Isolation and Characterization of Extracellular Vesicles to Activate Retina Regeneration.

Mammals do not possess the ability to spontaneously repair or regenerate damaged retinal tissue. In contrast to teleost fish which are capable of retina regeneration through the action of Müller glia, mammals undergo a process of reactive gliosis and scarring that inhibits replacement of lost neurons. Thus, it is important to discover novel methods for stimulating mammalian Müller glia to dedifferentiate and produce progenitor cells that can replace lost retinal neurons. Inducing an endogenous regenerative pathway mediated by Müller glia would provide an attractive alternative to stem cell injections or gene therapy approaches. Extracellular vesicles (EVs) are now recognized to serve as a novel form of cell-cell communication through the transfer of cargo from donor to recipient cells or by the activation of signaling cascades in recipient cells. EVs have been shown to promote proliferation and regeneration raising the possibility that delivery of EVs could be a viable treatment for visual disorders. Here, we provide protocols to isolate EVs for use in retina regeneration experiments.

Convergent synthesis of bicyclic boronates via a cascade regioselective Suzuki-Miyaura/cyclisation protocol.

Bicyclic boronates have recently emerged as promising candidates to invoke targeted biomolecular interactions, given their selectivity for specific functionalities. Despite this, the general stability of the C-B bond in vivo, for such heterocycles, remains an intractable challenge that can often preclude their utility in drug discovery. To address this challenge, de novo strategies that allow expedient access to strategically substituted boronates, that enable modulation of the C-B bond are urgently required. Herein we disclose an operationally simple, regioselective cross-coupling/cyclisation reaction of easily accessible vicinal boronic esters with 2-halophenols to rapidly forge 3-substituted bicyclic boronates. The utility of the platform was demonstrated via expedient access to Xeruborbactam derivatives, chemoselective manipulation of formed products and the convergent approach to bicyclic boronates with a pendent biomolecular probe.

In Science Journals.

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Sex differences in the adrenal circadian clock: A role for BMAL1 in the regulation of urinary aldosterone excretion and renal electrolyte balance in mice.

Brain and muscle ARNT-Like 1 (BMAL1) is a circadian clock transcription factor that regulates physiological functions. Male adrenal-specific Bmal1 (ASCre/+::Bmal1) KO mice displayed blunted serum corticosterone rhythms, altered blood pressure rhythm, and altered timing of eating, but there is a lack of knowledge in females. This study investigates the role of adrenal BMAL1 in renal electrolyte handling and urinary aldosterone levels in response to low salt in male and female mice. Mice were placed in metabolic cages to measure 12-hour urinary aldosterone after a standard diet and 7 days low salt diet, as well as daily body weight, 12-hour food and water intake, and renal sodium and potassium balance. Adrenal glands and kidneys were collected at ZT0 or ZT12 to measure expression of aldosterone synthesis genes and clock genes. Compared to littermate controls, ASCre/+::Bmal1 KO male and female mice displayed increased urinary aldosterone in response to a low salt diet, although mRNA expression of aldosterone synthesis genes was decreased. Timing of food intake was altered in ASCre/+::Bmal1 KO male and female mice, with a blunted night/day ratio. ASCre/+::Bmal1 KO female mice displayed decreases in renal sodium excretion in response to low salt, but both male and female KO mice had changes in sodium balance that were time-of-day-dependent. In addition, sex differences were found in adrenal and kidney clock gene expression. Notably, this study highlights sex differences in clock gene expression that could contribute to sex differences in physiological functions.

A Conformation-Specific Approach to Native Top-down Mass Spectrometry.

Native top-down mass spectrometry is a powerful approach for characterizing proteoforms and has recently been applied to provide similarly powerful insights into protein conformation. Current approaches, however, are limited such that structural insights can only be obtained for the entire conformational landscape in bulk or without any direct conformational measurement. We report a new ion-mobility-enabled method for performing native top-down MS in a conformation-specific manner. Our approach identified conformation-linked differences in backbone dissociation for the model protein calmodulin, which simultaneously informs upon proteoform variations and provides structural insights. We also illustrate that our method can be applied to protein-ligand complexes, either to identify components or to probe ligand-induced structural changes.

Burnout and turnover risks for healthcare workers in the United States: downstream effects from moral injury exposure.

Moral injury has emerged as a construct of interest in healthcare workers' (HCW) occupational stress and health. We conducted one of the first multidisciplinary, longitudinal studies evaluating the relationship between exposure to potentially morally injurious events (PMIEs), burnout, and turnover intentions. HCWs (N = 473) completed surveys in May of 2020 (T1) and again in May of 2021 (T2). Generalized Linear Models (robust Poisson regression) were used to test relative risk of turnover intentions, and burnout at T2 associated with PMIE exposure, controlling for T1 covariates. At T1, 17.67% reported they had participated in a PMIE, 41.44% reported they witnessed a PMIE and 76.61% reported feeling betrayed by healthcare or a public health organization. In models including all T1 PMIE exposures and covariates, T2 turnover intentions were increased for those who witnessed a PMIE at T1 (Relative Risk [RR] = 1.66, 95% Confidence Interval [CI] 1.17-2.34) but not those that participated or felt betrayed. T2 burnout was increased for those who participated in PMIE at T1 (RR = 1.38, 95%CI 1.03-1.85) but not those that witnessed or felt betrayed. PMIE exposure is highly prevalent among HCWs, with specific PMIEs associated with turnover intentions and burnout. Organizational interventions to reduce and facilitate recovery from moral injury should account for differences in the type of PMIE exposures that occur in healthcare work environments.

GLP-1 receptor agonists are a transformative prehabilitation tool for weight loss in obese patients undergoing elective hernia repair.

BACKGROUND: Obesity is an independent risk factor for complications after abdominal hernia repair. Glucagon-like-peptide-1 (GLP-1) receptor agonists are gaining popularity as pharmacologic weight loss adjuncts and may help patients reach weight loss goals for surgery. We examine our early experience utilizing GLP-1 agonists versus lifestyle modifications alone to achieve weight loss in patients before elective hernia repair. METHODS: This single-center, retrospective review identified obese patients who underwent elective hernia repair from 2014 to 2023. Patients were asked to achieve a BMI ≤ 33 kg/m2 before surgery. Patients who lost weight with GLP-1 therapy in addition to lifestyle changes were compared to a control cohort that achieved similar preoperative weight loss without GLP-1 therapy. Primary outcome was mean time from GLP-1 agonist initiation and initial surgery clinic visit to surgery. Secondary outcomes were 30-day morbidity, mortality, and reoperation rates, and hernia recurrence. RESULTS: Forty-six patients with ventral/incisional, flank, umbilical, parastomal, inguinal, and hiatal hernias were identified (GLP-1 N = 24, control N = 22). 81.8% (N = 18) of controls had a ventral/incisional hernia, compared to 45.8% (N = 11) of GLP-1 patients (p = 0.03). Mean BMI at GLP-1 agonist initiation was similar to mean BMI at initial clinic visit for controls (38.1 ± 4.9 vs 38.2 ± 2.7 kg/m2, p = 0.66). Preoperative mean percentage total weight loss (14.9 ± 7.5 vs 12.4 ± 6.9 kg, p = 0.39) and mean BMI reduction (6.0 ± 3.8 vs 4.9 ± 2.3 kg/m2, p = 0.43) were similar between groups. The mean time from GLP-1 agonist initiation to surgery was significantly shorter than initial clinic visit to surgery for controls (6.3 ± 4.0 vs 14.7 ± 17.6 months, p = 0.03). There was no statistically significant difference in time from initial clinic visit to surgery between groups (7.6 ± 4.4 vs 14.7 ± 17.6 months, p = 0.06). There was no significant difference in 30-day morbidity between groups (8.3 vs 27.3%, p = 0.13). CONCLUSION: GLP-1 agonists accelerate preoperative weight loss for obese hernia patients without negatively impacting postoperative outcomes.

Item response theory-based continuous test norming.

In norm-referenced psychological testing, an individual's performance is expressed in relation to a reference population using a standardized score, like an intelligence quotient score. The reference population can depend on a continuous variable, like age. Current continuous norming methods transform the raw score into an age-dependent standardized score. Such methods have the shortcoming to solely rely on the raw test scores, ignoring valuable information from individual item responses. Instead of modeling the raw test scores, we propose modeling the item scores with a Bayesian two-parameter logistic (2PL) item response theory model with age-dependent mean and variance of the latent trait distribution, 2PL-norm for short. Norms are then derived using the estimated latent trait score and the age-dependent distribution parameters. Simulations show that 2PL-norms are overall more accurate than those from the most popular raw score-based norming methods cNORM and generalized additive models for location, scale, and shape (GAMLSS). Furthermore, the credible intervals of 2PL-norm exhibit clearly superior coverage over the confidence intervals of the raw score-based methods. The only issue of 2PL-norm is its slightly lower performance at the tails of the norms. Among the raw score-based norming methods, GAMLSS outperforms cNORM. For empirical practice this suggests the use of 2PL-norm, if the model assumptions hold. If not, or the interest is solely in the point estimates of the extreme trait positions, GAMLSS-based norming is a better alternative. The use of the 2PL-norm is illustrated and compared with GAMLSS and cNORM using empirical data, and code is provided, so that users can readily apply 2PL-norm to their normative data. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Ultrasound Microvessel Imaging of the Human Placenta Demonstrates Altered Vessel Densities in Fetal Growth Restriction With Vascular and Immune Pathologies: A Pilot Case-Control Study.

OBJECTIVES: Fetal growth restriction (FGR) is commonly associated with placental dysfunction, increasing perinatal morbidity and mortality. Visualizing placental vessels in utero would be advantageous for identifying functional FGR cause and determining proper management strategies. We aimed to utilize high-sensitivity ultrasound microvessel imaging (HUMI) for quantifying placental vessel density (VD) in pregnancies diagnosed with FGR. METHODS: This pilot case-control study enrolled subjects in the third trimester with a diagnosis of FGR (n = 40) and gestational age-matched controls with normal fetal growth (n = 20) at a 2:1 ratio, respectively. The Verasonics Vantage ultrasound system was used to perform HUMI on each participant at one timepoint. Scanning involved randomized singular value decomposition-based clutter filtering to identify the villous tree, followed by step-by-step scanning to acquire 3-dimensional-like data. Mean VD was calculated from three ultrasound measurements per subject. Additional clinical and pathology data were also collected and compared. RESULTS: Sixteen participants were utilized to establish the scanning protocol and 2 met exclusion criteria at delivery. Thus, VD was successfully measured on 42 pregnancies scanned at 35 weeks 5 days on average. In FGR (n = 24), placental VD was significantly reduced compared to controls (P < .01). VD measures were as good at predicting FGR as systolic/diastolic (S/D) ratios (area under the curve 0.86 versus 0.80). In a smaller cohort, VD in placentas with a diagnosis of inflammatory villitis (n = 10) by histology showed an increase in VD compared to those without inflammation (P = .01). Low VD was correlated with increased S/D ratios (P = .03). CONCLUSIONS: HUMI is useful for identifying altered placental vascularization in utero for FGR. VD may be a valuable indicator for placental health and could lead to improved risk stratification methods considering underlying biology.

Bridging big data in the ENIGMA consortium to combine non-equivalent cognitive measures.

Investigators in neuroscience have turned to Big Data to address replication and reliability issues by increasing sample size. These efforts unveil new questions about how to integrate data across distinct sources and instruments. The goal of this study was to link scores across common auditory verbal learning tasks (AVLTs). This international secondary analysis aggregated multisite raw data for AVLTs across 53 studies totaling 10,505 individuals. Using the ComBat-GAM algorithm, we isolated and removed the component of memory scores associated with site effects while preserving instrumental effects. After adjustment, a continuous item response theory model used multiple memory items of varying difficulty to estimate each individual's latent verbal learning ability on a single scale. Equivalent raw scores across AVLTs were then found by linking individuals through the ability scale. Harmonization reduced total cross-site score variance by 37% while preserving meaningful memory effects. Age had the largest impact on scores overall (- 11.4%), while race/ethnicity variable was not significant (p > 0.05). The resulting tools were validated on dually administered tests. The conversion tool is available online so researchers and clinicians can convert memory scores across instruments. This work demonstrates that global harmonization initiatives can address reproducibility challenges across the behavioral sciences.

2'-O-methyltransferase-deficient yellow fever virus: Restricted replication in the midgut and secondary tissues of Aedes aegypti mosquitoes severely limits dissemination.

The RNA genome of orthoflaviviruses encodes a methyltransferase within the non-structural protein NS5, which is involved in 2'-O-methylation of the 5'-terminal nucleotide of the viral genome resulting in a cap1 structure. While a 2'-O-unmethylated cap0 structure is recognized in vertebrates by the RNA sensor RIG-I, the cap1 structure allows orthoflaviviruses to evade the vertebrate innate immune system. Here, we analyzed whether the cap0 structure is also recognized in mosquitoes. Replication analyses of 2'-O-methyltransferase deficient yellow fever virus mutants (YFV NS5-E218A) of the vaccine 17D and the wild-type Asibi strain in mosquito cells revealed a distinct downregulation of the cap0 viruses. Interestingly, the level of inhibition differed for various mosquito cells. The most striking difference was found in Aedes albopictus-derived C6/36 cells with YFV-17D cap0 replication being completely blocked. Replication of YFV-Asibi cap0 was also suppressed in mosquito cells but to a lower extent. Analyses using chimeras between YFV-17D and YFV-Asibi suggest that a synergistic effect of several mutations across the viral genome accompanied by a faster initial growth rate of YFV-Asibi cap1 correlates with the lower level of YFV-Asibi cap0 attenuation. Viral growth analyses in Dicer-2 knockout cells demonstrated that Dicer-2 is entirely dispensable for attenuating the YFV cap0 viruses. Translation of a replication-incompetent cap0 reporter YFV-17D genome was reduced in mosquito cells, indicating a cap0 sensing translation regulation mechanism. Further, oral infection of Aedes aegypti mosquitoes resulted in lower infection rates for YFV-Asibi cap0. The latter is related to lower viral loads found in the midguts, which largely diminished dissemination to secondary tissues. After intrathoracic infection, YFV-Asibi cap0 replicated slower and to decreased amounts in secondary tissues compared to YFV-Asibi cap1. These results suggest the existence of an ubiquitously expressed innate antiviral protein recognizing 5'-terminal RNA cap-modifications in mosquitoes, both in the midgut as well as in secondary tissues.

Genomic Surveillance for Enhanced Healthcare Outbreak Detection and Control.

Background: Current methods are insufficient alone for outbreak detection in hospitals. Real-time genomic surveillance using offers the potential to detect otherwise unidentified outbreaks. We initiated and evaluated the Enhanced Detection System for Healthcare-associated Transmission (EDS-HAT), a real-time genomic surveillance program for outbreak detection and mitigation. Methods: This study was conducted at UPMC Presbyterian Hospital from November 2021 to October 2023. Whole genome sequencing (WGS) was performed weekly on healthcare-associated clinical bacterial isolates to identify otherwise undetected outbreaks. Interventions were implemented in real-time based on identified transmission. A clinical and economic impact analysis was conducted to estimate infections averted and net cost savings. Results: There were 3,921 bacterial isolates from patient healthcare-associated infections that underwent WGS, of which 476 (12.1%) clustered into 172 outbreaks (size range 2-16 patients). Of the outbreak isolates, 292 (61.3%) had an identified epidemiological link. Among the outbreaks with interventions, 95.6% showed no further transmission on the intervened transmission route. The impact analysis estimated that, over the two-year period, 62 infections were averted, with gross cost savings of $1,011,146, and net savings of $695,706, which translates to a 3.2-fold return on investment. Probabilistic sensitivity analysis showed EDS-HAT was cost-saving and more effective in 98% of simulations. Conclusion: Real-time genomic surveillance enabled the rapid detection and control of outbreaks in our hospital and resulted in economic benefits and improvement in patient safety. This study demonstrates the feasibility and effectiveness of integrating genomic surveillance into routine infection prevention practice, offering a paradigm shift in healthcare outbreak detection and control.

In Science Journals.

Highlights from the Science family of journals.

Ten Years of Bipolar Telehealth: Program Evaluation of a Team-Based Telemental Health Clinic.

Objectives: Telemental health via videoconferencing (TMH-V) can overcome many of the barriers to accessing quality mental health care. Toward this end, in 2011, the U.S. Department of Veterans Affairs (VA) established the National Bipolar Disorders TeleHealth (BDTH) Program to provide expert mental health consultation and treatment to Veterans with bipolar spectrum disorders. Methods: Initial analyses of BDTH services suggested that participants had positive changes in quality-of-care indices and clinical outcomes; however, that evaluation was based on a limited sample of both participants and VA medical centers. We were able to confirm and expand upon those early results by using nearly eight times the number of participants and more than twice as many medical centers. Results: For the 2,456 Veterans who completed the intake to our program, there were significant improvements in some of the quality metrics (e.g., lithium use) and a 54% reduction in positive suicide screens (p < 0.05). The Veterans who completed the initial and postprogram assessments (n = 815) reported a 16.6% reduction in manic symptoms (p < 0.001), a 29.3% reduction in depressive symptoms (p < 0.001), and a 21.2% reduction in mood episodes (p < 0.001). Additionally, these Veterans demonstrated significant improvements (p < 0.001) in mental health-related quality of life between the two assessments. Conclusions: These analyses provide further support for the general effectiveness and safety of telemental health via videoconferencing. Future research should examine the generalizability of these findings across various subgroups (e.g., minority patients, patients in rural areas), populations, and health care systems.

Discovery of electrophilic degraders that exploit SNAr chemistry.

Targeted covalent inhibition (TCI) and targeted protein degradation (TPD) have proven effective in pharmacologically addressing formerly 'undruggable' targets. Integration of both methodologies has resulted in the development of electrophilic degraders where recruitment of a suitable E3 ubiquitin ligase is achieved through formation of a covalent bond with a cysteine nucleophile. Expanding the scope of electrophilic degraders requires the development of electrophiles with tempered reactivity that enable selective ligase recruitment and reduce cross-reactivity with other cellular nucleophiles. In this study, we report the use of chemical moieties that enable nucleophilic aromatic substitution (SNAr) reactions in the rational design of electrophilic protein degraders. Appending an SNAr covalent warhead to several preexisting small molecule inhibitors transformed them into degraders, obviating the need for a defined E3 ligase recruiter. The SNAr covalent warhead is versatile; it can recruit various E3 ligases, including DDB1 and CUL4 associated factor 11 (DCAF11), DDB1 and CUL4 associated factor 16 (DCAF16), and possibly others. The incorporation of an SNAr covalent warhead into the BRD4 inhibitor led to the discovery of degraders with low picomolar degradation potency. Furthermore, we demonstrate the broad applicability of this approach through rational functional switching from kinase inhibitors into potent degraders.

The impact of concurrent HIV-infection on women with vulvar cancer: Comparison of clinical characteristics and outcome.

BACKGROUND: HPV-related vulvar cancer is increasing in prevalence, especially in women living with HIV. Treatment of vulva cancer is based on evidence from HPV-independent cancers, which affect older women. The impact of HIV on vulvar cancer characteristics and treatment outcomes needs to be elucidated. PATIENTS AND METHODS: A retrospective observational study compared the clinical characteristics, treatment, and outcomes of 92 HIV-positive and 131 HIV-negative women with vulvar cancer at our institution. Using descriptive statistics, HIV-positive and negative patients were compared and Cox regression models were tested for differences in mortality and recurrence. RESULTS: HIV-positive patients were 20 years younger than HIV-negative patients (p < 0.001). More than 50% of patients presented with advanced stage cancer, however this was independent of HIV-status. Although HIV infection was associated with poorer survival (p = 0.022); rates of cure (p = 0.933) and recurrence rates (p = 0.8) were similar in HIV-positive and negative women. CONCLUSIONS: Vulvar cancer occurs at a much younger age in women living with HIV. Awareness among HIV-positive women and health care providers would lead to diagnosis of vulvar cancer at an earlier stage. Treatment protocols for HPV-related vulvar cancer should not be altered due to HIV status and should take into consideration the young age of the patients.

Physical Health Among Black Immigrants by Region of Birth: A Test of the Racial Context Hypothesis.

OBJECTIVE: We test the Racial Context Hypothesis by examining the association between racial context of origin and five physical health outcomes (self-rated health, activity limitation, functional activity limitation, lifetime hypertension, and lifetime cancer) among U.S.-born Black Americans and Black immigrants in the United States. DESIGN: This cross-sectional study used 2000 through 2018 waves of the National Health Interview Survey (NHIS). Our subsample was limited to adults 18 years of age or older who self-identified as Black and selected a distinct global region of birth if not U.S. born (N = 212,269). We employed zero-order logistic regression models to estimate the relationships between each measure of health and racial context by region of birth. RESULTS: Supporting the Racial Context Hypothesis, we found Black immigrants from racially mixed (Mexico, Central America, the Caribbean, South America) and majority-Black contexts (Africa) had lower odds of being in fair or poor self-rated health [aOR 0.786; 0.616; 0.611], reporting any activity limitation [aOR = 0.537; 0.369; 0.678], reporting functional activity limitation [aOR 0.619; 0.425; 0.678], reporting lifetime hypertension diagnosis [aOR 0.596; 0.543; 0.618], and reporting lifetime cancer diagnosis [aOR 0.771; 0.326; 0.641] compared to U.S.-born Black Americans. After controlling for sociodemographic and socioeconomic covariates, Black immigrants from majority-White contexts (Europe) did not significantly differ from U.S.-born Black Americans on these five physical health measures. CONCLUSION: This study expands our understanding of the "Black immigrant advantage" by showing that Black immigrants from predominantly Black and racially mixed regions rated their health status as poor or fair less often, experienced less activity or functional activity limitations, and had a lower risk of lifetime hypertension and cancer compared to U.S.-born Black Americans. The significant associations persisted even after controlling for sociodemographic and socioeconomic characteristics. Black immigrant health is not homogenous, and the racial context of origin Black immigrants come from has an association with their health outcomes.

A Survey of Why and How Clinicians Change Reinforcers during Teaching Sessions.

Preference assessments are used to make data-based decisions about which stimuli to use as reinforcers but they can be challenging to conduct frequently enough to avoid problems related to momentary shifts in preference and reinforcer efficacy. It remains unclear whether, why, and how clinicians change reinforcers on a momentary basis. Therefore, this study aimed to determine common reasons for, and methods of, changing reinforcers in practice. Most respondents indicated that they often change reinforcers during a session, do so when the client mands for or attends to different stimuli or refuses the current stimulus, and identify the new reinforcer based on recent client behaviors (e.g., mands) or by providing an informal choice between stimuli. Responses did not vary meaningfully based on respondent credentials, client characteristics, or service goals. Implications for clinical practice as well as future research on methods of momentary preference assessment and reinforcer identification are discussed. Supplementary Information: The online version contains supplementary material available at 10.1007/s40617-023-00847-4.

Splicing the Difference: Harnessing the complexity of the transcriptome in hematopoiesis.

Alternative splicing has long been recognized as a powerful tool to expand the diversity of the transcriptome and the proteome. The study of hematopoiesis, from hematopoietic stem cell maintenance and differentiation into committed progenitors to maturation into functional blood cells, has led the field of stem cell research and cellular differentiation for decades. The importance of aberrant splicing due to mutations in cis has been exemplified in thalassemias, resulting from aberrant expression of beta-globin. The simultaneous development of increasingly sophisticated technologies, in particular the combination of multi-color flowcytometric cell sorting with bulk and single cell sequencing, has provided sophisticated insights into the complex regulation of the blood system. The recognition that mutations in key splicing factors drive myeloid malignancies, in particular myelodysplastic syndromes, has galvanized research into alternative splicing in hematopoiesis and its diseases. In this review, we will update the audience on the exciting novel technologies, highlight alternative splicing events and their regulators with essential functions in hematopoiesis, and provide a high-level overview how splicing factor mutations contribute to hematologic malignancies.

Pacifiers and Teething Devices: A Risk for Craniofacial Injury in Children Under 2 Years Old.

Pacifiers and teething devices are commonly introduced during infancy to soothe and comfort. This study aimed to investigate the incidence of emergency room visits for pediatric craniofacial injuries associated with pacifiers and teething devices. All craniofacial injuries due to "Pacifiers or Teething Rings" in children less than 2 years old were retrieved from the National Electronic Injury Surveillance System (NEISS). Age, sex, race, context of injury, cause of injury, and patient disposition were collected. Query revealed an estimated nationwide incidence of 5424 injuries from the years 2012 to 2021, with a mean age of 12.1 months at the time of injury. Most injuries occurred at home (70.2%), often resulting from falls while using the pacifier/teething device (71.0%). An estimated 97.6% of patients were released after emergency room treatment. Education regarding abstaining from pacifier and teething device use during activity may reduce the incidence of these injuries.