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Compact cell type-specific promoters are important tools for basic and preclinical research and clinical delivery of gene therapy. In this work, we designed novel MiniPromoters to target D1 and D2 type dopaminoceptive medium spiny neurons in the striatum by manually identifying candidate regulatory regions or employing the OnTarget webserver. We then empirically tested the designs in rAAV-PHP.B for specificity and robustness in three systems: intravenous injection in mice, intracerebroventricular injection in mice, and intracerebroventricular injection in non-human primates. Twelve MiniPromoters were designed from eight genes: seven manually and five using OnTarget. When delivered intravenously in mice, three MiniPromoters demonstrated highly selective expression in the striatum, with Ple389 (ADORA2A) showing high levels of dopamine D2-receptor cell co-localization. The same three MiniPromoters also displayed enriched expression in the striatum when delivered intracerebroventricularly in mice with high levels of DARPP32 co-localization. Finally, Ple389 (ADORA2A) was intracerebroventricularly injected in non-human primates and showed enriched expression in the striatum as in the mouse. Ple389 (ADORA2A) demonstrated expression in the medium spiny neurons in all three systems tested and exhibited the highest level of D2-MSNs and DARPP32 co-labeling in mice, demonstrating its potential as a tool for gene therapy approaches for Parkinson and Huntington disease treatment., Competing Interests: Declarations Competing interests The authors declare no competing interests., (© 2024. The Author(s).)

Arylalkylamine N-acetyltransferase (AANAT) catalyzes the rate-limiting step in melatonin synthesis and is a potential target for disorders involving melatonin overproduction, such as seasonal affective disorder. Previously described AANAT inhibitor bromoacetyltryptamine (BAT) and benzothiophenes analogs were reported to react with CoASH to form potent bisubstrate inhibitors through AANAT's alkyltransferase function, which is secondary to its role as an acetyltransferase. We replaced the bromoacetyl group in BAT with various Michael acceptors to mitigate possible off-target activity of its bromoacetyl group. Additionally, we modified the length of the carbon linker between the Michael acceptor and indole bicycle of tryptamine to determine its effect on potency. An AANAT enzymatic assay showed a two-carbon linker present in BAT was optimal, while none of the new warheads had activity. Kinetic analysis indicated that these Michael acceptors reacted with CoASH much slower than BAT and not within the timeframe of our enzymatic assay. Additionally, we confirmed earlier reports that the acetyltransferase function of AANAT follows an ordered bi bi mechanism in which AcCoA binds before serotonin. In contrast, BAT's alkyltransferase kinetics revealed a bi uni mechanism in which BAT binds to AANAT before CoASH. Our model combines both functions of AANAT into one kinetic mechanism., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction: The English National Health Service (NHS) is the first to offer whole genome sequencing (WGS) as part of standard care. As a high-income country with a universal healthcare system, England contributes a valuable perspective to global developments in WGS., Methods: We used an implementation science approach with mixed methods to characterise delivery of WGS for paediatric rare diseases. Observations and field notes of consent appointments in clinical genetics and mainstream settings and follow-up qualitative semi-structured interviews with the clinical team. Process maps were developed for each department to identify similarities and variations between sites, and thematic analysis of interview data to understand barriers and facilitators., Results: Data collection occurred in 12 departments (7 genetic, 3 neurology, 1 cardiology and 1 general paediatric) across 7 NHS Trusts. 26 observations of 21 healthcare professionals were conducted, alongside 19 follow-up interviews. Two master maps were developed - one for clinical genetics and one for the mainstream. We identified 11 steps involved in delivering WGS, including 9 variations and 9 similarities. We identified most variation in the processes related to the 'who', 'when', 'how' and 'where' as these were aspects that could be adapted to fit into the specific set-up of the department. Barriers included reluctance to uptake in the mainstream and difficulties tracking samples., Conclusion: Recommendations include developing SOPs and hiring healthcare professionals responsible for facilitating consent alongside administrative aspects. These would reduce the burden on clinical geneticists and improve turnaround times as well as contribute to streamlining and standardisation of the service., (The Author(s). Published by S. Karger AG, Basel.)

Oligodendrocytes (OLs) of the central nervous system require iron for proteolipid biosynthesis during the myelination process. Although most heme is found complexed to hemoglobin in red blood cells, surprisingly, we found that Slc48a1, encoding the heme transporter Hrg1, is expressed at higher levels in OLs than any other cell type in rodent and humans. We confirmed in situ that Hrg1 is expressed in OLs but not their precursors (OPCs) and found that Hrg1 proteins in CNS white matter co-localized within myelin sheaths. In older Hrg1 null mutant mice we observed reduced expression of myelin associated glycoprotein (Mag) and ultrastructural myelin defects reminiscent of Mag-null animals, suggesting myelin adhesion deficiency. Further, we confirmed reduced myelin iron levels in Hrg1 null animals in vivo, and show that OLs in vitro can directly import both the fluorescent heme analogue ZnMP and heme itself, which rescued iron deficiency induced inhibition of OL differentiation in a heme-oxidase-dependent manner. Together these findings indicate OL Hrg1 encodes a functional heme transporter required for myelin integrity., (© 2024 The Author(s). GLIA published by Wiley Periodicals LLC.)

Participants in the 100,000 Genomes Project (100kGP) could consent to receive additional finding (AF) results, individual variants relating to genes associated with susceptibility to cancer and familial hypercholesterolemia (FH). In the study reported here, qualitative interviews were used to explore the experiences of National Health Service (NHS) professionals from across England who were tasked with returning over 80,000 "no AF" results and 700 positive AF results to 100kGP participants. Interviews were conducted with 45 professionals from a range of backgrounds, including Genetic Counsellors, Clinical Geneticists, FH Clinical Nurse Specialists and Clinical Scientists. Interviews were analysed using a codebook thematic analysis approach. Returning AF results has been a significant endeavour, with challenges for pathways, administrative processes and clinical and laboratory time when the capacity of NHS services is already stretched. Professionals discussed going "above and beyond" to prioritise patient care through pathway design, additional clinics, overtime, longer appointments and provision of follow-up appointments. Professionals also described facing practical and emotional challenges when returning AFs. Benefits for patients from receiving AFs in the 100kGP were highlighted and professionals were generally positive about offering clinically actionable AFs within routine NHS clinical care. Professionals were, however, cautious around the implementation of AFs into routine care and felt more research and discussion was needed to determine which AFs to offer, approaches to consent and communication of results, costs and the potential strain on NHS capacity and resources. Further consultation is required with careful review of pathways and resources before offering AFs in clinical practice., (© 2024. The Author(s).)

Background: While the estimated number of U.S. influenza-associated deaths is reported annually, detailed data on the epidemiology of influenza-associated deaths, including the burden of in-hospital versus post-hospital discharge deaths are limited., Methods: Using data from the 2010-11 through 2018-19 seasons from the Influenza Hospitalization Surveillance Network, we linked cases to death certificates to identify patients who died from any cause during their influenza hospital stay or within 30 days post discharge. We described demographic and clinical characteristics of patients who died in hospital versus post discharge and characterized locations and causes of death (COD)., Results: Among 121,390 cases hospitalized with laboratory-confirmed influenza over 9 seasons, 5.5% died; 76% of deaths were in patients ≥65 years, 71% were non-Hispanic White, and 34% had ≥4 underlying medical conditions. Among all patients with an influenza-associated hospitalization who died, 48% of deaths occurred after hospital discharge; the median days from discharge to death was 9 days (IQR 3-19 days). Post-discharge deaths more often occurred in older patients and among those with underlying medical conditions. Only 37% of patients who died had "influenza" as a COD on their death certificate. Influenza was more frequently listed as a COD among persons who died in-hospital compared with cardiovascular disease among those who died after discharge., Conclusions: All-cause mortality burden is substantial among patients hospitalized with influenza, with almost 50% of deaths occurring within 30 days after hospital discharge. Surveillance systems should consider capture of post-discharge outcomes to better characterize the impact of influenza on all-cause mortality., (Published by Oxford University Press on behalf of Infectious Diseases Society of America 2024.)

Caffeine consumption can elicit improvements in aspects of physical function in older adults but also, negatively modify standing balance, potentially increasing fall risk. However, balance alterations and changes in physical function induced by commonly consumed caffeine vehicles such as coffee have not been investigated. Therefore, this study investigated coffee ingestion providing 3 mg·kg BW -1 caffeine on balance performance and physical function, in a group of older adults. In a randomised, crossover design, 22 older adults (Male n = 10, Age: 68 ± 6 years) completed bipedal standing balance and physical function assessments (Senior Fitness Test) under one of the following conditions: caffeinated coffee (COF), decaffeinated coffee (DEC), placebo (PLA) or a control (CON) (no fluid ingestion). Centre of pressure (COP) root mean square and power frequency were calculated to characterise postural performance and strategy, respectively. The complexity (i.e., regularity) of the COP signal was also determined by calculating sample entropy. Caffeinated coffee had limited effects on COP outcomes. Frequency of the COP in the anteroposterior direction was greater following COF compared to DEC (P = 0.047;g = 0.29) but there were no statistical differences between COF and PLA or CON (P > 0.05). Furthermore, there were no significant performance differences between any conditions in all tests of physical function (P > 0.05). This suggests that coffee has limited effects on balance performance or physical function but may influence both balance complexity and the strategy utilised to maintain upright stance. Overall, a strong cup of coffee does not significantly influence balance and measures of functional performance in healthy older adults., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Survivors of childhood sexual abuse (CSA) often experience long-term adverse mental health effects, a trend that has been observed in research focusing on men who have sex with men (MSM), especially Black MSM., Objective: The aim of this study was to investigate the direct and indirect effects of childhood sexual abuse on depression symptoms among Black MSM through early sexual debut, histories of incarceration, ethnic identity, and social support. In addition, we examine the role of social support and ethnic identity as mediators of depression symptoms., Participants and Setting: The HPTN 073 study enrolled and followed 226 HIV-uninfected Black MSM in three US cities (Los Angeles; Washington, DC; and Chapel Hill, North Carolina) from February 2013 to September 2015. Study participants were offered once-daily oral emtricitabine/tenofovir preexposure prophylaxis combined with counseling and followed for 52 weeks., Methods: A path analysis was used to examine direct and indirect effects of CSA experiences on depression symptoms through incarceration, early sexual debut ethnic identity, and social support, and to see whether social support and ethnic identity mediated the relationship between incarceration and depression symptoms., Results: Our results indicate that childhood sexual abuse was direct and positively associated with early sexual debut (β = 0.21, p < .001). Both ethnic identity (β = -0.14, p < .001) and social support (β = -0.82, p < .001) were direct and negatively associated with depressive symptoms., Conclusion: Our research underscores the significant impact of CSA factors on the life trajectories of some Black MSM, including experiences such as incarceration, sexual debut, and depression symptoms., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The immunoregulatory cation channel TMEM176B plays a dual role in tumor immunity. On the one hand, TMEM176B promotes antigen cross-presentation to CD8 + T cells by regulating phagosomal pH in dendritic cells (DCs). On the other hand, it inhibits NLRP3 inflammasome activation through ionic mechanisms in DCs, monocytes and macrophages. We speculated that formulating BayK8644 in PEGylated chitosan nanoparticles (NP-PEG-BayK8644) should slowly release the compound and by that mean avoid cross-presentation inhibition (which happens with a fast 30 min kinetics) while still triggering inflammasome activation. Chitosan nanocarriers were successfully obtained, exhibiting a particle size within the range of 200 nm; they had a high positive surface charge and a 99 % encapsulation efficiency. In in vitro studies, NP-PEG-BayK8644 did not inhibit antigen cross-presentation by DCs, unlike the free compound. The NP-PEG-BayK8644 activated the inflammasome in a Tmem176b-dependent manner in DCs. We administered either empty (eNP-PEG) or NP-PEG-BayK8644 to mice with established tumors. NP-PEG-BayK8644 significantly controlled tumor growth and improved mice survival compared to both eNP-PEG and free BayK8644 in melanoma and lymphoma models. This effect was associated with enhanced inflammasome activation by DCs in the tumor-draining lymph node and infiltration of the tumor by CD8 + T cells. Thus, encapsulation of BayK8644 in chitosan NPs improves the anti-tumoral properties of the compound by avoiding inhibition of antigen cross-presentation., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: MH is founder and CSO of ARDAN Pharma. One patent application related to this work has been filed at the USA Patent and Trademark Office., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Following ICH guidelines for analytical validation, we report a common C18 column stability indicating isocratic reverse phase high performance liquid chromatography method for the determination of the ion channel modulator Bay K8644. Two main forced degradation products and a minor impurity were also tentatively identified by Mass Spectrometry. The mobile phase consisted of a 50/50 acetonitrile/buffer mixture at a flow rate of 2 mL/min. Mean retention time for Bay K8644 was 3.030 minutes. Excellent linearity (r = 0.9998) was achieved in the range 0.10-1.40 μg/mL at 274 nm wavelength. Analytical limits were 16.56 ± 1.04 ng/mL for detection and 55.21 ± 3.48 ng/mL for quantitation respectively. Accuracy and precision studies showed good results (95-105%). Robustness was assessed by varying ±3%, both temperature and flow rate. Five different stress conditions were applied to assess Bay K8644's stability. Only basic and photolytic treatments yielded degradation products, both correctly resolved in a total runtime of 4 minutes. In conclusion, we developed a fast, simple, sensitive, accurate, precise, reliable and stability indicating method for detecting/quantifying Bay K8644, and tentatively characterized its main impurities/forced degradation products., (© The Author(s) 2024. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Acquired generalized lipodystrophy (AGL) is a rare disease characterized by variable loss of adipose tissue and concurrent metabolic derangements, typically with childhood or adolescent onset. AGL has three subclassifications: panniculitis (type 1), autoimmune disease (type 2), and idiopathic (type 3). This report highlights a rare case of AGL type 1 in a previously healthy 3-year-old female who presented with diffuse erythematous subcutaneous nodules, progressive lipoatrophy, and histopathological findings of a lobular panniculitis., (© 2024 Wiley Periodicals LLC.)

Pyometra is rare and refers to purulent material in the uterine cavity, which can lead to uterine perforation. A hysterectomy is the historic treatment for perforated pyometra due to the condition's high morbidity and mortality. A 33-year-old woman with uterine factor infertility underwent an abdominal myomectomy that was complicated by post-operative, perforated pyometra. She received conservative treatment with surgical debridement and broad-spectrum antibiotics. Following her recovery and subsequent infertility treatments, she had a term live birth through in vitro fertilization. The case highlights the value of uterine-sparing treatment in carefully selected patients with post-operative, perforated pyometra, in order to preserve fertility., (© 2024 Published by Elsevier B.V.)

Background: Care partners are individuals chosen by a person with an illness to support their care during hospitalization. Patients with persistent critical illness have longer than average critical care admission and often other conditions including dysphagia, communication vulnerability, severe physical deconditioning, the need for an artificial airway, and difficulty weaning from invasive mechanical ventilation. Family presence has been identified as important for patients experiencing persistent critical illness in specialized weaning centers. Despite this, the role of care partners in clinical settings for patients with persistent critical illness has not been fully characterized, particularly from the perspectives of patients, care partners, and health care providers. The aim of this study was to gain insights into the roles of care partners during persistent critical illness from the perspectives of patients, care partners, and health care providers., Methods: We used qualitative descriptive methodology including semi-structured interviews and content analysis. Interviews were audio recorded and transcribed verbatim. Included participants ( n  = 30) were patient survivors ( n  = 7), care partners of patient survivors ( n  = 9), and professionally diverse health care providers ( n  = 14) of adult patients with persistent critical illness from two specialized units in one community academic hospital in Toronto, Canada., Results: Participants across all groups described care partner roles that included physical, mental health, cognitive, social, and spiritual support of the patient, including the perceived role of safeguarding the multiple dimensions of care for the patient who is experiencing persistent critical care in specialized care settings., Discussion: The results of this study are being used to co-design, implement, and evaluate a sustainable care partner program that is acceptable, appropriate, and feasible to implement in clinical settings where the care of patients with persistent critical illness occurs., Reporting Method: Consolidated criteria for reporting qualitative studies (COREQ)., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Istanboulian, Gilding, Hamilton, Master, Bingler, Hill, Isani, kazi, Coppinger and Smith.)

Diabetes is a major risk factor for Alzheimer's disease (AD). Amino acid compound 2 (AAC2) improves glycemic and cognitive functions in diabetic mouse models through mechanisms distinct from insulin. Our goal was to compare the effects of AAC2, insulin, and their nanofiber-forming combination on early asymptomatic AD pathogenesis in APP/PS1 mice. Insulin, but not AAC2 or the combination treatment (administered intraperitoneally every 48 h for 120 days), increased seizure-related mortality, altered the brain fat-to-lean mass ratio, and improved specific cognitive functions in APP/PS1 mice. NanoString and pathway analysis of cerebral gene expression revealed dysregulated synaptic mechanisms, with upregulation of Bdnf and downregulation of Slc1a6 in insulin-treated mice, correlating with insulin-induced seizures. In contrast, AAC2 promoted the expression of Syn2 and Syp synaptic genes, preserved brain composition, and improved survival. The combination of AAC2 and insulin counteracted free insulin's effects. None of the treatments influenced canonical amyloidogenic pathways. This study highlights AAC2's potential in regulating synaptic gene expression in AD and insulin-induced contexts related to seizure activity.

Mutations in human VPS4A are associated with neurodevelopmental defects, including motor delays and defective muscle tone. VPS4A encodes a AAA-ATPase required for membrane scission, but how mutations in VPS4A lead to impaired control of motor function is not known. Here we identified a mutation in zebrafish vps4a , T248I, that affects sensorimotor transformation. Biochemical analyses indicate that the T248I mutation reduces the ATPase activity of Vps4a and disassembly of ESCRT filaments, which mediate membrane scission. Consistent with the role for Vps4a in exosome biogenesis, vps4a T248I larvae have enlarged endosomal compartments in the CNS and decreased numbers of circulating exosomes in brain ventricles. Resembling the central form of hypotonia in VPS4A patients, motor neurons and muscle cells are functional in mutant zebrafish. Both somatosensory and vestibular inputs robustly evoke tail and eye movements, respectively. In contrast, optomotor responses, vestibulospinal, and acoustic startle reflexes are absent or strongly impaired in vps4a T248I larvae, indicating a greater sensitivity of these circuits to the T248I mutation. ERG recordings revealed intensity-dependent deficits in the retina, and in vivo calcium imaging of the auditory pathway identified a moderate reduction in afferent neuron activity, partially accounting for the severe motor impairments in mutant larvae. Further investigation of central pathways in vps4a T248I mutants showed that activation of descending vestibulospinal and midbrain motor command neurons by sensory cues is strongly reduced. Our results suggest that defects in sensorimotor transformation underly the profound yet selective effects on motor reflexes resulting from the loss of membrane scission mediated by Vps4a. Significance Statement Here we present a T248I mutation in vps4a , which causes sensorimotor defects in zebrafish larvae. Vps4a plays a key role in membrane scission. Spanning biochemical to systems level analyses, our study indicates that a reduction in Vps4a enzymatic activity leads to abnormalities in membrane-scission dependent processes such as endosomal protein trafficking and exosome biogenesis, resulting in pronounced deficits in sensorimotor transformation of visual, auditory, and vestibular cues. We suggest that the mechanisms underlying this type of dysfunction in zebrafish may also contribute to the condition seen in human patients with de novo mutations in the human VPS4A orthologue., (Copyright © 2024 the authors.)

Background: In the EMPA-KIDNEY trial, empagliflozin, a sodium-glucose cotransporter 2 (SGLT2) inhibitor, had positive cardiorenal effects in patients with chronic kidney disease who were at risk for disease progression. Post-trial follow-up was designed to assess how the effects of empagliflozin would evolve after the discontinuation of the trial drug., Methods: In the active trial, patients with chronic kidney disease were randomly assigned to receive either empagliflozin (10 mg once daily) or matching placebo and were followed for a median of 2 years. All the patients had an estimated glomerular filtration rate (eGFR) of at least 20 but less than 45 ml per minute per 1.73 m 2 of body-surface area or an eGFR of at least 45 but less than 90 ml per minute per 1.73 m 2 with a urinary albumin-to-creatinine ratio (with albumin measured in milligrams and creatinine measured in grams) of at least 200. Subsequently, surviving patients who consented were observed for 2 additional years. No trial empagliflozin or placebo was administered during the post-trial period, but local practitioners could prescribe open-label SGLT2 inhibitors, including open-label empagliflozin. The primary composite outcome was kidney disease progression or cardiovascular death as assessed from the start of the active-trial period to the end of the post-trial period., Results: Of the 6609 patients who had undergone randomization in the active trial, 4891 (74%) were enrolled in the post-trial period. During this period, the use of open-label SGLT2 inhibitors was similar in the two groups (43% in the empagliflozin group and 40% in the placebo group). During the combined active- and post-trial periods, a primary-outcome event occurred in 865 of 3304 patients (26.2%) in the empagliflozin group and in 1001 of 3305 patients (30.3%) in the placebo group (hazard ratio, 0.79; 95% confidence interval [CI], 0.72 to 0.87). During the post-trial period only, the hazard ratio for a primary-outcome event was 0.87 (95% CI, 0.76 to 0.99). During the combined periods, the risk of kidney disease progression was 23.5% in the empagliflozin group and 27.1% in the placebo group; the risk of the composite of death or end-stage kidney disease was 16.9% and 19.6%, respectively; and the risk of cardiovascular death was 3.8% and 4.9%, respectively. There was no effect of empagliflozin on death from noncardiovascular causes (5.3% in both groups)., Conclusions: In a broad range of patients with chronic kidney disease at risk for progression, empagliflozin continued to have additional cardiorenal benefits for up to 12 months after it was discontinued. (Funded by Boehringer Ingelheim and others; EMPA-KIDNEY ClinicalTrials.gov number, NCT03594110; EuDRACT number, 2017-002971-24.)., (Copyright © 2024 Massachusetts Medical Society.)

Adiposity is an established risk factor for multiple diseases, but the causal relationships of different adiposity types with circulating protein biomarkers have not been systematically investigated. We examine the causal associations of general and central adiposity with 2923 plasma proteins among 3977 Chinese adults (mean BMI = 23.9 kg/m²). Genetically-predicted body mass index (BMI), body fat percentage (BF%), waist circumference (WC), and waist-to-hip ratio (WHR) are significantly (FDR < 0.05) associated with 399, 239, 436, and 283 proteins, respectively, with 80 proteins associated with all four and 275 with only one adiposity trait. WHR is associated with the most proteins (n = 90) after adjusting for other adiposity traits. These associations are largely replicated in Europeans (mean BMI = 27.4 kg/m²). Two-sample Mendelian randomisation (MR) analyses in East Asians using cis-protein quantitative trait locus (cis-pQTLs) identified in GWAS find 30/2 proteins significantly affect levels of BMI/WC, respectively, with 10 showing evidence of colocalisation, and seven (inter-alpha-trypsin inhibitor heavy chain H3, complement factor B, EGF-containing fibulin-like extracellular matrix protein 1, thioredoxin domain-containing protein 15, alpha-2-antiplasmin, fibronectin, mimecan) are replicated in separate MR using different cis-pQTLs identified in Europeans. These findings identified potential novel mechanisms and targets, to our knowledge, for improved treatment and prevention of obesity and associated diseases., (© 2024. The Author(s).)

Purpose: The modified Rankin Scale (mRS) at 90 days is the primary outcome in most acute stroke studies, but the long follow-up period has disadvantages. The National Institutes of Health Stroke Scale (NIHSS) at 24 h shows a strong, but imperfect, association with 90-day mRS. This study examines the association between 24-hour NIHSS and 90-day mRS and reasons for discrepancies., Methods: Data are from the ESCAPE-NA1 thrombectomy patients. To address the non-normality distribution of the NIHSS and include deceased patients, a 7-point ordinal score was generated by grouping 24-hour NIHSS. The association of ordinal 24-hour NIHSS and 90-day mRS was assessed with adjusted ordinal logistic regression. Differences in baseline and treatment/post-treatment variables were compared between patients with discordant and concordant outcomes., Results: One-thousand-seventy-six patients with available 24-hour NIHSS and 90-day mRS were included (median 24-hour NIHSS 6[IQR: 2-14], median 90-day mRS 2[IQR: 1-4]). Ordinal 24-hour NIHSS was associated with 90-day mRS (adjusted cOR 2.53 [95%CI 2.33-2.74]). Forty-eight (4.5%) patients had discordant outcomes. Of those, 19(1.8%) had 24-hour NIHSS < 6 and 90-day mRS5-6; all of which had ≥ 1 severe adverse event, most commonly pneumonia (6[31.6%]) or recurrent stroke (4[21.1%]). Twenty-nine patients (2.7%) had 24-hour NIHSS > 14 and 90-day mRS 0-2. In these patients, baseline NIHSS and ASPECTS was lower, and collateral status was worse., Conclusion: An ordinal NIHSS score that includes death at 24 h shows a strong association with 90-day mRS, suggesting that it could be used as an alternative outcome. Patients with discrepant outcomes differed from the remaining patients regarding their baseline NIHSS, ASPECTS, collateral status, and post-stroke complications., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany.)

Objectives: To examine cardiovascular autonomic function in middle-aged people with long-term cervical and upper thoracic spinal cord injury (SCI) compared with the general population, and explore if the neurological level of injury (NLI) is related to cardiovascular autonomic function., Design: Population-based cross-sectional study with matched controls., Setting: Outpatient SCI unit in Southern Sweden., Participants: Twenty-five individuals (20% women, mean age 58 years and mean time since injury 28 years, NLI C2-T6, American Spinal Injury Association Impairment Scale A-C) from the Swedish SPinal Cord Injury Study on Cardiopulmonary and Autonomic Impairment (SPICA). Matched controls were obtained from the population-based Swedish CArdioPulmonary bioImage Study (SCAPIS) at a ratio of 5:1., Interventions: Not applicable., Outcome Measures: 24 h electrocardiography and deep breathing tests. 24 h ambulatory blood pressure (BP) monitoring and orthostatic BP tests., Results: In individuals with SCI compared with controls, heart rate variability (24h mean SD of the normal-to-normal interval 112 ms vs 145 ms, P < 0.001) and diastolic orthostatic BP increase (2.0 and 9.4 mmHg, P < 0.001), were significantly lower, whereas BP variability was significantly higher (24h mean systolic SD BP 17.8 mmHg vs 15.7 mmHg, P = 0.029). Circadian patterns of heart rate variability and BP (lack of nocturnal dip) were significantly different among the individuals with SCI than controls. Higher NLI was significantly (P < 0.05) correlated with impairments to various cardiovascular autonomic function variables., Conclusions: This exploratory study indicates that cardiovascular autonomic function is impaired in middle-aged people with long-term cervical and upper thoracic SCI compared with the general non-SCI population, and more pronounced with a higher NLI. Future research is needed to understand the pathophysiological mechanisms underlying these impairments, and the prognostic significance for individuals with SCI., Trial Registration: ClinicalTrials.gov identifier: NCT03515122.

Background: People living with HIV experience HIV stigma alongside a spectrum of aging-related health conditions that accelerate their vulnerability to the ill effects of loneliness and social isolation. Group-singing interventions are efficacious in improving psychosocial well-being among older people in the general population; however, the social curative effects of group singing have not been explored in relation to HIV stigma. By promoting group identification, bonding, and pride, group singing may reduce loneliness, social isolation, and other negative impacts of HIV stigma among older people living with HIV. Access to group-singing programs may be enhanced by technology., Objective: While group singing has been extensively studied in older adults, group-singing interventions have not been adapted for older people living with HIV to target loneliness and social isolation in the context of HIV stigma. The objective of this study was to describe the systematic development of a group-singing intervention to reduce loneliness and social isolation among older people living with HIV., Methods: In the San Francisco Bay Area between February 2019 and October 2019, we engaged older people living with HIV in a rigorous, 8-stage, community-engaged intervention adaptation process using the Assessment, Decision, Adaptation, Production, Topical Experts, Integration, Training, and Testing (ADAPT-ITT) framework. On the basis of a formative assessment of the needs and preferences of older people living with HIV, we selected an evidence-based group-singing intervention for older adults and systematically adapted the intervention components by administering them to a community advisory council (n=13)., Results: The result was United Voices, a 12-week hybrid (web-based and in-person) group-singing intervention for older people living with HIV. United Voices comprises 12 web-based (ie, via Zoom [Zoom Video Communications]) rehearsals, web-based and in-person drop-in helpdesk sessions, and a professionally produced final concert recording., Conclusions: Through an iterative process and in consultation with stakeholders and topic experts, we refined and manualized United Voices and finalized the design of a pilot randomized controlled trial to evaluate the feasibility and acceptability of the intervention protocol and procedures. The findings provide insights into the barriers and facilitators involved in culturally tailoring interventions for older people living with HIV, implementing intervention adaptations within web-based environments, and the promise of developing hybrid music-based interventions for older adults with HIV., (©Miranda Hill, Meredith Greene, Julene K Johnson, Judy Y Tan. Originally published in JMIR Formative Research (https://formative.jmir.org), 08.10.2024.)

Pollinator-plant interactions represent a core mutualism that underpins biodiversity in terrestrial ecosystems, and the loss of flowering plants is a major driver of pollinator declines. Bee attraction to flowers is mediated by both quantity of resources (the number of available flowers for exploration) and quality of resources (pollen nutritional value), but whether and how bees prioritize these factors is not well understood. Here, we leveraged a unique plant system to investigate the floral factors influencing bee foraging decisions. Recombinant inbred plant lines were generated by crossing the self-fertilizing Capsella rubella and the pollinator-dependent outcrosser C. grandiflora , to produce plants that varied across floral traits. Using enclosed arenas, we evaluated the foraging behavior of two solitary bee species, Osmia cornifrons and Megachile rotundata , to the isolated inflorescences from these lines. Visits from O. cornifrons were significantly positively correlated with the number of flowers, while M. rotundata visits were significantly positively associated with pollen nutrition, with a preference for plants with higher pollen protein-to-lipid content. Further experiments using artificial flowers confirmed that M. rotundata preferred flowers with higher protein:lipid ratios, while O. cornifrons visits were unaffected by nutrition. These studies demonstrate that, although both bee species collect pollen as their sole source of protein and lipids for themselves and/or their offspring, they differentially prioritize resource quantity (number of flowers) and quality (pollen nutritional content). These studies lay the groundwork for understanding how different foraging strategies evolved, and influence, plant-pollinator ecological networks., (© The Author(s) 2024. Published by Oxford University Press on behalf of National Academy of Sciences.)

Objectives: Induction of labor is commonly undertaken when ongoing pregnancy poses a risk to either the mother or fetus. Often cervical preparation is required with mechanical methods increasingly popular due to their improved safety. This study evaluates the efficacy, safety, and acceptability of digital versus speculum-based balloon insertion for cervical preparation, aiming to identify gaps and inform future research., Data Source: PubMed, Ovid MEDLINE, EMBASE, and Scopus were searched from database inception until 30 June 2023., Study Eligibility Criteria: Included studies were randomized controlled trials comparing digital versus speculum-based insertion of catheter-related balloons for labor induction in individuals with viable singleton pregnancies, in both inpatient and outpatient settings, written in English. Exclusions included studies not using cervical balloons, comparisons to nonballoon methods, nonhuman studies, and nonprimary literature like guidelines, reviews, commentaries, and opinion pieces., Methods: Title and abstract screening were performed by 4 authors. Full-text articles were assessed against inclusion criteria. Selection was agreed upon by consensus among 3 authors, with a fourth consulted for disputes. The risk of bias was assessed using the Cochrane Risk of Bias Tool 2.0 for randomized trials. A meta-analysis was also performed., Results: Out of 3397 studies, 4 met the inclusion criteria, all being randomized controlled trials with some concerns in at least one domain but no high risk of bias. Two studies found digital insertion significantly less painful than speculum-based insertion (P<.001), while one reported no difference (P=.72). Maternal satisfaction was comparable, with one study favoring digital insertion (P=.011). Meta-analysis findings for other outcome measures suggest no difference between speculum or digital insertion. However, due to substantial heterogeneity, findings for procedural time, time from induction to delivery, and epidural rate should be cautiously interpreted., Conclusions: Digital insertion for cervical preparation appears associated with reduced pain and higher patient acceptability compared to speculum-based insertion. Additionally, efficacy and safety were comparable, indicating it is a preferable option for clinical use. There was no difference in other procedural, obstetric, or neonatal outcomes, however, more rigorous research employing standardized outcome measures is needed to facilitate a clinically meaningful interpretation., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Prenatal sequencing tests are being introduced into clinical practice in many developed countries. In part due to its greater ability to detect genetic variation, offering prenatal sequencing can present ethical challenges. Here we review ethical issues arising following the implementation of prenatal sequencing in the English National Health Service (NHS). We analysed semi structured interviews conducted with 48 parents offered prenatal sequencing and 63 health professionals involved in delivering the service to identify the ethical issues raised. Two main themes were identified: (1) Equity of access (including issues around eligibility criteria, laboratory analytical processes, awareness and education of clinicians, fear of litigation, geography, parental travel costs, and access to private healthcare), and (2) Timeliness and its impact on parental decision-making in pregnancy (in the context of the law around termination of pregnancy, decision-making in the absence of prenatal sequencing results, and the "importance" of prenatal sequencing results). Recognising both the practical and systemic ethical issues that arise out of delivering a national prenatal sequencing service is crucial. Although specific to the English context, many of the issues we identified are applicable to prenatal sequencing services more broadly. Education of health professionals and parents will help to mitigate some of these ethical issues., (© 2024. The Author(s).)

Purpose: The goal of this study was to develop a lot release assay for iPSC residuals following directed differentiation of iPSCs to retinal pigment epithelial (RPE) cells. Methods: RNA Sequencing (RNA Seq) of iPSCs and RPE derived from them was used to identify pluripotency markers downregulated in RPE cells. Quantitative real time PCR (qPCR) was then applied to assess iPSC residuals in iPSC-derived RPE. The limit of detection (LOD) of the assay was determined by performing spike-in assays with known quantities of iPSCs serially diluted into an RPE suspension. Results: ZSCAN10 and LIN28A were among 8 pluripotency markers identified by RNA Seq as downregulated in RPE. Based on copy number and expression of pseudogenes and lncRNAs ZSCAN10 and LIN28A were chosen for use in qPCR assays for residual iPSCs. Reverse transcription PCR indicated generally uniform expression of ZSCAN10 and LIN28A in 21 clones derived from 8 iPSC donors with no expression of either in RPE cells derived from 5 donor lines. Based on qPCR, ZSCAN10 , and LIN28A expression in iPSCs was generally uniform. The LOD for ZSCAN10 and LIN28A in qPCR assays was determined using spike in assays of RPE derived from 2 iPSC lines. Analysis of ΔΔC t found the limit of detection to be <0.01% of cells, equivalent to <1 iPSC/10,000 RPE cells in both iPSC lines. Conclusions: qPCR for ZSCAN10 and LIN28A detects <1 in 10,000 residual iPSCs in a population of iPSC-derived RPE providing an adequate LOD of iPSC residuals for lot release testing.

Cellular processes are controlled by the thermodynamics of the underlying biomolecular interactions. Frequently, structural investigations use one monomeric binding partner, while ensemble measurements of binding affinities generally yield one affinity representative of a 1:1 interaction, despite the majority of the proteome consisting of oligomeric proteins. For example, viral entry and inhibition in SARS-CoV-2 involve a trimeric spike surface protein, a dimeric angiotensin-converting enzyme 2 (ACE2) cell-surface receptor and dimeric antibodies. Here, we reveal that cooperativity correlates with infectivity and inhibition as opposed to 1:1 binding strength. We show that ACE2 oligomerizes spike more strongly for more infectious variants, while exhibiting weaker 1:1 affinity. Furthermore, we find that antibodies use induced oligomerization both as a primary inhibition mechanism and to enhance the effects of receptor-site blocking. Our results suggest that naive affinity measurements are poor predictors of potency, and introduce an antibody-based inhibition mechanism for oligomeric targets. More generally, they point toward a much broader role of induced oligomerization in controlling biomolecular interactions., Competing Interests: Competing interests statement:P.K. is a nonexecutive director, shareholder of and consultant to Refeyn Ltd., J.L.P.B. and W.B.S. are shareholders of and consultants to Refeyn Ltd. W.B.S. and P.K. received the University of Oxford’s COVID-19 Research Response Fund. P.K. has filed a patent for the contrast enhancement methodology and its application to mass measurement of single biomolecules. G.R.S. is on the GSK Vaccines Scientific Advisory Board, a founder shareholder of RQ biotechnology and Jenner investigator. Oxford University holds intellectual property related to the Oxford-AstraZeneca vaccine.

Background: Pneumonia accounts for over half a million older adult emergency department (ED) visits annually, but ED pneumonia diagnosis is inaccurate. Geriatric-specific pneumonia diagnostic criteria exist for other settings; no prospective data exist to determine if application in the older adult ED population is feasible. The objective was to prospectively evaluate the utility of four current diagnostic criteria (Loeb; Modified McGeer; Infectious Disease Society of America/American Thoracic Society; American College of Emergency Physicians) in older adult ED patients., Methods: This was a prospective, observational cohort study of older adult ED patients ≥65 years of age in two U.S. EDs with suspected pneumonia defined as having chest radiography ordered and treating physician suspicion. The standard we used for defining the presence, absence, or inability to determine a diagnosis of pneumonia diagnosis was expert physician chart adjudication. We report the summary statistics for demographic characteristics and symptoms/exam findings and sensitivity, specificity, and likelihood ratios with 95% confidence intervals of the existing diagnostic criteria. Pre-specified cutoff values of a positive LR >10 and a negative LR <0.3 were considered clinically significant., Results: Of 135 patients enrolled, 27 had pneumonia by adjudicator review. Typical patient-reported pneumonia symptoms, such as fever (18.5%) and new/worse cough (51.9%), were not consistently present in pneumonia. The IDSA/ATS and ACEP criteria had positive LR >10 and negative LR <0.3; however, all confidence intervals included pre-specified cutoffs., Conclusions: Older adults presented to the ED with low frequency of typical pneumonia symptoms. Although existing diagnostic definitions had promising test characteristics, they may not perform well enough for clinical application without refinement., (© 2024 The Author(s). Journal of the American Geriatrics Society published by Wiley Periodicals LLC on behalf of The American Geriatrics Society.)

Competing Interests: Declaration of Interests K.H. has nothing to disclose. D.B. has nothing to disclose. N.W. has nothing to disclose. E.G. has nothing to disclose. M.H. has nothing to disclose. S.T. has nothing to disclose. The views expressed in this manuscript are those of the author(s) and do not necessarily reflect the official policy or position of the Defense Health Agency, Department of Defense, or the US Government.

Genome sequencing (GS) has the potential to reduce the "diagnostic odyssey" that many parents of children with rare undiagnosed conditions experience. While much research has considered the impact of receiving a diagnostic result, research has rarely focused solely on the impact of receiving a "no primary finding" (NPF) result. This study aimed to investigate the experience of parents of children with rare and undiagnosed conditions following an NPF result from GS. Nine parents whose child had an NPF result from GS were recruited through the social media platform of the charity SWAN (Syndromes Without A Name) UK. Semi-structured telephone interviews were conducted, transcribed verbatim, and analyzed using grounded theory. Analysis led to the emergence of two main themes. The first theme "Striving to Solve the Unsolved Puzzle" concerned the experience of striving to end the "diagnostic odyssey." The second theme "Navigating Hope, Lost then Found" plots the trajectory of hope raised by the promise of a new technology, dashed by the NPF, and the eventual return of small and distant hope for the future. Taken together, these themes allowed for a proposed theory: "The Disequilibrium of Hope," which highlights the dynamic and modifiable experience of hope participants experience in their GS journey. These results suggest GS can be an emotional rollercoaster for parents. While hope plays an important role in coping with the day-to-day life of living with a rare disease, careful management of expectations from GS is important during pre-test counseling, and continued follow-up and support are needed beyond result disclosure. An understanding of the disappointment and distress caused by an NPF result is valuable for healthcare professionals in this field to ensure counseling can be tailored. Further research should consider how to support parents after an NPF result., (© 2023 The Authors. Journal of Genetic Counseling published by Wiley Periodicals LLC on behalf of National Society of Genetic Counselors.)

Background: With the potential to identify a vast number of rare diseases soon after birth, genomic newborn screening (gNBS) could facilitate earlier interventions and improve health outcomes. Designing a gNBS programme will involve balancing stakeholders' opinions and addressing concerns. The views of medical students-future clinicians who would deliver gNBS-have not yet been explored., Methods: We conducted a nationwide online survey of UK medical students via the REDCap platform. Perceptions of gNBS, including scope of testing and potential benefits and drawbacks, were explored using a mix of multiple-choice questions, Likert scales, visual analogue scales and free-text questions., Results: In total, 116 medical students across 16 universities participated. Overall, 45% supported gNBS, with a positively skewed mean support score of 3.24 (SD 1.26, range: 1.0-5.0), and 55% felt it relevant to their future practice. Almost all agreed that infant-onset and childhood-onset diseases and conditions with effective treatments should be included. Most felt that earlier interventions and personalised care would be the most important benefit of gNBS. Other perceived benefits included earlier diagnoses, diagnosing more patients and enabling research for new treatments. However, several perceived challenges were highlighted: risk of genomic discrimination, incidental or uncertain findings, data security and breaching children's future autonomy. Students expressed conflicting opinions on the psychological impact on families, but most were concerned about a lack of support due to current resource limitations in health services. Students frequently reported having insufficient knowledge to form an opinion, which may reflect gaps in genomics education at medical school and the current lack of evidence base for gNBS., Conclusion: Although some support for gNBS was demonstrated, ethicolegal and social challenges were raised, emphasising a need for ongoing discussions about the implications of gNBS., Competing Interests: Competing interests: AP is an employee of Genomics England, a company wholly owned by the United Kingdom Department of Health and Social Care and funded to deliver the Generation Study., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Black women continue to be disproportionally burdened by HIV. Pre-exposure prophylaxis (PrEP) is an effective HIV prevention option, which is underused by Black women. While social network interventions (SNIs) have been widely researched and implemented among some groups vulnerable to HIV, little is known about social network characteristics among Black women. To learn more about the social networks of Black women vulnerable to HIV and their knowledge of and interest in PrEP, we conducted a cross-sectional survey among 109 Black women aged 18-45 years attending a family planning clinic in Chicago, Illinois. In our study, 44% of women reported that they were moderately to extremely concerned about HIV. Over half of participants (53%) had a small personal network size (i.e., less than two). No statistically significant associations between having larger network sizes and having previously heard of PrEP, having an interest in starting PrEP, or having good PrEP knowledge were detected. Open-ended responses revealed high levels of trust in network connections with matters related to sexual health. Additionally, nearly all (94%) of women reported that SNIs were a good idea to promote PrEP. Future network studies are needed to inform the development of effective intervention strategies for women., Competing Interests: The authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Background: Neoadjuvant endocrine therapy presents an important downstaging option with lower toxicity than neoadjuvant chemotherapy in oestrogen receptor (ER)-positive early breast cancer. Meta-analysis of the effects of neoadjuvant endocrine therapy on surgical outcomes across randomized clinical trials (RCTs) and cohort studies has not previously been performed., Methods: A systematic review and meta-analysis was performed to evaluate the effect of neoadjuvant endocrine therapy on surgical outcomes (PROSPERO (international prospective register of systematic reviews, 2020)) compared with surgery followed by adjuvant endocrine therapy. PubMed and EMBASE were searched to identify RCT and cohort studies between 1946 and 27 March 2024. Two independent reviewers manually screened the identified records and extracted the data. Risk of bias was assessed using the Cochrane Collaboration tools and random-effects meta-analysis was done with ReviewManager., Results: The search identified 2390 articles eligible for screening. The review included 20 studies (12 cohort and 8 RCTs); 19 were included in the meta-analysis with a total of 6382 patients. Overall, neoadjuvant endocrine therapy was associated with a lower mastectomy rate compared with surgery first (risk ratio (RR) 0.53, 95% c.i. 0.44 to 0.64). Subgroup analysis showed similar improvement in the mastectomy rate in the neoadjuvant endocrine therapy group versus control group irrespective of study type (RCT: RR 0.58, 95% c.i. 0.50 to 0.66; cohorts: RR 0.48, 95% c.i. 0.33 to 0.70). There was no difference in the mastectomy rate by duration of neoadjuvant endocrine therapy (more than 4 months: RR 0.57, 95% c.i. 0.42 to 0.78; 4 months or less than 4 months: RR 0.52, 95% c.i. 0.43 to 0.64). Most of the studies were characterized by moderate-quality evidence with significant heterogeneity., Conclusion: Neoadjuvant endocrine therapy is associated with a reduction in mastectomy rate. Given the moderate methodological quality of previous studies, further RCTs are required., Registration Id: CRD42020209257., (© The Author(s) 2024. Published by Oxford University Press on behalf of BJS Foundation Ltd.)

Gene therapy using recombinant adeno-associated virus (rAAV) as delivery vehicles has garnered much interest in recent years. There are still significant gaps in our fundamental understanding of the manufacturing processes to deliver sufficient products. Manufacturing efforts of rAAV using HEK293 cells have commonly relied on fixed bed falling film bioreactors like the iCELLis®. We used computational fluid dynamics (CFD) to validate the operating conditions required for a predictive iCELLis® 500 scale-down model. The small-scale and at-scale systems have different flow paths causing validation of the corresponding agitation rates required to achieve the same linear flow through the fixed bed across scales to be non-trivial. Therefore, we used CFD to predict the theoretical scaling relationship. In addition, CFD could predict kLa differences between the two systems and the operating conditions required to match kLa between scales. We also confirmed that the location of DO control must be the same in both systems to achieve proper scaling. Experimental runs confirming the validity of the novel scale-down model showed that based on the modifications to the iCELLis® Nano system, we achieved similar DO, key metabolite, pH, and GC titer trends in both systems., (© 2024 Wiley Periodicals LLC.)

The global rise of oropharyngeal cancers (OPC) associated with the human papillomavirus (HPV) type 16 necessitates a deeper understanding of their underlying molecular mechanisms. Our study utilised RNA-sequencing data from The Cancer Genome Atlas (TCGA) to identify and analyse differentially expressed (DE) long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and messenger RNAs (mRNAs) in HPV16-positive OPC, and to elucidate the interplay within the lncRNA/miRNA/mRNA regulatory network. We revealed 1929 DE lncRNAs and identified a significant expression shift in 37 of these, suggesting a regulatory 'sponge' function for miRNAs that modulate cellular processes. Notably, the lncRNA Linc00911 exhibited decreased expression in HPV16-positive OPC, a change directly attributable to HPV oncogenes E6 and E7 as confirmed by RT-qPCR in cell lines and patient samples. Our comprehensive analysis presents an expansive landscape of ncRNA-mRNA interactions, offering a resource for the ongoing pursuit of elucidating the molecular underpinnings of HPV-driven OPC., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Stress and negative mood in pregnancy have been linked to less favorable birth outcomes, but the mechanisms underlying this effect remain largely unknown. We examined associations between emotions in pregnancy, pro- and anti-inflammatory cytokines (IFN-γ, IL-6, IL-8, IL-10, IL-12, IL-17, MCP-1, MIP-1β, TNF-α) and birth outcomes (gestational age at birth and birth weight) in a low-risk sample., Methods: At each trimester of pregnancy, participants (N = 74) completed the Positive and Negative Affect Schedule, Perceived Stress Scale, Edinburgh Postnatal Depression Scale, and State-Trait Anxiety Inventory. They provided blood samples in the third trimester. Multivariate regression with a reduction of dimensionality (orthogonal projection to latent structures) was used to assess associations between maternal emotions, cytokine levels, and birth outcomes., Results: We found significant positive associations between negative mood (depressive symptoms in the second and third trimesters and negative affect in the third trimester) and anti-inflammatory cytokine IL-10 levels, and negative associations between maternal distress in the second and third trimesters and pro-/anti-inflammatory cytokine ratios (IFN-γ/IL-10, TNF-α/IL-10 and IL-6/IL-10). Higher levels of pro-inflammatory cytokines IFN-γ, IL-12, IL-17, and TNF-α were associated with younger gestational age at birth and lower birth weight., Limitations: We did not control for relevant factors such as social support, health-related behaviors, or cortisol levels., Conclusions: Negative mood in mid- and late pregnancy may shift cytokine balance toward the anti-inflammatory cytokine dominance. Our results provide further evidence for the negative association between pro-inflammatory cytokines in late pregnancy and gestational age at birth/birth weight, which we observed even in a low-risk population., Competing Interests: Declaration of competing interest None., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Substrate composition has been widely recognised as a primary variable shaping lotic macroinvertebrate communities at the habitat unit level. However, fundamental understanding of how communities inhabiting mineralogical habitats (i.e., gravel, sand and silt) are structured across differing rivers is lacking. Moreover, research largely focusses on gravel beds and fine sediment in general (<2 mm) and as a result detailed field observations specifically of the sand and silt fractions are lacking. Using data from five UK streams collated from published studies, we assess taxonomic and functional biodiversity (alpha and beta diversity) at the habitat unit level (as defined by substrate composition of sand, silt and gravel). We found that the composition of taxonomic communities were clearly different in all habitat units for each individual stream (and at the landscape scale), with comparable, but less strong, distinctions between substrates for functional macroinvertebrate community composition. However, alpha diversity metrics and Local Contribution to Beta Diversity (LCBD) recorded among the different habitat units varied significantly across individual rivers, and the amount of variation explained by the habitat unit for taxonomic and functional composition demonstrated considerable differences suggesting strong context dependence. The depositional fine sediment habitats of sand and silt were found to support a discrete community composition and differing levels of alpha and beta diversity within and between rivers. We advocate that care should be taken when seeking to generalise biodiversity patterns at a landscape scale as our study highlights the high degree of context dependency when considering the role of the habitat template. Moreover, our results provide evidence that discriminating between the size fractions of fine sediment habitats (sand or silt) is important to fully elucidate the wider ecological importance of these habitats and the distinct taxonomic and functional biodiversity they support., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Ecology and Evolution published by John Wiley & Sons Ltd.)

Schizophrenia is associated with numerous abnormalities, including imbalances in all hormonal axes, among which steroids play a major role. Steroidomic studies therefore represent a promising tool for early diagnosis and appropriate treatment of schizophrenia. A total of 51 adult male schizophrenics aged 27 (22, 34) years (shown as median with quartiles) and 16 healthy controls (HCs) aged 28 (25, 32) years were enrolled into this study. Our results showed the effective differentiation of men with schizophrenia from controls based on steroidomic profiles. We also found an altered metabolic pathway from pregnenolone and its sulfate (PREG/S) to cortisol in schizophrenics with several metabolic bottlenecks such as lower PREG levels due to increased PREG sulfation and/or suppressed PREGS desulfation and attenuated conversion of 17-hydroxy-PREG to 17-hydroxy-progesterone, as well as the results suggestive of suppressed CYP11B1 activity. In contrast, steroid molar ratios suggested two counterregulatory steps involving increased conversion of PREG/S to 17-hydroxy-PREG/S and decreased conversion of cortisol to cortisone, which may maintain unchanged basal cortisol levels but may not ensure a sufficient cortisol response to stress. Our data also indicated a trend to higher 7α-, 7β-, and 16α-hydroxylation that may counteract the autoimmune complications and proinflammatory processes accompanying schizophrenia. Finally, a possible suppression of HSD17B3 activity was suggested, resulting in decreased circulating testosterone levels with increased androstenedione levels.

Objective: To study cycle outcomes of women who choose to pursue oocyte cryopreservation, using published age-specific oocyte recommendations., Design: Retrospective cohort., Setting: Clinic., Patient(s): A total of 5,915 patients seeking planned oocyte cryopreservation, 3,504 ultimately underwent ovarian stimulation with oocyte retrieval and cryopreservation, 425 of this cohort subsequently thawed with intent to use embryo transfer., Intervention(s): Planned oocyte cryopreservation., Main Outcome Measure(s): Planned oocyte cryopreservation consultation, fertility preservation cycle(s) and ovarian stimulation outcomes., Result(s): A total of 5,915 women were seen in the initial consultation for planned fertility preservation (2012-2022). Ethnicity and the highest level of education were significant in predicting who would move forward with oocyte stimulation for fertility preservation. Women who reported working within the law and public policy and in the fields of health and medicine were statistically more likely to proceed with a cycle than those who listed other occupations. Of 3,504 women in the study cohort who underwent ovarian stimulation and egg retrieval, 1,331 (38.0%) achieved the age-based recommended number of oocytes to freeze. Only 57 (4.3%) of these women who met their age-based oocyte goal did so after their initial cryopreservation cycle. There was a significant association between ethnicity and number of cryopreservation cycles, specifically showing that Black or African American women were less likely to complete two or more cycles. Patients whose education background included graduate or professional degrees were more likely to have completed more than one cycle., Conclusion(s): Self-identified ethnicity was significantly associated with the odds of moving forward with oocyte stimulation for fertility preservation and egg retrieval after initial consultation with ethnic minorities significantly less likely to continue treatment. Of those who undergo egg freezing, most women pursue more than one stimulation and cryopreservation cycle, yet the majority never meet their recommended number to freeze., Competing Interests: Declaration of Interests D.B. has nothing to disclose. M.B. has nothing to disclose. A.C.C. has nothing to disclose. R.Y. has nothing to disclose. S.R. has nothing to disclose. K.D. reports consulting fees from BluDiagnostics and Medscape; honoraria from University of California, San Diego and Pacific Coast Reproductive Society; travel support from The American Society for Reproductive Medicine, The Society for Assisted Reproductive Technology and U.S. Fertility; data safety monitoring boards for Medscape, BluDiagnostics, Future Fertility, and LifeWhisperer; The Society for Reproductive Endocrinology and Infertility Research Chair, The Society for Assisted Reproductive Technology QA Chair, Medical Director; and Chief Research Officer, U.S. Fertility; stocks in U.S. Fertility (indirect) and Shady Grove Fertility; options in LifeWhisperer and Future Fertility. M.H. has nothing to disclose. A.D. has nothing to disclose. T.S. has nothing to disclose., (Published by Elsevier Inc.)

Debridement is an important component of wound management and can improve outcomes for patients. Debridement needs to be done by an appropriately trained health professional, but the scope of practice, credentials, training, competencies, and regulatory requirements regarding wound debridement can differ. Best Practice Recommendations were created to positively influence patient safety related to all methods of debridement, across the continuum of care, and to be implemented widely by nurses at all professional levels in Canada., Aim: To further develop the Best Practice Recommendations for wound debridement, with an international perspective, by creating a consensus document to support the global adoption of evidence-based debridement practice for health professionals., Methods: A consensus meeting utilising Delphi methods was conducted between the authors to review the consensus statements. Once 80 % agreement was achieved, a wide range of wound care experts were identified by the authors and invited to participate in an external review of the statements., Results: Fifteen consensus statements about wound debridement were agreed upon and are presented in this paper., Conclusions: These best practice recommendations have been reviewed by a wide range of practitioners from across the UK and Canada and aim to provide guidance on the standardisation of debridement practices for healthcare professionals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Maximum likelihood estimation is among the most widely-used methods for inferring phylogenetic trees from sequence data. This paper solves the problem of computing solutions to the maximum likelihood problem for 3-leaf trees under the 2-state symmetric mutation model (CFN model). Our main result is a closed-form solution to the maximum likelihood problem for unrooted 3-leaf trees, given generic data; this result characterizes all of the ways that a maximum likelihood estimate can fail to exist for generic data and provides theoretical validation for predictions made in Parks and Goldman (Syst Biol 63(5):798-811, 2014). Our proof makes use of both classical tools for studying group-based phylogenetic models such as Hadamard conjugation and reparameterization in terms of Fourier coordinates, as well as more recent results concerning the semi-algebraic constraints of the CFN model. To be able to put these into practice, we also give a complete characterization to test genericity., (© 2024. The Author(s).)

We have previously identified a parasite-derived peptide, FhHDM-1, that prevented the progression of diabetes in nonobese diabetic (NOD) mice. Disease prevention was mediated by the activation of the PI3K/Akt pathway to promote β -cell survival and metabolism without inducing proliferation. To determine the molecular mechanisms driving the antidiabetogenic effects of FhHDM-1, miRNA:mRNA interactions and in silico predictions of the gene networks were characterised in β -cells, which were exposed to the proinflammatory cytokines that mediate β -cell destruction in Type 1 diabetes (T1D), in the presence and absence of FhHDM-1. The predicted gene targets of miRNAs differentially regulated by FhHDM-1 mapped to the biological pathways that regulate β -cell biology. Six miRNAs were identified as important nodes in the regulation of PI3K/Akt signaling. Additionally, IGF-2 was identified as a miRNA gene target that mediated the beneficial effects of FhHDM-1 on β -cells. The findings provide a putative mechanism by which FhHDM-1 positively impacts β -cells to permanently prevent diabetes. As β -cell death/dysfunction underlies diabetes development, FhHDM-1 opens new therapeutic avenues., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2024 Inah Camaya et al.)

A patient in her 30s who was a G0 proceeded with in vitro fertilisation (IVF) for a history of male factor infertility. Following single embryo transfer, the patient was diagnosed with a conjoined twin pregnancy. During her IVF cycle, the patient was stimulated with an antagonist protocol for 13 days followed by a gonadotropin-releasing hormone agonist trigger. 13 eggs were retrieved, 9 were mature and 5 fertilised with intracytoplasmic sperm injection. Of those, two were cryopreserved. She had a successful frozen blastocyst embryo transfer. The patient's 7-week ultrasound demonstrated a single gestational sac, yolk sac and fetal pole. However, the crown-rump length appeared visually abnormal and two heartbeats were visualised. She was referred to maternal-fetal medicine (MFM) for a first-trimester ultrasound. Her ultrasound with MFM was notable for a fluid-filled chest, foreshortened limbs and early sacral agenesis. She was subsequently diagnosed with cephalopagus twins and underwent an induced abortion following consultation with MFM., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2024. No commercial re-use. See rights and permissions. Published by BMJ.)

Introduction: Barriers to seeking infertility care for lesbian, gay, bisexual, transgender, intersex, queer/questioning, and asexual (LGBTIQA+) individuals are well documented in the literature. However, little is known about military LGBTIQA+ service members seeking infertility care within the Military Health System. Approximately 6.1% of active duty U.S. service members across all branches identify as LGBTIQA+, which underscores the need for a deeper understanding of the needs of this community to support and retain service members. We therefore sought to describe the lived experiences of lesbian and gay cisgender service members in building their families in order to understand their family-building desires and potential barriers to seeking infertility care., Materials and Methods: We developed a survey to investigate the impact of military service on family planning. After Institutional Review Board approval, we distributed the survey throughout Walter Reed National Military Medical Center's obstetrics and gynecology clinic and posted the survey on multiple open and closed social media pages for LGBTIQA+ service members. We reported descriptive statistics of our survey and compared binary variables using the Fisher exact test. Following completion of this survey, participants could self-select to participate in semi-structured interviews., Results: Sixty-eight respondents completed our survey and self-identified as either cis-male (n = 28) or cis-female (n = 40). Most respondents (67.9% cis-males, 92.5% cis-females) plan to build their families during their military commitment; however, approximately half (50.0% cis-male, 42.5% cis-female) reported a lack of support in this endeavor. Many respondents were unaware of resources that would assist in the pursuit of donor egg, donor sperm, or surrogacy (78.6% cis-males, 50.0% cis-females). Thirty-six participants elected to complete a follow-up interview. After coding the interviews, 5 themes emerged: (1) barriers to initiating care; (2) institutional barriers within the military; (3) political barriers; (4) knowledge sharing; and (5) implicit and explicit bias., Conclusions: Our results suggest significant barriers to LGBTIQA+ service members seeking infertility care. Overall, LGBTIQA+ service members did not feel supported by the military in building their families. Although the military has expanded access to infertility services, efforts to raise awareness and build support for LGBTIQA+ service members are warranted., (© The Association of Military Surgeons of the United States 2024. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site–for further information please contact journals.permissions@oup.com.)

Objectives: To measure the effect of routine vaccination on serum amyloid A (SAA) concentration in apparently healthy horses. We hypothesized that routine vaccination would increase SAA in healthy horses., Animals: 21 apparently healthy client-owned horses and 15 Kansas State University College of Veterinary Medicine-owned horses., Methods: In experiment 1 (n = 8 horses), a blinded, randomized, prospective, crossover study was performed. Horses were either vaccinated (rabies, tetanus, West Nile, Eastern and Western equine encephalomyelitis, equine herpesvirus-1/-4, influenza) or administered saline, and SAA was measured at 6, 12, and 24 hours and daily until day 10 with a commercial lateral-flow immunoassay. In experiment 2 (n = 28 horses), a prospective, observational study measured SAA after vaccination at 12 and 24 hours and daily until day 10. A linear mixed-effect model with repeated measures over time blocked by horse tested the effect of treatment on SAA. A repeated-measures correlation tested the correlation between SAA and temperature., Results: Over time, vaccinated horses had increased model-adjusted SAA compared to unvaccinated horses without clinical evidence of adverse reaction (P < .01). In experiment 1, the model-adjusted SAA after vaccination peaked on day 2 (median, 1,872 µg/mL; IQR, 1,220.8 to 2,402.5 µg/mL) and returned to normal (< 20 µg/mL) by day 9 (median, 6 µg/mL; IQR, 0.8 to 23.5 µg/mL) after vaccination. In experiment 2, vaccinated horses had increased SAA over time; temperature and SAA were not correlated (P = .78)., Clinical Relevance: Results of this study indicated that routine vaccination results in increased SAA concentration and provided evidence for a period of convalescence following vaccination. Measuring SAA for 10 days following vaccination cannot be used as an indicator of illness.

Contrast-enhanced mammography is being increasingly implemented clinically, providing much improved contrast between tumour and background structures, particularly in dense breasts. Although CEM is similar to conventional mammography it differs via an additional exposure with high energy X-rays (≥ 40 kVp) and subsequent image subtraction. Because of its special operational aspects, the CEM aspect of a CEM unit needs to be uniquely characterised and evaluated. This study aims to verify the utility of a commercially available phantom set (BR3D model 020 and CESM model 022 phantoms (CIRS, Norfolk, Virginia, USA)) in performing key CEM performance tests (linearity of system response with iodine concentration and background subtraction) on two models of CEM units in a clinical setting. The tests were successfully performed, yielding results similar to previously published studies. Further, similarities and differences in the two systems from different vendors were highlighted, knowledge of which may potentially facilitate optimisation of the systems., (© 2024. Crown.)

Objective: Concerned allies often call crisis lines and other call centers about their loved ones' mental health. Callers to Coaching Into Care, a non-crisis call center, often report concerns about suicide risk in Veterans yet little is known about how best to support those callers. We conducted a documentation review to understand standard operating procedures, barriers, and opportunities for risk reduction., Method: Across 1,581 unique callers with an initial call over a 6 month period, 225 callers (14.2%) were identified for contact note review and coding., Results: Calls were frequently characterized by current suicidal ideation (62.7%), suicide attempts (24.0%), and access to lethal means (35.6%), although use of lethal means safety interventions was infrequently documented (12.9%). The majority of callers were coded as open to intervention (83.4%). After coaching, 16% of Veterans who were previously not connected to mental health care had connected to care in the community or VA., Conclusions: There was substantial heterogeneity in assessments and interventions used, particularly related to documentation of access to lethal means; however, there is an opportunity to provide risk reduction education and communication skill building for family and friends of Veterans. Those closest to Veterans report being open to learning new ways to engage with Veterans around safety and mental wellness.

Background: The Antimicrobial Resistance Laboratory Network (AR Lab Network) was developed by the CDC to detect emerging antimicrobial-resistant (AR) threats and prevent outbreaks. However, low submission rates of AR isolates limit the potential of the AR Lab Network to address antimicrobial resistance (AMR)., Aim: The aim of this study was to investigate barriers to submission of AR isolates in acute care hospitals (ACHs) and critical access hospitals (CAHs) within Texas Public Health Region 8 (PHR8) counties., Methods: A survey was designed and emailed to laboratory professionals to identify barriers to AR isolate submission. Responses were analyzed using 2-sided Fisher's exact tests to identify associations between responses and respondent characteristics., Results: Of the 33 hospitals within PHR8 invited to participate in the survey, responses were received from 21, a response rate of 63.6%. Lack of awareness of the AR Lab Network was the most frequently cited barrier to submission (65.4% of respondents). Other reported barriers to submission included lack of laboratory staff time (57.7%), lack of training with the submission process (34.6%), lack of personnel certified to ship infectious substances (23.1%), and lack of laboratory/shipping supplies (23.1%)., Discussion: Regardless of the respondent's role, time in that role, or type of hospital in which they worked, the most common barrier to isolate submission was lack of awareness of the AR Lab Network. In the future, we will address the identified barriers by implementing educational outreach programs about AMR and the AR Lab Network for hospitals and other healthcare facilities within PHR8., Competing Interests: The author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: Bailie Moorhead, Niva Shrestha, Alvin Boyd Newman-Caro, Sydney L. Vangeli, Victoria N. Lussier, Mark I. Grijalva, Margaret E. Giro, Danielle Natividad Jackson, Tianna M. Mack, Zackary L. Herrle, Marcheta Hill, Gretchen Rodriguez, Rachael Singer, Cynthia Williams, Adaugo Nwanguma, Enyinnaya N. Merengwa, Rachel Pittman, and Rebecca L. Sanchez declare no conflicts of interest., (© The Author(s) 2024.)