Inhibition of terminal differentiation of B cells mediated by CD27 and CD40 involves signaling through JNK

B cells responding to cognate Ag in vivo undergo clonal expansion that is followed by differentiation into Ab-secreting plasma cells or into quiescent restimulable memory. Both these events occur in the germinal center and require that cells exit from proliferation, but the signals that lead to one or the other of these mutually exclusive differentiation pathways have not been definitively characterized. Previous experiments have shown that signals transduced through the TNFRs CD27 and CD40 at the time of B cell stimulation in vitro or in vivo can influence this cell fate decision by inhibiting terminal differentiation and promoting memory. In this study, we show that the PIQED domain of the cytoplasmic tail of murine CD27 and the adapter molecule TNFR-associated factor 2 are involved in this effect. Using pharmacological inhibitors of signaling intermediates, we identify JNK as being necessary and sufficient for the observed inhibition of terminal differentiation. While JNK is involved downstream of CD40, inhibition of the MEK pathway can also partially restore plasma cell generation, indicating that both signaling intermediates may be involved. We also show that inhibition of induction of IFN regulatory factor 4 and B lymphocyte induced maturation protein 1 are downstream events common to both receptors.     

Loading red blood cells with trehalose: a step towards biostabilization

A method for freeze-drying red blood cells (RBCs) while maintaining a high degree of viability has important implications in blood transfusion and clinical medicine. The disaccharide trehalose, found in animals capable of surviving dehydration can aid in this process. As a first step toward RBC preservation, we present a method for loading RBCs with trehalose. The method is based on the thermal properties of the RBC plasma membranes and provides efficient uptake of the sugar at 37 degrees C in a time span of 7 h. The data show that RBCs can be loaded with trehalose from the extracellular medium through a combination of osmotic imbalance and the phospholipid phase transition, resulting in intracellular trehalose concentrations of about 40 mM. During the loading period, the levels of ATP and 2,3-DPG are maintained close to the levels of fresh RBCs. Increasing the membrane fluidity through the use of a benzyl alcohol results in a higher concentration of intracellular trehalose, suggesting the importance of the membrane physical state for the uptake of the sugar. Osmotic fragility data show that trehalose exerts osmotic protection on RBCs. Flow cytometry data demonstrate that incubation of RBCs in a hypertonic trehalose solution results in a fraction of cells with different complexity and that it can be removed by washing and resuspending the RBCs in an iso-osmotic medium. The data provide an important first step in long-term preservation of RBCs.     

IL-1beta-mediated innate immunity is amplified in the db/db mouse model of type 2 diabetes

Chronic inflammation appears to play a critical role in type 2 diabetes and its complications. Here we tested the hypothesis that this inflammatory dysregulation affects the IL-1beta system and has functional consequences in the brain. Diabetic, db/db, and nondiabetic, db/+, mice were administered i.p. LPS, a potent cytokine inducer, at a dose of 100 microg/kg/mouse. db/db mouse innate immune-associated sickness behavior was 14.8, 33, 44.7, and 34% greater than that of db/+ mice at 2, 4, 8, and 12 h, respectively. When a fixed dose of LPS was used (5 microg/mouse), db/db mouse sickness was again enhanced 18.4, 22.2, and 14.5% at 4, 8, and 12 h as compared with db/+ mice. In diabetic mice, peritoneal macrophages produced more IL-1beta in response to LPS, and peritoneal levels of IL-1beta induced by LPS were increased. Importantly, IL-1R antagonist and type 2 IL-1 receptor (IL-1R2) failed to up-regulate in response to LPS in db/db mice. Finally, both peripheral and central administration of IL-1beta, itself, induced sickness in db/db mice that mimicked the effects of peripheral LPS and was significantly greater than that seen in db/+ mice. Taken together, these results indicate that IL-1beta-mediated innate immunity is augmented in db/db mice both at the periphery and in the brain, and the mechanism is due to diabetes-associated loss of IL-1beta counterregulation.     

Effects of High Temperature Exposure of Spinach Intact Plants and Isolated Thylakoids on Light-Harvesting Complex 2 Protein Phosphorylation

After a 6 min exposure of isolated thylakoids to 43 °C, the extent of phosphorylation of light-harvesting complex of photosystem 2 (LHC2) was higher than in control thylakoids kept at 25 °C. Similarly, the exposure of intact spinach plants to 43 °C in dark for 11 h induced higher extent of thylakoid LHC2 phosphorylation than in control plants kept at 25 °C. The induced ability of LHC2 for enhanced phosphorylation may enable better energy distribution in favour of photosystem 1.     

Influence of cation–π interactions to the structural stability of prokaryotic and eukaryotic translation elongation factors

We have investigated the role of cation–π interactions on translation elongation factors. In our investigation, an average of four significant cation–π interactions were found, that is, an average of one cation–π interaction per 44 residues in the ten elongation factors were observed. The analysis on the influence of short (<±4), medium (>±4 to <±20) and long (>20) range contacts showed that cation–π interactions are mainly formed by medium and long-range contacts. Arg-Tyr pair was found largest in number but energetic contribution of Arg-Trp pair was found most. Preferred secondary structural conformation analysis of the residues involved in cation–π interaction indicates that the cationic Arg prefers to be in helix and Lys having equal probability for helix and strand, whereas the aromatic Phe and Trp were found mostly in helix while Tyr in strand regions. The cation–π interaction residues involved in these proteins were found highly conserved with 48.86% residues having conservation score of ≥6. Analysis of secondary structure preference of the energetically significant cation–π residues in different solvent accessible range indicates that most of the π residues are found buried or partially buried whereas cationic residues were found mostly at the protein surface. The results presented in this study will be useful for structural stability studies in translation elongation factors.     

Diurnal variation in methane efflux at different growth stages of tropical rice

Diurnal variation in CH₄ efflux from continuously flooded fields planted to rice (Oryza sativa L. cv. IR-36) was examined at different crop growth stages using a closed chamber method during the wet season. CH₄ emission showed a distinct diurnal pattern especially at tillering, panicle initiation and maturity stages of a field-grown rice crop, with maximum emission in the early afternoon (12.00 to 15.00) followed by a decline to a minimum around midnight. Among several variables (ambient temperature, flood water temperature, redox potential, soil pH, and root oxidase activity), a significant negative correlation existed between oxidase activity of the root base and diurnal fluctuations in CH₄ efflux at tillering stage. Evidence also suggested that redox status in the rhizosphere region and atmospheric, soil, and water temperatures influenced CH₄ emission from rice fields probably by their contrasting effects on CH₄ production and oxidation.     

Effect of dietary protein and carbohydrate levels on growth,nutrient utilization and body composition in fingerling rohu,Labeo rohita (Hamilton)

Twelve experimental diets (D‐1 to D‐12) in a 4 × 3 factorial design having four protein levels (25, 35, 40 and 45%) and three carbohydrate levels (15, 25 and 35%) were formulated and fed to fingerling rohu, Labeo rohita (5.48 ± 0.02 g) for 60 days in three replicates at 2% BW per day. The best performance of fish in terms of weight gain (%), specific growth rate (SGR; % per day), feed conversion ratio (FCR) and protein efficiency ratio (PER) was recorded with diet D‐9 containing 40% protein and 35% dextrin as a source of dietary carbohydrate. In general, lower protein consumption per kilogram BW was observed at all protein levels with the rise of the dextrin level. The apparent digestibilities of protein and lipid were not affected by the dietary treatments. At the end of the experiment the body composition of animals from all treatments showed lower percentages of moisture and higher percentages of protein as compared to the initial values. A consistent rise in protein retention efficiency was noted in fish fed diets with increasing dextrin levels. The highest protein sparing effect was found in fish fed the diet containing 40% protein and 35% dextrin.