Small molecule WDR5 inhibitors down-regulate lncRNA expression.

WD repeat domain 5 (WDR5) plays an important role as a scaffold protein in both protein-protein and RNA-protein complexes involved in epigenetic gene regulation. In particular, some of these lncRNAs were reported to regulate the expression of genes in cis as well as themselves through binding WDR5. In this report, we investigate the two known binding sites of WDR5 in relation to lncRNA binding and expression. The WBM binding site mediates both protein-protein and lncRNA-protein interactions while the WIN site, which is on the opposite side of the protein, is only known to mediate protein-protein interactions. To dissect the function of different binding sites on WDR5, we characterized them with selective peptide ligands using fluorescence polarization and used these to demonstrate the selectivity of small molecule inhibitors of these two major binding sites. RNA immunoprecipitation experiments were performed to show that lncRNA-WDR5 complex formation could be interrupted using a WBM site inhibitor. Finally, we demonstrated that WDR5 regulated lncRNAs are down regulated with different sensitivity toward the corresponding inhibitors, demonstrating the potential of targeting lncRNA-protein interactions to reduce oncogenic lncRNA expression.

On-Resin Photochemical Decarboxylative Arylation of Peptides.

Here we describe the application of photochemical decarboxylative arylation as a late-stage functionalization reaction for peptides. The reaction uses redox-active esters of aspartic acid and glutamic acid on the solid phase to provide analogues of aromatic amino acids. By using aryl bromides as arylation reagents, a wide variety of amino acids can be accessed without having to synthesize them individually in solution. The reaction is compatible with proteinogenic amino acids and was used to perform a structure-activity relationship study of a PRMT5 binding peptide.

Activation of Adrenoceptor Alpha-2 (ADRA2A) Promotes Chemosensitization to Carboplatin in Ovarian Cancer Cell Lines.

Recurrence of ovarian cancer (OvCa) following surgery and standard carboplatin/paclitaxel first-line therapy signifies poor median progression-free survival (<24 months) in the majority of patients with OvCa. The current study utilized unbiased high-throughput screening (HTS) to evaluate an FDA-approved compound library for drugs that could be repurposed to improve OvCa sensitivity to carboplatin. The initial screen revealed six compounds with agonistic activity for the adrenoceptor alpha-2a (ADRA2A). These findings were validated in multiple OvCa cell lines (TYKnu, CAOV3, OVCAR8) using three ADRA2A agonists (xylazine, dexmedetomidine, and clonidine) and two independent viability assays. In all the experiments, these compounds enhanced the cytotoxicity of carboplatin treatment. Genetic overexpression of ADRA2A was also sufficient to reduce cell viability and increase carboplatin sensitivity. Taken together, these data indicate that ADRA2A activation may promote chemosensitivity in OvCa, which could be targeted by widely used medications currently indicated for other disease states.

A biofunctional review of C-reactive protein (CRP) as a mediator of inflammatory and immune responses: differentiating pentameric and modified CRP isoform effects.

C-reactive protein (CRP) is an acute phase, predominantly hepatically synthesized protein, secreted in response to cytokine signaling at sites of tissue injury or infection with the physiological function of acute pro-inflammatory response. Historically, CRP has been classified as a mediator of the innate immune system, acting as a pattern recognition receptor for phosphocholine-containing ligands. For decades, CRP was envisioned as a single, non-glycosylated, multi-subunit protein arranged non-covalently in cyclic symmetry around a central void. Over the past few years, however, CRP has been shown to exist in at least three distinct isoforms: 1.) a pentamer of five identical globular subunits (pCRP), 2.) a modified monomer (mCRP) resulting from a conformational change when subunits are dissociated from the pentamer, and 3.) a transitional isoform where the pentamer remains intact but is partially changed to express mCRP structural characteristics (referred to as pCRP* or mCRPm). The conversion of pCRP into mCRP can occur spontaneously and is observed under commonly used experimental conditions. In careful consideration of experimental design used in published reports of in vitro pro- and anti-inflammatory CRP bioactivities, we herein provide an interpretation of how distinctive CRP isoforms may have affected reported results. We argue that pro-inflammatory amplification mechanisms are consistent with the biofunction of mCRP, while weak anti-inflammatory mechanisms are consistent with pCRP. The interplay of each CRP isoform with specific immune cells (platelets, neutrophils, monocytes, endothelial cells, natural killer cells) and mechanisms of the innate immune system (complement), as well as differences in mCRP and pCRP ligand recognition and effector functions are discussed. This review will serve as a revised understanding of the structure-function relationship between CRP isoforms as related to inflammation and innate immunity mechanisms.

Spore-FP1 tuberculosis mucosal vaccine candidate is highly protective in guinea pigs but fails to improve on BCG-conferred protection in non-human primates.

Tuberculosis remains a major health threat globally and a more effective vaccine than the current Bacillus Calmette Guerin (BCG) is required, either to replace or boost it. The Spore-FP1 mucosal vaccine candidate is based on the fusion protein of Ag85B-Acr-HBHA/heparin-binding domain, adsorbed on the surface of inactivated Bacillus subtilis spores. The candidate conferred significant protection against Mycobacterium. tuberculosis challenge in naïve guinea pigs and markedly improved protection in the lungs and spleens of animals primed with BCG. We then immunized rhesus macaques with BCG intradermally, and subsequently boosted with one intradermal and one aerosol dose of Spore-FP1, prior to challenge with low dose aerosolized M. tuberculosis Erdman strain. Following vaccination, animals did not show any adverse reactions and displayed higher antigen specific cellular and antibody immune responses compared to BCG alone but this did not translate into significant improvement in disease pathology or bacterial burden in the organs.

iCatcher+: Robust and Automated Annotation of Infants' and Young Children's Gaze Behavior From Videos Collected in Laboratory, Field, and Online Studies.

Technological advances in psychological research have enabled large-scale studies of human behavior and streamlined pipelines for automatic processing of data. However, studies of infants and children have not fully reaped these benefits because the behaviors of interest, such as gaze duration and direction, still have to be extracted from video through a laborious process of manual annotation, even when these data are collected online. Recent advances in computer vision raise the possibility of automated annotation of these video data. In this article, we built on a system for automatic gaze annotation in young children, iCatcher, by engineering improvements and then training and testing the system (referred to hereafter as iCatcher+) on three data sets with substantial video and participant variability (214 videos collected in U.S. lab and field sites, 143 videos collected in Senegal field sites, and 265 videos collected via webcams in homes; participant age range = 4 months-3.5 years). When trained on each of these data sets, iCatcher+ performed with near human-level accuracy on held-out videos on distinguishing "LEFT" versus "RIGHT" and "ON" versus "OFF" looking behavior across all data sets. This high performance was achieved at the level of individual frames, experimental trials, and study videos; held across participant demographics (e.g., age, race/ethnicity), participant behavior (e.g., movement, head position), and video characteristics (e.g., luminance); and generalized to a fourth, entirely held-out online data set. We close by discussing next steps required to fully automate the life cycle of online infant and child behavioral studies, representing a key step toward enabling robust and high-throughput developmental research.

Rationally designed stapled peptides allosterically inhibit PTBP1-RNA-binding.

The diverse role of the splicing factor PTBP1 in human cells has been widely studied and was found to be a driver for several diseases. PTBP1 binds RNA through its RNA-recognition motifs which lack obvious pockets for inhibition. A unique transient helix has been described to be part of its first RNA-recognition motif and to be important for RNA binding. In this study, we further confirmed the role of this helix and envisioned its dynamic nature as a unique opportunity to develop stapled peptide inhibitors of PTBP1. The peptides were found to be able to inhibit RNA binding via fluorescence polarization assays and directly occupy the helix binding site as observed by protein crystallography. These cell-permeable inhibitors were validated in cellulo to alter the regulation of alternative splicing events regulated by PTBP1. Our study demonstrates transient secondary structures of a protein can be mimicked by stapled peptides to inhibit allosteric mechanisms.

Macrocyclic peptides as inhibitors of WDR5-lncRNA interactions.

WDR5 is an adaptor protein involved in the regulation of various epigenetic modifier complexes. Various inhibitors have been described but only as inhibitors of its protein-protein interactions. Here we describe peptidic macrocycles that act as inhibitors of the interaction between WDR5 and long non-coding RNAs. The findings provide a new strategy to modulate the biological function of WDR5 as an RNA binding epigenetic regulator.

Alkenyl Succinic Anhydride: The Question of Covalent Bonding and Chemistry Considerations for Better Sizing-Review.

Alkenyl Succinic Anhydride (ASA) is a sizing agent used in papermaking to increase the water repellency of paper. Almost 60 years after the introduction of the chemical in papermaking, scientists still have differing views on how ASA interacts with cellulose. Several experiments were conducted to bring more clarity to the ASA sizing mechanism, especially on the contentious question of ASA-cellulose covalent bonding or the esterification reaction between ASA and cellulose during papermaking. Herein, research papers and patents, including experiments and results, from the 1960s to 2020 were reviewed. Our investigation revealed that the ester bond formation between ASA and cellulose is insignificant and is not a prerequisite for sizing effectiveness; the main ASA-related material found in sized paper is hydrolyzed ASA or both hydrolyzed ASA and ASA salt. In addition, ASA emulsion stability and ASA emulsion retention are important for sizing efficiency improvement.

A translational repression reporter assay for the analysis of RNA-binding protein consensus sites.

RNA-binding proteins are essential regulators of RNA processing and function. Translational repression assays can be used to study how they interact with specific RNA sequences by insertion of such a consensus sequence into the 5' untranslated region of a reporter mRNA and measuring reporter protein translation. The straightforward set-up of these translational repression assays avoids the need for the isolation of the protein or the RNA providing speed, robustness and a low-cost method. Here, we report the optimization of the assay to function with linear RNA sequences instead of the previously reported hairpin type sequences to allow the study of a wider variety of RNA-binding proteins. Multiplication of a consensus sequence strongly improves the signal allowing analysis by both fluorescence intensity measurements and flow cytometry.

Standard of care for viral haemorrhagic fevers (VHFs): a systematic review of clinical management guidelines for high-priority VHFs.

The Sudan virus disease outbreak in Uganda in 2022 showed our vulnerability to viral haemorrhagic fevers (VHFs). Although there are regular outbreaks of VHFs with high morbidity and mortality, which disproportionally affect low-income settings, our understanding of how to treat them remains inadequate. In this systematic review, we aim to explore the availability, scope, standardisation, and quality of clinical management guidelines for VHFs. We identified 32 guidelines, 25 (78%) of which were low quality and did not have supporting evidence and eight (25%) of which had been produced or updated in the past 3 years. Guidance on supportive care and therapeutics had little detail and was sometimes contradictory. Guidelines based on uncertain evidence are a risk to patients, an ethical challenge for clinicians, and a challenge to implementing trials due to heterogeneous standards of care. We recommend a standard living guideline framework to improve the quality, scope, and applicability of guidelines. Furthermore, investments into trials should aim to identify optimal treatment strategies for VHFs and prioritise affordable and scalable interventions to improve outcomes globally.

Relationship between fitness performance and a newly developed continuous body composition score in U.S. adolescent boys.

OBJECTIVES: Body composition (BC) assessment typically requires the administration of a single test and can have different evaluation outcomes depending on the selected test and the specific population. The purpose of this study was twofold. Firstly, to develop and validate a novel continuous body composition (CBC) score using the continuous response model (CRM). Secondly, to examine the relationship between CBC scores and fitness performance. METHODS: Data from the 2012 NHANES National Youth Fitness Survey (NNYFS) were used and consisted of n=212 adolescent boys 12-15 years of age. CBC scale variables included body mass (BM), body mass index (BMI), arm circumference (AC), waist circumference (WC), calf circumference (CC), calf skinfold (CSF), triceps skinfold (TSF), and subscapular skinfold (SSF). Fitness performance variables included cardiorespiratory fitness (CRF, mL/kg/min), leg strength (LS, lb), modified pull-ups (MPU, #), grip strength (GS, kg), and plank (PL, sec). Samejima's CRM, factor analysis, convergent validity coefficients and score reliability were used to validate the CBC scale. Multinomial logistic regression and multiple linear regression were used to examine the relationship between CBC scores and fitness performance variables. RESULTS: Factor analysis of the CBC scale variables retained a single factor (loadings >0.81, 88% explained variance) with strong internal consistency (α=0.96). The CRM analysis indicated all CBC scale variables fit a unidimensional construct with adequate discrimination (as: 0.71-2.16) and difficulty (bs: -0.04-1.44). CBC scores (Mean=0, SD=1.00) displayed strong reliability (SEE.θ=0.22, r.θ=0.95) with lower values representing smaller-more-lean individuals and higher values representing larger-less-lean individuals. All fully adjusted regression models showed significant (ps<0.05) negative relationships between CBC scores and CRF, MPU, and PL and positive relationships between CBC scores and LS and GS. CONCLUSION: The CRM-derived CBC score is a novel measure of BC and found to be positively associated with strength performance and negatively associated with endurance performance in U.S. adolescent boys.

Relationships between physical activity and other health-related measures using state-based prevalence estimates.

Background: Both physical activity and muscle-strengthening activity have known relationships with other health-related variables such as alcohol and tobacco use, diet, and health-related quality of life (HRQOL). The purpose of this study was to explore and quantify the associations between physical activity measures and health-related variables at the higher state level. Methods: This cross-sectional study used data from the 2017 and 2019 Behavioral Risk Factor Surveillance System surveys. State-based prevalence (%) estimates were computed for meeting physical activity guidelines (PA), meeting muscle-strengthening activity guidelines (MS), both PA and MS (MB), drinking alcohol (D1), heavy alcohol drinking (HD), fruit consumption (F1), vegetable consumption (V1), good self-rated health (GH), overweight (OW), obesity (OB), current smoking (SN), and smokeless tobacco use (SL). Descriptive statistics, correlation coefficients, and data visualization methods were employed. Results: Strongest associations were seen between PA and F1 (2017: r=0.717 & 2019: r=0.695), MS and OB (2017: r=-0.781 & 2019: r=-0.599), PA and GH (2017: r=0.631 & 2019: r=0.649), PA and OB (2017: r=-0.645 & 2019: r=-0.763), and MB and SN (2017: r=-0.713 & 2019: r=-0.645). V1 was associated only with PA (2017: r=0.335 & 2019: r=0.357) whereas OW was not associated only with PA. Canonical correlation analysis showed the physical activity variables were directly related (r c=0.884, P<0.001) to the health variables. Conclusion: This study used high-level data to support the many known relationships between PA measures and health-related variables.

Development of Macrocyclic PRMT5-Adaptor Protein Interaction Inhibitors.

The PRMT5-MEP50 methyltransferase is a major target for anticancer drug discovery, and modulators of its interactions with different regulatory proteins are in high demand because they modulate PRMT5 substrate selectivity. We describe a strategy for the development of a PRMT5/adaptor protein PPI inhibitor, which includes the design and synthesis of macrocyclic peptides based on the motif for the interaction of PRMT5 with its adaptor protein RioK1. After the initial exploration of different macrocycle sizes and cyclization linkages, analysis of a peptide library identified hot spots for the variation of the amino acid structure. The incorporation of nonproteinogenic amino acids into the macrocyclic peptide led to a potent cyclic PRMT5 binding peptide (Ki = 66 nM), which selectively inhibits the interaction of PRMT5 with the adaptor proteins RioK1 and pICln (IC50 = 654 nM) but not with the alternative adaptor protein MEP50. The inhibitor is a promising tool for further biological investigation of this intriguing protein interface.

Preparing for pandemics: a systematic review of pandemic influenza clinical management guidelines.

BACKGROUND: The COVID-19 pandemic has highlighted the importance of evidence-based clinical decision-making. Clinical management guidelines (CMGs) may help reduce morbidity and mortality by improving the quality of clinical decisions. This systematic review aims to evaluate the availability, inclusivity, and quality of pandemic influenza CMGs, to identify gaps that can be addressed to strengthen pandemic preparedness in this area. METHODS: Ovid Medline, Ovid Embase, TRIP (Turning Research Into Practice), and Guideline Central were searched systematically from January 2008 to 23rd June 2022, complemented by a grey literature search till 16th June 2022. Pandemic influenza CMGs including supportive care or empirical treatment recommendations were included. Two reviewers independently extracted data from the included studies and assessed their quality using AGREE II (Appraisal of Guidelines for Research & Evaluation). The findings are presented narratively. RESULTS: Forty-eight CMGs were included. They were produced in high- (42%, 20/48), upper-middle- (40%, 19/48), and lower-middle (8%, 4/48) income countries, or by international organisations (10%, 5/48). Most CMGs (81%, 39/48) were over 5 years old. Guidelines included treatment recommendations for children (75%, 36/48), pregnant women (54%, 26/48), people with immunosuppression (33%, 16/48), and older adults (29%, 14/48). Many CMGs were of low quality (median overall score: 3 out of 7 (range 1-7). All recommended oseltamivir; recommendations for other neuraminidase inhibitors and supportive care were limited and at times contradictory. Only 56% (27/48) and 27% (13/48) addressed oxygen and fluid therapy, respectively. CONCLUSIONS: Our data highlights the limited availability of up-to-date pandemic influenza CMGs globally. Of those identified, many were limited in scope and quality and several lacked recommendations for specific at-risk populations. Recommendations on supportive care, the mainstay of treatment, were limited and heterogeneous. The most recent guideline highlighted that the evidence-base to support antiviral treatment recommendations is still limited. There is an urgent need for trials into treatment and supportive care strategies including for different risk populations. New evidence should be incorporated into globally accessible guidelines, to benefit patient outcomes. A 'living guideline' framework is recommended and further research into guideline implementation in different resourced settings, particularly low- and middle-income countries.

An evaluation of global Chikungunya clinical management guidelines: A systematic review.

Background: Chikungunya virus (CHIKV) has expanded its geographical reach in recent decades and is an emerging global health threat. CHIKV can cause significant morbidity and lead to chronic, debilitating arthritis/arthralgia in up to 40% of infected individuals. Prevention, early identification, and clinical management are key for improving outcomes. The aim of this review is to evaluate the quality, availability, inclusivity, and scope of evidence-based clinical management guidelines (CMG) for CHIKV globally. Methods: We conducted a systematic review. Six databases were searched from Jan 1, 1989, to 14 Oct 2021 and grey literature until Sept 16, 2021, for CHIKV guidelines providing supportive care and treatment recommendations. Quality was assessed using the appraisal of Guidelines for Research and Evaluation tool. Findings are presented in a narrative synthesis. PROSPERO registration: CRD42020167361. Findings: 28 CMGs were included; 54% (15/28) were produced more than 5 years ago, and most were of low-quality (median score 2 out of 7 (range 1-7)). There were variations in the CMGs' guidance on the management of different at-risk populations, long-term sequelae, and the prevention of disease transmission. While 54% (15/28) of CMGs recommended hospitalisation for severe cases, only 39% (11/28) provided guidance for severe disease management. Further, 46% (13/28) advocated for steroids in the chronic phase, but 18% (5/28) advised against its use. Interpretation: There was a lack of high-quality CMGs that provided supportive care and treatment guidance, which may impact patient care and outcomes. It is essential that existing guidelines are updated and adapted to provide detailed evidence-based treatment guidelines for different at-risk populations. This study also highlights a need for more research into the management of the acute and chronic phases of CHIKV infection to inform evidence-based care. Funding: The UK Foreign, Commonwealth and Development Office, Wellcome Trust [215091/Z/18/Z] and the Bill & Melinda Gates Foundation [OPP1209135].

Systemic maternal inflammation promotes ASD via IL-6 and IFN-γ.

Autism spectrum disorder (ASD) is a neurological disorder that manifests during early development, impacting individuals through their ways of communicating, social behaviors, and their ability to perform day-to-day activities. There have been different proposed mechanisms on how ASD precipitates within a patient, one of which being the impact cytokines have on fetal development once a mother's immune system has been activated (referred to as maternal immune activation, MIA). The occurrence of ASD has long been associated with elevated levels of several cytokines, including interleukin-6 (IL-6) and interferon gamma (IFN-γ). These proinflammatory cytokines can achieve high systemic levels in response to immune activating pathogens from various extrinsic sources. Transfer of cytokines such as IL-6 across the placental barrier allows accumulation in the fetus, potentially inducing neuroinflammation and consequently altering neurodevelopmental processes. Individuals who have been later diagnosed with ASD have been observed to have elevated levels of IL-6 and other proinflammatory cytokines during gestation. Moreover, the outcome of MIA has been associated with neurological effects such as impaired social interaction and an increase in repetitive behavior in animal models, supporting a mechanistic link between gestational inflammation and development of ASD-like characteristics. The present review attempts to provide a concise overview of the available preclinical and clinical data that suggest cross-talk between IL-6 and IFN-γ through both extrinsic and intrinsic factors as a central mechanism of MIA that may promote the development of ASD.

The Impact of an Education Intervention to Improve Blood Pressure Control among Black Non-Hispanic Patients and Hispanic Patients with Chronic Kidney Disease.

This project examined the effect of an educational intervention on blood pressure control among minority patients with chronic kidney disease (CKD). Adherence to a low-sodium diet is crucial for blood pressure control. It is also vital to assess food insecurity to improve diet adherence, especially among high-risk underrepresented populations. Participants were recruited from a public hospital renal clinic. Knowledge and food access was assessed using CKD and food insecurity questionnaires. Food frequency and 24-hour 3-day food diaries were completed and analyzed. Eighteen patients were enrolled (Black, non-Hispanic = 66.6%, Hispanic = 27.7%, uninsured = 33.3%, and Medicaid recipients = 27.7%). Eighty-nine percent of participants screened positive for food insecurity and received vouchers for healthy food from a food depository. Paired t tests showed statistically significant increase in knowledge (p < 0.00) and self-efficacy, and systolic blood pressure improved post-intervention. This study suggests that Black non-Hispanic and Hispanic patients with CKD have limited access to healthy food and consume higher sodium. Patient education, screening for food insecurity, and access to a food depository enhanced adherence to low sodium diet and improved blood pressure control.

Availability, scope and quality of monkeypox clinical management guidelines globally: a systematic review.

BACKGROUND: Monkeypox (MPX) is an important human Orthopoxvirus infection. There has been an increase in MPX cases and outbreaks in endemic and non-endemic regions in recent decades. We appraised the availability, scope, quality and inclusivity of clinical management guidelines for MPX globally. METHODS: For this systematic review, we searched six databases from inception until 14 October 2021, augmented by a grey literature search until 17 May 2022. MPX guidelines providing treatment and supportive care recommendations were included, with no exclusions for language. Two reviewers assessed the guidelines. Quality was assessed using the Appraisal of Guidelines for Research and Evaluation II tool. RESULTS: Of 2026 records screened, 14 guidelines were included. Overall, most guidelines were of low-quality with a median score of 2 out of 7 (range: 1-7), lacked detail and covered a narrow range of topics. Most guidelines focused on adults, five (36%) provided some advice for children, three (21%) for pregnant women and three (21%) for people living with HIV. Treatment guidance was mostly limited to advice on antivirals; seven guidelines advised cidofovir (four specified for severe MPX only); 29% (4/14) tecovirimat, and 7% (1/14) brincidofovir. Only one guideline provided recommendations on supportive care and treatment of complications. All guidelines recommended vaccination as post-exposure prophylaxis (PEP). Three guidelines advised on vaccinia immune globulin as PEP for severe cases in people with immunosuppression. CONCLUSION: Our results highlight a lack of evidence-based clinical management guidelines for MPX globally. There is a clear and urgent need for research into treatment and prophylaxis including for different risk populations. The current outbreak provides an opportunity to accelerate this research through coordinated high-quality studies. New evidence should be incorporated into globally accessible guidelines, to benefit patient and epidemic outcomes. A 'living guideline' framework is recommended. PROSPERO REGISTRATION NUMBER: CRD42020167361.