Urban heat mitigation by green and blue infrastructure: Drivers, effectiveness, and future needs.

The combination of urbanization and global warming leads to urban overheating and compounds the frequency and intensity of extreme heat events due to climate change. Yet, the risk of urban overheating can be mitigated by urban green-blue-grey infrastructure (GBGI), such as parks, wetlands, and engineered greening, which have the potential to effectively reduce summer air temperatures. Despite many reviews, the evidence bases on quantified GBGI cooling benefits remains partial and the practical recommendations for implementation are unclear. This systematic literature review synthesizes the evidence base for heat mitigation and related co-benefits, identifies knowledge gaps, and proposes recommendations for their implementation to maximize their benefits. After screening 27,486 papers, 202 were reviewed, based on 51 GBGI types categorized under 10 main divisions. Certain GBGI (green walls, parks, street trees) have been well researched for their urban cooling capabilities. However, several other GBGI have received negligible (zoological garden, golf course, estuary) or minimal (private garden, allotment) attention. The most efficient air cooling was observed in botanical gardens (5.0 ± 3.5°C), wetlands (4.9 ± 3.2°C), green walls (4.1 ± 4.2°C), street trees (3.8 ± 3.1°C), and vegetated balconies (3.8 ± 2.7°C). Under changing climate conditions (2070-2100) with consideration of RCP8.5, there is a shift in climate subtypes, either within the same climate zone (e.g., Dfa to Dfb and Cfb to Cfa) or across other climate zones (e.g., Dfb [continental warm-summer humid] to BSk [dry, cold semi-arid] and Cwa [temperate] to Am [tropical]). These shifts may result in lower efficiency for the current GBGI in the future. Given the importance of multiple services, it is crucial to balance their functionality, cooling performance, and other related co-benefits when planning for the future GBGI. This global GBGI heat mitigation inventory can assist policymakers and urban planners in prioritizing effective interventions to reduce the risk of urban overheating, filling research gaps, and promoting community resilience.

Clinical Evaluation of Safety and Efficacy of a Central Current Good Manufacturing Practices Laboratory Produced Autologous Adipose-Derived Stromal Vascular Fraction Cell Therapy Product for the Treatment of Knee Osteoarthritis.

Knee osteoarthritis (KOA) is a prevalent condition characterized by the progressive deterioration of the entire joint and has emerged as a prominent contributor to disability on a global scale. The nature of the disease and its impact on joint function significantly limit mobility and daily activities, highlighting its substantial influence on patients' overall well-being. Stromal vascular fraction (SVF) is a heterogenous, autologous cell product, containing mesenchymal stem cells, derived from the patient's subcutaneous adipose tissue with demonstrated safety and efficacy in the treatment of KOA patients. We conducted a single-arm, open-label, multisite, FDA approved clinical study in Kellgren-Lawrence severity grade 2-4 KOA patients. The cellular product was manufactured from patient-specific lipoaspirate in a centrally located FDA-compliant manufacturing facility. Twenty-nine subjects were treated with a quality tested single intra-articular injection of GMP manufactured SVF. Adverse events, laboratory values, vital signs, and physical examination findings were monitored during the study period. Robust tolerability, without any substantial safety issues, was demonstrated. Knee pain and function, assessed through the Knee Injury and Osteoarthritis Outcome Score (KOOS), demonstrated notable improvements. These positive benefits persisted for up to 12 months, and the majority of participants expressed satisfaction. SVF from each patient was stored in a liquid nitrogen freezer for future clinical treatments. Unique to this study of autologous cells is the shipment of lipoaspirate from the clinic to a central FDA-compliant manufacturing facility for cleanroom-controlled manufacturing. The cell product characterization data demonstrate that this method produces an equivalent product in terms of cell count and viability with the added benefit of further quality assurance testing, including sterility, endotoxin, and flow cytometry, before patient administration. Clinical Trial Registration Number: NCT04043819.

Adverse Childhood Experiences, Acculturation, and Risky Sexual Behaviors in Hispanic Young Adults: Findings from Project RED.

While adverse childhood experiences (ACE) are well-documented predictors of maladaptive behaviors in adulthood, including risky sexual behaviors (RSB), the influence of acculturation in this association remains unknown. Although Hispanics are a rapidly growing population in the United States and are disproportionately affected by adverse sexual health outcomes, there is a paucity of research examining the interplay of ACE, acculturation, and RSB in this population. We observed the ACE-RSB association and how this relationship varies across U.S. and Hispanic acculturation levels, in a sample of Hispanic young adults (n = 715). Data for this study were from Project RED, a longitudinal study of Hispanic health. We ran regression models to test associations between ACE (0, 1-3, 4+) and several RSB (e.g., early sexual initiation (≤14 years), condomless sex, lifetime sexual partners, and alcohol/drug use before intercourse), and assessed moderation by U.S./Hispanic acculturation. Compared with those without ACE, individuals with 4 + ACE had higher odds of early sexual initiation (AOR: 2.23), alcohol/drug use before last intercourse (AOR: 2.31), and condomless sex (AOR: 1.66), as well as a higher number of lifetime sexual partners (ß: 0.60). For those reporting 4 + ACE, high U.S. acculturation was protective in the association between ACE and using alcohol/drugs before intercourse. Future research implications are discussed.

Crop bromide concentrations following methyl bromide fumigation for pale cyst nematode in southeastern Idaho.

Methyl bromide (MeBr) is a sterilizing fumigant used to control quarantine pests that is restricted due to its detrimental atmospheric effects. The degradation of injected MeBr produces crop-available Br- . Up to five applications of MeBr were used in southeastern Idaho fields to combat the pale cyst nematode (Globodera pallida). Data regarding the uptake and partitioning of Br- in crops following MeBr application in the region were unavailable. Research determined background concentrations of Br- in alfalfa (Medicago sativa L.), barley (Hordeum vulgare L.), corn (Zea mays L.), potato (Solanum tuberosum L.), and wheat (Triticum aestivum L.) compared to MeBr-treated fields. Background Br- concentrations ranged from nondetectable (ND) to 33.0 mg Br- kg-1 ; vegetative tissue concentrations were greater than reproductive, except corn where there was no difference. Nearly all crops grown in MeBr-treated fields had greater Br- concentrations than background. Background-baled-alfalfa tissue Br- concentration was 33.0 mg kg-1 compared to 117.8 mg Br- kg-1 from a MeBr-treated field. Br- concentration in green alfalfa decreased from 79.8 to 36.5 mg Br- kg-1 at the final cutting in a MeBr-treated field, where time after application decreased crop Br- concentrations. Small grains had low Br- concentrations in reproductive tissue (1.7 mg Br- kg-1 ) compared to vegetative tissue (106.5 mg Br- kg-1 ). Corn stover concentration (12.7 mg Br- kg-1 ) was low relative to small-grain straw, but corn ear (5.8 mg Br- kg-1 ) was greater than small-grain reproductive tissue in the MeBr-treated field. Crop selection following MeBr applications should consider the likelihood of elevated Br- concentration for the plant fractions intended end use.

Proximity to Cannabis Retailers and Recent Cannabis Use among a Diverse Sample of California Adolescents.

Background: As of May 2023, 23 states and Washington, DC have legalized the sale of cannabis for adults aged 21+, and 38 states, three territories, and D.C. have legalized medical cannabis. Exposure to cannabis retailers could increase adolescent cannabis use. Few studies have examined the impact of residential proximity to cannabis retailers on adolescent cannabis use, and previous findings are inconsistent. Methods: This study examined associations between proximity to cannabis retailers and past 30-day cannabis use. Data were from Project Cal Teens, a statewide survey of California adolescents regarding cannabis-related opinions/behaviors (N = 1406, mean age = 15.5 years, 48% female, 38% Hispanic/Latinx, 33% White, 15% Asian/Pacific Islander, 10% Black/African American, 5% Other/Multiracial). Participants were recruited via schools/social media in 2018-2020. Results: For every additional 5 driving miles to the nearest cannabis retailer, the risk of past 30-day cannabis use was reduced by 3.6% [IRR: 0.964; 95% CI: 0.935-0.994]. For every additional 5 miles from a licensed retailer, the risk of past 30-day cannabis use was reduced by 4.3% [IRR: 0.957; 95% CI: 0.926-0.989]. Interaction analyses revealed that among Hispanic/Latinx students, every 5 miles from a licensed retailer was associated with an 11.9% reduction in the risk of past 30-day cannabis use [IRR: 0.881; 95% CI: 0.820-0.945]. Conclusions: As the number of cannabis retailers in the U.S. increases with the continued legalization of adult cannabis use, research examining the effects of these policies on underage use is crucial. Interventions could include enforcement of zoning laws in/near residential areas to reduce accessibility of adolescents to cannabis retailers.

Surveying Adolescents During a Pandemic: Comparison of Adolescents Recruited via Social Media vs. Schools.

School-based surveys of adolescents can be logistically difficult and exclude students who do not attend school. Social media recruitment could be a promising strategy to recruit representative samples of adolescents. However, few studies have compared adolescent survey data collected via different methodologies. Our team was conducting a school-based survey when the COVID-19 pandemic closed all schools, necessitating a shift to online adolescent recruitment. To achieve our goal of obtaining a sample of high school students throughout California, we placed ads on social media. We compared the adolescents recruited in schools with those recruited on social media on demographic characteristics, mental health, and substance use. The sample of students recruited in schools (N = 737) and adolescents recruited via social media (N = 953) did not differ significantly on gender or substance use. However, compared with school-based recruitment, social media recruitment yielded a higher proportion of boys, whites, and Asians and a lower proportion of girls, Hispanic/Latinx adolescents, and those who spoke other languages at home. The social media sample had significantly higher levels of depression and anxiety symptoms and perceived stress than the school-based sample. Results indicate that social media can be useful for recruiting adolescents for survey research, especially if strategies such as Spanish-language social media ads are used to recruit and consent Hispanic/Latinx adolescents and those with non-English-speaking parents. This method could potentially replace school-based surveys in cases where schools are unwilling to participate in research, or it could be used to supplement school-based samples. Advantages and disadvantages of both methods are discussed.

E-Cigarette-Related Health Beliefs Expressed on Twitter Within the U.S.

Introduction: This mixed-methods study analyzed English-language U.S.-based Twitter posts related to E-cigarette use from February 2021. Methods: Posts were manually identified as health-related or not and, if health-related, whether they were posted by an E-cigarette user. A random selection of 1,000 health-related tweets from 986 unique E-cigarette users were qualitatively content analyzed for theory of planned behavior constructs as well as nature and tone of each tweet message. Using quantitative semantic network analysis, relationships among the identified topics and sentiment-specific conversation patterns were explored. Results: The most salient health-related conversation topics of E-cigarette users, health beliefs corresponding to each theory of planned behavior construct, and major motivational contexts of E-cigarette use were identified. Seven topics emerged in positive tweets: smoking cessation, social impact generation, controls over addiction, therapeutic effects on physical and mental health, social support, device attachment, and peer influence. Nine topics emerged in negative tweets: side effects on physical health, vaping addiction, lack of E-cigarette regulations, peer pressure, increased risk of COVID-19, side effects on mental health, no help in smoking cessation, social conflict, and polysubstance use. Most assertions for E-cigarette benefits were not substantiated. Jokes in tweets appeared to contribute to the view of vaping as an attractive, enjoyable, safe, and fun activity. Discussions about positive aspects of E-cigarette use were concentrated on a few related topics, whereas tweets discouraging E-cigarette use presented a diverse, less related set of topics. Conclusions: The results provide insights into the drivers of E-cigarette use behaviors. E-cigarette user perspectives gathered from social media may inform research to guide future prevention and cessation interventions.

Three-Dimensional-Printed Titanium Versus Polyetheretherketone Cages for Lumbar Interbody Fusion: A Systematic Review of Comparative In Vitro, Animal, and Human Studies.

Interbody fusion is a workhorse technique in lumbar spine surgery that facilities indirect decompression, sagittal plane realignment, and successful bony fusion. The 2 most commonly employed cage materials are titanium (Ti) alloy and polyetheretherketone (PEEK). While Ti alloy implants have superior osteoinductive properties they more poorly match the biomechanical properties of cancellous bones. Newly developed 3-dimensional (3D)-printed porous titanium (3D-pTi) address this disadvantage and are proposed as a new standard for lumbar interbody fusion (LIF) devices. In the present study, the literature directly comparing 3D-pTi and PEEK interbody devices is systematically reviewed with a focus on fusion outcomes and subsidence rates reported in the in vitro, animal, and human literature. A systematic review directly comparing outcomes of PEEK and 3D-pTi interbody spinal cages was performed. PubMed, Embase, and Cochrane Library databases were searched according to PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines. Mean Newcastle-Ottawa Scale score for cohort studies was 6.4. A total of 7 eligible studies were included, comprising a combination of clinical series, ovine animal data, and in vitro biomechanical studies. There was a total population of 299 human and 59 ovine subjects, with 134 human (44.8%) and 38 (64.4%) ovine models implanted with 3D-pTi cages. Of the 7 studies, 6 reported overall outcomes in favor of 3D-pTi compared to PEEK, including subsidence and osseointegration, while 1 study reported neutral outcomes for device related revision and reoperation rate. Though limited data are available, the current literature supports 3D-pTi interbodies as offering superior fusion outcomes relative to PEEK interbodies for LIF without increasing subsidence or reoperation risk. Histologic evidence suggests 3D-Ti to have superior osteoinductive properties that may underlie these superior outcomes, but additional clinical investigation is merited.

Preparing for the next outbreak: A review of indices measuring outbreak preparedness, vulnerability, and resilience.

The COVID-19 pandemic has highlighted the need for relevant metrics describing the resources and community attributes that affect the impact of communicable disease outbreaks. Such tools can help inform policy, assess change, and identify gaps to potentially reduce the negative outcomes of future outbreaks. The present review was designed to identify available indices to assess communicable disease outbreak preparedness, vulnerability, or resilience, including articles describing an index or scale developed to address disasters or emergencies which could be applied to addressing a future outbreak. This review assesses the landscape of indices available, with a particular focus on tools assessing local-level attributes. This systematic review yielded 59 unique indices applicable to assessing communicable disease outbreaks through the lens of preparedness, vulnerability, or resilience. However, despite the large number of tools identified, only 3 of these indices assessed factors at the local level and were generalizable to different types of outbreaks. Given the influence of local resources and community attributes on a wide range of communicable disease outcomes, there is a need for local-level tools that can be applied broadly to various types of outbreaks. Such tools should assess both current and long-term changes in outbreak preparedness with the intent to identify gaps, inform local-level decision makers, public policy, and future response to current and novel outbreaks.

A novel porcine model of CLN3 Batten disease recapitulates clinical phenotypes.

Mouse models of CLN3 Batten disease, a rare lysosomal storage disorder with no cure, have improved our understanding of CLN3 biology and therapeutics through their ease of use and a consistent display of cellular pathology. However, the translatability of murine models is limited by disparities in anatomy, body size, life span and inconsistent subtle behavior deficits that can be difficult to detect in CLN3 mutant mouse models, thereby limiting their use in preclinical studies. Here, we present a longitudinal characterization of a novel miniswine model of CLN3 disease that recapitulates the most common human pathogenic variant, an exon 7-8 deletion (CLN3Δex7/8). Progressive pathology and neuron loss is observed in various regions of the CLN3Δex7/8 miniswine brain and retina. Additionally, mutant miniswine present with retinal degeneration and motor abnormalities, similar to deficits seen in humans diagnosed with the disease. Taken together, the CLN3Δex7/8 miniswine model shows consistent and progressive Batten disease pathology, and behavioral impairment mirroring clinical presentation, demonstrating its value in studying the role of CLN3 and safety/efficacy of novel disease-modifying therapeutics.

A Scoping Review of Risk and Protective Factors for Negative Cannabis Use Consequences.

Objective: Numerous reviews have examined risk and protective factors for alcohol-related negative consequences, but no equivalent review of risk and protective factors exists for cannabis-related negative consequences (CRNCs)-a gap filled by the present study. This scoping review examined survey-based research of risk and protective factors for CRNCs such as neglecting responsibilities, blacking out, or needing more cannabis. Methods: Three databases (PubMed, PsycINFO, and Google Scholar) were searched for peer-reviewed manuscripts published between January 1, 1990, and December 31, 2021. A qualitative synthesis was performed using the matrix method and the results were organized using the socioecological model as a framework. Results: Eighty-three studies were included in the review. There was considerable variation in measures and operationalizations of CRNCs across studies. Risk factors were identified in the intrapersonal (depression, social anxiety, PTSD, impulsivity, sensation seeking, motives, expectancies), interpersonal/community (trauma, victimization, family and peer substance use, social norms), and social/policy (education, employment, community attachment, legalization, availability of substances) domains of influence. Protective behavioral strategies were a robust protective factor for CRNCs. Males consistently reported more CRNCs than females, but there were no differences observed across race. Conclusions: Future research should identify person- and product-specific patterns of CRNCs to refine theoretical models of cannabis misuse and addiction. Public health interventions to reduce the risk of negative consequences from cannabis should consider utilizing multilevel interventions to attenuate the cumulative risk from a combination of psychological, contextual, and social influences.

A Glutamate Scavenging Protocol Combined with Deanna Protocol in SOD1-G93A Mouse Model of ALS.

Amyotrophic lateral sclerosis (ALS) is a progressive disease of neuronal degeneration in the motor cortex, brainstem, and spinal cord, resulting in impaired motor function and premature demise as a result of insufficient respiratory drive. ALS is associated with dysfunctions in neurons, neuroglia, muscle cells, energy metabolism, and glutamate balance. Currently, there is not a widely accepted, effective treatment for this condition. Prior work from our lab has demonstrated the efficacy of supplemental nutrition with the Deanna Protocol (DP). In the present study, we tested the effects of three different treatments in a mouse model of ALS. These treatments were the DP alone, a glutamate scavenging protocol (GSP) alone, and a combination of the two treatments. Outcome measures included body weight, food intake, behavioral assessments, neurological score, and lifespan. Compared to the control group, DP had a significantly slower decline in neurological score, strength, endurance, and coordination, with a trend toward increased lifespan despite a greater loss of weight. GSP had a significantly slower decline in neurological score, strength, endurance, and coordination, with a trend toward increased lifespan. DP+GSP had a significantly slower decline in neurological score with a trend toward increased lifespan, despite a greater loss of weight. While each of the treatment groups fared better than the control group, the combination of the DP+GSP was not better than either of the individual treatments. We conclude that the beneficial effects of the DP and the GSP in this ALS mouse model are distinct, and appear to offer no additional benefit when combined.

Magnetic resonance brain volumetry biomarkers of CLN2 Batten disease identified with miniswine model.

Late-infantile neuronal ceroid lipofuscinosis type 2 (CLN2) disease (Batten disease) is a rare pediatric disease, with symptom development leading to clinical diagnosis. Early diagnosis and effective tracking of disease progression are required for treatment. We hypothesize that brain volumetry is valuable in identifying CLN2 disease at an early stage and tracking disease progression in a genetically modified miniswine model. CLN2R208X/R208X miniswine and wild type controls were evaluated at 12- and 17-months of age, correlating to early and late stages of disease progression. Magnetic resonance imaging (MRI) T1- and T2-weighted data were acquired. Total intercranial, gray matter, cerebrospinal fluid, white matter, caudate, putamen, and ventricle volumes were calculated and expressed as proportions of the intracranial volume. The brain regions were compared between timepoints and cohorts using Gardner-Altman plots, mean differences, and confidence intervals. At an early stage of disease, the total intracranial volume (- 9.06 cm3), gray matter (- 4.37% 95 CI - 7.41; - 1.83), caudate (- 0.16%, 95 CI - 0.24; - 0.08) and putamen (- 0.11% 95 CI - 0.23; - 0.02) were all notably smaller in CLN2R208X/R208X miniswines versus WT, while cerebrospinal fluid was larger (+ 3.42%, 95 CI 2.54; 6.18). As the disease progressed to a later stage, the difference between the gray matter (- 8.27%, 95 CI - 10.1; - 5.56) and cerebrospinal fluid (+ 6.88%, 95 CI 4.31; 8.51) continued to become more pronounced, while others remained stable. MRI brain volumetry in this miniswine model of CLN2 disease is sensitive to early disease detection and longitudinal change monitoring, providing a valuable tool for pre-clinical treatment development and evaluation.

The Impact of Childhood Trauma on Problematic Alcohol and Drug Use Trajectories and the Moderating Role of Social Support.

Adverse childhood experiences (ACE) have a strong association with alcohol and drug use; however, more research is needed to identify protective factors for this association. The present study assesses the longitudinal impact of ACE on problematic alcohol and drug use and the potential moderating effect of perceived social support. Data (n = 1404) are from a sample of Hispanic youth surveyed in high school through young adulthood. Linear growth curve models assessed the effect of ACE and perceived social support over time on problematic alcohol and drug use. Results indicated youth with ACE (vs. those without ACE) report more problematic alcohol and drug use in adolescence and have increased rates into young adulthood. Additionally, findings suggest that social support in high school may moderate the effects of ACE on problematic use over time. Among youth with high levels of support, the association of ACE with problematic alcohol and drug use was diminished. Although ACE can have a persistent impact on problematic alcohol and drug use from adolescence into adulthood, high social support during adolescence may mitigate the negative effects of ACE, lowering early problematic alcohol and drug use, offering the potential for lasting benefits.

Correlates of COVID-19 vaccination status among college students.

Objectives: Despite the widespread availability of COVID-19 vaccines in the United States, vaccine hesitancy remains high among certain groups. This study examined the correlates of being unvaccinated among a sample of students attending a single university (N = 2900) during the spring and summer of 2021, when the campus had been closed for over a year and students were preparing to return to in-person learning. Methods: Students responded to an email invitation and completed electronic surveys. Results: In multivariable logistic regression analyses, students were more likely to be unvaccinated if they were African American, identified with any political affiliation other than Democrat, were undergraduates or international students, had not traveled outside the Los Angeles during the pandemic, and/or had previously been ill with COVID-19. Conclusion: Findings indicate that culturally resonant educational interventions, and possibly vaccine requirements, are needed to promote vaccination among university students.

Identifying Nonpatient Authors of Patient Portal Secure Messages in Oncology: A Proof-of-Concept Demonstration of Natural Language Processing Methods.

PURPOSE: Patient portal secure messages are not always authored by the patient account holder. Understanding who authored the message is particularly important in an oncology setting where symptom reporting is crucial to patient treatment. Natural language processing has the potential to detect messages not authored by the patient automatically. METHODS: Patient portal secure messages from the Memorial Sloan Kettering Cancer Center were retrieved and manually annotated as a predicted unregistered proxy (ie, not written by the patient) or a presumed patient. After randomly splitting the annotated messages into training and test sets in a 70:30 ratio, a bag-of-words approach was used to extract features and then a Least Absolute Shrinkage and Selection Operator (LASSO) model was trained and used for classification. RESULTS: Portal secure messages (n = 2,000) were randomly selected from unique patient accounts and manually annotated. We excluded 335 messages from the data set as the annotators could not determine if they were written by a patient or proxy. Using the remaining 1,665 messages, a LASSO model was developed that achieved an area under the curve of 0.932 and an area under the precision recall curve of 0.748. The sensitivity and specificity related to classifying true-positive cases (predicted unregistered proxy-authored messages) and true negatives (presumed patient-authored messages) were 0.681 and 0.960, respectively. CONCLUSION: Our work demonstrates the feasibility of using unstructured, heterogenous patient portal secure messages to determine portal secure message authorship. Identifying patient authorship in real time can improve patient portal account security and can be used to improve the quality of the information extracted from the patient portal, such as patient-reported outcomes.

A Novel Porcine Model of CLN2 Batten Disease that Recapitulates Patient Phenotypes.

CLN2 Batten disease is a lysosomal disorder in which pathogenic variants in CLN2 lead to reduced activity in the enzyme tripeptidyl peptidase 1. The disease typically manifests around 2 to 4 years of age with developmental delay, ataxia, seizures, inability to speak and walk, and fatality between 6 and 12 years of age. Multiple Cln2 mouse models exist to better understand the etiology of the disease; however, these models are unable to adequately recapitulate the disease due to differences in anatomy and physiology, limiting their utility for therapeutic testing. Here, we describe a new CLN2R208X/R208X porcine model of CLN2 disease. We present comprehensive characterization showing behavioral, pathological, and visual phenotypes that recapitulate those seen in CLN2 patients. CLN2R208X/R208X miniswine present with gait abnormalities at 6 months of age, ERG waveform declines at 6-9 months, vision loss at 11 months, cognitive declines at 12 months, seizures by 15 months, and early death at 18 months due to failure to thrive. CLN2R208X/R208X miniswine also showed classic storage material accumulation and glial activation in the brain at 6 months, and cortical atrophy at 12 months. Thus, the CLN2R208X/R208X miniswine model is a valuable resource for biomarker discovery and therapeutic development in CLN2 disease.

IL-15 blockade and rapamycin rescue multifactorial loss of factor VIII from AAV-transduced hepatocytes in hemophilia A mice.

Hepatic adeno-associated viral (AAV) gene transfer has the potential to cure the X-linked bleeding disorder hemophilia A. However, declining therapeutic coagulation factor VIII (FVIII) expression has plagued clinical trials. To assess the mechanistic underpinnings of this loss of FVIII expression, we developed a hemophilia A mouse model that shares key features observed in clinical trials. Following liver-directed AAV8 gene transfer in the presence of rapamycin, initial FVIII protein expression declines over time in the absence of antibody formation. Surprisingly, loss of FVIII protein production occurs despite persistence of transgene and mRNA, suggesting a translational shutdown rather than a loss of transduced hepatocytes. Some of the animals develop ER stress, which may be linked to hepatic inflammatory cytokine expression. FVIII protein expression is preserved by interleukin-15/interleukin-15 receptor blockade, which suppresses CD8+ T and natural killer cell responses. Interestingly, mice with initial FVIII levels >100% of normal had diminishing expression while still under immune suppression. Taken together, our findings of interanimal variability of the response, and the ability of the immune system to shut down transgene expression without utilizing cytolytic or antibody-mediated mechanisms, illustrate the challenges associated with FVIII gene transfer. Our protocols based upon cytokine blockade should help to maintain efficient FVIII expression.

Forensic cardiac device analysis at the Los Angeles County Department of the Coroner: A 20-year experience.

Cardiac implantable electronic devices (CIEDs) store information continuously; however, the log of these devices is rarely analyzed in forensic practice. We retrospectively reviewed all cases referred for CIED interrogation by the Los Angeles County Department of Medical Examiner-Coroner between 2001 and 2020. According to the Department's practice, CIED interrogation may be requested for decedents in which details or cause of death are not clear from autopsy and clinical history. The CIED analysis was considered informative for the coroner's investigation either if it detected an arrhythmia or malfunction likely related to decedent's terminal event or if it was essential to determine time of death or identity of decedent. A total of 57 CIEDs were evaluated during the 20-year period. In almost half of cases (26/57: 45.6%), device analysis was informative for coroner's investigation. Arrhythmias likely related to terminal event were commonly detected (21/57: 36.8%). Device malfunction was identified as the likely cause of death in almost 10% of decedents (5/57: 8.8%), including three cases of battery depletion (3/57: 5.3%), one case of misclassification of ventricular tachycardia as supraventricular tachycardia with failure to deliver therapy (1/57: 1.7%), and one case of lead failure due to a broken pacing wire (1/57: 1.7%). Not infrequently, CIED interrogation was essential for determination of time of death (9/57: 15.8%), and there was one case (1/57: 1.7%) in which interrogation was essential for identifying the decedent. Our study shows that postmortem CIED interrogation can provide unique information regarding mechanism and time of death, and decedent's identity.