Investigating diurnal changes in the chromatin organization of A. thaliana and the involvement of linker histone H1.

Light is not only a crucial source of energy for plants, but also delivers important information about the environment. It is perceived by a variety of photoreceptors that act to fine tune the plant cell metabolism and gene expression program. This enables adaptation to the environment. It remains unclear how light impacts gene expression, but it is thought to involve large-scale epigenetic and chromatin changes. During the diurnal rhythm genes are expressed in a coordinated manner and influence various physiological processes. This is thought to be controlled, at least partially, by changes in the 3D chromatin organization and by epigenetic modifications. A key regulator of genome packaging is the linker histone H1, which impacts molecular and spatial chromatin organization. Depletion of H1 in A. thaliana has been shown to cause aberrant root development in plants grown under different light regimes. However, it remains unclear how chromatin architecture is changing along the diurnal rhythm and what role H1 plays in this process. Our cytological analysis of chromatin features in A. thaliana revealed rhythmic abundance of H3K27me3, H3K4me3 and RNA Polymerase II S2P levels along the diurnal rhythm. Mutant analyses revealed that H1 contributes regulating the diurnal abundance of histone modifications and RNA Pol II activity, but only moderately influences Pol II S2P distributions. Major changes occur before light is turned on or off, suggesting a circadian entrainment to re-organize these chromatin features. Collectively, our results suggest a possible implication of H1 in regulating subtle, but significant chromatin dynamics at large scale, hence uncovering a novel function for linker histones in plants

A high-throughput screen identifying sequence and promiscuity characteristics of the loxP spacer region in Cre-mediated recombination

BACKGROUND: Cre-loxP recombination refers to the process of site-specific recombination mediated by two loxP sequences and the Cre recombinase protein. Transgenic experiments exploit integrative recombination, where a donor plasmid carrying a loxP site and DNA of interest integrate into a recipient loxP site in a target genome. Unfortunately, integrative recombination is highly inefficient because the insert is flanked by two loxP sites, which themselves become targets for Cre and lead to subsequent excision of the insert. A small number of mutations have been discovered in parts of the loxP sequence, specifically the spacer and inverted repeat segments, that increase the efficiency of integrative recombination. In this study we introduce a high-throughput in vitro assay to rapidly detect novel loxP spacer mutants and describe the sequence characteristics of successful recombinants. RESULTS: We created synthetic loxP oligonucleotides that contained a combination of inverted repeat mutations (the lox66 and lox71 mutations) and mutant spacer sequences, degenerate at 6 of the 8 positions. After in vitro Cre recombination, 3,124 recombinant clones were identified by sequencing. Included in this set were 31 unique, novel, self-recombining sequences. Using network visualization tools, we recognized 12 spacer sets with restricted promiscuity. We observed that increased guanine content at all spacer positions save for position 8 resulted in increased recombination. Interestingly, recombination between identical spacers was not preferred over non-identical spacers. We also identified a set of 16 pairs of loxP spacers that reacted at least twice with another spacer, but not themselves. Further, neither the wild-type P1 phage loxP sequence nor any of the known loxP spacer mutants appeared to be kinetically favoured by Cre recombinase. CONCLUSION: This study approached loxP spacer mutant screening in an unbiased manner, assuming nothing about candidate loxP sites save for the conserved 4 and 5 spacer positions. Candidate sites were free to recombine with any other sequence in the pool of all possible sites. The subset of loxP sites identified here are candidates for in vivo serial recombination as they have already demonstrated limited promiscuity with other loxP spacer and stability in the presence of Cre

Analise das diferenças e similaridades no conteúdo dos regimentos internos dos comitês de autoria das empresas de capital aberto

O estudo objetivou analisar o conteúdo dos regulamentos internos dos comitês de auditoria estatutários das empresas brasileiras de capital aberto listadas na B3. Foi utilizada uma pesquisa descritiva, documental e qualitativa com base em uma amostra de 111 empresas que divulgaram os Regimentos Internos dos Comitês de Auditoria Estatutários, no site da Comissão de Valores Mobiliários, no período de 01/01/2018 a 26/11/2021. Por fim, o objetivo do trabalho consiste em analisar o conteúdo disseminado pelas organizações nos respectivos relatórios, expondo quais conteúdos as empresas abordam ou deixam de abordar e ainda se seguem as normas e instruções das agências e instituições reguladoras. Em linhas gerais, pode-se concluir que as empresas nem sempre formalizam no Regimento Interno do Comitê de Auditoria o que é exigido pelas normas, ou seja, não mencionam as competências do comitê, a composição do comitê e nem tampouco deveres dos membros do Comitê. Além disso, nota-se que as empresas não seguem fidedignamente o conteúdo que deve mencionar em concordância com o que é pedido pelos órgãos regulamentadores

Direct transition from rapid-infusion originator to rapid-infusion biosimilar tumor necrosis factor inhibitor in children with inflammatory bowel disease: A case series.

PURPOSE: Biosimilar tumor necrosis factor inhibitors (b-TNFi) reduce healthcare costs and maintain equal efficacy when compared to their originator counterparts (o-TNFi). Current practice is to start patients on a slower standard infusion rate during the initial transition from an o-TNFi to a b-TNFi. There is a knowledge gap around switching from rapid originator infusion to rapid biosimilar infusion in the pediatric inflammatory bowel disease (IBD) population. SUMMARY: We present a case series of 8 pediatric patients with IBD who were switched from a rapid-infusion o-TNFi to a rapid-infusion b-TNFi from 2016 through 2022. Our primary interest was safety, which we evaluated based on the occurrence of infusion reactions or need for new premedications within the first 6 months of starting a b-TNFi. We also examined effectiveness through the incidence of IBD-related hospitalizations, TNFi failure, and need for co-medication or dose escalation over the same period. In our cohort, 4 patients had Crohns disease and 4 had ulcerative colitis. All patients were switched to a biosimilar for nonmedical reasons. During the follow-up period, no patients had infusion reactions necessitating new premedications, serious adverse events, or medication nonresponse. CONCLUSION: Patients who directly transitioned from a rapid-infusion o-TNFi to a rapid-infusion b-TNFi did not experience serious adverse events. Given the fiscal and patient experience advantages of rapid-rate infusions, larger studies are needed to consider a change in practice

Novel Therapies for Myocardial Irritability following Extreme Hydroxychloroquine Toxicity

Introduction. Hydroxychloroquine (HCQ) overdose is rare and potentially deadly when consumed in large doses. Management of severe HCQ toxicity is limited and infrequently reported. This report presents the case of a massive ingestion of HCQ. Case Report. A 23-year-old female presents following an intentional ingestion of approximately 40 g of HCQ. Within six hours after ingestion, she developed severe hemodynamic instability resulting from myocardial irritability with frequent ventricular ectopic activity leading to runs of polymorphic ventricular tachycardia (PMVT) and ventricular fibrillation (VF) requiring multiple defibrillations. Additional treatments included intravenous diazepam, epinephrine, norepinephrine, sodium bicarbonate, and magnesium sulfate. Despite the ongoing hemodynamic instability, the patient was also treated with Intralipid (ILE) and received hemodialysis. Improvements in her hemodynamics were observed after 18 hours. She survived her massive overdose of HCQ. Conclusion. HCQ poisoning is rare but serious because of its rapid progression to life-threatening symptoms. Hemodynamic support, gastric decontamination, electrolyte monitoring and replacement, and management of arrhythmias are the mainstays of treatment. The combined role of dialysis and ILE in the setting of massive HCQ overdose may improve outcomes

Improving ondansetron use and oral rehydration instructions for pediatric acute gastroenteritis.

In paediatric patients with acute gastroenteritis (AGE), ondansetron use decreases the need for intravenous fluids, reduces hospitalisations and shortens illness duration. Oral rehydration is also known to have excellent outcomes for mild to moderate dehydration secondary to AGE. Although these interventions are recommended in guidelines from international professional societies, baseline data at our clinic showed that <2% of these patients were offered ondansetron, and that few patients received appropriately detailed rehydration instructions. Therefore, we engaged residents and fellows as teachers and leaders in our university clinic's quality improvement programme to promote evidence-based practice for paediatric AGE. Our gap analysis identified opportunities for interventions including educating paediatricians and paediatrics residents on the safety and utility of the medication. We created standardised oral rehydration after-visit instructions and implemented a trainee-led educational approach that encouraged appropriate medication use. We used a follow-up survey to uncover provider concerns and tailor future interventions. The process metrics included: proportion of paediatric patients appropriately treated with ondansetron (goal of 80%), and proportion of patients given appropriate oral rehydration instructions. The outcome metric was 7-day representation rates. To achieve sustainability, we restructured our process to have senior residents take ownership of teaching and data collection. Trainee-driven interventions increased ondansetron prescription rates to a median of 66.6%. Patients prescribed ondansetron were less likely to represent to care, although representation rate was low overall. Postintervention data suggests that prescription rates decreased without continued interventions and additional systems redesign may help sustain impact.VoRSUNY DownstatePediatric GastroenterologyN/

Real‐world effectiveness of ustekinumab and vedolizumab in TNF‐exposed pediatric patients with ulcerative colitis

ObjectivesVedolizumab (VDZ) and ustekinumab (UST) are second-line treatments in pediatric patients with ulcerative colitis (UC) refractory to antitumor necrosis factor (anti-TNF) therapy. Pediatric studies comparing the effectiveness of these medications are lacking. Using a registry from ImproveCareNow (ICN), a global research network in pediatric inflammatory bowel disease, we compared the effectiveness of UST and VDZ in anti-TNF refractory UC.MethodsWe performed a propensity-score weighted regression analysis to compare corticosteroid-free clinical remission (CFCR) at 6 months from starting second-line therapy. Sensitivity analyses tested the robustness of our findings to different ways of handling missing outcome data. Secondary analyses evaluated alternative proxies of response and infection risk.ResultsOur cohort included 262 patients on VDZ and 74 patients on UST. At baseline, the two groups differed on their mean pediatric UC activity index (PUCAI) (p = 0.03) but were otherwise similar. At Month 6, 28.3% of patients on VDZ and 25.8% of those on UST achieved CFCR (p = 0.76). Our primary model showed no difference in CFCR (odds ratio: 0.81; 95% confidence interval [CI]: 0.41-1.59) (p = 0.54). The time to biologic discontinuation was similar in both groups (hazard ratio: 1.26; 95% CI: 0.76-2.08) (p = 0.36), with the reference group being VDZ, and we found no differences in clinical response, growth parameters, hospitalizations, surgeries, infections, or malignancy risk. Sensitivity analyses supported these findings of similar effectiveness.ConclusionsUST and VDZ are similarly effective for inducing clinical remission in anti-TNF refractory UC in pediatric patients. Providers should consider safety, tolerability, cost, and comorbidities when deciding between these therapies

Eight-Year Outcomes of Cardiosphere-Derived Cells in Single Ventricle Congenital Heart Disease

Background: Cardiosphere‐derived cell (CDC) infusion was associated with better clinical outcomes at 2 years in patients with single ventricle heart disease. The current study investigates time‐to‐event outcomes at 8 years. Methods and Results: This cohort enrolled patients with single ventricles who underwent stage 2 or stage 3 palliation from January 2011 to January 2015 at 8 centers in Japan. The primary outcomes were time‐dependent CDC treatment effects on death and late complications during 8 years of follow‐up, assessed by restricted mean survival time. Among 93 patients enrolled (mean age, 2.3±1.3 years; 56% men), 40 received CDC infusion. Overall survival for CDC‐treated versus control patients did not differ at 8 years (hazard ratio [HR], 0.60 [95% CI, 0.21–1.77]; P=0.35). Treatment effect had nonproportional hazards for death favoring CDCs at 4 years (restricted mean survival time difference +0.33 years [95% CI, 0.01–0.66]; P=0.043). In patients with heart failure with reduced ejection fraction, CDC treatment effect on survival was greater over 8 years (restricted mean survival time difference +1.58 years [95% CI, 0.05–3.12]; P=0.043). Compared with control participants, CDC‐treated patients showed lower incidences of late failure (HR, 0.45 [95% CI, 0.21–0.93]; P=0.027) and adverse events (subdistribution HR, 0.50 [95% CI, 0.27–0.94]; P=0.036) at 8 years. Conclusions: By 8 years, CDC infusion was associated with lower hazards of late failure and adverse events in single ventricle heart disease. CDC treatment effect on survival was notable by 4 years and showed a durable clinical benefit in patients with heart failure with reduced ejection fraction over 8 years. Registration: URL: https://www.clinicaltrials.gov; Unique identifiers: NCT01273857 and NCT01829750

From citizen science to jellyfish dispersion models and molecular studies : tracking the progress of jellyfish science in Malta (Central Mediterranean)

Following participation within the 1980”s FAO-mediated monitoring exercises of Pelagia noctiluca blooms within Maltese waters, little scientific effort was invested in studying the dynamics of jellyfish blooms within same waters and at developing management and public information strategies concerning the same blooms. A renewed scientific effort at studying such aspects within Maltese waters was registered from 2010 onwards, with the launch of the Spot the Jellyfish citizen science campaign (www.ioikids.net/jellyfish) which provided a user-friendly, multivalent and web-based through which maritime stakehold- ers and the public at large could submit their jellyfish records for Maltese waters. The web- based portal was also supported by other promotional initiatives in the field, such as the installation of seaside boards on beaches. Through this initiative, several previously-un- documented species of gelatinous plankton were recorded for the first time from the same waters, including Rhopilema nomadica, Aequorea forskalea, Porpita porpita, Discomedua lobata, Geryonia proboscidalis, Neotima lucullana, Physophora hydrostatica, Chrysaora hysoscella and Oceania armata. The maintenance of an updated jellyfish record database has been made possible through the conduction of such a citizen science initiative.peer-reviewe