Analytical validation of a semi-automated methodology for quantitative measurement of SARS-CoV-2 RNA in wastewater collected in northern New England.

Wastewater-based epidemiology (WBE) can be used to monitor the community presence of infectious disease pathogens of public health concern such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Viral nucleic acid has been detected in the stool of SARS-CoV-2-infected individuals. Asymptomatic SARS-CoV-2 infections make community monitoring difficult without extensive and continuous population screening. In this study, we validated a procedure that includes manual pre-processing, automated SARS-CoV-2 RNA extraction and detection workflows using both reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) and reverse transcriptase droplet digital PCR (RT-ddPCR). Genomic RNA and calibration materials were used to create known concentrations of viral material to determine the linearity, accuracy, and precision of the wastewater extraction and SARS-CoV-2 RNA detection. Both RT-qPCR and RT-ddPCR perform similarly in all the validation experiments, with a limit of detection of 50 copies/mL. A wastewater sample from a care facility with a known outbreak was assessed for viral content in replicate, and we showed consistent results across both assays. Finally, in a 2-week survey of two New Hampshire cities, we assessed the suitability of our methods for daily surveillance. This paper describes the technical validation of a molecular assay that can be used for long-term monitoring of SARS-CoV-2 in wastewater as a potential tool for community surveillance to assist with public health efforts.IMPORTANCEThis paper describes the technical validation of a molecular assay that can be used for the long-term monitoring of SARS-CoV-2 in wastewater as a potential tool for community surveillance to assist with public health efforts.

Single-cell epigenomic dysregulation of Systemic Sclerosis fibroblasts via CREB1/EGR1 axis in self-assembled human skin equivalents.

Systemic sclerosis (SSc) is an autoimmune disease characterized by skin fibrosis, internal organ involvement and vascular dropout. We previously developed and phenotypically characterized an in vitro 3D skin-like tissue model of SSc, and now analyze the transcriptomic (scRNA-seq) and epigenetic (scATAC-seq) characteristics of this model at single-cell resolution. SSc 3D skin-like tissues were fabricated using autologous fibroblasts, macrophages, and plasma from SSc patients or healthy control (HC) donors. SSc tissues displayed increased dermal thickness and contractility, as well as increased α-SMA staining. Single-cell transcriptomic and epigenomic analyses identified keratinocytes, macrophages, and five populations of fibroblasts (labeled FB1 - 5). Notably, FB1 APOE-expressing fibroblasts were 12-fold enriched in SSc tissues and were characterized by high EGR1 motif accessibility. Pseudotime analysis suggests that FB1 fibroblasts differentiate from a TGF-ß1-responsive fibroblast population and ligand-receptor analysis indicates that the FB1 fibroblasts are active in macrophage crosstalk via soluble ligands including FGF2 and APP. These findings provide characterization of the 3D skin-like model at single cell resolution and establish that it recapitulates subsets of fibroblasts and macrophage phenotypes observed in skin biopsies.

[Level of knowledge, attitudes and the use of preventive measures among household contacts of COVID-19 cases after the acute phase of the pandemic].

BACKGROUND: Assessment of the level of knowledge and attitudes regarding COVID-19 and preventive measures in household contacts of COVID-19 cases after the acute phase of the pandemic. METHODS: Survey among household contacts of COVID-19 cases conducted in healthcare centers in Navarre (1) and Catalonia (8) between May 2022 and July 2023. The frequency of use of preventive measures and the knowledge and attitudes towards COVID-19 were assessed through 12 items. RESULTS: 215 contacts participated who correctly used the preventive measures (>85%), except for facemasks (35.8%) and interpersonal distance (47%); >85% showed adequate knowledge (5/6 items) and >80% had a positive attitude (3/6 items). Moreover, 54.7% considered that COVID-19 negatively affected their life and 54.1% that it is better to develop immunity by getting infected than by vaccination. CONCLUSIONS: Household contacts show a correct level of knowledge and positive attitude towards the disease and its preventive measures.

Absence of short-term changes in knowledge and attitudes among household contacts of COVID-19 cases during the post-acute phase of the pandemic in Catalonia and Navarre, Spain.

Objectives: To evaluate short-term changes in knowledge and attitude towards COVID-19 and preventive measures during the post-acute phase of the pandemic in Spain. Methods: A survey was performed in Catalonia and Navarre between May-2022 and July-2023 in household contacts of COVID-19 cases. Knowledge and attitude were assessed at baseline and at three months, using a Likert scale. Responses were grouped according to correct or incorrect. Results: At baseline, 172 subjects were contacted, 118 (69%) of which completed a follow-up interview three months later. Knowledge of correct hand-washing and mask protocols had maintained over time (-1.7%, p = 0.553 and - 2.5%, p = 0.473, respectively). Attitudes toward preventive measures was adequate in the first interview (86%), but attitudes regarding use of face masks decreased significantly (-9.1%, p = 0.048) over time in participants with higher risk of severe COVID-19. However, most short-term changes in knowledge and attitudes were not statistically significant. Conclusion: Household contacts showed correct knowledge and attitude towards COVID-19 and its preventive measures, without significant changes in the short term despite a relaxation of government-mandated preventive measures. These results provide relevant information in case of a new health emergency due to respiratory viruses.

Vaccinated COVID-19 Index Cases Are Less Likely to Transmit SARS-CoV-2 to Their Household Contacts: A Cohort Study.

The aim of this study was to evaluate the impact of index case vaccination on SARS-CoV-2 transmission to household contacts. In our epidemiological cohort study (May 2022-November 2023), we surveyed registered index case vaccination status and test results for contacts (testing on day 0, and on day 7 for negative contacts) and calculated the secondary attack rate (SAR), i.e., newly infected contacts/susceptible included contacts. The association of the independent variable, index case COVID-19 vaccination (yes/no), with household contact infection was determined using the adjusted odds ratio (aOR) and its 95% confidence interval (CI). We recorded 181 index cases and 314 contacts, of whom 250 agreed to participate; 16 contacts were excluded upon testing positive on day 0. Of the 234 included contacts, 49.1% were women, and the mean (SD) age was 51.9 (19.8) years. The overall SAR of 37.2% (87/234) was lower in the contacts of both vaccinated index cases (34.9% vs. 63.2%; p = 0.014) and index cases with a previous SARS-CoV-2 infection history (27.0% vs. 46.3%; p = 0.002). Index case vaccination showed a protective effect against infection for their household contacts (aOR = 0.21; 95% CI: 0.07, 0.67). The household SAR was high when the Omicron variant circulated. Vaccinated index cases were less likely to transmit SARS-CoV-2 to their contacts.

Factors Associated with Non-Adherence to Tuberculosis Preventive Treatment among Adult Contacts of Pulmonary Tuberculosis Cases with Latent Tuberculosis Infection in Catalonia, Spain, in 2019-2021.

The aim of this study was to identify factors associated with non-adherence to tuberculosis (TB) preventive treatment among contacts with latent TB infection for new cases of pulmonary TB cases reported in Catalonia in 2019-2021. All contacts aged 18 years or older with a latent TB infection who received a TB preventive treatment were included in the study. The Chi square test and the odds ratios (OR) were used to assess the association between non-adherence to TB preventive treatment and the study variables; a p < 0.05 was considered statistically significant. Multiple logistic regression analysis was used to detect the independent factors associated with TB preventive treatment non-adherence; a p < 0.05 was considered statistically significant. The percentage of non-adherence to TB preventive treatment found in this study was 23.7%. A multivariable logistic regression analysis determined that the following factors were significantly associated with TB preventive treatment non-adherence among adult contacts: "exposure at school or workplace" (aOR = 3.34), "exposure to an index case without laboratory confirmation of TB" (aOR = 2.07), "immigrant contact" (aOR = 1.81), "male gender" (aOR = 1.75) and "exposure duration < 6 h per week or sporadic" (aOR = 1.60. By contrast, the factor "short-term TB preventive treatment regimen" (aOR = 0.38) was significantly associated with a lower treatment non-adherence. Adherence to TB preventive treatment should be improved among adult contacts of TB pulmonary cases with latent TB infection by recommending short-term treatment regimens and by developing health education activities, with a greater focus on contacts with factors associated with treatment non-adherence.

The Vaccine Training Barometer: Assessing healthcare providers' confidence to answer vaccine-related questions and their training needs.

Healthcare providers (HCP) are seen by the public as the most trustworthy source of information about vaccination. While HCPs could be a valuable partner to increase vaccine confidence in general, it is not clear whether they feel confident themselves to address questions concerning vaccination. In the context of the EU Joint Action on Vaccination (EU-JAV), the Vaccine Training Barometer, an online survey tool, was developed to assess how frequently HCPs receive questions about vaccination, how confident they feel to answer these questions, and to what extent they are willing to follow extra training. After a pilot test in Flanders, Belgium, the Barometer was launched and completed by 833 HCPs in Flanders and 291 HCPs in the Spanish regions of Catalonia, Navarre and Valencian Community from November 2020 until January 2021, during the COVID-19 pandemic, just before and during the start of the first COVID-19 vaccination campaigns. In both countries, HCPs frequently received questions about vaccination (mostly on a daily or weekly basis), and about two thirds of them indicated that the frequency of questions had increased during the three months prior to completing the survey. Most questions were about the side effects and safety of vaccines. In both countries, a considerable proportion of HCPs did not feel confident to answer vaccine-related questions (31.5% felt confident in Flanders, 21.6% in Spain). A large proportion of HCPs received questions in the last three months before the survey that they could not answer (52.4% of respondents in Flemish sample, 41.5% in Spanish sample). Only 11.4% (Flanders) and 11.3% (Spain) of the respondents felt they gained sufficient knowledge through their standard education to be able to answer questions about vaccination. Almost all respondents were willing to follow extra training on vaccination (Flanders: 95.4%, Spain: 96.6%). The Vaccine Training Barometer is thus a useful tool to monitor HCPs' confidence to answer questions about vaccination and to capture their training needs.

Nivel de conocimientos, de actitudes y utilización de medidas preventivas entre los contactos domiciliarios de casos de COVID-19 tras la fase aguda de la pandemia / Level of knowledge, attitudes and the use of preventive measures among household contacts of COVID-19 cases after the acute phase of the pandemic

Fundamento. Describir el nivel de conocimiento y actitudes sobre la COVID-19 y sus medidas preventivas en contactos domiciliarios de casos de COVID-19 tras la fase aguda de la pandemia. Métodos. Encuesta a contactos domiciliarios de casos de COVID-19 realizada en centros de salud de Navarra (1) y Cataluña (8) entre mayo/2022 y julio/2023. Se evaluó el uso de medidas preventivas y, mediante 12 ítems, los conocimientos y actitudes frente a COVID-19. Resultados. Participaron 215 contactos que usaron correctamente las medidas preventivas (>85%), exceptuando mascarilla (35,8%) y distancia interpersonal (47%); >85% mostraron conocimientos adecuados en 5/6 ítems y >80% mostraron actitud positiva en 3/6 ítems. El 54,7% consideró que la COVID-19 influyó negativamente en su vida y el 54,1% que es mejor desarrollar inmunidad enfermando que mediante la vacunación. Conclusiones. Los contactos domiciliarios mostraron un correcto nivel de conocimiento y una actitud positiva frente a la COVID-19 y sus medidas preventivas. (AU)

Background. Assessment of the level of knowledge and attitudes regarding COVID-19 and preventive measures in household contacts of COVID-19 cases after the acute phase of the pandemic. Methods. Survey among household contacts of COVID-19 cases conducted in healthcare centers in Navarre (1) and Catalonia (8) between May 2022 and July 2023. The frequency of use of preventive measures and the knowledge and attitudes towards COVID-19 were assessed through 12 items. Results. 215 contacts participated who correctly used the preventive measures (>85%), except for facemasks (35.8%) and interpersonal distance (47%); >85% showed adequate knowledge (5/6 items) and >80% had a positive attitude (3/6 items). Moreover, 54.7% considered that COVID-19 negatively affected their life and 54.1% that it is better to develop immunity by getting infected than by vaccination. Conclusions. Household contacts show a correct level of knowledge and positive attitude towards the disease and its preventive measures. (AU)

The frequency of pathogenic variation in the All of Us cohort reveals ancestry-driven disparities.

Disparities in data underlying clinical genomic interpretation is an acknowledged problem, but there is a paucity of data demonstrating it. The All of Us Research Program is collecting data including whole-genome sequences, health records, and surveys for at least a million participants with diverse ancestry and access to healthcare, representing one of the largest biomedical research repositories of its kind. Here, we examine pathogenic and likely pathogenic variants that were identified in the All of Us cohort. The European ancestry subgroup showed the highest overall rate of pathogenic variation, with 2.26% of participants having a pathogenic variant. Other ancestry groups had lower rates of pathogenic variation, including 1.62% for the African ancestry group and 1.32% in the Latino/Admixed American ancestry group. Pathogenic variants were most frequently observed in genes related to Breast/Ovarian Cancer or Hypercholesterolemia. Variant frequencies in many genes were consistent with the data from the public gnomAD database, with some notable exceptions resolved using gnomAD subsets. Differences in pathogenic variant frequency observed between ancestral groups generally indicate biases of ascertainment of knowledge about those variants, but some deviations may be indicative of differences in disease prevalence. This work will allow targeted precision medicine efforts at revealed disparities.

Risk of tuberculosis among pulmonary tuberculosis contacts: the importance of time of exposure to index cases.

INTRODUCTION: The intensity of exposure to index cases of tuberculosis [TB] may increase the risk of TB in their contacts. The aim was to determine TB risk factors among contacts of TB index cases. METHODS: A cohort study was carried out in the contacts of pulmonary TB cases registered by the epidemiological surveillance network from 01/01/2019 to 06/30/2021. The factors associated with the risk of TB in contacts were determined using the adjusted odds ratio [aOR] and its 95% confidence interval [CI]. RESULTS: From 847 TB cases, 7087 contacts were identified. The prevalence of TB was 2.0% [145/7087] and was higher in < 5 years compared to those ≥ 65 years [4.4% versus 1.2%; p < 0.001], in those exposed ≥ 6 h daily [4%], and < 6 h daily [1.6%] with respect to weekly exposure of < 6 h [0.7%; p < 0.001]. Those contacts exposed ≥ 6 h daily [aOR= 6.9; 95%CI:2.1-22.1], < 5 years [aOR= 8.3; 95%CI:1.8-37.8] and immigrants [aOR= 1.7; 95%CI:1.1-2.7] had a higher risk of TB. CONCLUSIONS: The risk of TB increases with the time of exposure to the index case and this risk is also higher in < 5 years and immigrants. Contact tracing has a high yield for detecting new cases of TB.

Factors Associated with Treatment Prescription to Pulmonary Tuberculosis Contacts in Catalonia (2019-2021): A Population-Based Epidemiological Study.

In countries with low tuberculosis (TB) incidence, the systematic testing and treatment of latent TB infection (LTBI) in contacts of pulmonary TB index cases is the standard of care. The objective of this study, conducted in Catalonia over 2019-2021, was to assess the factors associated with LTBI treatment prescription to close contacts of pulmonary TB index cases. In this population-based epidemiological study of LTBI prevalence among pulmonary TB contacts between 2019 and 2021, multiple logistic backward stepwise regression was used to identify the factors associated with treatment prescription, for which the adjusted odds ratio (aOR) and 95% confidence intervals (CI) were calculated. A total of 1487 LTBI contacts of 542 pulmonary TB index cases were studied, 80.6% of whom received a prescription. The factors associated with LTBI treatment prescription were exposure ≥6 h/day (aOR 14.20; 95% CI 5.22-38.66) and exposure <6 h/day (aOR 7.32, 95% CI 2.48-21.64), whereas the factors associated with no LTBI treatment prescription were age ≥55 years (aOR 0.22, 95% CI 0.08-0.64) and bacillus Calmette-Guerin vaccination (aOR 0.38, 95% CI 0.16-0.90). Crucial to LTBI treatment prescription is information on the contact's duration of exposure to pulmonary TB, not only for contacts exposed for ≥6 h/day, but also for contacts with lower daily exposure levels.

A Profile of Adult Severe Acute Respiratory Syndrome Coronavirus 2 Pneumonia Patients According to Pneumococcal Vaccination Status.

Certain patient profile characteristics, such as preexisting medical conditions, can modify the risk of developing SARS-CoV-2 pneumonia among adults vaccinated and not vaccinated against pneumococcal disease. This retrospective cohort study aimed to quantify the risk of pneumonia caused by SARS-CoV-2 among individuals from 15 to 64 years old with and without pneumococcal vaccination in Spain during the 2020-2021 influenza season and establish a risk profile of patients more likely to develop SARS-CoV-2 pneumonia. Data (demographic information, patient medical history, and lifestyle habits) were gathered both directly from the patient via personal interview and by reviewing electronic medical records. In an adjusted analysis for pneumococcal vaccinated patients, visits to hospital outpatient clinics were protective while visits to primary health care services, being widowed, obese, and not using masks in outdoor open spaces were identified as risk factors. For patients who had not received a pneumococcal vaccine, visits to hospital outpatient clinics were protective, while being overweight or obese, alcohol consumption, and not using masks in outdoor open spaces were identified as risk factors. Concerning comorbidities, in the pneumococcal vaccinated group none were found to be protective but having diabetes or other respiratory diseases were identified as risk factors. In the unvaccinated group, undergoing immunosuppressive treatment and having metastatic tumors were protective factors, while cerebrovascular disease and obesity with a BMI ≥ 40 were risk factors. A similar risk profile for developing SARS-CoV-2 pneumonia in pneumococcal vaccinated and non-vaccinated individuals was found. Generally, vaccinated individuals had a lower risk of developing SARS-CoV-2. The findings suggest that vaccination against S. pneumoniae could prevent and reduce SARS-CoV-2 pneumonia. Additionally, this study has identified individuals with other medical conditions, such as obesity, underweight, diabetes, and a history of respiratory diseases, who are at an increased risk of developing SARS-CoV-2 pneumonia and could benefit from vaccination and supervision.

Assessment of Knowledge and Attitudes Over Time in Postacute COVID-19 Environments: Protocol for an Epidemiological Study.

BACKGROUND: Globally, COVID-19 is in transition from the acute pandemic phase into a postacute phase, and special attention should be paid at this time to COVID-19 control strategies. Understanding public knowledge and attitudes plays a pivotal role in controlling COVID-19's spread and provides information about the public's adherence to preventive and control measures. OBJECTIVE: This study protocol describes the planning and management of a survey to investigate the persistent or changing trends in knowledge and attitudes regarding COVID-19, vaccination, and nonpharmaceutical preventive measures among COVID-19 cases' household contacts aged 18 years and older, after the acute phase of the pandemic in Catalonia and Navarre in Spain. The secondary objectives include investigating the rate of secondary transmission in households, taking into account the demographic characteristics, clinical manifestations, and preventive measures toward COVID-19. METHODS: A telephone questionnaire was designed to assess the changing trends in knowledge, preventive measures, and attitudes toward COVID-19 in 3 rounds (after identification as a household contact, 3 months later, and 6 months later). The questionnaire was developed following an extensive literature review and through discussions with a panel of experts who designed and assessed the validity of the questionnaire in terms of relevance, consistency, completeness, and clarity. The questionnaire consists of the following 7 sections: social and demographic characteristics (ie, gender, age, educational level, and workplace), comorbidities and risk factors (according to the recommendations from the COVID-19 vaccination strategy), epidemiological data (ie, exposure time, relationship with index cases, and frequency of use of nonpharmaceutical preventive measures), COVID-19 vaccination status (ie, the number and date of doses received), knowledge and attitudes toward COVID-19 (assessed using a 5-point Likert scale-totally agree, agree, neither agree nor disagree, disagree, and totally disagree), and sources of information (including traditional mass media, social media, and official sources). RESULTS: A pilot study was performed in May 2022 to evaluate the questionnaire with 22 household contacts. Preliminary findings indicated that the questionnaire was feasible and acceptable in the general population. The average response time was 15 minutes, with greater variations in responses by older participants. After the pilot study, recruitment of participants began and is expected to be completed at the end of the year 2023, after which the final results will be available in 2024. CONCLUSIONS: Despite the low transmission levels of SARS-CoV-2 and the relaxation of containment measures, the implementation of the survey during the postacute phase will provide valuable insight to assist public health decision-making and control the transmission of SARS-CoV-2 and other respiratory viruses, thereby attenuating the negative effects of COVID-19 at individual and population level. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/52114.

Risk of partial school closure for COVID-19 by socio-economic level in the period 2020-22.

BACKGROUND: The objective was to analyse the risk of partial school closure by economic level in Barcelona city. METHODS: In this ecological study, the risk of partial school closure for the academic years 2020-21 and 2021-22 was estimated by dividing the total number of days that each child was in quarantine or isolation by the total number of days that each child was at risk to be in quarantine or isolation in the academic year. The association between partial school closure risk and mean income by district was estimated with the Spearman rho. RESULTS: The lower the mean income, the higher the risk of partial closure (Spearman rho = 0.83; P-value = 0.003) during the academic year 2020-21. Specifically, the children from the district with the lowest income had a six times greater risk of partial school closure compared with those from the highest-income district. This risk did not show a significant socioeconomic gradient in the academic year 2021-22. CONCLUSIONS: The risk of partial school closure presented an inverse socioeconomic gradient in the city of Barcelona according to average income by district in the academic year 2020-21. This distribution was not observed in the academic year 2021-22.

Atherosclerosis, Cardiovascular Disease, and COVID-19: A Narrative Review.

Atherosclerosis is a chronic inflammatory and degenerative process that mainly occurs in large- and medium-sized arteries and is morphologically characterized by asymmetric focal thickenings of the innermost layer of the artery, the intima. This process is the basis of cardiovascular diseases (CVDs), the most common cause of death worldwide. Some studies suggest a bidirectional link between atherosclerosis and the consequent CVD with COVID-19. The aims of this narrative review are (1) to provide an overview of the most recent studies that point out a bidirectional relation between COVID-19 and atherosclerosis and (2) to summarize the impact of cardiovascular drugs on COVID-19 outcomes. A growing body of evidence shows that COVID-19 prognosis in individuals with CVD is worse compared with those without. Moreover, various studies have reported the emergence of newly diagnosed patients with CVD after COVID-19. The most common treatments for CVD may influence COVID-19 outcomes. Thus, their implication in the infection process is briefly discussed in this review. A better understanding of the link among atherosclerosis, CVD, and COVID-19 could proactively identify risk factors and, as a result, develop strategies to improve the prognosis for these patients.

Clinical Perspective on Use of Long-Read Sequencing in Prenatal Diagnosis of Thalassemia.

This is an editorial focusing on the clinical perspective of a long-read sequencing method in the prenatal diagnosis of alpha- and beta-thalassemia, including a comparison between this method and standard PCR-based methods. Though incremental, the increased sensitivity and specificity using long-read sequencing is an important advantage of this methodology in the prenatal diagnostic arena due to false positive or false negative results having greater consequence when a family is making decisions about their pregnancy.

Does knowing the influenza epidemic threshold has been reached influence the performance of influenza case definitions?

BACKGROUND: Disease surveillance using adequate case definitions is very important. The objective of the study was to compare the performance of influenza case definitions and influenza symptoms in the first two epidemic weeks with respect to other epidemic weeks. METHODS: We analysed cases of acute respiratory infection detected by the network of sentinel primary care physicians of Catalonia for 10 seasons. We calculated the diagnostic odds ratio (DOR) and 95% confidence intervals (CI) for the first two epidemic weeks and for other epidemic weeks. RESULTS: A total of 4,338 samples were collected in the epidemic weeks, of which 2,446 (56.4%) were positive for influenza. The most predictive case definition for laboratory-confirmed influenza was the WHO case definition for influenza-like illness (ILI) in the first two epidemic weeks (DOR 2.10; 95% CI 1.57-2.81) and in other epidemic weeks (DOR 2.31; 95% CI 1.96-2.72). The most predictive symptom was fever. After knowing that epidemic threshold had been reached, the DOR of the ILI WHO case definition in children aged <5 years and cough and fever in this group increased (190%, 170% and 213%, respectively). CONCLUSIONS: During influenza epidemics, differences in the performance of the case definition and the discriminative ability of symptoms were found according to whether it was known that the epidemic threshold had been reached or not. This suggests that sentinel physicians are stricter in selecting samples to send to the laboratory from patients who present symptoms more specific to influenza after rather than before an influenza epidemic has been declared.

A genomic meta-analysis of clinical variables and their association with intrinsic molecular subsets in systemic sclerosis.

OBJECTIVES: Four intrinsic molecular subsets (inflammatory, fibroproliferative, limited, normal-like) have previously been identified in SSc and are characterized by unique gene expression signatures and pathways. The intrinsic subsets have been linked to improvement with specific therapies. Here, we investigated associations between baseline demographics and intrinsic molecular subsets in a meta-analysis of published datasets. METHODS: Publicly available gene expression data from skin biopsies of 311 SSc patients measured by DNA microarray were classified into the intrinsic molecular subsets. RNA-sequencing data from 84 participants from the ASSET trial were used as a validation cohort. Baseline clinical demographics and intrinsic molecular subsets were tested for statistically significant associations. RESULTS: Males were more likely to be classified in the fibroproliferative subset (P = 0.0046). SSc patients who identified as African American/Black were 2.5 times more likely to be classified as fibroproliferative compared with White/Caucasian patients (P = 0.0378). ASSET participants sera positive for anti-RNA pol I and RNA pol III autoantibodies were enriched in the inflammatory subset (P = 5.8 × 10-5, P = 9.3 × 10-5, respectively), while anti-Scl-70 was enriched in the fibroproliferative subset. Mean modified Rodnan Skin Score (mRSS) was statistically higher in the inflammatory and fibroproliferative subsets compared with normal-like (P = 0.0027). The average disease duration for inflammatory subset was less than fibroproliferative and normal-like intrinsic subsets (P = 8.8 × 10-4). CONCLUSIONS: We identified multiple statistically significant differences in baseline demographics between the intrinsic subsets that may represent underlying features of disease pathogenesis (e.g. chronological stages of fibrosis) and have implications for treatments that are more likely to work in certain SSc populations.

Wastewater-Based SARS-CoV-2 Surveillance in Northern New England.

SARS-CoV-2 viral RNA is shed in the stool of 55-70% of infected individuals and can be detected in community wastewater up to 7 days before people present with COVID-19 symptoms. The detection of SARS-CoV-2 RNA in wastewater may serve as a lead indicator of increased community transmission. Here, we monitored viral concentrations in samples collected from nine municipal wastewater facilities in New Hampshire (NH) and Vermont (VT).Twenty-four-h composite primary influent wastewater samples were collected from nine municipal wastewater treatment facilities twice per week for 5 months (late September 2020 to early February 2021). Wastewater was centrifuged for 30 min at 4600 × g, then the supernatant was frozen until further analysis. Once thawed, samples were concentrated, extracted, and tested for SARS-CoV-2 RNA using reverse transcriptase-quantitative PCR (RT-qPCR) and reverse transcriptase-droplet digital PCR (RT-ddPCR) detection methods. Active case counts for each municipality were tracked from the NH and VT state COVID-19 dashboards. We received a total of 283 wastewater samples from all sites during the study period. Viral RNA was detected in 175/283 (61.8%) samples using RT-qPCR and in 195/283 (68.9%) samples using RT-ddPCR. All nine sites showed positivity in the wastewater, with 8/9 (88.8%) sites having over 50% of their samples test positive over the course of the study. Larger municipalities, such as Nashua, Concord, and Lebanon, NH, showed that SARS-CoV-2 positivity in the wastewater can precede spikes in active COVID-19 case counts by as much as 7 days. Smaller municipalities, such as Woodsville, NH and Hartford, VT, showed sporadic SARS-COV-2 detection and did not always precede a rise in active case counts. We detected SARS-CoV-2 RNA in samples from all 9 municipalities tested, including cities and small towns within this region, and showed wastewater positivity as an early indicator of active case count increases in some regions. Some of the smaller rural municipalities with low case counts may require more frequent sampling to detect SARS-CoV-2 in wastewater before a case surge. With timely collection and analysis of wastewater samples, a community could potentially respond to results by increasing public health initiatives, such as tightening mask mandates and banning large indoor gatherings, to mitigate community transmission of SARS-CoV-2. IMPORTANCE Despite vaccination efforts, the delta and omicron variants of SARS-CoV-2 have caused global surges of COVID-19. As the COVID-19 pandemic continues, it is important to find new ways of tracking early signs of SARS-CoV-2 outbreaks. The manuscript outlines how to collect wastewater from treatment facilities, concentrate the virus in a dilute wastewater sample, and detect it using two sensitive PCR-based methods. It also describes important trends in SARS-CoV-2 concentration in wastewater of a rural region of the United States from Fall 2020 - Winter 2021 and demonstrates the utility of wastewater monitoring as a leading indicator of active SARS-CoV-2 cases. Monitoring changes in concentration of SARS-CoV-2 virus in wastewater may offer an early indicator of increased case counts and enable appropriate public health actions to be taken.

Utility of a virtual counselor (VICKY) to collect family health histories among vulnerable patient populations: A randomized controlled trial.

OBJECTIVES: This study is a randomized controlled trial comparing the efficacy of a virtual counselor (VICKY) to the My Family Health Portrait (MFHP) tool for collecting family health history (FHx). METHODS: A total of 279 participants were recruited from a large safety-net hospital and block randomized by health literacy to use one of the digital FHx tools, followed by a genetic counselor interview. A final sample of 273 participants were included for analyses of primary study aims pertaining to tool concordance, which assessed agreement between tool and genetic counselor. RESULTS: Tool completion differed significantly between tools (VICKY = 97%, MFHP = 51%; p < .0001). Concordance between tool and genetic counselor was significantly greater for participants randomized to VICKY compared to MFHP for ascertaining first- and second-degree relatives (ps<.0001), and most health conditions examined. There was significant interaction by health literacy, with greater differences in concordance observed between tools among those with limited literacy. CONCLUSIONS: A virtual counselor overcomes many of the literacy-related barriers to using traditional digital tools and highlights an approach that may be important to consider when collecting health histories from vulnerable populations. PRACTICE IMPLICATIONS: The usability of digital health history tools will have important implications for the quality of the data collected and its downstream clinical utility.