Characterization of late polyoma mRNA

Polyoma-infected mouse kidney cell cultures were labeled with [(3)H]uridine for 3 h late in the lytic cycle (26 to 29 h after infection) and RNA was extracted from cytoplasm and nuclei and from isolated polyribosomes. Sedimentation velocity analysis in sucrose gradients showed that polyoma-specific "giant" and 26S RNAs are present exclusively in the nucleus. RNA associated with cytoplasmic polyribosomes was analyzed by sedimentation in aqueous sucrose density gradients and dimethylsulfoxide sucrose gradients, as well as by polyacrylamide gel electrophoresis. Polyoma-specific RNA in polyribosomes consists of at least two classes, with sedimentation coefficients of 16 (major fraction) and 19S (minor fraction) in aqueous sucrose gradients and 15 and 17S, respectively, in dimethylsulfoxide gradients. Estimates based on dimethylsulfoxide gradient and analysis suggest a molecular weight of approximately 500,000 for 16S RNA and 700,000 for 19S RNA. These polyoma RNAs seem to undergo reversible conformational changes under the different conditions of analysis. We cannot exclude the possibility that they contain more than one molecular species

Tissue-specific and ubiquitous factors binding next to the glucocorticoid receptor modulate transcription from the mouse mammary tumor virus promoter

Steroid hormones complexed with their receptors play an essential role in the regulation of mouse mammary tumor virus (MMTV) transcription. However, the need for additional tissue-specific regulatory factors is suggested by the lack of virus expression in liver, in which glucocorticoid receptors are highly abundant, and by the tissue-specific transcription of reporter genes linked to an MMTV long terminal repeat in transgenic mice. In this study, we characterized two distal-region regulatory elements, DRa and DRc, which, together with the distal glucocorticoid receptor binding site (DRb), increased transcription from the MMTV promoter in permissive cells. This was demonstrated by transfection of these sequences (DRa, DRb, and DRc) in different combinations with the natural MMTV promoter in mouse fibroblasts and mammary epithelial cells, followed by quantitative S1 nuclease mapping of the transcripts. We further showed by DNase I footprinting, methylation interference, and gel retardation assays with various nuclear extracts from permissive or nonpermissive tissues and cell lines that the factors binding to the DRa site are distinct and tissue-specific whereas those binding to DRc are ubiquitous

Approach and avoidance movements are unaffected by cognitive conflict: A comparison of the Simon effect and stimulus-response compatibility

Participants in this study reached from central fixation to a lateral position that either contained or was opposite to the stimulus. Cognitive conflict was induced when the stimulus and response directions did not correspond. In the Simon task, the response direction was cued by the color of the lateral stimulus, and corresponding and noncorresponding trials varied randomly in the same block of trials, resulting in high uncertainty and long reaction times (RTs). In the stimulus-response compatibility (SRC) task, participants reached toward or away from the stimulus in separate blocks of trials, resulting in low uncertainty and short RTs. In the SRC task, cognitive conflict in noncorresponding trials slowed down RTs but hardly affected reach trajectories. In the Simon task, both RTs and reach trajectories were strongly influenced by stimulus-response correspondence. Despite the overall longer RTs in the Simon task, reaches were less direct and deviated toward the stimulus in noncorresponding trials. Thus, cognitive conflict was resolved before movement initiation in the SRC task, whereas it leaked into movement execution in the Simon task. Current theories of the Simon effect, such as the gating of response activation or response code decay, are inconsistent with our results. We propose that the SRC task was decomposed as approaching and avoiding the stimulus, which is sustained by stereotyped visuomotor routines. With complex stimulus-response relationships (Simon task), responses had to be coded as leftward and rightward, with more uncertainty about how to execute the action. This uncertainty permitted cognitive conflict to leak into the movement executio

POH1/Rpn11/PSMD14: a journey from basic research in fission yeast to a prognostic marker and a druggable target in cancer cells.

POH1/Rpn11/PSMD14 is a highly conserved protein in eukaryotes from unicellular organisms to human and has a crucial role in cellular homoeostasis. It is a subunit of the regulatory particle of the proteasome, where it acts as an intrinsic deubiquitinase removing polyubiquitin chains from substrate proteins. This function is not only coupled to the translocation of substrates into the core of the proteasome and their subsequent degradation but also, in some instances, to the stabilisation of ubiquitinated proteins through their deubiquitination. POH1 was initially discovered as a functional homologue of the fission yeast gene pad1 <sup>+</sup> , which confers drug resistance when overexpressed. In translational studies, expression of POH1 has been found to be increased in several tumour types relative to normal adjacent tissue and to correlate with tumour progression, higher tumour grade, decreased sensitivity to cytotoxic drugs and poor prognosis. Proteasome inhibitors targeting the core particle of the proteasome are highly active in the treatment of myeloma, and recently developed POH1 inhibitors, such as capzimin and thiolutin, have shown promising anticancer activity in cell lines of solid tumours and leukaemia. Here we give an overview of POH1 function in the cell, of its potential role in oncogenesis and of recent progress in developing POH1-targeting drugs

Successful Treatment of Acute Prostatitis Caused by Multidrug-Resistant Escherichia coli With Tigecycline Monotherapy

We present a successful treatment, with tigecycline monotherapy, of acute prostatitis caused by multidrug-resistant Escherichia coli harboring an NDM-1 carbapemenase along with a CMY-2 cephalosporinase and a TEM ESBL

Increase in methicillin-susceptible Staphylococcus aureus bloodstream infections in Switzerland: a nationwide surveillance study (2008-2021).

PURPOSE An increasing burden of Staphylococcus aureus bloodstream infections (BSI), despite a decrease in the percentage of methicillin-resistant S. aureus (MRSA), was described recently in other European countries. The main aim of this study was to analyse recent temporal trends of S. aureus, methicillin-susceptible S. aureus (MSSA) and MRSA BSI for Switzerland as well as the different linguistic regions within Switzerland. An additional aim was to estimate potential differences among patient-based and epidemiological risk factors. METHODS A retrospective observational study was conducted in Switzerland over a period of 14 years (2008-2021). Trends in S. aureus, MSSA and MRSA BSI were analysed by applying linear regression models. RESULTS Staphylococcus aureus BSI increased by + 30% from 19.7 to 25.6 cases per 100,000 inhabitants between 2008 and 2021 (P < 0.01) in Switzerland. Thereof, MSSA increased by + 37% from 17.8 to 24.4 cases per 100,000 inhabitants (P < 0.01). MRSA decreased from 1.9 to 1.2 cases per 100,000 inhabitants (P < 0.01), which was driven by decreasing incidence in the French-speaking region. MSSA BSI increased significantly (P < 0.01) in both linguistic regions. A further stratification revealed that incidence increased the most in male patients of the age group ≥ 80 years of the German-speaking region. CONCLUSION The increasing health burden of MSSA BSI in Switzerland indicates that not only proportions of resistant microorganisms but also total BSI incidences should be monitored. In addition, data stratification revealed that the increase was mainly driven by an increasing incidence in elderly males of the German-speaking region

Gene transfer into stimulated and unstimulated T lymphocytes by HIV-1-derived lentiviral vectors

Genetic modification of T lymphocytes holds great potential for treatments of cancer, T cell disorders and AIDS. While in the past recombinant murine retroviruses were the vectors of choice for gene delivery to T cells, vectors based on lentiviruses can provide additional benefits. Here, we show that VSV-G pseudotyped HIV 1 vector particles delivering the enhanced green fluorescent protein (EGFP) efficiently transduce human T lymphocytes. Transduction efficiency was optimal when infection included centrifugation of cells with concentrated vector supernatant in the presence of Polybrene. In contrast to previous reports describing murine retrovirus-mediated gene transfer to T lymphocytes, fibronectin did not improve the transduction efficiency of the VSVG-pseudotyped HIV-1 particles. Similar gene transfer efficiencies were observed following stimulation of cells with PHA/IL-2 or anti-CD3i/CD28i antibodies, although greater transgene expression was observed in the latter case. Interestingly, production of vectors in the absence of the accessory proteins Vif, Vpr, Vpu and Nef was accompanied by a 50% decrease in transduction efficiency in activated T cells. Transduction of T cells that were not stimulated before infection was achieved. No transduction of non-prestimulated cells was observed with a GAL V-pseudotyped murine retroviral vector. The requirement for accessory proteins in non-prestimulated cells was more pronounced. Our results have implications for lentiviral vector targeting of other cells of the hematopoietic system including stem cells

Isopropanol at 60% and at 70% are effective against 'isopropanol-tolerant' Enterococcus faecium.

The bactericidal activity of isopropanol was determined against Enterococcus faecium ATCC 6057, ST 796 (isopropanol-tolerant strain) and Enterococcus hirae ATCC 10541 (EN 13727). Isopropanol at 60% and 70% were effective (≥5.38 log10-reduction) in 15 s against all strains but 23% isopropanol was not (<0.99 log10-reduction in ≤15 min). Isopropanol at 70% was tested against E. faecium in the four-field test. Eight millilitres was not effective enough in 1 min (<5 log10-reduction), whilst 16 mL was effective (≥5.85 log10-reduction). Healthcare workers can be reassured that 60% and 70% isopropanol with an appropriate volume are effective against E. faecium

Femmes immigrantes et intersectionnalité: enjeux méthodologiques pour la recherche en santé au Canada

L’article qui suit présente les principaux questionnements méthodologiques relatifs à l’opérationnalisation de l’approche intersectionnelle dans les recherches en santé des femmes immigrantes au Canada. L’identification et la compréhension de l’interaction des différents systèmes d’oppression, la catégorisation de ces intersections et le développement d’une grille d’analyse efficace demeurent les principaux obstacles. Quelques pistes de solution sont par la suite introduites

Distribution of pathogens and antimicrobial resistance in ICU-bloodstream infections during hospitalization: a nationwide surveillance study.

Changing microorganism distributions and decreasing antibiotic susceptibility over the duration of hospitalization have been described for the colonization or infection of selected organ systems. Few data are available on bacteremias in the intensive care unit (ICU) setting. We conducted a nationwide study on bloodstream infection (BSI) using data from the Swiss Centre for Antibiotic Resistance (ANRESIS). We analyzed data on BSI detected in the ICU from hospitals that sent information on a regular basis during the entire study period (2008-2017). We described specific trends of pathogen distribution and resistance during hospitalization duration. We included 6505 ICU- BSI isolates from 35 Swiss hospitals. We observed 2587 possible skin contaminants, 3788 bacteremias and 130 fungemias. The most common microorganism was Escherichia coli (23.2%, 910), followed by Staphylococcus aureus (18.7%, 734) and enterococci (13.1%, 515). Enterococcus spp (p < 0.0001) and Candida spp (p < 0.0001) increased in proportion, whereas E. coli (p < 0.0001) and S. aureus (p < 0.0001) proportions decreased during hospitalization. Resistances against first- and second-line antibiotics increased linearly during hospitalization. Pathogen distribution and antimicrobial resistance in ICU-BSI depends on the duration of the hospitalization. The proportion of enterococcal BSI, candidemia and resistant microorganisms against first- and second-line antibiotics increased during hospitalization