Intragenic suppressors of temperature-sensitive rne mutations lead to the dissociation of RNase E activity on mRNA and tRNA substrates in Escherichia coli

RNase E of Escherichia coli is an essential endoribonuclease that is involved in many aspects of RNA metabolism. Point mutations in the S1 RNA-binding domain of RNase E (rne-1 and rne-3071) lead to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay and tRNA maturation. However, it is not clear whether RNase E acts similarly on all kinds of RNA substrates. Here we report the isolation and characterization of three independent intragenic second-site suppressors of the rne-1 and rne-3071 alleles that demonstrate for the first time the dissociation of the in vivo activity of RNase E on mRNA versus tRNA and rRNA substrates. Specifically, tRNA maturation and 9S rRNA processing were restored to wild-type levels in each of the three suppressor mutants (rne-1/172, rne-1/186 and rne-1/187), while mRNA decay and autoregulation of RNase E protein levels remained as defective as in the rne-1 single mutant. Each single amino acid substitution (Gly→Ala at amino acid 172; Phe → Cys at amino acid 186 and Arg → Leu at amino acid 187) mapped within the 5′ sensor region of the RNase E protein. Molecular models of RNase E suggest how suppression may occur

Understanding Policy and Programming on Sex-Selection in Tamil Nadu: Ethnographic and Sociological Reflections

The family-planning programme of Tamil Nadu, largely a female sterilisation campaign, has been applauded as one of the successful public health interventions in India, which had arguably led to the drastic fertility decline in the state. To the state's dismay, however, the fertility decline in Tamil Nadu was also attended by the increasing reports of female infanticide and sex-selective abortion. In its subsequent response, the state in Tamil Nadu introduced specific policy and interventionary measures to curb the practice. In this paper, I critically examine these responses in their local ethnographic contexts to highlight the manner in which family-planning goals get intertwined with the political intervention on the issue of sex-selection. This leads to women's diminishing access to unmet needs for family planning and reproductive health services thereby contributing to further marginalisation of Tamil women

DPEP1 Inhibits Tumor Cell Invasiveness, Enhances Chemosensitivity and Predicts Clinical Outcome in Pancreatic Ductal Adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers worldwide. To identify biologically relevant genes with prognostic and therapeutic significance in PDAC, we first performed the microarray gene-expression profiling in 45 matching pairs of tumor and adjacent non-tumor tissues from resected PDAC cases. We identified 36 genes that were associated with patient outcome and also differentially expressed in tumors as compared with adjacent non-tumor tissues in microarray analysis. Further evaluation in an independent validation cohort (N = 27) confirmed that DPEP1 (dipeptidase 1) expression was decreased (T: N ratio ∼0.1, P<0.01) in tumors as compared with non-tumor tissues. DPEP1 gene expression was negatively correlated with histological grade (Spearman correlation coefficient = −0.35, P = 0.004). Lower expression of DPEP1 in tumors was associated with poor survival (Kaplan Meier log rank) in both test cohort (P = 0.035) and validation cohort (P = 0.016). DPEP1 expression was independently associated with cancer-specific mortality when adjusted for tumor stage and resection margin status in both univariate (hazard ratio = 0.43, 95%CI = 0.24–0.76, P = 0.004) and multivariate analyses (hazard ratio = 0.51, 95%CI = 0.27–0.94, P = 0.032). We further demonstrated that overexpression of DPEP1 suppressed tumor cells invasiveness and increased sensitivity to chemotherapeutic agent Gemcitabine. Our data also showed that growth factor EGF treatment decreased DPEP1 expression and MEK1/2 inhibitor AZD6244 increased DPEP1 expression in vitro, indicating a potential mechanism for DPEP1 gene regulation. Therefore, we provide evidence that DPEP1 plays a role in pancreatic cancer aggressiveness and predicts outcome in patients with resected PDAC. In view of these findings, we propose that DPEP1 may be a candidate target in PDAC for designing improved treatments

Human antimicrobial protein hCAP18/LL-37 promotes a metastatic phenotype in breast cancer

International audienc

Fluorescence in situ hybridisation analysis of chromosomal aberrations in gastric tissue: the potential involvement of Helicobacter pylori

In this series of experiments, a novel protocol was developed whereby gastric cells were collected using endoscopic cytology brush techniques, and prepared, such that interphase fluorescence in situ hybridization (FISH) could be performed. In total, 80 distinct histological samples from 37 patients were studied using four chromosome probes (over 32 000 cells analysed). Studies have previously identified abnormalities of these four chromosomes in upper GI tumours. Using premalignant tissues, we aimed to determine how early in Correa's pathway to gastric cancer these chromosome abnormalities occurred. Aneuploidy of chromosomes 4, 8, 20 and 17(p53) was detected in histologically normal gastric mucosa, as well as in gastritis, intestinal metaplasia, dysplasia and cancer samples. The levels of aneuploidy increased as disease severity increased. Amplification of chromosome 4 and chromosome 20, and deletion of chromosome 17(p53) were the more common findings. Hence, a role for these abnormalities may exist in the initiation of, and the progression to, gastric cancer. Helicobactor pylori infection was determined in premalignant tissue using histological analysis and PCR technology. Detection rates were comparable. PCR was used to subtype H. pylori for CagA status. The amplification of chromosome 4 in gastric tissue was significantly more prevalent in H. pylori-positive patients (n=7) compared to H. pylori-negative patients (n=11), possibly reflecting a role for chromosome 4 amplification in H. pylori-induced gastric cancer. The more virulent CagA strain of H. pylori was associated with increased disease pathology and chromosomal abnormalities, although numbers were small (CagA+ n=3, CagA− n=4). Finally, in vitro work demonstrated that the aneuploidy induced in a human cell line after exposure to the reactive oxygen species (ROS) hydrogen peroxide was similar to that already shown in the gastric cancer pathway, and may further strengthen the hypothesis that H. pylori causes gastric cancer progression via an ROS-mediated mechanism

Picard physicians' perspectives about the Legislative Decree on the physical activity prescription

International audienceObjectives. - The present study focused on the impact of the French legislative decree on the physical activity (PA) prescription on the promoting PA during the doctor's consultation and the perceived criteria such limiting factors of the PA prescription for a liberal practitioners' population (MD). Methods. - This exploratory qualitative study involved semi-structured interviews in a sample of MD practiced their liberal activity in the Picard territory of the region Hauts-de-France. Results. - Interviews were carried out with 14 voluntary MD. All MD interviewed were limited to giving oral PA advice as secondary and as tertiary preventions. Therefore, the PA promotion remained often superficial and rushed. Although they believed in the health benefits of regular PA, all practitioners found that the medical consultation is too short and their scientific knowledge on the PA was insufficient to correctly promote an adapted PA. No consensus was found among respondents as to the legislative decree effect on promoting PA due to the gap of information and support assistance. Conclusions. - There seems to be no generally agreed upon the French legislative decree on the physical activity prescription. Perceived major barriers relating to the regular PA promotion, supplied on prescription or under oral advice, showed the weakness of promoting health policy by regular physical activity. (C) 2021 Elsevier Masson SAS. All rights reserved

Variance Reduction Techniques for Estimating Value-at-Risk

This paper describes, analyzes and evaluates an algorithm for estimating portfolio loss probabilities using Monte Carlo simulation.Obtaining accurate estimates of such loss probabilities is essential to calculating value-at-risk, which is a quantile of the loss distribution. The method employs a quadratic ("delta-gamma") approximation to the change in portfolio value to guide the selection of effective variance reduction techniques;specifically importance sampling and stratified sampling.If the approximation is exact, then the importance sampling is shown to be asymptotically optimal.Numerical results indicate that an appropriate combination of importance sampling and stratified sampling can result in large variance reductions when estimating the probability of large portfolio losses.value-at-risk, monte carlo, simulation, variance reduction technique, importance sampling, stratified sampling, rare event