KCTD Hetero-oligomers confer unique kinetic properties on Hippocampal GABA B Receptor-Induced K + Currents

GABAB receptors are the G-protein coupled receptors for the main inhibitory neurotransmitter in the brain, GABA. GABAB receptors were shown to associate with homo-oligomers of auxiliary KCTD8, KCTD12, KCTD12b, and KCTD16 subunits (named after their T1 K+-channel tetramerization domain) that regulate G-protein signaling of the receptor. Here we provide evidence that GABAB receptors also associate with hetero-oligomers of KCTD subunits. Coimmunoprecipitation experiments indicate that two-thirds of the KCTD16 proteins in the hippocampus of adult mice associate with KCTD12. We show that the KCTD proteins hetero-oligomerize through self-interacting T1 and H1 homology domains. Bioluminescence resonance energy transfer measurements in live cells reveal that KCTD12/KCTD16 hetero-oligomers associate with both the receptor and the G-protein. Electrophysiological experiments demonstrate that KCTD12/KCTD16 hetero-oligomers impart unique kinetic properties on G-protein-activated Kir3 currents. During prolonged receptor activation (one min) KCTD12/KCTD16 hetero-oligomers produce moderately desensitizing fast deactivating K+ currents, whereas KCTD12 and KCTD16 homo-oligomers produce strongly desensitizing fast deactivating currents and nondesensitizing slowly deactivating currents, respectively. During short activation (2 s) KCTD12/KCTD16 hetero-oligomers produce nondesensitizing slowly deactivating currents. Electrophysiological recordings from hippocampal neurons of KCTD knock-out mice are consistent with these findings and indicate that KCTD12/KCTD16 hetero-oligomers increase the duration of slow IPSCs. In summary, our data demonstrate that simultaneous assembly of distinct KCTDs at the receptor increases the molecular and functional repertoire of native GABAB receptors and modulates physiologically induced K+ current responses in the hippocampus

Determinants of smoking initiation among women in five European countries: a cross-sectional survey

<p>Abstract</p> <p>Background</p> <p>The rate of smoking and lung cancer among women is rising in Europe. The primary aim of this study was to determine why women begin smoking in five different European countries at different stages of the tobacco epidemic and to determine if smoking is associated with certain characteristics and/or beliefs about smoking.</p> <p>Methods</p> <p>A cross-sectional telephone survey on knowledge and beliefs about tobacco was conducted as part of the Women in Europe Against Lung Cancer and Smoking (WELAS) Project. A total of 5 000 adult women from France, Ireland, Italy, Czech Republic, and Sweden were interviewed, with 1 000 from each participating country. All participants were asked questions about demographics, knowledge and beliefs about smoking, and their tobacco use background. Current and former smokers also were asked questions about smoking initiation. Basic statistics on the cross-sectional data was reported with chi-squared and ANOVA p-values. Logistic regression was used to analyze ever versus never smokers. Linear regression analyses were used to analyze age of smoking initiation.</p> <p>Results</p> <p>Being older, being divorced, having friends/family who smoke, and having parents who smoke were all significantly associated with ever smoking, though the strength of the associations varied by country. The most frequently reported reason for initiation smoking was friend smoking, with 62.3% of ever smokers reporting friends as one of the reasons why they began smoking. Mean age of smoking initiation was 18.2 years and over 80% of participants started smoking by the age of 20. The highest levels of young initiators were in Sweden with 29.3% of women initiating smoking at age 14-15 and 12.0% initiating smoking younger than age 14. The lowest level of young initiators was in the Czech Republic with 13.7% of women initiating smoking at age 14-15 and 1.4% of women initiating smoking younger than age 14. Women who started smoking because their friends smoked or to look 'cool' were more likely to start smoking at a younger age. Women who started smoking to manage stress or to feel less depressed were more likely to start smoking at an older age.</p> <p>Conclusions</p> <p>In all five participating countries, friends were the primary factor influencing ever smoking, especially among younger women. The majority of participants began smoking in adolescence and the average reported age of smoking initiation was youngest in Sweden and oldest in the Czech Republic.</p

Cytisine for smoking cessation in patients with tuberculosis: a multicentre, randomised, double-blind, placebo-controlled phase 3 trial

Dogar O, Keding A, Gabe R, et al. Cytisine for smoking cessation in patients with tuberculosis: a multicentre, randomised, double-blind, placebo-controlled phase 3 trial. The Lancet. Global health. 2020;8(11):e1408-e1417.BACKGROUND: Smoking cessation is important in patients with tuberculosis because it can reduce the high rates of treatment failure and mortality. We aimed to assess the effectiveness and safety of cystine as a smoking cessation aid in patients with tuberculosis in Bangladesh and Pakistan.; METHODS: We did a randomised, double-blind, placebo-controlled, trial at 32 health centres in Bangladesh and Pakistan. Eligible patients were adults (aged >18 years in Bangladesh; aged >15 years in Pakistan) with pulmonary tuberculosis diagnosed in the previous 4 weeks, who smoked tobacco on a daily basis and were willing to stop smoking. Patients were randomly assigned (1:1) to receive behavioural support plus either oral cytisine (9 mg on day 0, which was gradually reduced to 1·5 mg by day 25) or placebo for 25 days. Randomisation was done using pregenerated block randomisation lists, stratified by trial sites. Investigators, clinicians, and patients were masked to treatment allocation. The primary outcome was continuous abstinence at 6 months, defined as self-report (of not having used more than five cigarettes, bidis, a water pipe, or smokeless tobacco products since the quit date), confirmed biochemically by a breath carbon monoxide reading of less than 10 parts per million. Primary and safety analysis were done in the intention-to-treat population. This trial is registered with the International Standard Randomised Clinical Trial Registry, ISRCTN43811467, and enrolment is complete.; FINDINGS: Between June 6, 2017, and April 30, 2018, 2472 patients (1527 patients from Bangladesh; 945 patients from Pakistan) were enrolled and randomly assigned to receive cytisine (n=1239) or placebo (n=1233). At 6 months, 401 (32·4%) participants in the cytisine group and 366 (29·7%) participants in the placebo group had achieved continuous abstinence (risk difference 2·68%, 95% CI -0·96 to 6·33; relative risk 1·09, 95% CI 0·97 to 1·23, p=0·114). 53 (4·3%) of 1239 participants in the cytisine group and 46 (3·7%) of 1233 participants in the placebo group reported serious adverse events (94 events in the cytisine group and 90 events in the placebo group), which included 91 deaths (49 in the cytisine group and 42 in the placebo group). None of the adverse events were attributed to the study medication.; INTERPRETATION: Our findings do not support the addition of cytisine to brief behavioural support for the treatment of tobacco dependence in patients with tuberculosis.; FUNDING: European Union Horizon 2020 and Health Data Research UK.; TRANSLATIONS: For the Bengali and Urdu translations of the abstract see Supplementary Materials section. Copyright © 2020 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license. Published by Elsevier Ltd.. All rights reserved

Nicotine delivery to users from cigarettes and from different types of e-cigarettes

BACKGROUND: Delivering nicotine in the way smokers seek is likely to be the key factor in e-cigarette (EC) success in replacing cigarettes. We examined to what degree different types of EC mimic nicotine intake from cigarettes. METHODS: Twelve participants (‘dual users’ of EC and cigarettes) used their own brand cigarette and nine different EC brands. Blood samples were taken at baseline and at 2-min intervals for 10 min and again at 30 min. RESULTS: Eleven smokers provided usable data. None of the EC matched cigarettes in nicotine delivery (C (max) = 17.9 ng/ml, T (max) = 4 min and AUC(0–>30) = 315 ng/ml/min). The EC with 48 mg/ml nicotine generated the closest PK profile (C (max) = 13.6 ng/ml, T (max) = 4 min, AUC(0–>30) = 245 ng/ml/min), followed by a third generation EC using 20 mg/ml nicotine (C (max) = 11.9 ng/ml, T (max) = 6 min, AUC(0–>30) = 232 ng/ml/min), followed by the tank system using 20 mg/ml nicotine (C (max) = 9.9 ng/ml, T (max) = 6 min, AUC(0–>30) = 201 ng/ml/min). Cig-a-like PK values were similar, ranging from C (max) 7.5 to 9.7 ng/ml, T (max) 4-6 min, and AUC(0–>30) 144 to 173 ng/ml/min. Moderate differences in e-liquid nicotine concentrations had little effect on nicotine delivery, e.g. the EC with 24 mg/ml cartridge had the same PK profile as ECs with 16 mg/ml cartridges. Using similar strength e-liquid, the tank EC provided significantly more nicotine than cig-a-like ECs. CONCLUSIONS: EC brands we tested do not deliver nicotine as efficiently as cigarettes, but newer EC products deliver nicotine more efficiently than cig-a-like brands. Moderate variations in nicotine content of e-liquid have little effect on nicotine delivery. Smokers who are finding cig-a-like EC unsatisfactory should be advised to try more advanced systems

Late Cretaceous-Cenozoic thermal evolution of the northern part of the Central Western Carpathians (Slovakia): revealed by zircon and apatite fission track thermochronology

Tectono-thermal evolution of the northern part of Central Western Carpathians (Slovakia) was revealed by zircon and apatite fission track thermochronology of Neogene deposits, volcanic rocks, and crystalline basement. New fission track ages combined with previous geochronological data are correlated with main tectonic events and important palaeogeographic changes. (1) The palaeo-Alpine burial to more than 10. km depths, heating (> 320 °C) and low-grade metamorphism of the crystalline basement was caused by crustal thickening due to nappe stacking driven by the collisional processes (~. 90-75. Ma). (2) Extensional collapse and exhumation of basement complexes during the Late Cretaceous to Middle Eocene (~. 75-40. Ma). (3) The extension process resulted in a new sedimentary cycle of the Central Carpathian Palaeogene Basin (~. 40-25. Ma) in the northern part of the Central Western Carpathians. During the ensuing burial under the thick sedimentary cover, the crystalline basement of the northern zone resided at 'hotter' conditions (ca > 120 °C and < 200 °C) which led to full annealing of apatite single-grain ages. In the meanwhile the southern zone of the crystalline basement was gradually exhumed to the depth of 5-3. km and occasionally appeared at the erosion surface. (4) Disintegration of the Central Carpathian Palaeogene Basin (~. 25-20. Ma) was closely connected with a partial exhumation of the Central Western Carpathians and intensive denudation. (5) The Middle to Late Miocene (~. 20-9. Ma) subduction of the Outer Carpathians substratum had crucial consequences for the tectonic evolution of the hinterland that was located in the upper crustal plate. The most external zone of the hinterland basement domain (Lúčanská and Krivánska Fatra Mts.) was exhumed to the 5-3. km depth and the intramontane basins were opened. (6) New fission track data and geomorphological criteria refer to the Pliocene-Quaternary mountain building processes in the external zones of the Central Western Carpathians. © 2014 Elsevier B.V

Attitudes of women from five European countries regarding tobacco control policies.

AimsTobacco-related cancers and, in particular, lung cancer still represents a substantial public health epidemic across Europe as a result of high rates of smoking prevalence. Countries in Europe have proposed and implemented tobacco control policies to reduce smoking prevalence, with some countries being more progressive than others. The aim of this study was to examine factors that influenced women's attitudes across five European countries relative to comprehensive smokefree laws in their countries.MethodsA cross-sectional landline telephone survey on attitudes towards tobacco control laws was conducted in five European countries: France, Ireland, Italy, the Czech Republic, and Sweden. Attitudinal scores were determined for each respondent relative to questions about smokefree laws. Logistic regression models were used to obtain odds ratios with 95% confidence intervals.ResultsA total of 5000 women were interviewed (1000 women from each country). The majority of women, regardless of smoking history, objected to smoking in public buses, enclosed shopping centers, hospitals, and other indoor work places. More women who had quit smoking believed that new tobacco control laws would prompt cessation - as compared with women who still smoked.ConclusionsIn general, there is very high support for national smokefree laws that cover bars, restaurants, and public transport systems. As such laws are implemented, attitudes do change, as demonstrated by the differences between countries such as Ireland and the Czech Republic. Implementing comprehensive smokefree laws will gain high approval and will be associated with prompting people to quit

TB and Tobacco trial

Dogar O, Barua D, Böckmann M, et al. The safety, effectiveness and cost-effectiveness of cytisine in achieving six-month continuous smoking abstinence in tuberculosis patients-protocol for a double-blind, placebo-controlled randomized trial. Addiction. 2018;113(9):1716-1726