The Bicarbonate Transporter Is Essential for Bacillus anthracis Lethality

In the pathogenic bacterium Bacillus anthracis, virulence requires induced expression of the anthrax toxin and capsule genes. Elevated CO₂/bicarbonate levels, an indicator of the host environment, provide a signal ex vivo to increase expression of virulence factors, but the mechanism underlying induction and its relevance in vivo are unknown. We identified a previously uncharacterized ABC transporter (BAS2714-12) similar to bicarbonate transporters in photosynthetic cyanobacteria, which is essential to the bicarbonate induction of virulence gene expression. Deletion of the genes for the transporter abolished induction of toxin gene expression and strongly decreased the rate of bicarbonate uptake ex vivo, demonstrating that the BAS2714-12 locus encodes a bicarbonate ABC transporter. The bicarbonate transporter deletion strain was avirulent in the A/J mouse model of infection. Carbonic anhydrase inhibitors, which prevent the interconversion of CO₂ and bicarbonate, significantly affected toxin expression only in the absence of bicarbonate or the bicarbonate transporter, suggesting that carbonic anhydrase activity is not essential to virulence factor induction and that bicarbonate, and not CO₂, is the signal essential for virulence induction. The identification of this novel bicarbonate transporter essential to virulence of B. anthracis may be of relevance to other pathogens, such as Streptococcus pyogenes, Escherichia coli, Borrelia burgdorferi, and Vibrio cholera that regulate virulence factor expression in response to CO₂/bicarbonate, and suggests it may be a target for antibacterial intervention.     

Biological activity and the Earth's surface evolution: Insights from carbon,sulfur, nitrogen and iron stable isotopes in the rock record

The search for early Earth biological activity is hindered by the scarcity of the rock record. The very few exposed sedimentary rocks have all been affected by secondary processes such as metamorphism and weathering, which might have distorted morphological microfossils and biogenic minerals beyond recognition and have altered organic matter to kerogen. The search for biological activity in such rocks therefore relies entirely on chemical, molecular or isotopic indicators. A powerful tool used for this purpose is the stable isotope signature of elements related to life (C, N, S, Fe). It provides key informations not only on the metabolic pathways operating at the time of the sediment deposition, but more globally on the biogeochemical cycling of these elements and thus on the Earth's surface evolution. Here, we review the basis of stable isotope biogeochemistry for these isotopic systems. Rather than an exhaustive approach, we address some examples to illustrate how they can be used as biosignatures of early life and as proxies for its environment, while keeping in mind what their limitations are. We then focus on the covariations among these isotopic systems during the Archean time period to show that they convey important information both on the evolution of the redox state of the terrestrial surface reservoirs and on co-occurring ecosystems in the Archean.     

Interspecies variation of Kitasatospora recifensis endophytic from yam bean producing thermostable amylases in alternative media

An endophytic actinomycete isolated from tubers of yam beam (Pachyrhizus erosus L. Urban) was classified as a novel species nominated Kitasatospora recifensis based in phenotypic and genotypic analysis (16S rDNA gene sequence). Monosporic culture using specific ISP2 media revealed three interspecies, which were identified by DNA southern hybridization (Wild strain 13817 W, Aerial Mycelium strain 13817 AM and Vegetative Mycelium strain 13817 VM). The strains were tested for the production of amylolitic enzymes in alternative media. Maximum yields for both enzymes were observed in starch-casein. Higher α-amylase was obtained with strain 13817 W in starch-urea, and amyloglucosidase with strain 13817 AM in starch-ammonium that are economic sources and may be important for industrial purposes. Type strain (DAUFPE 13817 ^T  = KCTC 9972^T = DSM 44943^T).     

Arp2/3-dependent mechanical control of morphogenetic robustness in an inherently challenging environment

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Targeting sphingosine kinase 1 (SK1) enhances oncogene-induced senescence through ceramide synthase 2 (CerS2)-mediated generation of very-long-chain ceramides

Senescence is an antiproliferative mechanism that can suppress tumor development and can be induced by oncogenes such as genes of the Ras family. Although studies have implicated bioactive sphingolipids (SL) in senescence, the specific mechanisms remain unclear. Here, using MCF10A mammary epithelial cells, we demonstrate that oncogenic K-Ras (Kirsten rat sarcoma viral oncogene homolog) is sufficient to induce cell transformation as well as cell senescence—as revealed by increases in the percentage of cells in the G1 phase of the cell cycle, p21^{WAF1/Cip1/CDKN1A} (p21) expression, and senescence-associated β-galactosidase activity (SA-β-gal). Furthermore, oncogenic K-Ras altered SL metabolism, with an increase of long-chain (LC) C18, C20 ceramides (Cer), and very-long-chain (VLC) C22:1, C24 Cer, and an increase of sphingosine kinase 1 (SK1) expression. Since Cer and sphingosine-1-phosphate have been shown to exert opposite effects on cellular senescence, we hypothesized that targeting SK1 could enhance oncogenic K-Ras-induced senescence. Indeed, SK1 downregulation or inhibition enhanced p21 expression and SA-β-gal in cells expressing oncogenic K-Ras and impeded cell growth. Moreover, SK1 knockdown further increased LC and VLC Cer species (C18, C20, C22:1, C24, C24:1, C26:1), especially the ones increased by oncogenic K-Ras. Fumonisin B1 (FB1), an inhibitor of ceramide synthases (CerS), reduced p21 expression induced by oncogenic K-Ras both with and without SK1 knockdown. Functionally, FB1 reversed the growth defect induced by oncogenic K-Ras, confirming the importance of Cer generation in the senescent phenotype. More specifically, downregulation of CerS2 by siRNA blocked the increase of VLC Cer (C24, C24:1, and C26:1) induced by SK1 knockdown and phenocopied the effects of FB1 on p21 expression. Taken together, these data show that targeting SK1 is a potential therapeutic strategy in cancer, enhancing oncogene-induced senescence through an increase of VLC Cer downstream of CerS2.Subject terms: Cancer metabolism, Senescence     

The extended Kalman filter applied to a continuous culture model

The temperature-dependent endogenous metabolism model of single species continuous culture dynamics is utilized in the computer simulation of the Kalman filter state estimation technique. Parameters of the nonlinear equations can be “tracked” while variance in measured states can be damped. The state estimator is illustrated in, the context of conventional control strategies.     

Performance and molecular evaluation of an anaerobic system with suspended biomass for treating wastewater with high fat content after enzymatic hydrolysis

The effect of a lipase-rich fungal enzymatic preparation, produced by a Penicillium sp. during solid-state fermentation, was evaluated in an anaerobic digester treating dairy wastewater with 1200 mg of oil and grease/L. The oil and grease hydrolysis step was carried out with 0.1% (w/v) of solid enzymatic preparation at 30 °C for 24 h, and resulted in a final free acid concentration eight times higher than the initial value. The digester operated in sequential batches of 48 h at 30 °C for 245 days, and had high chemical oxygen demand (COD) removal efficiencies (around 90%) when fed with pre-hydrolyzed wastewater. However, when the pre-hydrolysis step was removed, the anaerobic digester performed poorly (with an average COD removal of 32%), as the oil and grease accumulated in the biomass and effluent oil and grease concentration increased throughout the operational period. PCR-DGGE analysis of the Bacteria and Archaea domains revealed remarkable differences in the microbial profiles in trials conducted with and without the pre-hydrolysis step, indicating that differences observed in overall parameters were intrinsically related to the microbial diversity of the anaerobic sludge.     

Evidence for a polyamine-mediated control of soluble nitrogen mobilization during post-clipping re-growth of white clover (Trifolium repens L.)

A putative contribution of polyamines to the control of peptidase activity expression during re-growth was studied in source organs (roots and stolons) of defoliated white clover (Trifolium repens L.). Endopeptidase activity increased in roots during the first 6 days following complete defoliation, while exopeptidase expression seemed to be restricted to the early hours of re-growth. These changes correlated with an immediate 80% decline in the content of total free polyamines, mainly represented by the diamine cadaverine. The inhibitory capacities of cadaverine and spermine were tested on enzyme activity in vitro in order to elucidate whether the endogenous polyamine level was associated with the cut-induced endopeptidase expression. Cadaverine seemed to inhibit endopeptidase activity of stolons but not root endopeptidase activity. These data support the view that polyamines may play a role in the regulation of peptidase expression in source organs of white clover during post-clipping re-growth. The existence of different endopeptidase isoforms in roots and stolons is discussed in relation to the molecular mechanisms by which polyamines may regulate their activities.Abbreviations AP aminopeptidase - Cad cadaverine - CP carboxypeptidase - EP endopeptidase - PA(s) polyamine(s) - Spm spermine