Emerging and traditional 5-α reductase inhibitors and androgen receptor antagonists for male androgenetic alopecia.

INTRODUCTION: Androgenetic alopecia (AGA) is the most prevalent cause of male hair loss, often requiring medical and/or surgical intervention. The US FDA has approved topical minoxidil and oral finasteride for male AGA treatment. However, some AGA patients fail to respond satisfactorily to these FDA-approved treatments and/or may experience side effects, based on their individual profiles. To mitigate the shortcomings of these treatments, researchers are now exploring alternative treatments such as newer 5-α reductase inhibitors (5-ARIs) and androgen receptor antagonists (ARAs). AREAS COVERED: This article reviews the safety and effectiveness of well-known 5-α reductase inhibitors (5-ARIs) like finasteride and dutasteride, as well as the newer 5-ARIs, emerging androgen receptor antagonists (ARAs), and natural products such as saw palmetto and pumpkin seed oil in the treatment of male AGA. EXPERT OPINION: Although several newer 5-ARIs, ARAs, and natural products have exhibited promise in clinical trials, additional research is essential to confirm their safety and efficacy in treating male AGA. Until additional evidence is available for these agents, the preferred treatment choices for male AGA are the FDA-approved treatments, topical minoxidil, and oral finasteride.

Chromatin accessibility profiling reveals that human fibroblasts respond to mechanical stimulation in a cell-specific manner.

Fibroblasts in the skin are highly heterogeneous, both in vivo and in vitro. One difference between follicular (dermal papilla fibroblasts [DP]) and interfollicular fibroblasts (papillary fibroblasts [PFi]) in vitro is their ability to differentiate in response to osteogenic media (OM), or mechanical stimulation. Here, we asked whether differences in the ability of DP and PFi to respond to differentiation stimuli are due to differences in chromatin accessibility. We performed chromatin accessibility and transcriptional profiling of DP and PFi in human skin, which arise from a common progenitor during development, yet display distinct characteristics in adult tissue and in vitro. We found that cells cultured in growth media had unique chromatin accessibility profiles; however, these profiles control similar functional networks. Upon introduction of a chemical perturbation (OM) to promote differentiation, we observed a divergence not only in the accessible chromatin signatures but also in the functional networks controlled by these signatures. The biggest divergence between DP and PFi was observed when we applied 2 perturbations to cells: growth in OM and mechanical stimulation (a shock wave [OMSW]). DP readily differentiate into bone in OMSW conditions, while PFi lack differentiation capability in vitro. In the DP we found a number of uniquely accessible promoters that controlled osteogenic interaction networks associated with bone and differentiation functions. Using ATAC-seq and RNA-seq we found that the combination of 2 stimuli (OMSW) could result in significant changes in chromatin accessibility associated with osteogenic differentiation, but only within the DP (capable of osteogenic differentiation). De novo motif analysis identified enrichment of motifs bound by the TEA domain (TEAD) family of transcription factors, and inter-cell comparisons (UpSet analysis) displayed large groups of genes to be unique to single cell types and conditions. Our results suggest that these 2 stimuli (OMSW) elicit cell-specific responses by modifying chromatin accessibility of osteogenic-related gene promoters.

Intratumoral T-cell receptor repertoire composition predicts overall survival in patients with pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal malignancy that is refractory to immune checkpoint inhibitor therapy. However, intratumoral T-cell infiltration correlates with improved overall survival (OS). Herein, we characterized the diversity and antigen specificity of the PDAC T-cell receptor (TCR) repertoire to identify novel immune-relevant biomarkers. Demographic, clinical, and TCR-beta sequencing data were collated from 353 patients across three cohorts that underwent surgical resection for PDAC. TCR diversity was calculated using Shannon Wiener index, Inverse Simpson index, and "True entropy." Patients were clustered by shared repertoire specificity. TCRs predictive of OS were identified and their associated transcriptional states were characterized by single-cell RNAseq. In multivariate Cox regression models controlling for relevant covariates, high intratumoral TCR diversity predicted OS across multiple cohorts. Conversely, in peripheral blood, high abundance of T-cells, but not high diversity, predicted OS. Clustering patients based on TCR specificity revealed a subset of TCRs that predicts OS. Interestingly, these TCR sequences were more likely to encode CD8+ effector memory and CD4+ T-regulatory (Tregs) T-cells, all with the capacity to recognize beta islet-derived autoantigens. As opposed to T-cell abundance, intratumoral TCR diversity was predictive of OS in multiple PDAC cohorts, and a subset of TCRs enriched in high-diversity patients independently correlated with OS. These findings emphasize the importance of evaluating peripheral and intratumoral TCR repertoires as distinct and relevant biomarkers in PDAC.

Association between alcohol use and frailty among elder adults in three Chinese cities.

BACKGROUND: Frailty is often described as a condition of the elderly and alcohol use is associated with frailty. The aim of this study is to examine the associations between alcohol use and frailty in three cities in elder adults. METHODS: A cross-sectional study was conducted in three cities in China from June 2017 to October 2018. In total, 2888 residents aged ≥65 years old were selected by using a multi-level stage sampling procedure. Alcohol use was assessed by Focusing on Cutting down, Annoyance by criticism, Guilty feeling, and Eye-openers (CAGE) four-item questionnaire. Frailty was measured by a validated Chinese version of the Fatigue, Resistance, Ambulation, Illness, and Loss of weight (FRAIL) scale. Multinomial logistic regressions were used to examine the association of alcohol use with pre-frailty and frailty after controlling for varied covariates. RESULTS: In general, the prevalence of pre-frailty and frailty was 38.64 and 20.26%, respectively. After controlling for covariates and interaction of age and problematic drinking, non-problematic drinkers neither had association with pre-frailty (OR: 1.15, 95%CI:0.86-1.52) nor with frailty (OR:0.90, 95%CI:0.60-1.36), and problematic drinkers neither had association with frailty (OR: 1.21, 95%CI:0.83-1.76), while problematic drinkers had high odd ratios of frailty (OR:3.28, 95%CI:2.02-5.33) compared with zero-drinker. CONCLUSIONS: Our study found a positive association between problematic drinking and frailty, no relationship between non-problematic drinking and (pre-)frailty compared with zero-drinking among Chinese elder adults. Based on previous findings and ours, we conclude it is important for the prevention of frailty to advocate no problematic drinking among elder adults.

Identifying indicators for the evaluation of community interventions and initiatives in a multi-site public health programme.

INTRODUCTION: Measuring success of community-level programmes and interventions is important, and indicators can provide valuable information to achieve this. However, identifying appropriate indicators can be challenging. Indicators can be identified by official local stakeholders such as local authorities, but involving communities can add value and trust to the project, with community involvement likely to improve programme sustainability. METHODS: As part of the evaluation of multi-site community initiatives, we used local health profiles to identify core indicators that overlapped sites. In addition, we engaged with members of the community during a pilot data collection training day to identify issues they identified as important for measuring health and well-being locally. RESULTS: A total of 313 indicators were identified from local profiles, with 31 indicators meeting inclusion criteria. The community identified 26 issues, collated into eight categories, only three of which were identified in core indicators. Tools were sourced or created for the other community-identified categories. DISCUSSION: The methodology identified validated indicators comparable across all sites, based on local health profiles. It also identified tools for measuring issues identified by members of the community. The exercise demonstrated disconnect between priorities of official bodies, researchers and communities, indicating multiple approaches should be considered when evaluating community initiatives.

An evaluation of the Birchwood Junk Food Café, Skelmersdale.

Food waste is an issue of global concern requiring worldwide action. In the UK, £19 billion worth of food is wasted every year. A variety of initiatives have been developed to redistribute surplus food to those in need. The Birchwood Junk Food Café in Skelmersdale combines the reduction of food waste with community and societal benefits. The University of Manchester and the Birchwood Centre conducted an evaluation of the café including a customer satisfaction survey, a long-form health and wellbeing survey and qualitative interviews. Each day the café produces a three-course menu for the public on a 'pay-as-you-feel' basis. During an 18-month period, the café intercepted 32 729 kg of food that would otherwise have gone to waste, served over 1500 people, with 3500 covers, 60 different dishes and 1200 volunteer hours. Customer satisfaction was extremely high with 88% being repeated visitors and 86% rating the café as excellent. Volunteers include youth from the local Birchwood Centre, who gain valuable experiences. Customers benefit from social interactions and additional community cohesion. The café offers an unique opportunity to impact on the wider community and provides support and structure for the volunteers.

The impact of education level on Knowledge, Attitudes and Practices towards COVID-19 in Edo State, Nigeria.

BACKGROUND: For effective Public Health measures, factors that influence Knowledge, Attitudes and Practices (KAP) need to be understood. In this paper, we document the relationship between levels of education on the KAP towards COVID-19 among the population of Edo State, Nigeria. METHODS: A cross-sectional KAP questionnaire was delivered across 13 communities. The study population was dichotomized into those who had or had not completed secondary education. Chi-square tests were conducted to determine statistical significance. RESULTS: Of the 538 participants, 30% had completed secondary education. Those with secondary education were significantly more likely to recognize symptoms of COVID-19 (P < 0.001). For attitudes, only one option showed a statistically significant association with 78.4% of those who did not complete secondary education having a high confidence in the church compared with 66% of those who had (P = 0.022). For practices, those who have completed secondary education practiced all of the supplied measures more than those who had not, except for taking herbal supplements. DISCUSSION: Higher levels of education impacts on knowledge and practices but has less impact on attitudes regarding how well certain individuals/organizations can handle the pandemic. Findings highlighted the importance of tailoring public health communication and strategies to local populations to improve the efficacy.

Mechanical stimulation of human hair follicle outer root sheath cultures activates adjacent sensory neurons.

Mechanical stimuli, such as stroking or pressing on the skin, activate mechanoreceptors transmitting information to the sensory nervous system and brain. It is well accepted that deflection of the hair fiber that occurs with a light breeze or touch directly activates the sensory neurons surrounding the hair follicle, facilitating transmission of mechanical information. Here, we hypothesized that hair follicle outer root sheath cells act as transducers of mechanical stimuli to sensory neurons surrounding the hair follicle. Using electrochemical analysis on human hair follicle preparations in vitro, we were able to show that outer root sheath cells release ATP and the neurotransmitters serotonin and histamine in response to mechanical stimulation. Using calcium imaging combined with pharmacology in a coculture of outer root sheath cells with sensory neurons, we found that the release of these three molecules from hair follicle cells leads to activation of sensory neurons.

Improved nursing home end-stage renal disease patient participation in physical therapy with onsite, more frequent dialysis.

INTRODUCTION: For end-stage renal disease (ESRD) patients residing in skilled nursing facilities (SNFs), the logistics and physical exhaustion of life-saving hemodialysis therapy often conflict with rehabilitation goals. Integration of dialysis care with rehabilitation programs in a scalable and cost-efficient manner has been a significant challenge. SNF-resident ESRD patients receiving onsite, more frequent hemodialysis (MFD) have reported rapid post-dialysis recovery. We examined whether such patients have improved Physical Therapy (PT) participation. METHODS: We conducted a retrospective electronic medical records review of SNF-resident PT participation rates within a multistate provider of SNF rehabilitation care from January 1, 2022 to June 1, 2022. We compared three groups: ESRD patients receiving onsite MFD (Onsite-MFD), ESRD patients receiving offsite, conventional 3×/week dialysis (Offsite-Conventional-HD), and the general non-ESRD SNF rehabilitation population (Non-ESRD). We evaluated physical therapy participation rates based on a predefined metric of missed or shortened (<15 min) therapy days. Baseline demographics and functional status were assessed. FINDINGS: Ninety-two Onsite-MFD had 2084 PT sessions scheduled, 12,916 Non-ESRD had 225,496 PT sessions scheduled, and 562 Offsite-Conventional-HD had 9082 PT sessions scheduled. In mixed model logistic regression, Onsite-MFD achieved higher PT participation rates than Offsite-Conventional-HD (odds ratio: 1.8, CI: 1.1-3.0; p < 0.03), and Onsite-MFD achieved equivalent PT participation rates to Non-ESRD (odds ratio: 1.2, CI: 0.3-1.9; p < 0.46). Baseline mean ± SD Charlson Comorbidity score was significantly higher in Onsite-MFD (4.9 ± 2.0) and Offsite-Conventional-HD (4.9 ± 1.8) versus Non-ESRD (2.6 ± 2.0; p < 0.001). Baseline mean self-care and mobility scores were significantly lower in Onsite-MFD versus Non-ESRD or Offsite-Conventional-HD. DISCUSSION: SNF-resident ESRD patients receiving MFD colocated with rehabilitation had higher PT participation rates than those conventionally dialyzed offsite and equivalent PT participation rates to the non-ESRD SNF-rehabilitation general population, despite being sicker, less independent, and less mobile. We report a scalable program integrating dialysis and rehabilitation care as a potential solution for ESRD patients recovering from acute hospitalization.

Comparison of oral minoxidil, finasteride, and dutasteride for treating androgenetic alopecia.

BACKGROUND: Androgenetic alopecia (AGA) is the most common cause of hair loss, often challenging to treat. While oral finasteride (1 mg/d) is an FDA-approved treatment for male AGA, oral minoxidil and oral dutasteride are not approved yet. However, clinicians have been increasingly using these two drugs off-label for hair loss. Recently, Japan and South Korea have approved oral dutasteride (0.5 mg/d) for male AGA. EFFICACY AND SAFETY: A probable efficacy ranking, in decreasing order, is - dutasteride 0.5 mg/d, finasteride 5 mg/d, minoxidil 5 mg/d, finasteride 1 mg/d, followed by minoxidil 0.25 mg/d. Oral minoxidil predominantly causes hypertrichosis and cardiovascular system (CVS) symptoms/signs in a dose-dependent manner, whereas oral finasteride and dutasteride are associated with sexual dysfunction and neuropsychiatric side effects. PHARMACOKINETICS AND PHARMACODYNAMICS: The average plasma half-lives of minoxidil, finasteride, and dutasteride are ∼4 h, ∼4.5 h, and ∼5 weeks, respectively. Minoxidil acts through multiple pathways to promote hair growth. It has been shown as a vasodilator, an anti-inflammatory agent, a Wnt/ß-catenin signaling inducer, and an antiandrogen. Finasteride inhibits 5α-reductase (5AR) type II isoenzyme, while dutasteride inhibits both type I and type II. Thus, dutasteride suppresses DHT levels more than finasteride in the serum and scalp.

Qualitative imaging features of pancreatic neuroendocrine neoplasms predict histopathologic characteristics including tumor grade and patient outcome.

OBJECTIVES: To identify PanNEN imaging features associated with tumor grade and aggressive histopathological features. METHODS: Associations between histopathological and imaging features of resected PanNEN were retrospectively tested. Histopathologic features included WHO grade, lymphovascular invasion (LVI), growth pattern (infiltrative, circumscribed), and intratumoral fibrosis (mature, immature). Imaging features included size, degree/uniformity of enhancement, progressive enhancement, contour, infiltrative appearance (infiltrativeim), calcifications, cystic components, tumor thrombus, vascular occlusion (VO), duct dilatation, and atrophy. Multinomial logistic regression analyses evaluated the magnitude of associations. Association of variables with outcome was assessed using Cox-proportional hazards regression. RESULTS: 133 patients were included. 3 imaging features (infiltrativeim, ill-defined contour [contourill], and VO) were associated with all histopathologic parameters and poor outcome. Increase in grade increased odds of contourill by 15.6 times (p = 0.0001, 95% CI 3.8-64.4). PanNEN with VO were 51.1 times (p = 0.0002, 6.5-398.6) more likely to demonstrate LVI. For PanNEN with contourill, infiltrative growth pattern was 51.3 times (p < 0.0001, 9.1-288.4), and fibrosis was 14 times (p = 0.0065, 2.1-93.7) more likely. Contourill was associated with decreased recurrence-free survival (p = 0.0003, HR 18.29, 3.83-87.3) and VO (p = 0.0004, HR6.08, 2.22-16.68) with decreased overall survival. CONCLUSIONS: Infiltrativeim, contourill, and VO on imaging are associated with higher grade/histopathological parameters linked to tumor aggression, and poor outcome.

Human hair follicles operate an internal Cori cycle and modulate their growth via glycogen phosphorylase.

Hair follicles (HFs) are unique, multi-compartment, mini-organs that cycle through phases of active hair growth and pigmentation (anagen), apoptosis-driven regression (catagen) and relative quiescence (telogen). Anagen HFs have high demands for energy and biosynthesis precursors mainly fulfilled by aerobic glycolysis. Histochemistry reports the outer root sheath (ORS) contains high levels of glycogen. To investigate a functional role for glycogen in the HF we quantified glycogen by Periodic-Acid Schiff (PAS) histomorphometry and colorimetric quantitative assay showing ORS of anagen VI HFs contained high levels of glycogen that decreased in catagen. qPCR and immunofluorescence microscopy showed the ORS expressed all enzymes for glycogen synthesis and metabolism. Using human ORS keratinocytes (ORS-KC) and ex vivo human HF organ culture we showed active glycogen metabolism by nutrient starvation and use of a specific glycogen phosphorylase (PYGL) inhibitor. Glycogen in ORS-KC was significantly increased by incubation with lactate demonstrating a functional Cori cycle. Inhibition of PYGL significantly stimulated the ex vivo growth of HFs and delayed onset of catagen. This study defines translationally relevant and therapeutically targetable new features of HF metabolism showing that human scalp HFs operate an internal Cori cycle, synthesize glycogen in the presence of lactate and modulate their growth via PYGL activity.

Longitudinal association between physical activity and health-related quality of life among community-dwelling older adults: a longitudinal study of Urban Health Centres Europe (UHCE).

BACKGROUND: Physical activity (PA) may play a key role in healthy aging and thus in promoting health-related quality of life (HRQoL). However, longitudinal studies on the association between PA and HRQoL are still scarce and have shown inconsistent results. In this study, we aimed to examine the longitudinal association between frequency of moderate PA and physical and mental HRQoL. Secondly, to assess the association between a 12-month change in frequency of moderate PA and HRQoL. METHODS: A 12-month longitudinal study was conducted in Spain, Greece, Croatia, the Netherlands, and the United Kingdom with 1614 participants (61.0% female; mean age = 79.8; SD = 5.2) included in the analyses. Two categories of the self-reported frequency of moderate PA including 1) 'regular frequency' and 2) 'low frequency' were classified, and four categories of the change in frequency of moderate PA between baseline and follow-up including 1) 'continued regular frequency', 2) 'decreased frequency', 3) 'continued low frequency' and 4) 'increased frequency' were identified. Physical and mental HRQoL were assessed by the 12-Item Short-Form Health Survey (SF-12). RESULTS: The frequency of moderate PA at baseline was positively associated with HRQoL at follow-up. Participants with a continued regular frequency had the highest HRQoL at baseline and follow-up. Participants who increased the frequency of moderate PA from low to regular had better physical and mental HRQoL at follow-up than themselves at baseline. After controlling for baseline HRQoL and covariates, compared with participants who continued a regular frequency, participants who decreased their frequency had significantly lower physical (B = -4.42; P < .001) and mental (B = -3.95; P < .001) HRQoL at follow-up; participants who continued a low frequency also had significantly lower physical (B = -5.45; P < .001) and mental (B = -4.10; P < .001) HRQoL at follow-up. The follow-up HRQoL of participants who increased their frequency was similar to those who continued a regular frequency. CONCLUSIONS: Maintaining or increasing to a regular frequency of PA are associated with maintaining or improving physical and mental HRQoL. Our findings support the development of health promotion and long-term care strategies to encourage older adults to maintain a regular frequency of PA to promote their HRQoL.

Toys or Tools? Utilization of Unmanned Aerial Systems in Mosquito and Vector Control Programs.

Organized mosquito control programs (MCP) in the United States have been protecting public health since the early 1900s. These programs utilize integrated mosquito management for surveillance and control measures to enhance quality of life and protect the public from mosquito-borne diseases. Because much of the equipment and insecticides are developed for agriculture, MCP are left to innovate and adapt what is available to accomplish their core missions. Unmanned aerial systems (UAS) are one such innovation that are quickly being adopted by MCP. The advantages of UAS are no longer conjectural. In addition to locating mosquito larval habitats, UAS affords MCP real-time imagery, improved accuracy of aerial insecticide applications, mosquito larval detection and sampling. UAS are also leveraged for applying larvicides to water in habitats that range in size from multi-acre wetlands to small containers in urban settings. Employing UAS can reduce staff exposure to hazards and the impact associated with the use of heavy equipment in sensitive habitats. UAS are utilized by MCP nationally and their use will continue to increase as technology advances and regulations change. Current impediments include a dearth of major UAS manufacturers of equipment that is tailor-made for mosquito control, pesticides that are optimized for application via UAS and regulations that limit the access of UAS to national airspace. This manuscript highlights the strengths and weaknesses of UAS within MCP, provides an update on systems and methods used, and charts the future direction of UAS technology within MCP tasked with public health protection.

Socio-demographic characteristics associated with emotional and social loneliness among older adults.

BACKGROUND: International studies provide an overview of socio-demographic characteristics associated with loneliness among older adults, but few studies distinguished between emotional and social loneliness. This study examined socio-demographic characteristics associated with emotional and social loneliness. METHODS: Data of 2251 community-dwelling older adults, included at the baseline measure of the Urban Health Centers Europe (UHCE) project, were analysed. Loneliness was measured with the 6-item De Jong-Gierveld Loneliness Scale. Multivariable logistic regression models were used to evaluate associations between age, sex, living situation, educational level, migration background, and loneliness. RESULTS: The mean age of participants was 79.7 years (SD = 5.6 years); 60.4% women. Emotional and social loneliness were reported by 29.2 and 26.7% of the participants; 13.6% experienced emotional and social loneliness simultaneously. Older age (OR: 1.16, 95% CI: 1.06-1.28), living without a partner (2.16, 95% CI: 1.73-2.70), and having a low educational level (OR: 1.82, 95% CI: 1.21-2.73), were associated with increased emotional loneliness. Women living with a partner were more prone to emotional loneliness than men living with a partner (OR: 1.78, 95% CI: 1.31-2.40). Older age (OR: 1.11, 95% CI: 1.00-1.22) and having a low educational level (OR: 1.77, 95% CI: 1.14-2.74) were associated with increased social loneliness. Men living without a partner were more prone to social loneliness than men living with a partner (OR: 1.94, 95% CI: 1.35-2.78). CONCLUSIONS: Socio-demographic characteristics associated with emotional and social loneliness differed regarding sex and living situation. Researchers, policy makers, and healthcare professionals should be aware that emotional and social loneliness may affect older adults with different socio-demographic characteristics.

Less is more in the difficult gallbladder: recent evolution of subtotal cholecystectomy in a single HPB unit.

INTRODUCTION: Subtotal cholecystectomy (SC) is a technique to manage the difficult gallbladder and avoid hazardous dissection and biliary injury. Until recently it was used infrequently. However, because of reduced exposure to open total cholecystectomy in resident training, we recently adopted subtotal cholecystectomy as the bail-out procedure of choice for resident teaching. This study reports our experience and outcomes with subtotal cholecystectomy in the years immediately preceding adoption and since adoption. METHODS: A retrospective analysis was conducted of patients undergoing SC from July 2010 to June 2019. Outcomes, including bile leak, reoperation and need for additional procedures, were analyzed. Complications were graded by the Modified Accordion Grading Scale (MAGS). RESULTS: 1571 cholecystectomies were performed of which 71 were SC. Subtotal cholecystectomy patients had several indicators of difficulty including prior attempted cholecystectomy and previous cholecystostomy tube insertion. The most common indication for SC was marked inflammation in the hepatocystic triangle (51%). As our experience increased, fewer patients required open conversion to accomplish SC and SC was completed laparoscopically, usually subtotal fenestrating cholecystectomy (SFC). Most patients (85%) had a drain placed and 28% were discharged with a drain. The highest MAGS complication observed was grade 3 (11 patients, 15%). Six patients had a bile leak from the cystic duct resolved by ERCP. At mean follow-up of about 1 year no patient returned with recurrent symptoms. CONCLUSIONS: Subtotal fenestrating cholecystectomy is a useful technique to avoid biliary injury in the difficult gallbladder and can be performed with very satisfactory rates of bile fistula, ERCP, and reoperation.

Strong population differentiation in lingcod (Ophiodon elongatus) is driven by a small portion of the genome.

Delimiting intraspecific genetic variation in harvested species is crucial to the assessment of population status for natural resource management and conservation purposes. Here, we evaluated genetic population structure in lingcod (Ophiodon elongatus), a commercially and recreationally important fishery species along the west coast of North America. We used 16,749 restriction site-associated DNA sequencing (RADseq) markers, in 611 individuals collected from across the bulk of the species range from Southeast Alaska to Baja California, Mexico. In contrast to previous population genetic work on this species, we found strong evidence for two distinct genetic clusters. These groups separated latitudinally with a break near Point Reyes off Northern California, and there was a high frequency of admixed individuals in close proximity to the break. F-statistics corroborate this genetic break between northern and southern sampling sites, although most loci are characterized by low FST values, suggesting high gene flow throughout most of the genome. Outlier analyses identified 182 loci putatively under divergent selection, most of which mapped to a single genomic region. When individuals were grouped by cluster assignment (northern, southern, and admixed), 71 loci were fixed between the northern and southern cluster, all of which were identified in the outlier scans. All individuals identified as admixed exhibited near 50:50 assignment to northern and southern clusters and were heterozygous for most fixed loci. Alignments of RADseq loci to a draft lingcod genome assembly and three other teleost genomes with chromosome-level assemblies suggest that outlier and fixed loci are concentrated on a single chromosome. Similar genomic patterns have been attributed to chromosomal inversions in diverse taxonomic groups. Regardless of the evolutionary mechanism, these results represent novel observations of genetic structure in lingcod and designate clear evolutionary units that could be used to inform fisheries management.

ELX-02 Generates Protein via Premature Stop Codon Read-Through without Inducing Native Stop Codon Read-Through Proteins.

ELX-02 is a clinical stage, small-molecule eukaryotic ribosomal selective glycoside acting to induce read-through of premature stop codons (PSCs) that results in translation of full-length protein. However, improved read-through at PSCs has raised the question of whether native stop codon (NSC) fidelity would be impacted. Here, we compare read-through by ELX-02 in PSC and NSC contexts. DMS-114 cells containing a PSC in the TP53 gene were treated with ELX-02 and tested for increased nuclear p53 protein expression while also monitoring two other proteins for NSC read-through. Additionally, blood samples were taken from healthy subjects pre- and post-treatment with ELX-02 (0.3-7.5 mg/kg). These samples were processed to collect white blood cells and then analyzed by western blot to identify native and potentially elongated proteins from NSC read-through. In a separate experiment, lymphocytes cultivated with vehicle or ELX-02 (20 and 100 µg/ml) were subjected to proteomic analysis. We found that ELX-02 produced significant read-through of the PSC found in TP53 mRNA in DMS-114 cells, resulting in increased p53 protein expression and consistent with decreased nonsense-mediated mRNA degradation. NSC read-through protein products were not observed in either DMS-114 cells or in clinical samples from subjects dosed with ELX-02. The number of read-through proteins identified by using proteomic analysis was lower than estimated, and none of the NSC read-through products identified with >2 peptides showed dose-dependent responses to ELX-02. Our results demonstrate significant PSC read-through by ELX-02 with maintained NSC fidelity, thus supporting the therapeutic utility of ELX-02 in diseases resulting from nonsense alleles. SIGNIFICANCE STATEMENT: ELX-02 produces significant read-through of premature stop codons leading to full-length functional protein, demonstrated here by using the R213X mutation in the TP53 gene of DMS-114 cells. In addition, three complementary techniques suggest that ELX-02 does not promote read-through of native stop codons at concentrations that lead to premature stop codon read-through. Thus, ELX-02 may be a potential therapeutic option for nonsense mutation-mediated genetic diseases.

Accuracy of Grading in Pancreatic Neuroendocrine Neoplasms and Effect on Survival Estimates: An Institutional Experience.

BACKGROUND: Accurate grading of neuroendocrine neoplasms (NENs) is crucial for proper assessment of prognosis. Estimation of the proliferative indices, if not performed properly, is largely erroneous due to significant intratumoral heterogeneity. We sought to establish the degree of error in the grading of a cohort of curatively resected pancreatic NENs (PanNENs) and the theoretical impact of that in a larger cohort of Surveillance, Epidemiology, and End Results (SEER) patients. METHODS: A retrospective query of an institutional surgical database was performed from 2000 to 2018 to identify optimally resected PanNENs, which were reviewed by two gastrointestinal pathologists and regraded according to the WHO 2017 classification. Overall survival and recurrence-free survival were estimated using the Kaplan-Meier method for original and new grading systems, respectively and Cox proportional hazards models were used to evaluate the effect of the interested variables, including new grading systems. RESULTS: A total of 176 cases were identified. After regrading, 17/64 (26.6%) G1 neoplasms were classified as G2 and 12/95 (12.6%) G2 neoplasms were classified as G1, while 1/11 (9.1%) G3 neoplasms were classified as G2. Our expert gastrointestinal pathologists agreed on 97% of reclassified cases by blind review. Application of the G1/G2 misclassification errors on various groups, including PanNENs, in a SEER database of 1385 patients rendered the reported survival differences nonsignificant (1000 repetitions; p = 0.063, 95% confidence interval 0.056-0.070). CONCLUSIONS: Mischaracterization of grade is common in optimally resected PanNENs but is eliminated with proper training and adherence to guidelines. The discrepancy rates can cast doubt on the generally accepted survival differences between G1 and G2 patients, as surmised by large database analyses.