Bovine Enteropathogenic Coronavirus: The Effect of the Host Cell and Trypsin Modification on the Virus Structure, Cytopathic Expression, and Infectivity.

Human rectal tumor-18 (HRT-18) cell clones 3F3, 3E3, D2, and 4B3 exhibited differences in cellular morphology in Giemsa-stained cultures and developing monolayers. Differences were evident in growth kinetics and plating efficiency of each clone. The clones produced colonies in soft agar, demonstrating anchorage independence. Cytopathic expression (CPE) including cytoplasmic vacuolization and cell fusion occurred in BCV-L9-infected clones 3F3, D2, and 3E3. Cell fusion was inapparent in clone 4B3. Bovine coronavirus strain L9 (BCV-L9) and 5 wild-type isolates replicated in HRT-18 cells, inducing cell fusion. Strain L9, exclusively, replicated in D2BFS cells, requiring trypsin to induce cell fusion. Strain L9 produced plaques in the HRT-18 clones, but the ease of plaque formation and plaque morphology was host cell dependent. Host cell-dependent plaque formation was demonstrated by wild-type BCV strains, and plaque morphology was strain dependent. The intensity of trypsin enhancement of CPE and plaque development depended on the virus strain and host cell. Trypsin greatly enhanced the infectivity of BCV-L9 in D2BFS and HRT-18 cells, and to lesser extents in clones 3F3, 3E3, and D2. Trypsin-enhanced infectivity was not detected in clone 4B3. The infectivity of strain LY-138 in HRT-18 cells was slightly enhanced by trypsin. Eleven structural proteins of BCV-L9 were detected by immunoblotting, including 185, 160, 140, 125, 110, 100, 52, 46, 37, 31-34, and 26-28 Kd species. Under reducing conditions 185, 140, and 100 Kd species migrated as 190, 65, and 95 Kd forms. Silver-stained proteins of 18, 20, and 23 Kd were reduced to a 20-23 Kd cluster. Host cell-dependent differences in the protein profile of L9 were detected. A 62 Kd protein was specific to L9 (D2BFS with trypsin). L9 (4B3) and L9 (D2) lacked the 46 and 110 Kd bands, respectively. Cleavage of the 185 Kd protein, an increase in the 100 Kd band, and conversion of the 31-34 Kd cluster to a 34 Kd band occurred when BCV-L9 was propagated in D2BFS cells with trypsin. Trypsin converted the reduced, 95 Kd protein found in L9 (HRT-18) to 90 Kd, and the 20-23 Kd cluster was altered to 19-23 Kd. In vitro trypsin treatment of L9 (3F3) resulted in cleavage of the 46 Kd molecule and the appearance of a 25 Kd species

The value of initial condition large ensembles to robust adaptation decision-making

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Mankin, J. S., Lehner, F., Coats, S., & McKinnon, K. A. The value of initial condition large ensembles to robust adaptation decision-making. Earth's Future, 8(10), (2020): e2012EF001610, doi:10.1029/2020EF001610.The origins of uncertainty in climate projections have major consequences for the scientific and policy decisions made in response to climate change. Internal climate variability, for example, is an inherent uncertainty in the climate system that is undersampled by the multimodel ensembles used in most climate impacts research. Because of this, decision makers are left with the question of whether the range of climate projections across models is due to structural model choices, thus requiring more scientific investment to constrain, or instead is a set of equally plausible outcomes consistent with the same warming world. Similarly, many questions faced by scientists require a clear separation of model uncertainty and that arising from internal variability. With this as motivation and the renewed attention to large ensembles given planning for Phase 7 of the Coupled Model Intercomparison Project (CMIP7), we illustrate the scientific and policy value of the attribution and quantification of uncertainty from initial condition large ensembles, particularly when analyzed in conjunction with multimodel ensembles. We focus on how large ensembles can support regional‐scale robust adaptation decision‐making in ways multimodel ensembles alone cannot. We also acknowledge several recently identified problems associated with large ensembles, namely, that they are (1) resource intensive, (2) redundant, and (3) biased. Despite these challenges, we show, using examples from hydroclimate, how large ensembles provide unique information for the scientific and policy communities and can be analyzed appropriately for regional‐scale climate impacts research to help inform risk management in a warming world.F. L. has been supported by the Swiss NSF (grant no. PZ00P2_174128), the NSF Division of Atmospheric and Geospace Sciences (grant no. AGS‐0856145, Amendment 87), and the Regional and Global Model Analysis (RGMA) component of the Earth and Environmental System Modeling Program of the U.S.Department of Energy’s Office of Biological & Environmental Research (BER) via NSF IA 1844590. This is SOEST publication no. 11115

Immunomodulator expression in trophoblasts from the feline immunodeficiency virus (FIV)-infected cat

<p>Abstract</p> <p>Background</p> <p>FIV infection frequently compromises pregnancy under experimental conditions and is accompanied by aberrant expression of some placental cytokines. Trophoblasts produce numerous immunomodulators that play a role in placental development and pregnancy maintenance. We hypothesized that FIV infection may cause dysregulation of trophoblast immunomodulator expression, and aberrant expression of these molecules may potentiate inflammation and compromise pregnancy. The purpose of this project was to evaluate the expression of representative pro-(TNF-α, IFN-γ, IL-1β, IL-2, IL-6, IL-12p35, IL-12p40, IL-18, and GM-CSF) and anti-inflammatory cytokines (IL-4, IL-5, and IL-10); CD134, a secondary co-stimulatory molecule expressed on activated T cells (FIV primary receptor); the chemokine receptor CXCR4 (FIV co-receptor); SDF-1α, the chemokine ligand to CXCR4; and FIV gag in trophoblasts from early-and late-term pregnancy.</p> <p>Methods</p> <p>We used an anti-cytokeratin antibody in immunohistochemistry to identify trophoblasts selectively, collected these cells using laser capture microdissection, and extracted total RNA from the captured cell populations. Real time, reverse transcription-PCR was used to quantify gene expression.</p> <p>Results</p> <p>We detected IL-4, IL-5, IL-6, IL-1β, IL-12p35, IL-12p40, and CXCR4 in trophoblasts from early-and late-term pregnancy. Expression of cytokines increased from early to late pregnancy in normal tissues. A clear, pro-inflammatory microenvironment was not evident in trophoblasts from FIV-infected queens at either stage of pregnancy. Reproductive failure was accompanied by down-regulation of both pro-and anti-inflammatory cytokines. CD134 was not detected in trophoblasts, and FIV gag was detected in only one of ten trophoblast specimens collected from FIV-infected queens.</p> <p>Conclusion</p> <p>Feline trophoblasts express an array of pro-and anti-inflammatory immunomodulators whose expression increases from early to late pregnancy in normal tissues. Non-viable pregnancies were associated with decreased expression of immunomodulators which regulate trophoblast invasion in other species. The detection of FIV RNA in trophoblasts was rare, suggesting that the high rate of reproductive failure in FIV-infected queens was not a direct result of viral replication in trophoblasts. The influence of placental immune cells on trophoblast function and pregnancy maintenance in the FIV-infected cat requires additional study.</p

Exercise-based cardiac rehabilitation for adults with heart failure

Background Chronic heart failure (HF) is a growing global health challenge. People with HF experience substantial burden that includes low exercise tolerance, poor health-related quality of life (HRQoL), increased risk of mortality and hospital admission, and high healthcare costs. The previous (2014) Cochrane systematic review reported that exercise-based cardiac rehabilitation (CR) compared to no exercise control shows improvement in HRQoL and hospital admission among people with HF, as well as possible reduction in mortality over the longer term, and that these reductions appear to be consistent across patient and programme characteristics. Limitations noted by the authors of this previous Cochrane Review include the following: (1) most trials were undertaken in patients with HF with reduced (< 45%) ejection fraction (HFrEF), and women, older people, and those with preserved (≥ 45%) ejection fraction HF (HFpEF) were under-represented; and (2) most trials were undertaken in the hospital/centre-based setting. Objectives To determine the effects of exercise-based cardiac rehabilitation on mortality, hospital admission, and health-related quality of life of people with heart failure. Search methods We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, Embase, and three other databases on 29 January 2018. We also checked the bibliographies of systematic reviews and two trial registers. Selection criteria We included randomised controlled trials that compared exercise-based CR interventions with six months’ or longer follow-up versus a no exercise control that could include usual medical care. The study population comprised adults (> 18 years) with evidence of HF - either HFrEF or HFpEF. Data collection and analysis Two review authors independently screened all identified references and rejected those that were clearly ineligible for inclusion in the review. We obtained full papers of potentially relevant trials. Two review authors independently extracted data from the included trials, assessed their risk of bias, and performed GRADE analyses. Main results We included 44 trials (5783 participants with HF) with a median of six months’ follow-up. For this latest update, we identified 11 new trials (N = 1040), in addition to the previously identified 33 trials. Although the evidence base includes predominantly patients with HFrEF with New York Heart Association classes II and III receiving centre-based exercise-based CR programmes, a growing body of studies include patients with HFpEF and are undertaken in a home-based setting. All included studies included a no formal exercise training intervention comparator. However, a wide range of comparators were seen across studies that included active intervention (i.e. education, psychological intervention) or usual medical care alone. The overall risk of bias of included trials was low or unclear, and we downgraded results using the GRADE tool for all but one outcome. Cardiac rehabilitation may make little or no difference in all-cause mortality over the short term (≤ one year of follow-up) (27 trials, 28 comparisons (2596 participants): intervention 67/1302 (5.1%) vs control 75/1294 (5.8%); risk ratio (RR) 0.89, 95% confidence interval (CI) 0.66 to 1.21; low-quality GRADE evidence) but may improve all-cause mortality in the long term (> 12 months follow up) (6 trials/comparisons (2845 participants): intervention 244/1418 (17.2%) vs control 280/1427 (19.6%) events): RR 0.88, 95% CI 0.75 to 1.02; high-quality evidence). Researchers provided no data on deaths due to HF. CR probably reduces overall hospital admissions in the short term (up to one year of follow-up) (21 trials, 21 comparisons (2182 participants): (intervention 180/1093 (16.5%) vs control 258/1089 (23.7%); RR 0.70, 95% CI 0.60 to 0.83; moderate-quality evidence, number needed to treat: 14) and may reduce HF-specific hospitalisation (14 trials, 15 comparisons (1114 participants): (intervention 40/562 (7.1%) vs control 61/552 (11.1%) RR 0.59, 95% CI 0.42 to 0.84; low-quality evidence, number needed to treat: 25). After CR, a clinically important improvement in shortterm disease-specific health-related quality of life may be evident (Minnesota Living With Heart Failure questionnaire - 17 trials, 18 comparisons (1995 participants): mean difference (MD) -7.11 points, 95% CI -10.49 to -3.73; low-quality evidence). Pooling across all studies, regardless of the HRQoL measure used, shows there may be clinically important improvement with exercise (26 trials, 29 comparisons (3833 participants); standardised mean difference (SMD) -0.60, 95% CI -0.82 to -0.39; I² = 87%; Chi² = 215.03; lowquality evidence). ExCR effects appeared to be consistent different models of ExCR delivery: centre vs. home-based, exercise dose, exercise only vs. comprehensive programmes, and aerobic training alone vs aerobic plus resistance programmes. Authors’ conclusions This updated Cochrane Review provides additional randomised evidence (11 trials) to support the conclusions of the previous version (2014) of this Cochane Review. Compared to no exercise control, CR appears to have no impact on mortality in the short term (< 12 months’ follow-up). Low- to moderate-quality evidence shows that CR probably reduces the risk of all-cause hospital admissions and may reduce HF-specific hospital admissions in the short term (up to 12 months). CR may confer a clinically important improvement in health-related quality of life, although we remain uncertain about this because the evidence is of low quality. Future ExCR trials need to continue to consider the recruitment of traditionally less represented HF patient groups including older, female, and HFpEF patients, and alternative CR delivery settings including home- and using technology-based programmes

Application of the speed-duration relationship to normalize the intensity of high-intensity interval training

The tolerable duration of continuous high-intensity exercise is determined by the hyperbolic Speed-tolerable duration (S-tLIM) relationship. However, application of the S-tLIM relationship to normalize the intensity of High-Intensity Interval Training (HIIT) has yet to be considered, with this the aim of present study. Subjects completed a ramp-incremental test, and series of 4 constant-speed tests to determine the S-tLIM relationship. A sub-group of subjects (n = 8) then repeated 4 min bouts of exercise at the speeds predicted to induce intolerance at 4 min (WR4), 6 min (WR6) and 8 min (WR8), interspersed with bouts of 4 min recovery, to the point of exercise intolerance (fixed WR HIIT) on different days, with the aim of establishing the work rate that could be sustained for 960 s (i.e. 4×4 min). A sub-group of subjects (n = 6) also completed 4 bouts of exercise interspersed with 4 min recovery, with each bout continued to the point of exercise intolerance (maximal HIIT) to determine the appropriate protocol for maximizing the amount of high-intensity work that can be completed during 4×4 min HIIT. For fixed WR HIIT tLIM of HIIT sessions was 399±81 s for WR4, 892±181 s for WR6 and 1517±346 s for WR8, with total exercise durations all significantly different from each other (P&#60;0.050). For maximal HIIT, there was no difference in tLIM of each of the 4 bouts (Bout 1: 229±27 s; Bout 2: 262±37 s; Bout 3: 235±49 s; Bout 4: 235±53 s; P&#62;0.050). However, there was significantly less high-intensity work completed during bouts 2 (153.5±40. 9 m), 3 (136.9±38.9 m), and 4 (136.7±39.3 m), compared with bout 1 (264.9±58.7 m; P&#62;0.050). These data establish that WR6 provides the appropriate work rate to normalize the intensity of HIIT between subjects. Maximal HIIT provides a protocol which allows the relative contribution of the work rate profile to physiological adaptations to be considered during alternative intensity-matched HIIT protocols

Effect of a Perioperative, Cardiac Output-Guided Hemodynamic Therapy Algorithm on Outcomes Following Major Gastrointestinal Surgery A Randomized Clinical Trial and Systematic Review

Importance: small trials suggest that postoperative outcomes may be improved by the use of cardiac output monitoring to guide administration of intravenous fluid and inotropic drugs as part of a hemodynamic therapy algorithm.Objective: to evaluate the clinical effectiveness of a perioperative, cardiac output–guided hemodynamic therapy algorithm.Design, setting, and participants: OPTIMISE was a pragmatic, multicenter, randomized, observer-blinded trial of 734 high-risk patients aged 50 years or older undergoing major gastrointestinal surgery at 17 acute care hospitals in the United Kingdom. An updated systematic review and meta-analysis were also conducted including randomized trials published from 1966 to February 2014.Interventions: patients were randomly assigned to a cardiac output–guided hemodynamic therapy algorithm for intravenous fluid and inotrope (dopexamine) infusion during and 6 hours following surgery (n=368) or to usual care (n=366).Main outcomes and measures: the primary outcome was a composite of predefined 30-day moderate or major complications and mortality. Secondary outcomes were morbidity on day 7; infection, critical care–free days, and all-cause mortality at 30 days; all-cause mortality at 180 days; and length of hospital stay.Results: baseline patient characteristics, clinical care, and volumes of intravenous fluid were similar between groups. Care was nonadherent to the allocated treatment for less than 10% of patients in each group. The primary outcome occurred in 36.6% of intervention and 43.4% of usual care participants (relative risk [RR], 0.84 [95% CI, 0.71-1.01]; absolute risk reduction, 6.8% [95% CI, ?0.3% to 13.9%]; P?=?.07). There was no significant difference between groups for any secondary outcomes. Five intervention patients (1.4%) experienced cardiovascular serious adverse events within 24 hours compared with none in the usual care group. Findings of the meta-analysis of 38 trials, including data from this study, suggest that the intervention is associated with fewer complications (intervention, 488/1548 [31.5%] vs control, 614/1476 [41.6%]; RR, 0.77 [95% CI, 0.71-0.83]) and a nonsignificant reduction in hospital, 28-day, or 30-day mortality (intervention, 159/3215 deaths [4.9%] vs control, 206/3160 deaths [6.5%]; RR, 0.82 [95% CI, 0.67-1.01]) and mortality at longest follow-up (intervention, 267/3215 deaths [8.3%] vs control, 327/3160 deaths [10.3%]; RR, 0.86 [95% CI, 0.74-1.00]).Conclusions and relevance: in a randomized trial of high-risk patients undergoing major gastrointestinal surgery, use of a cardiac output–guided hemodynamic therapy algorithm compared with usual care did not reduce a composite outcome of complications and 30-day mortality. However, inclusion of these data in an updated meta-analysis indicates that the intervention was associated with a reduction in complication rate