Evaluation of a casein glycomacropeptide-based protein substitute, in the dietary management of NTBC-induced tyrosinaemia in patients with alkaptonuria: A prospective open-label study.

BACKGROUND: 2-(2-Nitro-4-trifluoromethylbenzoyl)-1,3-cyclohexanedione (NTBC) treatment of alkaptonuria (AKU) leads to increased blood tyrosine levels, causing skin issues and potentially sight-threatening corneal keratopathy. Adherence to dietary management of NTBC-induced tyrosinemia, a low-protein diet with or without protein substitutes, can be difficult for patients. This 28-day interventional study evaluated a low tyrosine casein glycomacropeptide (cGMP) protein substitute (TYR sphere)®, a 20 g protein equivalent, cGMP-based protein substitute, in terms of adherence, palatability, usability, comparison to amino acid (AA)-based protein substitutes, gastrointestinal tolerance and metabolic control in adults with NTBC-induced tyrosinaemia. METHODS: Four adults (mean 61.1 years, range 53.3-69.3 years) with AKU and NTBC-induced tyrosinaemia were recruited from the United Kingdom National Alkaptonuria Centre (NAC). The cGMP protein substitute was prescribed based on individual nutritional requirements, replacing ≥1 AA-based protein substitute. Participants recorded product-related data in study diaries, using five-point Likert scales and daily and weekly logs. To determine metabolic control, prestudy blood tyrosine levels were compared to weekly blood spot tests during the study. RESULTS: Median cGMP protein substitute adherence was 98%. Most participants rated palatability and usability positively, and preferred cGMP protein substitute to AA-based products. There were no notable gastrointestinal changes, and metabolic control was maintained. CONCLUSIONS: cGMP protein substitute is a palatable and well-tolerated option in the dietary management of AKU patients with NTBC-induced tyrosinaemia.

Statin Usage in Peripheral Arterial Disease Patients.

OBJECTIVES: Atherosclerotic cardiovascular disease (ASCVD) events have been shown to occur at higher frequency in patients with peripheral arterial disease (PAD). In this study, our aim is to evaluate whether statin is being used appropriately in patients with PAD and also evaluate its usage with the number of vascular beds involved. MATERIALS AND METHODS: This retrospective cross-sectional study reviewed data of patients with a confirmed diagnosis of PAD based on invasive or noninvasive imaging. Demographic, clinical, laboratory, and treatment data collected were described using descriptive statistics. Multiple logistic regression analysis was conducted to determine the predictors for the prescription of statins (HIS). High-intensity statin therapy was defined as atorvastatin ≥40 mg per day, rosuvastatin ≥20 mg per day, or simvastatin ≥80 mg per day, according to American College of Cardiology (ACC)/American Heart Association (AHA) guidelines. RESULTS: We analyzed data from 166 patients who met the inclusion criteria. The mean age was 63.34 years. The most common comorbidity was diabetes mellitus (DM) (68.86%). Statins were used in 82% of patients, among whom only 39% were on high-intensity statins. Multiple logistic regression analysis revealed that patients with cerebrovascular disease (CVD) [odds ratio (OR) = 0.19, 95% confidence interval (CI) = 0.06-0.61, p = 0.005], on oral anticoagulants (OAC) (OR = 0.16, 95% CI = 0.04-0.62, p = 0.008) and on dual antiplatelet therapy (DAPT) (OR = 0.20, 95% CI = 0.08-0.47, p < 0.000) had lower odds of receiving lower extremity revascularization (LIS) therapy. CONCLUSION: Despite having a high risk of future adverse cardiac events, patients with PAD are less likely to receive appropriate statin therapy. Involvement of more vascular beds was associated with higher chances of initiating high-intensity statin.

Impaired adhesion of neutrophils expressing Slc44a2/HNA-3b to VWF protects against NETosis under venous shear rates.

Genome-wide association studies linked expression of the human neutrophil antigen 3b (HNA-3b) epitope on the Slc44a2 protein with a 30% decreased risk of venous thrombosis (VT) in humans. Slc44a2 is a ubiquitous transmembrane protein identified as a receptor for von Willebrand factor (VWF). To explain the link between Slc44a2 and VT, we wanted to determine how Slc44a2 expressing either HNA-3a or HNA-3b on neutrophils could modulate their adhesion and activation on VWF under flow. Transfected HEK293T cells or neutrophils homozygous for the HNA-3a- or HNA-3b-coding allele were purified from healthy donors and perfused in flow chambers coated with VWF at venous shear rates (100 s-1). HNA-3a expression was required for Slc44a2-mediated neutrophil adhesion to VWF at 100 s-1. This adhesion could occur independently of ß2 integrin and was enhanced when neutrophils were preactivated with lipopolysaccharide. Moreover, specific shear conditions with high neutrophil concentration could act as a "second hit," inducing the formation of neutrophil extracellular traps. Neutrophil mobilization was also measured by intravital microscopy in venules from SLC44A2-knockout and wild-type mice after histamine-induced endothelial degranulation. Mice lacking Slc44a2 showed a massive reduction in neutrophil recruitment in inflamed mesenteric venules. Our results show that Slc44a2/HNA-3a is important for the adhesion and activation of neutrophils in veins under inflammation and when submitted to specific shears. The fact that neutrophils expressing Slc44a2/HNA-3b have a different response on VWF in the conditions tested could thus explain the association between HNA-3b and a reduced risk for VT in humans.

CATS: Cas9-assisted tag switching. A high-throughput method for exchanging genomic peptide tags in yeast.

BACKGROUND: The creation of arrays of yeast strains each encoding a different protein with constant tags is a powerful method for understanding how genes and their proteins control cell function. As genetic tools become more sophisticated there is a need to create custom libraries encoding proteins fused with specialised tags to query gene function. These include protein tags that enable a multitude of added functionality, such as conditional degradation, fluorescent labelling, relocalization or activation and also DNA and RNA tags that enable barcoding of genes or their mRNA products. Tools for making new libraries or modifying existing ones are becoming available, but are often limited by the number of strains they can be realistically applied to or by the need for a particular starting library. RESULTS: We present a new recombination-based method, CATS - Cas9-Assisted Tag Switching, that switches tags in any existing library of yeast strains. This method employs the reprogrammable RNA guided nuclease, Cas9, to both introduce endogenous double strand breaks into the genome as well as liberating a linear DNA template molecule from a plasmid. It exploits the relatively high efficiency of homologous recombination in budding yeast compared with non-homologous end joining. CONCLUSIONS: The method takes less than 2 weeks, is cost effective and can simultaneously introduce multiple genetic changes, thus providing a rapid, genome-wide approach to genetic modification.

Natural Product-Derived Chiral Pyrrolidine-2,5-diones, Their Molecular Structures and Conversion to Pharmacologically Important Skeletons.

The versatility of the natural products (2S,3S)- and (2S,3R)-3-hydroxy-5-oxotetrahydrofuran-2,3-dicarboxylic acids (1 and 2), isolated in large amounts from tropical plant sources, has been demonstrated by the construction of 3-substituted and 3,4-disubstituted chiral pyrrolidine-2,5-diones. The absolute configurations of chiral pyrrolidine-2,5-diones have been ascertained using chiroptical spectroscopic methods and/or single-crystal XRD data. A combination of different reaction strategies delivering a diverse matrix of fused heterocyclic ring systems is presented. The pyrrolo[2,1-a]isoquinoline alkaloid (+)-crispine A possesses a wide range of pharmacological activities including antidepressant, antiplatelet, antileukemic, and anticancer activities. The analogues of indolizino[8,7-b]indole alkaloids (+)- and (-)-harmicine show strong antileishmanial, antinociceptive, PDE5-inhibitory, antimalarial, and antiviral activities. The bicyclic furo[2,3-b]pyrrolo skeleton is present in many natural products. Thus, the uniqueness of relatively cheap, naturally occurring chiral 2-hydroxycitric acid lactones as chirons has been demonstrated by the construction of some important molecular skeletons that are otherwise difficult to synthesize.

Tissue factor-positive neutrophils bind to injured endothelial wall and initiate thrombus formation.

For a long time, blood coagulation and innate immunity have been viewed as interrelated responses. Recently, the presence of leukocytes at the sites of vessel injury has been described. Here we analyzed interaction of neutrophils, monocytes, and platelets in thrombus formation after a laser-induced injury in vivo. Neutrophils immediately adhered to injured vessels, preceding platelets, by binding to the activated endothelium via leukocyte function antigen-1-ICAM-1 interactions. Monocytes rolled on a thrombus 3 to 5 minutes postinjury. The kinetics of thrombus formation and fibrin generation were drastically reduced in low tissue factor (TF) mice whereas the absence of factor XII had no effect. In vitro, TF was detected in neutrophils. In vivo, the inhibition of neutrophil binding to the vessel wall reduced the presence of TF and diminished the generation of fibrin and platelet accumulation. Injection of wild-type neutrophils into low TF mice partially restored the activation of the blood coagulation cascade and accumulation of platelets. Our results show that the interaction of neutrophils with endothelial cells is a critical step preceding platelet accumulation for initiating arterial thrombosis in injured vessels. Targeting neutrophils interacting with endothelial cells may constitute an efficient strategy to reduce thrombosis.

Extracellular DNA traps are associated with the pathogenesis of TRALI in humans and mice.

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related death. The biologic processes contributing to TRALI are poorly understood. All blood products can cause TRALI, and no specific treatment is available. A "2-event model" has been proposed as the trigger. The first event may include surgery, trauma, or infection; the second involves the transfusion of antileukocyte antibodies or bioactive lipids within the blood product. Together, these events induce neutrophil activation in the lungs, causing endothelial damage and capillary leakage. Neutrophils, in response to pathogens or under stress, can release their chromatin coated with granule contents, thus forming neutrophil extracellular traps (NETs). Although protective against infection, these NETs are injurious to tissue. Here we show that NET biomarkers are present in TRALI patients' blood and that NETs are produced in vitro by primed human neutrophils when challenged with anti-HNA-3a antibodies previously implicated in TRALI. NETs are found in alveoli of mice experiencing antibody-mediated TRALI. DNase 1 inhalation prevents their alveolar accumulation and improves arterial oxygen saturation even when administered 90 minutes after TRALI onset. We suggest that NETs form in the lungs during TRALI, contribute to the disease process, and thus could be targeted to prevent or treat TRALI.

Percutaneous ultrasound-guided anchor wire placement aids in the intraoperative localization of nonpalpable, superficial foreign bodies and abscesses in dogs.

OBJECTIVE: To describe and evaluate the use of preoperative percutaneous ultrasound-guided anchor wire placement to aid intraoperative localization of superficial foreign bodies and abscesses in dogs. ANIMALS: 11 dogs. CLINICAL PRESENTATION: In a retrospective observational study, the medical records of dogs that underwent surgical exploration of superficial abscesses, guided by anchor wire, between 2018 and 2023 were reviewed for clinical and histopathological findings and complications. Owners or veterinarians were contacted to collect long-term follow-up information. RESULTS: 11 dogs were included. Superficial swelling was the most common clinical presentation. Computed tomography and ultrasound revealed an abscess cavity and suspected foreign body in 9 dogs and an abscess cavity without evidence of a foreign body in 2 dogs. Anchor wires were placed in close proximity to the foreign body or inside the abscess. All documented foreign bodies were successfully located and retrieved. Two dogs suffered minor postoperative complications. No major intra- or postoperative complications were documented. One dog displayed recurrence of clinical signs, but no further surgical management was required. CLINICAL RELEVANCE: Preoperative percutaneous placement of an anchor wire via ultrasound guidance was successful in aiding intraoperative localization of nonpalpable abscesses and retrieval of foreign bodies. This technique may decrease surgical time, minimize the surgical approach required, and increase the likelihood of successful localization.

Distal partial ulnectomy with ulnar styloid process excision for management of an aneurysmal bone cyst in a cat.

Case summary: An 8-year-old male neutered domestic shorthair cat was presented with an acute onset of left thoracic limb lameness and a firm swelling on the lateral aspect of the left distal antebrachium. A CT scan of the left thoracic limb revealed an expansile osteolytic cystic bone lesion centred at the distal left ulnar metaphysis. Cytology from fine-needle aspiration was not consistent with neoplasia. The CT features and the cytology results were suggestive of a bone cyst. A distal partial ulnectomy with ulnar styloid process excision was performed as the biopsy method and as the treatment approach. Histopathology results were consistent with an aneurysmal bone cyst (ABC). Carpal instability was not detected after distal partial ulnectomy; therefore, a stabilisation method was not required. Limb function was excellent after surgery, with no lameness and no recurrence detected by the owner at 2, 6 and 24 weeks postoperatively. The veterinary examination at 12 weeks postoperatively confirmed the owner's outcome assessment. Relevance and novel information: To the authors' knowledge, this is the first report describing a distal partial ulnectomy with ulnar styloid process excision in a cat. Despite disrupting the short ulnar collateral ligament, this technique provided excellent short-term limb function with no need for carpal joint stabilisation. This technique allowed for the complete excision of a distal ulnar ABC and avoided cyst debridement, which could be associated with haemorrhage, recurrence and malignant transformation. Distal partial ulnectomy should be considered for distal ulnar bone lesions in cats.

Exploring the Efficacy of Sotagliflozin on Heart and Kidney Health in Diabetic Patients: A Comprehensive Meta-Analysis.

Evidence for reducing cardiovascular and renal events with sotagliflozin is uncertain among type 2 diabetes mellitus (T2DM) patients. To gather more evidence, this meta-analysis assesses the beneficial effects of sotagliflozin, a dual sodium-glucose cotransporter 1 and 2 inhibitor, in reducing the cardiovascular and renal events in diabetic patients with or without chronic kidney disease (CKD). Scopus, Google Scholar, Cochrane Central Register of Controlled Trials (CENTRAL), and PubMed were the databases used to search. The studies published from January 1, 2018, to January 30, 2022, were considered. The eligibility of studies was assessed independently. The data were collected in a modified Cochrane data extraction form. The included studies' quality was assessed with the Cochrane risk-of-bias tool. The quality of evidence for renal and cardiovascular outcomes was evaluated using GRADEpro software. The number of events of urgent visits to the hospital and requiring hospitalization was reduced (RR: 0.73; 95% CI: 0.69, 0.78; P value <0.00001). The mortality rate because of cardiovascular events was decreased with sotagliflozin (RR: 0.73; 95% CI: 0.67, 0.80; P value <0.00001). Patients taking sotagliflozin had a drastic decline in the number of deaths due to stroke and non-fatal myocardial infarction. Yet, there is no difference between the groups in terms of changes in mortality due to other causes or the glomerular filtration rate (GFR). Sotagliflozin demonstrated effectiveness in reducing the mortality rate related to heart failure and cardiovascular events when the dose was increased from 200 mg to 400 mg. Despite this, evidence is still needed to prove the renal protective action.

The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis and maintain translational repression of maternal mRNA and oocyte quality.

The regulation of maternal mRNAs is essential for proper oogenesis, the production of viable gametes, and to avoid birth defects and infertility. Many oogenic RNA-binding proteins have been identified with roles in mRNA metabolism, some of which localize to dynamic ribonucleoprotein granules and others that appear dispersed. Here, we use a combination of in vitro condensation assays and the in vivo Caenorhabditis elegans oogenesis model to characterize the properties of the conserved KH-domain MEX-3 protein and to identify novel regulators of MEX-3 and three other translational regulators. We demonstrate that MEX-3 undergoes phase separation and appears to have intrinsic gel-like properties in vitro. We also identify novel roles for the chaperonin-containing tailless complex polypeptide 1 (CCT) chaperonin and actin in preventing ectopic RNA-binding protein condensates in maturing oocytes that appear to be independent of MEX-3 folding. The CCT chaperonin and actin also oppose the expansion of endoplasmic reticulum sheets that may promote ectopic condensation of RNA-binding proteins. These novel regulators of condensation are also required for the translational repression of maternal mRNA which is essential for oocyte quality and fertility. The identification of this regulatory network may also have implications for understanding the role of hMex3 phase transitions in cancer.

The CCT chaperonin and actin modulate the ER and RNA-binding protein condensation during oogenesis to maintain translational repression of maternal mRNA and oocyte quality.

The regulation of maternal mRNAs is essential for proper oogenesis, the production of viable gametes, and to avoid birth defects and infertility. Many oogenic RNA-binding proteins have been identified with roles in mRNA metabolism, some of which localize to dynamic ribonucleoprotein granules and others that appear dispersed. Here, we use a combination of in vitro condensation assays and the in vivo C. elegans oogenesis model to determine the intrinsic properties of the conserved KH-domain MEX-3 protein and to identify novel regulators of MEX-3 and the Lsm protein, CAR-1. We demonstrate that MEX-3 undergoes liquid-liquid phase separation and appears to have intrinsic gel-like properties in vitro . We also identify novel roles for the CCT chaperonin and actin in preventing ectopic RNA-binding protein condensates in maturing oocytes that appear to be independent of MEX-3 folding. CCT and actin also oppose the expansion of ER sheets that may promote ectopic condensation of RNA-binding proteins that are associated with de-repression of maternal mRNA. This regulatory network is essential to preserve oocyte quality, prevent infertility, and may have implications for understanding the role of hMex3 phase transitions in cancer. Significance statement: The molecular mechanisms that regulate phase transitions of oogenic RNA-binding proteins are critical to elucidate but are not fully understood.We identify novel regulators of RNA-binding protein phase transitions in maturing oocytes that are required to maintain translational repression of maternal mRNAs and oocyte quality.This study is the first to elucidate a regulatory network involving the CCT chaperonin, actin, and the ER for phase transitions of RNA-binding proteins during oogenesis. Our findings for the conserved MEX-3 protein may also be applicable to better understanding the role of hMex3 phase transitions in cancer.

Synthesis and Structure Refinement of [Co-Al4-X] LDHs (X = NO3 - and SO4 2-) from Nordstrandite.

[M-Al4-X] LDHs (M = divalent metal and X = anion) are a class of aluminum-rich layered double hydroxides synthesized from both the gibbsite and bayerite polymorphs of Al(OH)3. Henceforth, "g" and "b" are used to indicate gibbsite and bayerite. Despite the differences in the stacking arrangement of the hydroxyl layers in the precursor polymorphs, [M-Al4-X] LDHs whether synthesized from gibbsite or bayerite were seen to be structurally the same. In this work, we report the first ever synthesis of [M-Al4-X] LDHs (M = Zn, Ni, and Co and X = NO3 - and SO4 2-) from nordstrandite, which is yet another polymorph of Al(OH)3. Hereafter, "n" represents the nordstrandite precursor. We report that n-[M-Al4-X] LDHs do not differ structurally from those prepared from gibbsite and bayerite. We also report the structural refinement of n-[Co-Al4-X] LDHs, where X = NO3 - and SO4 2-. This work is also significant as it gives for the very first time the refined structure of a [Co-Al4-NO3] LDH, though there are earlier reports on the synthesis of this LDH from both gibbsite and bayerite. The NO3 - ion in the interlayer makes an angle of ∼48° with the plane of the metal hydroxide layer, and its symmetry reduces from D 3h to C 2v . Similarly, the change in the symmetry of the SO4 2- ion in the interlayer is from T d to C 3v .

Systemic Review and Meta-Analysis to Evaluate Therapeutic Effectiveness of Interferon Beta-1b in Hospitalized COVID-19 Patients.

The COVID-19 pandemic has caused havoc in the health sector. Inflammatory cytokines play an important role in the disease condition. Existing evidence has provided certain insights into the repurposing of the drugs. This meta-analysis and systematic review aimed to explore the efficacy of the administration of interferon beta-1b (IFN ß-1b) and standard care versus only standard care as the therapeutic agent for managing COVID-19 patients who are severely ill. The search was conducted in the following databases: Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, Scopus, and Google Scholar, which were published during the period January 1, 2020, to February 16, 2023. All the included three studies were independently assessed for eligibility. The modified data extraction form of Cochrane were used. The quality of the three included studies was assessed using the Cochrane risk of bias tool. GradePro software was used to summarize the quality grading of the primary outcome measures. The time taken for clinical improvement was (MD: -3.28 days; 95% CI: -5.65, -0.91; P value = 0.007) when treated with IFN ß-1b. The duration of hospital stays (MD: -2.43 days; 95% CI: -4.45, -0.30; P value = 0.03), and need for intensive care unit (ICU) admission (RR: 0.71; 95% CI: 0.52, 0.97; P value = 0.03) was statistically significant. Interferon beta-1b is proven to reduce the duration of hospital stay, and the improved clinical status may become a cornerstone of COVID-19 treatment.

Dehydration-Rehydration Studies on Polytypes of Chloride and Nitrate Layered Double Hydroxides of Nordstrandite and Bayerite: a Comparative Study.

In this work, we report for the first time, the dehydration-rehydration studies of nordstrandite-derived layered double hydroxides (LDHs) of Li and Al, n-[Li-Al-X] (X = Cl- and NO3 -) (n-nordstrandite derived). n-[Li-Al-NO3], an orthorhombic phase, dehydrated at 180 °C to a monoclinic phase. Refinement placed the NO3 - ions parallel to the hydroxide layers. The dehydration showed no change in basal spacing. The monoclinic n-[Li-Al-Cl] dehydrated at 160 °C with a 0.49 Å compression in basal spacing to an orthorhombic polytype. We compared our results with the published results of their bayerite counterparts b-[Li-Al-X] (b-bayerite derived) and observed that though n-[Li-Al-X] and b-[Li-Al-X] LDHs have similar structures, their dehydrated phases are structurally different. We also report the refinement of b-[Li-Al-Cl] (DH). Previous studies attribute the basal spacing values to the (i) degree of hydration and (ii) orientation of anions in the interlayer. We observe that basal spacing is a manifestation of the symmetry of the crystal. Dehydration of nitrate intercalated LDH, which proceeds from an orthorhombic symmetry to a monoclinic symmetry with no decrease in the interlayer spacing, is attributed to sliding of the hydroxyl layers in the ab-plane due to the increase in the ß value. This sliding stabilizes the interlayer through weak long-range electrostatic forces that mainly contribute to the stabilization of the layered structure at separations much larger than the effective radius of hydrogen bonds. Such stabilization would negate the need for the layers to compress, thus conserving the basal spacing in n-[Li-Al-NO3] (DH).

Dosage Optimization of Lamotrigine in Pregnancy: A Pharmacometric Approach Using Modeling and Simulation.

Lamotrigine is the most widely used anti-epileptic drug in pregnancy because of its low teratogenicity. However, there is an increased metabolism and clearance of lamotrigine in pregnancy contributing to suboptimal drug therapy and poor disease control, prompting the need for proactive dosage adjustments. The present study aimed to develop a pharmacometric model-based framework for recommending an optimal dosage regimen for lamotrigine in pregnancy. A systematic review was performed to obtain aggregate data from the literature on the clearance of lamotrigine in pregnancy. The data were incorporated into simulations using PUMAS software to estimate the plasma concentrations at the preconception stage and during the 3 trimesters of pregnancy. Simulated drug exposures for different doses were investigated to ascertain plasma concentrations similar to the preconception value and above the minimum effective concentration. The simulated mean steady-state trough plasma concentrations of lamotrigine were significantly decreased in pregnant women over the course of the 3 trimesters (3.17 ± 0.93 mg/L, 2.14 ± 0.86 mg/L, and 1.51 ± 0.65 mg/L, respectively), compared with non-pregnant women (4.31 ± 1.14 mg/L) (P < .001). The simulation studies revealed that doses of 150 mg, 175 mg, 225 mg, and 250 mg twice daily in the preconception stage and the 3 trimesters, respectively, achieve the target concentrations. Thus, the model-informed dosage regimen of lamotrigine proposed in this study shall be used to initiate appropriate dosing in pregnant women; however, the safety and efficacy of the drug must be assured through therapeutic drug monitoring in order to avoid therapeutic failure of lamotrigine in pregnancy.

Strengthening global midwifery education to improve quality maternity care: Co-designing the World Health Organization Midwifery Assessment Tool for Education (MATE).

AIMS: To describe a three-phase co-designed project to develop a culturally appropriate and relevant education assessment tool, and report on pilot and field-testing phases. BACKGROUND: High-quality midwifery education is essential for high-quality maternity care (WHO 2019); however midwifery education and maternity care vary in quality throughout Europe. To support countries in strengthening their midwifery education, World Health Organization (WHO) European Region commissioned development of the Midwifery Assessment Tool for Education (MATE). The tool was developed over three years, using an iterative, collaborative process with regional experts. Published by WHO in May 2020, MATE provides focused questions and evidence-informed resources to stimulate and inform discussions within country. DESIGN: Three-phase co-design approach to develop, pilot and field-test an education assessment tool. METHODS: Phase 1: initial development of MATE with expert midwifery support; Phase 2: MATE piloting workshops in Czech Republic and Lithuania focusing on clarity, usability and relevance; Phase 3: MATE field-testing workshop in Bulgaria exploring the process of using MATE and its effectiveness for generating discussion. Purposive selection of workshop participants ensured a broad range of perspectives: clinicians, educators, students, policy makers and service users. All participants were invited to give narrative feedback during workshops and via completion of a post-workshop online survey. The XX University Research Ethics Committee advised that formal ethical review was unnecessary. RESULTS: Feedback from collaborators in all phases indicated that engaging with MATE co-design and testing was a positive experience. A 'bottoms up' approach ensured that MATE content was relevant to regional needs, culturally acceptable and appropriate. Seventy-nine individuals participated in Phases 2 and 3 and all were sent a post-workshop online survey, with 31 responses (39 %). Qualitative and quantitative data indicated that the aim of MATE was well understood, and its usability and relevance were evaluated positively. In Phase 2, improvements to wording and format were suggested. MATE was subsequently amended prior to field testing. Phase 3 feedback indicated that MATE was highly effective for generating in-country dialogue and frank discussions about the future of midwifery education and practice. CONCLUSIONS: Using a co-design approach has ensured that MATE is culturally relevant, accessible and appropriate. This initial evaluation indicates that MATE can facilitate in-country dialogue and support the strengthening of midwifery education in accordance with WHO aims. Next steps are a fully evaluated trial of MATE in a selected partner country, where we will continue to work collaboratively to optimise engagement and ensure cultural appropriateness.

Anti-cardiolipin IgG autoantibodies associate with circulating extracellular DNA in severe COVID-19.

Whereas the detection of antiphospholipid autoantibodies (aPL) in COVID-19 is of increasing interest, their role is still unclear. We analyzed a large aPL panel in 157 patients with COVID-19 according to the disease severity. We also investigated a potential association between aPL and extracellular DNA (exDNA, n = 85) or circulating markers of neutrophil extracellular traps (NET) such as citrullinated histones H3 (CitH3, n = 49). A total of 157 sera of patients infected by SARS-CoV-2 were collected. A large aPL panel including lupus anticoagulant, anti-cardiolipin and anti-beta-2 glycoprotein I (IgG, IgM and IgA), anti-phosphatidylethanolamine IgA, anti-prothrombin (IgG and IgM) was retrospectively analyzed according to the disease severity. We found a total aPL prevalence of 54.8% with almost half of the cases having aCL IgG. Within an extended panel of aPL, only aCL IgG were associated with COVID-19 severity. Additionally, severe patients displayed higher CitH3 levels than mild patients. Interestingly, we highlighted a significant association between the levels of aCL IgG and exDNA only in aCL positive patients with severe disease. In conclusion, we showed a significant link between aPL, namely aCL IgG, and circulating exDNA in patients with severe form of COVID-19, that could exacerbate the thrombo-inflammatory state related to disease severity.

A single chiroptical spectroscopic method may not be able to establish the absolute configurations of diastereomers: dimethylesters of hibiscus and garcinia acids.

Electronic circular dichroism (ECD), optical rotatory dispersion (ORD), and vibrational circular dichroism (VCD) spectra of hibiscus acid dimethyl ester have been measured and analyzed in combination with quantum chemical calculations of corresponding spectra. These results, along with those reported previously for garcinia acid dimethyl ester, reveal that none of these three (ECD, ORD, or VCD) spectroscopic methods, in isolation, can unequivocally establish the absolute configurations of diastereomers. This deficiency is eliminated when a combined spectral analysis of either ECD and VCD or ORD and VCD methods is used. It is also found that the ambiguities in the assignment of absolute configurations of diastereomers may also be overcome when unpolarized vibrational absorption is included in the spectral analysis.

Forced association of SARS-CoV-2 proteins with the yeast proteome perturb vesicle trafficking.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the highly infectious coronavirus disease COVID-19. Extensive research has been performed in recent months to better understand how SARS-CoV-2 infects and manipulates its host to identify potential drug targets and support patient recovery from COVID-19. However, the function of many SARS-CoV-2 proteins remains uncharacterised. Here we used the Synthetic Physical Interactions (SPI) method to recruit SARS-CoV-2 proteins to most of the budding yeast proteome to identify conserved pathways which are affected by SARS-CoV-2 proteins. The set of yeast proteins that result in growth defects when associated with the viral proteins have homologous functions that overlap those identified in studies performed in mammalian cells. Specifically, we were able to show that recruiting the SARS-CoV-2 NSP1 protein to HOPS, a vesicle-docking complex, is sufficient to perturb membrane trafficking in yeast consistent with the hijacking of the endoplasmic-reticulum-Golgi intermediate compartment trafficking pathway during viral infection of mammalian cells. These data demonstrate that the yeast SPI method is a rapid way to identify potential functions of ectopic viral proteins.