Additional Routes to Staphylococcus aureus Daptomycin Resistance as Revealed by Comparative Genome Sequencing, Transcriptional Profiling, and Phenotypic Studies

Daptomycin is an extensively used anti-staphylococcal agent due to the rise in methicillin-resistant Staphylococcus aureus, but the mechanism(s) of resistance is poorly understood. Comparative genome sequencing, transcriptomics, ultrastructure, and cell envelope studies were carried out on two relatively higher level (4 and 8 mu g/ml(-1)) laboratory-derived daptomycin-resistant strains (strains CB1541 and CB1540 respectively) compared to their parent strain (CB1118; MW2). Several mutations were found in the strains. Both strains had the same mutations in the two-component system genes waIK and agrA. In strain CB1540 mutations were also detected in the ribose phosphate pyrophosphokinase (prs) and polyribonucleotide nucleotidyltransferase genes (pnpA), a hypothetical protein gene, and in an intergenic region. In strain CB1541 there were mutations in clpP, an ATP-dependent protease, and two different hypothetical protein genes. The strain CB1540 transcriptome was characterized by upregulation of cap (capsule) operon genes, genes involved in the accumulation of the compatible solute glycine betaine, ure genes of the urease operon, and mscL encoding a mechanosensitive chanel. Downregulated genes included smpB, femAB and femH involved in the formation of the pentaglycine interpeptide bridge, genes involved in protein synthesis and fermentation, and spa encoding protein A. Genes altered in their expression common to both transcriptomes included some involved in glycine betaine accumulation, mscL, ure genes, femH, spa and smpB. However, the CB1541 transcriptome was further characterized by upregulation of various heat shock chaperone and protease genes, consistent with a mutation in clpP, and lytM and sceD. Both strains showed slow growth, and strongly decreased autolytic activity that appeared to be mainly due to decreased autolysin production. In contrast to previous common findings, we did not find any mutations in phospholipid biosynthesis genes, and it appears there are multiple pathways to and factors in daptomycin resistance

An integrated, bottom-up approach to evaluate the role of bioenergy with carbon capture and storage (BECCS) in achieving net-zero energy systems

Early-stage deployment of BECCS, supported by integrated policies and financing mechanisms to suit the socioeconomic and environmental context, could help achieve net-zero energy systems at small scales and further evaluate their feasibility in the long run

Strategies for coping with the costs of inpatient care: a mixed methods study of urban and rural poor in Vadodara District, Gujarat, India

Background In India, coping mechanisms for inpatient care costs have been explored in rural areas, but seldom among urbanites. This study aims to explore and compare mechanisms employed by the urban and rural poor for coping with inpatient expenditures, in order to help identify formal mechanisms and policies to provide improved social protection for health care

Factors Affecting Male Pink Bollworm Moth Catches in Gossyplure Baited Traps

In an experiment conducted in cotton fields at Hissar, Haryana (India) during the year 1978–79, it was observed that the most effective trap for the capture of male pink bolworm moths using sex pheromone "gossyplure" was metal trap in which either rubber stopper or plastic tube was used as pheromone dispenser and either liquid paraffiIl or cottOl1 seed oil was used as moth sticking material. Maximum moths were captured when the traps were kept at the plant canopy level and at a distance of 60 or 40 metres

Role of acute viral hepatitis as a confounding factor in antituberculosis treatment induced hepatotoxicity

Background & Objective: Drug induced hepatotoxicity (DIH) is an important and commonly encountered adverse effect with antituberculosis (anti-TB) treatment. Acute viral hepatitis (AVH) is an important confounding reason which clinically, biochemically and histologically mimics DIH. Methods: The contributory role of acute viral hepatitis as a confounding factor in patients with normal baseline liver functions who developed acute hepatitis while receiving short-course anti-TB treatment was prospectively studied. The sera of all patients who developed acute hepatitis were analysed for markers for hepatitis A, B, C and E viruses. Results: Viral hepatitis was present in 15 of the 102 (14.7%) patients who developed acute hepatitis while receiving anti-TB treatment with hepatitis E virus being the most common cause Later onset of acute hepatitis [58 (5-133) vs. 26 (3-221) days; P=0.04], large elevations in aspartate aminotransferase (AST) [371 (30-2643) vs. 212 (63-1990 IU/l); P=0.03] and alanine aminotransferase (ALT) [388 (31-2997) vs. 225 (52- 1670 IU/l); P= 0.002] and a longer time for normalization of deranged liver functions [36.7 ± 13.3 vs. 24.5 ± 19.3 days; P=0.02] indicated acute viral hepatitis as the cause of liver function derangement. Interpretation & Conclusion: Our findings showed AVH in 14.7 per cent patients who developed hepatotoxicity while an anti-TB treatment. Therefore, in endemic areas, viral hepatitis should be sought after and excluded in all patients suspected to have DIH before attributing the hepatotoxic effect to the anti-TB drugs

miR-10a is aberrantly overexpressed in Nucleophosmin1 mutated acute myeloid leukaemia and its suppression induces cell death

<p>Abstract</p> <p>Background</p> <p>Acute myeloid leukaemia (AML) with nucleophosmin-1 (<it>NPM1</it>) mutation is a major subtype of AML. The <it>NPM1 </it>mutation induces a myeloproliferative disorder, but evidence indicates that other insults are necessary for the development of AML. We utilised microRNA microarrays and functional assays to determine if microRNA dysregulation could be involved in the pathogenesis of in <it>NPM1 </it>mutated (<it>NPM1^mut</it>)-AML.</p> <p>Results</p> <p>We used a stringent locked nucleic acid (LNA) based microRNA microarray platform to profile bone marrow samples of patients with normal karyotype AML. A panel of five microRNAs dichotomised AML patients according to their <it>NPM1 </it>mutational status. miR-10a, let-7b and let-7c were significantly over-expressed, while miR-130a and miR-335 were under-expressed in <it>NPM1^mut</it>-AML when compared to <it>NPM1^wildtype</it>-AML. Of these, miR-10a is the most differentially expressed in <it>NPM1^mut</it>-AML versus <it>NPM1^wildtype</it>-AML (> 10 fold higher as confirmed by qRT-PCR). To investigate the functions of miR-10a, the OCI-AML3 cell line was utilised, which is the only commercially available cell line bearing <it>NPM1^mut</it>. OCI-AML3 cells were firstly demonstrated to have a similarly high miR-10a expression to primary <it>NPM1^mut</it>-AML patient samples. Inhibition of miR-10a expression by miRCURY LNA Inhibitors (Exiqon) in these cells resulted in increased cell death as assessed by MTS, cell cycle and Annexin-V assays and reduced clonogenic capacity, indicative of an involvement in leukaemic cell survival. <it>In silico </it>filtering of bioinformatically predicted targets of miR-10a identified a number of potential mRNA targets with annotated functions in haematopoiesis, cell growth and apoptosis. Lucferase reporter assays confirmed a number of these putative tumorogenic genes that are miR-10a suppressible including <it>KLF4 </it>and <it>RB1CC1</it>. This provides a potential mechanism for the pathogenic role of miR-10a in <it>NPM1^mut</it>-AML.</p> <p>Conclusions</p> <p>This study provides, for the first time, <it>in vitro </it>evidence of a pro-survival role of miR-10a in <it>NPM1^mut</it>-AML, that it may contribute to the pathogenesis of <it>NPM1^mut</it>-AML and identifies putative tumorogenic targets.</p

Diagnostic accuracy of non-invasive tests for advanced fibrosis in patients with NAFLD: an individual patient data meta-analysis

Objective Liver biopsy is still needed for fibrosis staging in many patients with non-alcoholic fatty liver disease. The aims of this study were to evaluate the individual diagnostic performance of liver stiffness measurement by vibration controlled transient elastography (LSM-VCTE), Fibrosis-4 Index (FIB-4) and NAFLD (non-alcoholic fatty liver disease) Fibrosis Score (NFS) and to derive diagnostic strategies that could reduce the need for liver biopsies. Design Individual patient data meta-analysis of studies evaluating LSM-VCTE against liver histology was conducted. FIB-4 and NFS were computed where possible. Sensitivity, specificity and area under the receiver operating curve (AUROC) were calculated. Biomarkers were assessed individually and in sequential combinations. Results Data were included from 37 primary studies (n=5735; 45% women; median age: 54 years; median body mass index: 30 kg/m2; 33% had type 2 diabetes; 30% had advanced fibrosis). AUROCs of individual LSM-VCTE, FIB-4 and NFS for advanced fibrosis were 0.85, 0.76 and 0.73. Sequential combination of FIB-4 cut-offs (<1.3; ≥2.67) followed by LSM-VCTE cut-offs (<8.0; ≥10.0 kPa) to rule-in or rule-out advanced fibrosis had sensitivity and specificity (95% CI) of 66% (63–68) and 86% (84–87) with 33% needing a biopsy to establish a final diagnosis. FIB-4 cut-offs (<1.3; ≥3.48) followed by LSM cut-offs (<8.0; ≥20.0 kPa) to rule out advanced fibrosis or rule in cirrhosis had a sensitivity of 38% (37–39) and specificity of 90% (89–91) with 19% needing biopsy. Conclusion Sequential combinations of markers with a lower cut-off to rule-out advanced fibrosis and a higher cut-off to rule-in cirrhosis can reduce the need for liver biopsies

An Evolutionary Reduction Principle for Mutation Rates at Multiple Loci

A model of mutation rate evolution for multiple loci under arbitrary selection is analyzed. Results are obtained using techniques from Karlin (1982) that overcome the weak selection constraints needed for tractability in prior studies of multilocus event models. A multivariate form of the reduction principle is found: reduction results at individual loci combine topologically to produce a surface of mutation rate alterations that are neutral for a new modifier allele. New mutation rates survive if and only if they fall below this surface - a generalization of the hyperplane found by Zhivotovsky et al. (1994) for a multilocus recombination modifier. Increases in mutation rates at some loci may evolve if compensated for by decreases at other loci. The strength of selection on the modifier scales in proportion to the number of germline cell divisions, and increases with the number of loci affected. Loci that do not make a difference to marginal fitnesses at equilibrium are not subject to the reduction principle, and under fine tuning of mutation rates would be expected to have higher mutation rates than loci in mutation-selection balance. Other results include the nonexistence of 'viability analogous, Hardy-Weinberg' modifier polymorphisms under multiplicative mutation, and the sufficiency of average transmission rates to encapsulate the effect of modifier polymorphisms on the transmission of loci under selection. A conjecture is offered regarding situations, like recombination in the presence of mutation, that exhibit departures from the reduction principle. Constraints for tractability are: tight linkage of all loci, initial fixation at the modifier locus, and mutation distributions comprising transition probabilities of reversible Markov chains.Comment: v3: Final corrections. v2: Revised title, reworked and expanded introductory and discussion sections, added corollaries, new results on modifier polymorphisms, minor corrections. 49 pages, 64 reference

Impact of COVID-19 Pandemic on Vegetable Sector and its Allies

The novel coronavirus 2019 (COVID-19), has created an unanticipated pandemic, that has triggered severe panic among individuals worldwide. In this direction, countries are maximizing their efforts to fight the virus and reduce infection. Vegetables due to the presence of several bioactive compounds could help build and maintain immunity against degenerative diseases and COVID-19. Therefore, sustainable vegetable supply is needed continuously. For this to happen placidly vegetable producer and vegetable sector allies must work in close cooperation with each other. This document deals with the impact of COVID-19 pandemic on vegetable sector and its allies