Biosynthesis of UDP-α-N-Acetyl-d-mannosaminuronic Acid and CMP-ß-N-Acetyl-d-neuraminic Acid for the Capsular Polysaccharides of Campylobacter jejuni.

Campylobacter jejuni is a human pathogen and a leading cause of food poisoning in North America and Europe. The exterior surface of the bacterial cell wall is attached to a polymeric coat of sugar molecules known as the capsular polysaccharide (CPS) that helps protect the organism from the host immune response. The CPS is composed of a repeating sequence of common and unusual sugar residues. In the HS:11 serotype of C. jejuni, we identified two enzymes in the gene cluster for CPS formation that are utilized for the biosynthesis of UDP-α-N-acetyl-d-mannosaminuronic acid (UDP-ManNAcA). In the first step, UDP-α-N-acetyl-d-glucosamine (UDP-GlcNAc) is epimerized at C2 to form UDP-α-N-acetyl-d-mannosamine (UDP-ManNAc). This product is then oxidized by a NAD+-dependent C6-dehydrogenase to form UDP-ManNAcA. In the HS:6 serotype (C. jejuni strain 81116), we identified three enzymes that are required for the biosynthesis of CMP-ß-N-acetyl-d-neuraminic acid (CMP-Neu5Ac). In the first step, UDP-GlcNAc is epimerized at C2 and subsequently hydrolyzed to form N-acetyl-d-mannosamine (ManNAc) with the release of UDP. This product is then condensed with PEP by N-acetyl-d-neuraminate synthase to form N-acetyl-d-neuraminic acid (Neu5Ac). In the final step, CMP-N-acetyl-d-neuraminic acid synthase utilizes CTP to convert this product into CMP-Neu5Ac. A bioinformatic analysis of these five enzymes from C. jejuni serotypes HS:11 and HS:6 identified other bacterial species that can produce UDP-ManNAcA or CMP-Neu5Ac for CPS formation.

Metabolic Engineering of Escherichia coli for High-Titer Biosynthesis of 3'-Sialyllactose.

3'-Sialyllactose (3'-SL) is among the foremost and simplest sialylated breast milk oligosaccharides. In this study, an engineered Escherichia coli for high-titer 3'-SL biosynthesis was developed by introducing a multilevel metabolic engineering strategy, including (1) the introduction of precursor CMP-Neu5Ac synthesis pathway and high-performance α2,3-sialyltransferase (α2,3-SiaT) genes into strain BZ to achieve de novo synthesis of 3'-SL; (2) optimizing the expression of glmS-glmM-glmU involved in the UDP-GlcNAc and CMP-Neu5Ac synthesis pathways, and constructing a glutamine cycle system, balancing the precursor pools; (3) analysis of critical intermediates and inactivation of competitive pathway genes to redirect carbon flux to 3'-SL biosynthesis; and (4) enhanced catalytic performance of rate-limiting enzyme α2,3-SiaT by RBS screening, protein tag cloning. The final strain BZAPKA14 yielded 9.04 g/L 3'-SL in a shake flask. In a 3 L bioreactor, fed-batch fermentation generated 44.2 g/L 3'-SL, with an overall yield and lactose conversion of 0.53 g/(L h) and 0.55 mol 3'-SL/mol, respectively.

New Application of cycloSaligenyl Prodrugs Approach for the Delivery of Fosfoxacin Derivatives in Mycobacteria.

In this work, we implemented for the first time the cycloSaligenyl prodrug strategy to increase the bioavailability of fosmidomycin phosphate analogs in bacteria. Here, we report the synthesis of 34 cycloSaligenyl prodrugs of fosfoxacin and its derivatives. Among them, fifteen double prodrugs efficiently prevented the growth of the non-pathogenic, fast-growing Mycobacterium smegmatis.

Cell-free regeneration of ATP based on polyphosphate kinase 2 facilitates cytidine 5'-monophosphate production.

Cytidine 5'-monophosphate (5'-CMP), a key intermediate for the production of nucleotide derivatives, has been extensively used in food, agriculture, and medicine industries. Compared to RNA degradation and chemical synthesis, the biosynthesis of 5'-CMP has attracted wide attention due to its relatively low cost and eco-friendliness. In this study, we developed a cell-free regeneration of ATP based on polyphosphate kinase 2 (PPK2) to manufacture 5'-CMP from cytidine (CR). McPPK2 from Meiothermus cerbereus exhibited high specific activity (128.5 U/mg) and was used to accomplish ATP regeneration. McPPK2 and LhUCK (a uridine-cytidine kinase from Lactobacillus helveticus) were combined to convert CR to 5'-CMP. Further, the degradation of CR was inhibited by knocking out cdd from the Escherichia coli genome to enhance 5'-CMP production. Finally, the cell-free system based on ATP regeneration maximized the titer of 5'-CMP up to 143.5 mM. The wider applicability of this cell-free system was demonstrated in the synthesis of deoxycytidine 5'-monophosphate (5'-dCMP) from deoxycytidine (dCR) by incorporating McPPK2 and BsdCK (a deoxycytidine kinase from Bacillus subtilis). This study suggests that the cell-free regeneration of ATP based on PPK2 has the advantage of great flexibility for producing 5'-(d)CMP and other (deoxy)nucleotides.

The Full Cytosine-Cytosine Base Paring: Self-Assembly and Crystal Structure.

The synthesis of self-assembly systems that can mimic partial biological behaviours require ingenious and delicate design. For decades, scientists are committed to exploring new base pairing patterns using hydrogen bonds directed self-assembly of nucleotides. A fundamental question is the adaptive circumstance of the recognition between base pairs, namely, how solvent conditions affect the domain of base pairs. Towards this question, three nucleotide complexes based on 2'-deoxycytidine-5'-monophosphate (dCMP) and cytidine-5'-monophosphate (CMP) were synthesized in different solvents and pH values, and an unusual cytosine-cytosine base paring pattern (named full C : C base pairing) has been successfully obtained. Systematic single crystal analysis and 1 H NMR titration spectra have been performed to explore factors influencing the formation of base paring patterns. Moreover, supramolecular chirality of three complexes were studied using circular dichroism (CD) spectroscopy in solution and solid-state combined with crystal structure analysis.

Cardiovascular magnetic resonance imaging characteristics in patients with methamphetamine-associated cardiomyopathy.

BACKGROUND: Methamphetamine-associated cardiomyopathy (MA-CMP) is an increasingly recognised aetiology of cardiomyopathy. Cardiovascular magnetic resonance (CMR) is a specialised cardiac imaging modality commonly used in assessment of cardiomyopathy. We aimed to identify specific CMR features associated with MA-CMP. METHODS: A retrospective cohort study of CMR scans was performed in a single centre between January 2015 and December 2020. Thirty patients with MA-CMP who had undergone CMR were identified. MA-CMP was defined as those with a history of significant methamphetamine use hospitalised with acute decompensated heart failure (other causes of cardiomyopathy excluded). A retrospective analysis of index admission CMRs was performed. All studies were performed on a 1.5 T CMR scanner. RESULTS: The mean age of MA-CMP patients was 43.7 ± 7.5 years, and 86.7% were male. The mean left ventricular (LV) volume obtained in this cohort was consistent with severe LV dilatation (LV end-diastolic volume (334 ± 99 ml); LV end-systolic volume: 269 ± 98 ml), whilst the right ventricular (RV) volume indicated moderate-to-severe dilatation (RV end-diastolic volume: 272 ± 91 ml; RV end-systolic volume: 173 ± 82 ml). Mean LV ejection fraction (20.9 ± 9.2%) indicated severe LV dysfunction, with moderate-to-severe RV dysfunction also detected (RV ejection fraction: 29.4 ± 13.4%). 22 patients (73.3%) had myocardial late gadolinium enhancement (LGE), of which 59.1% were located in the mid-wall, with all of these involving the interventricular septum. 22.7% displayed localised regions of sub-endocardial LGE in a variety of locations, and 18.2% had transmural regions of LGE that were located in the inferior and inferolateral segments. 6 patients (20%) had intracardiac thrombus (4 LV, 2 both LV and RV). CONCLUSION: MA-CMP was associated with severe biventricular dilatation and dysfunction, with a high prevalence of intraventricular thrombus. This cohort study highlights that MA-CMP patients have a high prevalence of CMR findings.

Coordination Chemistry of Phosphate Groups in Systems Including Copper(II) Ions, Phosphoethanolamine and Pyrimidine Nucleotides.

The activity of phosphate groups of phosphoethanolamine and pyrimidine nucleotides (thymidine 5-monophosphate, cytidine 5-monophosphate and uridine 5'monophosphate) in the process of complexation metal ions in aqueous solution was studied. Using the potentiometric method with computer calculation of the data and spectroscopic methods such as UV-Vis, EPR, 13C and 31P NMR as well as FT-IR, the overall stability constants of the complexes as well as coordination modes were obtained. At lower pH, copper(II) ions are complexed only by phosphate groups, whereas the endocyclic nitrogen atom of nucleotides has been identified as a negative center interacting with the -NH3+ groups of phosphoethanolamine.

A comparison of isokinetic muscle strength in patients with chondromalacia patella: a cross-sectional study.

OBJECTIVE: Chondromalacia patella (CMP), which is one of the most common causes of anterior knee pain in young adults, is often accompanied by reflex inhibition of the quadriceps muscle. In this respect, a significant correlation between isokinetic parameters and knee muscle strengths would be expected. We hypothesized that an isokinetic dynamometer, which objectively evaluates muscle strength, may be an important guide in detecting muscle weakness in new-onset CMP and determining early treatment strategies. PATIENTS AND METHODS: A total of 113 participants (mean age 30.33 ± 6.96 years, min: 18, max: 44) were recruited and divided into two groups, thus a CMP group (n=48) and a control group (n=65). The symptom duration of the CMP group and the demographic characteristics of all participants were recorded. Knee flexion and extension muscle strengths were measured at angular velocities of 60°/s and 180°/s [Knee extension Peak Torque at 60°/s (PTE60), Knee flexion Peak Torque at 60°/s (PTF60), Knee extension Peak Torque at 180°/s (PTE180), Knee flexion Peak at 180°/s (PTF180) respectively] (five sets) using an isokinetic dynamometer. We also recorded the total work done in flexion and extension (TWDF and TWDE). A modified MRI staging system based on the Outerbridge arthroscopy system was used to stage CMP. Isokinetic dynamometric parameters were compared between CMP patients and healthy volunteers. RESULTS: 59 healthy volunteers (90.8%) were right-side dominant and 6 (9.2%) left-side dominant. 33 CMP patients (68.8%) were right-side dominant, and 15 (31.3%) left-side dominant. 20 (41.7%) CMP patients were classified as Stage 1, 20 (41.7%) as Stage 2, and 8 (16.7%) as Stage 3. All the PTF60, PTE60, PTF180, and PTE180 values were significantly lower in the CMP group than in healthy controls (all p < 0.05). CMP symptom duration ≥ 6 months was associated with significantly lower knee muscle strength than with symptom duration < 6 months (p < 0.05). Also, a statistically negative correlation was found between MRI stages and PTE60 values (p < 0.05). CONCLUSIONS: In conclusion, our findings show that the isokinetic dynamometer reveals muscle weakness in CMP patients, and weakness in isokinetic parameters was negatively correlated with symptom duration and MRI stages. Isokinetic knee muscle strength testing, together with other functional tools, enables the assessment of muscle weakness and early rehabilitation planning for patients with CMP.

Modeling road accident fatalities with underdispersion and zero-inflated counts.

In 2013, Thailand was ranked second in the world in road accident fatalities (RAFs), with 36.2 per 100,000 people. During the Songkran festival, which takes place during the traditional Thai New Year in April, the number of road traffic accidents (RTAs) and RAFs are markedly higher than on regular days, but few studies have investigated this issue as an effect of festivity. This study investigated the factors that contribute to RAFs using various count regression models. Data on 20,229 accidents in 2015 were collected from the Department of Disaster Prevention and Mitigation in Thailand. The Poisson and Conway-Maxwell-Poisson (CMP) distributions, and their zero-Inflated (ZI) versions were applied to fit the data. The results showed that RAFs in Thailand follow a count distribution with underdispersion and excessive zeros, which is rare. The ZICMP model marginally outperformed the CMP model, suggesting that having many zeros does not necessarily mean that the ZI model is required. The model choice depends on the question of interest, and a separate set of predictors highlights the distinct aspects of the data. Using ZICMP, road, weather, and environmental factors affected the differences in RAFs among all accidents, whereas month distinguished actual non-fatal accidents and crashes with or without deaths. As expected, actual non-fatal accidents were 2.37 times higher in April than in January. Using CMP, these variables were significant predictors of zeros and frequent deaths in each accident. The RAF average was surprisingly higher in other months than in January, except for April, which was unexpectedly lower. Thai authorities have invested considerable effort and resources to improve road safety during festival weeks to no avail. However, our study results indicate that people's risk perceptions and public awareness of RAFs are misleading. Therefore, nationwide road safety should instead be advocated by the authorities to raise society's awareness of everyday personal safety and the safety of others.

The retaining ß-Kdo glycosyltransferase WbbB uses a double-displacement mechanism with an intermediate adduct rearrangement step.

WbbB, a lipopolysaccharide O-antigen synthesis enzyme from Raoultella terrigena, contains an N-terminal glycosyltransferase domain with a highly modified architecture that adds a terminal ß-Kdo (3-deoxy-D-manno-oct-2-ulosonic acid) residue to the O-antigen saccharide, with retention of stereochemistry. We show, using mass spectrometry, that WbbB forms a covalent adduct between the catalytic nucleophile, Asp232, and Kdo. We also determine X-ray structures for the CMP-ß-Kdo donor complex, for Kdo-adducts with D232N and D232C WbbB variants, for a synthetic disaccharide acceptor complex, and for a ternary complex with both a Kdo-adduct and the acceptor. Together, these structures show that the enzyme-linked Asp232-Kdo adduct rotates to reposition the Kdo into a second sub-site, which then transfers Kdo to the acceptor. Retaining glycosyltransferases were thought to use only the front-side SNi substitution mechanism; here we show that retaining glycosyltransferases can also potentially use double-displacement mechanisms, but incorporating an additional catalytic subsite requires rearrangement of the protein's architecture.

Immune disguise: the mechanisms of Neu5Gc inducing autoimmune and transplant rejection.

Organ (stem cell) transplantation is the most effective treatment for advanced organ failure. Neu5Gc (N-hydroxyacetylneuraminic acid) is a pathogenic non-human sialic acid, which is very similar to the molecular structure of Neu5Ac (N-acetylneuraminic acid) in human body. Neu5Gc has the function of "immune disguise", which is the main obstacle to transplantation. Gene knockout such as cytidine monophosphate-N-acetylneuraminidase (CMAH) reduces donor antigenicity, making xenotransplantation from fiction to reality. Exploring the immune disguise event in this emerging field has become a hot topic in the research of transplantation immune tolerance mechanism.

More tools for our toolkit: The application of HEL-299 cells and dsRNA-nanoparticles to study human coronaviruses in vitro.

Human coronaviruses (HCoVs) are important human pathogens, as exemplified by the current SARS-CoV-2 pandemic. While the ability of type I interferons (IFNs) to limit coronavirus replication has been established, the ability of double-stranded (ds)RNA, a potent IFN inducer, to inhibit coronavirus replication when conjugated to a nanoparticle is largely unexplored. Additionally, the number of IFN competent cell lines that can be used to study coronaviruses in vitro are limited. In the present study, we show that poly inosinic: poly cytidylic acid (pIC), when conjugated to a phytoglycogen nanoparticle (pIC+NDX) is able to protect IFN-competent human lung fibroblasts (HEL-299 cells) from infection with different HCoV species. HEL-299 was found to be permissive to HCoV-229E, -OC43 and MERS-CoV-GFP but not to HCoV-NL63 or SARS-CoV-2. Further investigation revealed that HEL-299 does not contain the required ACE2 receptor to enable propagation of both HCoV-NL63 and SARS-CoV-2. Following 24h exposure, pIC+NDX was observed to stimulate a significant, prolonged increase in antiviral gene expression (IFNß, CXCL10 and ISG15) when compared to both NDX alone and pIC alone. This antiviral response translated into complete protection against virus production, for 4 days or 7 days post treatment with HCoV-229E or -OC43 when either pre-treated for 6h or 24h respectively. Moreover, the pIC+NDX combination also provided complete protection for 2d post infection when HEL-299 cells were infected with MERS-CoV-GFP following a 24h pretreatment with pIC+NDX. The significance of this study is two-fold. Firstly, it was revealed that HEL-299 cells can effectively be used as an IFN-competent model system for in vitro analysis of MERS-CoV. Secondly, pIC+NDX acts as a powerful inducer of type I IFNs in HEL-299, to levels that provide complete protection against coronavirus replication. This suggests an exciting and novel area of investigation for antiviral therapies that utilize innate immune stimulants. The results of this study will help to expand the range of available tools scientists have to investigate, and thus further understand, human coronaviruses.

Fluorine-functionalized conjugated microporous polymer as adsorbents for solid-phase extraction of nine perfluorinated alkyl substances.

Perfluorinated alkyl substances (PFASs) are persistent, toxic, ubiquitously distributed, and bioaccumulated substances, which have attracted increasing concern. To investigate the environmental effects of PFASs, there is a need to develop a sensitive, rapid, and efficient method for detecting trace level PFASs. In this study, a conjugated microporous polymer (CMP) with loading of fluorine, fabricated by Sonogashira-Hagihara cross-coupling, was exploited as a solid-phase extraction (SPE) adsorbent. The prepared fluorine-functionalized CMP (FCMP), which showed a large surface area of 1089 m2·g-1, high porosity, and good chemical stability, was used to extract PFASs from water samples. The adsorption mechanism was investigated using a sorption isotherm model, and the main interactions were fluorous and hydrophobic affinity. The FCMP-based SPE combined with high-performance liquid chromatography-tandem mass spectrometry achieved low limits of detection (0.19-0.97 ng·L-1), wide linear range (2-1600 ng·L-1), and good reproducibility (3.4%-12.9%) under the optimal conditions. Furthermore, the approach was utilized for the analysis of three water samples (snow, river water, and irrigation water) to evaluate its reliability, and satisfactory recovery (70.5%-127.5%) was obtained. Thus, FCMP was feasible SPE adsorbents for the selective extraction of PFASs.

mPR-Specific Actions Influence Maintenance of the Blood-Brain Barrier (BBB).

Cerebral cavernous malformations (CCMs) are characterized by abnormally dilated intracranial microvascular sinusoids that result in increased susceptibility to hemorrhagic stroke. It has been demonstrated that three CCM proteins (CCM1, CCM2, and CCM3) form the CCM signaling complex (CSC) to mediate angiogenic signaling. Disruption of the CSC will result in hemorrhagic CCMs, a consequence of compromised blood-brain barrier (BBB) integrity. Due to their characteristically incomplete penetrance, the majority of CCM mutation carriers (presumed CCM patients) are largely asymptomatic, but when symptoms occur, the disease has typically reached a clinical stage of focal hemorrhage with irreversible brain damage. We recently reported that the CSC couples both classic (nuclear; nPRs) and nonclassic (membrane; mPRs) progesterone (PRG)-receptors-mediated signaling within the CSC-mPRs-PRG (CmP) signaling network in nPR(-) breast cancer cells. In this report, we demonstrate that depletion of any of the three CCM genes or treatment with mPR-specific PRG actions (PRG/mifepristone) results in the disruption of the CmP signaling network, leading to increased permeability in the nPR(-) endothelial cells (ECs) monolayer in vitro. Finally, utilizing our in vivo hemizygous Ccm mutant mice models, we demonstrate that depletion of any of the three CCM genes, in combination with mPR-specific PRG actions, is also capable of leading to defective homeostasis of PRG in vivo and subsequent BBB disruption, allowing us to identify a specific panel of etiological blood biomarkers associated with BBB disruption. To our knowledge, this is the first report detailing the etiology to predict the occurrence of a disrupted BBB, an indication of early hemorrhagic events.

Recurrence Plot-based Classification of Ischemic and Dilated Cardiomyopathy Patients.

A large portion of the elderly population are affected by cardiovascular diseases. The early prognosis of cardiomyopathies is still a challenge. The aim of this study was to classify cardiomyopathy patients by their etiology in function of significant indexes extracted from the characterization of the recurrence plot of the systems involved. Thirty-nine cardiomyopathy patients (CMP) classified as ischemic (ICM - 24 patients) and dilated (DCM-15 patients) were considered. In addition, thirty-nine control subjects (CON) were used as reference. The beat-to-beat (BBI) time series, from the electrocardiographic signal, the systolic (SBP), and diastolic (DBP) time series, from the blood pressure signal, and the respiratory time (FLW) from the respiratory flow signal, were extracted. The recurrence plot from each signal considered were calculated and characterized by a total of 12 indexes. The best classifiers were used to build support vector machine models. The optimal model to classify ICM versus DCM patients achieved 92.3% accuracy, 95.8% sensitivity, and 86.6% specificity. When comparing CMP patients and CON subjects, the best model achieved 85.8% accuracy, 92.3% sensitivity, and 80.1% specificity. Our results suggest a more deterministic behavior in DCM patients. Clinical Relevance - This study explores the recurrence plot for the classification of ICM and DCM patients.

Enhanced Photocatalytic Efficiency in Visible-Light-Induced NADH Regeneration by Intramolecular Electron Transfer.

Inspired by natural photosynthesis, photocatalytic NADH regeneration has drawn increasing interest in the recent decade as it provides a perfect approach for NAD+ reduction into NADH, which can be further consumed by oxidordeuctase for enzymatic redox reactions. However, two issues still remain unsolved in this procedure. First, the photocatalytic efficiency in NAD+ hydrogenation requires further improvement. Second, the rhodium electron mediator [Cp*Rh(bpy)H2O]2+ (M), which is always required for selective 1,4-NADH regeneration, is difficult to recover because of its good solubility in aqueous solution. Given the high price of M, it is highly wasteful and inefficient if it only spends once. Here, we report a Cp*Rh(bpy)Cl implanted conjugated microporous polymer DTS/Rh@CMPs which can be employed as a highly effective visible light photocatalysts for in situ NADH regeneration without using additional M. In addition, the insertion of Rh complex into a polymer skeleton, as demonstrated in UV-vis, fluorescence, photocurrent and electrochemical impedance, dramatically improves the light absorption capacity and the electron separation and transfer efficiency. Compared with that of DTS@CMP-1 with M, an enhanced reaction yield of 33% was determined in DTS/Rh@CMP-1 suggesting that intramolecular electron transfer has a better activity than that of intermolecular electron transfer in photocatalytic NAD+ reduction. Moreover, as the Rh complex is rooted firmly in a polymer framework, negligible Rh loss and conversion decrease in NADH regeneration are observed. When the DTS/Rh@CMP-1 was coupled with yeast alcohol dehydrogenase (YADH, from Saccharomyces cerevisiae), 1.36 mM of methanol was accumulated, implying an excellent biocompatibility of DTS/Rh@CMP-1 and a high feasibility of photobiocatalysis for formaldehyde hydrogenation.

Adsorption separation of guanosine 5'-Monophosphate and cytidine 5'-Monophosphate by mixed-mode Resin HD-1: Experimental study and mathematical modeling.

The preparative separation of guanosine 5'-monophosphate (GMP) and cytidine 5'-monophosphate (CMP) on mixed-mode resin HD-1 was experimentally and theoretically investigated. The adsorption mechanisms of the two nucleotides were elucidated by analyzing adsorption equilibria and kinetics at different pH values. The adsorption dynamics of GMP and CMP in a fixed bed packed with resin HD-1 were studied. All nucleotide monovalent cations, zwitterions, and monovalent anions were adsorbed by the resin. Further, a general adsorption isotherm model was developed by considering the adsorption of different nucleotide species and the dissociation equilibrium behaviors of resin ligands. The model fit the adsorption isotherm data of GMP and CMP well. A modified liquid-film linear driving force model with the combined physical adsorption of nucleotides in different dissociation states and ion exchange of Na+ was established. The dissociation equilibria of resin ligands and nucleotides were considered. The model satisfactorily predicted the adsorption kinetic data at different pH values. The values of the efficient diffusion coefficients for GMP and CMP were not significantly influenced by the solution pH. A modified transport-diffusion model with pH gradient elution was proposed. The model accurately predicted the elution chromatographic peaks of GMP and CMP, as well as the pH at the outlet of the fixed bed packed with resin HD-1.

Findings and Outcome of Transcatheter Right Ventricular Endomyocardial Biopsy and Hemodynamic Assessment in Children with Suspected Myocarditis or Cardiomyopathy.

OBJECTIVE: The study objective is assessing findings and outcome in children with suspected cardiomyopathy (CMP) or myocarditis undergoing cardiac catheterization with transcatheter right ventricular endomyocardial biopsy (RV-EMB). METHODS: All consecutive children undergoing cardiac catheterization with RV-EMB for suspected CMP/myocarditis between 2002-2021 were analysed regarding clinical presentation, cardiac biomarkers, periprocedural management, hemodynamic, histological/immunohistological findings, and outcome. RESULTS: Eighty-five RV-EMBs were performed in 81 patients at a median age of 6.8 (IQR 9.9) years and a bodyweight of 20 (32.2) kg. Histological/immunohistological findings of RV-EMB revealed dilated CMP in 10 (12%), chronic myocarditis in 28 (33%), healing myocarditis in 5 (6%), acute myocarditis in 9 (11%), other heart muscle diseases in 23 (27%) (7 restrictive CMP, 5 hypertrophic CMP, 4 toxic/anthracycline-induced CMP, 4 endocardfibroelastosis, 1 arrhythmogenic right ventricular CMP, 1 laminin CMP, 1 haemangioma), no conclusive histology in 7 (8%), and normal histology in 3 (4%) patients. Median LVEDP was 17 mmHg (IQR 9), LAP 15 mmHg (10), and PVR 1.83 (1.87) Wood Units/m2. There were 3 major complications (3%), all patients recovered without any sequelae. At follow-up (median 1153, IQR 1799 days) 47 (59%) patients were alive, 11 (13%) dead, 15 (18%) underwent cardiac transplantation, and 8 (9%) were lost to follow-up. Death/cardiac transplantation occurred within 3 years from RV-EMB. All patients with an acute myocarditis survived. NT-pro-BNP, echo parameters, and invasive hemodynamics correlate independently with death/cardiac transplant. CONCLUSION: Hemodynamic invasive data and morphological findings in RV-EMB complete clinical diagnosis in children with suspected CMP/myocarditis and provide important information for further clinical management.

Individuals' beliefs about the biopsychosocial factors that contribute to their chronic musculoskeletal pain: protocol for a qualitative study in the UK.

INTRODUCTION: Chronic musculoskeletal pain (CMP) is described as pain that persists for longer than 3 months. At present, no research is available that understands why CMP develops and continues from the perspective of the individual. Research is needed to establish if there are any consistent biopsychosocial factors perceived as contributing to CMP and what informs such beliefs. Understanding individual beliefs will inform more effective communication between clinicians and patients about their CMP, as well as informing future research into the epidemiology of CMP. Interpretative phenomenological analysis will be used as a methodological framework as it explores how individuals make sense of their world through personal experiences and perceptions while preserving individual nuance. The aim of this study is to understand individuals' beliefs and perceptions about the biological, psychological and social factors, which contribute to the development and maintenance of their CMP. METHODS AND ANALYSIS: A qualitative study informed by the Consolidated Criteria for Reporting Qualitative Research using interpretative phenomenological analysis and semistructured interviews. A maximum variation purposive sample of 6-12 adults with CMP will be recruited from the general public in the UK. One semistructured interview will be conducted with each participant via an online video platform with interviews transcribed verbatim. The interview schedule (codesigned with expert patients and informed by existing evidence) identifies three domains of important questions; (1) patient beliefs on why they developed and continue to experience CMP; (2) the relationship between their biopsychosocial experiences and CMP; and (3) the origin of their beliefs. Strategies such as 'member checking' will be employed to ensure trustworthiness. ETHICS AND DISSEMINATION: Ethical approval was granted by the Research Ethics Office at the University of Birmingham (reference ERN_21-0813). Informed consent will be obtained from all participants. The study findings will be submitted for publication in a peer-reviewed journal and for presentation at conferences.

Carbon Monoxide Poisoning was Associated With Lifetime Suicidal Ideation: Evidence From A Population-Based Cross-Sectional Study in Hebei Province, China.

Objectives: We want to test the association between carbon monoxide poisoning (CMP) experiencing and lifetime suicidal ideation/suicide plan among community residents. Methods: This is a population-based cross-sectional study conducted among community residents in Hebei province, China. We analyzed a total of 21,376 valid questionnaires. CMP experience and lifetime suicidal ideation/suicide plan were assessed in this study. Logistic regression and false discovery rate correction were conducted to analyze the associations and correct the p values. Results: We found that CMP (OR = 2.56, p < 0.001, corrected-p = 0.001) was associated with lifetime suicidal ideation, and the other risk factors were female (OR = 0.53, p < 0.001, corrected-p = 0.001). The association between CMP and suicide plan was not supported after false discovery rate correction (OR = 2.15, p = 0.035, corrected-p = 0.385). For the CMP patients, experiencing ≥2 times CMP (OR = 2.76, p = 0.001, corrected-p = 0.011) was also in higher risk of lifetime suicidal ideation. The association between CMP times and lifetime suicidal plan was not supported after false discovery rate correction (OR = 4.95, p = 0.021, corrected-p = 0.231). Conclusion: CMP patients are in higher risk of lifetime suicidal ideation. For CMP patients, some strategies are needed to control their suicidal ideation.