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An outlook on urobiome: advances in understanding the role of urobiome in urological health and disease and its potential in biotherapeutics Kicking it off: current outlook on the human urobiome High-throughput, culture-independent technologies such as next-generation sequencing of the bacterial 16S ribosomal RNA (rRNA) gene, have become a phenomenal milestone in the exploration of the structure, function, and diversity of the microbiome of various body sites (1). Despite these recent strides, the bladder, urethra, and urinary microbiome (urobiome) remains relatively understudied; yet it may contribute to urological infections and diseases (2, 3), including urinary tract infections (UTIs), the most common outpatient infections (4). Available literature on urobiome exhibits heterogeneity in study methodologies, with a majority of studies having relatively small sample sizes and focusing on associations rather than causations. There are other fundamental questions that are yet to be examined. For instance, the origin, temporal stability, and individuality of the core urobiome, its dynamic and intricate interplay with the host immunity, urinary incontinence and various uropathologies, other factors (e.g., host genetics) impacting it, Frontiers in Urology frontiersin.org 01

Abstract BackgroundAgitation is a prevalent and disabling neuropsychiatric symptom of dementia that comprises a constellation of behaviours involving excessive motor activity, physical aggression, and verbal aggression. Pro re nata (PRN) injections of antipsychotics and benzodiazepines can be administered for severe acute agitation, but best practices surrounding their use remain poorly characterized. The aim of this study was to compare the frequency of use of injectable PRN haloperidol, olanzapine, and lorazepam for acute agitation in Canadian long‐term care (LTC) residents with dementia before and during the COVID‐19 pandemic.MethodsA retrospective chart review was conducted for patients from two Canadian LTC facilities who had orders for PRN haloperidol, olanzapine, or lorazepam between January 1, 2018–May 1, 2019 (i.e. pre‐COVID‐19) and January 1, 2020–May 1, 2021 (i.e. COVID‐19). We reviewed electronic medical records to document PRN administrations of these psychotropic medications and collect data on age, sex, dementia diagnosis, days of observation, care type, medical history, and concomitant medications. Descriptive statistics and multivariate logistic and Poisson regression models were used to analyze the data.ResultsOf the 250 residents with dementia included, 45 of 103 (44%) people in the pre‐COVID‐19 period and 85 of 147 (58%) people in the COVID‐19 period with standing orders for injectable PRN haloperidol, olanzapine, or lorazepam received ≥1 injections of their prescribed medication. Haloperidol was the most frequently used agent (74% pre‐COVID‐19 and 81% during COVID‐19) and excessive motor activity (85% and 80%) was the most common indication for use in both time periods. Residents in the COVID‐19 period were two times more likely to receive PRN injections compared with those in the pre‐COVID‐19 period (OR = 1.96; 95% CI = 1.15‐3.34; p = 0.01). Count of injections per patient was not significantly different between the two time periods (IRR = 1.35; 95% CI = 0.78‐2.33; p = 0.3).ConclusionOur results contribute to the mounting evidence that agitation worsened in LTC residents with dementia during the pandemic and add to the limited existing literature on actual use of PRN psychotropic medications for acute agitation. The increased need for PRN rescue medications may be an indirect consequence of COVID‐19 restrictions, particularly the restrictions on socialization.

Background The vast majority of SARS-CoV-2 vaccines which are licensed or under development focus on the spike (S) protein and its receptor binding domain (RBD). However, S and RBD from SARS-CoV-2 variants of concerns show considerable sequence variations and repeated injections for boosting specific immunity are necessary. Aim of this study was to develop and characterize a SARS-CoV-2 vaccine targeting the highly conserved nucleocapsid (N) protein. Methods Recombinant N protein was expressed in Escherichia coli, purified to homogeneity by chromatography and characterized by SDS-PAGE, immunoblotting, mass spectrometry, dynamic light scattering and differential scanning calorimetry. The N protein vaccine was obtained by formulation of recombinant N as squalane-based emulsion and used to immunize Balb/c mice, NOD scid gamma (NSG) mice engrafted with human PBMC, rabbits and marmoset monkeys to study safety as well as antibody and cellular immunity using ELISA for antibodies, measurement of N-specific Th1 and Th2 cytokine secretion and carboxyfluorescein succinimidyl ester (CFSE) dilution assays for CD4 and CD8 T cell responses. The protective effect of the vaccine was studied in SARS-CoV-2-infected Syrian hamsters. Results Immunization of mice, rabbits and Syrian hamsters with the recombinant N protein-based vaccine formulated as squalane-based emulsion (Convacell®) induced sustainable N-specific IgG responses and a N-specific mixed Th1/Th2 cytokine response. In marmoset monkeys a N-specific CD4 as well as CD8 T cell response was observed. Vaccinated and then infected Syrian hamsters showed reduced lung histopathology, reduced virus was detected in lung tissue, lung weight relative to the body was not increased after challenge and body weight was regained faster than in non-vaccinated animals. Repeated dose toxicity studies in mice and rabbits showed that Convacell® was well tolerated and safe. Conclusions Convacell® induced a SARS-CoV-2-specific protective immune response in Syrian hamsters. It is a new vaccine targeting the nucleocapsid protein of SARS-CoV-2 and thus may augment the armamentarium of vaccines for COVID-19.

ObjectiveAs SARS‐CoV‐2 is known to invade neural cell mitochondria, a plasma system for quantifying central nervous system proteins in living humans was used to investigate neuropathogenic mechanisms of long‐COVID‐19.MethodsSARS‐CoV‐2 proteins and mitochondrial proteins (MPs) in enriched plasma neuron‐derived extracellular vesicles (NDEVs) and astrocyte‐derived EVs (ADEVs) were quantified in resolved acute COVID‐19 without post‐acute sequelae of SARS‐CoV‐2 (PASC), PASC without neuropsychiatric manifestations (NP), PASC with NP and healthy controls.ResultsNDEV and ADEV mean levels of SARS‐CoV‐2 S1 and nucleocapsid (N) proteins were higher in all PASC sub‐groups than controls, but only N levels were higher in PASC with than without NP. Exosome marker CD81‐normalized NDEV mean levels of subunit 6 of MP respiratory chain complex I and subunit 10 of complex III, and neuroprotective MPs Humanin and mitochondrial open‐reading frame of the 12S rRNA‐c (MOTS‐c) all were decreased significantly in PASC with NP but not in PASC without NP relative to controls. NDEV levels of MPs voltage‐dependent anion‐selective channel protein 1 (VDAC1) and N‐methyl‐D‐aspartate receptor 1 (NMDAR1) were decreased in PASC without and with NP, whereas those of calcium channel MPs mitochondrial calcium uniporter (MCU), sodium/calcium exchanger (NCLX) and leucine zipper EF‐hand containing transmembrane 1 protein (LETM1) were decreased only in PASC with NP. ADEV levels of MCU and NCLX only were increased in PASC without and with NP.InterpretationAbnormal NDEV and ADEV levels of SARS‐CoV‐2 N and S1 protein and MPs correlate with NP and may be biomarkers for long‐COVID prognostics and therapeutic trials. ANN NEUROL 2022;91:772–781