DNA binding and bridging by human CtIP in the healthy and diseased states.

The human DNA repair factor CtIP helps to initiate the resection of double-stranded DNA breaks for repair by homologous recombination, in part through its ability to bind and bridge DNA molecules. However, CtIP is a natively disordered protein that bears no apparent similarity to other DNA-binding proteins and so the structural basis for these activities remains unclear. In this work, we have used bulk DNA binding, single molecule tracking, and DNA bridging assays to study wild-type and variant CtIP proteins to better define the DNA binding domains and the effects of mutations associated with inherited human disease. Our work identifies a monomeric DNA-binding domain in the C-terminal region of CtIP. CtIP binds non-specifically to DNA and can diffuse over thousands of nucleotides. CtIP-mediated bridging of distant DNA segments is observed in single-molecule magnetic tweezers experiments. However, we show that binding alone is insufficient for DNA bridging, which also requires tetramerization via the N-terminal domain. Variant CtIP proteins associated with Seckel and Jawad syndromes display impaired DNA binding and bridging activities. The significance of these findings in the context of facilitating DNA break repair is discussed.

Recent insights into eukaryotic double-strand DNA break repair unveiled by single-molecule methods.

Genome integrity and maintenance are essential for the viability of all organisms. A wide variety of DNA damage types have been described, but double-strand breaks (DSBs) stand out as one of the most toxic DNA lesions. Two major pathways account for the repair of DSBs: homologous recombination (HR) and non-homologous end joining (NHEJ). Both pathways involve complex DNA transactions catalyzed by proteins that sequentially or cooperatively work to repair the damage. Single-molecule methods allow visualization of these complex transactions and characterization of the protein:DNA intermediates of DNA repair, ultimately allowing a comprehensive breakdown of the mechanisms underlying each pathway. We review current understanding of the HR and NHEJ responses to DSBs in eukaryotic cells, with a particular emphasis on recent advances through the use of single-molecule techniques.

APLF and long non-coding RNA NIHCOLE promote stable DNA synapsis in non-homologous end joining.

The synapsis of DNA ends is a critical step for the repair of double-strand breaks by non-homologous end joining (NHEJ). This is performed by a multicomponent protein complex assembled around Ku70-Ku80 heterodimers and regulated by accessory factors, including long non-coding RNAs, through poorly understood mechanisms. Here, we use magnetic tweezers to investigate the contributions of core NHEJ proteins and APLF and lncRNA NIHCOLE to DNA synapsis. APLF stabilizes DNA end bridging and, together with Ku70-Ku80, establishes a minimal complex that supports DNA synapsis for several minutes under piconewton forces. We find the C-terminal acidic region of APLF to be critical for bridging. NIHCOLE increases the dwell time of the synapses by Ku70-Ku80 and APLF. This effect is further enhanced by a small and structured RNA domain within NIHCOLE. We propose a model where Ku70-Ku80 can simultaneously bind DNA, APLF, and structured RNAs to promote the stable joining of DNA ends.

Do older workers suffer more workplace injuries? A systematic review.

Aging populations are a dramatically increased worldwide trend, both in developed and developing countries. This study examines the prevalence of fatal and non-fatal work-related injuries between young (<45 years old) and older (≥45 years old) workers. A systematic literature review aimed at examining studies comparing safety outcomes, namely fatal and non-fatal injuries, between older and younger workers. Results show that 50% of the reviewed papers suggest that fatal injuries are suffered mainly by older workers, while the remaining 50% show no differences between older and younger workers. Regarding non-fatal injuries, 49% of the reviewed papers found no relationship between workers' age; 31% found increased age as a protective factor against non-fatal injuries; and 19% showed that older workers had a higher risk of non-fatal injuries than younger ones. This review suggests that older workers experience higher rates of fatal injuries, and younger workers experience higher rates of non-fatal injuries.

CTP promotes efficient ParB-dependent DNA condensation by facilitating one-dimensional diffusion from parS.

Faithful segregation of bacterial chromosomes relies on the ParABS partitioning system and the SMC complex. In this work, we used single-molecule techniques to investigate the role of cytidine triphosphate (CTP) binding and hydrolysis in the critical interaction between centromere-like parS DNA sequences and the ParB CTPase. Using a combined optical tweezers confocal microscope, we observe the specific interaction of ParB with parS directly. Binding around parS is enhanced by the presence of CTP or the non-hydrolysable analogue CTPγS. However, ParB proteins are also detected at a lower density in distal non-specific DNA. This requires the presence of a parS loading site and is prevented by protein roadblocks, consistent with one-dimensional diffusion by a sliding clamp. ParB diffusion on non-specific DNA is corroborated by direct visualization and quantification of movement of individual quantum dot labelled ParB. Magnetic tweezers experiments show that the spreading activity, which has an absolute requirement for CTP binding but not hydrolysis, results in the condensation of parS-containing DNA molecules at low nanomolar protein concentrations.

Long Noncoding RNA NIHCOLE Promotes Ligation Efficiency of DNA Double-Strand Breaks in Hepatocellular Carcinoma.

Long noncoding RNAs (lncRNA) are emerging as key players in cancer as parts of poorly understood molecular mechanisms. Here, we investigated lncRNAs that play a role in hepatocellular carcinoma (HCC) and identified NIHCOLE, a novel lncRNA induced in HCC with oncogenic potential and a role in the ligation efficiency of DNA double-stranded breaks (DSB). NIHCOLE expression was associated with poor prognosis and survival of HCC patients. Depletion of NIHCOLE from HCC cells led to impaired proliferation and increased apoptosis. NIHCOLE deficiency led to accumulation of DNA damage due to a specific decrease in the activity of the nonhomologous end-joining (NHEJ) pathway of DSB repair. DNA damage induction in NIHCOLE-depleted cells further decreased HCC cell growth. NIHCOLE was associated with DSB markers and recruited several molecules of the Ku70/Ku80 heterodimer. Further, NIHCOLE putative structural domains supported stable multimeric complexes formed by several NHEJ factors including Ku70/80, APLF, XRCC4, and DNA ligase IV. NHEJ reconstitution assays showed that NIHCOLE promoted the ligation efficiency of blunt-ended DSBs. Collectively, these data show that NIHCOLE serves as a scaffold and facilitator of NHEJ machinery and confers an advantage to HCC cells, which could be exploited as a targetable vulnerability. SIGNIFICANCE: This study characterizes the role of lncRNA NIHCOLE in DNA repair and cellular fitness in HCC, thus implicating it as a therapeutic target.See related commentary by Barcena-Varela and Lujambio, p. 4899.

Gender inequality and sexual height dimorphism in Chile.

Chile has experienced significant improvements in its economy; thus, a secular trend in height has been observed in its population. Gender equality has also improved hand in hand with active policies addressing the gender gap in several dimensions (work, education, health) and overall economic improvement. This study examined changes in sexual height dimorphism in four samples of Chilean male and female working-age subjects and attempted to establish associations with gender equality and welfare. Sexual height dimorphism was calculated and compared with gender equality and overall welfare indicators between 1955 and 1995. Sexual height dimorphism reduction was seen to be strongly associated with greater gender equality and some general welfare indicators, such as the infant mortality rate. Gross domestic product per capita was not associated with sexual height dimorphism, but it showed significant associations with gender equality indicators. Overall, the gender gap has been reduced in Chile, which can be observed through improvements in gender equality indicators and a reduction in height dimorphism, mainly in areas associated with women's health. However, gender equality is still far behind in terms of female labour participation and women in political power, which require attention and further improvements.

Anthropometric data for wheelchair users: a systematic literature review.

The wheelchair user population has special requirements that should be met. However, existing anthropometric data for this specific population are limited. This article characterizes and compares the available literature on anthropometric data of wheelchair users to allow for better understandings and use of data in future studies. A systematic literature review was conducted, where inclusion and exclusion criteria were defined. Forty-one articles were selected to be included in this review. The results show that there is limited information available on wheelchair users' anthropometrics. There is a lack of consistency between studies, regarding the measurements collected, samples used and methods applied. These methodological issues and differences between studies make it difficult to ensure good comparisons of databases and populations and proper designs of spaces and equipment. This study provides valuable information for future studies that need to use anthropometric data, for research and development of new products targeting wheelchair users.

Educational level and its relationship with body height and popliteal height in Chilean male workers.

A secular trend in body height has been experienced in many nations and populations, hypothesized to be the result of better living conditions. Educational level has been shown to be closely associated with body height. This study examined the changes in body height and popliteal height in a group of adult Chilean male workers by age cohort and the relationship of these with educational level. The body heights and popliteal heights of 1404 male workers from the Valparaíso and Metropolitan regions of Chile were measured in 2016. The sample was grouped by level of education (primary, secondary, technical and university) and age (21-30, 31-40 and 41-50 years). Robust ANOVA and post-hoc analyses using a one-step modified M-estimation of location were conducted based on bootstrap resampling. Both body height and popliteal height increased from the older to the younger age cohort. The largest increase was from the 41-50 to the 21-30 group, with a 1.1% increase in body height and 1.7% increase in popliteal height. When educational level was introduced into the analysis there was a marked increase in both body height and popliteal height for each cohort, but only in primary- and secondary-educated workers. Despite showing an overall increase in body height and popliteal height, younger workers with the highest levels of education showed fewer differences between them than did older workers with less education. The differences were larger in the older than in the younger cohorts. Similarly, this trend was less clear in workers with higher levels of education (technical and university), probably because of a dilution effect caused by increased access to higher education by workers in the lower income quintiles.

A comparison of manual anthropometric measurements with Kinect-based scanned measurements in terms of precision and reliability.

BACKGROUND: Collecting anthropometric data for real-life applications demands a high degree of precision and reliability. It is important to test new equipment that will be used for data collectionOBJECTIVE:Compare two anthropometric data gathering techniques - manual methods and a Kinect-based 3D body scanner - to understand which of them gives more precise and reliable results. METHODS: The data was collected using a measuring tape and a Kinect-based 3D body scanner. It was evaluated in terms of precision by considering the regular and relative Technical Error of Measurement and in terms of reliability by using the Intraclass Correlation Coefficient, Reliability Coefficient, Standard Error of Measurement and Coefficient of Variation. RESULTS: The results obtained showed that both methods presented better results for reliability than for precision. Both methods showed relatively good results for these two variables, however, manual methods had better results for some body measurements. CONCLUSION: Despite being considered sufficiently precise and reliable for certain applications (e.g. apparel industry), the 3D scanner tested showed, for almost every anthropometric measurement, a different result than the manual technique. Many companies design their products based on data obtained from 3D scanners, hence, understanding the precision and reliability of the equipment used is essential to obtain feasible results.

Assessment of the intraday variability of anthropometric measurements in the work environment: a pilot study.

Sitting for long periods of time, both during work and leisure times, is the typical behavior of the modern society. Especially at work, where there is not much flexibility, adopting the sitting posture for the entire day can cause some short-term and long-term effects. As workers' productivity and well-being relies on working conditions, evaluating the effects caused by work postures assumes a very important role. The purpose of this article was to evaluate the variation of some anthropometric measurements during one typical workday to understand whether the known long-term effects can also be seen and quantified in an 8-h period. Twenty participants were measured before and after work, using traditional anthropometry equipment. The data from the two repetitions were compared using statistical tests. The results showed a slight variation in the anthropometric measurements, some with a tendency to increase over time and others with a tendency to decrease.

Insights on the apparel needs and limitations for athletes with disabilities: The design of wheelchair rugby sports-wear.

Wheelchair rugby is a sport that has been gaining popularity with athletes with disabilities. However, as it is relatively new and not played by the masses, market specific sports-wear is not available for this sport, which impacts directly on performance and clothing satisfaction of the athletes. Therefore, the main objective of this research was to identify the problems that wheelchair rugby players face with the sports-wear they use for playing the game. The data was collected using a focus group and a questionnaire with 61 wheelchair rugby players in the United Kingdom. Based on their suggestions, on the team expertise, and on the literature, a set of design recommendations was proposed for the upper body garments (tops), lower body garments (bottoms), and gloves. The results demonstrated that the gloves currently available negatively impact on players' ability to participate with satisfactory levels of protection and comfort. Moreover, tops and bottoms also present issues, mainly in the fit and ability to regulate the core body temperature. Hence, the recommendations proposed can provide designers with key information on the specific sports-wear requirements and allow them to design and develop products that can satisfy real needs of specific end-users. This paper intends to raise awareness of the needs of sports-wear for those playing wheelchair rugby and promote the inclusivity of athletes with disabilities.

Validation study of a Kinect based body imaging system.

BACKGROUND: Understanding the reliability and precision of the data obtained using three-dimensional body scanners is very important if it is intended to replace the traditional data collection methods. If the collection of anthropometric data with three-dimensional body scanners is a fast and reliable process that produces precise data at a low price, it could be used for numerous applications worldwide. Many studies have addressed data collected by white light and laser based scanners. OBJECTIVE: This study provides a comparative analysis between the anthropometric data collected using a Kinect body imaging system with the data collected using traditional manual methods. Moreover, a comparison is also made between the results obtained in this study and the results of previous studies of different types of body scanners. METHODS: The Mean Absolute Difference was calculated and all the values were compared to the maximum allowable error defined in ISO 20685. Additionally, an analysis of the significant differences between the two acquisition methods was also applied to a physical mannequin, to understand how the body movement and body stance variation in human participants impacts the results obtained. RESULTS: There are few body measurements that are close to this restricted allowable error. The results were better when the mannequin was measured. Although they were still above the ISO 20685 limit, they were much closer than the results obtained for human participants. CONCLUSION: The main cause of the differences between the two methods is the time required for the 3D system to acquire the data. The involuntary body sway of human participants is more difficult to control when the time span is too long.