Behavioral and transcriptomic analyses of mecp2 function in zebrafish.

Rett syndrome (RTT), a human neurodevelopmental disorder characterized by severe cognitive and motor impairments, is caused by dysfunction of the conserved transcriptional regulator Methyl-CpG-binding protein 2 (MECP2). Genetic analyses in mouse Mecp2 mutants, which exhibit key features of human RTT, have been essential for deciphering the mechanisms of MeCP2 function; nonetheless, our understanding of these complex mechanisms is incomplete. Zebrafish mecp2 mutants exhibit mild behavioral deficits but have not been analyzed in depth. Here, we combine transcriptomic and behavioral assays to assess baseline and stimulus-evoked motor responses and sensory filtering in zebrafish mecp2 mutants from 5 to 7 days post-fertilization (dpf). We show that zebrafish mecp2 function is required for normal thigmotaxis but is dispensable for gross movement, acoustic startle response, and sensory filtering (habituation and sensorimotor gating), and reveal a previously unknown role for mecp2 in behavioral responses to visual stimuli. RNA-seq analysis identified a large gene set that requires mecp2 function for correct transcription at 4 dpf, and pathway analysis revealed several pathways that require MeCP2 function in both zebrafish and mammals. These findings show that MeCP2's function as a transcriptional regulator is conserved across vertebrates and supports using zebrafish to complement mouse modeling in elucidating these conserved mechanisms.

Determining the MMPI-3 SUI scale's cross-sectional and prospective utility in suicide risk assessment.

OBJECTIVE: In-depth suicide risk assessments are particularly important to long-term suicide prevention. Broadband measures of psychopathology, such as the Minnesota Multiphasic Personality Inventory (MMPI) instruments, assess suicide risk factors and various mental health comorbidities. With the recent release of the MMPI-3, the Suicidal/Death Ideation (SUI) scale underwent revisions to improve its construct validity and detection of suicide risk factors. Thus, we hypothesized the MMPI-3 SUI scale would demonstrate medium to large associations with suicidal experience and behaviors, future ideation, and interpersonal risk factors of suicide. METHODS: A sample of 124 college students screened for elevated depressive symptoms completed a brief longitudinal study. Participants completed a baseline session including the MMPI-3 and criterion measures and three brief follow-ups every 2 weeks. RESULTS: SUI scores were most robustly associated with increased risk for past suicidal ideation, planning, and perceived burdensomeness. Prospectively assessed suicidal ideation was also meaningfully associated with SUI. SUI scale elevations indicate an increased risk of suicide-related risk factors. CONCLUSION: The MMPI-3 is a valuable tool to inform long-term suicide prevention for those experiencing elevated depressive symptoms as the SUI scale can assess past, current, and future suicide-related risk factors, including suicidal ideation and behaviors.

When Minutes Matter: A Comparison of Whole Blood Collection Techniques.

BACKGROUND: Fast and reliable blood collection is critical to emergency walking blood banks (WBB) because mortality significantly declines when blood is quickly administered to a warfighter with hemorrhagic shock. Phlebotomy for WBB is accomplished via either the "straight stick" (SS) or "ruggedized lock" (RL) method. SS comprises a 16-gauge phlebotomy needle connected to a blood collection bag via tubing. The RL device collects blood through the same apparatus, but has a capped, intravenous (IV) catheter between the needle and the donor's arm. This is the first study to compare these two methods in battlefield-relevant metrics. METHODS: Military first responders and licensed medical providers (N=86) were trained in SS and RL as part of fresh whole blood training exercises. Outcomes included venipuncture success rates, time to IV access, blood collection times, total time, and user preferences, using a within-subjects crossover design. Data were analyzed using ANOVA and nonparametric statistics at p<0.05. RESULTS: SS outperformed RL in first venipuncture success rates (76% vs. 64%, p=0.07), IV access times (448 [standard error of the mean; SE 23] vs. 558 [SE 31] s, p<0.01), and blood collection bag fill times (573 [SE 48] vs. 703 [SE 44] s, p<0.05), resulting in an approximate 3.5-minute faster time overall. Survey data were mixed, with users perceiving SS as simpler and faster, but RL as more reliable and secure. CONCLUSION: SS is optimal when timely collection is imperative, while RL may be preferable when device stability or replacing the collection bag is a consideration.

Assessment of PTSD and Trauma Symptoms With the MMPI-3 in College Students: Validity and Incremental Utility of the Anxiety Related Experiences (ARX) Scale.

Post Traumatic Stress Disorder (PTSD) is heterogeneous in nature, which complicates diagnostic efforts and makes accurate assessment tools critical. The MMPI family of instruments are widely used broadband measures of psychopathology, including trauma symptomology. The MMPI-3's Anxiety Related Experiences scale (ARX) is an expansion of the MMPI-2-RF Anxiety (AXY) scale which has historically represented the MMPI family's best measure of trauma symptoms. This study expands research on ARX in 2 samples of college students (n = 332 [PCL-5 Criterion] & n = 58 [CAPS-5 Criterion]) by examining ARX's incremental, criterion, and classification validity. ARX incrementally predicted PCL-5 total and cluster scores beyond that accounted for by AXY (R2Δ = .01-.09). ARX accounted for the most unique variance, beyond RCd and RC7, in CAPS-5 interview ratings of intrusion symptoms (R2Δ = .16). ARX was strongly related to trauma symptomology broadly (r = .42-.58) and demonstrated strong screening ability at T65 (sensitivity = .37-.40; specificity = .91-.92) and stronger diagnostic screening at T75 (sensitivity = .31; specificity = .93). We discuss clinical considerations when using ARX for assessing PTSD.

Feasibility Cues during a Sexual Encounter Impact the Strength of Heterosexual Women's Orgasm Goal Pursuit.

The "orgasm gap" for women who have sex with men remains a pressing gender equity issue. Recent research found that women who pursued orgasm as a goal were more likely to have one. The current research replicated this relationship between orgasm goal pursuit and orgasm frequency for heterosexual women, and found that this relationship did not exist for heterosexual men (Study 1). Then, across two experimental studies, we examined how heterosexual women vary their orgasm goal pursuit across sexual encounters. In Study 2, women who read that a hypothetical sexual encounter would be "quick" reported less intent to pursue orgasm than women who were told they could "take their time" or received no time information. In Study 3, women who read that their hypothetical sexual partner seemed selfish reported less intent to pursue orgasm than women who were given a non-selfish partner or no partner information. Importantly, these effects were mediated by women's perceived orgasm likelihood in the scenario. These results suggest that women shift their pursuit of orgasm depending on cues which signal whether orgasm will be feasible. This research used self-regulation theory to understand women's motivations for pursuing orgasm during sexual encounters with men, with implications for reducing the orgasm gap.

Investigating the Validity of the MMPI-3 Eating Concerns (EAT) scale across Men and Women in a University Sample.

Disordered eating is a major health epidemic that occurs at disproportionate rates among young adults and for which gender plays a major role in symptom presentation. Broadband psychological instruments have historically not included disordered eating as a core scale construct. The recent release of the Minnesota Multiphasic Personality Inventory-3 (MMPI-3) offers an opportunity to address this shortcoming through the newly developed Eating Concerns Scale (EAT) for which the existing literature is promising but limited. This study expands research on EAT by investigating its validity and comparing findings across gender. In 345 college students (102 men, 243 women), we examined gender differences between men and women in the EAT scale's structure, item endorsement rates, mean scores, and correlations with measures of body image and eating pathology. Differences emerged in item endorsement rate, scale score elevation rate, and correlation magnitudes. Broadly, findings further support EAT's use in detecting eating pathology and highlight ways in which the EAT scale may not effectively capture masculine expressions of eating pathology, namely binging and purging behaviors. To assess eating pathology more comprehensively, clinicians and researchers should consider including assessments of eating pathology inclusive of masculine eating patterns. Limitations and future research directions are also discussed.

Systemic Arterial Oxygen Levels Differentiate Pre- and Post-capillary Predominant Hemodynamic Abnormalities During Exercise in Undifferentiated Dyspnea on Exertion.

BACKGROUND: Whether systemic oxygen levels (SaO2) during exercise can provide a window into invasively derived exercise hemodynamic profiles in patients with undifferentiated dyspnea on exertion is unknown. METHODS: We performed cardiopulmonary exercise testing with invasive hemodynamic monitoring and arterial blood gas sampling in individuals referred for dyspnea on exertion. Receiver operator analysis was performed to distinguish heart failure with preserved ejection fraction from pulmonary arterial hypertension. RESULTS: Among 253 patients (mean ± SD, age 63 ± 14 years, 55% female, arterial O2 [PaO2] 87 ± 14 mmHg, SaO2 96% ± 4%, resting pulmonary capillary wedge pressure [PCWP] 18 ± 4mmHg, and pulmonary vascular resistance [PVR] 2.7 ± 1.2 Wood units), there was no exercise PCWP threshold, measured up to 49 mmHg, above which hypoxemia was consistently observed. Exercise PaO2 was not correlated with exercise PCWP (rho = 0.04; P = 0.51) but did relate to exercise PVR (rho = -0.46; P < 0.001). Exercise PaO2 and SaO2 levels distinguished left-heart-predominant dysfunction from pulmonary-vascular-predominant dysfunction with an area under the curve of 0.89 and 0.89, respectively. CONCLUSION: Systemic O2 levels during exercise distinguish relative pre- and post-capillary pulmonary hemodynamic abnormalities in patients with undifferentiated dyspnea. Hypoxemia during upright exercise should not be attributed to isolated elevation in left heart filling pressures and should prompt consideration of pulmonary vascular dysfunction.

Cross validation of the Personality Assessment Inventory (PAI) Cognitive Bias Scale of Scales (CB-SOS) over-reporting indicators in a military sample.

Following the development of the Cognitive Bias Scale (CBS), three other cognitive over-reporting indicators were created. This study cross-validates these new Cognitive Bias Scale of Scales (CB-SOS) measurements in a military sample and contrasts their performance to the CBS. We analyzed data from 288 active-duty soldiers who underwent neuropsychological evaluation. Groups were established based on performance validity testing (PVT) failure. Medium effects (d = .71 to .74) were observed between those passing and failing PVTs. The CB-SOS scales have high specificity (≥.90) but low sensitivity across the suggested cut scores. While all CB-SOS were able to achieve .90, lower scores were typically needed. CBS demonstrated incremental validity beyond CB-SOS-1 and CB-SOS-3; only CB-SOS-2 was incremental beyond CBS. In a military sample, the CB-SOS scales have more limited sensitivity than in its original validation, indicating an area of limited utility despite easier calculation. The CBS performs comparably, if not better, than CB-SOS scales. CB-SOS-2's differences in performance in this study and its initial validation suggest that its psychometric properties may be sample dependent. Given their ease of calculation and relatively high specificity, our study supports the interpretation of elevated CB-SOS scores indicating those who are likely to fail concurrent PVTs.

Improving maternal update rates within the first hour of NICU admission.

BACKGROUND: Timely communication is essential in attaining maternal satisfaction, developing an excellent physician-patient rapport, and increasing trust. This study reports a significant improvement in maternal communication rates through the quality improvement method. METHODS: An educational module was developed, and NICU staff was presented with the slides, followed by a performance questionnaire to demonstrate understanding. The first phase was completed by obtaining feedback from mothers through a questionnaire. The first plan-do-study-act (PDSA) cycle, carried out for eight weeks looking at the rates of the maternal update provided within an hour of admission of their neonates to the NICU, was followed by the second PDSA cycle, carried out for ten weeks. The improvement was calculated using conventional statistics and a statistical process control chart. RESULTS: During the first phase of the study, thirty-six percent of the mothers were updated within an hour of admission of their neonates to the NICU. During the first PDSA cycle, we did not notice a special cause variation or process change. A significant shift, eight consecutive points above the mean, was noted on the control chart during PDSA cycle 2. The mean±SD of the weekly update rate increased significantly during PDSA cycle 2 (76.8±11) compared to PDSA cycle 1 (47.5±14), p-value = 0.0002. CONCLUSION: We improved the maternal update rates through the educational module following the QI improvement model using the PDSA cycles.

Evaluation of a Passive Wearable Device for Post-Stroke Shoulder Abduction Support.

Post-stroke upper extremity function can be improved by devices that support shoulder abduction. However, many of these devices provide limited assistance in activities of daily living due to their complexity and encumbrance. We developed and evaluated a passive, lightweight (0.6 kg) wearable device consisting of an aluminum frame and elastic bands attached to a posture vest to aid in shoulder abduction. The number and thickness of bands can be adjusted to provide supportive forces to the affected arm. We measured reachable workspace area and Wolf Motor Function Test (WMFT) performance in people with a history of stroke (n = 11) with and without the wearable. The device increased workspace area in 6 participants and improved average WMFT functional and timing scores in 7 and 12 tasks, respectively, out of 16 total tasks. On average, participants increased their arm motion within 20 cm of shoulder level by 22.4% and decreased their hand's average distance from trunk by 15.2%, both improvements in the device case.

MiR-324-5p inhibition after intrahippocampal kainic acid-induced status epilepticus does not prevent epileptogenesis in mice.

Background: Acquired epilepsies are caused by an initial brain insult that is followed by epileptogenesis and finally the development of spontaneous recurrent seizures. The mechanisms underlying epileptogenesis are not fully understood. MicroRNAs regulate mRNA translation and stability and are frequently implicated in epilepsy. For example, antagonism of a specific microRNA, miR-324-5p, before brain insult and in a model of chronic epilepsy decreases seizure susceptibility and frequency, respectively. Here, we tested whether antagonism of miR-324-5p during epileptogenesis inhibits the development of epilepsy. Methods: We used the intrahippocampal kainic acid (IHpKa) model to initiate epileptogenesis in male wild type C57BL/6 J mice aged 6-8 weeks. Twenty-four hours after IHpKa, we administered a miR-324-5p or scrambled control antagomir intracerebroventricularly and implanted cortical surface electrodes for EEG monitoring. EEG data was collected for 28 days and analyzed for seizure frequency and duration, interictal spike activity, and EEG power. Brains were collected for histological analysis. Results: Histological analysis of brain tissue showed that IHpKa caused characteristic hippocampal damage in most mice regardless of treatment. Antagomir treatment did not affect latency to, frequency, or duration of spontaneous recurrent seizures or interictal spike activity but did alter the temporal development of frequency band-specific EEG power. Conclusion: These results suggest that miR-324-5p inhibition during epileptogenesis induced by status epilepticus does not convey anti-epileptogenic effects despite having subtle effects on EEG frequency bands. Our results highlight the importance of timing of intervention across epilepsy development and suggest that miR-324-5p may act primarily as a proconvulsant rather than a pro-epileptogenic regulator.

Accuracy of portable ultrasound for obstetric biometry.

OBJECTIVE: We assessed the accuracy of two portable ultrasound machines (PUM) in obtaining fetal biometry and estimating gestational age. METHODS: We analyzed data from the Fetal Age Machine Learning Initiative, an observational study of pregnant women in the United States and Zambia. Each participant underwent assessment by an experienced sonographer using both a high-specification ultrasound machine (HSUM) and a PUM (either Butterfly iQ or Clarius C3) to measure fetal biometry and calculate estimated gestational age (EGA) at each visit. Through comparison of paired PUM-HSUM scans, we estimated agreement between individual biometry measurements and aggregate gestational age estimates by reporting mean difference, along with intraclass correlation coefficient (ICC) and Bland-Altman plots, adjusting for trend. RESULTS: 881 participants contributed 1386 paired PUM-HSUM ultrasound studies between April and December 2021. PUM studies included 991 Butterfly and 395 Clarius. Gestational age at scan ranged from 7 to 38 weeks. Compared to HSUM, the Butterfly PUM had a mean difference of -0.20 days (95%CI±0.40) in the 1st trimester and -0.68 days (95%CI±0.68) in the 2nd/3rd trimesters. Also compared to HSUM, the Clarius PUM had a mean difference of 0.47 days (95%CI±0.64) in the 1st trimester and -1.67 days (95%CI±0.43) in the 2nd/3rd trimesters. ICCs were 0.989 or greater throughout. Increasing gestational age was associated with increasing error and absolute error. Both PUM devices demonstrated a modest trend toward underestimation of EGA at advancing gestational ages in 2nd/3rd trimester scans, compared to HSUM. CONCLUSION: Both the Butterfly iQ and Clarius C3 PUM devices were highly accurate in performing fetal biometry in a diverse population from the US and Zambia. This article is protected by copyright. All rights reserved.

Contrasting MMPI-3 validity scale effectiveness differences across in-person and telehealth administration procedures.

Psychological assessment underwent substantive challenges and changes when the COVID-19 pandemic began, and these changes are likely to endure given the rapid growth of telehealth clinical practice and assessment research using virtual procedures. COVID-19-related changes to assessment practices have impacted accordingly how we study overreporting scale functioning, including the modality through which we administer measures. No available research provides direct comparisons of overreporting scale effectiveness within simulation research across in-person and telehealth modalities, despite early support for novel instruments relying on remote procedures within the historic context of the pandemic. We used simulated feigning conditions collected using best telehealth practices to examine if, and how, overreporting scales differed in effectiveness by evaluating mean scores, elevation rates, and classification accuracy statistics, relative to parallel in-person conditions. Results indicate no meaningful differences in scale effectiveness, particularly when exclusion procedures included a posttest questionnaire. Our findings support telehealth assessment practice and the integration of research collected virtually into the traditional, in-person feigning literature. Limitations and future directions are discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Neurodevelopmental outcomes in children and adults with Fetal Valproate Spectrum Disorder: A contribution from the ConcePTION project.

AIM: To describe the neurodevelopmental phenotype of older children and adults with a diagnosis of Fetal Valproate Spectrum Disorder (FVSD). METHODS: In this cross-sectional study, 90 caregivers were recruited and completed a series of questionnaires regarding the neurodevelopmental outcomes of 146 individuals aged 7-37 years (M = 18.1), including individuals with a formal diagnosis of FVSD (n = 99), individuals exposed to Valproate but without an FVSD diagnosis (n = 24), and individuals not exposed to Valproate (N = 23). The mean dose of valproate exposure for individuals with an FVSD diagnosis was 1470 mg/day. RESULTS: Individuals with a diagnosis of FVSD showed significantly higher levels of moderate (43.4%) and severe (14.4%) cognitive impairment than other groups (p = 0.003), high levels of required formal educational support (77.6%), and poorer academic competence than individuals not exposed to Valproate (p = 0.001). Overall psychosocial problems (p = 0.02), internalising problems (p = 0.05) and attention problems (p = 0.001), but not externalising problems, were elevated in individuals with a diagnosis of FVSD. Rates of neurodevelopmental disorders, particularly autistic spectrum disorders (62.9%) and sensory problems (80.6%) are particularly central to the FVSD phenotype. There was no evidence of a statistical dose-dependent effect, possibly due to the high mean dose of exposure having a uniformly negative impact across the sample. Individuals with FVSD had required a significant number of health and child development services. INTERPRETATION: Children and young adults with a diagnosis of FVSD are at an increased risk of a range of altered neurodevelopmental outcomes, highlighting the need for a multidisciplinary approach to clinical management across the lifespan.

Zileuton ameliorates aminoglycoside and polymyxin-associated acute kidney injury in an animal model.

OBJECTIVES: Aminoglycosides and polymyxins are antibiotics with in vitro activity against MDR Gram-negative bacteria. However, their clinical use is hindered by dose-limiting nephrotoxicity. The objective of this project was to determine if zileuton can reduce nephrotoxicity associated with amikacin and polymyxin B in a rat model of acute kidney injury. METHODS: Sprague Dawley rats (n = 10, both genders) were administered either amikacin (300 mg/kg) or polymyxin B (20 mg/kg) daily for 10 days. Zileuton (4 and 10 mg/kg) was delivered intraperitoneally 15 min before antibiotic administration. Blood samples were collected at baseline and daily to determine serum creatinine concentration. Nephrotoxicity was defined as a ≥2× elevation of baseline serum creatinine. Time-to-event analysis and log rank test were used to compare the onset of nephrotoxicity in different cohorts. Histopathological analysis was also conducted to characterize the extent of kidney injury. RESULTS: Animals receiving amikacin or polymyxin B alone had nephrotoxicity rates of 90% and 100%, respectively. The overall rate was reduced to 30% in animals receiving adjuvant zileuton. The onset of nephrotoxicity associated with amikacin and polymyxin B was also significantly delayed by zileuton at 4 and 10 mg/kg, respectively. Histopathology confirmed reduced kidney injury in animals receiving amikacin concomitant with zileuton. CONCLUSIONS: Our pilot data suggest that zileuton has the potential to attenuate nephrotoxicity associated with last-line antibiotics. This would allow these antibiotics to treat MDR Gram-negative bacterial infections optimally without dose-limiting constraints. Further studies are warranted to optimize drug delivery and dosing in humans.

In vitro model to simulate multiple drugs with distinct elimination half-lives.

OBJECTIVE: The prevalence of drug resistance in pathogens such as HIV and selected bacteria has been steadily rising, resulting in an increased need for using multiple agents concurrently. Agents used in these combination therapies may have different elimination half-lives in humans. There is an unmet need for in vitro models to evaluate the efficacy of these combinations to guide early drug development. In order to realistically reflect in vivo conditions, useful in vitro model systems must be capable of simulating multiple pharmacokinetic profiles with distinct elimination half-lives. The goal of this study was to experimentally simulate four pharmacokinetic profiles with distinct elimination half-lives in an in vitro hollow-fibre system. METHODS: For illustrative purposes, fluctuating exposures of ceftriaxone were simulated with distinct half-lives of 1, 2.5, 8, and 12 hours. A parallel experimental setup was used to independently connect four supplemental reservoirs to a central reservoir. Target maximum concentration was achieved by direct drug dosing into the central reservoir; supplemental reservoirs were also dosed to offset the rapid drug elimination rate from the central reservoir. Serial pharmacokinetic samples were obtained from the central reservoir, assayed by a spectrophotometric method, and characterized by a one-compartment model. RESULTS: The observed maximum concentrations and elimination half-lives were in agreement with the expected values obtained from the mathematical predictions. CONCLUSIONS: This in vitro experimental system can be used to evaluate the efficacy of up to four-drug combinations against multidrug-resistant bacteria or HIV-infected mammalian cells. The established framework represents an adaptable tool to advance the field of combination therapy.

Preoperative investigations in thyroglossal duct cyst surgery: a 9-year experience and proposed practice guide.

INTRODUCTION: Thyroglossal duct cysts (TGDC) are the commonest midline developmental cyst and have a bimodal age distribution. They usually develop in an infrahyoid position. A National survey of TGDC practice among otolaryngologists in 2012 recommended preoperative investigation with ultrasound, with or without blood tests. METHODS: Retrospective analysis of preoperative investigation for clinically diagnosed TGDC surgeries between 2012 and 2020 in a single tertiary centre was undertaken. This was collated alongside postoperative outcomes (histology, recurrence, hypothyroidism). Comparison was made with the 2012 national survey. RESULTS: Ninety-five cases of thyroglossal duct surgery in children and adults were studied. Demographic data were comparable to the literature. Ultrasonography was the most commonly used preoperative investigation. Histology of 71% of excised cysts confirmed TGDC with another 8% reported as development cysts. The excision of the cyst with a cuff of strap muscles and middle portion of hyoid bone had the lowest recurrence rate, which was 4% overall in this study. There were no cases of ectopic thyroid tissue or postoperative hypothyroidism. DISCUSSION AND CONCLUSION: Nearly a decade of thyroglossal duct cyst excisions in a large-volume unit gave detail on actual preoperative practice and outcomes. Practice was found to be consistent overall with the 2012 recommendations although not standardised across all cases. From this experience, and a literature review, a visual guide on preoperative investigations for different age groups in the form of a flow chart is proposed to reduce risk of complications while avoiding unnecessary investigations.

Preparing Future Surgeons: An Evaluation of Academic Surgeons' Views on Laparoscopic Simulation Training for Medical Students.

Introduction With the development of laparoscopic simulation, medical students are motivated to expand their knowledge and proficiency in basic surgical skills. This study aims to demonstrate their capability and readiness for surgical clerkships and, ultimately, surgical residency. This study's primary objective is to ascertain academic surgeons' perspectives regarding laparoscopic simulation in undergraduate medical education and to determine if early exposure may afford medical students additional opportunities during their surgical clerkships. Methods A survey was created to ascertain surgeon perspectives on medical students' early exposure to laparoscopic simulation. Five-point Likert scales were used to indicate surgeon perspectives. The survey was conducted over the two days of the meeting; all attendees' meeting inclusion criteria were encouraged to participate. Surgeons practicing within the state of Alabama, with prior experience overseeing the development and training of medical students before June 1, 2022, and attending the AL Chapter American College of Surgeons 2022 Annual Meeting were eligible to complete the survey. Only completed surveys were included for analysis. Statement 1: "Pre-clinical exposure to laparoscopic simulators is beneficial to the training and development of medical students pursuing a surgical career." Statement 2: "I am more likely to allow medical students to participate in laparoscopic surgery cases if they have had prior exposure to, and training with, laparoscopic simulators." Results An on-site survey was conducted among 18 surgeons consisting of 14 full-time faculty attendings, two post-graduate year-five residents, and two post-graduate year-three residents, all practicing in academic medicine with experience overseeing the training of medical students. In response to Statement 1, 33.3% of respondents strongly agree and 66.6% agree. In response to Statement 2, 61.1% of respondents strongly agree, 33.3% agree, and 5.6% were undecided. Discussion Our study provides evidence to support the inclusion of laparoscopic simulation training in undergraduate medical education to enhance medical students' basic surgical skills and improve their clinical experience. Further research could inform the development of effective laparoscopic simulation training programs that prepare medical students transitioning to surgical residency.

A clinical-grade liquid biomarker detects neuroendocrine differentiation in prostate cancer.

BackgroundNeuroendocrine prostate cancer (NEPC) is an aggressive subtype, the presence of which changes the prognosis and management of metastatic prostate cancer.MethodsWe performed analytical validation of a Circulating Tumor Cell (CTC) multiplex RNA qPCR assay to identify the limit of quantification (LOQ) in cell lines, synthetic cDNA, and patient samples. We next profiled 116 longitudinal samples from a prospectively collected institutional cohort of 17 patients with metastatic prostate cancer (7 NEPC, 10 adenocarcinoma) as well as 265 samples from 139 patients enrolled in 3 adenocarcinoma phase II trials of androgen receptor signaling inhibitors (ARSIs). We assessed a NEPC liquid biomarker via the presence of neuroendocrine markers and the absence of androgen receptor (AR) target genes.ResultsUsing the analytical validation LOQ, liquid biomarker NEPC detection in the longitudinal cohort had a per-sample sensitivity of 51.35% and a specificity of 91.14%. However, when we incorporated the serial information from multiple liquid biopsies per patient, a unique aspect of this study, the per-patient predictions were 100% accurate, with a receiver-operating-curve (ROC) AUC of 1. In the adenocarcinoma ARSI trials, the presence of neuroendocrine markers, even while AR target gene expression was retained, was a strong negative prognostic factor.ConclusionOur analytically validated CTC biomarker can detect NEPC with high diagnostic accuracy when leveraging serial samples that are only feasible using liquid biopsies. Patients with expression of NE genes while retaining AR-target gene expression may indicate the transition to neuroendocrine differentiation, with clinical characteristics consistent with this phenotype.FundingNIH (DP2 OD030734, 1UH2CA260389, R01CA247479, and P30 CA014520), Department of Defense (PC190039 and PC200334), and Prostate Cancer Foundation (Movember Foundation - PCF Challenge Award).

Reversible epigenetic alterations regulate class I HLA loss in prostate cancer.

Downregulation of HLA class I (HLA-I) impairs immune recognition and surveillance in prostate cancer and may underlie the ineffectiveness of checkpoint blockade. However, the molecular mechanisms regulating HLA-I loss in prostate cancer have not been fully explored. Here, we conducted a comprehensive analysis of HLA-I genomic, epigenomic and gene expression alterations in primary and metastatic human prostate cancer. Loss of HLA-I gene expression was associated with repressive chromatin states including DNA methylation, histone H3 tri-methylation at lysine 27, and reduced chromatin accessibility. Pharmacological DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibition decreased DNA methylation and increased H3 lysine 27 acetylation and resulted in re-expression of HLA-I on the surface of tumor cells. Re-expression of HLA-I on LNCaP cells by DNMT and HDAC inhibition increased activation of co-cultured prostate specific membrane antigen (PSMA)27-38-specific CD8+ T-cells. HLA-I expression is epigenetically regulated by functionally reversible DNA methylation and chromatin modifications in human prostate cancer. Methylated HLA-I was detected in HLA-Ilow circulating tumor cells (CTCs), which may serve as a minimally invasive biomarker for identifying patients who would benefit from epigenetic targeted therapies.