Benzothiazinones kill Mycobacterium tuberculosis by blocking arabinan synthesis

New drugs are required to counter the tuberculosis (TB) pandemic. Here, we describe the synthesis and characterization of 1,3-benzothiazin-4-ones (BTZs), a new class of antimycobacterial agents that kill Mycobacterium tuberculosis in vitro, ex vivo, and in mouse models of TB. Using genetics and biochemistry, we identified the enzyme decaprenylphosphoryl-beta-d-ribose 2'-epimerase as a major BTZ target. Inhibition of this enzymatic activity abolishes the formation of decaprenylphosphoryl arabinose, a key precursor that is required for the synthesis of the cell-wall arabinans, thus provoking cell lysis and bacterial death. The most advanced compound, BTZ043, is a candidate for inclusion in combination therapies for both drug-sensitive and extensively drug-resistant TB

Numerical simulation of a brick wall containing solar heated water channels in cold regions

Maintenance of indoor comfort temperatures in cold regions especially during winters requires huge energy requirements. However, solar energy can be utilized to reduce this power requirement. The present paper deals with an explicit transient numerical solution of a north facing brick wall in an extremely cold region of Leh (India) containing rectangular sectioned PVC channels along the length of wall filled with water raised to 20 ºC with solar water heater for a typical day in the month of December and another day in the month of May on 24 hours based variable ambient conditions. As compared to a similar brick wall without water channels, average rate of heat transfer from within the simulated indoor environment to the inside surface of wall was calculated to be reduced by 44.6% in December and 29.6% in May so as to maintain a uniform and steady comfort indoor temperature of 26 ºC. 

Marangoni convection in a relatively hotter or cooler liquid layer with free boundaries

In this paper, we study the onset of cellular convection in a horizontal fluid layer heated from below, with a free-slip boundary condition at the bottom when the driving mechanism is surface tension at the upper free surface, in the light of the modified analysis of Banerjee et al. (Jour. Math. & Phys. Sci., 1983, 17, 603). This leads to a formulation of the problem which depends upon whether the liquid layer is relatively hotter or cooler. It is found that the phenomenon of surface tension driven instability problems should not only depend upon the Marangoni number which is proportional to the maintained temperature differences across the layer but also upon another parameter that arises due to variation in the specific heat at constant volume on account of the variations in temperature. Numerical results are obtained for the problem wherein the lower free boundary is perfectly thermally conducting

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Generation of diploid <it>Pichia pastoris</it> strains by mating and their application for recombinant protein production

Abstract Background Yeast mating provides an efficient means for strain and library construction. However, biotechnological applications of mating in the methylotrophic yeast Pichia pastoris have been hampered because of concerns about strain stability of P. pastoris diploids. The aim of the study reported here is to investigate heterologous protein expression in diploid P. pastoris strains and to evaluate diploid strain stability using high cell density fermentation processes. Results By using a monoclonal antibody as a target protein, we demonstrate that recombinant protein production in both wild-type and glycoengineered P. pastoris diploids is stable and efficient during a nutrient rich shake flask cultivation. When diploid strains were cultivated under bioreactor conditions, sporulation was observed. Nevertheless, both wild-type and glycoengineered P. pastoris diploids showed robust productivity and secreted recombinant antibody of high quality. Specifically, the yeast culture maintained a diploid state for 240 h post-induction phase while protein titer and N-linked glycosylation profiles were comparable to that of a haploid strain expressing the same antibody. As an application of mating, we also constructed an antibody display library and used mating to generate novel full-length antibody sequences. Conclusions To the best of our knowledge, this study reports for the first time a comprehensive characterization of recombinant protein expression and fermentation using diploid P. pastoris strains. Data presented here support the use of mating for various applications including strain consolidation, variable-region glycosylation antibody display library, and process optimization.</p

Whole cell screen based identification of spiropiperidines with potent antitubercular properties

Abstract Whole cell based screens to identify hits against Mycobacterium tuberculosis (Mtb), carried out under replicating and non-replicating (NRP) conditions, resulted in the identification of multiple, novel but structurally related spiropiperidines with potent antitubercular properties. These compounds could be further classified into three classes namely 3-(3-aryl-1,2,4-oxadiazol-5-yl)-1′-alkylspiro[indene-1,4′-piperidine] (abbr. spiroindenes), 4-(3-aryl-1,2,4-oxadiazol-5-yl)-1′-alkylspiro[chromene-2,4′-piperidine] (abbr. spirochromenes) and 1′-benzylspiro[indole-1,4′-piperidin]-2(1H)-one (abbr. spiroindolones). Spiroindenes showed ≥4 log10 kill (at 2-12 μM) on replicating Mtb, but were moderately active under non replicating conditions. Whole genome sequencing efforts of spiroindene resistant mutants resulted in the identification of I292L mutation in MmpL3 (Mycobacterial membrane protein Large), required for the assembly of mycolic acid into the cell wall core of Mtb. MIC modulation studies demonstrated that the mutants were cross-resistant to spirochromenes but not to spiroindolones. This Letter describes lead identification efforts to improve potency while reducing the lipophilicity and hERG liabilities of spiroindenes. Additionally, as deduced from the SAR studies, we provide insights regarding the new chemical opportunities that the spiroindolones can offer to the TB drug discovery initiatives