Resurrection of 2'-5'-oligoadenylate synthetase 1 (OAS1) from the ancestor of modern horseshoe bats blocks SARS-CoV-2 replication.

The prenylated form of the human 2'-5'-oligoadenylate synthetase 1 (OAS1) protein has been shown to potently inhibit the replication of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), the virus responsible for the Coronavirus Disease 2019 (COVID-19) pandemic. However, the OAS1 orthologue in the horseshoe bats (superfamily Rhinolophoidea), the reservoir host of SARS-related coronaviruses (SARSr-CoVs), has lost the prenylation signal required for this antiviral activity. Herein, we used an ancestral state reconstruction approach to predict and reconstitute in vitro, the most likely OAS1 protein sequence expressed by the Rhinolophoidea common ancestor prior to its prenylation loss (RhinoCA OAS1). We exogenously expressed the ancient bat protein in vitro to show that, unlike its non-prenylated horseshoe bat descendants, RhinoCA OAS1 successfully blocks SARS-CoV-2 replication. Using protein structure predictions in combination with evolutionary hypothesis testing methods, we highlight sites under unique diversifying selection specific to OAS1's evolution in the Rhinolophoidea. These sites are located near the RNA-binding region and the C-terminal end of the protein where the prenylation signal would have been. Our results confirm that OAS1 prenylation loss at the base of the Rhinolophoidea clade ablated the ability of OAS1 to restrict SARSr-CoV replication and that subsequent evolution of the gene in these bats likely favoured an alternative function. These findings can advance our understanding of the tightly linked association between SARSr-CoVs and horseshoe bats.

A microfluidic system that replicates pharmacokinetic (PK) profiles in vitro improves prediction of in vivo efficacy in preclinical models.

Test compounds used on in vitro model systems are conventionally delivered to cell culture wells as fixed concentration bolus doses; however, this poorly replicates the pharmacokinetic (PK) concentration changes seen in vivo and reduces the predictive value of the data. Herein, proof-of-concept experiments were performed using a novel microfluidic device, the Microformulator, which allows in vivo like PK profiles to be applied to cells cultured in microtiter plates and facilitates the investigation of the impact of PK on biological responses. We demonstrate the utility of the device in its ability to reproduce in vivo PK profiles of different oncology compounds over multiweek experiments, both as monotherapy and drug combinations, comparing the effects on tumour cell efficacy in vitro with efficacy seen in in vivo xenograft models. In the first example, an ERK1/2 inhibitor was tested using fixed bolus dosing and Microformulator-replicated PK profiles, in 2 cell lines with different in vivo sensitivities. The Microformulator-replicated PK profiles were able to discriminate between cell line sensitivities, unlike the conventional fixed bolus dosing. In a second study, murine in vivo PK profiles of multiple Poly(ADP-Ribose) Polymerase 1/2 (PARP) and DNA-dependent protein kinase (DNA-PK) inhibitor combinations were replicated in a FaDu cell line resulting in a reduction in cell growth in vitro with similar rank ordering to the in vivo xenograft model. Additional PK/efficacy insight into theoretical changes to drug exposure profiles was gained by using the Microformulator to expose FaDu cells to the DNA-PK inhibitor for different target coverage levels and periods of time. We demonstrate that the Microformulator enables incorporating PK exposures into cellular assays to improve in vitro-in vivo translation understanding for early therapeutic insight.

A multicenter evaluation of a novel microfluidic rapid AST assay for Gram-negative bloodstream infections.

Common phenotypic methods for antimicrobial susceptibility testing (AST) of bacteria are slow, labor intensive, and display considerable technical variability. The QuickMIC system provides rapid AST using a microfluidic linear gradient. Here, we evaluate the performance of QuickMIC at four different laboratories with regard to speed, precision, accuracy, and reproducibility in comparison to broth microdilution (BMD). Spiked (n = 411) and clinical blood cultures (n = 148) were tested with the QuickMIC Gram-negative panel and compared with BMD for the 12 on-panel antibiotics, and 10 defined strains were run at each site to measure reproducibility. Logistic and linear regression analysis was applied to explore factors affecting assay performance. The overall essential agreement and categorical agreement between QuickMIC and BMD were 95.6% and 96.0%, respectively. Very major error, major error, and minor error rates were 1.0%, 0.6%, and 2.4%, respectively. Inter-laboratory reproducibility between the sites was high at 98.9% using the acceptable standard of ±1 twofold dilution. The mean in-instrument analysis time overall was 3 h 13 min (SD: 29 min). Regression analysis indicated that QuickMIC is robust with regard to initial inoculum and delay time after blood culture positivity. We conclude that QuickMIC can be used to rapidly measure MIC directly from blood cultures in clinical settings with high reproducibility, precision, and accuracy. The microfluidics-generated linear gradient ensures high reproducibility between laboratories, thus allowing a high level of trust in MIC values from single testing, at the cost of reduced measurement range compared to BMD. IMPORTANCE: Increasing antimicrobial resistance underscores the need for new diagnostic solutions to guide therapy, but traditional antimicrobial susceptibility testing (AST) is often inadequate in time-critical diseases such as sepsis. This work presents a novel and rapid AST system with a rapid turnaround of results, which may help reduce the time to guided therapy, possibly allowing early de-escalation of broad-spectrum empirical therapy as well as rapid adjustments to treatments when coverage is lacking.

If They'd Said You Should Only Drink Five Units I'd Have Listened: A Mixed Methods Study of Alcohol Consumption Following a Diagnosis of Breast Cancer.

OBJECTIVES: As part of a wider study describing the impact of a breast cancer diagnosis on lifestyle behaviours, this paper describes the impact of a breast cancer diagnosis on alcohol consumption and factors influencing consumption. METHODS: Cross-sectional online survey of 140 people (138 women) and interviews with 21 women diagnosed with breast cancer in the last 10 years. RESULTS: Of the 100 survey participants who drank alcohol 25% were drinking at increasing or higher risk levels and 17% strongly wanted to change their drinking behaviour. The habitual aspects of alcohol consumption were the strongest predictor of current alcohol consumption behaviours. Social norms and perceptions about conflicting information were substantial barriers to change. CONCLUSIONS: Breast cancer survivors need accurate information about the risks of alcohol consumption and guidelines in order to make informed decisions about making changes to their behaviour. Interventions to support breast cancer survivors to reduce alcohol consumption need to focus on the development of healthy habits and may benefit from a focus which includes partners and friends.

Discrimination, gender dysphoria, drinking to cope, and alcohol harms in the UK trans and non-binary community.

Transgender (trans) and non-binary people may be at increased risk of alcohol harms, but little is known about motives for drinking in this community. This study explored the relationship between risk of alcohol dependence, experience of alcohol harms, drinking motives, dysphoria, and discrimination within a United Kingdom sample of trans and non-binary people with a lifetime history of alcohol use. A cross-sectional survey was co-produced with community stakeholders and administered to a purposive sample of trans and non-binary people from 1 February until 31 March 2022. A total of 462 respondents were included-159 identified as non-binary and/or genderqueer (identities outside the man/woman binary), 135 solely as women, 63 solely as men, 15 as another gender identity, 90 selected multiple identities. Higher levels of reported discrimination were associated with higher risk of dependence and more reported harms from drinking. Coping motives, enhancement motives, and drinking to manage dysphoria were associated with higher Alcohol Use Disorders Identification Test scores. Social, coping, and enhancement motives alongside discrimination and drinking to have sex were associated with harms. The relationship between discrimination and risk of dependence was mediated by coping motives and drinking to manage dysphoria. Further to these associations, we suggest that reducing discrimination against trans and non-binary communities might reduce alcohol harms in this population. Interventions should target enhancement motives, coping motives and gender dysphoria. Social and enhancement functions of alcohol could be replaced by alcohol free supportive social spaces.

Drinking and mental health in middle adulthood: exploring the impact of wellbeing, mental health literacy, and drinking motives on risk of alcohol dependence.

OBJECTIVES: Risky drinking is a concern among UK-based middle-aged adults. We aimed to explore the relationship between risky drinking, drinking motives, wellbeing, and mental health literacy (MHL). METHOD: Cross-sectional analysis of online survey data completed by 193 UK-based adults aged 40-65 who drank alcohol, incorporating the Alcohol Use Disorders Identification Test (AUDIT); Drinking Motives Questionnaire-Revised (DMQ-R); Warwick-Edinburgh Mental Wellbeing Scale (WEMWBS); Mental Health Literacy Scale (MHLS) and demographic questions. RESULTS: Coping, enhancement and conformity motives and gender significantly predicted higher AUDIT scores (measuring risky drinking). Enhancement motives were found to mediate the relationship between the self-help component of MHL and AUDIT scores, while coping motives mediated the association between wellbeing and AUDIT scores. CONCLUSION: Findings support research emphasising the influence of drinking motives on risky drinking and highlights how low wellbeing may interact with coping motives to explain risky drinking among middle-aged adults, particularly men. Interventions supporting individuals to understand the relationship between drinking motives and risky drinking, develop adaptive coping strategies, and address the causes of low wellbeing, may be beneficial. However, as the sample was 84% ethnically White, 64% women, 85% educated to at least undergraduate level, and reported a relatively high mean socioeconomic status (6.98 out of 10), the results may not generalise beyond these groups. Future research should use stratified sampling to increase generalisability, as well as exploring whether alcohol-specific, component-specific, or disorder-specific MHL is associated with risky drinking and wellbeing.

Regrets, I've Had a Few: Exploring Factors Associated with Getting Drunk and Regret in an International Study of People Who Drink Alcohol.

BACKGROUND: Excessive alcohol consumption is often followed by feelings of regret. This study aimed to explore country differences in experiences of drunkenness and regrets and predictors of experiencing a greater number of regrettable drinking occasions. METHODS: This study draws on a sample of 82,821 respondents from 31 countries who completed the 2020 Global Drug Survey. Respondents were asked to report how many times in the last year they had been drunk, how many of those times they felt regret afterwards and to complete a range of sociodemographic measures. RESULTS: In the last 12 months, the median times drunk was 6 and the median number of regretted occasions was 2. There was an inverse relationship between times drunk and regret. Respondents who got drunk more often regretted it a smaller percentage of the time than those who got drunk less often. Respondents from Argentina and Colombia regretted being drunk the most and Denmark the least. Being younger, in higher AUDIT categories were associated with more times drunk. Being a woman, having mental health conditions were associated with more regretted occasions.Discussion and conclusions: Country variations may reflect relative acceptability of being drunk. Those who drink more, per occasion, may become accustomed to the consequences and feel fewer regrets. Interventions promoting reduced alcohol consumption may benefit from encouraging people to consider their future regret following a drinking occasion but should account for lower levels of regret in those who get drunk more often.

Nosocomial outbreak in a respiratory ward caused by the SARS-CoV-2 Omicron BA 5.2.1 subvariant associated with non-severe illness in vaccinated patients.

In this short report, we describe an outbreak of COVID-19 caused by Omicron subvariant BA.5.2.1 in highly vaccinated patients in a respiratory ward in a large acute general hospital in North West London, United Kingdom. The attack rate was high (14/33 (42%)) but the clinical impact was relatively non-severe including in patients who were at high risk of severe COVID-19. Twelve of fourteen patients had COVID-19 vaccinations. There was only one death due to COVID-19 pneumonitis. The findings of this outbreak investigation suggest that while the transmissibility of Omicron BA.5.2.1 subvariant is high, infections caused by this strain are non-severe in vaccinated patients, even if they are at high risk of severe COVID-19 infection.

Characterization of microtubule-associated protein tau isoforms and Alzheimer's disease-like pathology in normal sheep (Ovis aries): relevance to their potential as a model of Alzheimer's disease.

Alzheimer's disease is a chronic neurodegenerative disease that accounts for up to 80% of all dementias. Characterised by deteriorations of memory and cognitive function, the key neuropathological features are accumulations of ß-amyloid and hyperphosphorylated tau, as 'plaques' and 'tangles', respectively. Despite extensive study, however, the exact mechanism underlying aggregate formation in Alzheimer's disease remains elusive, as does the contribution of these aggregates to disease progression. Importantly, a recent evaluation of current Alzheimer's disease animal models suggested that rodent models are not able to fully recapitulate the pathological intricacies of the disease as it occurs in humans. Therefore, increasing attention is being paid to species that might make good alternatives to rodents for studying the molecular pathology of Alzheimer's disease. The sheep (Ovis aries) is one such species, although to date, there have been few molecular studies relating to Alzheimer's disease in sheep. Here, we investigated the Alzheimer's disease relevant histopathological characteristics of 22 sheep, using anti-ß-amyloid (Abcam 12267 and mOC64) and phosphorylation specific anti-tau (AT8 and S396) antibodies. We identified numerous intraneuronal aggregates of both ß-amyloid and tau that are consistent with early Alzheimer's disease-like pathology. We confirmed the expression of two 3-repeat (1N3R, 2N3R) and two 4-repeat (1N4R, 2N4R) tau isoforms in the ovine brain, which result from the alternative splicing of two tau exons. Finally, we investigated the phosphorylation status of the serine396 residue in 30 sheep, and report that the phosphorylation of this residue begins in sheep aged as young as 2 years. Together, these data show that sheep exhibit naturally occurring ß-amyloid and tau pathologies, that reflect those that occur in the early stages of Alzheimer's disease. This is an important step towards the validation of the sheep as a feasible large animal species in which to model Alzheimer's disease.

Experience and perpetration of intimate partner violence and abuse by gender of respondent and their current partner before and during COVID-19 restrictions in 2020: a cross-sectional study in 13 countries.

BACKGROUND: Intimate partner violence and abuse (IPVA) includes controlling behaviours, psychological, physical, sexual and financial abuse. Globally, surveys and emergency services have recorded an increase in IPVA since restrictions were imposed to limit COVID-19 transmission. Most studies have only included heterosexual women. METHODS: Data from the Global Drug Survey (an annual, anonymous, online survey collecting data on drug use) Special Edition were analysed to explore the impact of COVID-19 on people's lives, including their intimate relationships. Five relationship groupings were created using respondents' lived gender identity: women partnered with men (46.9%), women partnered with women (2.1%), men partnered with men (2.9%), men partnered with women (47.2%), and partnerships where one or both partners were non-binary (1%). Self-reported experience and perpetration of IPVA in the past 30 days before (February) and during COVID-19 restrictions (May or June) in 2020 (N = 35,854) was described and compared for different relationship groupings using Fishers Exact Tests. Changes in IPVA during restrictions were assessed using multivariable logistic regression. RESULTS: During restrictions, 17.8 and 16.6% of respondents had experienced or perpetrated IPVA respectively; 38.2% of survivors and 37.6% of perpetrators reported this had increased during restrictions. Greater proportions of non-binary respondents or respondents with a non-binary partner reported experiencing or perpetrating IPVA (p < .001) than other relationship groupings. 22.0% of respondents who were non-binary or had a non-binary partner, 19.5% of men partnered with men, 18.9% of men partnered with women, 17.1% of women partnered with women and 16.6% of women partnered with men reported experiencing IPVA. Respondents with higher psychological distress, poor coping with pandemic-related changes, relationship tension and changes (increases or increases and decreases) in alcohol consumption reported increased experience of IPVA during restrictions. CONCLUSIONS: This study confirmed that IPVA can occur in all intimate relationships, regardless of gender of the perpetrator or survivor. Non-binary respondents or respondents with non-binary partners reported the highest use and experience of IPVA. Most IPVA victim support services have been designed for heterosexual, cisgender women. IPVA support services and perpetrator programmes must be tailored to support all perpetrators and survivors during the pandemic and beyond, regardless of their sexual or gender identity.

Shouldn't We Know This Already? UK Women's Views About Communicating the Link Between Alcohol Consumption and Risk of Breast Cancer.

Alcohol is a causal factor in about 10% of breast cancer (BCa) cases, but awareness of this link is low. This study explored how to raise awareness and inform the development of an intervention using the COM-B model (capability, opportunity, motivation, behavior) framework. Eight online focus groups were conducted with 36 participants (6 expert stakeholders,and 30 women aged 40-65). Participants reflected on a package of information about alcohol and BCa and discussed how to impart this information and encourage women to reduce drinking. Thematic analysis of focus group transcripts was undertaken. Three themes were identified: understanding ineffective messaging; transitions and challenges; and message acceptability. Current health information about alcohol was perceived as judgmental and BCa was put down to chance. Mid-life consisted of many challenges that could lead to increased consumption, but menopause transition may be a key moment for alcohol reduction. Barriers and enablers to communicating risk information and encouraging alcohol reduction were mapped onto the COM-B model. Psychological capability (relating to knowledge), social opportunity (in the form of social pressure) and automatic motivation (relating to drinking to cope) were barriers to behavior change.  These will be targeted in an alcohol reduction intervention. It is important to tailor information to women's experiences, taking into account the social benefits of drinking, and encourage the development of healthy coping strategies. Acceptable intervention messages may include personal stories, clear statistics, and suggest healthy alternatives to drinking. It is vital that messaging does not appear judgmental or patronizing.

Frailty in Very Elderly Patients is Not Associated with Adverse Surgical or Oncological Outcomes in Extremity Surgery for Soft Tissue Sarcoma.

BACKGROUND: While surgery remains the mainstay of treatment for limb sarcoma, extreme old age is a relative contraindication to oncological surgery. METHODS: Patients >80 years referred with primary extremity soft-tissue sarcoma (ESTS) between 2007 and 2016 were retrospectively reviewed. Prognostic variables, including ASA status and Clinical Frailty Scores, were collected. Endpoints were perioperative morbidity, locoregional (LRR) and distant recurrence (DR), disease-specific survival (DSS) adjusted using competing risk modelling, and overall survival (OS). RESULTS: A total of 141 primary tumours were identified, with 116 undergoing resections. Main motives for nonoperative management were severe frailty or significant comorbidity (56.0%). The operative group had a median age of 84 (range 80-96) years and median follow-up of 16 months (range 0-95). 45.7% of patients received radiotherapy. Median hospital stay was 7 (range 0-40) days, with frailty (p = 0.25) and ASA (p = 0.28) not associated with prolonged admission. 12.9% developed significant complications, with one perioperative mortality. 24.1% had LRR, occurring at a median of 14.5 months. All patients with reported DR (28.4%), except one, died of their disease. Frailty did not confer a significant difference in adjusted LRFS (p = 0.95) and DMFS (p = 0.84). One- and 5-year adjusted DSS and OS was 87.0% versus 74.9% and 62.3% versus 27.4%, respectively. Frailty (CFS ≥4) was associated with worse OS (hazard ratio [HR] 2.49; 95% confidence interval [CI] 1.51-4.12; p < 0.001), however not with adjusted DSS (p = 0.16). Nonoperative management conferred a 1- and 5-year adjusted DSS was 58.3% and 44.4%, respectively. CONCLUSIONS: Extremity surgery for sarcoma is well tolerated in the frail very elderly population with low morbidity and comparable oncological outcomes.

Testing socioeconomic status and family socialization hypotheses of alcohol use in young people: A causal mediation analysis.

INTRODUCTION: The effect of socioeconomic status on adolescent substance abuse may be mediated by family socialization practices. However, traditional mediation analysis using a product or difference method is susceptible to bias when assumptions are not addressed. We aimed to use a potential outcomes framework to assess assumptions of exposure-mediator interaction and of no confounding of the results. METHOD: We revisited a traditional mediation analysis with a multiple mediator causal mediation approach using data from 17,761 Norwegian young people (13-18 years), 51% female. Data were collected through a print questionnaire. Socioeconomic status was operationalized as parental education and employment status (employed or receiving welfare); drinking behavior as the frequency of alcohol consumption and frequency of intoxication in the past year; and socialization practices as general parenting measures, alcohol-related parental permissiveness, and parent drinking behavior. RESULTS: There was no consistent evidence of exposure-mediator interaction. Formal sensitivity analysis of mediator-outcome confounding was not possible in the multiple mediator model, and this analysis supported the hypothesis that socioeconomic status effects on adolescent substance abuse are fully mediated by family socialization practices, with apparently stronger effects in younger age groups observed in plots. CONCLUSION: We found that the effect of socioeconomic status on adolescent substance abuse was fully mediated by family socialization practices. While our analysis provides more rigorous support for causal inferences than past work, we could not completely rule out the possibility of unmeasured confounding.

Connecting Through Dance: Understanding Conscious Clubbing Event Experiences.

Individuals abstaining from alcohol consumption frequently find themselves in contexts encouraging consumption, with limited alternative social interaction opportunities. Conscious clubbing events exclude alcohol and drugs, possibly providing valuable social connections, but little is known about event benefits. Twelve conscious clubbing event attendees and facilitators aged 25-55 from across Europe participated in semi-structured photo-elicitation interviews, which were analysed using thematic analysis. Findings suggested that conscious clubbing appears to enhance health, healing and growth, aiding recovery from substance dependency and trauma. Positioned as a modern ritual, symbolic and ritualistic preparations and experiences promoted storytelling and self-discovery. Connecting through synchronised but unchoreographed movement, participants engaged in powerful journeys and transformative experiences detached from life constraints. Inclusive digital and face-to-face conscious clubbing communities provided a sense of meaning and belonging away from substance use pressures and harms, particularly for those marginalised within society. Findings provide important and in-depth insights, including novel harm prevention implications. Future work should consider participatory barriers, event sustainability and misconceptions to increase event availability and participation.

Taking Action in Student Harassment Situations: Application of the Behaviour Change Wheel to Bystander Intervention.

This study applied the Behaviour Change Wheel (BCW) to understand barriers and facilitators to bystander behaviors in UK students. The BCW includes detailed examination of the capabilities, opportunities, and motivations involved in behaviors. Two surveys (n = 515; n = 201) and a focus group study (n = 12) were undertaken. Capability to intervene may be influenced by confidence and beliefs about physical ability and safety. Students appeared to have the physical opportunity to intervene, but social opportunity might be influenced by cultural norms. Motivations might be influenced by beliefs as well as inherent stereotypes about perpetrators and victims. Behavior change techniques (BCTs) such as instruction on how to perform the behavior, reattribution and creating a valued self-identity should be applied to overcome these barriers. A logic model to theorize the change processes underlying bystander behaviors in this population offers a new perspective on what needs to be addressed in interventions.

Childhood adversities among students at an English University: A latent class analysis.

University students routinely participate in research, including research on trauma and adversity, but the unique implications of trauma and adversity for educational and developmental outcomes for this group have received less attention. This study surveyed first year undergraduate students at an urban university located in the most ethnically diverse district in England, with the second highest poverty rate. Of 7,110 students, 858 responded; a response rate of 12%. The survey included thirteen questions about adverse events and circumstances before age 18. Four in five students (79%) reported at least one adversity, 51% reported three or more, and 20% reported at least six. Female students reported a higher mean number of adversities than men, but men were significantly more likely to report having been "attacked, stabbed, shot or robbed by threat." Where comparisons were possible, rates were higher than for the general population or for the only other UK university survey. A Latent Class Analysis produced four groupings. Besides the 'No adversity' (36%) and "Intermediate" (46%) classes, there were two "High adversity" groups, differentiated by high (12%) or moderate (6%) adversities related to cohabitation (parental separation, lived with depressed person, lived with alcohol/drug user, and lived with incarcerated person). Higher rates of adversities, and latent class membership, were related to predictions that one would not complete one's degree. Implications and next steps are discussed.

The impact of adverse childhood experiences and recent life events on anxiety and quality of life in university students.

Adverse childhood experiences (ACEs) have been repeatedly associated with a wide range of physical and mental health issues. Research has indicated high levels of anxiety and depression among university students, and a few studies have documented the relationship between ACEs and anxiety in the university student population. This study surveyed first year students at a university located in the most ethnically diverse district in England, with the second highest poverty rate. Eight hundred and fifty-eight responded; a response rate of 12%. The survey included questions about adverse childhood events, recent life stressors, current deprivation, quality of life, positive physical health and positive mental health, and used the Generalized Anxiety Disorder Scale, a standardised measure. Thirty-seven percent of the responding students met the diagnosis for generalised anxiety disorder. In a multivariate multiple regression model, life stressors and childhood adversities were significantly associated with higher levels of anxiety and lower levels of physical and mental health. Only childhood adversities significantly predicted lower levels of quality of life. The findings highlight the importance of considering adverse childhood experiences in enhancing the wellbeing of the student population. Given the demographics of the student population at the University of East London, the high rates of ACEs and anxiety are likely to partially reflect poverty and racism. The implications of the findings for trauma-informed policies and practices in universities are discussed. Supplementary Information: The online version contains supplementary material available at 10.1007/s10734-021-00774-9.

Tumor necrosis is significantly associated with reduced recurrence-free survival after curative resection of gastrointestinal stromal tumors.

BACKGROUND OBJECTIVES: The impact of tumor necrosis as a prognostic factor in gastrointestinal stromal tumor (GISTs) is still debated. The objective was to determine whether tumor necrosis is an independent risk factor for survival in patients with GISTs. METHODS: Patients undergoing surgery for primary GIST from March 2003 to October 2018 at two sarcoma referral centers were retrospectively identified. Patients who received neoadjuvant imatinib were excluded. Multivariable Cox regression models were produced, to assess whether tumor necrosis was an independent predictor of either overall or recurrence-free survival. RESULTS: Forty-one out of 195 (21.0%) patients had tumor necrosis. Tumor necrosis was associated with a significantly higher modified National Institute of Health risk score, with 29 out of 41 (70.7%) patients with necrosis classified as high risk, compared to 52 out of 153 (34.0%) without (p < .001). Tumor necrosis was found to be independently predictive of recurrence-free survival (hazard ratio: 5.26, 95% CI: 2.62-10.56, p < .001) on multivariable analysis. At 5 years, 44.3% of patients with necrosis had either died or developed recurrence, compared to 9.9% of those without. CONCLUSION: Tumor necrosis is an independent predictor of recurrence-free survival in patients with operable GISTs. It should be routinely reported by pathologists, and used by clinicians when counseling patients and deciding on adjuvant therapy.

Temporal dynamics of trematode intermediate snail host environmental DNA in small water body habitats.

Environmental DNA (eDNA) surveying has potential to become a powerful tool for sustainable parasite control. As trematode parasites require an intermediate snail host that is often aquatic or amphibious to fulfil their lifecycle, water-based eDNA analyses can be used to screen habitats for the presence of snail hosts and identify trematode infection risk areas. The aim of this study was to identify climatic and environmental factors associated with the detection of Galba truncatula eDNA. Fourteen potential G. truncatula habitats on two farms were surveyed over a 9-month period, with eDNA detected using a filter capture, extraction and PCR protocol with data analysed using a generalized estimation equation. The probability of detecting G. truncatula eDNA increased in habitats where snails were visually detected, as temperature increased, and as water pH decreased (P < 0.05). Rainfall was positively associated with eDNA detection in watercourse habitats on farm A, but negatively associated with eDNA detection in watercourse habitats on farm B (P < 0.001), which may be explained by differences in watercourse gradient. This study is the first to identify factors associated with trematode intermediate snail host eDNA detection. These factors should be considered in standardized protocols to evaluate the results of future eDNA surveys.

Where and What You Drink Is Linked to How Much You Drink: An Exploratory Survey of Alcohol Use in 17 Countries.

BACKGROUND: This paper aimed to explore the differences in subjective experiences of intoxication depending on drinking location and drink type. Methods: Data came from 32,194 respondents to The Global Drug Survey (GDS) 2015, an annual, cross-sectional, online survey. Respondents selected their usual drinking location (home alone: home with partner/family: house parties: pubs/bars or clubs) and usual drink (wine; beer/cider/lager; spirits or alcopops/coolers). They indicated how many drinks they required to reach three stages of intoxication (feeling the effects; an ideal stage of intoxication; and the tipping point) and how frequently they reached each stage. Results: Drink type affected grams of alcohol reported to reach the tipping point: 109 gm wine, 127 gm alcopops, 133 gm of beer, and 134 gm of spirts. Respondents who drank at home alone, or in clubs reached their tipping point more frequently compared to other locations. Conclusions: Where people drink, and the type of alcohol they drink, affected the amount of alcohol reported to reach different stages of intoxication. Understanding why different drinking locations, and drink types lead to a need for greater consumption to reach an ideal state of drunkenness, such as social cues from other people who drink, may enable people to reduce their drinking.