Purification of Highly Active Alphavirus Replication Complexes Demonstrates Altered Fractionation of Multiple Cellular Membranes

Positive-strand RNA viruses replicate their genomes in membrane-associated structures; alphaviruses and many other groups induce membrane invaginations called spherules. Here, we established a protocol to purify these membranous replication complexes (RCs) from cells infected with Semliki Forest virus (SFV). We isolated SFV spherules located on the plasma membrane and further purified them using two consecutive density gradients. This revealed that SFV infection strongly modifies cellular membranes. We removed soluble proteins, the Golgi membranes, and most of the mitochondria, but plasma membrane, endoplasmic reticulum (ER), and late endosome markers were retained in the membrane fraction that contained viral RNA synthesizing activity, replicase proteins, and minus-and plus-strand RNA. Electron microscopy revealed that the purified membranes displayed spherule-like structures with a narrow neck. This membrane enrichment was specific to viral replication, as such a distribution of membrane markers was only observed after infection. Besides the plasma membrane, SFV infection remodeled the ER, and the cofractionation of the RC-carrying plasma membrane and ER suggests that SFV recruits ER proteins or membrane to the site of replication. The purified RCs were highly active in synthesizing both genomic and subgenomic RNA. Detergent solubilization destroyed the replication activity, demonstrating that the membrane association of the complex is essential. Most of the newly made RNA was in double-stranded replicative molecules, but the purified complexes also produced single-stranded RNA as well as released newly made RNA. This indicates that the purification established here maintained the functionality of RCs and thus enables further structural and functional studies of active RCs. IMPORTANCE Similar to all positive-strand RNA viruses, the arthropod-borne alpha-viruses induce membranous genome factories, but little is known about the arrangement of viral replicase proteins and the presence of host proteins in these replication complexes. To improve our knowledge of alphavirus RNA-synthesizing complexes, we isolated and purified them from infected mammalian cells. Detection of viral RNA and in vitro replication assays revealed that these complexes are abundant and highly active when located on the plasma membrane. After multiple purification steps, they remain functional in synthesizing and releasing viral RNA. Besides the plasma membrane, markers for the endoplasmic reticulum and late endosomes were enriched with the replication complexes, demonstrating that alphavirus infection modified cellular membranes beyond inducing replication spherules on the plasma membrane. We have developed here a gentle purification method to obtain large quantities of highly active replication complexes, and similar methods can be applied to other positive-strand RNA viruses.Peer reviewe

Depression-related work disability: socioeconomic inequalities in onset, duration and recurrence

Objective: Depression is a major cause of disability in working populations and the reduction of socioeconomic inequalities in disability is an important public health challenge. We examined work disability due to depression with four indicators of socioeconomic status. Methods: A prospective cohort study of 125 355 Finnish public sector employees was linked to national register data on work disability (>9 days) due to depressive disorders (International Classification of Diseases, codes F32–F34) from January 2005 to December 2011. Primary outcomes were the onset of work disability due to depressive disorders and, among those with such disability, return to work after and recurrent episodes of work disability due to depression. Results: We found a consistent inverse socioeconomic gradient in work disability due to depression. Lower occupational position, lower educational level, smaller residence size, and rented (vs. owner-occupied) residence were all associated with an increased risk of work disability. Return to work was slower for employees with basic education (cumulative odds ratio = 1.21, 95% CI: 1.05–1.39) compared to those with higher education. Recurrent work disability episodes due to depression were less common among upper-grade non-manual workers (the highest occupational group) than among lower-grade non-manual (hazard ratio = 1.16, 95% CI: 1.07–1.25) and manual (hazard ratio = 1.14, 95% CI: 1.02–1.26) workers. Conclusions: These data from Finnish public sector employees show persistent socioeconomic inequalities in work disability due to depression from 2005 to 2011 in terms of onset, recovery and recurrence

Predictors of employment in young adults with psychiatric work disability

AIM: Mental disorders are the leading cause of work disability among young adults in the industrialized world. Factors predicting employment after long-term psychiatric work disability are largely unknown. METHODS: We linked personal and clinical information from the benefit applications and medical certificates of 1163 young adults (18-34 years) with a new-onset fixed-term psychiatric disability pension in 2008 with employment records between 2005 and 2013. The outcomes were starting employment during and being employed at the end of follow-up. RESULTS: Of the participants, 48% had been employed during and 22% were employed at the end of follow-up. Sustained employment history, university education (master's degree) and no recorded psychological symptoms in childhood were associated with both subsequent employment outcomes. Women and participants under 25 years were more likely to start employment. Depression and other mental disorders (vs psychotic diagnose) and having no comorbid mental disorders or substance abuse were associated with employment at the end of follow-up. CONCLUSIONS: Sustained employment history, university education and no recorded psychological symptoms during childhood predict a return to employment among young adults after a fixed-term psychiatric work disability pension. Pro-active interventions in psychological problems during childhood could enhance employment after a period of work disability

Do predictors of return to work and recurrence of work disability due to mental disorders vary by age? A cohort study

BACKGROUND: The extent to which predictors of return to work (RTW) and recurrence of work disability episodes vary by age group is not well understood. METHODS: We examined the associations of sociodemographic and clinical factors with RTW and recurrence after mental-disorder-related work disability episodes in a cohort of 10,496 Finnish public sector employees. Disability records were derived from national disability registers between 2005 and 2011. Effect modification by age was examined in age groups of 21-34, 35-50 and >50 years. RESULTS: A total of 16,551 disability episodes from mental disorders were recorded. The likelihood of RTW was elevated in age group 21-34 (hazard ratio (HR) = 1.36, 95% confidence interval (CI) = 1.28-1.46) and 35-50 years (HR = 1.22, 95% CI = 1.18-1.26) compared to age group >50 years. The risk of a recurrent episode of work disability was higher in age groups >50 (HR = 1.29, 95% CI = 1.09-1.52) and 35-50 years (HR = 1.20, 95% CI = 1.03-1.41) compared to the youngest age group. Employees with depressive disorders were less likely to RTW than employees with neurotic, stress-related and somatoform disorders, and this difference increased with age. Low education was associated with increased risk of recurrent work disability episode in age groups of 50 years or younger, while no such association was observed in age group >50 years. CONCLUSIONS: The importance of depressive symptoms over neurotic, stress-related and somatoform disorders as predictors of delayed RTW increases with age, whereas educational differences in the recurrence of an episode diminish by age

Digitalisation in wind and solar power technologies

Highlights • Digitalisation and ICT solutions are impacting on wind power and solar PV technologies. • The prominent RES technologies with ICT solutions control, manage and optimise electricity production. • Wind power patent data shows a straightforward technology convergence trend with ICT. • Basic inventions in solar PV have increased more rapidly than solar PV ICT solutions. • Digitalisation in wind power and solar PV has been driven by the US, Germany, Denmark and Japan.Smart energy transition includes a widespread deployment of clean energy technologies and intelligent energy management with information and communication technologies (ICTs). In this paper, the smart energy transition is studied from the viewpoint of the technology convergence of renewable energy sources (RESs) and ICTs. Two important, fast-growing and weather-dependent renewable energy generation technologies: wind power and solar PV (photovoltaic) are studied. This paper provides technology convergence analyses of RES and ICT inventions based on international patent data. Digitalisation is changing the whole of society, and according to the results, this transition can also be seen in the studied renewable energy generation technologies. The digitalisation of RES production covers technologies that control, manage and optimise electricity production in different intelligent ways. Differences between wind power and solar PV technologies are found: in the case of wind power, the development from virtually no ICT solutions to partial technology convergence with the ICT sector is straightforward. However, in the case of solar PV, the development of basic technologies has been even faster than the development of the solar PV ICT solutions, which may indicate the immature nature of solar PV technologies during the studied years. The digitalisation of the renewable energy sector poses challenges for RES companies in following and predicting ICT development and opportunities for innovations and collaborations with ICT companies. This conclusion can also be expanded to society and policy levels because focusing on only a narrow field when planning innovation policy instruments can negatively impact the country's competitiveness

Weight Loss Trajectories in Healthy Weight Coaching : Cohort Study

Background: As global obesity prevalence continues to increase, there is a need for accessible and affordable weight management interventions, such as web-based programs. Objective: This paper aims to assess the outcomes of healthy weight coaching (HWC), a web-based obesity management program integrated into standard Finnish clinical care. Methods: HWC is an ongoing, structured digital 12-month program based on acceptance and commitment therapy. It includes weekly training sessions focused on lifestyle, general health, and psychological factors. Participants received remote one-on-one support from a personal coach. In this real-life, single-arm, prospective cohort study, we examined the total weight loss, weight loss profiles, and variables associated with weight loss success and program retention in 1189 adults (963 women) with a BMI >25 kg/m(2) among participants of the program between October 2016 and March 2019. Absolute (kg) and relative (%) weight loss from the baseline were the primary outcomes. We also examined the weight loss profiles, clustered based on the dynamic time-warping distance, and the possible variables associated with greater weight loss success and program retention. We compared different groups using the Mann-Whitney test or Kruskal-Wallis test for continuous variables and the chi-squared test for categorical variables. We analyzed changes in medication using the McNemar test. Results: Among those having reached the 12-month time point (n=173), the mean weight loss was 4.6% (SE 0.5%), with 43% (n=75) achieving clinically relevant weight loss (>= 5%). Baseline BMI >= 40 kg/m(2) was associated with a greater weight loss than a lower BMI (mean 6.6%, SE 0.9%, vs mean 3.2%, SE 0.6%; P=.02). In addition, more frequent weight reporting was associated with greater weight loss. No significant differences in weight loss were observed according to sex, age, baseline disease, or medication use. The total dropout rate was 29.1%. Dropouts were slightly younger than continuers (47.2, SE 0.6 years vs 49.2, SE 0.4 years; P=.01) and reported their weight less frequently (3.0, SE 0.1 entries per month vs 3.3, SE 0.1 entries per month; P Conclusions: A comprehensive web-based program such as HWC is a potential addition to the repertoire of obesity management in a clinical setting. Heavier patients lost more weight, but weight loss success was otherwise independent of baseline characteristics.Peer reviewe

Prognostic factors for return to work after depression-related work disability: A systematic review and meta-analysis.

Knowledge about factors influencing return to work (RTW) after depression-related absence is highly relevant, but the evidence is scattered. We performed a systematic search of PubMed and Embase databases up to February 1, 2016 to retrieve cohort studies on the association between various predictive factors and return to work among employees with depression for review and meta-analysis. We also analyzed unpublished data from the Finnish Public Sector study. Most-adjusted estimates were pooled using fixed effects meta-analysis. Eleven published studies fulfilled the eligibility criteria, representing 22 358 person-observations from five different countries. With the additional unpublished data from the 14 101 person-observations from the Finnish Public Sector study, the total number of person-observations was 36 459. The pooled estimates were derived from 2 to 5 studies, with the number of observations ranging from 260 to 26 348. Older age (pooled relative risk [RR] 0.95; 95% confidence interval [CI] 0.84-0.87), somatic comorbidity (RR = 0.80, 95% CI 0.77-0.83), psychiatric comorbidity (RR = 0.86, 95% CI 0.83-0.88) and more severe depression (RR = 0.96, 95% CI 0.94-0.98) were associated with a lower rate of return to work, and personality trait conscientiousness with higher (RR = 1.06, 95% CI 1.02-1.10) return to work. While older age and clinical factors predicted slower return, significant heterogeneity was observed between the studies. There is a dearth of observational studies on the predictors of RTW after depression. Future research should pay attention to quality aspects and particularly focus on the role of workplace and labor market factors as well as individual and clinical characteristics on RTW

CHARACTERISATION AND IN VITRO AND IN VIVO EVALUATION OF SUPERCRITICAL-CO2-FOAMED B-TCP/PLCL COMPOSITES FOR BONE APPLICATIONS

Most synthetic bone grafts are either hard and brittle ceramics or paste-like materials that differ in applicability from the gold standard autologous bone graft, which restricts their widespread use. Therefore, the aim of the study was to develop an elastic, highly porous and biodegradable beta-tricalciumphosphate/poly(L-lactide-co-epsilon-caprolactone) (beta-TCP/PLCL) composite for bone applications using supercritical CO2 foaming. Ability to support osteogenic differentiation was tested in human adipose stem cell (hASC) culture for 21 d. Biocompatibility was evaluated for 24 weeks in a rabbit femur-defect model. Foamed composites had a high ceramic content (50 wt%) and porosity (65-67 %). After 50 % compression, in an aqueous environment at 37 degrees C, tested samples returned to 95 % of their original height. Hydrolytic degradation of beta-TCP/PLCL composite, during the 24-week follow-up, was very similar to that of porous PLCL scaffold both in vitro and in vivo. Osteogenic differentiation of hASCs was demonstrated by alkaline phosphatase activity analysis, alizarin red staining, soluble collagen analysis, immunocytochemical staining and qRT-PCR. In vitro, hASCs formed a pronounced mineralised collagen matrix. A rabbit femur defect model confirmed biocompatibility of the composite. According to histological Masson-Goldner's trichrome staining and micro-computed tomography, beta-TCP/PLCL composite did not elicit infection, formation of fibrous capsule or cysts. Finally, native bone tissue at 4 weeks was already able to grow on and in the beta-TCP/PLCL composite. The elastic and highly porous beta-TCP/PLCL composite is a promising bone substitute because it is osteoconductive and easy-to-use and mould intraoperatively.Peer reviewe

Personal identity (de)formation among lifestyle travellers: A double-edged sword?

This article explores the personal identity work of lifestyle travellers – individuals for whom extended leisure travel is a preferred lifestyle that they return to repeatedly. Qualitative findings from in-depth semi-structured interviews with lifestyle travellers in northern India and southern Thailand are interpreted in light of theories on identity formation in late modernity that position identity as problematic. It is suggested that extended leisure travel can provide exposure to varied cultural praxes that may contribute to a sense of social saturation. Whilst a minority of the respondents embraced a saturation of personal identity in the subjective formation of a cosmopolitan cultural identity, several of the respondents were paradoxically left with more identity questions than answers as the result of their travels