Reduced neurosteroid potentiation of GABAA receptors in epilepsy and depolarized hippocampal neurons

OBJECTIVE: Neurosteroids regulate neuronal excitability by potentiating γ-aminobutyric acid type-A receptors (GABARs). In animal models of temporal lobe epilepsy, the neurosteroid sensitivity of GABARs is diminished and GABAR subunit composition is altered. We tested whether similar changes occur in patients with epilepsy and if depolarization-induced increases in neuronal activity can replicate this effect. METHODS: We determined GABAR α4 subunit expression in cortical tissue resected from pediatric epilepsy patients. Modulation of human GABARs by allopregnanolone and Ro15-4513 was measured in Xenopus oocytes using whole-cell patch clamp. To extend the findings obtained using tissue from epilepsy patients, we evaluated GABAR expression and modulation by allopregnanolone and Ro15-4513 in cultured rat hippocampal neurons exposed to high extracellular potassium (HK) to increase neuronal activity. RESULTS: Expression of α4 subunits was increased in pediatric cortical epilepsy specimens encompassing multiple pathologies. The potentiation of GABA-evoked currents by the neurosteroid allopregnanolone was decreased in Xenopus oocytes expressing GABARs isolated from epilepsy patients. Furthermore, receptors isolated from epilepsy but not control tissue were sensitive to potentiation by Ro15-4513, indicating higher expression of α INTERPRETATION: These findings suggest that seizure activity-induced upregulation of

Protocol: precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli.

Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are

An Extended Plant Circadian Clock Model for Characterising Flowering Time under Different Light Quality Conditions

Speed breeding has recently emerged as an innovative agricultural technology solution to meet the ever-increasing global food demand. In speed breeding, typically various light qualities (e.g., colour, duration, intensity) are modified to manipulate the circadian clock of the plants, which in turn alter the plant growth and enhance the productivity such as by reducing the flowering time. In order to develop a comprehensive framework describing plant growth, a model incorporating the effect of various light qualities on plant growth needs to be established. Recently a mathematical model of the plant circadian clock for Arabidopsis thaliana has been developed to characterise the hypocotyl growth subject to multiple light quality properties. This is a first step towards developing a more comprehensive model that links light quality, plant circadian clock and plant growth. In this work, we extend the model by adding the effect of various light qualities on the flowering time. The proposed model can capture the flowering time behaviours of plant when subject to red, blue, and mixed lights and can be used to guide experiment of light properties manipulation for optimised plant growth via hypocotyl growth and flowering time

Mathematical model of insulin kinetics accounting for the amino acids effect during a mixed meal tolerance test

Amino acids (AAs) are well known to be involved in the regulation of glucose metabolism and, in particular, of insulin secretion. However, the effects of different AAs on insulin release and kinetics have not been completely elucidated. The aim of this study was to propose a mathematical model that includes the effect of AAs on insulin kinetics during a mixed meal tolerance test. To this aim, five different models were proposed and compared. Validation was performed using average data, derived from the scientific literature, regarding subjects with normal glucose tolerance (CNT) and with type 2 diabetes (T2D). From the average data of the CNT and T2D people, data for two virtual populations (100 for each group) were generated for further model validation. Among the five proposed models, a simple model including one first-order differential equation showed the best results in terms of model performance (best compromise between model structure parsimony, estimated parameters plausibility, and data fit accuracy). With regard to the contribution of AAs to insulin appearance/disappearance (kAA model parameter), model analysis of the average data from the literature yielded 0.0247 (confidence interval, CI: 0.0168 - 0.0325) and -0.0048 (CI: -0.0281 - 0.0185) μU·ml-1/(μmol·l-1·min), for CNT and T2D, respectively. This suggests a positive effect of AAs on insulin secretion in CNT, and negligible effect in T2D. In conclusion, a simple model, including single first-order differential equation, may help to describe the possible AAs effects on insulin kinetics during a physiological metabolic test, and provide parameters that can be assessed in the single individuals

Longitudinal associations between self-reported sleep duration and cardiometabolic disease risk in corporate executives

Objective: This study aimed to determine the longitudinal associations between self-reported sleep duration and cardiometabolic disease (CMD) risk in corporate executives. Methods: Self-reported sleep duration and lifestyle, occupational, psychological, and anthropometrical, blood pressure and blood marker variables were obtained from 1512 employees at annual health risk assessments in South Africa between 2016 and 2019. Gender-stratified linear mixed models, adjusting for age, lifestyle, occupational and psychological covariates were used to explore these longitudinal associations. Results: Among women, shorter sleep duration was associated with higher body mass index (BMI) covarying for age only (ß with 95% confidence intervals: −0.19 [−0.36, −0.03]), age and occupational factors (−0.20 [−0.36, −0.03]) and age and psychological factors (−0.20 [−0.37, −0.03]). Among men, shorter sleep was associated with both BMI and waist circumference (WC) covarying for age only (BMI: −0.15 [−0.22; −0.08]; WC: −0.62 [−0.88; −0.37]); age and lifestyle factors (BMI: −0.12 [−0.21; −0.04]); WC: −0.016 [−0.92; −0.29], age and occupational factors (BMI: −0.20 [−0.22; 0.08]; WC: −0.62 [−0.88; −0.36]), and age and psychological factors (BMI: −0.15 [−0.22; −0.07]; WC: −0.59 [−0.86; −0.33]). Among men, shorter sleep was also longitudinally associated with higher CMD risk scores in models adjusted for age and lifestyle factors (CMD: −0.12 [−0.20; −0.04]) and age and psychological factors (CMD: −0.08 [−0.15; −0.01]). Conclusion: Corporate executives who report shorter sleep durations may present with poorer CMD risk profiles, independent of age, lifestyle, occupational and psychological factors. Addressing sleep health in workplace health programmes may help mitigate the development of CMD in such employees.</p

Association between self-reported sleep duration and cardiometabolic risk in corporate executives

Purpose: This cross-sectional study aimed to compare the association between self-reported sleep duration and cardiometabolic risk among men and women corporate executives and investigate potential lifestyle, work- and stress-related mediators thereof. Methods: Self-reported sleep duration and lifestyle, occupational, psychological and measured anthropometrical, blood pressure (BP) and blood marker variables were obtained from health risk assessment data of 3583 corporate executives. Sex-stratified regression analyses investigated the relationships between occupational and psychological variables with self-reported sleep duration, and sleep duration with individual cardiometabolic risk factors. Mediation analyses investigated the effects of work, psychological and lifestyle factors on the relationships between self-reported sleep duration and cardiometabolic risk factors, as well as a continuous cardiometabolic risk score calculated from the sum of sex-stratified z-standardized scores of negative fasting serum HDL, and positive plasma Glu, serum TG, body mass index (BMI), waist circumference, systolic and diastolic BP. Results: Longer work hours and work commute time, depression, anxiety and stress were associated with shorter sleep duration in both men and women (all p < 0.05). Shorter sleep duration was associated with higher BMI, larger waist circumference and greater cardiometabolic risk scores in both men and women (all p < 0.05), higher diastolic BP in men (p < 0.05) and lower HDL cholesterol in women (p < 0.05). Physical activity, working hours and stress significantly mediated the relationships between self-reported sleep duration and BMI, waist circumference, diastolic BP and cardiometabolic risk score in men only. Conclusion: In these corporate executives, shorter self-reported sleep duration is associated with poorer psychological, occupational and cardiometabolic risk outcomes in both men and women. Given that physical activity, working hours and stress mediate this association among the men, the case for sleep health interventions in workplace health programmes is warranted

Assessing the validity and reliability and determining cut-points of the Actiwatch 2 in measuring physical activity

Objective: The Actiwatch 2 (AW2) is a wrist-worn accelerometer typically used to measure sleep. Although it can measure physical activity, there is limited evidence supporting its validity. We assessed the validity and reliability of the AW2 to measure sedentary behavior and physical activity (light, moderate, vigorous intensities), and reported their respective count cut-points. Approach: Twenty-eight males and 22 females completed a task battery comprising three sedentary tasks and six randomized physical activity tasks at varying intensities, whilst wearing the AW2, a reference accelerometry device (Actigraph GT3X) and a cardiopulmonary gas analyzer on two separate occasions. Validity was assessed using correlations (AW2 counts versus GT3X counts and metabolic equivalent (MET) values), reliability using Bland–Altman analyses, and cut-points were determined using receiver operating characteristic (ROC) area under the curve (AUC) analyses. Main results: AW2 counts were positively correlated with GT3X counts (rho = 0.902, p < 0.001) and METs (rho = 0.900, p < 0.001). AW2-derived counts were comparable across independent assessment periods. Sedentary (AUC = 0.99, cut-point: 256 cpm) and vigorous activity (AUC = 0.95, cut-point: 720 cpm) were strongly characterized, and moderate activity (AUC = 0.66, cut-point: 418 cpm) was weakly characterized. Significance: The use of the AW2 in physical activity monitoring looks promising for sedentary behavior, moderate and vigorous activity, however, further validation is needed

Associations Between Self-Reported Sleep Duration and Mortality in Employed Individuals:Systematic Review and Meta-Analysis

Objective: Sleeping less or more than the 7-8 h has been associated with mortality in the general population, which encompasses diversity in employment status, age and community settings. Since sleep patterns of employed individuals may differ to those of their unemployed counterparts, the nature of their sleep-mortality relationship may vary. We therefore investigated the association between self-reported sleep duration and all-cause mortality (ACM) or cardiovascular disease mortality (CVDM) in employed individuals. Data sources: Based on Preferred Reporting Items for Systematic Reviews and Meta-Analyses, searches between January 1990 and May 2020 were conducted in PubMed, Web of Science and Scopus. Inclusion/exclusion criteria: Included were prospective cohort studies of 18–64-year-old disease-free employed persons with sleep duration measured at baseline, and cause of death recorded prospectively as the outcome. Gray literature, case-control or intervention design studies were excluded. Data Extraction: Characteristics of the studies, participants, and study outcomes were extracted. The quality and risk of bias were assessed using the Newcastle-Ottawa Scale. Data synthesis: The pooled relative risks (RR) with 95% confidence intervals (CI) were obtained with a random-effects model and results presented as forest plots. Heterogeneity and sensitivity analysis were assessed. Results: Shorter sleep duration (less than or equal to 6 h) was associated with a higher risk for (ACM) (RR: 1.16, 95% CI: 1.11 -1.22) and CVDM (RR: 1.26, 95% CI: 1.12 -1.41) compared to 7-8 h of sleep, with no significant heterogeneity. The association between longer sleep (greater than or equal to 8 h) and ACM (RR: 1.18, 95% CI:1.12 -1.23, P < 0.001) needs to be interpreted cautiously owing to high heterogeneity (I2 ¼ 86.0%, P < 0.001). Conclusion: Interventions and education programs targeting sleep health in the workplace may be warranted, based on our findings that employed individuals who report shorter sleep appear to have a higher risk for ACM and CVDM