Thermal Adaptation of Westslope Cutthroat Trout

Populations of westslope cutthroat trout (Oncorhynchus clarkii lewisi), a State species of special concern, have declined throughout their native range. Genetic introgressions, mainly from rainbow trout (O. mykiss), but also from Yellowstone cutthroat trout (O. c. bouvieri), and habitat loss are believed to be the leading causes of this decline. Populations that remain are often small and isolated, thereby increasing their risk of inbreeding depression and extinction. Translocation projects may offer a solution by infusing new genetic material into populations and potentially increasing their probability of persistence. However, local adaptations must be considered when selecting a donor population. We investigated thermal adaptations of four wild populations of westslope cutthroat trout from the Missouri River drainage and one hatchery population from the Washoe Park Trout Hatchery, Anaconda, Montana. Two wild populations were deemed to be from warm streams and two from cold streams. Fish were spawned streamside and at the hatchery. The resulting embryos were placed in experimental systems at 8, 10, and 14 °C. Survival was monitored throughout incubation. Post-embryonic growth was measured 90 days after hatching. Relationships between population performance and natal stream thermal characteristics were examined for adaptive differences

Engineering key components in a synthetic eukaryotic signal transduction pathway

Signal transduction underlies how living organisms detect and respond to stimuli. A goal of synthetic biology is to rewire natural signal transduction systems. Bacteria, yeast, and plants sense environmental aspects through conserved histidine kinase (HK) signal transduction systems. HK protein components are typically comprised of multiple, relatively modular, and conserved domains. Phosphate transfer between these components may exhibit considerable cross talk between the otherwise apparently linear pathways, thereby establishing networks that integrate multiple signals. We show that sequence conservation and cross talk can extend across kingdoms and can be exploited to produce a synthetic plant signal transduction system. In response to HK cross talk, heterologously expressed bacterial response regulators, PhoB and OmpR, translocate to the nucleus on HK activation. Using this discovery, combined with modification of PhoB (PhoB-VP64), we produced a key component of a eukaryotic synthetic signal transduction pathway. In response to exogenous cytokinin, PhoB-VP64 translocates to the nucleus, binds a synthetic PlantPho promoter, and activates gene expression. These results show that conserved-signaling components can be used across kingdoms and adapted to produce synthetic eukaryotic signal transduction pathways

Investigation of Prader-Willi-like Phenotype using a Whole Genome Array

IntroductionPrader-Willi syndrome (PWS) is characterised byobesity, short stature, small hands and feet, neonatalhypotonia with difficulty in feeding at birth,hypogonadism and eye problems. At about two years ofage the feeding difficulties with poor suck are graduallyreplaced by hyperphagia and obsession with food,leading to the obesity. In addition to developmentaldelay which is manifested by short stature, small handsand feet, growth hormone deficiency andhypogenitalism/hypogonadism, there are alsobehavioural characteristics including learningdisabilities, temper tantrums, aggression, repetitivespeech, obsessive compulsive behaviour, sleep disorderand skin picking (Cassidy and Driscoll, 2009). Thisdisparate collection of symptoms led Holm et al (1993)to define the major and minor characteristics whichallowed a clinical diagnosis of this the most commongenetic form of obesity. Consensus diagnostic criteriawere defined and weighted scores in which the majorcriteria were awarded one point and the minor criteriahalf a point calculated. A score of 8 or more is clinicallydiagnostic for PWS.The majority of people with PWS have a paternallyderived deletion of approximately 5-7Mb in 15q11-q13,others have maternal disomy of chromosome 15(UPD15mat) and a minority have a defect of theimprinting centre located in exon 1 of the SNRPN genewhich leads to a maternal imprint on the paternallyderived chromosome. Any of these abnormalities willresult in loss of the paternal contribution to the Prader-Willi syndrome critical region (PWSCR), demonstratedby loss of a paternally derived unmethylated band at theimprinting centre and a lack of expression of the SNRPNgene. Although these do not differentiate between thedifferent genetic types of PWS they are diagnostic forthe syndrome (Cassidy and Driscoll, 2009; Ramsden etal, 2010; Zeschnigk et al, 1997).Within 15q11-q13 the complex imprintedSNURF/SNRPN gene hosts several untranslated snoRNAgenes located within intronic sequences. The finding ofa microdeletion involving SNORD116 in a boy with PWSled to the identification of this snoRNA as the candidategene for the syndrome (Sahoo et al, 2008).In the course of a large study of PWS in the UK(Whittington et al, 2001; Soni et al, 2007) three peoplewere identified who fulfilled the criteria for a clinicaldiagnosis of the syndrome but not the geneticlaboratory diagnostic criteria.The Affymetrix Cytogenetics Whole-Genome 2.7M arraywhile providing high resolution whole genome coveragereliably detects changes in copy number. Deletionsand/or duplications present in all three participants ifinvolved in annotated genes could potentiallycontribute to the Prader-Willi-like phenotype.Candidate genes can subsequently be evaluated toestimate their transcription levels and compared withthose shown by people with PWS and with unaffectedindividuals

Programmable Ligand Detection System in Plants through a Synthetic Signal Transduction Pathway

There is an unmet need to monitor human and natural environments for substances that are intentionally or unintentionally introduced. A long-sought goal is to adapt plants to sense and respond to specific substances for use as environmental monitors. Computationally re-designed periplasmic binding proteins (PBPs) provide a means to design highly sensitive and specific ligand sensing capabilities in receptors. Input from these proteins can be linked to gene expression through histidine kinase (HK) mediated signaling. Components of HK signaling systems are evolutionarily conserved between bacteria and plants. We previously reported that in response to cytokinin-mediated HK activation in plants, the bacterial response regulator PhoB translocates to the nucleus and activates transcription. Also, we previously described a plant visual response system, the de-greening circuit, a threshold sensitive reporter system that produces a visual response which is remotely detectable and quantifiable.We describe assembly and function of a complete synthetic signal transduction pathway in plants that links input from computationally re-designed PBPs to a visual response. To sense extracellular ligands, we targeted the computational re-designed PBPs to the apoplast. PBPs bind the ligand and develop affinity for the extracellular domain of a chemotactic protein, Trg. We experimentally developed Trg fusions proteins, which bind the ligand-PBP complex, and activate intracellular PhoR, the HK cognate of PhoB. We then adapted Trg-PhoR fusions for function in plants showing that in the presence of an external ligand PhoB translocates to the nucleus and activates transcription. We linked this input to the de-greening circuit creating a detector plant.Our system is modular and PBPs can theoretically be designed to bind most small molecules. Hence our system, with improvements, may allow plants to serve as a simple and inexpensive means to monitor human surroundings for substances such as pollutants, explosives, or chemical agents

Sources of stakeholder salience in the responsible investment movement: why do investors sign the Principles for Responsible Investment?

Since its inception in 2006, the United Nations-backed Principles for Responsible Investment (PRI) have grown to over 1300 signatories representing over $45 trillion. This growth is not slowing down. In this paper, we argue that there is a set of attributes which make the PRI salient as a stakeholder and its claim to sign the six PRI important to institutional investors. We use Mitchell et al.’s (Acad Manag Rev 22:853–886, 1997) theoretical framework of stakeholder salience, as extended by Gifford (J Bus Eth 92:79–97, 2010). We use as evidence confidential data from the annual survey of signatories carried out by the PRI in a 5-year period between 2007 and 2011. The findings highlight pragmatic and organizational legitimacy, normative and utilitarian power, and management values as the attributes that contribute most to the salience of the PRI as a stakeholder

The evolution of epigean and stygobitic species of Koonunga Sayce, 1907 (Syncarida: Anaspidacea) in Southern Australia, with the description of three new species

Three new species of Koonunga were discovered in surface and subterranean waters in southern Australia, and were defined using mtDNA analyses and morphology. The new species are: Koonunga hornei Leijs & King; K. tatiaraensis Leijs & King and K. allambiensis Leijs & King. Molecular clock analyses indicate that the divergence times of the species are older than the landscape that they currently inhabit. Different scenarios explaining this apparent discrepancy are discussed in the context of the palaeography of the area. A freshwater epigean origin for Koonunga is considered the most likely hypothesis, whereby some lineages made the transition to the subterranean environment within the last few million years influenced by significant climatic cooling/drying. We discuss the possibility that one stygobitic lineage secondarily regained some of its body pigmentation as adaptation to increased photic conditions after cave collapse and forming of cenotes during the last glacial maximum.Remko Leijs, Tessa Bradford, James G. Mitchell, William F. Humphreys, Steven J. B. Cooper, Peter Goonan, Rachael A. Kin

Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension

OBJECTIVE: To evaluate whether eculizumab helps patients with anti-acetylcholine receptor-positive (AChR+) refractory generalized myasthenia gravis (gMG) achieve the Myasthenia Gravis Foundation of America (MGFA) post-intervention status of minimal manifestations (MM), we assessed patients' status throughout REGAIN (Safety and Efficacy of Eculizumab in AChR+ Refractory Generalized Myasthenia Gravis) and its open-label extension. METHODS: Patients who completed the REGAIN randomized controlled trial and continued into the open-label extension were included in this tertiary endpoint analysis. Patients were assessed for the MGFA post-intervention status of improved, unchanged, worse, MM, and pharmacologic remission at defined time points during REGAIN and through week 130 of the open-label study. RESULTS: A total of 117 patients completed REGAIN and continued into the open-label study (eculizumab/eculizumab: 56; placebo/eculizumab: 61). At week 26 of REGAIN, more eculizumab-treated patients than placebo-treated patients achieved a status of improved (60.7% vs 41.7%) or MM (25.0% vs 13.3%; common OR: 2.3; 95% CI: 1.1-4.5). After 130 weeks of eculizumab treatment, 88.0% of patients achieved improved status and 57.3% of patients achieved MM status. The safety profile of eculizumab was consistent with its known profile and no new safety signals were detected. CONCLUSION: Eculizumab led to rapid and sustained achievement of MM in patients with AChR+ refractory gMG. These findings support the use of eculizumab in this previously difficult-to-treat patient population. CLINICALTRIALSGOV IDENTIFIER: REGAIN, NCT01997229; REGAIN open-label extension, NCT02301624. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that, after 26 weeks of eculizumab treatment, 25.0% of adults with AChR+ refractory gMG achieved MM, compared with 13.3% who received placebo

Minimal Symptom Expression' in Patients With Acetylcholine Receptor Antibody-Positive Refractory Generalized Myasthenia Gravis Treated With Eculizumab

The efficacy and tolerability of eculizumab were assessed in REGAIN, a 26-week, phase 3, randomized, double-blind, placebo-controlled study in anti-acetylcholine receptor antibody-positive (AChR+) refractory generalized myasthenia gravis (gMG), and its open-label extension

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570