Influence of glycan structure on the colonization of Streptococcus pneumoniae on human respiratory epithelial cells.

Virtually all living cells are encased in glycans. They perform key cellular functions such as immunomodulation and cell-cell recognition. Yet, how their composition and configuration affect their functions remains enigmatic. Here, we constructed isogenic capsule-switch mutants harboring 84 types of capsular polysaccharides (CPSs) in Streptococcus pneumoniae. This collection enables us to systematically measure the affinity of structurally related CPSs to primary human nasal and bronchial epithelial cells. Contrary to the paradigm, the surface charge does not appreciably affect epithelial cell binding. Factors that affect adhesion to respiratory cells include the number of rhamnose residues and the presence of human-like glycomotifs in CPS. Besides, pneumococcal colonization stimulated the production of interleukin 6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), and monocyte chemoattractantprotein-1 (MCP-1) in nasal epithelial cells, which also appears to be dependent on the serotype. Together, our results reveal glycomotifs of surface polysaccharides that are likely to be important for colonization and survival in the human airway.

Growth hormone-releasing hormone receptor antagonist MIA-602 attenuates cardiopulmonary injury induced by BSL-2 rVSV-SARS-CoV-2 in hACE2 mice.

COVID-19 pneumonia causes acute lung injury and acute respiratory distress syndrome (ALI/ARDS) characterized by early pulmonary endothelial and epithelial injuries with altered pulmonary diffusing capacity and obstructive or restrictive physiology. Growth hormone-releasing hormone receptor (GHRH-R) is expressed in the lung and heart. GHRH-R antagonist, MIA-602, has been reported to modulate immune responses to bleomycin lung injury and inflammation in granulomatous sarcoidosis. We hypothesized that MIA-602 would attenuate rVSV-SARS-CoV-2-induced pulmonary dysfunction and heart injury in a BSL-2 mouse model. Male and female K18-hACE2tg mice were inoculated with SARS-CoV-2/USA-WA1/2020, BSL-2-compliant recombinant VSV-eGFP-SARS-CoV-2-Spike (rVSV-SARS-CoV-2), or PBS, and lung viral load, weight loss, histopathology, and gene expression were compared. K18-hACE2tg mice infected with rVSV-SARS-CoV-2 were treated daily with subcutaneous MIA-602 or vehicle and conscious, unrestrained plethysmography performed on days 0, 3, and 5 (n = 7 to 8). Five days after infection mice were killed, and blood and tissues collected for histopathology and protein/gene expression. Both native SARS-CoV-2 and rVSV-SARS-CoV-2 presented similar patterns of weight loss, infectivity (~60%), and histopathologic changes. Daily treatment with MIA-602 conferred weight recovery, reduced lung perivascular inflammation/pneumonia, and decreased lung/heart ICAM-1 expression compared to vehicle. MIA-602 rescued altered respiratory rate, increased expiratory parameters (Te, PEF, EEP), and normalized airflow parameters (Penh and Rpef) compared to vehicle, consistent with decreased airway inflammation. RNASeq followed by protein analysis revealed heightened levels of inflammation and end-stage necroptosis markers, including ZBP1 and pMLKL induced by rVSV-SARS-CoV-2, that were normalized by MIA-602 treatment, consistent with an anti-inflammatory and pro-survival mechanism of action in this preclinical model of COVID-19 pneumonia.

Is ChatGPT 'ready' to be a learning tool for medical undergraduates and will it perform equally in different subjects? Comparative study of ChatGPT performance in tutorial and case-based learning questions in physiology and biochemistry.

PURPOSE: Generative AI will become an integral part of education in future. The potential of this technology in different disciplines should be identified to promote effective adoption. This study evaluated the performance of ChatGPT in tutorial and case-based learning questions in physiology and biochemistry for medical undergraduates. Our study mainly focused on the performance of GPT-3.5 version while a subgroup was comparatively assessed on GPT-3.5 and GPT-4 performances. MATERIALS AND METHODS: Answers were generated in GPT-3.5 for 44 modified essay questions (MEQs) in physiology and 43 MEQs in biochemistry. Each answer was graded by two independent examiners. Subsequently, a subset of 15 questions from each subject were selected to represent different score categories of the GPT-3.5 answers; responses were generated in GPT-4, and graded. RESULTS: The mean score for physiology answers was 74.7 (SD 25.96). GPT-3.5 demonstrated a statistically significant (p = .009) superior performance in lower-order questions of Bloom's taxonomy in comparison to higher-order questions. Deficiencies in the application of physiological principles in clinical context were noted as a drawback. Scores in biochemistry were relatively lower with a mean score of 59.3 (SD 26.9) for GPT-3.5. There was no statistically significant difference in the scores for higher and lower-order questions of Bloom's taxonomy. The deficiencies highlighted were lack of in-depth explanations and precision. The subset of questions where the GPT-4 and GPT-3.5 were compared demonstrated a better overall performance in GPT-4 responses in both subjects. This difference between the GPT-3.5 and GPT-4 performance was statistically significant in biochemistry but not in physiology. CONCLUSIONS: The differences in performance across the two versions, GPT-3.5 and GPT-4 across the disciplines are noteworthy. Educators and students should understand the strengths and limitations of this technology in different fields to effectively integrate this technology into teaching and learning.

Efficacy of adjunctive octenidine hydrochloride as compared to chlorhexidine and placebo as adjuncts to instrumentation in stage I-II periodontitis: A double-blinded randomized controlled trial.

OBJECTIVES: This randomized, controlled, double-blinded clinical trial aimed to evaluate the efficacy of octenidine hydrochloride and chlorhexidine mouthwashes as adjuncts to instrumentation in stage I-II periodontitis patients. METHODS: Forty-five patients with mild-to-moderate periodontitis were randomly allocated to three groups: 0.1% octenidine dihydrochloride (OCT), placebo, and 0.12% chlorhexidine (CHX) mouthwashes. Patients were instructed to use the mouthwash after instrumentation for twice a day up to 3 weeks. Periodontal parameters such as probing pocket depth (PPD), clinical attachment loss (CAL), O'Leary plaque index (PI), Loe and Silness gingival index (GI), Lobene stain index (SI), and oral soft tissue changes were recorded at baseline and once every week for 3 weeks. The visual analogue scale (VAS) was also recorded as a self-administered questionnaire at the end of the study. The one-way ANOVA was used to compare VAS scores between the groups. The repeated measures ANOVA and post hoc Newman-Keuls tests were used to assess the differences in the periodontal parameters between groups at different time intervals. The Kruskal-Wallis test was used to compare the mean SI. RESULTS: There was a significant reduction in the mean GI of the OCT and CHX groups compared to placebo (p < 0.05). OCT usage resulted in significantly less staining, according to mean SI, when compared to CHX. Furthermore, VAS scores revealed that OCT was significantly the preferred mouthwash (p < 0.01). CONCLUSION: Adjunctive octenidine hydrochloride may be an alternative to chlorhexidine in its ability to control the periodontal parameters in patients with stage I-II periodontitis. Further larger studies are necessary to confirm these findings.

Mobile application-assisted graded exercise practical: a remote teaching strategy to promote motivation and experiential learning in exercise physiology.

Practical classes are critical instructional activities in facilitating learning and motivation in health sciences education. With increasing pedagogical activities being conducted in virtual or remote settings, this study assessed how a remote practical assisted by physiological monitoring smartphone applications impacted student motivation and the achievement of intended learning outcomes in exercise physiology teaching. A total of 24 students (out of 30; 80%) were surveyed via a mixed-methods questionnaire containing 27 closed-ended, and 3 the traditional in-class practical in randomized order. Unpaired Student's t tests were performed for comparisons between interventions with a significance level set at P < 0.05. Students reported that both remote and in-class practicals strongly facilitated the achievement of learning outcomes. Self-reported scores for student satisfaction and perceived achievement of learning outcomes were similar between the two practical methodologies. Student motivation scores assessed using the Lab Motivation Scale revealed that students were more motivated during the remote practical, particularly in the effort domain (P < 0.05). This was in line with the identified themes from the qualitative responses that indicated that the remote practical was more engaging than the in-person practical, with greater opportunities for experiential learning and class involvement being the main factors underlying these findings. Taken together, remote practicals can be critical aspects of a blended learning curriculum that encourages student engagement and experiential learning. With further advancements in physiological monitoring wearables and smartphone technologies, remote practicals can be potential alternatives to traditional in-person practicals in exercise physiology teaching.NEW & NOTEWORTHY Remote practical classes, supported by physiological monitoring smartphone applications, were assessed for their utility in facilitating learning and raising student motivation in health sciences education in this study. A comparison of remote practicals with traditional in-class practicals revealed that a remote practical is an effective method for reinforcing physiology learning objectives with the added advantage of increased student motivation. The added value of remote practicals may be attributed to more experiential learning opportunities and increased engagement levels.

Very-Early Onset Chronic Active Colitis with Heterozygous Variants in LRBA1 and CARD11, a Case of "Immune TOR-Opathies".

BACKGROUND: A small subset of cases of inflammatory bowel disease (IBD) occurs as a result of single gene defects, and typically occurs in young or very young pediatric patients, referred to as "monogenic very-early onset IBD (VEO-IBD)". The gene variants leading to monogenic VEO-IBD are often associated with primary immunodeficiency syndromes. CASE REPORT: A six year-old girl presented to our gastroenterology clinic with LRBA deficiency with a heterozygous mutation at c.1399 A > G, p Met467Val, histopathologic chronic active colitis without granulomas and clinical chronic colitis. Her gastrointestinal symptoms began at age 5 with bloody diarrhea, abdominal pain and weight loss. Whole exome sequencing revealed a CARD11 heterozygous de novo mutation (c.220 + 1G > A). She was in clinical remission on only abatacept. DISCUSSION: We present a case of monogenic VEO-IBD associated with two heterozygous variants in LRBA1 and CARD11, both considered as key players in the newly proposed "immune TOR-opathies".

Deep Phenotyping by Mass Cytometry and Single-Cell RNA-Sequencing Reveals LYN-Regulated Signaling Profiles Underlying Monocyte Subset Heterogeneity and Lifespan.

RATIONALE: Monocytes are key effectors of the mononuclear phagocyte system, playing critical roles in regulating tissue homeostasis and coordinating inflammatory reactions, including those involved in chronic inflammatory diseases such as atherosclerosis. Monocytes have traditionally been divided into 2 major subsets termed conventional monocytes and patrolling monocytes (pMo) but recent systems immunology approaches have identified marked heterogeneity within these cells, and much of what regulates monocyte population homeostasis remains unknown. We and others have previously identified LYN tyrosine kinase as a key negative regulator of myeloid cell biology; however, LYN's role in regulating specific monocyte subset homeostasis has not been investigated. OBJECTIVE: We sought to comprehensively profile monocytes to elucidate the underlying heterogeneity within monocytes and dissect how Lyn deficiency affects monocyte subset composition, signaling, and gene expression. We further tested the biological significance of these findings in a model of atherosclerosis. METHODS AND RESULTS: Mass cytometric analysis of monocyte subsets and signaling pathway activation patterns in conventional monocytes and pMos revealed distinct baseline signaling profiles and far greater heterogeneity than previously described. Lyn deficiency led to a selective expansion of pMos and alterations in specific signaling pathways within these cells, revealing a critical role for LYN in pMo physiology. LYN's role in regulating pMos was cell-intrinsic and correlated with an increased circulating half-life of Lyn-deficient pMos. Furthermore, single-cell RNA sequencing revealed marked perturbations in the gene expression profiles of Lyn-/- monocytes with upregulation of genes involved in pMo development, survival, and function. Lyn deficiency also led to a significant increase in aorta-associated pMos and protected Ldlr-/- mice from high-fat diet-induced atherosclerosis. CONCLUSIONS: Together our data identify LYN as a key regulator of pMo development and a potential therapeutic target in inflammatory diseases regulated by pMos.

Incidence of Head and Neck Cancers before and after 9/11 in New York City and New York State.

INTRODUCTION: The incidence of cancers in New York State (NYS) before and after 9/11 including lung, colorectal, and renal cancers has been previously described. To date, the incidence of head and neck cancers (HNCs) before and after 9/11 has not been described. METHODS: Cancers involving the oral cavity and oropharynx; the nose, nasal cavity, nasopharynx, and middle ear; larynx; and thyroid were identified using the New York State Cancer Registry (NYSCR). Age-adjusted incidence and rates per 100,000 residents from 1987 to 2015 were analyzed using joinpoint regression. Trends in incidence using annual percent changes are presented. RESULTS: The overall rate of HNC increased slightly by 0.7% (p < 0.001) from 1987 to 2003 in NYS. From 2003 to 2008, the rate increased by 5.73% (p < 0.001), and from 2008 to 2015, the rate increased by 1.68% (p < 0.001). The rate of thyroid cancer increased by 6.79% (p < 0.001) from 1987 to 2003, by 9.99% (p < 0.001) from 2003 to 2009, and by 2.41% (p = 0.001) from 2009 to 2015. The rate of thyroid cancer was higher in women at all time points. In a subset analysis of HNCs excluding thyroid cancer, the rate decreased by 2.02% (p < 0.001) from 1991 to 2001, followed by a nonsignificant increase of 0.1% (p = 0.515) from 2001 to 2015. The rate of oropharyngeal (OP) cancer significantly increased from 1999 to 2015 (2.65%; p < 0.001). The rate of oral cavity cancer significantly decreased from 1987 to 2003 (1.97%; p < 0.001), with no significant change after 2003. The rate of laryngeal cancer decreased significantly by 2.43% (p < 0.001) from 1987 to 2015, as did the rate of nasal cavity/nasopharyngeal cancer (0.33%; p = 0.03). CONCLUSIONS: In NYS, OP cancer and thyroid cancer rates increased significantly during the study period. The rate of thyroid cancer was higher in women. The rate of combined HNC increased significantly after 9/11 compared to before 9/11; however, in a subset analysis of all HNC patients excluding thyroid cancer, the rate decreased significantly prior to 9/11 and then nonsignificantly increased afterward. This suggests that the increase in thyroid cancer accounts for the increase in combined HNC in NYS. The impact of 9/11 on rates of HNC requires further research.

Quick and facile preparation of histone proteins from the green microalga Chlamydomonas reinhardtii and other photosynthetic organisms.

The development of universal, broadly applicable methods for histone extraction from animal cells and tissues has unlocked the ability to compare these epigenetic-influencing proteins across tissue types, healthy and diseased states, and cancerous versus normal cells. However, for plants and green algae, a quick and easily implemented histone extraction method has yet to be developed. Here, we report an optimized method that provides a unified approach to extract histones for the green microalgal species Chlamydomonas reinhardtii and Scenedesmus dimorphus as well as for maize (corn) leaf tissue. Histone extraction methods include treatment with high salt concentrations and acidification. Preparations of nuclei can be made in ∼3.5 h and histones extracted in ∼3.5 h either immediately or nuclei may be frozen and histone proteins can be later extracted without a change in histone PTM patterns. To examine the efficiency of the new methods provided, we performed both qualitative and quantitative analysis of salt and acid-extracted whole histone proteins (SAEWH) via SDS-PAGE gel electrophoresis and intact protein mass spectrometry. SDS-PAGE analysis indicated that histone yields decrease when using walled Chlamydomonas strains relative to cell-wall-less mutants. Using top-down mass spectrometry (TDMS) for intact protein analysis, we confirmed the presence of H4K79me1 in multiple algal species; however, this unique modification was not identified in corn leaf tissue and has not been reported elsewhere. TDMS measurements of SAEWH extracts also revealed that oxidation which occurs during the histone extraction process does not increase with exposure of harvested algal cells, their nuclei and the extracted histone samples to light.

Tetraspanins: Host Factors in Viral Infections.

Tetraspanins are transmembrane glycoproteins that have been shown increasing interest as host factors in infectious diseases. In particular, they were implicated in the pathogenesis of both non-enveloped (human papillomavirus (HPV)) and enveloped (human immunodeficiency virus (HIV), Zika, influenza A virus, (IAV), and coronavirus) viruses through multiple stages of infection, from the initial cell membrane attachment to the syncytium formation and viral particle release. However, the mechanisms by which different tetraspanins mediate their effects vary. This review aimed to compare and contrast the role of tetraspanins in the life cycles of HPV, HIV, Zika, IAV, and coronavirus viruses, which cause the most significant health and economic burdens to society. In doing so, a better understanding of the relative contribution of tetraspanins in virus infection will allow for a more targeted approach in the treatment of these diseases.

Acquisition time, radiation dose, subjective and objective image quality of dual-source CT scanners in acute pulmonary embolism: a comparative study.

OBJECTIVES: To compare the scan acquisition time, radiation dose, subjective and objective image quality of two dual-source CT scanners (DSCT) for detection of acute pulmonary embolism. METHODS: Two hundred twenty-one scans performed on the 2nd-generation DSCT and 354 scans on the 3rd-generation DSCT were included in this large retrospective study. In a randomized blinded design, two radiologists independently reviewed the scans using a 5-point Likert scale. Radiation dose and objective image quality parameters were calculated. RESULTS: Mean acquisition time was significantly lower in the 3rd-generation DSCT (2.81 s ± 0.1 in comparison with 9.7 s ± 0.15 [mean ± SD] respectively; p < 0.0001) with the 3rd generation 3.4 times faster. The mean subjective image quality score was 4.33/5 and 4/5 for the 3rd- and 2nd-generation DSCT respectively (p < 0.0001) with strong interobserver reliability agreement. DLP, CTDIvol, and ED were significantly lower in the 3rd than the 2nd generation (175.6 ± 63.7 mGy cm; 5.3 ± 1.9 mGy and 2.8 ± 1.2 mSv in comparison with 266 ± 255 mGy.cm; 7.8 ± 2.2 mGy and 3.8 ± 4.3 mSv). Noise was significantly lower in the 3rd generation (p < 0.01). Signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and figure of merit (FOM), a dose-insensitive index for CNR, were significantly higher in the 3rd-generation DSCT (33.5 ± 23.4; 29.0 ± 21.3 and 543.7 ± 1037 in comparison with 23.4 ± 17.7; 19.4 ± 16.0 and 170.5 ± 284.3). CONCLUSION: Objective and subjective image quality are significantly higher on the 3rd-generation DSCT with significantly lower mean acquisition time and radiation dose. KEY POINTS: • The 3rd-generation DSCT scanner provides an improved image quality, less perceived artifacts, and lower radiation dose in comparison with the 2nd-generation DSCT, when operating in dual-energy (DE) mode. • The 3.4-times-faster 3rd-generation DSCT scanner can be of particular value in patients with chronic lung diseases or breathing difficulties that prevent adequate breathhold.

The paralyzing legal costs of facial nerve injury in head and neck tumors.

PURPOSE: Facial nerve paralysis from head and neck tumors can result from disease progression or iatrogenic causes, leading to litigation. The aim of this study was to investigate lawsuits regarding facial paralysis as a consequence of these tumors to understand and better educate physicians behind the reasons for litigation. METHODS: Jury verdict reviews were obtained from the Westlaw database from 1985 to 2018. Gathered data, including verdicts, litigation reasons, defendant specialties, and amounts awarded, were analyzed via Statistical Package for the Social Sciences. RESULTS: Of the 26 lawsuits analyzed, the leading reason for litigation was failure to diagnose (53.8%), followed by iatrogenic injury (34.6%). The average award was $2,704,470. Otolaryngologists were the most common defendants. Defendants that included an otolaryngologist had shorter delays of diagnosis compared to those that did not (p < 0.05). CONCLUSION: Failure to diagnose parotid injury was the leading cause of litigation. In instances where the jury found for the plaintiff, the amount was material. There were equivalent incidences of cases in favor of plaintiffs and defendants.

Epithelial-Myoepithelial Carcinoma Presenting as a Pseudo Veno-Lymphatic Malformation.

A 41-year-old woman presented with several days of right eye pain and blurred vision. Examination and radiologic workup were consistent with a veno-lymphatic malformation, including demonstration of cystic fluid-fluid levels on imaging. Histopathology supported the presence of cystic spaces but revealed the final diagnosis of lacrimal epithelial-myoepithelial carcinoma, a rare neoplasm typically associated with the salivary gland. This represents the first reported case of lacrimal epithelial-myoepithelial carcinoma presenting with pain and evidence of intralesional cysts on radiographic and histopathologic evaluation. Better characterization will enable recognition, complete surgical excision, and potential adjuvant radiation therapy.

Reliability and validity of the DBT-VLCS: A measure to code validation strategies in dialectical behavior therapy sessions.

OBJECTIVE: There are six strategies or validation levels in dialectical behavior therapy (DBT), yet there are no measures designed to code for them. This absence limits our understanding of the relationship between validation strategies and treatment outcome. The DBT-Validation Level Coding Scale (DBT-VLCS) was developed to overcome this limitation. METHOD: This research reports on the interrater reliability and content validity for the DBT-VLCS. RESULTS: Overall, interrater reliability was excellent for all items, with the exception of two items that demonstrated good reliability. Good content validity was demonstrated for six of the seven items. CONCLUSIONS: This preliminary study suggests that the DBT-VLCS is a reliable and valid measure to code the presence of validation in DBT. This measure creates the opportunity for research that has not previously been possible.

Outcomes of Chandelier-Assisted Scleral Buckling in Rhegmatogenous Retinal Detachments: Systematic Review and Meta-analysis.

Purpose: To examine the outcomes of chandelier endoillumination-assisted scleral buckling (chandelier scleral buckling) for rhegmatogenous retinal detachments (RRDs) and compare them with those of standard scleral buckling using indirect ophthalmoscopy. Methods: A literature search was performed on April 15, 2023. Outcomes analyzed included the primary anatomic success rates, surgical duration, and complication rates. A meta-analysis of proportions estimated the pooled success rate of chandelier scleral buckling. In addition, meta-analyses compared the success rates between pseudophakic eyes and phakic eyes having chandelier scleral buckling and compared success rates and surgical duration between standard scleral buckling and chandelier scleral buckling. Results: Thirty studies with 1133 eyes were included. The pooled primary anatomic success rate of chandelier scleral buckling was 91.7% (95% CI, 89.6%-93.6%). In studies comparing success rates between the 2 techniques, there was no significant difference (risk ratio, 1.01; 95% CI, 0.94-1.08; P = .80). The surgical times were significantly shorter with chandelier scleral buckling than with standard scleral buckling (mean difference, -18.83; 95% CI, -30.88 to -6.79; P = .002). There was no significant difference in the success rate between pseudophakic eyes and phakic eyes (risk ratio, 0.99; 95% CI, 0.91-1.08; P = .89). No cases of endophthalmitis were reported. Conclusions: Chandelier endoillumination-assisted scleral buckling may be a promising technique given its high rate of primary anatomic success for RRDs and success rates similar to those of standard scleral buckling. There was no significant difference in the efficacy of chandelier scleral buckling between pseudophakic eyes and phakic eyes.

180° versus 360° Selective Laser Trabeculoplasty in Open Angle Glaucoma and Ocular Hypertension: A Systematic Review and Meta-Analysis.

PRECIS: This systematic review and meta-analysis found that 360° selective laser trabeculoplasty (SLT) is significantly more effective than 180° SLT at reducing intraocular pressure at one month and one year follow-ups, without increased serious adverse event risk. PURPOSE: To determine the efficacy of 180° versus 360° selective laser trabeculoplasty (SLT) in adults with open angle glaucoma (OAG) and ocular hypertension (OHT). METHODS: A systematic review was performed using PubMed, Embase, and Scopus databases, from 1995 to December 30, 2023, for studies comparing 180° and 360° SLT in adults with OAG and OHT (PROSPERO ID: CRD42024497832). Meta-analyses were performed to calculate nominal percent and raw reductions in IOP between treatment groups at one-month, one-year, and two-year follow-ups, as well as success rates, defined as a 20% or greater IOP reduction. RESULTS: Nine studies with 1044 eyes were included; 491 received 180°SLT and 553 received 360° SLT. At the one-month follow-up, 360° SLT reduced IOP by 3.45% more (WMD=3.45; 95% CI: 2.02-4.88; P<0.00001) and 0.87 mmHg more (WMD=0.87; 95% CI: 0.35-1.38; P=0.0010). At the one-year follow-up, 360° SLT reduced IOP by 4.33% more (WMD=4.33; 95% CI: 2.35-6.32; P<0.0001) and 1.15 mmHg more (WMD=1.15; 95% CI: 0.25-2.04; P=0.01). At two years of follow up, 360° SLT reduced IOP by 4.86% more (WMD=4.86; 95% CI: -0.32, 10.0; P=0.07) and 1.25 mmHg more (WMD=1.25; 95% CI: -0.29, 2.79; P=0.11), however the difference was not statistically significant. Compared to 360° SLT, 180° SLT had a significantly lower success rate (OR=0.50; 95% CI: 0.35-0.72; P=0.0002). There was no difference in serious complications between interventions. CONCLUSION: 360° SLT is more effective than 180° SLT at lowering IOP at one-month and one-year follow-ups as well as achieving successful IOP control without increased risk of serious complications.

Transmucosal administration of pentobarbital and phenytoin solution induces euthanasia in bearded dragons (Pogona vitticeps).

OBJECTIVE: To assess the efficacy of transmucosal euthanasia solution to induce euthanasia. ANIMALS: 6 bearded dragons (Pogona vitticeps). METHODS: An initial dose of euthanasia solution containing pentobarbital and phenytoin sodium was administered transmucosally in conscious lizards (100 mg/kg pentobarbital dose), followed by a second dose 20 minutes later (400 mg/kg pentobarbital dose). The presence of movement, leakage of euthanasia solution, behaviors consistent with oral irritation, respiratory rate, heart rate, palpebral and corneal reflex, and response to noxious stimuli were recorded until death, confirmed by the absence of Doppler cardiac flow and cardiac electrical activity. The time to loss of all parameters was calculated. Postmortem evaluation allowed for histopathologic evaluation of the oral cavity and gastrointestinal tract to detect potential mucosal damage from the alkaline euthanasia solution. RESULTS: The median time to death was 300 minutes (range, 300 to 360 minutes), median time to respiratory arrest was 30 minutes (range, 30 to 50 minutes), and median time to loss of deep pain response was 30 minutes (range, 20 to 50 minutes). Signs consistent with oral irritation occurred in 4 of 6 (66.7%) lizards, including 2 lizards that exhibited whole-body spasms after euthanasia solution administration. Histopathologic changes indicating peracute mucosal ulceration, suspected to be from caustic causes, were identified in 1 (1/6 [16.7%]) lizard. CLINICAL RELEVANCE: Transmucosal euthanasia solution administration resulted in clinical euthanasia within 6 hours. This method should be utilized only after premedication with analgesic and/or anesthetic medications due to the potential for acute mucosal ulceration and behaviors that may be distressing in client-owned animals.

Multiregion sampling of de novo metastatic prostate cancer reveals complex polyclonality and augments clinical genotyping.

De novo metastatic prostate cancer is highly aggressive, but the paucity of routinely collected tissue has hindered genomic stratification and precision oncology. Here, we leveraged a rare study of surgical intervention in 43 de novo metastatic prostate cancers to assess somatic genotypes across 607 synchronous primary and metastatic tissue regions plus circulating tumor DNA. Intra-prostate heterogeneity was pervasive and impacted clinically relevant genes, resulting in discordant genotypes between select primary restricted regions and synchronous metastases. Additional complexity was driven by polyclonal metastatic seeding from phylogenetically related primary populations. When simulating clinical practice relying on a single tissue region, genomic heterogeneity plus variable tumor fraction across samples caused inaccurate genotyping of dominant disease; however, pooling extracted DNA from multiple biopsy cores before sequencing can rescue misassigned somatic genotypes. Our results define the relationship between synchronous treatment-sensitive primary and metastatic lesions in men with de novo metastatic prostate cancer and provide a framework for implementing genomics-guided patient management.

Emotional cascades and self-injury: investigating instability of rumination and negative emotion.

OBJECTIVE: Nonsuicidal self-injury (NSSI) is a public health concern and risk factor for suicide. The Emotional Cascade Model (ECM) proposes that NSSI partially functions as a distraction from cascades of negative affect and rumination. The purpose of this study was to examine the roles of trait rumination, and momentary instability of rumination and negative emotion, in NSSI. METHOD: Experience sampling methods were used to monitor thoughts, emotions, and behaviors in 47 individuals reporting dysregulated behaviors including NSSI. Instability indices were generated for rumination and negative emotion using the momentary assessments. RESULTS: Twenty-five episodes of NSSI were reported during monitoring. Trait rumination prospectively predicted NSSI episodes, and the instability indices interacted to predict NSSI. CONCLUSIONS: Consistent with the ECM, the interaction between rumination instability and negative affect instability during monitoring significantly predicted NSSI, with the strongest effects occurring for sadness and rumination about past. These findings may enhance conceptualization and treatment of patients with NSSI.

Detection and resection of carcinoma in situ of the bladder: Implications for clinical trial design.

INTRODUCTION: The path to approval of novel therapeutics for patients with BCG-unresponsive non-muscle invasive bladder cancer (NMIBC) requires demonstration of efficacy in eradicating carcinoma in situ (CIS), as determined by cytology, white light cystoscopy and only sometimes mandatory re-biopsy. This paradigm is based on the premise that CIS, in contrast to papillary tumors, cannot be completely resected. We aimed to determine the accuracy of CIS by standard means and the rate at which CIS may be eradicated by transurethral bladder tumor resection (TURBT). METHODS: We performed a retrospective analysis of consecutive patients who underwent radical cystectomy (RC) for high risk NMIBC or muscle invasive bladder cancer (MIBC) between 2005 and 2019 in a tertiary academic center. The concordance in the presence of CIS in matched TURBT and RC samples was calculated. RESULTS: Complete pathologic information was available for 816 patients with urothelial carcinoma. CIS was detected at TURBT in 354 (43.4%) patients (64.0% NMIBC, 32.3% MIBC) and at RC in 436 (53.4%) patients (64.7% NMIBC, 47.4% MIBC). CIS was missed by TURBT in 199 (45.6%) of those cases (NMIBC 25.4%, MIBC 60.6%). CIS detected on TURBT was not found in the RC specimen in 33.1% (117/354) of cases. Lack of prospective bladder mapping and central pathology review are limitations. CONCLUSION: Our results suggest that TURBT is inaccurate in detecting CIS. The absence of CIS in the RC specimen after detection in the matched TURBT specimen suggests that CIS may be completely resected by TURBT in a proportion of patients. These factors need to be considered in the design of clinical trials in patients with NMIBC. The use of random biopsies or enhanced cystoscopy could improve the accuracy of CIS detection, but the former is associated with patient morbidity and randomization would alleviate concern about these variables impacting clinical trial outcomes.