Tetrahydropyrazolo[1,5-a]Pyrimidine-3-Carboxamide and N-Benzyl-6′,7′-Dihydrospiro[Piperidine-4,4′-Thieno[3,2-c]Pyran] analogues with bactericidal efficacy against Mycobacterium tuberculosis targeting MmpL3

Mycobacterium tuberculosis is a major human pathogen and the causative agent for the pulmonary disease, tuberculosis (TB). Current treatment programs to combat TB are under threat due to the emergence of multi-drug and extensively-drug resistant TB. As part of our efforts towards the discovery of new anti-tubercular leads, a number of potent tetrahydropyrazolo[1,5-a]pyrimidine-3-ca​rboxamide(THPP) and N-benzyl-6′,7′-dihydrospiro[piperidine-4,​4′-thieno[3,2-c]pyran](Spiro) analogues were recently identified against Mycobacterium tuberculosis and Mycobacterium bovis BCG through a high-throughput whole-cell screening campaign. Herein, we describe the attractive in vitro and in vivo anti-tubercular profiles of both lead series. The generation of M. tuberculosis spontaneous mutants and subsequent whole genome sequencing of several resistant mutants identified single mutations in the essential mmpL3 gene. This ‘genetic phenotype’ was further confirmed by a ‘chemical phenotype’, whereby M. bovis BCG treated with both the THPP and Spiro series resulted in the accumulation of trehalose monomycolate. In vivo efficacy evaluation of two optimized THPP and Spiro leads showed how the compounds were able to reduce >2 logs bacterial cfu counts in the lungs of infected mice

Mycobacterium tuberculosis Responds to Chloride and pH as Synergistic Cues to the Immune Status of its Host Cell

PubMed ID: 23592993This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Synthesis, characterization, biological and electrical conductivity studies of some Schiff base metal complexes

Metal complexes of VO(IV), Zr(IV), Th(IV) and UO2(VI) with Schiff base ligands derived from 4-nitrobenzoylhydrazide with 2-hydroxy-5-methylacetophenone (H2L1) or 2-hydroxy-5-chloroacetophenone (H2L2) have been prepared. All the complexes have been characterized on the basis of elemental analyses, magnetic susceptibility measurement, electronic and IR spectra and thermogravimetric analysis. The IR spectral data suggested that the ligands behave as dibasic tridentate moiety towards the central metal ion coordinating through phenolic oxygen, enolic oxygen and azomethine nitrogen atoms. The elemental analyses show a 1:1 metal:ligand stoichiometry for all the complexes except Th(IV) which has 1:2 stoichiometry. The thermal analysis evidenced that thermal transformations of complexes are processes according to TG curves including dehydration, thermolysis and oxidative degradation of Schiff base. The final product of decomposition is the most stable metallic oxide. The kinetic analysis of the thermogravimetric data was performed by using the Coats-Redfern method. Solid state electrical conductivity of the complexes has been measured in their compressed pellet form over a 310-413 K temperature range. All the complexes show semiconducting behavior as their conductivity increases with increasing temperature and a function of ionic size. All the complexes along with ligands were also screened for their antibacterial and antifungal activities. KEY WORDS: Aroylhydrazones, Metal complexes, Biological activity, TGA, Electrical conductivity Bull. Chem. Soc. Ethiop. 2014, 28(2), 255-264.DOI: http://dx.doi.org/10.4314/bcse.v28i2.

Rac1 GTPase and the Rac1 exchange factor Tiam1 associate with Wnt-responsive promoters to enhance beta-catenin/TCF-dependent transcription in colorectal cancer cells

<p>Abstract</p> <p>Background</p> <p>β-catenin is a key mediator of the canonical Wnt pathway as it associates with members of the T-cell factor (TCF) family at Wnt-responsive promoters to drive the transcription of Wnt target genes. Recently, we showed that Rac1 GTPase synergizes with β-catenin to increase the activity of a TCF-responsive reporter. This synergy was dependent on the nuclear presence of Rac1, since inhibition of its nuclear localization effectively abolished the stimulatory effect of Rac1 on TCF-responsive reporter activity. We hypothesised that Rac1 plays a direct role in enhancing the transcription of endogenous Wnt target genes by modulating the β-catenin/TCF transcription factor complex.</p> <p>Results</p> <p>We employed chromatin immunoprecipitation studies to demonstrate that Rac1 associates with the β-catenin/TCF complex at Wnt-responsive promoters of target genes. This association served to facilitate transcription, since overexpression of active Rac1 augmented Wnt target gene activation, whereas depletion of endogenous Rac1 by RNA interference abrogated this effect. In addition, the Rac1-specific exchange factor, Tiam1, potentiated the stimulatory effects of Rac1 on the canonical Wnt pathway. Tiam1 promoted the formation of a complex containing Rac1 and β-catenin. Furthermore, endogenous Tiam1 associated with endogenous β-catenin, and this interaction was enhanced in response to Wnt3a stimulation. Intriguingly, Tiam1 was recruited to Wnt-responsive promoters upon Wnt3a stimulation, whereas Rac1 was tethered to TCF binding elements in a Wnt-independent manner.</p> <p>Conclusion</p> <p>Taken together, our results suggest that Rac1 and the Rac1-specific activator Tiam1 are components of transcriptionally active β-catenin/TCF complexes at Wnt-responsive promoters, and the presence of Rac1 and Tiam1 within these complexes serves to enhance target gene transcription. Our results demonstrate a novel functional mechanism underlying the cross-talk between Rac1 and the canonical Wnt signalling pathway.</p

Exploring the Mode of Action of Bioactive Compounds by Microfluidic Transcriptional Profiling in Mycobacteria

10.1371/journal.pone.0069191PLoS ONE87-POLN

Elucidation of the Mode of Action of a New Antibacterial Compound Active against Staphylococcus aureus and Pseudomonas aeruginosa.

Nosocomial and community-acquired infections caused by multidrug resistant bacteria represent a major human health problem. Thus, there is an urgent need for the development of antibiotics with new modes of action. In this study, we investigated the antibacterial characteristics and mode of action of a new antimicrobial compound, SPI031 (N-alkylated 3, 6-dihalogenocarbazol 1-(sec-butylamino)-3-(3,6-dichloro-9H-carbazol-9-yl)propan-2-ol), which was previously identified in our group. This compound exhibits broad-spectrum antibacterial activity, including activity against the human pathogens Staphylococcus aureus and Pseudomonas aeruginosa. We found that SPI031 has rapid bactericidal activity (7-log reduction within 30 min at 4x MIC) and that the frequency of resistance development against SPI031 is low. To elucidate the mode of action of SPI031, we performed a macromolecular synthesis assay, which showed that SPI031 causes non-specific inhibition of macromolecular biosynthesis pathways. Liposome leakage and membrane permeability studies revealed that SPI031 rapidly exerts membrane damage, which is likely the primary cause of its antibacterial activity. These findings were supported by a mutational analysis of SPI031-resistant mutants, a transcriptome analysis and the identification of transposon mutants with altered sensitivity to the compound. In conclusion, our results show that SPI031 exerts its antimicrobial activity by causing membrane damage, making it an interesting starting point for the development of new antibacterial therapies

Discovery of Q203, a potent clinical candidate for the treatment of tuberculosis

New therapeutic strategies are needed to combat the tuberculosis pandemic and the spread of multidrug-resistant (MDR) and extensively drug-resistant (XDR) forms of the disease, which remain a serious public health challenge worldwide1, 2. The most urgent clinical need is to discover potent agents capable of reducing the duration of MDR and XDR tuberculosis therapy with a success rate comparable to that of current therapies for drug-susceptible tuberculosis. The last decade has seen the discovery of new agent classes for the management of tuberculosis3, 4, 5, several of which are currently in clinical trials6, 7, 8. However, given the high attrition rate of drug candidates during clinical development and the emergence of drug resistance, the discovery of additional clinical candidates is clearly needed. Here, we report on a promising class of imidazopyridine amide (IPA) compounds that block Mycobacterium tuberculosis growth by targeting the respiratory cytochrome bc1 complex. The optimized IPA compound Q203 inhibited the growth of MDR and XDR M. tuberculosis clinical isolates in culture broth medium in the low nanomolar range and was efficacious in a mouse model of tuberculosis at a dose less than 1 mg per kg body weight, which highlights the potency of this compound. In addition, Q203 displays pharmacokinetic and safety profiles compatible with once-daily dosing. Together, our data indicate that Q203 is a promising new clinical candidate for the treatment of tuberculosis

Technical Paper Session I-B - CIGSeS and CIGS2 Thin Film Solar Cells on Flexible Foils for Space Power

The objective of the research is to develop flexible, lightweight, radiationresistant, high-specific-power, highly efficient CuIn1-xGaxSe2-ySy (CIGSeS) and CuIn1- xGaxS2 (CIGS2) thin-film solar cells for space electric power. The near optimum bandgap, potential for higher specific power, and superior radiation resistance make this technology an ideal candidate for space electric power. The superior radiation resistance of CIGSeS thin-film solar cells relative to the conventional silicon and gallium arsenide single-crystal cells in the space radiation environment would extend mission lifetimes substantially. The conventional rigid Si and GaAs cells must be folded in an accordion style for deployment space. This can cause problems of opening up and folding of the solar array as has happened recently with the International Space Station. On the other hand, the flexible solar cells and modules can be packaged and rolled out more easily. The stainless steel and titanium foil substrate materials are capable of withstanding high temperatures required for preparing good quality CIGSeS absorber layer. They also do not sag easily and hence do not require rigidizing as is the case with plastic sheet substrates. The CIGSeS absorber film is prepared by selenization/sulfurization of DC magnetron sputter-deposited CuGa, In metallic precursors on 10 cm x 10 cm metallic foil substrate coated with molybdenum back contact layer. CdS heterojunction partner is deposited by chemical bath deposition. Transparent and conducting bilayer of intrinsic ZnO and aluminum doped ZnO:Al is deposited by RF magnetron sputtering. Cells are completed by depositing Ni/Al front contact fingers by thermal evaporation. The sputtering technique utilized in the preparation of solar cells provides an added advantage of facilitating easy scale-up of the laboratory size cells for economic large-area manufacture by the roll-to-roll process. Chemical composition, crystallographic structure and morphology of CIGSeS thin films are analyzed by energy dispersive spectroscopy, Auger electron spectroscopy, X-ray diffraction, scanning electron microscopy and transmission electron microscopy. The photovoltaic properties of completed cells are studied by measurement of current-voltage characteristics and quantum efficiency. Best efficiencies of 10.4% under AM 1.5 conditions and 8.84% under AM 0 conditions were achieved on small-area CIGS2 thin-film solar cells

Association of apolipoprotein E polymorphisms and dietary factors in colorectal cancer

ApoE single nucleotide polymorphisms (SNPs) Cys112Arg (Epsilon-4), and Arg158Cys (Epsilon-2) have been implicated in cardiovascular and Alzheimer's disease, but their role in colorectal cancer (CRC) has not been extensively studied. We investigated whether ApoE polymorphisms alone or in combination with dietary factors selectively contribute to mismatch-repair (MMR) proficient (microsatellite stable/low or MSS/L) vs deficient (microsatellite unstable or MSI-H) CRCs. We carried out a case–control study with 906 CRC cases and 911 unaffected controls to examine the associations between ApoE polymorphisms and dietary factors and assessed their contribution to MSS/L and MSI-H CRCs. We used unconditional logistic regression to evaluate the associations between ApoE SNPs, tumour MSI status, and dietary factors after adjusting for age and sex. All statistical tests were two-sided. No significant differences in ApoE genotype frequencies were observed between CRC cases and unaffected controls. We observed that increased dietary intake of total fat, saturated fat, cholesterol, and red meat was significantly associated with CRC. Among non-ApoE4 carriers, 2–4 and >4 red meat servings/week were associated with developing MSS/L CRC (OR=1.51, 95% CI 1.10–2.07 and OR=1.80, 95% CI 1.30–2.48, respectively), whereas among ApoE4 allele carriers, four or more red meat servings/week were associated with MSI-H CRC (OR=4.62, 95% CI 1.20–17.77) when compared with the controls. ApoE isoforms modulate the risk of MSI-H and MSS/L CRCs among high red meat consumers

Edaxadiene: A New Bioactive Diterpene from Mycobacterium tuberculosis

Mycobacterium tuberculosis remains a widespread and devastating human pathogen. Presented here is the characterization of an atypical class I diterpene cyclase from M. tuberculosis that catalyzes an unusual cyclization reaction in converting the known M. tuberculosis metabolite halimadienyl diphosphate to a further cyclized novel diterpene, which we have termed edaxadiene, as it directly inhibits maturation of the phagosomal compartment in which the bacterium is taken up during infection