Characterization of Microrna Expression Profiles and Role of Nodal-Related Genes in Zebrafish Ovarian Follicles

Zebrafish is a valuable model to study the biology of reproduction as the processes that regulate follicle development and oocyte maturation are conserved among vertebrates. In zebrafish, early vitellogenic (stage IIIa) ovarian follicles are maturationally incompetent while mid-late vitellogenic (stage IIIb) follicles are able to undergo oocyte maturation in response to maturation-inducing hormone signals. Signaling molecules derived from the ovary, such as microRNAs (miRNAs) and growth factors, are important in controlling ovarian function. To determine whether miRNAs may play a role in maturation competency acquisition, we characterized miRNA expression profiles in follicular cells isolated from stage IIIa and IIIb follicles. Bioinformatics analysis uncovered 214 known, 31 conserved novel and 44 novel miRNAs, of which 24 miRNAs were significantly regulated between stage IIIa and IIIb follicular cells. In addition, gene enrichment and pathway analyses of the predicted targets of the significantly regulated miRNAs supported the involvement of several key signaling pathways in regulating ovarian function. We then investigated the role of Nodal, a member of the transforming growth factor-β family, in regulating zebrafish ovarian function. We used real-time PCR to detect the zebrafish Nodal orthologs, nodal-related (ndr1) and ndr2 and found that they were expressed in ovarian follicles at all stages of development. We also detected the mRNAs for Nodal signaling components in follicular cells of vitellogenic follicles. Recombinant human Nodal activated Smad3, CREB, and ERK, and inhibited cell proliferation in ovarian follicular primary cell cultures. The mRNA levels of cyp17a1, hsd3b2 and paqr8 were increased in response to Nodal treatment. Subsequently, we used CRISPR/Cas9 technology to generate ndr1 and ndr2 null mutants, which caused severe defects in early development. To overcome this lethality in vivo, we developed a fluorescently-labeled, Doxycycline-inducible CRISPR-ON system that expresses single or multiplexed sgRNAs to knockout ndr1, ndr2, and ndr3. Activation of the system induced gene editing in the designated genomic loci. Our findings suggest that miRNAs and Nodal play a role in zebrafish follicles. The CRISPR-ON system will facilitate further investigating the roles of miRNAs and Nodal in adult zebrafish in vivo

Immunoglobulin, glucocorticoid, or combination therapy for multisystem inflammatory syndrome in children: a propensity-weighted cohort study.

BACKGROUND: Multisystem inflammatory syndrome in children (MIS-C), a hyperinflammatory condition associated with SARS-CoV-2 infection, has emerged as a serious illness in children worldwide. Immunoglobulin or glucocorticoids, or both, are currently recommended treatments. METHODS: The Best Available Treatment Study evaluated immunomodulatory treatments for MIS-C in an international observational cohort. Analysis of the first 614 patients was previously reported. In this propensity-weighted cohort study, clinical and outcome data from children with suspected or proven MIS-C were collected onto a web-based Research Electronic Data Capture database. After excluding neonates and incomplete or duplicate records, inverse probability weighting was used to compare primary treatments with intravenous immunoglobulin, intravenous immunoglobulin plus glucocorticoids, or glucocorticoids alone, using intravenous immunoglobulin as the reference treatment. Primary outcomes were a composite of inotropic or ventilator support from the second day after treatment initiation, or death, and time to improvement on an ordinal clinical severity scale. Secondary outcomes included treatment escalation, clinical deterioration, fever, and coronary artery aneurysm occurrence and resolution. This study is registered with the ISRCTN registry, ISRCTN69546370. FINDINGS: We enrolled 2101 children (aged 0 months to 19 years) with clinically diagnosed MIS-C from 39 countries between June 14, 2020, and April 25, 2022, and, following exclusions, 2009 patients were included for analysis (median age 8·0 years [IQR 4·2-11·4], 1191 [59·3%] male and 818 [40·7%] female, and 825 [41·1%] White). 680 (33·8%) patients received primary treatment with intravenous immunoglobulin, 698 (34·7%) with intravenous immunoglobulin plus glucocorticoids, 487 (24·2%) with glucocorticoids alone; 59 (2·9%) patients received other combinations, including biologicals, and 85 (4·2%) patients received no immunomodulators. There were no significant differences between treatments for primary outcomes for the 1586 patients with complete baseline and outcome data that were considered for primary analysis. Adjusted odds ratios for ventilation, inotropic support, or death were 1·09 (95% CI 0·75-1·58; corrected p value=1·00) for intravenous immunoglobulin plus glucocorticoids and 0·93 (0·58-1·47; corrected p value=1·00) for glucocorticoids alone, versus intravenous immunoglobulin alone. Adjusted average hazard ratios for time to improvement were 1·04 (95% CI 0·91-1·20; corrected p value=1·00) for intravenous immunoglobulin plus glucocorticoids, and 0·84 (0·70-1·00; corrected p value=0·22) for glucocorticoids alone, versus intravenous immunoglobulin alone. Treatment escalation was less frequent for intravenous immunoglobulin plus glucocorticoids (OR 0·15 [95% CI 0·11-0·20]; p<0·0001) and glucocorticoids alone (0·68 [0·50-0·93]; p=0·014) versus intravenous immunoglobulin alone. Persistent fever (from day 2 onward) was less common with intravenous immunoglobulin plus glucocorticoids compared with either intravenous immunoglobulin alone (OR 0·50 [95% CI 0·38-0·67]; p<0·0001) or glucocorticoids alone (0·63 [0·45-0·88]; p=0·0058). Coronary artery aneurysm occurrence and resolution did not differ significantly between treatment groups. INTERPRETATION: Recovery rates, including occurrence and resolution of coronary artery aneurysms, were similar for primary treatment with intravenous immunoglobulin when compared to glucocorticoids or intravenous immunoglobulin plus glucocorticoids. Initial treatment with glucocorticoids appears to be a safe alternative to immunoglobulin or combined therapy, and might be advantageous in view of the cost and limited availability of intravenous immunoglobulin in many countries. FUNDING: Imperial College London, the European Union's Horizon 2020, Wellcome Trust, the Medical Research Foundation, UK National Institute for Health and Care Research, and National Institutes of Health

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Analysis of Electricity and Water Consumption in Existing Mosque Buildings in the UAE

According to the World Economic Forum, the building sector is responsible for 40% of global energy consumption and 33% of greenhouse gas (GHG) emissions, and this is expected to increase due to population growth and the subsequent impact on the environment, economy and health. To tackle the problem, countries have set new construction codes, policies and regulations for the construction of new buildings in an effort to make them greener. However, there is a need to enhance the status of the existing buildings, especially mosques, as they are the main contributors to energy usage and water consumption in the United Arab Emirates (UAE). Therefore, this research seeks to fill this gap, aiming to evaluate the energy usage and water consumption practices employed in the existing mosque buildings within the UAE and to provide recommendations for improving the sustainability of mosques, with a focus on the environmental and economic pillars. The methodology relies mainly on data collected from 146 existing mosque buildings that have undergone energy saving audits across the UAE. Descriptive statistical analysis is performed to analyze the data from the period of 2018–2019 in order to determine the most significant factors related to energy inefficiency in existing mosque buildings in the UAE and to determine the most cost-effective and energy-saving corrective measures for energy and water conservation. The findings further enhance the standard of experience for mosque visitors (social aspect); reduce energy bill expenses, providing an acceptable return on investment from the proposed energy conservation measures for stakeholders (economic); and reduce the overall energy consumption, which can reduce the total CO2 emissions from mosque buildings (environmental)

Self-assembling peptide and nano-silver fluoride in remineralizing early enamel carious lesions: randomized controlled clinical trial

Abstract Background Nanoparticles and regenerative biomineralization are new caries prevention technologies. This study assessed the remineralizing effect of self-assembling peptide (P11-4), Nanosilver Fluoride (NSF) and sodium fluoride (NaF) on white spot lesions (WSLs) in permanent teeth. Methods Sixty six young adults with WSLs on buccal surfaces in permanent teeth and ICDAS code 1 or 2, were randomly assigned to one of three groups; P11-4, NSF or NaF. Assessment of ICDAS scores, lesion activity (Nyvad scores) and diagnodent readings of lesions were done at baseline and after 1, 3, 6 and 12 months of agents’ application. Comparisons between groups were made using chi squared test and comparison within groups were made using McNemar test. Multilevel binary logistic regression was used to assess the effect of agents on change of ICDAS scores after 3, 6 and 12 months (reduction versus no reduction). Results There were 147 teeth in 66 patients; mean ± SD age = 13.46 ± 4.31 years. There were significant differences in the change of ICDAS scores among the three groups after 3 and 6 months (p = 0.005). The reduction in ICDAS score increased steadily in all groups across time with the greatest increase in the P11-4 group: 54.5% after 12 months. Lesion activity (Nyvad scores) showed significant differences among the three groups with the greatest percentage of inactive cases in the P11-4 group. Multilevel binary logistic regression showed non-significant reduction of ICDAS in P11-4 and NSF varnishes compared to NaF varnish (AOR = 2.56, 95% CI: 0.58, 8.77 and AOR = 2.12, 95% CI: 0.59, 7.64 respectively). Conclusion P11-4 and NSF varnish reduced the ICDAS scores, caries activity and diagnodent readings of WSLs in permanent teeth. However, the change in ICDAS scores was not significantly different from NaF. Trial registration This trial was prospectively registered on the clinicaltrials.gov registry with ID: NCT04929509 on 18/6/2021

The Impact of Change Orders Caused by Legislative Changes on Program Management in the UAE Construction Industry

Program management is an important strategy for organizing and managing multiple interdependent construction projects to achieve strategic goals. However, when change orders occur, they can have a serious impact on the quality, time, cost of projects and, ultimately, affect the construction program. Furthermore, when change orders are caused particularly by legislative changes, such as environmental laws, taxes, tolls, safety codes, transportation, design or building codes, their impacts are unavoidable, yet can be managed through mitigation strategies. The existing literature only reports the implications of change orders on the project level and reports legislative changes as one of the contributing factors to change orders, but does not consider the implications on a program level. This study aims to close this knowledge gap by assessing the implications of change orders caused by legislative changes on program management in the construction industry during the construction phase, and explore what the possible mitigation strategies to manage change orders caused by legislative changes are. The objectives of the study include identifying the implications of change orders on construction projects in the UAE through a literature review using peer-reviewed journals and reliable industry sources. Additionally, we investigate the implications of change orders caused by legislative changes on construction programs through interviewing subject matter experts, evaluating the importance of the reported impacts, with possible mitigation strategies, through a structured questionnaire and Relative Importance Index (RII) and, finally, proposing a set of recommendations for key industry stakeholders. A mixed methods approach is adopted in this qualitative study, and the participants include clients, contractors and consultants from the construction industry, with a defined scope covering the construction stage only. The outcomes of the study can guide program managers, decision-makers and practitioners in the construction industry to successfully deliver all projects by directing proper resources to accommodate legislative changes

Minimizing treatment-induced emergence of antibiotic resistance in bacterial infections

Treatment of bacterial infections currently focuses on choosing an antibiotic that matches a pathogen’s susceptibility, with less attention paid to the risk that even susceptibility-matched treatments can fail as a result of resistance emerging in response to treatment. Combining whole-genome sequencing of 1113 pre- and posttreatment bacterial isolates with machine-learning analysis of 140,349 urinary tract infections and 7365 wound infections, we found that treatment-induced emergence of resistance could be predicted and minimized at the individual-patient level. Emergence of resistance was common and driven not by de novo resistance evolution but by rapid reinfection with a different strain resistant to the prescribed antibiotic. As most infections are seeded from a patient’s own microbiota, these resistance-gaining recurrences can be predicted using the patient’s past infection history and minimized by machine learning–personalized antibiotic recommendations, offering a means to reduce the emergence and spread of resistant pathogens

Additional file 1 of Risk factors associated with sickle cell retinopathy: findings from the Cooperative Study of Sickle Cell Disease

Additional file 1: Table S1. Characteristics of SCA and variant genotypes patients by PSCR Status