Cell Line and Media Composition Influence the Production of Giant Plasma Membrane Vesicles.

Giant plasma membrane vesicles (GPMVs) have been utilized as a model to study phase separation in the plasma membrane. Additionally, GPMVs have been employed as vehicle for delivering molecular cargo, including small molecule drugs and nanoparticles. Nearly all examples of GPMV production use a defined salt buffer that is a stark contrast to typical cell culture medium. In this study, we demonstrate that the addition of formaldehyde and dithiothreitol to a standard culture medium was capable of generating GPMVs at a concentration equal to or higher than the traditional production buffer. These methods were evaluated for two human cell lines: kidney endothelial and Schwann cells (SCs). Morphological properties of the resultant GPMVs exhibited no significant differences between the two formulations. Factors such as pH and seeding density significantly influenced the production of GPMVs in both mediums. The cell type and seeding density was shown to influence the number of GPMVs to the greatest extent. SCs yield more GPMVs at higher seeding densities compared to endothelial cells. Stability of the membrane of the GPMVs produced in both mediums was evaluated by monitoring passive diffusion of two fluorescently tagged dextrans (3 and 10 kDa). Regardless of the production formulation or cell type, approximately 85% GPMVs are impermeable to either dextran. Cold storage for on-demand use and shipping are essential for broader use of GPMVs. Toward this aim, we have evaluated the GMPV number and morphologies following storage at -80 °C and in liquid nitrogen. A significant loss of the GPMV number, ∼30%, was observed following storage across production formulations as well as cell types. Our results indicate that smaller GMPVs, <5 µm are more stable for preservation. In conclusion, GPMVs can be produced in a broad range of formulations, exhibit a high degree of stability, and can undergo cold storage for further adoption.

Effects of alcohol intoxication on driving performance, confidence in driving ability, and psychomotor function: a randomized, double-blind, placebo-controlled study.

RATIONALE: Alcohol-induced driving impairment can occur with any departure from a zero-blood alcohol concentration (BAC). Because intoxication is characterised by impaired judgement, drivers under the influence of alcohol may overestimate their capacity to safely operate a vehicle. OBJECTIVES: This study examined the effects of alcohol on driving performance, four-choice reaction time (FCRT), and self-rated confidence in driving ability. It specifically focused on alcohol doses equal to commonly enforced legal BAC limits (i.e. 0.05% and 0.08%). METHODS: A randomized, double-blind, placebo-controlled design was utilised. Seventeen participants were tested in three conditions: placebo and two alcohol conditions aiming for BACs of 0.05% and 0.08%. Participants underwent a baseline FCRT task and a 1-h simulated highway driving task before completing another FCRT task and rated their confidence in their driving ability. RESULTS: The high and low alcohol dose conditions resulted in a mean BAC of 0.07%, and 0.04%, respectively (n = 17). The high BAC treatment significantly increased standard deviation of lateral position (SDLP) by 4.06 ± 5.21 cm and standard deviation of speed (SDS) by 0.69 ± 0.17 km/h relative to placebo, while confidence in driving ability remained unchanged across treatments. FCRT performance was impaired by the high BAC treatment (all < 0.01), but there we no significant differences between placebo and low BAC conditions. CONCLUSIONS: The findings of this study show that driving performance and associated psychomotor functioning become significantly impaired below legally permissible driving limits in some jurisdictions. We identified a dissociation between driving performance and subjective awareness of impairment. Despite a significantly diminished driving ability at 0.07% BAC, drivers were unaware of their impairment.

Effects of alcohol hangover on attentional resources during a verbal memory/psychomotor tracking dual attention task.

BACKGROUND: Alcohol hangover (AH) is associated with impaired attention and memory performance. However, whether this effect is related to reduced attentional resources remains unclear. AIMS: A dual-attention paradigm was employed to assess the effects of AH on attentional resources, delayed memory recognition, and the interaction between attentional load and AH. Mental effort and perceived performance during AH and control conditions were also assessed. METHODS: A seminaturalistic, crossover design was used. In total, 25 healthy social drinkers aged 18-35 years, visited the laboratory following a typical night out drinking (Hangover condition) and after alcohol abstinence (control) between 8:30 am and 12:30 pm, with conditions counterbalanced. Attentional load was manipulated via the presence (dual attention) or absence of psychomotor tracking during verbal memory encoding. Perceived mental effort and performance were measured using the NASA-TLX. Participants' recollected alcohol consumption was used to compute estimated blood alcohol level (eBAC). RESULTS: Compared with the control visit, AH was associated with reduced recognition accuracy (particularly more false negatives), higher "tracking costs" (poorer accuracy) in the dual attention condition, increased ratings of "mental demand," "effort," and "frustration," and lower ratings of task performance. There was also a significant main effect of attentional load with poorer recognition accuracy and response time in the dual attention condition. There were no significant interaction effects between hangover and attentional load. CONCLUSION: These findings suggest that reduced attentional resources contribute to the cognitive deficits associated with AH including impaired memory consolidation. They further suggest that while hungover, participants are aware of these deficits but are unable to compensate.

Associations between Mental Resilience, Mood, Coping, Personality, and Hangover Severity.

Extensive research exists on relationships between psychological constructs and alcohol consumption. However, research on relationships with hangover severity remains limited. This study aimed to assess the associations between mental resilience, mood (i.e., depression, anxiety, and stress), coping, personality, and hangover severity. A total of N = 690 participants completed an online survey by answering questions regarding their demographics, alcohol use, hangover prevalence and severity, and several psychological assessments (Brief Resilience Scale, DASS-21, Brief Cope, and Brief Version of the Big Five Personality Inventory). Significant associations were found between hangover severity and mental resilience, mood, and avoidant coping. Higher levels of mental resilience were associated with less severe hangovers, whereas poorer mood was associated with more severe hangovers. No significant associations were found with personality traits. These findings demonstrate that several associations between psychological constructs and hangover severity exist and suggest a role of psychological factors in the pathology of the alcohol hangover. As our findings contrast with the results of previous studies that did not report an association between mental resilience and the presence and severity of hangovers, further research is warranted.

Alcohol Consumption on the Heaviest Drinking Occasion and Hangovers during the First Dutch COVID-19 Lockdown.

The purpose of this study was to compare alcohol consumption between the heaviest drinking occasion in the period before the coronavirus disease 2019 (COVID-19) lockdown (15 January-14 March 2020) and the first COVID-19 lockdown period (15 March-11 May 2020) in the Netherlands, including the presence and severity of associated hangovers. The analysis included a sub-sample from the "Corona Lockdown: how fit are you?" (CLOFIT) study, comprising N = 761 participants who reported consuming alcohol in 2020. Overall, on the heaviest drinking occasion during the first COVID-19 lockdown period a significant reduction in number of alcoholic drinks consumed on the heaviest drinking occasion, drinking duration, and estimated BAC was observed. A significant reduction was also observed for subjective intoxication and next-day hangover severity. During the lockdown period, a significant reduction in the frequency of alcohol hangovers was reported. Several age and sex differences were observed. Specifically, men consumed significantly more alcohol than women and experienced hangovers significantly more frequently, both before and during the lockdown. With regard to age, young adults (18-35 years old) significantly reduced their alcohol intake on the heaviest drinking occasion during the lockdown and also reported lower ratings of subjective intoxication and hangover severity. No significant changes were seen for individuals above 35 years old. In conclusion, the first COVID-19 lockdown in the Netherlands was associated with reduced alcohol intake on the heaviest drinking occasion and a reduction in the severity of hangovers, particularly among young male adults.

Correction: Terpstra et al. Prevalence of Hangover Resistance According to Two Methods for Calculating Estimated Blood Alcohol Concentration (eBAC). J. Clin. Med. 2020, 9, 2823.

The authors wish to make the following corrections to the original article [...].

The Relationship between Alcohol Hangover Severity, Sleep and Cognitive Performance; a Naturalistic Study.

Alcohol hangover (AH) has been associated with poor sleep due to the negative effects of alcohol intoxication on sleep quantity and sleep quality. The aim of the current study was to further explore the relationship between AH severity and sleep using a naturalistic study design. A further aim was to determine whether quantitative aspects of sleep were a mediating influence on the relationship between AH severity and cognitive performance. As part of the naturalistic study design, 99 drinkers were recruited following a night of drinking in an Australian state capital, with breath alcohol concentration (BrAC) measured as participants were leaving the entertainment district. The following morning at home, participants answered online questions regarding their drinking behaviour on the previous evening, current AH symptoms and sleep quality. Participants also completed an online version of the Trail-Making Test B (TMT-B) to assess cognitive performance. The findings reveal the duration of nightly awakenings to be negatively related to six individual AH symptoms as well as overall AH severity. The number of nightly awakenings, sleep quality and total sleep time correlated with four AH symptoms including overall AH severity. Total AH severity accounted for a moderate amount of variance (11%) in the time to complete the TMT-B. These findings confirm that alcohol consumption negatively affects sleep, which is related to higher next-day hangover severity ratings and poorer cognitive performance.

Mood and Changes in Alcohol Consumption in Young Adults during COVID-19 Lockdown: A Model Explaining Associations with Perceived Immune Fitness and Experiencing COVID-19 Symptoms.

The aim of this study was to explore the effects of the 2019 Coronavirus (COVID-19) lockdown on mood states, stress, alcohol consumption and perceived immune fitness in a Dutch sample. Analysis included a subsample from the "Corona Lockdown: how fit are you?" (CLOFIT) study, comprising N = 761 participants who reported consuming alcohol in 2020. Results show that, compared to pre-lockdown, the first COVID-19 lockdown (March 2020) was associated with experiencing poorer mood (e.g., anxiety, depression, loneliness, fatigue) and increased stress levels. Among younger participants (18 to 35 years old), a significant decrease in weekly alcohol consumption was found during COVID-19 lockdown, which was not significant in older individuals. For the younger age cohort (18 to 35 years old), increased stress significantly correlated to increased weekly alcohol consumption (r = 0.163, p = 0.003), which in turn, correlated significantly to reporting a poorer perceived immune fitness (r = -0.165, p = 0.002). Poorer perceived immune fitness correlated significantly with increases in the presence and severity of COVID-19 symptoms (r = -0.313, p < 0.001, and r = -0.325, p < 0.001, respectively). The data provides evidence for significant relationships between changes in mood, stress and alcohol consumption during COVID-19 lockdown, and supports a model that links these changes to perceived immune fitness and susceptibility to experiencing COVID-19 symptoms.

Immune Fitness and the Psychosocial and Health Consequences of the COVID-19 Pandemic Lockdown in The Netherlands: Methodology and Design of the CLOFIT Study.

This article provides an overview of the design and methodology of the "Corona lockdown: how fit are you?" (CLOFIT) study, including the questionnaires and scales that were included in the online survey. The aim of the CLOFIT study was to investigate the psychosocial and health consequences of the coronavirus disease 2019 (COVID-19) pandemic in the Netherlands. The survey was conducted among the Dutch population to collect data on immune fitness and the psychological and health consequences of the 2019 coronavirus disease (COVID-19) pandemic lockdown in the Netherlands. The CLOFIT dataset contains measures from N = 1910 participants and is broadly representative of the Dutch general population. The dataset represents both sexes, a range of ages including the elderly, different education levels, and ethnic backgrounds. The cohort also includes people with a diverse health status and range of medication use.

A Cross-Cultural Comparison of the Effects of Alcohol Mixed with Energy Drink (AMED) Consumption on Overall Alcohol Consumption and Related Consequences.

There is a growing body of scientific literature examining the effects of alcohol mixed with energy drink (AMED) on alcohol consumption and related negative consequences, such as risky behavior or negative health effects. It is unknown whether differences in cultural context may influence these AMED effects. The current cross-cultural study compared the data of N = 6881 students from The Netherlands (N = 4424), UK (N = 1594), and Australia (N = 863). Demographics, alcohol consumption, its consequences, and motives for AMED consumption were assessed. Analyses included (a) between-groups comparison of AMED and alcohol only (AO) consumers, (b) within-subjects comparison of AMED and AO occasions among AMED consumers only, and (c) comparisons between the three countries. The between-groups analysis revealed that AMED consumers drink more alcohol than AO consumers (p < 0.001). AMED consumers differed from AO consumers in many other aspects. For example, AMED consumers were significantly more often a smoker and had higher risk-taking scores. Within subject analysis among AMED consumers showed that significantly less alcohol was consumed on AMED, compared to AO occasions (p < 0.001). These findings were observed for both typical drinking occasions and the past month's heaviest drinking occasion, and were consistent across the three countries. Comparisons between countries revealed that on both AMED and AO occasions, the UK sample consumed significantly more alcohol than the Australian and Dutch samples. Across countries, neutral motives such as 'I like the taste' and 'I wanted to drink something else' were the most frequently reported motives for consuming AMED. The most notable difference between the countries was the finding that consuming AMED 'To get drunk' was endorsed significantly more often among the UK sample (45.6%) than the Australian (31.2%) and Dutch (8.0%) samples. Negative alcohol-related consequences were significantly less frequently reported for AMED occasions compared to AO occasions. Some country-specific consequences of AMED consumption were observed, but these were more likely related to characteristics of the country and their drinking culture (e.g., the Australian sample reported more often driving a car after AMED consumption compared to the other countries, and this pattern of results was also found for AO occasions). In conclusion, there were limited differences between countries with regard to demographics of consumers and motives for AMED consumption, but the UK sample consumed significantly more alcohol and reported the highest frequency of negative alcohol related consequences. Consistent across countries was the observation that AMED consumers consume significantly less alcohol on their past month heaviest AMED drinking occasion, compared to their past month heaviest AO drinking occasion.

Risk-Taking Behavior and the Consumption of Alcohol Mixed with Energy Drink among Australian, Dutch and UK Students.

The relationship between risk-taking behavior, alcohol consumption and negative alcohol-related consequences is well known. The current analyses were conducted to investigate whether alcohol mixed with energy drink (AMED) is related to risk-taking behavior and if there is a relationship between the amount of energy drink mixed with alcohol consumed, risk-taking behavior and negative alcohol-related consequences. Data from N = 1276 AMED consuming students from the Netherlands, UK and Australia who completed the same survey were evaluated. The analysis revealed that, compared to AMED occasions, on alcohol only (AO) occasions significantly more alcohol was consumed and significantly more negative alcohol-related consequences were reported. On both AO and AMED occasions, there was a strong and positive relationship between amount of alcohol consumed, level of risk-taking behavior and number of reported negative alcohol-related consequences. In contrast, the level of risk-taking behavior was not clearly related to energy drink consumption. Across risk-taking levels, differences in the amount of energy drink consumed on AMED occasions did not exceed one 250 mL serving of energy drink. When correcting for the amount of alcohol consumed, there were no statistically significant differences in the number of energy drinks consumed on AMED occasions between the risk-taking groups. In conclusion, alcohol consumption is clearly related to risk-taking behavior and experiencing negative alcohol-related consequences. In contrast, energy drink intake was not related to level of risk-taking behavior and only weakly related to the number of experienced negative alcohol-related consequences.

Alcohol Hangover Across the Lifespan: Impact Of Sex and Age.

AIM: To investigate the relationship between age and hangover frequency and severity. METHOD: An online survey, generated through Facebook, collected self-report data relating to alcohol consumption from 761 Dutch alcohol consumers aged 18-94 years (61.6% female). RESULTS: Overall, young individuals consumed more alcohol than older drinkers, and men more than women. Significant interactions between age group and sex were found for both subjective intoxication and hangover severity, indicating that the sex differences in these variables were greatest in the younger age groups but became significantly smaller or absent in the older age groups. Partial correlations, correcting for estimated blood alcohol concentration (eBAC), revealed significant and negative partial correlations between age and subjective intoxication (r = -0.444, P < 0.0001), age and hangover severity (r = -0.327, P < 0.0001) and between age and hangover frequency (r = -0.195, P < 0.0001), i.e. subjective intoxication, hangover severity and hangover frequency decline with age. With regard to sex differences, the observed correlations with age for the past month heaviest drinking occasion were stronger in men for subjective intoxication, (z = -2.25, P = 0.024), hangover severity (z = -3.36, P = 0.0008) and hangover frequency (z = -3.63, P = 0.0003). CONCLUSIONS: Hangover severity declines with age, even after controlling for eBAC or the amount of alcohol consumed. Sex differences were greatest in the younger age groups but became significantly smaller or absent in the older age groups. The relationship between age and hangover severity is strongly mediated by subjective intoxication. Pain sensitivity, lower with aging, might be a mediator.

The effects of alcohol intoxication on cognitive functions critical for driving: A systematic review.

Alcohol is the most frequently detected substance in drivers involved in road traffic collisions. Given that up to 35% of fatal road collisions are alcohol-related, it is important to determine the influence of alcohol intoxication on driving-related skills. This review provides an updated and systematic evaluation of the available research concerning the effect of alcohol intoxication on cognitive functions critical for driving. Databases EBSCOhost, PsycInfo, PubMed, Scopus, Transport Research International Documentation (TRID) and Web of Science were searched for controlled trials examining the effect of alcohol on divided attention, executive functioning, perception, psychomotor skills, reaction time and/or vigilance. Fourteen studies met the inclusion criteria and were included in this review. We found that each of the cognitive domains assessed in this review showed impairment at blood alcohol concentrations equal to or below the legal driving limit in many jurisdictions. Future research could determine the effects of alcohol on cognitive functioning with greater accuracy by employing more consistent, reliable and comparable measures while considering the translation of deficits to real-life driving.

The Alcohol Hangover Research Group: Ten Years of Progress in Research on the Causes, Consequences, and Treatment of the Alcohol Hangover.

The alcohol hangover is defined as the combination of negative mental and physical symptoms, which can be experienced after a single episode of alcohol consumption, starting when blood alcohol concentration (BAC) approaches zero. Here, we present the book "The alcohol hangover: causes, consequences, and treatment", written to celebrate the 10th anniversary of the Alcohol Hangover Research Group (AHRG), summarizing recent advances in the field of alcohol hangover research.

Prevalence of Hangover Resistance According to Two Methods for Calculating Estimated Blood Alcohol Concentration (eBAC).

Hangover resistance may be linked to an increased risk of continuing harmful drinking behaviours as well as involvement in potentially dangerous daily activities such as driving while hungover, mainly due to the absence of negative consequences (i.e., hangover symptoms) the day after alcohol consumption. The aim of this study was to examine the occurrence of claimed alcohol hangover resistance relative to estimated blood alcohol concentration (eBAC). A total of 1198 participants completed an online survey by answering questions regarding their demographics, alcohol consumption and occurrence of hangover. Two methods were used to calculate eBAC, one based on the modified Widmark Equation (N = 955) and the other from an equation averaging the total body water (TBW) estimates of Forrest, Watson, Seidl, Widmark and Ulrich (males only) (N = 942). The percentage of participants who claimed to be hangover resistant decreased rapidly with increasing eBAC and only a small number of hangover resistant drinkers remained at higher eBACs. Comparisons of the eBACs calculated by the two methods revealed significantly higher BACs when using the modified Widmark equation. These findings suggest that additional research for eBAC calculations is needed to improve accuracy and comprehensiveness of these equations for future alcohol hangover research.

The Impact of Mood and Subjective Intoxication on Hangover Severity.

The aim of this study was to investigate whether baseline mood and/or mood while drinking have an impact on alcohol hangover severity. A survey was held among N = 331 young adults (mean age = 23.6 years, range = 18-35 years). Demographics, alcohol consumption, subjective intoxication, and hangover severity were assessed for the past three days. In addition, mood (baseline, while drinking, and during hangover) was also assessed. N = 143 participants reported to be hungover on the day of assessment, N = 122 participants reported to have been hungover the previous day ('yesterday'), and N = 87 participants reported to have been hungover two days before the assessment ('2 days ago'). The analyses revealed that baseline mood and mood while drinking had no relevant effect on the amount of consumed alcohol and did not significantly contribute to hangover severity. However, hangover severity was associated with significantly increased negative affect, particularly with higher levels of subjective stress on the day of the hangover.

Effects of Rapid Recovery on Alcohol Hangover Severity: A Double-Blind, Placebo-Controlled, Randomized, Balanced Crossover Trial.

The aim of this study was to evaluate the efficacy of putative hangover treatment, Rapid Recovery, in mitigating alcohol hangover (AH) symptom severity. Using a double-blind, randomized, placebo-controlled, balanced crossover design, 20 participants attended the laboratory for two evenings of alcohol consumption, each followed by morning assessments of AH severity. Participants were administered Rapid Recovery and placebo on separate visits. In the first testing visit, participants self-administered alcoholic beverages of their choice, to a maximum of 1.3 g/kg alcohol. Drinking patterns were recorded and replicated in the second evening testing visit. In the morning visits, AH severity was assessed using questionnaires measuring AH symptom severity and sleep quality, computerized assessments of cognitive functioning as well as levels of blood biomarkers of liver function (gamma-glutamyl transferase (GGT)) and inflammation (high-sensitive C-reactive protein (hs-CRP)). There were no differences in the blood alcohol concentrations (BAC) obtained in the Rapid Recovery (mean = 0.096%) and placebo (mean = 0.097%) conditions. Participants reported significantly greater sleep problems in the Rapid Recovery compared to placebo condition, although this difference was no longer significant following Bonferroni's correction. There were no other significant differences between Rapid Recovery and placebo. These data suggest that Rapid Recovery has no significant effect on alcohol hangover nor on associated biomarkers.

Alcohol Hangover and Multitasking: Effects on Mood, Cognitive Performance, Stress Reactivity, and Perceived Effort.

The aim of this study was to examine the effects of hangover on mood, multitasking ability, and psychological stress reactivity to cognitive demand. Using a crossover design and semi-naturalistic methodology, 25 participants attended the laboratory in the morning following a night of (i) alcohol abstinence and (ii) alcohol self-administration during a typical night out (with order counterbalanced across participants). They completed a four-module multitasking framework (MTF, a widely used laboratory stressor) and a battery of questionnaires assessing mood, hangover symptom severity, and previous night's sleep. The effects of the MTF on mood and perceived workload were also assessed. Participants in the hangover condition reported significantly lower alertness and contentment coupled with a higher mental fatigue and anxiety. Multitasking ability was also significantly impaired in the hangover condition. Completion of the cognitive stressor increased reported levels of mental demand, effort, and frustration, and decreased perceived level of performance. MTF completion did not differentially affect mood. Lastly, participants rated their sleep as significantly worse during the night prior to the hangover compared with the control condition. These findings confirm the negative cognitive and mood effects of hangover on mood. They also demonstrate that hangover is associated with greater perceived effort during task performance.