Bid participates in genotoxic drug-induced apoptosis of HeLa cells and is essential for death receptor ligands' apoptotic and synergistic effects

Background: The BH3-only protein Bid is an important component of death receptor-mediated caspase activation. Bid is cleaved by caspase-8 or -10 into t-Bid, which translocates to mitochondria and triggers the release of caspase-activating factors. Bid has also been reported to be cleaved by other proteases. Methodology/Principal Findings: To test the hypothesis that Bid is a central mediator of stress-induced apoptosis, we investigated the effects of a small molecule Bid inhibitor on stress-induced apoptosis, and generated HeLa cells deficient for Bid. Stable knockdown of bid lead to a pronounced resistance to Fas/CD95- and TRAIL-induced caspase activation and apoptosis, and significantly increased clonogenic survival. While Bid-deficient cells were equally sensitive to ER stress-induced apoptosis, they showed moderate, but significantly reduced levels of apoptosis, as well as increased clonogenic survival in response to the genotoxic drugs Etoposide, Oxaliplatin, and Doxorubicin. Similar effects were observed using the Bid inhibitor BI6C9. Interestingly, Bid-deficient cells were dramatically protected from apoptosis when subtoxic concentrations of ER stressors, Etoposide or Oxaliplatin were combined with subtoxic TRAIL concentrations. Conclusions/Significance: Our data demonstrate that Bid is central for death receptor-induced cell death and participates in anti-cancer drug-induced apoptosis in human cervical cancer HeLa cells. They also show that the synergistic effects of TRAIL in combination with either ER stressors or genotoxic anti-cancer drugs are nearly exclusively mediated via an increased activation of Bid-induced apoptosis signalling

Amplified warming of seasonal cold extremes relative to the mean in the Northern Hemisphere extratropics

Cold extremes are anticipated to warm at a faster rate than both hot extremes and average temperatures for much of the Northern Hemisphere. Anomalously warm cold extremes can affect numerous sectors, including human health, tourism and various ecosystems that are sensitive to cold temperatures. Using a selection of global climate models, this paper explores the accelerated warming of seasonal cold extremes relative to seasonal mean temperatures in the Northern Hemisphere extratropics. The potential driving physical mechanisms are investigated by assessing conditions on or prior to the day when the cold extreme occurs to understand how the different environmental fields are related. During winter, North America, Europe and much of Eurasia show amplified warming of cold extremes projected for the late 21st century, compared to the mid-20th century. This is shown to be largely driven by reductions in cold air temperature advection, suggested as a likely consequence of Arctic amplification. In spring and autumn, cold extremes are expected to warm faster than average temperatures for most of the Northern Hemisphere mid-latitudes to high latitudes, particularly Alaska, northern Canada and northern Eurasia. In the shoulder seasons, projected decreases in snow cover and associated reductions in surface albedo are suggested as the largest contributor affecting the accelerated rates of warming in cold extremes. The key findings of this study improve our understanding of the environmental conditions that contribute to the accelerated warming of cold extremes relative to mean temperatures.This study was supported by the Australian Research Council (ARC) Centre of Excellence for Climate Extremes (grant CE170100023). Markus G. Donat received funding from the ARC (grant DE150100456) and the Spanish Ministry for the Economy, Industry and Competitiveness Ramón y Cajal 2017 grant reference RYC-2017-22964. We acknowledge the World Climate Research Programme's Working Group on Coupled Modelling, which is responsible for CMIP, and we thank the climate modelling groups (listed in Table 1 of this paper) for producing and making their model output available.Peer ReviewedPostprint (published version

<研究ノート>西成特区構想の展開と課題 : あいりん地域の新たなセーフティネットづくりを中心に

In this report, Mo(VI) ions are transported from an aqueous donor phase into an aqueous acceptor phase by a newly designed method called as multi dropped liquid membrane (MDLM) system prepared by dissolving TNOA as carrier in kerosene. During the extraction of Mo(VI) ions by the liquid membrane system; 100ppm Mo(VI) solutions as donor phase, buffer solution(pH:9.5) and Na2CO3 in different concentrations as acceptor phase and TNOA diluted by kerosen as organic phase are used.In our experimental work, the effect of temperature by using buffer solution and Na2CO3 in the acceptor phase and effect of concentration of acceptor phase on the extraction of Mo(VI) ions were investigated. Appropriate conditions for Mo(VI) transportation were as follows: pH of donor phase is 2.00, concentration of TNOA is 0.005M, 1.00M Na2CO3 as acceptor phase, and flux rate is 50mL/min. Besides, Mo(VI) ion transportation is consecutive first order irreversible reaction and the transportation of Mo(VI) ions is diffusion controlled process. The kinetic parameters (k1, k2, Rm(max), tmax, Jd(max), Ja(max)) were calculated for the interface reactions assuming two consecutive, irreversible first-order reactions

Relativistic simulations of rotational core collapse. I. Methods, initial models, and code tests

We describe an axisymmetric general relativistic code for rotational core collapse. The code evolves the coupled system of metric and fluid equations using the ADM 3+1 formalism and a conformally flat metric approximation of the Einstein equations. The relativistic hydrodynamics equations are formulated as a first-order flux-conservative hyperbolic system and are integrated using high-resolution shock-capturing schemes based on Riemann solvers. We assess the quality of the conformally flat metric approximation for relativistic core collapse and present a comprehensive set of tests which the code successfully passed. The tests include relativistic shock tubes, the preservation of the rotation profile and of the equilibrium of rapidly and differentially rotating neutron stars (approximated as rotating polytropes), spherical relativistic core collapse, and the conservation of rest-mass and angular momentum in dynamic spacetimes. The application of the code to relativistic rotational core collapse, with emphasis on the gravitational waveform signature, is presented in an accompanying paper.Comment: 18 pages, 12 figure

Bid Participates in Genotoxic Drug-Induced Apoptosis of HeLa Cells and Is Essential for Death Receptor Ligands' Apoptotic and Synergistic Effects

Background: The BH3-only protein Bid is an important component of death receptor-mediated caspase activation. Bid is cleaved by caspase-8 or -10 into t-Bid, which translocates to mitochondria and triggers the release of caspase-activating factors. Bid has also been reported to be cleaved by other proteases. Methodology/Principal Findings: To test the hypothesis that Bid is a central mediator of stress-induced apoptosis, we investigated the effects of a small molecule Bid inhibitor on stress-induced apoptosis, and generated HeLa cells deficient for Bid. Stable knockdown of bid lead to a pronounced resistance to Fas/CD95- and TRAIL-induced caspase activation and apoptosis, and significantly increased clonogenic survival. While Bid-deficient cells were equally sensitive to ER stress-induced apoptosis, they showed moderate, but significantly reduced levels of apoptosis, as well as increased clonogenic survival in response to the genotoxic drugs Etoposide, Oxaliplatin, and Doxorubicin. Similar effects were observed using the Bid inhibitor BI6C9. Interestingly, Bid-deficient cells were dramatically protected from apoptosis when subtoxic concentrations of ER stressors, Etoposide or Oxaliplatin were combined with subtoxic TRAIL concentrations. Conclusions/Significance: Our data demonstrate that Bid is central for death receptor-induced cell death and participates in anti-cancer drug-induced apoptosis in human cervical cancer HeLa cells. They also show that the synergistic effects of TRAIL in combination with either ER stressors or genotoxic anti-cancer drugs are nearly exclusively mediated via an increased activation of Bid-induced apoptosis signalling

The effect of skin fatty acids on Staphylococcus aureus

Staphylococcus aureus is a commensal of the human nose and skin. Human skin fatty acids, in particular cis-6-hexadecenoic acid (C-6-H), have high antistaphylococcal activity and can inhibit virulence determinant production. Here, we show that sub-MIC levels of C-6-H result in induction of increased resistance. The mechanism(s) of C-6-H activity was investigated by combined transcriptome and proteome analyses. Proteome analysis demonstrated a pleiotropic effect of C-6-H on virulence determinant production. In response to C-6-H, transcriptomics revealed altered expression of over 500 genes, involved in many aspects of virulence and cellular physiology. The expression of toxins (hla, hlb,hlgBC) was reduced, whereas that of host defence evasion components (cap, sspAB, katA) was increased. In particular, members of the SaeRS regulon had highly reduced expression, and the use of specific mutants revealed that the effect on toxin production is likely mediated via SaeRS

Associations between <i>paratuberculosis</i> status and milk production traits in Holstein cattle under consideration of interaction effects between test result and farm and lactation number

One of the most important factors for economic losses as result of infection with Mycobacterium avium ssp. paratuberculosis (MAP) is the decrease in milk yield. Different phenotyping methods for MAP lead to an inconsistent impact of MAP infection on milk parameters. It was the aim of the study to analyse the effect of the MAP status on milk yield, milk fat and milk protein (in kilograms), as well as on the calving interval and milk kilograms per day of life in German Holstein cattle. A dataset of 9&thinsp;367 faecal culture tested animals from 14 farms in Thuringia (Germany) were available. The MAP status of the animals affected milk yield and the milk parameters. Beside the effects of farm, lactation number and MAP status, the interaction between MAP status and farm on the milk parameters was significant. The latter result is a possible explanation for the inconsistent results from recent studies. For milk kg per day of life, the interaction between lactation number and MAP status also showed significance

Expression of Intermediate Filaments in the Developing Testis and Testicular Germ Cell Cancer

Cytokeratin and desmin expression have been associated with Sertoli cell maturity and the development of testicular germ cell cancer (TGCC). Thus, the present study aimed to characterize the expression of these intermediate filaments in normal testis development and TGCC. Cytokeratin and desmin were determined by immunohistochemistry and immunofluorescence in human fetal, and adult testis and tissue from patients with pre-invasive germ cell neoplasia in-situ (GCNIS) or invasive TGCC. Desmin was expressed in Sertoli cells of the human fetal testis, and the proportion of desmin expressing Sertoli cells was significantly reduced in the second trimester, compared with the first trimester (31.14% vs. 6.74%, p = 0.0016). Additionally, Desmin was expressed in the majority of Sertoli cells in the adult testis and TGCC samples. Cytokeratin was detected in Sertoli cells of human fetal testis but was not expressed in Sertoli cells of human adult testis. In patients with TGCC, cytokeratin was not expressed in Sertoli cells in tubules with active spermatogenesis but was detected in Sertoli cells in tubules containing GCNIS cells in patients with both pre-invasive and invasive TGCC. In conclusion, desmin was not associated with Sertoli cell maturation or progression to TGCC. However, cytokeratin appeared to be an indicator of impaired Sertoli cell maturation

Wingless-related integration site (WNT) signaling is activated during the inflammatory response upon cardiac surgery: A translational study

ObjectiveCardiac surgery and the use of cardiopulmonary bypass initiate a systemic inflammatory response. Wingless-related integration site (WNT) signaling is part of the innate immunity and has been attributed a major role in the regulation of inflammation. In preclinical research, WNT-5a may sustain an inflammatory response and cause endothelial dysfunction. Our aim was to investigate WNT signaling after cardiac surgery and its association with postoperative inflammation (Clinicaltrials.gov, NCT04058496).MethodsIn this prospective, single-center, observational study, 64 consecutive patients for coronary artery bypass grafting (CABG) ± valve surgery were assigned into three groups: off-pump CABG (n = 28), on-pump CABG (n = 16) and combined valve-CABG surgery (n = 20). Blood samples were acquired before surgery, at intensive care unit (ICU) admission and 4, 8, and 48 h thereafter. Plasma concentrations of WNT-5a and its antagonists Secreted frizzled-related protein 1 (sFRP-1), Secreted frizzled-related protein 5 (sFRP-5), and WNT inhibitory factor 1 (WIF-1) were determined by enzyme-linked immunosorbent assay. In addition, plasma concentrations of six inflammatory cytokines were measured by multiplex immunoassay. Parameters were analyzed for evolution of plasma concentration over time, interactions, intergroup differences, and association with clinical outcome parameters.ResultsAt baseline, WNT-5a, sFRP-1, and WIF-1 were present in a minimal concentration, while sFRP-5 was elevated. A higher baseline value of WNT-5a, sFRP-5, and WIF-1 resulted in higher subsequent values of the respective parameter. At ICU admission, WNT-5a and sFRP-5 reached their maximum and minimum value, respectively. WIF-1 decreased over time and was lowest 8 h after surgery. sFRP-1 changed minimally over time. While WNT-5a returned to the baseline within 48 h, sFRP-5 and WIF-1 did not reach their baseline value at 48 h. Of the investigated WNT system components, only WIF-1 partially reflected the severity of surgery. WNT-5a and WIF-1 had an impact on postoperative fluid balance and noradrenaline requirement.ConclusionWNT-5a, sFRP-5, and WIF-1 are part of the systemic inflammatory response after cardiac surgery. WNT-5a peaks immediately after cardiac surgery and returns to baseline within 48 h, presumably modulated by its antagonist sFRP-5. Based on this translational study, WNT-5a antagonism may be further investigated to assess potentially beneficial effects in patients with a dysregulated inflammation after cardiac surgery