen cross-linking, is down-regulated by TNF-alpha in a concentration- and time-dependent manner by up to 50%. The decrease was accompanied by a significant reduction of lysyl oxidase protein levels and enzyme activity. By contrast, Northern and Western blotting studies revealed that procollagen C-proteinases bone morphogenic protein-1 and mammalians Tolloid and procollagen C-proteinase enhancer were expressed in MC3T3-E1 cells and not down-regulated. The data together demonstrate that TNF-alpha does not inhibit collagen synthesis but does inhibit the expression and activity of lysyl oxidase in osteoblasts, thereby contributing to perturbed collagen cross-linking and accumulation. These studies identify a novel mechanism in which proinflammatory cytokine modulation of an extracellular biosynthetic enzyme plays a determining role in the control of collagen accumulation by osteoblasts.     

Comparison of the Antimicrobial Adhesion Potential of Human Body Fluid Glycoconjugates Using Fucose-Binding Lectin (PA-IIL) of Pseudomonas aeruginosa and Ulex europaeus Lectin (UEA-I)

Pseudomonas aeruginosa produces a fucose-binding lectin (PA-IIL) which strongly binds to human cells. This lectin was shown to be highly sensitive to inhibition by fucose-bearing human milk glycoproteins. Since the glycans of these glycoproteins mimic human cell receptors, they may function as decoys in blocking lectin-dependent pathogen adhesion to the host cells. Human saliva and seminal fluid also contain such compounds, and body fluids of individuals who are “secretors” express additional fucosylated (alpha 1,2) residues. The latter are selectively detected by Ulex europaeus lectin UEA-I. The aim of the present research was to compare the PA-IIL and UEA-I interactions with human salivas and seminal fluids of “secretors” and “nonsecretors” with those obtained with the respective milks. Using hemagglutination inhibition and Western blot analyses, we showed that PA-IIL interactions with the saliva and seminal fluid glycoproteins were somewhat weaker than those obtained with the milk and that “nonsecretor” body fluids were not less efficient than those of “secretors” in PA-IIL blocking. UEA-I, which interacted only with the “secretors” glycoproteins, was most sensitive to those of the seminal fluids.     

Various competitive interactions explain niche separation in crop‐dwelling web spiders

Competition for resources is a major organizing principle in communities of organisms that share similar ecological niches. Niche separation by means of exploitation or interference competition was investigated in two taxa of crop‐inhabiting spiders that overlap in microhabitat use and have similar web design. Competition for prey and web sites was tested in microcosm experiments with the most common species that build sheet‐webs: Enoplognatha gemina (Theridiidae) and Alioranus pastoralis (Linyphiidae). A field survey over the crop season provided data on spatial and temporal dispersion of Enoplognatha spp. (Theridiidae) and linyphiid spiders (Linyphiidae) and on availability of prey over the season. In the microcosm experiments, both taxa took springtails as prey, but only Enoplognatha fed on aphids. Differences in diet, however, could not be attributed to either exploitative or interference competition. Spatial separation of websites was attained by vertical displacement of webs in the vegetation (Enoplognatha) and by avoidance of patches occupied by conspecific or heterospecific individuals (linyphiids). In the field, densities of linyphiids and Enoplognatha were correlated negatively and webs were over‐dispersed relative to a random distribution. Both taxa colonized the field at the start of the season; linyphiids colonized as adults, quickly reproduced, and had a second adult peak; Enoplognatha matured in the middle of the season and their numbers remained fairly constant over the season. The combined experimental manipulations and field data suggest that niche separation occurs at different scales. The hypothesis of competition for websites was partially supported, while prey preference (or tolerance) and temporal differences in life history stages also may explain the negative correlations between densities of the two taxa.     

Does saline water consumption affect feeding and fuel deposition rate of a staging,long‐distance migrating passerine?

To accomplish their enduring journeys, migrating birds accumulate fuel consisting mainly of lipids in stopover sites located throughout their migration routes. Fuel deposition rate (FDR) is considered a key parameter determining the speed of migration and thereby bird fitness, and recent studies have demonstrated the positive effects of fresh water consumption on the FDR of migrating blackcaps Sylvia atricapilla. Sewage water reservoirs, characterized by higher water salinity than fresh water, were extensively built in different parts of the world and are used by birds during their travel, but their effects on wildlife and specifically on migrating birds have been largely overlooked thus far. We experimentally examined the effects of water salinity on blackcap FDR during migration. We captured birds in an autumn stopover site, transported them to the laboratory and provided them with fruits, mealworms and water of different salinity levels (0.3, 4.5 and 9‰ NaCl) for several days. We examined the effects of water salinity on the blackcaps’ diet, water consumption and FDR and found that FDR was mainly affected by fruit consumption rate and not by the water salinity levels. Water salinity nevertheless caused elevated water consumption as the birds consumed almost 3 times more saline water than fresh water per consumed fruit mass. Our work is the first to explore the consequences of saline water consumption on migrating passerines, specifically suggesting that anthropogenic alterations of habitats by sewage water treatment facilities may modulate bird nutrition and diet.