Programmable synthetic biomolecular condensates for cellular control.

The formation of biomolecular condensates mediated by a coupling of associative and segregative phase transitions plays a critical role in controlling diverse cellular functions in nature. This has inspired the use of phase transitions to design synthetic systems. While design rules of phase transitions have been established for many synthetic intrinsically disordered proteins, most efforts have focused on investigating their phase behaviors in a test tube. Here, we present a rational engineering approach to program the formation and physical properties of synthetic condensates to achieve intended cellular functions. We demonstrate this approach through targeted plasmid sequestration and transcription regulation in bacteria and modulation of a protein circuit in mammalian cells. Our approach lays the foundation for engineering designer condensates for synthetic biology applications.

Chemical Aspects of Human and Environmental Overload with Fluorine.

Over the last 100-120 years, due to the ever-increasing importance of fluorine-containing compounds in modern technology and daily life, the explosive development of the fluorochemical industry led to an enormous increase of emission of fluoride ions into the biosphere. This made it more and more important to understand the biological activities, metabolism, degradation, and possible environmental hazards of such substances. This comprehensive and critical review focuses on the effects of fluoride ions and organofluorine compounds (mainly pharmaceuticals and agrochemicals) on human health and the environment. To give a better overview, various connected topics are also discussed: reasons and trends of the advance of fluorine-containing pharmaceuticals and agrochemicals, metabolism of fluorinated drugs, withdrawn fluorinated drugs, natural sources of organic and inorganic fluorine compounds in the environment (including the biosphere), sources of fluoride intake, and finally biomarkers of fluoride exposure.

Fistula development after anal abscess drainage-a multicentre retrospective cohort study.

PURPOSE: Anal abscesses are common and, despite correct treatment with surgical drainage, carry the risk of developing fistulas. Studies identifying risk factors for the development of anal fistulas are sparse. This study aimed to identify the risk factors for anal fistulas after anal abscess surgery. METHODS: This was a multicentre, retrospective cohort study of patients undergoing acute surgery for anal abscesses in the Capital Region of Denmark between 2018 and 2019. The patients were identified using ICD-10 codes for anal abscesses. Predefined clinicopathological factors and postoperative courses were extracted from patient records. RESULTS: A total of 475 patients were included. At a median follow-up time of 1108 days (IQR 946-1320 days) following surgery, 164 (33.7%) patients were diagnosed with an anal fistula. Risk factors for developing fistulas were low intersphincteric (OR 2.77, 95CI 1.50-5.06) and ischioanal (OR 2.48, 95CI 1.36-4.47) abscesses, Crohn's disease (OR 5.96, 95CI 2.33-17.2), a history of recurrent anal abscesses (OR 4.14, 95CI 2.47-7.01) or repeat surgery (OR 5.96, 95CI 2.33-17.2), E. coli-positive pus cultures (OR 4.06, 1.56-11.4) or preoperative C-reactive protein (CRP) of more than 100 mg/L (OR 3.21, 95CI 1.57-6.71). CONCLUSION: Several significant clinical risk factors were associated with fistula development following anal abscess surgery. These findings are clinically relevant and could influence the selection of patients for specialised follow-up, facilitate expedited diagnosis, and potentially prevent unnecessarily long treatment courses.

Particle Size Modulates Silver Nanoparticle Toxicity during Embryogenesis of Urchins Arbacia lixula and Paracentrotus lividus.

Silver nanoparticles represent a threat to biota and have been shown to cause harm through a number of mechanisms, using a wide range of bioassay endpoints. While nanoparticle concentration has been primarily considered, comparison of studies that have used differently sized nanoparticles indicate that nanoparticle diameter may be an important factor that impacts negative outcomes. In considering this, the aim of the present study was to determine if different sizes of silver nanoparticles (AgNPs; 10, 20, 40, 60 and 100 nm) give rise to similar effects during embryogenesis of Mediterranean sea urchins Arbacia lixula and Paracentrotus lividus, or if nanoparticle size is a parameter that can modulate embryotoxicity and spermiotoxicity in these species. Fertilised embryos were exposed to a range of AgNP concentrations (1−1000 µg L−1) and after 48 h larvae were scored. Embryos exposed to 1 and 10 µg L−1 AgNPs (for all tested sizes) showed no negative effect in both sea urchins. The smaller AgNPs (size 10 and 20 nm) caused a decrease in the percentage of normally developed A. lixula larvae at concentrations ≥50 µg L−1 (EC50: 49 and 75 µg L−1, respectively) and at ≥100 µg L−1 (EC50: 67 and 91 µg L−1, respectively) for P. lividus. AgNPs of 40 nm diameter was less harmful in both species ((EC50: 322 and 486 µg L−1, for P. lividus and A. lixula, respectively)). The largest AgNPs (60 and 100 nm) showed a dose-dependent response, with little effect at lower concentrations, while more than 50% of larvae were developmentally delayed at the highest tested concentrations of 500 and 1000 µg L−1 (EC50(100 nm); 662 and 529 µg L−1, for P. lividus and A. lixula, respectively. While AgNPs showed no effect on the fertilisation success of treated sperm, an increase in offspring developmental defects and arrested development was observed in A. lixula larvae for 10 nm AgNPs at concentrations ≥50 µg L−1, and for 20 and 40 nm AgNPs at concentrations >100 µg L−1. Overall, toxicity was mostly ascribed to more rapid oxidative dissolution of smaller nanoparticles, although, in cases, Ag+ ion concentrations alone could not explain high toxicity, indicating a nanoparticle-size effect.

Ribosylation of the CD8αß heterodimer permits binding of the nonclassical major histocompatibility molecule, H2-Q10.

The CD8αß heterodimer plays a crucial role in the stabilization between major histocompatibility complex class I molecules (MHC-I) and the T cell receptor (TCR). The interaction between CD8 and MHC-I can be regulated by posttranslational modifications, which are proposed to play an important role in the development of CD8 T cells. One modification that has been proposed to control CD8 coreceptor function is ribosylation. Utilizing NAD+, the ecto-enzyme adenosine diphosphate (ADP) ribosyl transferase 2.2 (ART2.2) catalyzes the addition of ADP-ribosyl groups onto arginine residues of CD8α or ß chains and alters the interaction between the MHC and TCR complexes. To date, only interactions between modified CD8 and classical MHC-I (MHC-Ia), have been investigated and the interaction with non-classical MHC (MHC-Ib) has not been explored. Here, we show that ADP-ribosylation of CD8 facilitates the binding of the liver-restricted nonclassical MHC, H2-Q10, independent of the associated TCR or presented peptide, and propose that this highly regulated binding imposes an additional inhibitory leash on the activation of CD8-expressing cells in the presence of NAD+. These findings highlight additional important roles for nonclassical MHC-I in the regulation of immune responses.

Operationalising the 20-minute neighbourhood.

BACKGROUND: Recent rapid growth in urban areas and the desire to create liveable neighbourhoods has brought about a renewed interest in planning for compact cities, with concepts like the 20-minute neighbourhood (20MN) becoming more popular. A 20MN broadly reflects a neighbourhood that allows residents to meet their daily (non-work) needs within a short, non-motorised, trip from home. The 20MN concept underpins the key planning strategy of Australia's second largest city, Melbourne, however the 20MN definition has not been operationalised. This study aimed to develop and operationalise a practical definition of the 20MN and apply this to two Australian state capital cities: Melbourne (Victoria) and Adelaide (South Australia). METHODS: Using the metropolitan boundaries for Melbourne and Adelaide, data were sourced for several layers related to five domains: 1) healthy food; 2) recreational resources; 3) community resources; 4) public open space; and 5) public transport. The number of layers and the access measures required for each domain differed. For example, the recreational resources domain only required a sport and fitness centre (gym) within a 1.5-km network path distance, whereas the public open space domain required a public open space within a 400-m distance along a pedestrian network and 8 ha of public open space area within a 1-km radius. Locations that met the access requirements for each of the five domains were defined as 20MNs. RESULTS: In Melbourne 5.5% and in Adelaide 7.6% of the population were considered to reside in a 20MN. Within areas classified as residential, the median number of people per square kilometre with a 20MN in Melbourne was 6429 and the median number of dwellings per square kilometre was 3211. In Adelaide's 20MNs, both population density (3062) and dwelling density (1440) were lower than in Melbourne. CONCLUSIONS: The challenge of operationalising a practical definition of the 20MN has been addressed by this study and applied to two Australian cities. The approach can be adapted to other contexts as a first step to assessing the presence of existing 20MNs and monitoring further implementation of this concept.

Anthropometric cut-points for discriminating diabetes and the metabolic syndrome among Arabs and Asians: the Kuwait Diabetes Epidemiology Program.

This study aimed to determine anthropometric cut-points for screening diabetes and the metabolic syndrome (MetS) in Arab and South Asian ethnic groups in Kuwait and to compare the prevalence of the MetS based on the ethnic-specific waist circumference (WC) cut-point and the International Diabetes Federation (IDF) and American Heart Association/National Heart, Lung, and Blood Institute WC criteria. The national population-based survey data set of diabetes and obesity in Kuwait adults aged 18-60 years was analysed. Age-adjusted logistic regression and receiver operating characteristic (ROC) analyses were conducted to evaluate for 3589 individuals the utility of WC, waist:height ratio (WHtR) and BMI to discriminate both diabetes and ≥3 CVD risk factors. Areas under the ROC curve were similar for WC, WHtR and BMI. In Arab men, WC, WHtR and BMI cut-offs for diabetes were 106 cm, 0·55 and 28 kg/m2 and for ≥3 CVD risk factors, 97 cm, 0·55 and 28 kg/m2, respectively. In Arab women, cut-offs for diabetes were 107 cm, 0·65 and 33 kg/m2 and for ≥3 CVD risk factors, 93 cm, 0·60 and 30 kg/m2, respectively. WC cut-offs were higher for South Asian women than men. IDF-based WC cut-offs corresponded to a higher prevalence of the MetS across sex and ethnic groups, compared with Kuwait-specific cut-offs. Any of the assessed anthropometric indices can be used in screening of diabetes and ≥3 CVD risk factors in Kuwaiti Arab and Asian populations. ROC values were similar. The WC threshold for screening the MetS in Kuwaiti Arabs and South Asians is higher for women.

Uncommon side effects of common drugs in patients with familial dysautonomia.

PURPOSE: Patients with the autosomal recessive disorder of familial dysautonomia typically exhibit exacerbated adverse side effects to many common drugs. We aimed to catalog these adverse effects - with a focus on common drugs that are frequently administered to FD patients and compare their incidences to those within the general population. METHODS: We used data of 595 FD patients from an international database with information on drugs received and adverse effects. To investigate the molecular causes of reported differences in drug responses in FD patients, we used expression microarrays to compare the mRNA expression profiles in peripheral blood leukocytes of FD patients (n = 12) and healthy individuals (n = 10). RESULTS: Several drug classes, including cholinergics, anti-cholinergics, anti-convulsants, methylxanthines, SSRIs, and antibiotics caused either unreported symptoms or elevated rates of adverse events in FD patients. FD patients experienced different or more frequent adverse side effects than the general population in 31/123 drugs. These side effects included blood cell dyscrasias, amenorrhea, gastrointestinal bleeding, and bronchospasm. New findings include enhanced reaction of FD patients to H2 antagonist agents and to serotonin receptor agonists. We also detected eight genes differentially expressed between FD patients and healthy individuals that may underlie the differential drug responses of FD patients. CONCLUSION: We provide evidence that suggests the use of several common drugs should be discontinued or reduced in FD patients.

Team IHMC at the 2020 Cybathlon: a user-centered approach towards personal mobility exoskeletons.

BACKGROUND: The past few decades have seen rapid advancements in exoskeleton technology, with a considerable shift towards applications involving users with gait pathologies. Commercial devices from ReWalk, Ekso Bionics, and Indego, mainly designed for rehabilitation purposes, have inspired the development of many research platforms aimed at extending capabilities for use as safe and effective personal mobility devices. The 2016 Cybathlon featured an impressive demonstration of exoskeletons designed to enable mobility for individuals with spinal cord injury, however, not a single team completed every task and only two completed the stairs. Major improvements were showcased at the 2020 Cybathlon, with seven of the nine teams completing a similar set of tasks. Team IHMC built upon its silver-medal success from 2016 with an upgraded device, Quix. METHODS: Quix features several notable improvements including an additional powered degree of freedom for hip ab/adduction to laterally shift the device and reduce user effort while walking, custom-tailored cuffs and soft goods based on 3D body scans to optimize user comfort, and a streamlined testing pipeline for online tuning of gait parameters. RESULTS: Team IHMC finished in fourth place behind the teams from EPFL and Angel Robotics. Although we suffered from a considerably slower flat-ground walking speed, our pilot reported marked improvements in overall effort, comfort, and ease-of-use compared to our previous device. CONCLUSIONS: Clear progress in exoskeleton development has been exhibited since the inaugural Cybathlon, with tasks involving rough terrain, stairs, and ramps now posing little threat to most of the competitors. As a result, the layout of the powered exoskeleton course will likely undergo significant modifications to further push the devices towards suitability for personal everyday use. The current tasks do not address the issue of donning and doffing, nor do they simulate a scenario similar to maneuvering a kitchen to prepare a meal, for example. An additional limitation that may be more difficult to test in a competition setting is the required upper-body effort to manipulate the device in an effective manner.

The conflict between Public Health and 'Big Food': A Counterinsurgency?

This commentary characterises as an insurgency those elements of the Food Industry seeking to retain and expand industry profit-making opportunity with scant regard to the effect on public health and wellbeing. Premised on a conflict in which the battlespace encompasses the drivers of (un)healthful behaviour, our view encourages opponents of Food Industry methods to consider an approach based on analogy of successful Counter-Insurgency strategies to a health promotion context.

The murine CD94/NKG2 ligand, Qa-1b, is a high-affinity, functional ligand for the CD8αα homodimer.

The immune co-receptor CD8 molecule (CD8) has two subunits, CD8α and CD8ß, which can assemble into homo or heterodimers. Nonclassical (class-Ib) major histocompatibility complex (MHC) molecules (MHC-Ibs) have recently been identified as ligands for the CD8αα homodimer. This was demonstrated by the observation that histocompatibility 2, Q region locus 10 (H2-Q10) is a high-affinity ligand for CD8αα which also binds the MHC-Ib molecule H2-TL. This suggests that MHC-Ib proteins may be an extended source of CD8αα ligands. Expression of H2-T3/TL and H2-Q10 is restricted to the small intestine and liver, respectively, yet CD8αα-containing lymphocytes are present more broadly. Therefore, here we sought to determine whether murine CD8αα binds only to tissue-specific MHC-Ib molecules or also to ubiquitously expressed MHC-Ib molecules. Using recombinant proteins and surface plasmon resonance-based binding assays, we show that the MHC-Ib family furnishes multiple binding partners for murine CD8αα, including H2-T22 and the CD94/NKG2-A/B-activating NK receptor (NKG2) ligand Qa-1b We also demonstrate a hierarchy among MHC-Ib proteins with respect to CD8αα binding, in which Qa-1b > H2-Q10 > TL. Finally, we provide evidence that Qa-1b is a functional ligand for CD8αα, distinguishing it from its human homologue MHC class I antigen E (HLA-E). These findings provide additional clues as to how CD8αα-expressing cells are controlled in different tissues. They also highlight an unexpected immunological divergence of Qa-1b/HLA-E function, indicating the need for more robust studies of murine MHC-Ib proteins as models for human disease.

Associations between breast cancer screening participation and residential area sociodemographic features, geographic accessibility, and features of screening venue location in Greater Sydney, Australia.

Breast cancer screening (BCS) participation rates are often suboptimal and vary geographically. Environmental features may influence BCS participation, but few studies have assessed this relationship. This study assessed the associations between BCS participation, residential area sociodemographic characteristics, distance to BCS venue, and venue location attributes. Data for 384,433 women residing in Greater Sydney, Australia, invited to BCS during 2011-2014 were spatially joined to their state suburb (SSC) (n = 800). SSC sociodemographic measures included women's median age, proportion women speaking English at home, full-time employed, and university educated; and proportion dwellings with motor-vehicles. Road network distance was calculated to each BCS venue. BCS venues were coded as co-located with bus-stop, train-station, hospital, general practitioner (GP), and shop. Hot spots were calculated to quantify spatial clustering of BCS participation. Multilevel logistic models were used to estimate the associations between environmental predictors and BCS participation, accounting for SSC-level clustering. BCS participation was 53.9% and spatially clustered. BCS was positively associated with SSC-level median age for women, proportions women speaking English and university educated, and dwellings with motor-vehicles. Distance to venue was inversely associated with BCS. Venue co-location with GP was positively associated and co-location with bus-stop, train-station, and shop, hospital were negatively associated with BCS. Residential sociodemographic features, geographic access, and venue location attributes are associated with BCS participation. These findings implicate the relevance of social and built environmental factors to programmatic aims to raise BCS participation. Additional research on venue location features is required to understand where best to site BCS venues.

Associations between supermarket availability and body size in Australia: a cross-sectional observational study comparing state and territory capital cities.

BACKGROUND: Residential environment features such as availability of supermarkets may shape dietary behaviour and thus overweight and obesity. This relationship may not be consistent between cities. This Australian national-level study examined: 1) the relationship between supermarket availability and body size; and 2) whether this relationship varied by capital city. METHODS: This study used 2017-18 Australian National Health Survey data including individual-level socio-demographic information (age, sex, country of birth, education, occupation, household income), and measured body size (height and weight to derive body mass index [BMI], and waist circumference [WC]). Objectively-expressed measures of residential environments included: counts of supermarkets (major chain outlets), counts of amenities (representing walkable destinations including essential services, recreation, and entertainment), and area of public open space - each expressed within road-network buffers at 1000 m and 1500 m; population density (1km2 grid cells); and neighbourhood disadvantage (Index of Relative Socioeconomic Disadvantage) expressed within Statistical Area Level 1 units. Data for adult respondents ≥18 years residing in each of Australia's state and territory capital cities (n = 9649) were used in multilevel models to estimate associations between supermarket availability and body size sequentially accounting for individual and other environment measures. An interaction term estimated city-specific differences in associations between supermarket availability and body size. Models were consequently repeated stratified by city. RESULTS: Body size (BMI and WC) and supermarket availability varied between cities. Initial inverse associations between supermarket availability and body size (BMI and WC) were attenuated to null with inclusion of all covariates, except for BMI in the 1000 m buffer model (beta = - 0.148, 95%CI -0.27, - 0.01, p = 0.025). In stratified analyses, the strengths of associations varied between cities, remaining statistically significant only for some cities (BMI: Melbourne, Brisbane Hobart; WC: Brisbane, Hobart) in fully adjusted models. Different patterns of attenuation of associations with inclusion of covariates were evident for different cities. CONCLUSIONS: For Australian capital cities, greater availability of supermarkets is associated with healthful body size. Marked between-city variations in body size, supermarket availability, and relationships between supermarket availability and body size do not, however, support universal, "one-size-fits-all" solutions to change built environments to support healthful body size.

Prevalence of overweight and obesity, and associations with socio-demographic factors in Kuwait.

BACKGROUND: Kuwait is amongst countries in the Gulf region with high income economy. According to the World Health Organisation (WHO), one in five adults in the Gulf region is obese. This study sought to evaluate the prevalence and magnitude of association between overweight, obesity, central obesity, and socio-demographic factors in Kuwait. METHODS: A population-based cross-sectional survey of diabetes and obesity in Kuwait - part of the Kuwait Diabetes Epidemiology Program - was conducted between 2011 and 2014, targeting adults aged 18-82 years using the WHO STEPwise approach to non-communicable disease surveillance. Body mass index (BMI) was calculated to classify overweight and obesity, and waist circumference (WC) used to express central obesity. Multivariable logistic regression was used to estimate relationships between socio-demographic factors, overweight (25.0-29.9 kg/m2), obesity (≥30.0 kg/m2) or central obesity (WC ≥ 80 cm women; WC ≥ 94 cm men). RESULTS: Records for gender (56% Men), age, BMI, governorate, and nationality existed for 4901 individuals. Mean age and BMI were 43 years and 30 kg/m2, respectively. Non-Kuwaiti nationals were more prevalent than Kuwaitis (76% vs 24%). Prevalence rates for overweight, obesity and central obesity were 40.6% (95%CI: 38.4-42.8%), 42.1% (95%CI: 40.0-44.3%) and 73.7% (95%CI: 71.7-75.6%), respectively. The youngest age group (18-29 years) had rates of 38.2% (95%CI: 29.2-47.7%), 27.2% (95%CI: 19.0-36.7%) and 49.9% (95%CI: 40.6-59.1%) for overweight, obesity and central obesity, respectively. In covariate-adjusted analyses, the odds of being overweight was 26% greater for men than for women. Conversely, women had a 54% (95%CI: 19-99%) and 7-fold (95%CI, 5-10-fold) greater odds of obesity/central obesity, respectively, than men. Greater educational attainment, physical activity, and non-Kuwaiti status were associated with lower odds of obesity/central obesity. History of smoking, elevated blood pressure, higher income, being married, greater age and female sex related to greater odds of obesity/central obesity. CONCLUSION: Overweight was greater in men, obesity greater in women. Overweight and obesity prevalence were high in young adults aged 18-29 years, a significant public health concern. Efforts to integrate mandatory physical education to the school curriculum and promoting the creation of recreation spaces/parks to promote physical activities, will play a vital role in the early prevention of overweight/obesity in Kuwait.

Methods for accounting for neighbourhood self-selection in physical activity and dietary behaviour research: a systematic review.

BACKGROUND: Self-selection into residential neighbourhoods is a widely acknowledged, but under-studied problem in research investigating neighbourhood influences on physical activity and diet. Failure to handle neighbourhood self-selection can lead to biased estimates of the association between the neighbourhood environment and behaviour. This means that effects could be over- or under-estimated, both of which have implications for public health policies related to neighbourhood (re)design. Therefore, it is important that methods to deal with neighbourhood self-selection are identified and reviewed. The aim of this review was to assess how neighbourhood self-selection is conceived and accounted for in the literature. METHODS: Articles from a systematic search undertaken in 2017 were included if they examined associations between neighbourhood environment exposures and adult physical activity or dietary behaviour. Exposures could include any objective measurement of the built (e.g., supermarkets), natural (e.g., parks) or social (e.g., crime) environment. Articles had to explicitly state that a given method was used to account for neighbourhood self-selection. The systematic review was registered with the PROSPERO International Prospective Register of Systematic Reviews (number CRD42018083593) and was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. RESULTS: Of 31 eligible articles, almost all considered physical activity (30/31); few examined diet (2/31). Methods used to address neighbourhood self-selection varied. Most studies (23/31) accounted for items relating to participants' neighbourhood preferences or reasons for moving to the neighbourhood using multi-variable adjustment in regression models (20/23) or propensity scores (3/23). Of 11 longitudinal studies, three controlled for neighbourhood self-selection as an unmeasured confounder using fixed effects regression. CONCLUSIONS: Most studies accounted for neighbourhood self-selection by adjusting for measured attributes of neighbourhood preference. However, commonly the impact of adjustment could not be assessed. Future studies using adjustment should provide estimates of associations with and without adjustment for self-selection; consider temporality in the measurement of self-selection variables relative to the timing of the environmental exposure and outcome behaviours; and consider the theoretical plausibility of presumed pathways in cross-sectional research where causal direction is impossible to establish.

Coating-Dependent Effects of Silver Nanoparticles on Tobacco Seed Germination and Early Growth.

Silver nanoparticles (AgNPs) are used in a wide range of consumer products because of their excellent antimicrobial properties. AgNPs released into the environment are prone to transformations such as aggregation, oxidation, or dissolution so they are often stabilised by coatings that affect their physico-chemical properties and change their effect on living organisms. In this study we investigated the stability of polyvinylpyrrolidone (PVP) and cetyltrimethylammonium bromide (CTAB) coated AgNPs in an exposure medium, as well as their effect on tobacco germination and early growth. AgNP-CTAB was found to be more stable in the solid Murashige and Skoog (MS) medium compared to AgNP-PVP. The uptake and accumulation of silver in seedlings was equally efficient after exposure to both types of AgNPs. However, AgNP-PVP induced only mild toxicity on seedlings growth, while AgNP-CTAB caused severe negative effects on all parameters, even compared to AgNO3. Moreover, CTAB coating itself exerted negative effects on growth. Cysteine addition generally alleviated AgNP-PVP-induced negative effects, while it failed to improve germination and growth parameters after exposure to AgNP-CTAB. These results suggest that the toxic effects of AgNP-PVP are mainly a consequence of release of Ag+ ions, while phytotoxicity of AgNP-CTAB can rather be ascribed to surface coating itself.

Testosterone therapy to prevent type 2 diabetes mellitus in at-risk men (T4DM): Design and implementation of a double-blind randomized controlled trial.

BACKGROUND: Low circulating testosterone is associated with an increased risk of developing type 2 diabetes (T2DM) in overweight men with impaired glucose tolerance (IGT). AIMS: To determine in a multi-centre, double-blinded placebo-controlled randomized trial whether testosterone treatment combined with lifestyle intervention (Weight Watchers) relative to lifestyle intervention alone reduces T2DM incidence and improves glucose tolerance at 2 years. STUDY POPULATION: Overweight or obese men aged 50-74 years with a serum testosterone of ≤14 nmol/L and IGT or newly diagnosed T2DM established by an oral glucose tolerance test (OGTT). SETTING, DRUG AND PROTOCOL: Six Australian capital city-based tertiary care centres. Participants were randomized 1:1 and injected with testosterone undecanoate (1000 mg/4 mL) or vehicle (4 mL castor oil), at baseline, 6 weeks and 3-monthly thereafter. PRIMARY ENDPOINTS: (a) Proportion of participants with 2-hour OGTT ≥11.1 mmol/L at 2 years, and (b) a difference at 2 years ≥0.6 mmol/L in the mean 2-hour OGTT glucose between treatments. SECONDARY ENDPOINTS: Fasting insulin, HbA1c, body composition, maximal handgrip strength; sexual function and lower urinary tract symptoms; serum sex steroids and sex hormone binding globulin; mood and psychosocial function; adherence to lifestyle intervention; and healthcare utilization and costs. SAFETY: Overseen by an Independent Data Safety Monitoring Committee. Haematocrit, lipids and prostate-specific antigen (PSA) are assessed 6-monthly and information relating to haematological, urological and cardiovascular adverse events from each clinic visit. SUB-STUDIES: (a) Changes in bone density and micro-architecture, (b) motivation and behaviour, (c) telomere length, (d) extended treatment up to 4 years, and (e) hypothalamo-pituitary testicular axis recovery at treatment end.

A high-volume, low-cost approach to participant screening and enrolment: Experiences from the T4DM diabetes prevention trial.

BACKGROUND/AIMS: Participant recruitment to diabetes prevention randomised controlled trials is challenging and expensive. The T4DM study, a multicentre, Australia-based, Phase IIIb randomised controlled trial of testosterone to prevent Type 2 diabetes in men aged 50-74 years, faced the challenge of screening a large number of prospective participants at a small number of sites, with few staff, and a limited budget for screening activities. This article evaluates a high-volume, low-cost, semi-automated approach to screen and enrol T4DM study participants. METHODS: We developed a sequential multi-step screening process: (1) web-based pre-screening, (2) laboratory screening through a network of third-party pathology centres, and (3) final on-site screening, using online data collection, computer-driven eligibility checking, and automated, email-based communication with prospective participants. Phone- and mail-based data collection and communication options were available to participants at their request. The screening process was administered by the central coordinating centre through a central data management system. RESULTS: Screening activities required staffing of approximately 1.6 full-time equivalents over 4 years. Of 19,022 participants pre-screened, 13,108 attended a third-party pathology collection centre for laboratory screening, 1217 received final, on-site screening, and 1007 were randomised. In total, 95% of the participants opted for online pre-screening over phone-based pre-screening. Screening costs, including both direct and staffing costs, totalled AUD1,420,909 (AUD75 per subject screened and AUD1411 per randomised participant). CONCLUSION: A multi-step, semi-automated screening process with web-based pre-screening facilitated low-cost, high-volume participant enrolment to this large, multicentre randomised controlled trial. Centralisation and automation of screening activities resulted in substantial savings compared to previous, similar studies. Our screening approach could be adapted to other randomised controlled trial settings to minimise the cost of screening large numbers of participants.

Rapid Production of Virus Protein Microarray Using Protein Microarray Fabrication through Gene Synthesis (PAGES).

The high genetic variability of RNA viruses is a significant factor limiting the discovery of effective biomarkers, the development of vaccines, and characterizations of the immune response during infection. Protein microarrays have been shown to be a powerful method in biomarker discovery and the identification of novel protein-protein interaction networks, suggesting that this technique could also be very useful in studies of infectious RNA viruses. However, to date, the amount of genetic material required to produce protein arrays, as well as the time- and labor-intensive procedures typically needed, have limited their more widespread application. Here, we introduce a method, protein microarray fabrication through gene synthesis (PAGES), for the rapid and efficient construction of protein microarrays particularly for RNA viruses. Using dengue virus as an example, we first identify consensus sequences from 3,604 different strains and then fabricate complete proteomic microarrays that are unique for each consensus sequence. To demonstrate their applicability, we show that these microarrays can differentiate sera from patients infected by dengue virus, related pathogens, or from uninfected patients. We anticipate that the microarray and expression library constructed in this study will find immediate use in further studies of dengue virus and that, more generally, PAGES will become a widely applied method in the clinical characterization of RNA viruses.

Predictors of Quality of Life in Montreal, Canada: A Longitudinal Study.

This study sought to assess factors associated with quality of life (QoL), and predictive of improvements in QoL over time, in a population-based cohort study. A 4-year longitudinal survey was administered to 2433 individuals at the study baseline; of these, 1828 individuals participated in Wave 2, and 1303 participated in Wave 3. QoL was measured by the Satisfaction with Life Domains Scale. Thirty-two variables were correlated with baseline QoL and together explained 58.2% of the variance. Eleven variables were independent predictors of improvement in QoL over time. Among these variables, social support and stress/coping showed the strongest association with QoL, and neighbourhood characteristics had an additional influence. Multidimensional modelling of a broad spectrum of the factors related to QoL enabled situating mental health and well-being in an ecological system with attendant implications for public health and social policy intervention to facilitate improvement of QoL in the population.