BackgroundMid-to-late adolescence is a critical period for initiation of alcohol and drug problems, which can be reduced by targeted brief motivational interventions. Web-based brief interventions have advantages in terms of acceptability and accessibility and have shown significant reductions of substance use among college students. However, the evidence is sparse among adolescents with at-risk use of alcohol and other drugs.ObjectiveThis study evaluated the effectiveness of a targeted and fully automated Web-based brief motivational intervention with no face-to-face components on substance use among adolescents screened for at-risk substance use in four European countries.MethodsIn an open-access, purely Web-based randomized controlled trial, a convenience sample of adolescents aged 16-18 years from Sweden, Germany, Belgium, and the Czech Republic was recruited using online and offline methods and screened online for at-risk substance use using the CRAFFT (Car, Relax, Alone, Forget, Friends, Trouble) screening instrument. Participants were randomized to a single session brief motivational intervention group or an assessment-only control group but not blinded. Primary outcome was differences in past month drinking measured by a self-reported AUDIT-C-based index score for drinking frequency, quantity, and frequency of binge drinking with measures collected online at baseline and after 3 months. Secondary outcomes were the AUDIT-C-based separate drinking indicators, illegal drug use, and polydrug use. All outcome analyses were conducted with and without Expectation Maximization (EM) imputation of missing follow-up data.ResultsIn total, 2673 adolescents were screened and 1449 (54.2%) participants were randomized to the intervention or control group. After 3 months, 211 adolescents (14.5%) provided follow-up data. Compared to the control group, results from linear mixed models revealed significant reductions in self-reported past-month drinking in favor of the intervention group in both the non-imputed (P=.010) and the EM-imputed sample (P=.022). Secondary analyses revealed a significant effect on drinking frequency (P=.037) and frequency of binge drinking (P=.044) in the non-imputation-based analyses and drinking quantity (P=.021) when missing data were imputed. Analyses for illegal drug use and polydrug use revealed no significant differences between the study groups (Ps>.05).ConclusionsAlthough the study is limited by a large drop-out, significant between-group effects for alcohol use indicate that targeted brief motivational intervention in a fully automated Web-based format can be effective to reduce drinking and lessen existing substance use service barriers for at-risk drinking European adolescents.Trial RegistrationInternational Standard Randomized Controlled Trial Registry: ISRCTN95538913; http://www.isrctn.com/ISRCTN95538913 (Archived by WebCite at http://www.webcitation.org/6XkuUEwBx)
BackgroundDuring the last few decades the use of club drugs (e.g., cocaine, amphetamines) has been of increased concern in nightlife settings. Traditionally, surveys have been used to estimate the use of club drugs, however, they mostly rely on self-reports which may not be accurate. Recent advances have allowed for readily accessible drug testing methods such as oral fluid drug testing. Nevertheless, research using oral fluid sampling to measure the frequency of drug use in the club environment is scarce. The objective of this study is to evaluate the feasibility of measuring the frequency of alcohol and drug use among Swedish clubbers using breath alcohol and oral fluid drug testing.MethodThe setting was a 40 hour electronic music dance event (EMDE) on a cruise ship on the Baltic Sea, departing from Sweden, with 875 passengers. Groups of participants at the EMDE were randomly invited to participate. Data were collected with face-to-face and self-administered questionnaires. Further, oral fluid samples were collected to determine illicit drug use, and blood alcohol concentration (BAC) levels were measured using a breath analyzer.ResultsA total of 422 passengers were asked to participate in the study whereof 21 declined (5.0% refusal rate). Of the 401 study participants (accounting for 45.8% of all attendees), 5 declined oral fluid drug testing. Results show that there was a discrepancy between self-reported and actual drug use as 10.1% of the participants were positive on illicit drug use (amphetamines, ecstasy/MDMA, cannabis, cocaine), while only 3.7% of the participants reported drug use during the last 48 hours. The average BAC level was 0.10% and 23.7% had BAC levels ≥ 0.15%, while 5.9% had levels below the detection limit. The mean BAC levels for the illicit drug users were significantly higher (p = 0.004) than for non-drug users (0.13% vs. 0.10%). Self-reported AUDIT-C scores (using a threshold of ≥ 5 for men and ≥ 4 for women) revealed that 76.0% of the men and 80.7% of the women had risky alcohol consumption patterns.ConclusionThis study indicates that it is feasible to conduct breath alcohol and oral fluid drug testing in a Swedish club setting.
Glutaredoxins (Grxs) are glutathione-dependent oxidoreductases that belong to the thioredoxin superfamily catalyzing thiol-disulfide exchange reactions via active site cysteine residues. Focusing on the human dithiol glutaredoxins having a C-X-Y-C active site sequence motif, the redox potentials of hGrx1 and hGrx2 were determined to be -232 and -221 mV, respectively, using a combination of redox buffers, protein-protein equilibrium and thermodynamic linkage. In addition, a nonactive site disulfide was identified between Cys28 and Cys113 in hGrx2 using redox buffers and chemical digestion. This disulfide confers nearly five kcal mol(-1) additional stability by linking the C-terminal helix to the bulk of the protein. The redox potential of this nonactive site disulfide was determined to be -317 mV and is thus expected to be present in all but the most reducing conditions in vivo. As all human glutaredoxins contain additional nonactive site cysteine residues, a full phylogenetic analysis was performed to help elucidate their structural and functional roles. Three distinct groups were found: Grx1, Grx2, and Grx5, the latter representing a highly conserved group of monothiol glutaredoxins having a C-G-F-S active site sequence, with clear homologs from bacteria to human. Grx1 and Grx2 diverged from a common ancestor before the origin of vertebrates, possibly even earlier in animal evolution. The highly stabilizing nonactive site disulfide observed in hGrx2 is found to be a conserved feature within the deuterostomes and appears to be the only additional conserved intramolecular disulfide within the glutaredoxins.
These results indicate that the problem is widespread. Our findings are similar to previous research where a more indirect methodology has been adopted, using either psychiatric interviews or self-reported alcohol consumption of adults, to estimate the magnitude of the problem.
Our results showed that a substantial number of children in Sweden have parents with a SUD and that it is important to consider both alcohol and drugs, when estimating the size of this group. Our findings call for further strategies to support these children and their families.
Background/Aims: Not enough is known about the psychometric properties of screening instruments for problematic alcohol consumption among adolescents. The aim of the current study was to evaluate and compare the performance of the screening instruments: Alcohol Use Disorders Identification Test (AUDIT), AUDIT-C, CRAFFT, and the alcohol domain of Alcohol, Smoking and Substance Involvement Screening Test-Youth (ASSIST-Y) among adolescents and to suggest optimal cut-offs indicating problematic use. Methods: Data was collected from a general population sample (n = 1,421) and a treatment-seeking sample (n = 59) using electronic versions of the instruments. Results: The internal consistencies for the instruments were fair (alpha’s AUDIT 0.74, AUDIT-C 0.75, CRAFFT 0.67, ASSIST-Y 0.62), and test-retest reliabilities were good to excellent (intraclass correlation coefficients AUDIT 0.86, AUDIT-C 0.93, CRAFFT 0.77, ASSIST-Y 0.63). The CRAFFT and ASSIST-Y demonstrated reasonable construct validities while factor solutions for AUDIT and AUDIT-C could not be determined. The optimal cut-off score was 2 for both CRAFFT and ASSIST-Y (61 and 73% sensitivities and 79 and 65% specificities, respectively), while sensitivity scores were poor for AUDIT and AUDIT-C. Conclusion: Based on the current sample, ASSIST-Y and the CRAFFT performed better than AUDIT and AUDIT-C. Health-care clinics working with adolescents should carefully consider their choice of screening instruments.
Traditionally, quantification of protein-ligand affinity is performed using kinetic or equilibrium measurements. However, if the binding reaction proceeds via a stable covalent complex, these approaches are often limited. By exploiting the fact that the conformational stabilization of a protein is altered upon ligand binding due to specific interactions, and using an array of selectively chosen ligand analogs, one can quantify the contribution individual interactions have on specificity. We have used ligand-induced stability as a basis to dissect the interaction between glutaredoxin-3 (Grx3) and one of its native substrates, the tripeptide glutathione. Taking advantage of the fact that Grx3 can be trapped in a covalent mixed disulfide to glutathione or to selected synthetic glutathione analogs as part of the natural catalytic cycle, individual contributions to binding of specific molecular groups can be quantified by changes in ligand-induced stability. These changes in conformational stability are interpreted in terms of interaction energies (i.e. specificity) of the particular groups present on the ligand analog. Our results illustrate that although Grx3 recognizes glutathione predominantly through independent and additive ionic interactions at the N-and C-terminal of glutathione, van der Waals interactions from the unique ␥-glutamate moiety of glutathione also play an important role. This study places us closer to understanding the complex task of accommodating multiple substrate specificities in proteins of the thioredoxin superfamily and underscores the general applicability of ligand-induced stability to probe substrate specificity.In the era of structural genomics, a large number of biomolecules will have their tertiary structure determined. However, to fully appreciate the potential of these structures, it is fundamental to determine how these molecules interact with other molecules. Therefore, when exploring the structural basis of observed biological activity it is of great importance to investigate the details of intermolecular interactions. One way of attaining a deeper insight into these interactions is by quantifying ligand affinity, usually performed by kinetic or equilibrium measurements of the binding reaction. Specific interactions between a ligand and a protein can be investigated further by modifying specific residues within the protein using site-directed mutagenesis. The relative binding constants of a wildtype and a mutant protein to a particular ligand then provide information about specificity. However, protein mutagenesis sometimes results in secondary effects as intrinsic properties of the protein may be altered, making subsequent analysis complex. Furthermore, if the interaction between two molecules involves a relatively stable covalent intermediate, standard methods of measuring affinity are often of limited value.Several studies have demonstrated that the free energy of non-covalent binding of a ligand to a protein results in a change in conformational stabilization with respect to...
Alcohol use and alcohol-related problems, including accidents, vandalism and violence, at sporting events are of increased concern in Sweden and other countries. The relationship between alcohol use and violence has been established and can be explained by the level of intoxication. Given the occurrence of alcohol use and alcohol-related problems at sporting events, research has assessed intoxication levels measured through biological sampling among spectators. This cross-sectional study aimed to assess the level of alcohol intoxication among spectators at football matches in the Swedish Premier Football League. Spectators were randomly selected and invited to participate in the study. Alcohol intoxication was measured with a breath analyser for Blood Alcohol Concentration levels, and data on gender, age, and recent alcohol use were gathered through a face-to-face interview. Blood Alcohol Concentration samples from 4420 spectators were collected. Almost half (46.8%) had a positive Blood Alcohol Concentration level, with a mean value of 0.063%, while 8.9% had a Blood Alcohol Concentration level ≥ 0.1%, with a mean value of 0.135%. Factors that predicted a higher Blood Alcohol Concentration level included male gender (p = 0.005), lower age (p < 0.001), attending a local derby (p < 0.001), alcohol use prior to having entered the arena (p < 0.001), attending a weekend match (p < 0.001), and being a spectator at supporter sections (p < 0.001). About half of all spectators at football matches in the Swedish Premier Football League drink alcohol in conjunction with the match. Approximately one tenth have a high level of alcohol intoxication.
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