Proteomic Analysis of Mucosal and Systemic Responses to SARS-CoV-2 Antigen.

The mucosal environment of the upper respiratory tract is the first barrier of protection against SARS-CoV-2 transmission. However, the mucosal factors involved in viral transmission and potentially modulating the capacity to prevent such transmission have not fully been identified. In this pilot proteomics study, we compared mucosal and systemic compartments in a South African cohort of vaccinated and unvaccinated individuals undergoing maxillofacial surgery with previous history of COVID-19 or not. Inflammatory profiles were analyzed in plasma, nasopharyngeal swabs, and nasal and oral tissue explant cultures, using Olink and Luminex technologies. SARS-CoV-2-specific antibody levels were measured in serum and tissue explants. An increased pro-inflammatory proteomic profile was measured in the nasal compartment compared to plasma. However, IP-10 and MIG levels were higher in secretions than in nasal tissue, and the opposite was observed for TGF-ß. Nasal anti-SARS-CoV-2 spike IgG correlated with mucosal MIG expression for all participants. A further positive correlation was found with IP-10 in BioNTech/Pfizer-vaccinated individuals. Systemic levels of anti-SARS-CoV-2 spike IgG elicited by this vaccine correlated with plasma IL-10, IL-6 and HBD4. Proteomic profiles measured in mucosal tissues and secretions using combined technologies could reveal correlates of protection at the mucosal portals of viral entry.

Locked down or locked out? Trends in psychiatric emergency services utilization during the COVID-19 pandemic.

OBJECTIVE: To estimate changes in Boston Emergency Services Team (BEST) psychiatric emergency services (PES) encounter volume (total and by care team) and inpatient disposition during the first 8 months of the COVID-19 pandemic. METHODS: Data on 30,657 PES encounters was extracted from the four-county, BEST reporting system. The study period consisted of the first 34 weeks of 2019 and 2020. This period corresponded to the first five stages of Massachusetts's COVID-19 public health restrictions: pre-lockdown, lockdown, Phase I, II and III reopenings. Descriptive and regression analyses were performed to estimate changes in encounter volume by care team and disposition. RESULTS: Compared to the same period in 2019, covariate-adjusted, weekly PES encounters decreased by 39% (ß = -0.40, 95% Confidence Interval (CI) = [-0.51, -0.28], p < 0.00) during the lockdown. PES volume remained significantly lower during Phase I reopening compared to the previous year but returned to 2019 levels during Phase II. The covariate-adjusted proportion of weekly encounters that led to inpatient admission significantly increased by 16% (CI = [0.11, 0.21], p < 0.00) for mobile crisis teams (MCTs) and significantly declined by 13% (CI = [-0.19, -0.07], p < 0.00) for BEST-designated emergency departments during the lockdown period compared to the prior year. CONCLUSIONS: The overall drop in PES utilization and the rise in inpatient admissions for MCT encounters suggests that during the early phases of the pandemic, patients delayed psychiatric care until they had a psychiatric crisis. Public health messaging about the lockdowns and absent equivalent messaging about the availability of telehealth services may have made patients more reluctant to seek psychiatric care.

Psychiatric emergencies among urban youth during COVID-19: Volume and acuity in a multi-channel program for the publicly insured

This study sought to characterize changes in the utilization of psychiatric emergency services among children and adolescents during distinct phases of 2020, as compared with prior years. We conducted a retrospective review of electronic health records from January 2018 through December 2020 that included all encounters made by patients under age 21. We then analyzed data for the 15,045 youth psychiatric encounters during the study period. Encounter volume in 2020 was significantly lower than prior years in March through May (IRR, 0.44;95% CI, 0.40–0.49), May through July (IRR, 0.63;95% CI, 0.56–0.71), and October through December (IRR, 0.76;95% CI, 0.70–0.83). Encounters for youth with primary psychotic disorders remained at typical levels throughout 2020. Among older adolescents and youth with anxiety disorders, pervasive developmental disorders, and substance use disorders, encounter volume was significantly lower than prior years only during the initial lockdown period. There were significantly more encounters than normal conducted by mobile crisis units, including via telehealth, in July through October (IRR, 1.31;95% CI, 1.06–1.62) and October through December (IRR, 1.28;95% CI, 1.05–1.55) of 2020. Differences in patterns of encounter volume based on sociodemographic and clinical characteristics highlight subgroups of youth who may have been particularly vulnerable to acute mental health problems during periods of social distancing and isolation. Proactive efforts to engage vulnerable youth in outpatient treatment during periods of increased infectivity may help prevent increasing symptoms from reaching the point of crisis.

Psychiatric emergencies among urban youth during COVID-19: Volume and acuity in a multi-channel program for the publicly insured.

This study sought to characterize changes in the utilization of psychiatric emergency services among children and adolescents during distinct phases of 2020, as compared with prior years. We conducted a retrospective review of electronic health records from January 2018 through December 2020 that included all encounters made by patients under age 21. We then analyzed data for the 15,045 youth psychiatric encounters during the study period. Encounter volume in 2020 was significantly lower than prior years in March through May (IRR, 0.44; 95% CI, 0.40-0.49), May through July (IRR, 0.63; 95% CI, 0.56-0.71), and October through December (IRR, 0.76; 95% CI, 0.70-0.83). Encounters for youth with primary psychotic disorders remained at typical levels throughout 2020. Among older adolescents and youth with anxiety disorders, pervasive developmental disorders, and substance use disorders, encounter volume was significantly lower than prior years only during the initial lockdown period. There were significantly more encounters than normal conducted by mobile crisis units, including via telehealth, in July through October (IRR, 1.31; 95% CI, 1.06-1.62) and October through December (IRR, 1.28; 95% CI, 1.05-1.55) of 2020. Differences in patterns of encounter volume based on sociodemographic and clinical characteristics highlight subgroups of youth who may have been particularly vulnerable to acute mental health problems during periods of social distancing and isolation. Proactive efforts to engage vulnerable youth in outpatient treatment during periods of increased infectivity may help prevent increasing symptoms from reaching the point of crisis.

Assessing a novel, lab-free, point-of-care test for SARS-CoV-2 (CovidNudge): a diagnostic accuracy study.

BACKGROUND: Access to rapid diagnosis is key to the control and management of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Laboratory RT-PCR testing is the current standard of care but usually requires a centralised laboratory and significant infrastructure. We describe our diagnostic accuracy assessment of a novel, rapid point-of-care real time RT-PCR CovidNudge test, which requires no laboratory handling or sample pre-processing. METHODS: Between April and May, 2020, we obtained two nasopharyngeal swab samples from individuals in three hospitals in London and Oxford (UK). Samples were collected from three groups: self-referred health-care workers with suspected COVID-19; patients attending emergency departments with suspected COVID-19; and hospital inpatient admissions with or without suspected COVID-19. For the CovidNudge test, nasopharyngeal swabs were inserted directly into a cartridge which contains all reagents and components required for RT-PCR reactions, including multiple technical replicates of seven SARS-CoV-2 gene targets (rdrp1, rdrp2, e-gene, n-gene, n1, n2 and n3) and human ribonuclease P (RNaseP) as sample adequacy control. Swab samples were tested in parallel using the CovidNudge platform, and with standard laboratory RT-PCR using swabs in viral transport medium for processing in a central laboratory. The primary analysis was to compare the sensitivity and specificity of the point-of-care CovidNudge test with laboratory-based testing. FINDINGS: We obtained 386 paired samples: 280 (73%) from self-referred health-care workers, 15 (4%) from patients in the emergency department, and 91 (23%) hospital inpatient admissions. Of the 386 paired samples, 67 tested positive on the CovidNudge point-of-care platform and 71 with standard laboratory RT-PCR. The overall sensitivity of the point-of-care test compared with laboratory-based testing was 94% (95% CI 86-98) with an overall specificity of 100% (99-100). The sensitivity of the test varied by group (self-referred healthcare workers 94% [95% CI 85-98]; patients in the emergency department 100% [48-100]; and hospital inpatient admissions 100% [29-100]). Specificity was consistent between groups (self-referred health-care workers 100% [95% CI 98-100]; patients in the emergency department 100% [69-100]; and hospital inpatient admissions 100% [96-100]). Point of care testing performance was similar during a period of high background prevalence of laboratory positive tests (25% [95% 20-31] in April, 2020) and low prevalence (3% [95% 1-9] in inpatient screening). Amplification of viral nucleocapsid (n1, n2, and n3) and envelope protein gene (e-gene) were most sensitive for detection of spiked SARS-CoV-2 RNA. INTERPRETATION: The CovidNudge platform was a sensitive, specific, and rapid point of care test for the presence of SARS-CoV-2 without laboratory handling or sample pre-processing. The device, which has been implemented in UK hospitals since May, 2020, could enable rapid decisions for clinical care and testing programmes. FUNDING: National Institute of Health Research (NIHR) Imperial Biomedical Research Centre, NIHR Health Protection Research Unit in Healthcare Associated Infections and Antimicrobial Resistance at Oxford University in partnership with Public Health England, NIHR Biomedical Research Centre Oxford, and DnaNudge.

Optimized protocol for a quantitative SARS-CoV-2 duplex RT-qPCR assay with internal human sample sufficiency control.

There is growing evidence that measurement of SARS-CoV-2 viral copy number can inform clinical and public health management of SARS-CoV-2 carriers and COVID-19 patients. Here we show that quantification of SARS-CoV-2 is feasible in a clinical setting, using a duplex RT-qPCR assay which targets both the E gene (Charité assay) and a human RNA transcript, RNase P (CDC assay) as an internal sample sufficiency control. Samples in which RNase P is not amplified indicate that sample degradation has occurred, PCR inhibitors are present, RNA extraction has failed or swabbing technique was insufficient. This important internal control reveals that 2.4 % of nasopharyngeal swabs (15/618 samples) are inadequate for SARS-CoV-2 testing which, if not identified, could result in false negative results. We show that our assay is linear across at least 7 logs and is highly reproducible, enabling the conversion of Cq values to viral copy numbers using a standard curve. Furthermore, the SARS-CoV-2 copy number was independent of the RNase P copy number indicating that the per-swab viral copy number is not dependent on sampling- further allowing comparisons between samples. The ability to quantify SARS-CoV-2 viral copy number will provide an important opportunity for viral burden-guided public health and clinical decision making.

Kinin B1 Receptor Is Important in the Pathogenesis of Myeloperoxidase-Specific ANCA GN.

BACKGROUND: Myeloperoxidase-specific ANCA (MPO-ANCA) are implicated in the pathogenesis of vasculitis and GN. Kinins play a major role during acute inflammation by regulating vasodilatation and vascular permeability and by modulating adhesion and migration of leukocytes. Kinin system activation occurs in patients with ANCA vasculitis. Previous studies in animal models of GN and sclerosing kidney diseases have demonstrated protective effects of bradykinin receptor 1 (B1R) blockade via interference with myeloid cell trafficking. METHODS: To investigate the role of B1R in a murine model of MPO-ANCA GN, we evaluated effects of B1R genetic ablation and pharmacologic blockade. We used bone marrow chimeric mice to determine the role of B1R in bone marrow-derived cells (leukocytes) versus nonbone marrow-derived cells. We elucidated mechanisms of B1R effects using in vitro assays for MPO-ANCA-induced neutrophil activation, endothelial adherence, endothelial transmigration, and neutrophil adhesion molecule surface display. RESULTS: B1R deficiency or blockade prevented or markedly reduced ANCA-induced glomerular crescents, necrosis, and leukocyte influx in mice. B1R was not required for in vitro MPO-ANCA-induced neutrophil activation. Leukocyte B1R deficiency, but not endothelial B1R deficiency, decreased glomerular neutrophil infiltration induced by MPO-ANCA in vivo. B1R enhanced ANCA-induced neutrophil endothelial adhesion and transmigration in vitro. ANCA-activated neutrophils exhibited changes in Mac-1 and LFA-1, important regulators of neutrophil endothelial adhesion and transmigration: ANCA-activated neutrophils increased surface expression of Mac-1 and increased shedding of LFA-1, whereas B1R blockade reduced these effects. CONCLUSIONS: The leukocyte B1R plays a critical role in the pathogenesis of MPO-ANCA-induced GN in a mouse model by modulating neutrophil-endothelial interaction. B1R blockade may have potential as a therapy for ANCA GN and vasculitis.