Atypical B cells promote cancer progression and poor response to Bacillus Calmette-Guérin in non-muscle invasive bladder cancer.

Poor response to Bacillus Calmette-Guérin (BCG) immunotherapy remains a major barrier in the management of patients with non-muscle invasive bladder cancer (NMIBC). Multiple factors are associated with poor outcomes, including biological aging and female sex. More recently, it has emerged that a B-cell infiltrated pre-treatment immune microenvironment of NMIBC tumors can influence the response to intra-vesically administered BCG. The mechanisms underlying the roles of B cells in NMIBC are poorly understood. Here, we show that B-cell dominant tertiary lymphoid structures (TLSs), a hallmark feature of the chronic mucosal immune response, are abundant and located close to the epithelial compartment in pre-treatment tumors from BCG non-responders. Digital spatial proteomic profiling of whole tumor sections from male and female patients with NMIBC who underwent treatment with intravesical BCG, revealed higher expression of immune exhaustion-associated proteins within the tumor-adjacent TLSs in both responders and non-responders. Chronic local inflammation, induced by the N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen, led to TLS formation with recruitment and differentiation of the immunosuppressive atypical B-cell (ABC) subset within the bladder microenvironment, predominantly in aging female mice compared to their male counterparts. Depletion of ABCs simultaneous to BCG treatment delayed cancer progression in female mice. Our findings provide evidence indicating a role for ABCs in BCG response and will inform future development of therapies targeting the B cell-exhaustion axis.

Virtual OSCE examinations during COVID-19 A 360 satisfaction assessment from examiners and candidates.

INTRODUCTION: We sought to determine the satisfaction rates of examiners and candidates in a virtual Objective Structured Clinical Exam (OSCE) of graduating Canadian urology residents. METHODS: An annual mock exam, aimed at simulating the licencing urology exam for Canadian graduates, was moved to an online format for the 2020 cohort. This exam consists of an OSCE, and a written multiple-choice exam. The Telemedicine Satisfaction Questionnaire (TSQ), a previously validated tool for clinical encounters with three sub-domains (quality of care provided, similarity to face-to-face encounter, and perception of the interaction) was modified for the purposes of evaluating the OSCE encounter. The TSQ was sent electronically to all examiners and candidates after the exam. RESULTS: There were 14/16 responses from examiners (87.5%) and 24/39 responses from candidates (61.5%). Overall, the format was judged to be a good experience by 13/14 (92.9%) of examiners and 21/24 (87.5%) of candidates; however, when asked specifically if the virtual OSCE was an acceptable way to determine a candidate's competency to practice urology independently, only 8/14 (57.1%) of examiners and 15/24 (62.5%) of candidates agreed. CONCLUSIONS: This study demonstrates an overall good satisfaction rate among both examiners and candidates when using a teleconference format for a mock OSCE.

Renal colic imaging practice patterns in Ontario A population-based study.

INTRODUCTION: Computed tomography (CT) is associated with increased cost and exposure to radiation when compared to ultrasound (US) in patients with renal colic. Consequently, a 2014 Choosing Wisely recommendation states US should be used over CT in uncomplicated presentations in patients under age 50. The objective of this study was to describe imaging practice patterns in Ontario among patients presenting with renal colic and the relationship between initial imaging modality, subsequent imaging, and burden of care indicators. METHODS: This is a population-based study of patients who presented with renal colic in Ontario from 2003-2019 using administrative data. Patients were assessed according to their first imaging modality during their index visit. Descriptive statistics and Chi-squared test were used to examine differences between these groups. The primary outcome was the need for subsequent imaging. Secondary outcomes were length of renal colic episode, days to surgery, and number of emergency department (ED ) and primary care visits during the renal colic episode. Univariate and multivariable logistic regression models were used. RESULTS: A total of 429 060 patients were included in the final analysis. Of those, 50.5% (216 747) had CT as their initial imaging modality, 20% (84 672) had US, and 3% (13 643) had both on the same day. Subsequent imaging was obtained in 40.7% of those who had CT as the initial imaging, compared to 43% in those who had US and 43% who had both. Of those who initially had an US, 38% went on to have at least one CT during their renal colic episode, including those who had CT on the same day as initial US, while 62% were able to avoid CT altogether. In contrast, 17% had a repeat CT after an initial CT at the time of presentation. The overall use of US increased from 15% to 31% during the study period. The length of the renal colic episode was slightly longer in those who had a CT first compared to US in multivariable models (adjusted risk ratio [ARR ] 1.005, 95% confidence interval [CI] 1.000-1.009); however, the time to surgery was less in those who had a CT first (ARR 0.831, 95% CI 0.807-0.856). Fewer ED and family physician visits were seen in those who had an initial CT. CONCLUSIONS: In patients with renal colic in Ontario, approximately half have CT as the initial imaging modality despite US being recommended in uncomplicated presentations. While US use remains low, its use doubled during this study period, demonstrating an encouraging trend. Those who have US first can often avoid subsequent CT.

Impact of chronic variable stress on neuroendocrine hypothalamus and pituitary in male and female C57BL/6J mice.

Chronic stress exerts multiple negative effects on the physiology and health of an individual. In the present study, we examined hypothalamic, pituitary and endocrine responses to 14 days of chronic variable stress (CVS) in male and female C57BL/6J mice. In both sexes, CVS induced a significant decrease in body weight and enhanced the acute corticosterone stress response, which was accompanied by a reduction in thymus weight only in females. However, single-point blood measurements of basal prolactin, thyroid-stimulating hormone, luteinising hormone, growth hormone and corticosterone levels taken at the end of the CVS were not different from those of controls. Similarly, pituitary mRNA expression of Fshb, Lhb, Prl and Gh was unchanged by CVS, although Pomc and Tsh were significantly elevated. Within the adrenal medulla, mRNA for Th, Vip and Gal were elevated following CVS. Avp transcript levels within the paraventricular nucleus of the hypothalamus were increased by CVS; however, levels of Gnrh1, Crh, Oxt, Sst, Trh, Ghrh, Th and Kiss1 remained unchanged. Oestrous cycles were lengthened slightly by CVS and ovarian histology revealed a reduction in the number of preovulatory follicles and corpora lutea. Taken together, these observations indicate that 14 days of CVS induces an up-regulation of the neuroendocrine stress axis and creates a mild disruption of female reproductive function. However, the lack of changes in other neuroendocrine axes controlling anterior and posterior pituitary secretion suggest that most neuroendocrine axes are relatively resilient to CVS.

MCM complex members MCM3 and MCM7 are associated with a phenotypic spectrum from Meier-Gorlin syndrome to lipodystrophy and adrenal insufficiency.

The MCM2-7 helicase is a heterohexameric complex with essential roles as part of both the pre-replication and pre-initiation complexes in the early stages of DNA replication. Meier-Gorlin syndrome, a rare primordial dwarfism, is strongly associated with disruption to the pre-replication complex, including a single case described with variants in MCM5. Conversely, a biallelic pathogenic variant in MCM4 underlies immune deficiency with growth retardation, features also seen in individuals with pathogenic variants in other pre-initiation complex encoding genes such as GINS1, MCM10, and POLE. Through exome and chromium genome sequencing, supported by functional studies, we identify biallelic pathogenic variants in MCM7 and a strong candidate biallelic pathogenic variant in MCM3. We confirm variants in MCM7 are deleterious and through interfering with MCM complex formation, impact efficiency of S phase progression. The associated phenotypes are striking; one patient has typical Meier-Gorlin syndrome, whereas the second case has a multi-system disorder with neonatal progeroid appearance, lipodystrophy and adrenal insufficiency. We provide further insight into the developmental complexity of disrupted MCM function, highlighted by two patients with a similar variant profile in MCM7 but disparate clinical features. Our results build on other genetic findings linked to disruption of the pre-replication and pre-initiation complexes, and the replisome, and expand the complex clinical genetics landscape emerging due to disruption of DNA replication.

Cavernous hemangioma of the adrenal gland mimicking a hepatic hemangioma.

Adrenal cavernous hemangiomas are a rare, benign, and non-functional tumor. We report a case of a 62-year-old male who presented with right upper quadrant and flank pain. Physical examination revealed a fullness of the right upper quadrant. Both computed tomography and magnetic resonance imaging suggested a hemangioma originating from the liver. During angiography with the intent of embolization, it was discovered that the vascular supply was consistent with an adrenal mass rather than a hepatic origin. The patient was referred to Urology and underwent curative right open adrenalectomy and nephrectomy. Histopathology confirmed the diagnosis of an adrenal cavernous hemangioma.

Biallelic variants in SLC35C1 as a cause of isolated short stature with intellectual disability.

Variants in SLC35C1 underlie leucocyte adhesion deficiency (LADII) or congenital disorder of glycosylation type 2c (CDGIIc), an autosomal recessive disorder of fucosylation. This immunodeficiency syndrome is generally characterized by severe recurrent infections, Bombay blood group, reduced growth and intellectual disability (ID). Features are all caused by an inability to generate key fucosylated molecules due to a defective transport of GDP-fucose into the Golgi. Here we report the use of exome sequencing to identify biallelic variants in SLC35C1 (c.501_503delCTT, p.(Phe168del) and c.891T > G, p.(Asn297Lys)) in an individual with short stature and ID. Retrospective clinical examination based on the genetic findings revealed increased otitis media as the only immunological feature present in this child. Biochemical analysis of patient serum identified a clear but mild decrease in protein fucosylation. Modelling all described missense mutations on a SLC35C1 protein model showed pathogenic substitutions localise to close to the dimer interface, providing insight into the possible pathophysiology of non-synonymous causative variants identified in patients. Our evidence confirms this is the second family presenting with only a subset of features and broadens the clinical presentation of this syndrome. Of note, both families segregated a common allele (p.Phe168del), suggesting there could be an associated genotype-phenotype relationship for specific variants. Based on two out of 14 reported families not presenting with the characteristic features of SLC35C1-CDG, we suggest there is clinical utility in considering this gene in patients with short stature and ID.

Targeting Cognition and Motivation in Coordinated Specialty Care for Early Psychosis: A Grant Report.

In this grant report, we describe our project to expand measurement-based psychiatric care across 6 early psychosis treatment teams in Minnesota, and to provide a neuroscience-informed cognitive training and motivation enhancement program for individuals with early psychosis. This project is part of the NIMH Early Psychosis Intervention Network (EPINET) initiative which seeks to link data from treatment centers nationally that offer evidence-based specialty care to persons experiencing early psychosis. Systematic analyses of pooled data collected in EPINET will help inform methods for early psychosis care, psychosis risk factors, and pre-emptive interventions. As part of the national EPINET, our hub (Early Psychosis Intervention-Minnesota, EPI-MINN), will: (1) provide measurement-based care in coordinated specialty care programs for early psychosis, (2) determine whether a structured feedback report provides benefit to stakeholders-service users, family members, and primary clinicians, and (3) explore whether deficits in cognition and motivated behavior-two domains that significantly impact functioning and overall quality of life in early psychosis-can be addressed as key treatment goals by implementing a 12-week mobile intervention. Using a regression discontinuity design, participants will be randomized to the cognitive training and motivational enhancement intervention or to treatment as usual. The intervention consists of neuroscience-informed, computerized auditory and social cognitive training exercises, as well as a mobile app where participants interact with each other and with a motivational coach. Participants will complete assessments at 4 time points: baseline and post-intervention (i.e., at 6 months), and again at 12 and 18 months to test the long-term effects of the intervention. All assessments and interventions in this project can be completed entirely remotely.

Expanding the phenotypic spectrum associated with DPF2: A new case report.

Coffin-Siris syndrome (CSS) is a clinically and genetically heterogeneous developmental disorder, linked to disruption of the BAF chromatin-remodeling complex. Recently, de novo missense and truncating variants have been reported in DPF2 in patients sharing some of the common features of CSS. Here we report a further individual harboring a novel de novo missense DPF2 variant, c.1066T>G, p.Cys356Gly. Structural modeling indicated that the predicted amino acid substitution affects a core residue required for zinc ion coordination and would likely alter the PHD2 domain structure of DPF2. The clinical presentation of Pierre Robin sequence and diaphragmatic hernia did not immediately suggest CSS, with the more common CSS features of hypoplastic toenails and characteristic facial features very subtle. This individual further broadens the phenotypic features of DPF2-related CSS, as well as CSS more generally.

The Shame-Blame Game: Is It Still Necessary? A National Survey of Shame-based Teaching Practice in Canadian Plastic Surgery Programs.

BACKGROUND: As understanding of poor physician mental health and burnout strengthens, it is becoming important to identify factors that detract from wellbeing. Shame-based learning (SBL) is detrimental to psychological health and can contribute to burnout, substance abuse and suicide. This study endeavoured to quantify the unknown prevalence and effects of SBL in Canadian plastic surgery programs. METHODS: An electronic survey was sent to all attending surgeons and trainee (residents and fellows) members of the Canadian Society of Plastic Surgeons. SBL was assessed using a validated questionnaire. Data was analyzed using descriptive statistics and thematic analysis. RESULTS: 98 responses (14.7%) comprising of 63 attending surgeons and 36 trainees were received. The majority of attending surgeons (78 percent) and trainees (67%) have been shamed. Fourteen percent of trainees and 9% of attending surgeons felt that SBL is necessary. The most common event provoking shaming for trainees was wrong answers (56%) and for attending surgeons was disagreement in clinical care (21%). For both groups, shamers were in positions of authority. The most common effect of SBL in trainees was a loss of self-confidence (53%), compared to no negative effect in attending surgeons (49 percent). Thirty-nine percent of trainees dealt with shaming events with support from fellow trainees (39 percent), while attending surgeons kept it to themselves (40 percent). CONCLUSION: SBL is present in Canadian plastic surgery residency programs and has numerous detrimental effects. To foster better mental health, residency programs should identify ongoing SBL and make efforts to transition to healthier feedback strategies.

Interleukin-6-mediated signaling in adrenal medullary chromaffin cells.

The pro-inflammatory cytokines, tumor necrosis factor-α, and interleukin-1ß/α modulate catecholamine secretion, and long-term gene regulation, in chromaffin cells of the adrenal medulla. Since interleukin-6 (IL6) also plays a key integrative role during inflammation, we have examined its ability to affect both tyrosine hydroxylase activity and adrenomedullary gene transcription in cultured bovine chromaffin cells. IL6 caused acute tyrosine/threonine phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2), and serine/tyrosine phosphorylation of signal transducer and activator of transcription 3 (STAT3). Consistent with ERK1/2 activation, IL6 rapidly increased tyrosine hydroxylase phosphorylation (serine-31) and activity, as well as up-regulated genes, encoding secreted proteins including galanin, vasoactive intestinal peptide, gastrin-releasing peptide, and parathyroid hormone-like hormone. The effects of IL6 on the entire bovine chromaffin cell transcriptome were compared to those generated by G-protein-coupled receptor (GPCR) agonists (histamine and pituitary adenylate cyclase-activating polypeptide) and the cytokine receptor agonists (interferon-α and tumor necrosis factor-α). Of 90 genes up-regulated by IL6, only 16 are known targets of IL6 in the immune system. Those remaining likely represent a combination of novel IL6/STAT3 targets, ERK1/2 targets and, potentially, IL6-dependent genes activated by IL6-induced transcription factors, such as hypoxia-inducible factor 1α. Notably, genes induced by IL6 include both neuroendocrine-specific genes activated by GPCR agonists, and transcripts also activated by the cytokines. These results suggest an integrative role for IL6 in the fine-tuning of the chromaffin cell response to a wide range of physiological and paraphysiological stressors, particularly when immune and endocrine stimuli converge.