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Matthew B Miller,1 Melissa J Roumanis,1 Lisa Kakinami,2,3 Geoffrey C Dover1,3,4 1Department of Health, Kinesiology, and Applied Physiology, Concordia University, Montreal, Canada; 2Department of Mathematics & Statistics, Concordia University, Montreal, Canada; 3PERFORM Centre, Concordia University, Montreal, Canada; 4Centre de Recherché Interdisciplinaire en Réadaptation du Montréal Metropolitain, Montreal, CanadaCorrespondence: Matthew B MillerDepartment of Health, Kinesiology, and Applied Physiology, Concordia University, L - SP 165.28, Richard J. Renaud Science Complex, 7141 Sherbrooke W, Montreal, CanadaTel +1 514-848-2424 ext. 3304Fax +1 514-848-8681Email ma_iller@live.concordia.caPurpose: To examine the relationship between baseline kinesiophobia and baseline pain catastrophizing with the 4-day average activity intensity at different times of the day while accounting for different wake and sleep-onset times in chronic pain patients.Methods: Twenty-one participants suffering from idiopathic chronic pain completed baseline questionnaires about kinesiophobia, catastrophizing, disability, depression, and pain. We measured the participants’ activity using accelerometers and calculated activity intensity in the morning, afternoon, and evening. We performed a 2-way repeated measures ANOVA to compare activity levels at different times of the day, and multiple linear regressions.Results: Baseline kinesiophobia was significantly associated with 4-day average evening light activity and sedentary activity at all time periods while baseline catastrophizing was significantly associated with increased 4-day average light activity in the evening and more moderate to vigorous activity in the morning. Our participants engaged in more light activity on average than sedentary activity, and very little moderate-vigorous activity. Participants were most active in the afternoon.Conclusion: Baseline kinesiophobia and baseline catastrophizing were not associated with the 4-day average total daily activity; however, they were associated with 4-day average activity intensities at different times throughout the day. Segmenting daily activity into morning, afternoon, evening may influence the relationship between daily activity, and kinesiophobia and pain catastrophizing. Individuals with chronic pain are less sedentary than previously thought which may affect future interventions.Keywords: accelerometer, physical activity, pain, Tampa Scale, sedentary, daily activity

Category: Ankle Arthritis; Ankle; Arthroscopy Introduction/Purpose: Early ankle arthritis can present with pain and anterior impingement. Younger, active patients who have exhausted conservative treatment have two options; ankle arthrodesis or ankle arthroplasty. Both are major procedures that have pros and cons. Revision surgery for both of these procedures is a major undertaking. Therefore, fit active patients who are under 65, may benefit from a less invasive procedure of anterior ankle arthroscopy and debridement. There is a risk, however that debridement alone, may inadvertently lead to the acceleration of joint degeneration. Thus the arthroscopic insertion of an acellular allograft dermal matrix acts as a 'cushion' to the joint surface and 'buy some time' for the patient to delay major surgery while having a reduction in their symptoms of pain and stiffness. Methods: Patients had surgery performed as a day case under general anaesthetic and tourniquet. No joint distraction was used. Two anterior portals were utilised; anetermedial and anterolateral. A 4mm arthroscope was used to visualise the joint surface. A arthroscopic shave and burr were used to remove any scar soft tissue and tibial and talar osteophytes. Any lateral ankle instability of the ATFL ligament, confirmed, clinically and with MRI imaging, was addressed with stabilisation using arthroscopic placement of an augmented stabilization with synthetic brace. A piece of acellular allograft dermal matrix was cut to size and place through one of the anterior portals onto the joint surface of the talus. The allograft was held in place with tisseal glue. Post-operatively the patient was place in a moonboot which was worn day and night for two weeks, then day only for a further two weeks, non weight- bearing. Three months of physiotherapy followed. Results: The majority of patients had an improvement in their range of motion. Most had reduction in their symptoms of pain. Patients who were less active appeared to have a longer duration of benefit versus those who were keen to return to full activity as quickly as possible. There was no incidence of infection or septic arthritis. One allograft tranlated anteriorly out of the joint space, resulting in a degree of impingement. Patients who wanted to consider further surgery presented on average two to three years after the index procedure. Conclusion: Anterior ankle arthroscopy with arthroscopic debridement of osteophytes and acellular allograft dermal matrix interposition, is a more minor procedure than ankle arthrodesis or ankle arthroplasty. This provides patients with an alternative to major surgery at a time in their lives where they wish to delay major intervention. The most challenging aspect of the procedure is inserting the acellular allograft dermal matrix through a small anterior portal and inserting it into the joint surface.

Category: Diabetes Introduction/Purpose: 2020-2022 saw the COVID-19 coronavirus pandemic. Stratification of access to theatres based on urgency resulted in many diabetic patients 'falling through the crack'. Diabetic patients with foot disease, who presented acutely and/or with sepsis were often managed by the vascular team, who often deemed that there was no salvage option available. However, patients who had recurrent ulceration but were systemically well were unable to access the theatre and clinical skills required to address the mechanical deformity that was responsible for the recurrence of the ulcer. Methods: Scotland National Database for all diabetic foot disease collates information for all new diabetic foot ulcers. The system is designed to collate data pertaining to the control of the patients' diabetes, peripheral vascular disease and smoking. It is less robust had assessing orthopaedic and biomechanical abnormalities which would benefit from surgical intervention. A collaborative approach with the diabetologists, diabetic podiatrists and orthotics, orthopaedic and vascular surgeons across Scotland has anecdotally recorded a significant reduction in services for this group of patients. Community podiatry services were redeployed during the pandemic. Access to hospital clinics and theatre utilisation was stopped for periods of time. The very first weeks had patients staying at home with loved ones and as a result, active ulcers decreased due to off-loading and patients taking care of themselves. Recovery however is painfully slow, with disjointed services. It is however an opportunity to reshape a national diabetic foot service. Results: Initial review with all the major stakeholders anecdotally has reported significant increases in diabetic foot disease during the two years of the pandemic. The lack of access to elective orthopaedic procedures has meant that patients are now presenting very late. Often the only surgical option available to save life or limb is amputation. However new ways of working have brought some benefits to patients. Virtual and remote communication has facilitated a reduction in the number of hospital visits. However, lack of coordination means that many of these patients are suboptimally managed pre and peri-operatively. Clinical competencies are very varied across the country, and there has been no face-to-face training. This is however a perfect opportunity to re-think the delivery of service and allocation of training and vital resources across the country. Conclusion: A complete review of the national picture of delivery of services for patients with diabetic foot disease allows identification of patients' needs. The allocation of resources and training needs for both surgeons and Allied Health Professionals will be identified and addressed. The main focus of the review remains the reduction in amputation rates, with an associated reduction in mortality for this group of patients.Once pathways have been reassessed, escalation should become more robust. The review will also identify major training needs.

Category: Diabetes; Bunion; Lesser Toes Introduction/Purpose: Diabetic forefoot ulcers are associated with a high risk of recurrence, subsequent infection and amputation. Patients who have an amputation for diabetic foot disease have a significantly increased 5-year morbidity and mortality. Minimally invasive surgery to redistribute forces includes any combination of percutaneous Achilles/extensor tendon lengthening, osteotomy/excision of the metatarsal head or sesamoids, and phalangeal osteotomy. The purpose of this case- controlled cohort study is to determine if there is any significant reduction in re-ulceration and amputations rates in patients treated with Neuropathic Minimally Invasive Surgeries, versus conservative management with wound debridement, antibiotics and total contact casting. Methods: Patients who had presented to the diabetic foot clinics that subsequently underwent Neuropathic Minimally Invasive Surgery over a 5-year period were matched with a similar group of patients (age, sex, neuropathy, peripheral vascular disease) who were managed conservatively. The conservative group of patients were identified from the Scottish SCI-Diabetes database. Outcomes of recurrent ulceration and amputation, time to heal and number of clinic visit were all compared. Results: Patients who underwent minimally invasive surgery had predictable healing of their ulcers in 6 weeks. Very few represented a recurrent ulcer, though some did develop an ulcer at a different site. Subsequent amputation rates were lower in the surgical group versus the non-surgical group. The number of clinic visits significantly reduced in the operated group, versus the conservatively treated group. Conclusion: Patients with recurrent diabetic foot ulcers benefit from early assessment to determine if they would be a good surgical candidate for Neuropathic Minimally Invasive Surgery. Many diabetic patients have an alteration in their foot biomechanics, whether soft tissue, osseous or both. Minimally invasive surgeries provide relatively predictive times to ulcer healing. Re-ulceration at the same time is reduced. This has a significant impact on the patients' quality of life. There is also a major benefit to healthcare systems where there is a significant burden of the health care budget being allocated for regular dressings and wound reviews.

Category: Diabetes; Midfoot/Forefoot Introduction/Purpose: Charcot neuroarthropathy is associated with a progressive deformity that can result in wound breakdown, ulceration and ultimately amputation. Conservative management with total contact casting can allow ulcers to heal. However conservative treatment is dependent on continual podiatry and orthotic input. Surgical correction of the 'rocker bottom deformity has evolved. Minimally invasive surgical techniques utilising beams and bolts for a 'super-construct' to correct the Charcot deformity is possible through percutaneous incisions. We report the 5-year outcomes of a cohort of patients with diabetic neuroarthropathy treated with neuropathic minimally invasive surgeries including modifications of the surgical technique to reduce complications and improve outcomes. Methods: A cohort of 16 patients with diabetic neuroarthropathy was treated using neuropathic minimally invasive surgeries. Patients were recruited from the multidisciplinary diabetic foot clinic. Patients had been treated with total contact casting to heal ulcers and prevent future wound breakdown. Surgery was performed as a day case or overnight stay under general anaesthetic. Using fluoroscopy, closing wedge osteotomy was performed with minimally invasive 'Shannon' cutting and wedge burrs to create a triplane closing wedge osteotomy of the midfoot. Reduction of the forefoot to the midfoot was achieved using percutaneous beams and bolts. The void was filled with antibiotic bone graft substitute. Post-operatively patients were immobilised in total contact cast for eight to twelve weeks non-weight-bearing, followed by six weight-bearing in a moon-boot with total contact insole for six weeks. Results: The aim of surgical intervention is to create a stable plantigrade shoe able foot. Whilst osseous union is desirable, a stable fibrous union allows the patient to transition to footwear, with reduced risk of recurrence of deformity, wound breakdown or ulceration, Minimally invasive surgery with antibiotic bone graft substitutes reduces the risk of post-operative infection. The most common complication was the breakage of metalwork at the tip of the bolts. Infection was the second most common complication. Removal of metalwork, minimally invasive debridement and irrigation, allowed the infection to be irradicated. Lost of correction of deformity requiring revision surgery occurred in three patients with good outcomes. There was one amputation. Conclusion: Significant midfoot deformity correction can be performed using minimally invasive surgical techniques with beams and bolts to create a super-construct for stabilisation of midfoot Charcot deformity. The advantages of percutaneous incisions allow for earlier intervention before a major deformity has occurred. Neuropathic minimally invasive surgeries are optimal for patients who have no active ulceration and are systemically well but require prophylactic intervention to prevent further deformity and risk of amputation.

Cannabis is the most widely produced and consumed illicit psychoactive substance worldwide. Occasional cannabis use can progress to frequent use, abuse and dependence with all known adverse physical, psychological and social consequences. Individual differences in cannabis initiation are heritable (40-48%). The International Cannabis Consortium was established with the aim to identify genetic risk variants of cannabis use. We conducted a meta-analysis of genome-wide association data of 13 cohorts (N=32 330) and four replication samples (N=5627). In addition, we performed a gene-based test of association, estimated single-nucleotide polymorphism (SNP)-based heritability and explored the genetic correlation between lifetime cannabis use and cigarette use using LD score regression. No individual SNPs reached genome-wide significance. Nonetheless, gene-based tests identified four genes significantly associated with lifetime cannabis use: NCAM1, CADM2, SCOC and KCNT2. Previous studies reported associations of NCAM1 with cigarette smoking and other substance use, and those of CADM2 with body mass index, processing speed and autism disorders, which are phenotypes previously reported to be associated with cannabis use. Furthermore, we showed that, combined across the genome, all common SNPs explained 13-20% (P

We previously reported that tacrolimus (TAC) trough blood concentrations for African American (AA) kidney allograft recipients were lower than those observed in white patients. Subtherapeutic TAC troughs may be associated with acute rejection (AR) and AR-associated allograft failure. This variation in TAC troughs is due, in part, to differences in the frequency of the cytochrome P450 CYP3A5*3 allele (rs776746, expresses nonfunctional enzyme) between white and AA recipients; however, even after accounting for this variant, variability in AA-associated troughs is significant. We conducted a genomewide association study of TAC troughs in AA kidney allograft recipients to search for additional genetic variation. We identified two additional CYP3A5 variants in AA recipients independently associated with TAC troughs: CYP3A5*6 (rs10264272) and CYP3A5*7 (rs41303343). All three variants and clinical factors account for 53.9% of the observed variance in troughs, with 19.8% of the variance coming from demographic and clinical factors including recipient age, glomerular filtration rate, anticytomegalovirus drug use, simultaneous pancreas-kidney transplant and antibody induction. There was no evidence of common genetic variants in AA recipients significantly influencing TAC troughs aside from the CYP3A gene. These results reveal that additional and possibly rare functional variants exist that account for the additional variation.

Recent studies have demonstrated the importance of sex effects on the underlying genetic architecture of insulin-related traits. To explore sex-specific genetic effects on fasting insulin, we tested for genotype-by-sex interaction and conducted linkage analysis of fasting insulin in Hypertension Genetic Epidemiology Network families. Hypertensive siblings and their first-degree relatives were recruited from five field centers. We performed a genome scan for quantitative trait loci influencing fasting insulin among 1,505 European Americans and 1,616 African Americans without diabetes. Sex-stratified linear regression models, adjusted for race, center, and age, were explored. The Mammalian Genotyping Service typed 391 microsatellite markers, spaced roughly 9 cM. Variance component linkage analysis was performed in SOLAR using ethnic-specific marker allele frequencies and multipoint IBDs calculated in MERLIN. We detected a quantitative trait locus influencing fasting insulin in female subjects (logarithm of odds [LOD] = 3.4) on chromosome 2 at 95 cM (between GATA69E12 and GATA71G04) but not in male subjects (LOD = 0.0, P for interaction = 0.007). This sex-specific signal at 2p13.2 was detected in both European-American (LOD = 2.1) and African-American (LOD = 1.2) female subjects. Our findings overlap with several other linkage reports of insulin-related traits and demonstrate the importance of considering complex context-dependent interactions in the search for insulin-related genes. [ABSTRACT FROM AUTHOR]

Objective: Metabolic syndrome-related traits (obesity, glucose intolerance/insulin resistance, dyslipidemia, and hypertension) have been shown to be genetically correlated. It is less clear, however, if the genetic correlation extends to novel risk factors associated with inflammation, impaired fibrinolytic activity, and hyperuricemia. We present a bivariate genetic analysis of MetS-related traits including both traditional and novel risk factors.Research Design and Methods: Genetic correlations were estimated using a variance components procedure in 1,940 nondiabetic white individuals from 445 families in the National Heart, Lung, and Blood Institute (NHLBI) Family Heart Study. Twelve MetS-related traits, including BMI, waist circumference, blood pressure, white blood cell count, fasting serum triglycerides, HDL cholesterol, insulin, glucose, plasminogen activator inhibitor-1 antigen, uric acid, and C-reactive protein, were measured and adjusted for covariates, including lifestyle variables.Results: Significant genetic correlations were detected among BMI, waist circumference, HDL cholesterol, triglycerides, insulin, and plasminogen activator inhibitor-1 antigen and between uric acid and all of the above variables except insulin. C-reactive protein and white blood cell count were genetically correlated with each other, and both showed significant genetic correlations with waist circumference and insulin. Fasting glucose was not significantly genetically correlated with any of the other traits.Conclusions: These results suggest that pleiotropic effects of genes or shared family environment contribute to the familial clustering of MetS-related traits. [ABSTRACT FROM AUTHOR]

Type 2 diabetes susceptibility is determined by multiple genetic and environmental factors. Genome-wide linkage scans have localized common regions, possibly harboring susceptibility genes on chromosomes 1, 2, 12, and 20. Variability in linkage findings underscores the probable genetic heterogeneity of type 2 diabetes. Thus, we conducted a genome scan of diabetes status using maximum likelihood methods that model affection status by a liability threshold model. Hypertensive sibships and their offspring and/or parents in the Hypertension Genetic Epidemiology Network study were recruited from five field centers. The diabetes phenotype was derived using the World Health Organization criteria and adjusted for race/study center, age, age2, sex, and with and without percent body fat. In total, 567 diabetic participants were identified in 437 families. Variance component linkage analysis was performed among 1,545 Caucasians and 1,608 African Americans using race-specific marker allele frequencies. We detected a quantitative trait loci (QTLs) influencing diabetes variance (logarithm of odds = 3.4) on chromosome 22, which overlaps a positive type 2 diabetes finding among Canadian Oji-Cree Indians. We also observed suggestive evidence for linkage on chromosomes 1, 2, 5, 8, 14, 17, and 19. The identification and replication of type 2 diabetes QTLs will bring us closer to the detection of functional genes that influence diabetes susceptibility. [ABSTRACT FROM AUTHOR]

Background: Insomnia symptoms are pervasive and persistent in alcohol use disorder (AUD), though little is known about the mechanisms that underlie this association. We previously found that cognitive behavioral therapy for insomnia (CBT-I) reduced alcohol-related problems among veterans by improving insomnia severity (NCT03806491). In this planned secondary analysis of the same clinical trial data, we tested negative emotionality as one potential mechanism to explain this effect. Specifically, we tested the change in negative emotionality as a mediator of the association between change in insomnia symptoms and alcohol-related outcomes (craving, heavy drinking frequency, and alcohol-related problems)., Methods: Participants were 67 veterans in treatment for AUD who also met the criteria for insomnia disorder (91% male, 84% White, average age = 46.3 years). Participants were randomized to five sessions of CBT-I or a single-session sleep hygiene control. Assessments occurred at baseline, immediately posttreatment (~6 weeks after baseline), and at 6-week follow-up. Measures included the Insomnia Severity Index, Penn Alcohol Craving Scale, Timeline Followback, and Short Inventory of Problems. We created a latent negative emotionality indicator based on five validated and reliable measures of negative emotionality., Results: Contrary to hypotheses, CBT-I did not improve negative emotionality relative to sleep hygiene control. However, across both treatment conditions, decreases in insomnia symptoms from baseline to posttreatment were associated with concurrent decreases in negative emotionality, which in turn predicted reductions in alcohol craving and heavy drinking., Conclusion: Negative emotionality may help explain links between insomnia symptoms and alcohol-related outcomes., (© 2024 Research Society on Alcohol.)

Background: Poor sleep is common in the context of cannabis use, but experimental and daily-life studies provide mixed evidence for whether cannabis use helps or disturbs same-night sleep. Despite a high prevalence of co-use of alcohol and cannabis, most studies of cannabis use and sleep do not consider alcohol use. We sought to add to this literature by examining associations between cannabis and sleep in the context of alcohol co-use and to examine these associations bidirectionally using ecological momentary assessment., Methods: Participants were 88 adults reporting cannabis use at least 3×/week and simultaneous alcohol and cannabis use at least 2×/week. For 14 days, participants completed morning surveys assessing hours slept and perceived sleep quality from the night before. In morning surveys, random surveys, and substance-specific surveys, participants also reported on alcohol and cannabis craving, use, and amounts., Results: Primary results from multilevel models demonstrated that cannabis use was not independently associated with sleep (duration or quality). However, cannabis use attenuated alcohol's negative effects on sleep quality., Conclusions: Results question the utility of cannabis use to improve sleep but highlight the attenuated negative effects of alcohol as a potential reinforcer of alcohol-cannabis co-use. Future work should continue to consider polysubstance use and integrate additional self-report and objective measures of sleep health to further clarify how cannabis use affects sleep., (© 2024 Research Society on Alcohol.)

Objectives: To evaluate causes of fever, including resistance patterns, in patients undergoing cancer treatment in Malawi., Methods: In this prospective cohort study, enrolled patients undergoing chemotherapy at Kamuzu Central Hospital in Lilongwe, Malawi were given a thermometer. If a temperature of ≥38°C was recorded, they were instructed to return for hospitalization, standardized fever workup, and antibiotics. All patients were followed through 90 days post-fever event or completion of chemotherapy., Results: One hundred and five patients were screened and 50 were enrolled. Of the enrollees, 26 (52%) were men and 26 (52%) were HIV positive, with a mean ART duration of 7 years and CD4 count of 293 cells/µL. The most common diagnoses were aggressive non-Hodgkin lymphoma (40%) and Hodgkin lymphoma (22%). Twenty-three febrile events were recorded from 15 patients. Of the 23 events, a causative agent was isolated in 13 cases: Escherichia coli (6), Plasmodium falciparum (3), Streptococcus pneumoniae (2), Pseudomonas aeruginosa (1) and Citrobacter freundii (1). Of the six E. coli isolates, all were found to be resistant to fluoroquinolones and 4/6 (66%) were resistant to cephalosporins. All patients but one survived; the death was attributed to Pseudomonas bacteraemia., Conclusions: This study describes laboratory-confirmed causes of neutropenic fever (NF) in cancer patients in Malawi. Gram-negative rods, followed by malaria, were the most common source of infection. Gram-negative rods were associated with high rates of antimicrobial resistance. Malaria and resistant bacterial infections should be considered for NF treatment and prevention in sub-Saharan Africa., (© The Author(s) 2024. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

Importance: Anti-β-amyloid immunotherapy using lecanemab is becoming increasingly available to patients with Alzheimer disease (AD). Individuals with Down syndrome (DS) develop AD neuropathology by age 40 years, representing a significant cohort of genetically determined AD., Objective: To investigate the binding properties of lecanemab in the brains of people with DS, in anticipation of their inclusion in clinical trials or access to antiamyloid immunotherapies., Design, Setting, Participants: The study included cases of postmortem brain tissue analysis from 15 individuals with DS aged 43 to 68 years that were acquired from Alzheimer Disease research centers at the University of California, Irvine and the University of Kentucky from 2008 to 2021. Data were analyzed from August 2023 through May 2024., Exposure: The binding properties of lecanemab were assessed in brain tissue., Main Outcome: The primary outcome was the extent of lecanemab binding to amyloid plaques and brain blood vessels., Results: Tissue from 15 people (8 were female [53%]) with DS ranging in age from 43 to 68 (mean, 56.6) years were included in the study. Lecanemab-labeled amyloid plaques appeared in all 15 DS cases studied, indicating potential target engagement. However, extensive binding of lecanemab to brain blood vessels in DS was observed, raising significant safety concerns. These findings underscore the necessity for clinical trials of lecanemab in people with DS to evaluate both safety and efficacy, particularly in individuals older than 43 years., Conclusions and Relevance: These findings suggest significant binding of lecanemab to cerebral amyloid angiopathy in DS. Lecanemab should be rigorously tested in clinical trials for AD in the DS population to determine its safety and efficacy, especially in those older than 43 years.

Creutzfeldt-Jakob Disease (CJD), the most common human prion disease, is associated with pathologic misfolding of the prion protein (PrP), encoded by the PRNP gene. Of human prion disease cases, < 1% were transmitted by misfolded PrP, ~ 15% are inherited, and ~ 85% are sporadic (sCJD). While familial cases are inherited through germline mutations in PRNP, the cause of sCJD is unknown. Somatic mutations have been hypothesized as a cause of sCJD, and recent studies have revealed that somatic mutations accumulate in neurons during aging. To investigate the hypothesis that somatic mutations in PRNP may underlie sCJD, we performed deep DNA sequencing of PRNP in 205 sCJD cases and 170 age-matched non-disease controls. We included 5 cases of Heidenhain variant sporadic CJD (H-sCJD), where visual symptomatology and neuropathology implicate localized initiation of prion formation, and examined multiple regions across the brain including in the affected occipital cortex. We employed Multiple Independent Primer PCR Sequencing (MIPP-Seq) with a median depth of > 5000× across the PRNP coding region and analyzed for variants using MosaicHunter. An allele mixing experiment showed positive detection of variants in bulk DNA at a variant allele fraction (VAF) as low as 0.2%. We observed multiple polymorphic germline variants among individuals in our cohort. However, we did not identify bona fide somatic variants in sCJD, including across multiple affected regions in H-sCJD, nor in control individuals. Beyond our stringent variant-identification pipeline, we also analyzed VAFs from raw sequencing data, and observed no evidence of prion disease enrichment for the known germline pathogenic variants P102L, D178N, and E200K. The lack of PRNP pathogenic somatic mutations in H-sCJD or the broader cohort of sCJD suggests that clonal somatic mutations may not play a major role in sporadic prion disease. With H-sCJD representing a localized presentation of neurodegeneration, this serves as a test of the potential role of clonal somatic mutations in genes known to cause familial neurodegeneration., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Introduction: Transcranial direct current stimulation (tDCS) administers low-intensity direct current electrical stimulation to brain regions via electrodes arranged on the surface of the scalp. The core promise of tDCS is its ability to modulate brain activity and affect performance on diverse cognitive functions (affording causal inferences regarding regional brain activity and behavior), but the optimal methodological parameters for maximizing behavioral effects remain to be elucidated. Here we sought to examine the effects of 10 stimulation and experimental design factors across a series of five cognitive domains: motor performance, visual search, working memory, vigilance, and response inhibition. The objective was to identify a set of optimal parameter settings that consistently and reliably maximized the behavioral effects of tDCS within each cognitive domain., Methods: We surveyed tDCS effects on these various cognitive functions in healthy young adults, ultimately resulting in 721 effects across 106 published reports. Hierarchical Bayesian meta-regression models were fit to characterize how (and to what extent) these design parameters differentially predict the likelihood of positive/negative behavioral outcomes., Results: Consistent with many previous meta-analyses of tDCS effects, extensive variability was observed across tasks and measured outcomes. Consequently, most design parameters did not confer consistent advantages or disadvantages to behavioral effects-a domain-general model suggested an advantage to using within-subjects designs (versus between-subjects) and the tendency for cathodal stimulation (relative to anodal stimulation) to produce reduced behavioral effects, but these associations were scarcely-evident in domain-specific models., Discussion: These findings highlight the urgent need for tDCS studies to more systematically probe the effects of these parameters on behavior to fulfill the promise of identifying causal links between brain function and cognition., 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. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Santander, Leslie, Li, Skinner, Simonson, Sweeney, Deen, Miller and Brunye.)

DNMT3A mutations are frequently found in clonal hematopoiesis and a variety of hematologic malignancies, including acute myeloid leukemia. An assortment of mouse models have been engineered to explore the tumorigenic potential and malignant lineage bias due to loss of function of DNMT3A in consort with commonly comutated genes in myeloid malignancies, such as Flt3, Nras, Kras, and c-Kit. We employed several tamoxifen-inducible Cre-ER T2 murine model systems to study the effects of constitutively active Kras G12D -driven myeloid leukemia (Kras) development together with heterozygous (3aHet) or homozygous Dnmt3a deletion (3aKO). Due to the rapid generation of diverse nonhematologic tumors appearing after tamoxifen induction, we employed a transplantation model. With pretransplant tamoxifen induction, most Kras mice died quickly of T-cell malignancies regardless of Dnmt3a status. Using posttransplant induction, we observed a dose-dependent effect of DNMT3A depletion that skewed the leukemic phenotype toward a myeloid lineage. Specifically, 64% of 3aKO/Kras mice had exclusively myeloid disease compared with 36% of 3aHet/Kras and only 13% of Kras mice. Here, 3aKO combined with Kras led to increased disease burden, multiorgan infiltration, and faster disease progression. DOT1L inhibition exerted profound antileukemic effects in malignant 3aKO/Kras cells, but not malignant cells with Kras mutation alone, consistent with the known sensitivity of DNMT3A-mutant leukemia to DOT1L inhibition. RNAseq from malignant myeloid cells revealed that biallelic Dnmt3a deletion was associated with loss of cell-cycle regulation, MYC activation, and TNF⍺ signaling. Overall, we developed a robust model system for mechanistic and preclinical investigations of acute myeloid leukemia with DNMT3A and Ras-pathway lesions., Competing Interests: Conflicts of Interest Disclosure The authors declare no relevant competing financial interests., (Copyright © 2024 International Society for Experimental Hematology. Published by Elsevier Inc. All rights reserved.)

The human cerebral cortex, pivotal for advanced cognitive functions, is composed of six distinct layers and dozens of functionally specialized areas 1,2 . The layers and areas are distinguished both molecularly, by diverse neuronal and glial cell subtypes, and structurally, through intricate spatial organization 3,4 . While single-cell transcriptomics studies have advanced molecular characterization of human cortical development, a critical gap exists due to the loss of spatial context during cell dissociation 5,6,7,8 . Here, we utilized multiplexed error-robust fluorescence in situ hybridization (MERFISH) 9 , augmented with deep-learning-based cell segmentation, to examine the molecular, cellular, and cytoarchitectural development of human fetal cortex with spatially resolved single-cell resolution. Our extensive spatial atlas, encompassing 16 million single cells, spans eight cortical areas across four time points in the second and third trimesters. We uncovered an early establishment of the six-layer structure, identifiable in the laminar distribution of excitatory neuronal subtypes by mid-gestation, long before the emergence of cytoarchitectural layers. Notably, while anterior-posterior gradients of neuronal subtypes were generally observed in most cortical areas, a striking exception was the sharp molecular border between primary (V1) and secondary visual cortices (V2) at gestational week 20. Here we discovered an abrupt binary shift in neuronal subtype specification at the earliest stages, challenging the notion that continuous morphogen gradients dictate mid-gestation cortical arealization 6,10 . Moreover, integrating single-nuclei RNA-sequencing and in situ whole transcriptomics revealed an early upregulation of synaptogenesis in V1-specific Layer 4 neurons, suggesting a role of synaptogenesis in this discrete border formation. Collectively, our findings underscore the crucial role of spatial relationships in determining the molecular specification of cortical layers and areas. This work not only provides a valuable resource for the field, but also establishes a spatially resolved single-cell analysis paradigm that paves the way for a comprehensive developmental atlas of the human brain., Competing Interests: Competing interests C.A.W. has stock ownership in Maze Therapeutics, and is a paid consultant for Third Rock Ventures and Flagship Pioneering. Mingyao Li receives research funding from Biogen Inc., unrelated to the current manuscript. Mingyao Li is a cofounder of OmicPath AI LLC. The remaining authors declare no competing interests.

Urea, as one of the most sustainable organic solutes, denies the high salt consumption in commercial electrolytes with its peculiar solubility in water. The bi-mixture of urea-H 2 O shows the eutectic feature for increased attention in aqueous Zn-ion electrochemical energy storage (AZEES) technologies. While the state-of-the-art aqueous electrolyte recipes are still pursuing the high-concentrated salt dosage with limited urea adoption and single-anion selection category. Here, a dual-anion urea-based (DAU) electrolyte composed of dual-Zn salts and urea-H 2 O-induced solutions is reported, contributing to a stable electric double-layer construction and in situ organic/inorganic SEI formation. The optimized ZT 2 S 0.5 -20U electrolytes show a high initial Coulombic efficiency of 93.2% and durable Zn-ion storage ≈4000 h regarding Zn//Cu and Zn//Zn stripping/plating procedures. The assembled Zn//activated carbon full cells maintain ≈100% capacitance over 50 000 cycles at 4 A g -1 in coin cell and ≈98% capacitance over 20 000 cycles at 1 A g -1 in pouch cell setups. A 12 × 12 cm 2 pouch cell assembly illustrates the practicality of AZEES devices by designing the cheap, antifreezing, and nonflammable DAU electrolyte system coupling proton donor-acceptor molecule and multi-anion selection criteria, exterminating the critical technical barriers in commercialization., (© 2024 The Authors. Small published by Wiley‐VCH GmbH.)

The severe acute respiratory syndrome coronavirus 2 pandemic is characterized by the emergence of novel variants of concern (VOCs) that replace ancestral strains. Here, we dissect the complex selective pressures by evaluating variant fitness and adaptation in human respiratory tissues. We evaluate viral properties and host responses to reconstruct forces behind D614G through Omicron (BA.1) emergence. We observe differential replication in airway epithelia, differences in cellular tropism, and virus-induced cytotoxicity. D614G accumulates the most mutations after infection, supporting zoonosis and adaptation to the human airway. We perform head-to-head competitions and observe the highest fitness for Gamma and Delta. Under these conditions, RNA recombination favors variants encoding the B.1.617.1 lineage 3' end. Based on viral growth kinetics, Alpha, Gamma, and Delta exhibit increased fitness compared to D614G. In contrast, the global success of Omicron likely derives from increased transmission and antigenic variation. Our data provide molecular evidence to support epidemiological observations of VOC emergence., Competing Interests: Declaration of interests R.S.B. is a member of advisory boards for VaxArt, Takeda, and Invivyd; has consulted for Gilead; and has collaborative projects with Gilead, J&J, and Hillevax focused on unrelated projects., (Copyright © 2024. Published by Elsevier Inc.)

Characterizing somatic mutations in the brain is important for disentangling the complex mechanisms of aging, yet little is known about mutational patterns in different brain cell types. Here, we performed whole-genome sequencing (WGS) of 86 single oligodendrocytes, 20 mixed glia, and 56 single neurons from neurotypical individuals spanning 0.4-104 years of age and identified >92,000 somatic single-nucleotide variants (sSNVs) and small insertions/deletions (indels). Although both cell types accumulate somatic mutations linearly with age, oligodendrocytes accumulated sSNVs 81% faster than neurons and indels 28% slower than neurons. Correlation of mutations with single-nucleus RNA profiles and chromatin accessibility from the same brains revealed that oligodendrocyte mutations are enriched in inactive genomic regions and are distributed across the genome similarly to mutations in brain cancers. In contrast, neuronal mutations are enriched in open, transcriptionally active chromatin. These stark differences suggest an assortment of active mutagenic processes in oligodendrocytes and neurons., Competing Interests: Declaration of interests J.G. is now a Merck Research Laboratories (MRL) employee, and no work related to this manuscript was performed at MRL. P.J.P. is a member of the scientific advisory board for Bioskryb Genomics, Inc., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Pneumocystis jirovecii pneumonia (PJP) is a serious and sometimes fatal infection occurring in immunocompromised individuals. High-risk patients include those with low CD4 counts due to human immunodeficiency virus infection and transplant recipients. The incidence of PJP is increasing, and rapid detection of PJP is needed to effectively target treatment and improve patient outcomes. A common method used is an immunofluorescent assay (IFA), which has limitations, including labor costs, low sensitivity, and requirement for expert interpretation. This study evaluates the performance of the DiaSorin Molecular Pneumocystis jirovecii analyte-specific reagent (ASR) in a laboratory-developed test (LDT) for the direct detection of P. jirovecii DNA without prior nucleic acid extraction. Respiratory samples ( n = 135) previously tested by IFA from 111 patients were included. Using a composite standard of in-house IFA and reference lab PJP PCR, the percent positive agreement for the LDT using the DiaSorin ASR was 97.8% (90/92). The negative percent agreement was 97.7% (42/43). The lower limit of detection of the assay was determined to be 1,200 copies/mL in bronchoalveolar lavage fluid. Analytical specificity was assessed using cultures of oropharyngeal flora and common respiratory bacterial and fungal pathogens. No cross-reactivity was observed. Our study suggests that the DiaSorin Pneumocystis ASR accurately detects P. jirovecii DNA and demonstrates improved sensitivity compared to the IFA method., Importance: Our study is unique compared to other previously published studies on the DiaSorin analyte-specific reagent (ASR) because we focused on microbiological diagnostic methods commonly used (immunofluorescent assay) as opposed to pathology findings or reference PCR. In addition, in our materials and methods, we describe the protocol for the use of the DiaSorin ASR as a singleplex assay, which will allow other users to evaluate the ASR for clinical use in their lab., Competing Interests: Melissa Miller is a member of the DiaSorin Molecular Scientific Advisory Board