Grape-associated fungal community patterns persist from berry to wine on a fine geographical scale.

Wine grape fungal community composition is influenced by abiotic factors including geography and vintage. Compositional differences may correlate with different wine metabolite composition and sensory profiles, suggesting a microbial role in the shaping of a wine's terroir, or regional character. While grape and wine-associated fungal community composition has been studied extensively at a regional and sub-regional scale, it has not been explored in detail on fine geographical scales over multiple harvests. Over two years, we examined the fungal communities on Vitis Vinifera cv. Pinot noir grape berry surfaces, in crushed grapes, and in lab spontaneous fermentations from three vineyards within a < 1 km radius in Canada's Okanagan Valley wine region. We also evaluated the effect of winery environment exposure on fungal community composition by sampling grapes crushed and fermented in the winery at commercial scale. Spatiotemporal community structure was evident among grape berry surface, crushed grape and fermentation samples, with each vineyard exhibiting a distinct fungal community signature. Crushed grape fungal populations were richer in fermentative yeast species compared to grape berry surface fungal populations. Our study suggests that, as on a regional level, fungal populations may contribute to fine-scale -terroir,' with significant implications for single-vineyard wines.

Supervised smoking facility access, harm reduction practices, and substance use changes during the COVID-19 pandemic: a community-engaged cross-sectional study.

BACKGROUND: The potential public health benefits of supervised smoking facilities (SSFs) are considerable, and yet implementation of SSFs in North America has been slow. We conducted this study to respond to significant knowledge gaps surrounding SSF utilization and to characterize substance use, harm reduction practices, and service utilization following the onset of the COVID-19 pandemic. METHODS: A questionnaire was self-administered at a single site by 175 clients using an outdoor SSF in Vancouver, Canada, between October-December 2020. Questionnaire responses were summarized using descriptive statistics. Multinomial logistic regression techniques were used to examine factors associated with increased SSF utilization. RESULTS: Almost all respondents reported daily substance use (93% daily use of opioids; 74% stimulants). Most used opioids (85%) and/or methamphetamine (66%) on the day of their visit to the SSF. Respondents reported drug use practice changes at the onset of COVID-19 to reduce harm, including using supervised consumption sites, not sharing equipment, accessing medically prescribed alternatives, cleaning supplies and surfaces, and stocking up on harm reduction supplies. Importantly, 45% of SSF clients reported using the SSF more often since the start of COVID-19 with 65.2% reporting daily use of the site. Increased substance use was associated with increased use of the SSF, after controlling for covariates. CONCLUSIONS: Clients of the SSF reported increasing not only their substance use, but also their SSF utilization and harm reduction practices following the onset of COVID-19. Increased scope and scale of SSF services to meet these needs are necessary.

Whole genome sequencing of Canadian Saccharomyces cerevisiae strains isolated from spontaneous wine fermentations reveals a new Pacific West Coast Wine clade.

Vineyards in wine regions around the world are reservoirs of yeast with oenological potential. Saccharomyces cerevisiae ferments grape sugars to ethanol and generates flavor and aroma compounds in wine. Wineries place a high-value on identifying yeast native to their region to develop a region-specific wine program. Commercial wine strains are genetically very similar due to a population bottleneck and in-breeding compared to the diversity of S. cerevisiae from the wild and other industrial processes. We have isolated and microsatellite-typed hundreds of S. cerevisiae strains from spontaneous fermentations of grapes from the Okanagan Valley wine region in British Columbia, Canada. We chose 75 S. cerevisiae strains, based on our microsatellite clustering data, for whole genome sequencing using Illumina paired-end reads. Phylogenetic analysis shows that British Columbian S. cerevisiae strains cluster into 4 clades: Wine/European, Transpacific Oak, Beer 1/Mixed Origin, and a new clade that we have designated as Pacific West Coast Wine. The Pacific West Coast Wine clade has high nucleotide diversity and shares genomic characteristics with wild North American oak strains but also has gene flow from Wine/European and Ecuadorian clades. We analyzed gene copy number variations to find evidence of domestication and found that strains in the Wine/European and Pacific West Coast Wine clades have gene copy number variation reflective of adaptations to the wine-making environment. The "wine circle/Region B", a cluster of 5 genes acquired by horizontal gene transfer into the genome of commercial wine strains is also present in the majority of the British Columbian strains in the Wine/European clade but in a minority of the Pacific West Coast Wine clade strains. Previous studies have shown that S. cerevisiae strains isolated from Mediterranean Oak trees may be the living ancestors of European wine yeast strains. This study is the first to isolate S. cerevisiae strains with genetic similarity to nonvineyard North American Oak strains from spontaneous wine fermentations.

Peer Support Specialists: An Underutilized Resource in the Criminal Justice System for Opioid Use Disorder Management?

In the wake of North America's worsening overdose crisis, the overrepresentation of individuals incarcerated with an opioid use disorder (OUD) constitutes a population at an incredibly high risk for adverse health outcomes, including death. In response, a number of important initiatives such as the provision of opioid agonist therapy to individuals with opioid addiction while incarcerated have been implemented. Although improving access to evidence-based treatment for OUD is an obvious urgent need, equally important is the need to implement novel interventions to help reduce morbidity and mortality among this high-risk group. Peer support specialists (ie, individuals with lived or shared experience) have previously been demonstrated to effectively help clients navigate the healthcare system, reintegrate within their community, and successfully adhere to their individual treatment and recovery goals. Given the known association between individuals with an OUD and exposure to the criminal justice system, routine inclusion of peer support specialists as part of the addiction interdisciplinary care team in these settings may be an effective opportunity to improve health outcomes and prevent death among incarcerated individuals with an OUD.

HIV and Incarceration: Implications for HIV-Positive People Who Use Illicit Drugs During a Seek, Test, Treat, and Retain Initiative in Canada.

People who use illicit drugs (PWUD) face increased exposure to the criminal justice system and disproportionate burdens of HIV infection. This article investigated the effects of incarceration on HIV cascade of care-related measures in a setting with a community-wide seek, test, treat, and retain (STTR) initiative. Using a multivariable logistic regression analysis of 935 HIV-positive PWUD between 2005 and 2017, this article showed a negative relationship between periods of incarceration and two measures of engagement in clinical care for HIV among PWUD: recent dispensation of antiretroviral therapy and suppression of HIV viral load. These findings suggest the benefits of STTR-based efforts are limited by exposure to the criminal justice system and highlight the need for additional supports for PWUD with HIV exposed to the criminal justice system.

Characterization of Sub-Regional Variation in Saccharomyces Populations and Grape Phenolic Composition in Pinot Noir Vineyards of a Canadian Wine Region.

Wine is a product of grape juice fermentation by yeast. Terroir is a term that encompasses all environmental factors and interactions at a specific geographical site, resulting in the development of regional-specific microbial strains and grape metabolites. In this study we determine the distribution of vineyard-associated wine yeast strains and characterize the flavonoid profile of Pinot Noir grapes among 3 sub-regions in the Okanagan Valley (OV), a major wine region in British Columbia, Canada. Pinot Noir grape samples were collected from 13 vineyards among 3 sub-regions of the OV, namely Kelowna (KE), Naramata-Penticton (NP) and Oliver-Osoyoos (OO), within a week prior to the winery harvesting date in 2016 and 2017. A total of 156 spontaneous Pinot Noir fermentations were conducted and vineyard-associated Saccharomyces strains were isolated from fermentations that reached two-thirds sugar depletion. Using microsatellite genotyping, we identified 103 Saccharomyces cerevisiae strains and 9 Saccharomyces uvarum strains. We also identified Saccharomyces paradoxus in one vineyard using ITS sequencing. We developed a microsatellite database of 160 commercial S. cerevisiae strains to determine the identity of the isolated strains and we include the database herein. Commercial strains were widely distributed across the three sub-regions. Forty-two of our 103 S. cerevisiae strains were equivalent or highly similar to commercial strains whereas the remaining 61 were considered as 'unknown' strains. Two S. uvarum strains were previously isolated in other OV studies and none matched the S. uvarum commercial strain BMV58. S. cerevisiae population structure was driven by sub-region, although S. cerevisiae populations did not differ significantly across vintages. S. uvarum and S. paradoxus were only identified in the 2017 vintage, demonstrating dynamic wine yeast populations between vintages. We found that the flavonoid profile of Pinot Noir grapes from the same 13 vineyards was also affected by sub-regional terroir. The anthocyanin content was lower and the proportion of methoxylated anthocyanins and flavonols was higher in Pinot Noir grapes from OO, the warmer sub-region as compared to KE, the cooler sub-region. Our study demonstrates that both yeast populations and metabolites associated with the Pinot Noir variety have sub-regional variation within a viticultural area.

N-Terminal Pro-B-Type Natriuretic Peptide (NT-proBNP) Immunoreactivity Is Reduced After 6 Years of Storage at -70 °C.

BACKGROUND: Clinical trial samples may be stored frozen for prolonged periods before analysis, which can reduce the immunoreactivity of numerous analytes, particularly peptides. We sought to determine the effect of 6 years of frozen storage on serum N-terminal pro-B-type natriuretic peptide (NT-proBNP). METHODS: NT-proBNP was measured from serum samples taken from 99 different patients enrolled in the CanPREDDICT study after <1 year of storage at -70 °C using the Roche first-generation NT-proBNP assay on an e411 instrument. Separate aliquots of the same samples were analyzed after an additional 6 years of storage at -70 °C using the Roche second-generation assay on an e601 instrument. RESULTS: The median NT-proBNP immunoreactivity for the first measurement was 572 pg/mL (interquartile range [IQR] 205-1606, range 49-12820), while after an additional 6 years of storage at -70 °C, this value decreased to 526 pg/mL (IQR 181-1338, range 18-12880), resulting in a median percent difference of -7% (IQR -10.6% to -3.4%, P < 0.001). CONCLUSIONS: We report findings consistent with trends seen in previous work but have investigated the effect of a much longer storage period. Larger percent decreases in NT-proBNP reaching statistical significance are seen, although the median difference is still <10%.