The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish

The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 μM was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood.info:eu-repo/semantics/publishedVersio

Using a state cancer registry to recruit young breast cancer survivors and high-risk relatives: protocol of a randomized trial testing the efficacy of a targeted versus a tailored intervention to increase breast cancer screening

Abstract Background The Michigan Prevention Research Center, the University of Michigan Schools of Nursing, Public Health, and Medicine, and the Michigan Department of Community Health propose a multidisciplinary academic-clinical practice three-year project to increase breast cancer screening among young breast cancer survivors and their cancer-free female relatives at greatest risk for breast cancer. Methods/design The study has three specific aims: 1) Identify and survey 3,000 young breast cancer survivors (diagnosed at 20–45 years old) regarding their breast cancer screening utilization. 2) Identify and survey survivors’ high-risk relatives regarding their breast cancer screening utilization. 3) Test two versions (Targeted vs. Enhanced Tailored) of an intervention to increase breast cancer screening among survivors and relatives. Following approval by human subjects review boards, 3,000 young breast cancer survivors will be identified through the Michigan Cancer Registry and mailed an invitation letter and a baseline survey. The baseline survey will obtain information on the survivors’: a) current breast cancer screening status and use of genetic counseling; b) perceived barriers and facilitators to screening; c) family health history. Based on the family history information provided by survivors, we will identify up to two high-risk relatives per survivor. Young breast cancer survivors will be mailed consent forms and baseline surveys to distribute to their selected high-risk relatives. Relatives’ baseline survey will obtain information on their: a) current breast cancer screening status and use of genetic counseling; and b) perceived barriers and facilitators to screening. Young breast cancer survivors and high-risk relatives will be randomized as a family unit to receive two versions of an intervention aiming to increase breast cancer screening and use of cancer genetic services. A follow-up survey will be mailed 9 months after the intervention to survivors and high-risk relatives to evaluate the efficacy of each intervention version on: a) use of breast cancer screening and genetic counseling; b) perceived barriers and facilitators to screening; c) self-efficacy in utilizing cancer genetic and screening services; d) family support related to screening; e) knowledge of breast cancer genetics; and f) satisfaction with the intervention. Discussion The study will enhance efforts of the state of Michigan surrounding cancer prevention, control, and public health genomics. Trial registration NCT01612338http://deepblue.lib.umich.edu/bitstream/2027.42/112835/1/12885_2012_Article_3739.pd

The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish

The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 μM was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood.info:eu-repo/semantics/publishedVersio

Single-cell meta-analysis of SARS-CoV-2 entry genes across tissues and demographics

Angiotensin-converting enzyme 2 (ACE2) and accessory proteases (TMPRSS2 and CTSL) are needed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) cellular entry, and their expression may shed light on viral tropism and impact across the body. We assessed the cell-type-specific expression of ACE2, TMPRSS2 and CTSL across 107 single-cell RNA-sequencing studies from different tissues. ACE2, TMPRSS2 and CTSL are coexpressed in specific subsets of respiratory epithelial cells in the nasal passages, airways and alveoli, and in cells from other organs associated with coronavirus disease 2019 (COVID-19) transmission or pathology. We performed a meta-analysis of 31 lung single-cell RNA-sequencing studies with 1,320,896 cells from 377 nasal, airway and lung parenchyma samples from 228 individuals. This revealed cell-type-specific associations of age, sex and smoking with expression levels of ACE2, TMPRSS2 and CTSL. Expression of entry factors increased with age and in males, including in airway secretory cells and alveolar type 2 cells. Expression programs shared by ACE2+TMPRSS2+ cells in nasal, lung and gut tissues included genes that may mediate viral entry, key immune functions and epithelial-macrophage cross-talk, such as genes involved in the interleukin-6, interleukin-1, tumor necrosis factor and complement pathways. Cell-type-specific expression patterns may contribute to the pathogenesis of COVID-19, and our work highlights putative molecular pathways for therapeutic intervention

Associations of Underlying Health Conditions With Anxiety and Depression Among Outpatients: Modification Effects of Suspected COVID-19 Symptoms, Health-Related and Preventive Behaviors

Objectives: We explored the association of underlying health conditions (UHC) with depression and anxiety, and examined the modification effects of suspected COVID-19 symptoms (S-COVID-19-S), health-related behaviors (HB), and preventive behaviors (PB).Methods: A cross-sectional study was conducted on 8,291 outpatients aged 18–85 years, in 18 hospitals and health centers across Vietnam from 14th February to May 31, 2020. We collected the data regarding participant's characteristics, UHC, HB, PB, depression, and anxiety.Results: People with UHC had higher odds of depression (OR = 2.11; p < 0.001) and anxiety (OR = 2.86; p < 0.001) than those without UHC. The odds of depression and anxiety were significantly higher for those with UHC and S-COVID-19-S (p < 0.001); and were significantly lower for those had UHC and interacted with “unchanged/more” physical activity (p < 0.001), or “unchanged/more” drinking (p < 0.001 for only anxiety), or “unchanged/healthier” eating (p < 0.001), and high PB score (p < 0.001), as compared to those without UHC and without S-COVID-19-S, “never/stopped/less” physical activity, drinking, “less healthy” eating, and low PB score, respectively.Conclusion: S-COVID-19-S worsen psychological health in patients with UHC. Physical activity, drinking, healthier eating, and high PB score were protective factors

A Multicentre Molecular Analysis of Hepatitis B and Blood-Borne Virus Coinfections in Viet Nam

Hepatitis B (HBV) infection is endemic in Viet Nam, with up to 8.4 million individuals estimated to be chronically infected. We describe results of a large, multicentre seroepidemiological and molecular study of the prevalence of HBV infection and blood-borne viral coinfections in Viet Nam. Individuals with varying risk factors for infection (n = 8654) were recruited from five centres; Ha Noi, Hai Phong, Da Nang, Khanh Hoa and Can Tho. A mean prevalence rate of 10.7% was observed and levels of HBsAg were significantly higher in injecting drug users (IDUs) (17.4%, n = 174/1000) and dialysis patients (14.3%, n = 82/575) than in lower-risk groups (9.4%; p<0.001). Coinfection with HIV was seen in 28% of HBV-infected IDUs (n = 49/174) and 15.2% of commercial sex workers (CSWs; n = 15/99). HCV infection was present in 89.8% of the HBV-HIV coinfected IDUs (n = 44/49) and 40% of HBV-HIV coinfected CSWs (n = 16/40). Anti-HDV was detected in 10.7% (n = 34/318) of HBsAg positive individuals. Phylogenetic analysis of HBV S gene (n = 187) showed a predominance of genotype B4 (82.6%); genotypes C1 (14.6%), B2 (2.7%) and C5 (0.5%) were also identified. The precore mutation G1896A was identified in 35% of all specimens, and was more frequently observed in genotype B (41%) than genotype C (3%; p<0.0001). In the immunodominant ‘a’ region of the surface gene, point mutations were identified in 31% (n = 58/187) of sequences, and 2.2% (n = 4/187) and 5.3% (n = 10/187) specimens contained the major vaccine escape mutations G145A/R and P120L/Q/S/T, respectively. 368 HBsAg positive individuals were genotyped for the IL28B SNP rs12979860 and no significant association between the IL28B SNP and clearance of HBsAg, HBV viral load or HBeAg was observed. This study confirms the high prevalence of HBV infection in Viet Nam and also highlights the significant levels of blood-borne virus coinfections, which have important implications for hepatitis-related morbidity and development of effective management strategies

Identifying Barriers and Facilitators to Improve Hepatitis C Virus Screening

Background: Birth cohort hepatitis C Virus (HCV) screening recommendations were issued by the Centers for Disease Control and Prevention (CDC) and United States Preventive Services Task Force (USPSTF) in 2012 and 2013, respectively. Despite this, studies have reported low HCV screening rates. Currently, the prevalence of HCV screening rates nationally is unknown and there are no in-depth studies that have evaluated the barriers or facilitators of HCV screening. Our study aimed to fill this gap by 1) conducting a systematic literature review to assess the national HCV screening prevalence, 2) evaluating temporal trends in HCV screening using a large academic center, and 3) identifying barriers and facilitators associated with HCV screening among average-risk baby boomers using geographical information system (GIS) analyses to assess spatial autocorrelation hot spots and cold spots for HCV screening by individual and population-based characteristics. Methods: To accomplish our aims, 1) a systematic literature review was conducted using controlled vocabulary to ascertain the HCV screening prevalence in the U.S. by limiting our search to articles published between January 1, 2012 and March 14, 2019 to coincide with 2012 and 2013 CDC and USPSTF guidelines, respectively for one-time universal testing for HCV among baby boomers (adults born 1945–1965), 2) temporal analyses were conducted covering August 1, 2015 to July 31, 2019 time period for average-risk baby boomers using Cochrane-Armitage test for trend analysis and Joinpoint analysis, and 3) geospatial analyses using geographic information systems’ software were conducted to a) create choropleth maps for HCV antibody test orders and HCV antibody test completion percentages by zip code, b) to measure spatial autocorrelation and to conduct hot spot and cold spot analyses using Global Moran’s Index (GMI) and Getis-Ord GI*, respectively to assess differences in HCV antibody test ordering and HCV antibody test completion percentages using data from the University of South Florida (USF) Healthcare System covering the August 1, 2015 to July 31, 2019 time period among average-risk baby boomers, and c) to evaluate socio-demographic differences between individual and population-based characteristics within hot spot areas (i.e., zip codes with the highest HCV antibody test order and HCV antibody test completion percentages). Patient level data were derived from the USF Healthcare System for the August 1, 2015 to July 31, 2019 time period while population level data were from the American Community Survey (ACS) from the U.S. Census Bureau covering the 2013 to 2017 time period. Results: For Aim 1, after de-duplication, 1,900 articles were identified using four databases (i.e., MEDLINE via PubMed, EMBASE, Web of Science, and the Cochrane Library) for the systematic literature review. After applying inclusion and exclusion criteria, eight articles remained, of which, five were experimental studies (i.e., two randomized clinical trials and three non-randomized trials) and three were observational studies. The risk of bias assessment and the meta-analysis evaluated RCTs and non-randomized trials (i.e., we did not include post-intervention HCV screening data) and observational studies together. We found marked variability in HCV screening estimates among average-risk based baby boomers with a range of 1.4% (95% CI: 1.1%–1.7%) to 12.8% (95% CI: 12.1%–13.5%). The risk of bias assessment revealed high variability in risk of bias. For Aim 2, temporal trends using electronic health records from a large academic center revealed that out of 120,645 opportunities to receive a HCV antibody screening order during 2015–2019, 6,333 HCV antibody orders were placed and of those orders, only 4,259 HCV antibody tests were completed by average-risk baby boomers. The trend by month for HCV ordering percentages of HCV antibody tests during this period of time and showed that the lowest HCV antibody order rate of 1.0% (95% confidence interval [CI]: 0.5%–1.4%) occurred in August 2015. In contrast, the highest HCV antibody order rate of 12.7% (95% CI: 11.4%–14.0%) occurred in May 2017. And by the end of the study period, the HCV antibody order rate was 4.2%. At the start of the period, the HCV screening percentage (HCV antibody test completion) was 0.6% (95% CI: 0.0%–0.9%) in August 2015, at its peak in May 2017, the HCV screening percentage reached 8.8% (95% CI: 7.6%–9.9%), and by the end of the study period, the HCV screening percentage declined to 2.1% (95% CI: 3.1%–2.6%) by July 2019. For Aim 3, spatial autocorrelation showed statistically significant clustering of HCV antibody test orders (GMI = 0.541096; p \u3c 0.000001) and HCV antibody test completion (GMI = 0.445374; p \u3c 0.000001). There were 26 hot spots for HCV antibody test orders among average-risk baby boomers and 22 hot spots were identified for average-risk baby boomers who completed an HCV antibody test. We found that HCV antibody order rates were variable (15.0% to 37.2%) among average-risk baby boomers by zip code within hot spot areas and similarly the HCV screening completion rates also differed ranging between 9.7% to 29.1% by zip code when evaluating hot spots. In general, it was observed that average-risk baby boomers who had the highest percentage (hot spots) of HCV antibody test ordered and the highest percentage (hot spots) of HCV antibody test completed tended to live closer to the healthcare system. Patient level characteristics associated with areas with the highest percentage (hot spots) of HCV antibody test ordered included being female (two-thirds were female), having Medicare insurance (20.1% to 50.0%), having greater than 10+ healthcare visits (15.1% to 41.0%). Similar results were found for hot spot areas with the highest percentage of HCV antibody test. Population-based characteristics associated with the highest (hot spots) HCV antibody test ordered among those who had low poverty levels (4.0% to 32.8%), had low unemployment rates for those age 16+ (1.8% to 12.0%), and had low percentages of people who used public transportation (0.0% to 2.8%), variable percentages of race/ethnicity, and were equally divided by sex. Similar results were found in hot spots for HCV antibody test completion. Conclusions: We found low HCV screening estimates (\u3c 13%) among studies assessed in our systematic literature review in the U.S. Consequently, intervention efforts (e.g. screening and treatment) targeting baby boomers will be important to improve HCV-related disease outcomes. Additionally, to reduce hepatocellular carcinoma related to chronic HCV infections in the U.S., policies and funding are needed to support HCV screening programs to increase uptake of HCV screening. Targeted HCV screening may be warranted for baby boomer populations who are less likely to have an HCV ordered and completed based on patient and population characteristics by using zip code level data by state. Patient level facilitators to HCV antibody test order and completion include being female, having healthcare insurance (Medicare) for baby boomers, having 10+ healthcare visits while population-based characteristics included low poverty percentages, low unemployment, and low use of public transportation, which may be an indicator of economic stability. Future HCV screening interventions should identify facilitators and barriers to HCV screening using a geospatial lens to address HCV screening disparities in vulnerable populations