Bcl-2 allows effector and memory CD8+ T cells to tolerate higher expression of Bim

As acute infections resolve, most effector CD8(+) T cells die, whereas some persist and become memory T cells. Recent work showed that subsets of effector CD8(+) T cells, identified by reciprocal expression of killer cell lectin-like receptor G1 (KLRG1) and CD127, have different lifespans. Similar to previous reports, we found that effector CD8(+) T cells reported to have a longer lifespan (i.e., KLRG1(low)CD127(high)) have increased levels of Bcl-2 compared with their shorter-lived KLRG1(high)CD127(low) counterparts. Surprisingly, we found that these effector KLRG1(low)CD127(high) CD8(+) T cells also had increased levels of Bim compared with KLRG1(high)CD127(low) cells. Similar effects were observed in memory cells, in which CD8(+) central memory T cells expressed higher levels of Bim and Bcl-2 than did CD8(+) effector memory T cells. Using both pharmacologic and genetic approaches, we found that survival of both subsets of effector and memory CD8(+) T cells required Bcl-2 to combat the proapoptotic activity of Bim. Interestingly, inhibition or absence of Bcl-2 led to significantly decreased expression of Bim in surviving effector and memory T cells. In addition, manipulation of Bcl-2 levels by IL-7 or IL-15 also affected expression of Bim in effector CD8(+) T cells. Finally, we found that Bim levels were significantly increased in effector CD8(+) T cells lacking Bax and Bak. Together, these data indicate that cells having the highest levels of Bim are selected against during contraction of the response and that Bcl-2 determines the level of Bim that effector and memory T cells can tolerate.     

A major role for Bim in regulatory T cell homeostasis

We have previously shown that regulatory T cells (Treg) accumulate dramatically in aged animals and negatively impact the ability to control persistent infection. However, the mechanisms underlying the age-dependent accrual of Treg remain unclear. In this study, we show that Treg accumulation with age is progressive and likely not the result of increased thymic output, increased peripheral proliferation, or from enhanced peripheral conversion. Instead, we found that Treg from aged mice are more resistant to apoptosis than Treg from young mice. Although Treg from aged mice had increased expression of functional IL-7Rα, we found that IL-7R signaling was not required for maintenance of Treg in vivo. Notably, aged Treg exhibit decreased expression of the proapoptotic molecule Bim compared with Treg from young mice. Furthermore, in the absence of Bim, Treg accumulate rapidly, accounting for >25% of the CD4(+) T cell compartment by 6 mo of age. Additionally, accumulation of Treg in Bim-deficient mice occurred after the cells left the transitional recent thymic emigrant compartment. Mechanistically, we show that IL-2 drives preferential proliferation and accumulation of Bim(lo) Treg. Collectively, our data suggest that chronic stimulation by IL-2 leads to preferential expansion of Treg having low expression of Bim, which favors their survival and accumulation in aged hosts.     

A mathematical model for the peristaltic flow of chyme movement in small intestine

A mathematical model based on viscoelastic fluid (fractional Oldroyd-B model) flow is considered for the peristaltic flow of chyme in small intestine, which is assumed to be in the form of an inclined cylindrical tube. The peristaltic flow of chyme is modeled more realistically by assuming that the peristaltic rush wave is a sinusoidal wave, which propagates along the tube. The governing equations are simplified by making the assumptions of long wavelength and low Reynolds number. Analytical approximate solutions of problem are obtained by using homotopy analysis method and convergence of the obtained series solution is properly checked. For the realistic values of the emerging parameters such as fractional parameters, relaxation time, retardation time, Reynolds number, Froude number and inclination of tube, the numerical results for the pressure difference and the frictional force across one wavelength are computed and discussed the roles played by these parameters during the peristaltic flow. On the basis of this study, it is found that the first fractional parameter, relaxation time and Froude number resist the movement of chyme, while, the second fractional parameter, retardation time, Reynolds number and inclination of tube favour the movement of chyme through the small intestine during pumping. It is further revealed that size of trapped bolus reduces with increasing the amplitude ratio whereas it is unaltered with other parameters.     

Clarithromycin, a cytochrome P450 inhibitor, can reverse mefloquine resistance in Plasmodium yoelii nigeriensis- infected Swiss mice

During the last 2 decades there have been numerous reports of the emergence of mefloquine resistance in Southeast Asia and nearly 50% resistance is reported in Thailand. A World Health Organization report (2001) considers mefloquine as an important component of ACT (artesunate+mefloquine) which is the first line of treatment for the control of uncomplicated/multi-drug resistant (MDR) Plasmodium falciparum malaria. In view of the emergence of resistance towards this drug, it is proposed to develop new drug combinations to prolong the protective life of this drug. Prior studies have suggested that mefloquine resistance can be overcome by a variety of agents such as ketoconazole, cyproheptadine, penfluridol, Icajine and NP30. The present investigation reports that clarithromycin (CLTR), a new macrolide, being a potent inhibitor of Cyt. P450 3A4, can exert significant resistance reversal action against mefloquine resistance of plasmodia. Experiments were carried out to find out the curative dose of CLTR against multi-drug resistant P. yoelii nigeriensis. Mefloquine (MFQ) and clarithromycin (CLTR) combinations have been used for the treatment of this MDR parasite. Different dose combinations of these two drugs were given to the infected mice on day 0 (prophylactic) and day 1 with established infection (therapeutic) to see the combined effect of these combinations against the MDR malaria infection. With a dose of 32 mg/kg MFQ and 225 mg/kg CLTR, 100% cure was observed, while in single drug groups, treated with MFQ or CLTR, the cure was zero and 40% respectively. Therapeutically, MFQ and CLTR combinations 32+300 mg/kg doses cleared the established parasitaemia on day 10. Single treatment with MFQ or CLTR showed considerable suppression of parasitaemia on day 14 but neither was curative. Follow-up of therapeutically treated mice showed enhanced anti-malarial action as reflected by their 100% clearance of parasitaemia. The present study reveals that CLTR is a useful antibiotic to be used as companion drug with mefloquine in order to overcome mefloquine resistance in plasmodia.     

Influence of Exogenous Silicon Addition on Aluminium Tolerance in Rice Seedlings

For better understanding of silicon (Si)-mediated increase in metal tolerance in plants, the alterations in growth, mineral elements and anatomical features were investigated in rice seedlings exposed to Si (10?μM) and aluminium (Al; 50?μM). Al decreased seedling growth which was accompanied by an increase in Al accumulation. In contrast, addition of Si alleviated toxic effects of Al and led to the decrease in Al accumulation. Magnesium (Mg) and zinc (Zn) contents were decreased by Al; however, Si addition prevented decrease in Mg and Zn contents. Manganese (Mn) content was increased by Al while Si addition did not have a significant effect on Mn content under Al treatment. Al exposure decreased frequency of stomata and root hairs, length of root hairs and leaf epidermal cells, and distorted the structure and integrity of mesophyll cells and phloem; however, addition of Si reduced these abnormalities. Results showed that Si addition protected rice seedlings against Al toxicity by decreasing Al accumulation and by maintaining level of some mineral elements, and the key structures of leaf and root.     

Studies on non-symbiotic diazotrophic bacterial populations of coastal arable saline soils of India

The effect of fluctuations of salinity in three different seasons on diazotrophic populations and N₂ fixation in six mono cropped rice field soils of the coastal region of the Gangetic delta of West Bengal, India, was studied. The average pH, ECe, organic carbon and total nitrogen of the soils ranged from 4.99–7.08, 2.02–19.58 dSm⁻¹, 4.68–12.03 g kg⁻¹ and 0.44–1.70 g kg ⁻¹, respectively. The average log colony forming units of the bacterial populations and N₂-fixation in the soils varied from 4.61 to 5.86 and 2.74 to 4.52 mg N₂ fixed 50 ml ⁻¹ culture media respectively, with the lowest value recorded in summer. Recovery of microorganisms and N₂- fixation gradually decreased with extraneous addition of NaCl in the culture media. All the eight isolates were Gram positive, spore and capsule formers. They could utilize glucose, sucrose, mannitol, starch, citrate and nitrate, and were catalase and gelatinase positive, but indole, methyl red and Vogues Proskauer reaction negative. The organisms produced alkaline reaction on TSI agar slant. The acetylene reduction assay of the isolates at 0 and 1% NaCl in the culture media were 4.51–164.52 and 1.72–100.6 nmole C₂H₄ ml⁻¹ culture media in 72 h, respectively. The isolates could fix 2.42–4.45 and 2.04–4.08 mg N₂ fixed 50 ml⁻¹ culture media at 0 and 1% NaCl in the culture media respectively. 16S rDNA sequences of the isolates were similar to the species: Bacillus sp. isolate 28A, Bacillus sp. MOLA 87, Bacillus sp. By113 (B)Ydz-dh, Bacillus sp. PN13, Bacillus licheniformis strain RH101, Bacterium Antarctica 14, Bacillus sp. PN13 and Bacillus megaterium.     

An alkaline phosphatase/phosphodiesterase, PhoD, induced by salt stress and secreted out of the cells of Aphanothece halophytica, a halotolerant cyanobacterium

Alkaline phosphatases (APases) are important enzymes in organophosphate utilization. Three prokaryotic APase gene families, PhoA, PhoX, and PhoD, are known; however, their functional characterization in cyanobacteria largely remains to be clarified. In this study, we cloned the phoD gene from a halotolerant cyanobacterium, Aphanothece halophytica (phoD(Ap)). The deduced protein, PhoD(Ap), contains Tat consensus motifs and a peptidase cleavage site at the N terminus. The PhoD(Ap) enzyme was activated by Ca(2+) and exhibited APase and phosphodiesterase (APDase) activities. Subcellular localization experiments revealed the secretion and processing of PhoD(Ap) in a transformed cyanobacterium. Expression of the phoD(Ap) gene in A. halophytica cells was upregulated not only by phosphorus (P) starvation but also under salt stress conditions. Our results suggest that A. halophytica cells possess a PhoD that participates in the assimilation of P under salinity stress.     

Genome sequence of B. amyloliquefaciens type strain DSM7(T) reveals differences to plant-associated B. amyloliquefaciens FZB42

The complete genome sequence of Bacillus amyloliquefaciens type strain DSM7^T is presented. A comparative analysis between the genome sequences of the plant associated strain FZB42 (Chen et al., 2007) with the genome of B. amyloliquefaciens DSM7^T revealed obvious differences in the variable part of the genomes, whilst the core genomes were found to be very similar. The strains FZB42 and DSM7^T have in common 3345 genes (CDS) in their core genomes; whilst 547 and 344 CDS were found to be unique in DSM7^T and FZB42, respectively. The core genome shared by both strains exhibited 97.89% identity on amino acid level. The number of genes representing the core genome of the strains FZB42, DSM7^T, and Bacillus subtilis DSM10^T was calculated as being 3098 and their identity was 92.25%. The 3,980,199 bp genome of DSM7^T contains numerous genomic islands (GI) detected by different methods. Many of them were located in vicinity of tRNA, glnA, and glmS gene copies. In contrast to FZB42, but similar to B. subtilis DSM10^T, the GI were enriched in prophage sequences and often harbored transposases, integrases and recombinases. Compared to FZB42, B. amyloliquefaciens DSM7^T possessed a reduced potential to non-ribosomally synthesize secondary metabolites with antibacterial and/or antifungal action. B. amyloliquefaciens DSM7^T did not produce the polyketides difficidin and macrolactin and was impaired in its ability to produce lipopeptides other than surfactin. Differences established within the variable part of the genomes, justify our proposal to discriminate the plant-associated ecotype represented by FZB42 from the group of type strain related B. amyloliquefaciens soil bacteria.     

Prospective isolation of adult neural stem cells from the mouse subependymal zone

Neural stem cells (NSCs) have the remarkable capacity to self-renew and the lifelong ability to generate neurons in the adult mammalian brain. However, the molecular and cellular mechanisms contributing to these behaviors are still not understood. Now that prospective isolation of the NSCs has become feasible, these mechanisms can be studied. Here we describe a protocol for the efficient isolation of adult NSCs, by the application of a dual-labeling strategy on the basis of their glial identity and ciliated nature. The cells are isolated from the lateral ventricular subependymal zone (SEZ) of adult hGFAP-eGFP (human glial fibrillary acidic protein-enhanced green fluorescent protein) transgenic mice by fluorescence-activated cell sorting. Staining against prominin1 (CD133) allows the isolation of the NSCs (hGFAP-eGFP(+)/prominin1(+)), which can be further subdivided by labeling with the fluorescent epidermal growth factor. This protocol, which can be completed in 7 h, allows the assessment of quantitative changes in SEZ NSCs and the examination of their molecular and functional characteristics.     

Monocrotophos induced apoptosis in PC12 cells: role of xenobiotic metabolizing cytochrome P450s

Monocrotophos (MCP) is a widely used organophosphate (OP) pesticide. We studied apoptotic changes and their correlation with expression of selected cytochrome P450s (CYPs) in PC12 cells exposed to MCP. A significant induction in reactive oxygen species (ROS) and decrease in glutathione (GSH) levels were observed in cells exposed to MCP. Following the exposure of PC12 cells to MCP (10(-5) M), the levels of protein and mRNA expressions of caspase-3/9, Bax, Bcl(2), P(53), P(21), GSTP1-1 were significantly upregulated, whereas the levels of Bclw, Mcl1 were downregulated. A significant induction in the expression of CYP1A1/1A2, 2B1/2B2, 2E1 was also observed in PC12 cells exposed to MCP (10(-5) M), whereas induction of CYPs was insignificant in cells exposed to 10(-6) M concentration of MCP. We believe that this is the first report showing altered expressions of selected CYPs in MCP-induced apoptosis in PC12 cells. These apoptotic changes were mitochondria mediated and regulated by caspase cascade. Our data confirm the involvement of specific CYPs in MCP-induced apoptosis in PC12 cells and also identifies possible cellular and molecular mechanisms of organophosphate pesticide-induced apoptosis in neuronal cells.