Identification of USP9X as a leukemia susceptibility gene.

We recently reported that children with multiple birth defects have a significantly higher risk of childhood cancer. We performed whole-genome sequencing on a cohort of probands from this study with birth defects and cancer and their parents. Structural variant analysis identified a novel 5 kb de novo heterozygous inframe deletion overlapping the catalytic domain of USP9X in a female proband with multiple birth defects, developmental delay, and B-cell acute lymphoblastic leukemia (B-ALL). Her phenotype was consistent with female-restricted X-linked syndromic intellectual developmental disorder-99 (MRXS99F). Genotype-phenotype analysis including previously reported female probands (n = 42) demonstrated that MRXS99F probands with B-ALL (n = 3) clustered with subjects with loss-of-function (LoF) USP9X variants and multiple anomalies. The cumulative incidence of B-ALL among these female probands (7.1%) was significantly higher than an age- and sex-matched cohort (0.003%) from the Surveillance, Epidemiology, and End Results database (P < .0001, log-rank test). There are no reports of LoF variants in males. Males with hypomorphic missense variants have neurodevelopmental disorders without birth defects or leukemia risk. In contrast, in sporadic B-ALL, somatic LoF USP9X mutations occur in both males and females, and expression levels are comparable in leukemia samples from both sexes (P = .54), with the highest expressors being female patients with extra copies of the X-chromosome. Overall, we describe USP9X as a novel female-specific leukemia predisposition gene associated with multiple congenital, neurodevelopmental anomalies, and B-ALL risk. In contrast, USP9X serves as a tumor suppressor in sporadic pediatric B-ALL in both sexes, with low expression associated with poorer survival in patients with high-risk B-ALL.

Youth perceptions of Brand variant names on standardised cigarette packs, and responses to replacing these with numbers: a focus group study in Britain.

Tobacco companies use brand variant name on cigarette packaging to differentiate, and create interest in, their products. We explored young peoples' reactions to brand variant names on cigarette packs and perceptions of replacing these with numbers, a proposed policy in Turkey. Twelve focus groups, segmented by gender, age (11-12, 13-14, 15-16) and social grade (ABC1, C2DE), were conducted across Britain from May-July 2018 (n = 89). Participants were asked what they thought about brand names in general, and on cigarette packs, and perceptions of replacing the brand variant name on cigarette packs with a number. Brand (variant) name was considered important for products, including cigarettes, and thought to communicate information about the product, image, price, and taste, and encourage purchase. Although replacing brand variant names on cigarette packs with numbers caused confusion, several participants mentioned that it would eliminate any remaining marketing power that the pack may have. They thought that numbered cigarette packs could be off-putting due to the absence of a familiar brand name, although the impact on smokers was considered negligible. Although adolescents were not clear on the rationale for numbered cigarette packs, some suggested that this would reduce one of the few remaining promotional features on standardised packs.

Challenges associated with e-cigarette use by people in custody in Scottish prisons: a qualitative interview study with prison staff.

OBJECTIVES: Little is known about the perspectives of staff working in prisons where e-cigarettes are permitted. Scotland now permits people in custody (PiC), but not staff/visitors to use e-cigarettes, following implementation of smoke-free prisons policy in 2018. Previous studies, conducted before and immediately after the introduction of e-cigarettes in Scottish prisons, have evidenced stakeholder support for their use by PiC. This study focuses on key challenges associated with e-cigarette use in prisons, using data collected from prison staff once e-cigarettes had been allowed in a smoke-free environment for 6-9 months. SETTING: Five prisons in Scotland. PARTICIPANTS: Sixteen qualitative interviews were conducted with prison staff from five prisons varying by population (sex, age and sentence length). Data were managed and analysed using the framework approach. RESULTS: While these staff confirmed strong support for the smoke-free prison policy and reported some benefits of replacing tobacco with e-cigarettes, they also spoke of the challenges e-cigarettes pose. These included: workplace e-cigarette vapour exposures; perceptions that e-cigarettes provide a new, effective way for some PiC to take illegal drugs, particularly new psychoactive substances; organisational challenges relating to the value attached to e-cigarettes in prisons; and implications for long-term nicotine use and tobacco cessation. Staff anticipated difficulties in tightening restrictions on e-cigarette use by PiC given its scale and significance among this population. CONCLUSIONS: Maximising the benefits of e-cigarette use by PiC is likely to require multiple measures to support effective and safe use and e-cigarette reduction/cessation where desired. This includes monitoring any misuse of e-cigarettes, and provision of guidance and support on appropriate e-cigarette use and how to limit or quit use if desired. Findings are relevant to jurisdictions considering or planning changes in prison smoking or vaping policies.

Post-implementation perspectives on smokefree prison policy: a qualitative study with staff and people in custody.

BACKGROUND: A comprehensive smokefree prison policy (SFPP) was introduced in Scottish prisons from November 2018, reflecting concern about inequalities in occupational exposures to second-hand smoke (SHS) and tobacco-related harms among people in custody (PiC). We aimed to address a gap, whereby few studies have sought to understand SFPP from the perspectives of people living and working in prisons. METHODS: As part of a comprehensive evaluation, focus groups and interviews with staff (n = 99) and interviews with PiC (n = 23) were conducted 6-9 months post-implementation of SFPP in Scotland. Data were analysed using the framework approach. RESULTS: Our study found that new restrictions on smoking had been widely accepted by PiC, after a period of adjustment which was less troublesome than participants had anticipated. Benefits of the SFPP for the safety and comfort of staff and PiC who were no longer exposed to SHS, and additionally for the health of PiC who were now smoking-abstinent, were widely acknowledged. Drawbacks of the SFPP, such as difficulties managing without tobacco and use of alternatives (e.g. e-cigarettes and changes in use of illegal drugs), were also reported. Contraband tobacco was not reported to be a major problem following prisons becoming smokefree. CONCLUSIONS: The findings strengthen evidence that SFPPs can be implemented without causing major disruption and highlight the need for removal of tobacco to be underpinned by careful planning, partnership working and ensuring the availability of support for smokers. Experiences from Scotland may be of interest and some comfort, internationally for jurisdictions considering smoke-free rules in prisons.

E-Cigarette Advertising in the UK: A Content Analysis of Traditional and Social Media Advertising to Observe Compliance with Current Regulations.

INTRODUCTION: The advertising of e-cigarettes in the UK is regulated through the revised EU Tobacco Products Directive and the Tobacco and Related Products Regulations, with further rules set out in the Advertising Standards Authority (ASA) Committees of Advertising (CAP) Code. Focusing on the ASA CAP Code Rules, we examined e-cigarette advertising regulation compliance in traditional advertising channels and on social media. METHODS: We conducted a content analysis of UK e-cigarette and related product advertising using a randomly selected sample (n = 130) of advertising in traditional channels and on Instagram which appeared between January and December 2019. All ads were independently double-coded to assess compliance with each CAP Code Rule. RESULTS: In traditional channels, our sample of advertising had largely good compliance. Only very small numbers of these ads appeared to be clearly in breach of any of the ASA rules (5% were in breach of Rule 22.7; 2% of Rule 22.9; and 1% of Rule 22.10). In contrast, we judged that all of the Instagram sample (n = 30) was in breach of Rule 22.12. For some rules, it was not possible to make definitive judgments about compliance, given uncertainty regarding how a rule should be interpreted and applied. CONCLUSIONS: We found overall good compliance for advertising in traditional channels, but assessed all of our social media advertising samples was in breach of regulations. Current guidance on e-cigarette advertising could be improved to facilitate e-cigarette advertising assessment and regulation. It would be beneficial to bring consumer perspectives into the assessment of regulation compliance. IMPLICATIONS: The regulation of e-cigarette advertising is a global concern. The UK Government has a statutory obligation to review the Tobacco and Related Products Regulations by May 2021. This study assessed compliance with current UK e-cigarette advertising regulations on placement and content. We identified areas where greater clarity is needed and outlined implications for future regulation.

The impact of the initial Covid-19 lockdown upon regular sports bettors in Britain: Findings from a cross-sectional online study.

BACKGROUND: In Britain, unprecedented restrictions on daily life associated with the Covid-19 pandemic included the suspension of professional sports events during the initial 'lockdown'. This provides opportunities to observe changes in sports bettors' behaviour when their primary form of activity is removed and assess the impact of Covid-19 related circumstances upon gambling. METHODS: In July 2020, we conducted an online cross-sectional survey of people who bet regularly (at least monthly) on sports before Covid-19 (n = 3866). Bi-variate analyses compared changes in gambling behaviours before and during the initial lockdown. Multi-variate logistic regression models explored associations between problem gambling (men) and moderate risk or problem gambling (MRPG) (women) with changes in Covid-19 related circumstances and changing gambling behaviours during Britain's initial 'lockdown' (March-June 2020). RESULTS: 29.8% of male sports bettors and 33.4% of female sports bettors stopped gambling altogether during the initial Covid-19 lockdown, though 17.3% of men and 16.5% of women started a new form of gambling during lockdown. Among men, adjusted odds ratios of problem gambling were higher among those starting a new gambling activity during lockdown (OR = 2.50 [95% CI 1.38-4.53]). Among women, adjusted odds ratios of MRPG were higher among those whose frequency of gambling on any activity increased during lockdown (OR = 4.21 [1.99-8.92] and among those shielding for health reasons. Poorer wellbeing was associated with problem gambling for men and MRPG for women. CONCLUSIONS: Those changing gambling behaviours during the initial Covid-19 lockdown (e.g. increasing gambling frequency or starting a new gambling activity) are potentially vulnerable to gambling harms.

Reactions to, and trial intentions for, three dissuasive cigarette designs: a cross-sectional survey of adolescents in Scotland.

OBJECTIVES: There has been growing academic and policy interest in opportunities to decrease the appeal of cigarette sticks, such as making them an unattractive colour or requiring them to display a health warning. We therefore explored reactions to, and trial intentions for, three 'dissuasive' cigarette designs among adolescents in Scotland. METHODS: A cross-sectional survey with 12-17 year olds in Scotland (n=594) was conducted between November 2017 and November 2018. Participants were shown one 'standard' cigarette (imitation cork filter with white paper casing) and three dissuasive cigarettes: (1) a cigarette with the warning 'smoking kills'; (2) a cigarette with the warning 'toxic' and a skull and cross-bones image and (3) a dark green cigarette. Participants rated each cigarette on nine five-point reaction measures (eg, appealing/unappealing or attractive/unattractive). A composite reaction score was computed for each cigarette, which was binary coded (overall negative reactions vs neutral/positive reactions). Participants also indicated whether they would try each cigarette (coded: Yes/No). Demographics, smoking status and smoking susceptibility were also measured. RESULTS: More participants had negative reactions to the dark green (93% of adolescents), 'smoking kills' (94%) and 'toxic' (96%) cigarettes, compared with the standard cigarette (85%). For all three dissuasive designs, Chi-square tests found that negative reactions were more likely among younger adolescents (vs older adolescents), never-smokers (vs ever smokers) and non-susceptible never-smokers (vs susceptible never-smokers). Most participants indicated that they would not try any of the cigarettes (range: 84%-91%). CONCLUSION: Dissuasive cigarettes present an opportunity to further reduce the appeal of smoking among adolescents.

E-cigarette Use in Prisons With Recently Established Smokefree Policies: A Qualitative Interview Study With People in Custody in Scotland.

INTRODUCTION: E-cigarettes were one measure introduced to help people in custody (PiC) to prepare for and cope with implementation of comprehensive smokefree policies in Scottish prisons. Our earlier study explored experiences of vaping when e-cigarettes were first introduced and most participants were dual tobacco and e-cigarette users. Here we present findings of a subsequent study of vaping among a different sample of PiC when use of tobacco was prohibited in prison, and smokefree policy had become the norm. METHODS: Twenty-eight qualitative interviews were conducted with PiC who were current or former users of e-cigarettes in prison, 6-10 months after implementation of a smokefree policy. Data were managed and analyzed using the framework approach. RESULTS: PiC reported that vaping helped with mandated smoking abstinence. However, findings suggest that some PiC may be susceptible to heavy e-cigarette use potentially as a consequence of high nicotine dependence and situational factors such as e-cigarette product choice and availability in prisons; issues with nicotine delivery; prison regimes; and use of e-cigarettes for managing negative emotions. These factors may act as barriers to cutting down or stopping use of e-cigarettes by PiC who want to make changes due to dissatisfaction with vaping or lack of interest in continued use of nicotine, cost, and/or health concerns. CONCLUSIONS: E-cigarettes helped PiC to cope with smokefree rules, although concerns about e-cigarette efficacy, cost, and safety were raised. PiC may desire or benefit both from conventional smoking cessation programs and from interventions to support reduction, or cessation, of vaping. IMPLICATIONS: Findings highlight successes, challenges, and potential solutions in respect of use of e-cigarettes to cope with mandated smoking abstinence in populations with high smoking prevalence and heavy nicotine dependence. Experiences from prisons in Scotland may be of particular interest to health and/or justice services in other jurisdictions, with similar legislation on e-cigarettes to the United Kingdom, who are planning for institutional smokefree policies in their prisons or inpatient mental health settings in the future.

Protocol for a Mixed-Method Investigation of the Impact of the COVID-19 Pandemic and Gambling Practices, Experiences and Marketing in the UK: The "Betting and Gaming COVID-19 Impact Study".

The COVID-19 pandemic led to unprecedented restrictions on people's movements and interactions, as well as the cancellation of major sports events and social activities, directly altering the gambling landscape. There is urgent need to provide regulators, policy makers and treatment providers with evidence on the patterns and context of gambling during COVID-19 and its aftermath. This protocol describes a study addressing the following three questions: (1) How has COVID-19 changed gambling practices and the risk factors for, and experience of, gambling harms? (2) What is the effect of COVID-19 on gambling marketing? (3) How has COVID-19 changed high risk groups' gambling experiences and practices? This mixed-method study focuses on two groups, namely young adults and sports bettors. In workpackage-1, we will extend an existing longitudinal survey of gambling in young adults (aged 16-24 years) (first wave conducted June-August 2019), adding COVID-19-related questions to the second wave (July-August 2020) and extending to a third wave in 2021; and undertake a survey of sports bettors in the UK (baseline n = 4000, ~July-August 2020), with follow-ups in ~October-November 2020 and ~February-March 2021. In workpackage-2, we will examine changes in expenditure on paid-for gambling advertising from January 2019 to July 2021 and undertake a mixed-method content analysis of a random sample of paid-for gambling advertising (n ~ 200) and social media marketing (n ~ 100) during the initial COVID-19 "lockdown". Workpackage-3 will involve qualitative interviews with a purposive sample of (a) young adults (aged 18-24 years) and (b) sports bettors.

Reactions to Standardized Cigarette Packs With Varying Structural Designs, and the Association With Smoking Susceptibility: A Postimplementation Cross-Sectional Survey With Never-Smoking Adolescents in Scotland.

INTRODUCTION: From May 20, 2017, cigarettes in the United Kingdom must be sold in standardized (plain) packaging. We explore postimplementation reactions to standardized cigarette packaging among never-smokers in Scotland, whether reactions vary in relation to permitted variations in pack structure, and whether reactions are associated with susceptibility. AIMS AND METHODS: A cross-sectional survey with 12-17-year-old never-smokers (n = 507) in Scotland, conducted November 2017-November 2018. Participants were shown one "regular" standardized cigarette pack (flip-top lid and straight-edged pack, similar to designs in Australia) and three standardized packs with varied pack structures (beveled-edges, slim pack, and shoulder box), which are permitted postimplementation in the United Kingdom. Participants rated each pack on eight five-point reaction measures (eg, attractiveness). Participants also indicated which pack, if any, they would choose. Smoking susceptibility was the outcome. RESULTS: The mean reaction scores for all four packs were mostly negative, however the shoulder box was consistently rated less negatively than the regular, slim, or beveled-edge packs. Most participants (87%) said they would not select any of the four packs, although susceptible participants were more likely to select one than nonsusceptible participants (25% vs. 7%; χ 2 = 29.70; p < .001). For all four packs, not finding them off-putting was associated with susceptibility (Adjusted Odds Ratio range: 2.73-3.69), albeit only a minority of adolescents did not find each pack off-putting. CONCLUSIONS: Adolescents have negative reactions to the standardized cigarette packs implemented in the United Kingdom, albeit permitted variations in structure can reduce the extent of negativity. Most reactions to standardized packaging had no association with susceptibility. IMPLICATIONS: We provide the first empirical evidence that adolescents find the standardized cigarette packs implemented in the United Kingdom unappealing and that most pack reactions have no association with susceptibility among never-smokers, with the exception of the minority who did not think that they would put them off smoking. This suggests that the legislation is achieving one of its primary aims, to reduce the appeal of packaging. That permitted variations in pack structure (eg, shoulder boxes) somewhat reduce negative reactions suggests that the United Kingdom, and other countries introducing similar legislation, should ensure that all aspects of pack design are fully standardized.

Smokers' Perceptions of Cigarette Packaging With Audio Warning Messages: A Focus Group Study in Scotland.

AIMS: The use of audio pack cues or messages is a recent trend in packaging design. There is scope to use audio technology to communicate health and cessation messages via cigarette packaging. We explored how smokers responded to cigarette packs which played audio health messages. METHODS: Twenty focus groups were conducted in Scotland in 2015 with smokers (n = 120) segmented by age (16-17, 18-24, 25-35, 36-50, >50), gender and social grade. Perceptions of cigarette packs which played audio warnings were explored, with four messages used: (1) a cessation message with a quitline number, (2) mortality message, (3) fertility message, and (4) message about ageing skin. RESULTS: Audio warnings were thought to increase message impact, particularly among younger smokers, as they would be hard to ignore or avoid, and repetition may lead to the messages being memorized. The warnings were considered annoying or embarrassing, and participants suggested they may discard the packs and use alternative storage. Some participants suggested that the audio warnings were off-putting and may alter their smoking behavior, with the mortality message deemed most effective and was considered relatable and felt personal. Older smokers were least likely to believe that audio warnings would affect their smoking behavior, although some thought that they may enhance cessation attempts among smokers seeking to quit, and could deter new or potential smokers, for example, young people. CONCLUSIONS: Cigarette packs with audio messaging may have a role to play, now or in the future, as a novel way of communicating health and cessation information. IMPLICATIONS: There is a lack of research exploring smokers' perceptions of cigarette packs which play audio health messages. This focus group study provides an understanding of smokers' immediate responses to cigarette packs which played a short health message when opened. Smokers generally viewed them as annoying or embarrassing, and some suggested the use of alternative storage. Audio warnings were thought to increase message salience, memorability, and impact, for younger smokers in particular, and some suggested that they be off-putting for themselves or others.

Adolescents' perceptions of standardised cigarette packaging design and brand variant name post-implementation: a focus group study in Scotland.

BACKGROUND: The United Kingdom (UK) fully-implemented standardised packaging for cigarettes and rolling tobacco on 20th May 2017. We explore adolescent's awareness of, and responses to, standardised cigarette packaging in the UK after it became mandatory. METHODS: Eight focus groups were conducted in schools in Scotland with 16-17 year-olds (n = 41), between November 2017 and November 2018, to explore awareness of, and responses to, standardised cigarette packaging. Unlike in Australia, where only straight-edged flip-top cigarette packs are permitted, in the UK standardised cigarette packs can have slim designs, and different edge types (straight, rounded or bevelled) and opening styles (flip-top or shoulder box). We explored how each of these pack formats was perceived. We also explored to what extent brand variant name differentiated cigarettes sold in standardised packaging. RESULTS: Most participants were aware of standardised packaging without being shown pack stimuli. Standardised packs were considered embarrassing and off-putting, and the health warnings salient. Among the standardised packs shown, there was a preference for the slimmer pack, viewed as more discrete and the cigarettes potentially less harmful, and the shoulder box, considered cool and different. Participants were interested in some brand variant names on standardised packs (e.g. Legendary Black), particularly those they considered to imply coolness and sophistication. CONCLUSION: Adolescents consider standardised cigarette packs in the UK unappealing, and the warnings salient, two core aims of this measure. However, positive reactions to some of the standardised packs (slimmer pack, shoulder box), and variant names used, has implications for countries developing standardised packaging regulations.

How tobacco companies in the United Kingdom prepared for, and responded to, standardised packaging of cigarettes and rolling tobacco.

INTRODUCTION: As a result of the Standardised Packaging of Tobacco Products Regulations and Tobacco Products Directive, all packs of cigarettes (factory-made and hand-rolled) in the UK must be drab brown, display pictorial warnings on the principal display areas and contain no less than 20 cigarettes or 30 g of tobacco. The legislation was phased in between May 2016 and May 2017. Our objective was to monitor pack, brand and product changes preimplementation and postimplementation. METHODS: Our surveillance of the cigarette market involved a review of the trade press, a monthly monitor of online supermarkets and regular visits to stores, from May 2015 to June 2017. RESULTS: Before standardised packaging there were changes to the pack graphics (eg, redesigned packs and limited editions) and pack structure (eg, resealable inner foil) and the issue of a number of reusable tins. After standardised packaging, changes included newer cigarette pack sizes for some brand variants (eg, 23 and 24 packs). Changes to the branding prestandardised packaging included brand extensions, and poststandardised packaging included brand and/or variant name change, often with the inclusion of colour descriptors and brand migrations. Product changes prestandardised packaging included the introduction of novel filters (eg, filters with two flavour-changing capsules, tube filters, firmer filters and filters with granular additives). There was non-compliance with the legislation, with slim packs, which are not permitted, on sale after standardised packaging was implemented. CONCLUSIONS: Our findings highlight the need to monitor developments in markets introducing standardised packaging and have policy implications for countries considering this measure.

The Use of Antibodies in Small-Molecule Drug Discovery.

Antibodies are powerful research tools that can be used in many areas of biology to probe, measure, and perturb various biological structures. Successful drug discovery is dependent on the correct identification of a target implicated in disease, coupled with the successful selection, optimization, and development of a candidate drug. Because of their specific binding characteristics, with regard to specificity, affinity, and avidity, coupled with their amenability to protein engineering, antibodies have become a key tool in drug discovery, enabling the quantification, localization, and modulation of proteins of interest. This review summarizes the application of antibodies and other protein affinity reagents as specific research tools within the drug discovery process.

Effect of the HIV protease inhibitor amprenavir on the growth and differentiation of primary gingival epithelium.

BACKGROUND: HIV-positive patients taking antiretroviral drugs, including protease inhibitors, have shown a significant increase in the development of oral complications; these complications are a major health issue for affected patients. The effect of these drugs on oral epithelium growth and differentiation is presently unknown. In this study, we explore for the first time the effect of the HIV protease inhibitor amprenavir on gingival epithelium growth and differentiation. METHODS: Organotypic (raft) cultures of gingival keratinocytes were established and the raft cultures treated with a range of amprenavir concentrations. Haematoxylin and eosin staining was performed to examine the effect of amprenavir on gingival epithelium growth and stratification. The raft cultures were also immunohistochemically analysed to determine the effect of amprenavir on the expression of key differentiation and proliferation markers, including cytokeratins, proliferating cell nuclear antigen (PCNA) and cyclin A. RESULTS: Amprenavir severely inhibited the growth of gingival epithelium when the drug was present throughout the growth period of the tissue. When the drug was added at day 8, amprenavir treatment altered the proliferation and differentiation of gingival keratinocytes. Expression of the cytokeratins 5, 14, 6 and 10, PCNA and cyclin A was increased; their expression pattern was also altered over time in treated rafts. Biochemically, the tissue exhibited characteristics of increased proliferation in the suprabasal layers of amprenavir-treated tissue. CONCLUSIONS: Our results suggest that amprenavir treatment deregulates the cell cycle/proliferation and differentiation pathways, resulting in abnormal epithelial repair and proliferation. Our system could be developed as a potential model for studying the effects of highly active antiretroviral therapy in vitro.

Sperm contamination in archived and contemporary herring samples.

Studies using archived scales and otoliths to examine ancient fish populations have become increasingly common, despite many methodological challenges in ancient DNA research. Here, we describe a case of DNA contamination in both modern and historical samples of Pacific herring (Clupea pallasii), where the source of the contamination is likely from milt spillage during collection. We describe a series of experiments to remove contamination using pre-extraction wash treatments. Though contamination was easily removed from contemporary fin clippings, no method was successful at removing contamination from historical scales. We discuss the implications of our findings to the genetic analysis of archived samples.