Native folding of aggregation-prone recombinant proteins in Escherichia coli by osmolytes, plasmid- or benzyl alcohol-overexpressed molecular chaperones.

When massively expressed in bacteria, recombinant proteins often tend to misfold and accumulate as soluble and insoluble nonfunctional aggregates. A general strategy to improve the native folding of recombinant proteins is to increase the cellular concentration of viscous organic compounds, termed osmolytes, or of molecular chaperones that can prevent aggregation and can actively scavenge and convert aggregates into natively refoldable species. In this study, metal affinity purification (immobilized metal ion affinity chromatography [IMAC]), confirmed by resistance to trypsin digestion, was used to distinguish soluble aggregates from soluble nativelike proteins. Salt-induced accumulation of osmolytes during induced protein synthesis significantly improved IMAC yields of folding-recalcitrant proteins. Yet, the highest yields were obtained with cells coexpressing plasmid-encoded molecular chaperones DnaK-DnaJ-GrpE, ClpB, GroEL-GroES, and IbpA/B. Addition of the membrane fluidizer heat shock-inducer benzyl alcohol (BA) to the bacterial medium resulted in similar high yields as with plasmid-mediated chaperone coexpression. Our results suggest that simple BA-mediated induction of endogenous chaperones can substitute for the more demanding approach of chaperone coexpression. Combined strategies of osmolyte-induced native folding with heat-, BA-, or plasmid-induced chaperone coexpression can be thought to optimize yields of natively folded recombinant proteins in bacteria, for research and biotechnological purposes.

Consommation d'alcool et de benzodiozépines au troisième âge [Consumption of alcohol and benzodiazepines in the elderly]

In Switzerland, 9.1% of the general population accept regular consumption of BZD. In 65-74 years age group, 2% of cases display a high-risk alcohol consumption. Moderate risk consumption is present in 5% of cases. For the BZD, the cognitive difficulties settle in an insidious way; for alcohol, the daily consumption of fairly high quantities may lead to cognitive deterioration. At early stages, alcohol abusers show preserved neuropsychological performances. Gradually, the deficits will affect the remaining cognitive functions and become irreversible. This review indicates that the chronic consumption of alcohol and BZD in old age is at the origin of major clinical difficulties that need ad hoc training both for psychiatrists and general practitioners.

Alcohol-attributable mortality in Switzerland in 2011--age-specific causes of death and impact of heavy versus non-heavy drinking.

BACKGROUND: Alcohol use causes high burden of disease and injury globally. Switzerland has a high consumption of alcohol, almost twice the global average. Alcohol-attributable deaths and years of life lost in Switzerland were estimated by age and sex for the year 2011. Additionally, the impact of heavy drinking (40+grams/day for women and 60+g/day for men) was estimated. METHODS: Alcohol consumption estimates were based on the Addiction Monitoring in Switzerland study and were adjusted to per capita consumption based on sales data. Mortality data were taken from the Swiss mortality register. Methodology of the Comparative Risk Assessment for alcohol was used to estimate alcohol-attributable fractions. RESULTS: Alcohol use caused 1,600 (95% CI: 1,472 - 1,728) net deaths (1,768 deaths caused, 168 deaths prevented) among 15 to 74 year olds, corresponding to 8.7% of all deaths (men: 1,181 deaths; women: 419 deaths). Overall, 42,627 years of life (9.7%, 95% CI: 40,245 - 45,008) were lost due to alcohol. Main causes of alcohol-attributable mortality were injuries at younger ages (15-34 years), with increasing age digestive diseases (mainly liver cirrhosis) and cancers (particularly breast cancers among women). The majority (62%) of all alcohol-attributable deaths was caused by chronic heavy drinking (men: 67%; women: 48 %). CONCLUSION: Alcohol is a major cause of premature mortality in Switzerland. Its impact, among young people mainly via injuries, among men mainly through heavy drinking, calls for a mix of preventive actions targeting chronic heavy drinking, binge drinking and mean consumption.

Cannabis Dependence in Swiss Adolescents Exploration of the Role of Anxiety, Coping Styles, and Psychosocial Difficulties

This naturalistic cross-sectional study explores how and to what extent cannabis dependence was associated with intrapersonal aspects (anxiety, coping styles) and interpersonal aspects of adolescent functioning (school status, family relationships, peer relationships, social life). A convenience sample of 110 adolescents (aged 12 to 19) was recruited and subdivided into two groups (38 with a cannabis dependence and 72 nondependent) according to DSM-IV-TR criteria for cannabis dependence. Participants completed the State-Trait Anxiety Inventory (STAI-Y), the Coping Across Situations Questionnaire (CASQ), and the Adolescent Drug Abuse Diagnosis (ADAD) interview investigating psychosocial and interpersonal problems in an adolescent's life. Factors associated with cannabis dependence were explored with logistic regression analyses. The results indicated that severity of problems in social life and peer relationships (OR = 1.68, 95% CI = 1.21 - 2.33) and avoidant coping (OR = 4.22, 95% CI = 1.01 - 17.73) were the only discriminatory factors for cannabis dependence. This model correctly classified 84.5% of the adolescents. These findings are partially consistent with the "self-medication hypothesis" and underlined the importance of peer relationships and dysfunctional coping strategies in cannabis dependence in adolescence. Limitations of the study and implications for clinical work with adolescents are discussed.

Economics of dialysis dependence following renal replacement therapy for critically ill acute kidney injury patients.

BACKGROUND: The obective of this study was to perform a cost-effectiveness analysis comparing intermittent with continuous renal replacement therapy (IRRT versus CRRT) as initial therapy for acute kidney injury (AKI) in the intensive care unit (ICU). METHODS: Assuming some patients would potentially be eligible for either modality, we modeled life year gained, the quality-adjusted life years (QALYs) and healthcare costs for a cohort of 1000 IRRT patients and a cohort of 1000 CRRT patients. We used a 1-year, 5-year and a lifetime horizon. A Markov model with two health states for AKI survivors was designed: dialysis dependence and dialysis independence. We applied Weibull regression from published estimates to fit survival curves for CRRT and IRRT patients and to fit the proportion of dialysis dependence among CRRT and IRRT survivors. We then applied a risk ratio reported in a large retrospective cohort study to the fitted CRRT estimates in order to determine the proportion of dialysis dependence for IRRT survivors. We conducted sensitivity analyses based on a range of differences for daily implementation cost between CRRT and IRRT (base case: CRRT day $632 more expensive than IRRT day; range from $200 to $1000) and a range of risk ratios for dialysis dependence for CRRT as compared with IRRT (from 0.65 to 0.95; base case: 0.80). RESULTS: Continuous renal replacement therapy was associated with a marginally greater gain in QALY as compared with IRRT (1.093 versus 1.078). Despite higher upfront costs for CRRT in the ICU ($4046 for CRRT versus $1423 for IRRT in average), the 5-year total cost including the cost of dialysis dependence was lower for CRRT ($37 780 for CRRT versus $39 448 for IRRT on average). The base case incremental cost-effectiveness analysis showed that CRRT dominated IRRT. This dominance was confirmed by extensive sensitivity analysis. CONCLUSIONS: Initial CRRT is cost-effective compared with initial IRRT by reducing the rate of long-term dialysis dependence among critically ill AKI survivors.

Parents' alcohol use: gender differences in the impact of household and family chores.

BACKGROUND: Social roles influence alcohol use. Nevertheless, little is known about how specific aspects of a given role, here parenthood, may influence alcohol use. The research questions for this study were the following: (i) are family-related indicators (FRI) linked to the alcohol use of mothers and fathers? and (ii) does the level of employment, i.e. full-time, part-time employment or unemployment, moderate the relationship between FRI and parental alcohol use? METHODS: Survey data of 3217 parents aged 25-50 living in Switzerland. Mean comparisons and multiple regression models of annual frequency of drinking and risky single occasion drinking, quantity per day on FRI (age of the youngest child, number of children in the household, majority of child-care/household duties). RESULTS: Protective relationships between FRI and alcohol use were observed among mothers. In contrast, among fathers, detrimental associations between FRI and alcohol use were observed. Whereas maternal responsibilities in general had a protective effect on alcohol use, the number of children had a detrimental impact on the quantity of alcohol consumed per day when mothers were in paid employment. Among fathers, the correlations between age of the youngest child, number of children and frequency of drinking was moderated by the level of paid employment. CONCLUSION: The study showed that in Switzerland, a systematic negative relationship was more often found between FRI and women's drinking than men's. Evidence was found that maternal responsibilities per se may protect from alcohol use but can turn into a detrimental triangle if mothers are additionally in paid employment.

Measuring alcohol-related consequences in school surveys: alcohol-attributable consequences or consequences with students' alcohol attribution.

In alcohol epidemiology surveys, there is a tradition of measuring alcohol-related consequences using respondents' attribution of alcohol as the cause. The authors aimed to compare the prevalence and frequency of self-attributed consequences to consequences without self-attribution using alcohol-attributable fractions (AAF). In 2007, a total of 7,174 Swiss school students aged 13-16 years reported the numbers of 6 alcohol-related adverse consequences (e.g., fights, injuries) they had incurred in the past 12 months. Consequences were measured with and without attribution of alcohol as the cause. The alcohol-use measures were frequency and volume of drinking in the past 12 months and number of risky single-occasion (> or =5 drinks) drinking episodes in the past 30 days. Attributable fractions were derived from logistic (> or =1 incident) and Poisson (number of incidents) regression analyses. Although relative risk estimates were higher when alcohol-attributed consequences were compared with nonattributed consequences, the use of AAFs resulted in more alcohol-related consequences (10,422 self-attributed consequences vs. 24,520 nonattributed consequences determined by means of AAFs). The likelihood of underreporting was higher among drinkers with intermediate frequencies than among either rare drinkers or frequent drinkers. Therefore, the extent of alcohol-related adverse consequences among adolescents may be underestimated when using self-attributed consequences, because of differential attribution processes, especially among infrequent drinkers.

Consommation de cannabis : quels sont les risques ?

Abstract - Cannabis: what are the risks ? Cannabinoids from cannabis have a dual use and display often opposite pharmacological properties depending on the circumstances of use and the administered dose. Cannabinoids constitute mainly a recreative or addictive substance, but also a therapeutic drug. They can be either neurotoxic or neuroprotector, carcinogenic or an anti-cancer drug, hyperemetic or antiemetic, pro-inflammatory or anti-inflammatory... Improvement in in-door cultivation techniques and selection of high yield strains have resulted in a steadily increase of THC content. Cannabis is the most frequently prohibited drug used in Switzerland and Western countries. About half of teenagers have already experimented cannabis consumption. About 10% of cannabis users smoke it daily and can be considered as cannabis-dependant. About one third of these cannabis smokers are chronically intoxicated. THC, the main psychoactive drug interacts with the endocannnabinoid system which is made of cellular receptors, endogenous ligands and a complex intra-cellular biosynthetic, degradation and intra-cellular messengers machinery. The endocannabinoid system plays a major role in the fine tuning of the nervous system. It is thought to be important in memory, motor learning, and synaptic plasticity. At psychoactive dose, THC impairs psychomotor and neurocognitive performances. Learning and memory abilities are diminished. The risk to be responsible of a traffic car accident is slightly increased after administration of cannabis alone and strongly increased after combined use of alcohol and cannabis. With the exception of young children, cannabis intake does not lead to potentially fatal intoxication. However, cannabis exposure can act as trigger for cardiovascular accidents in rare vulnerable people. Young or vulnerable people are more at risk to develop a psychosis at adulthood and/or to become cannabis-dependant. Epidemiological studies have shown that the risk to develop a schizophrenia at adulthood is increased for cannabis smokers, especially for those who are early consumers. Likewise for the risk of depression and suicide attempt. Respiratory disease can be worsen after cannabis smoking. Pregnant and breast-feeding mothers should not take cannabis because THC gets into placenta and concentrates in breast milk. The most sensitive time-period to adverse side-effects of cannabis starts from foetus and extends to adolescence. The reason could be that the endocannabinoid system, the main target of THC, plays a major role in the setup of neuronal networks in the immature brain. The concomitant use of other psychoactive drugs such as alcohol, benzodiazepines or cocaine should be avoided because of possible mutual interactions. Furthermore, it has been demonstrated that a cross-sensitisation exists between most addictive drugs at the level of the brain reward system. Chronic use of cannabis leads to tolerance and withdrawals symptoms in case of cannabis intake interruption. Apart from the aforementioned unwanted side effects, cannabis displays useful and original medicinal properties which are currently under scientific evaluation. At the moment the benefit/risk ratio is not yet well assessed. Several minor phytocannabinoids or synthetic cannabinoids devoid of psychoactive properties could find their way in the modern pharmacopoeia (e.g. ajulemic acid). For therapeutic purposes, special cannabis varieties with unique cannabinoids composition (e.g. a high cannabidiol content) are preferred over those which are currently used for recreative smoking. The administration mode also differs in such a way that inhalation of carcinogenic pyrolytic compounds resulting from cannabis smoking is avoided. This can be achieved by inhaling cannabis vapors at low temperature with a vaporizer device. Résumé Les cannabinoïdes contenus dans la plante de cannabis ont un double usage et possèdent des propriétés opposées suivant les circonstances et les doses employées. Les cannabinoïdes, essentiellement drogue récréative ou d'abus pourraient, pour certains d'entre eux, devenir des médicaments. Selon les conditions d'utilisation, ils peuvent être neurotoxiques ou neuroprotecteurs, carcinogènes ou anticancéreux, hyper-émétiques ou antiémétiques, pro-inflammatoires ou anti-inflammatoires... Les techniques de culture sous serre indoor ainsi que la sélection de variétés de cannabis à fort potentiel de production ont conduit à un accroissement notable des taux de THC. Le cannabis est la drogue illégale la plus fréquemment consommée en Suisse et ailleurs dans le monde occidental. Environ la moitié des jeunes ont déjà expérimenté le cannabis. Environ 10 % des consommateurs le fument quotidiennement et en sont devenus dépendants. Un tiers de ces usagers peut être considéré comme chroniquement intoxiqué. Le THC, la principale substance psychoactive du cannabis, interagit avec le "système endocannabinoïde". Ce système est composé de récepteurs cellulaires, de ligands endogènes et d'un dispositif complexe de synthèse, de dégradation, de régulation et de messagers intra-cellulaires. Le système endocannabinoïde joue un rôle clé dans le réglage fin du système nerveux. Les endocannabinoïdes régulent la mémorisation, l'apprentissage moteur et la plasticité des liaisons nerveuses. À dose psychoactive, le THC réduit les performances psychomotrices et neurocognitives. Les facultés d'apprentissage et de mémorisation sont diminuées. Le risque d'être responsable d'un accident de circulation est augmenté après prise de cannabis, et ceci d'autant plus que de l'alcool aura été consommé parallèlement. À l'exception des jeunes enfants, la consommation de cannabis n'entraîne pas de risque potentiel d'intoxication mortelle. Toutefois, le cannabis pourrait agir comme facteur déclenchant d'accident cardiovasculaire chez de rares individus prédisposés. Les individus jeunes, et/ou vulnérables ont un risque significativement plus élevé de développer une psychose à l'âge adulte ou de devenir dépendant au cannabis. Des études épidémiologiques ont montré que le risque de développer une schizophrénie à l'âge adulte était augmenté pour les consommateurs de cannabis et ceci d'autant plus que l'âge de début de consommation était précoce. Il en va de même pour le risque de dépression. Les troubles respiratoires pourraient être exacerbés par la prise de cannabis. Les femmes enceintes et celles qui allaitent ne devraient pas consommer de cannabis car le THC traverse la barrière hémato-placentaire, en outre, il se concentre dans le lait maternel. La période de la vie la plus sensible aux effets néfastes du cannabis correspond à celle allant du foetus à l'adolescent. Le système endocannabinoïde sur lequel agit le THC serait en effet un acteur majeur orchestrant le développement des réseaux neuronaux dans le cerveau immature. La prise concomitante d'autres psychotropes comme l'alcool, les benzodiazépines ou la cocaïne conduit à des renforcements mutuels de leurs effets délétères. De plus, il a été montré l'existence d'une sensibilité croisée pour la majorité des psychotropes qui agissent sur le système de la récompense, le cannabis y compris, ce qui augmente ainsi le risque de pharmacodépendance. La prise régulière de doses élevées de cannabis entraîne l'apparition d'une tolérance et de symptômes de sevrage discrets à l'arrêt de la consommation. À part les effets négatifs mentionnés auparavant, le cannabis possède des propriétés médicales originales qui sont l'objet d'études attentives. Plusieurs cannabinoïdes mineurs naturels ou synthétiques, comme l'acide ajulémique, pourraient trouver un jour une place dans la pharmacopée. En usage thérapeutique, des variétés particulières de cannabis sont préférées, par exemple celles riches en cannabidiol non psychoactif. Le mode d'administration diffère de celui utilisé en mode récréatif. Par exemple, la vaporisation des cannabinoïdes à basse température est préférée à l'inhalation du "joint".

Patterns of alcohol consumption in the Seychelles Islands (Indian Ocean).

Self-reported drinking habits were examined in a random sample of 1067 persons aged 25-64 years in the Seychelles, a country in epidemiological transition where consumption of home-brewed, mostly unregistered beverages has been traditionally high. Alcohol consumption was calculated from respondents reporting at least one drink per week ('regular drinkers'). Among men, 51.1% were regular drinkers and had average intake of 112.1 ml alcohol a day. Among women, 5.9% were regular drinkers and had 49.7 ml alcohol a day. Frequency of drinking, but not amount per drinker, was slightly less in the 25-34-year than older-age categories. Home-brews (mostly palm toddy and fermented sugar cane juice) were consumed by 52% of regular drinkers and accounted for 54% of the total alcohol intake reported by all regular drinkers. Based on the reported consumption by regular drinkers only, the average annual alcohol consumption amounted respectively to 20.7 litres and 1.2 litres per man and woman aged 25-64 years, or, using extrapolation, 13.2 litres and 0.8 litres per man and woman respectively of the total population. These values may underestimate the true figures by half, since reported beer consumption accounted for 53% of beer sales. Socio-economic status was associated strongly and inversely with home-brew consumption, but slightly and positively with consumption of commercially marketed beverages. Alcohol intake was associated with smoking, high-density lipoprotein cholesterol, carbohydrate-deficient transferrin and blood pressure, but not with age and body mass index. In conclusion, these data show high alcohol consumption in the Seychelles with an important gender difference, a large proportion of alcohol derived from home-brews, and opposite tendencies for the relationships between socio-economic status and home-made or commercially marketed beverages.

Alcohol drinking, the metabolic syndrome and diabetes in a population with high mean alcohol consumption.

AIMS: To investigate the relationship of alcohol consumption with the metabolic syndrome and diabetes in a population-based study with high mean alcohol consumption. Few data exist on these conditions in high-risk drinkers. METHODS: In 6172 adults aged 35-75 years, alcohol consumption was categorized as 0, 1-6, 7-13, 14-20, 21-27, 28-34 and ≥ 35 drinks/week or as non-drinkers (0), low-risk (1-13), medium-to-high-risk (14-34) and very-high-risk (≥ 35) drinkers. Alcohol consumption was objectively confirmed by biochemical tests. In multivariate analysis, we assessed the relationship of alcohol consumption with adjusted prevalence of the metabolic syndrome, diabetes and insulin resistance, determined with the homeostasis model assessment of insulin resistance (HOMA-IR). RESULTS: Seventy-three per cent of participants consumed alcohol, 16% were medium-to-high-risk drinkers and 2% very-high-risk drinkers. In multivariate analysis, the prevalence of the metabolic syndrome, diabetes and mean HOMA-IR decreased with low-risk drinking and increased with high-risk drinking. Adjusted prevalence of the metabolic syndrome was 24% in non-drinkers, 19% in low-risk (P<0.001 vs. non-drinkers), 20% in medium-to-high-risk and 29% in very-high-risk drinkers (P=0.005 vs. low-risk). Adjusted prevalence of diabetes was 6.0% in non-drinkers, 3.6% in low-risk (P<0.001 vs. non-drinkers), 3.8% in medium-to-high-risk and 6.7% in very-high-risk drinkers (P=0.046 vs. low-risk). Adjusted HOMA-IR was 2.47 in non-drinkers, 2.14 in low-risk (P<0.001 vs. non-drinkers), 2.27 in medium-to-high-risk and 2.53 in very-high-risk drinkers (P=0.04 vs. low-risk). These relationships did not differ according to beverage types. CONCLUSIONS: Alcohol has a U-shaped relationship with the metabolic syndrome, diabetes and HOMA-IR, without differences between beverage types.