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Background: Inequity in healthcare utilisation refers to differences between groups that remain after adjustment for need for health care. To our knowledge, no previous studies have aimed to assess social inequity in chiropractic utilisation in a general population. Therefore, the objective of this study was to evaluate social inequity in chiropractic utilisation in the general Danish population adjusted for health status as a proxy of need for chiropractic care., Methods: A population-based repeated cross-sectional study design was used based on the Danish National Health Survey in 2010 and 2017. Overall, we included 288,099 individuals aged 30 years or older in 2010 or 2017. For each individual, information on chiropractic utilisation, socioeconomic status, and health status as a proxy of need for chiropractic care was retrieved from nationwide registers using the unique personal identification number. Measures of health status included demographics, poor self-rated physical health, activity limitations, musculoskeletal pain, number of musculoskeletal conditions, and number of chronic diseases. We investigated social inequity in chiropractic utilisation (yes, no) using logistic regression adjusted for health status, stratified by sex and year. Three characteristics of socioeconomic status (educational level, employment status and income) were investigated. To further quantify the degree of social inequity in chiropractic utilisation, we estimated the concentration index of inequity for each of the three characteristics of socioeconomic status., Results: We found significantly higher odds of chiropractic utilisation among individuals with short or medium/long education compared with individuals with elementary education, and among employed individuals compared with individuals who were unemployed, receiving disability pension or retired. Furthermore, the odds of chiropractic utilisation increased with higher income. The concentration index indicated social inequity in chiropractic utilisation in favour of individuals with higher socioeconomic status, with income and employment status contributing more to inequity than educational level., Conclusion: The study demonstrated social inequity in chiropractic utilisation in Denmark beyond differences in health status as a proxy of need for chiropractic care in the general population. The results suggest that new strategies are required if equal treatment for equal need is the goal., (© 2024. The Author(s).)

During intestinal organogenesis, equipotent epithelial progenitors mature into phenotypically distinct stem cells that are responsible for lifelong maintenance of the tissue. While the morphological changes associated with the transition are well characterized, the molecular mechanisms underpinning the maturation process are not fully understood. Here, we leverage intestinal organoid cultures to profile transcriptional, chromatin accessibility, DNA methylation, and three-dimensional (3D) chromatin conformation landscapes in fetal and adult epithelial cells. We observed prominent differences in gene expression and enhancer activity, which are accompanied by local changes in 3D organization, DNA accessibility, and methylation between the two cellular states. Using integrative analyses, we identified sustained Yes-Associated Protein (YAP) transcriptional activity as a major gatekeeper of the immature fetal state. We found the YAP-associated transcriptional network to be regulated at various levels of chromatin organization and likely to be coordinated by changes in extracellular matrix composition. Together, our work highlights the value of unbiased profiling of regulatory landscapes for the identification of key mechanisms underlying tissue maturation.

Background and Aims: Floorball training induces positive effects on health among untrained older adults. However, the effect of long-term participation (>2 years) in floorball training among elderly males has not been investigated. The aim of the present study was to examine the effect of 5 years of floorball training on risk factors for lifestyle diseases, fitness, physical function, and social capital of elderly males and compare to a control group that continued their usual lifestyle., Methods: Twenty-nine recreationally active elderly men aged 75.1 ± 3.3 (mean ± SD; range: 69-81) years with a height, body mass, and body mass index of 1.78 ± 0.06 m, 79.8 ± 10.9 kg, and 25.8 ± 4.1 kg/m 2 , respectively, volunteered to take part in follow-up investigations about 5 years after participating in a study on the effect of 12 weeks of floorball or petanque training. At the end of the parental study 15 subjects chose to participate in floorball training (floorball group [FG]) whereas 14 subjects (control group [CG]), resumed their usual lifestyle. FG participated in small-sided floorball training 1 h ~1.75 times/week for 5 years in a local sports club., Results: From baseline to 5 years, FG had reduced fat percentage, android, and visceral fat, increased total and leg bone mineral density, leg extension maximal voluntary contraction, maximal walk distance in 6 min and 30 s sit-to-stand repetitions, decreased time for 5 sit-to-stand repetitions and Timed Up and Go ( p  < 0.05). These changes were all different from less favorable changes in CG ( p  < 0.05). In FG there was a decline in maximum oxygen uptake which was smaller than the decline in CG ( p  < 0.05). In addition, FG had developed social capital through the 5 years strengthening their social connectedness and group cohesion., Conclusion: In conclusion, both from a sociological and physiological perspective, small-sided floorball training can be considered a health-promoting activity for older men., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors. Health Science Reports published by Wiley Periodicals LLC.)

Organs are anatomically compartmentalised to cater for specialised functions. In the small intestine (SI), regionalisation enables sequential processing of food and nutrient absorption. While several studies indicate the critical importance of non-epithelial cells during development and homeostasis, the extent to which these cells contribute to regionalisation during morphogenesis remains unexplored. Here, we identify a mesenchymal-epithelial crosstalk that shapes the developing SI during late morphogenesis. We find that subepithelial mesenchymal cells are characterised by gradients of factors supporting Wnt signalling and stimulate epithelial growth in vitro. Such a gradient impacts epithelial gene expression and regional villus formation along the anterior-posterior axis of the SI. Notably, we further provide evidence that Wnt signalling directly regulates epithelial expression of Sonic Hedgehog (SHH), which, in turn, acts on mesenchymal cells to drive villi formation. Taken together our results uncover a mechanistic link between Wnt and Hedgehog signalling across different cellular compartments that is central for anterior-posterior regionalisation and correct formation of the SI., (© 2022. The Author(s).)

Background: The COVID-19 pandemic and its associated national lockdowns have been linked to deteriorations in mental health worldwide. A number of studies analysed changes in mental health indicators during the pandemic; however, these studies generally had a small number of timepoints, and focused on the initial months of the pandemic. Furthermore, most studies followed-up the same individuals, resulting in significant loss to follow-up and biased estimates of mental health and its change. Here we report on time trends in key mental health indicators amongst Danish adults over the course of the pandemic (March 2020 - July 2021) focusing on subgroups defined by gender, age, and self-reported previously diagnosed chronic and/or mental illness., Methods: We used time-series data collected by Epinion (N=8,261) with 43 timepoints between 20 March 2020 and 22 July 2021. Using a repeated cross-sectional study design, independent sets of individuals were asked to respond to the Copenhagen Corona-Related Mental Health questionnaire at each timepoint, and data was weighted to population proportions. The six mental health indicators examined were loneliness, anxiety, social isolation, quality of life, COVID-19-related worries, and the mental health scale. Gender, age, and the presence of previously diagnosed mental and/or chronic illness were used to stratify the population into subgroups for comparisons., Results: Poorer mental health were observed during the strictest phases of the lockdowns, whereas better outcomes occurred during reopening phases. Women, young individuals (<34 yrs), and those with a mental- and/or chronic illness demonstrated poorer mean time-series than others. Those with a pre-existing mental illness further had a less reactive mental health time-series. The greatest differences between women/men and younger/older age groups were observed during the second lockdown., Conclusions: People with mental illness have reported disadvantageous but stable levels of mental health indicators during the pandemic thus far, and they seem to be less affected by the factors that result in fluctuating time-series in other subgroups., (© 2022. The Author(s).)

Background: Intestinal stem cell transplantation has been shown to promote mucosal healing and to engender fully functional epithelium in experimental colitis. Hence, stem cell therapies may provide an innovative approach to accomplish mucosal healing in patients with debilitating conditions such as inflammatory bowel disease. However, an approach to label and trace transplanted cells, in order to assess engraftment efficiency and to monitor wound healing, is a key hurdle to overcome prior to initiating human studies. Genetic engineering is commonly employed in animal studies, but may be problematic in humans due to potential off-target and long-term adverse effects., Methods: We investigated the applicability of a panel of fluorescent dyes and nanoparticles to label intestinal organoids for visualization using the clinically approved imaging modality, confocal laser endomicroscopy (CLE). Staining homogeneity, durability, cell viability, differentiation capacity, and organoid forming efficiency were evaluated, together with visualization of labeled organoids in vitro and ex vivo using CLE., Results: 5-Chloromethylfluorescein diacetate (CMFDA) proved to be suitable as it efficiently stained all organoids without transfer to unstained organoids in co-cultures. No noticeable adverse effects on viability, organoid growth, or stem cell differentiation capacity were observed, although single-cell reseeding revealed a dose-dependent reduction in organoid forming efficiency. Labeled organoids were easily identified in vitro using CLE for a duration of at least 3 days and could additionally be detected ex vivo following transplantation into murine experimental colitis., Conclusions: It is highly feasible to use fluorescent dye-based labeling in combination with CLE to trace intestinal organoids following transplantation to confirm implantation at the intestinal target site.

The epigenetic regulator TET2 is frequently mutated in hematological diseases. Mutations have been shown to arise in hematopoietic stem cells early in disease development and lead to altered DNA methylation landscapes and an increased risk of hematopoietic malignancy. Here, we show by genome-wide mapping of TET2 binding sites in different cell types that TET2 localizes to regions of open chromatin and cell-type-specific enhancers. We find that deletion of Tet2 in native hematopoiesis as well as fully transformed acute myeloid leukemia (AML) results in changes in transcription factor (TF) activity within these regions, and we provide evidence that loss of TET2 leads to attenuation of chromatin binding of members of the basic helix-loop-helix (bHLH) TF family. Together, these findings demonstrate that TET2 activity shapes the local chromatin environment at enhancers to facilitate TF binding and provides an example of how epigenetic dysregulation can affect gene expression patterns and drive disease development., (© 2019 Rasmussen et al.; Published by Cold Spring Harbor Laboratory Press.)

Background: Floorball training offers a motivating and socially stimulating team activity for older adults, and 12 weeks of floorball training twice a week among men aged 65-76 years have been shown to have positive effects on a number of physiological parameters important for health. However, the effect of long-term participation in floorball training among male elderly has not been investigated. The aim of the present study was to examine the effect of 26-month self-organized regular participation in floorball training on cardiovascular fitness, body composition, blood lipids, glucose control, and physical function among recreationally active men aged 66-78 years., Methods: After completing a 12-week randomized and controlled intervention with floorball and petanque training in the autumn 2014 or spring 2015, 15 subjects chose to participate in floorball training (floorball group, FG), whereas 16 subjects resumed their usual lifestyle (control group, CG). FG took part in self-organized floorball training 1.7 sessions of 40 min/week, and CG continued their normal recreationally active lifestyle during a 26-month follow-up period. At baseline and after the follow-up period subjects were tested for cardiovascular fitness, glucose control (resting blood samples), body composition dual-energy X-ray absorptiometry (DXA-scanning), and functional capacity., Results: In FG, the decline in maximal oxygen uptake (VO 2max ) during the follow-up period was lower (242 ± 379 mL/min, p  = 0.01), blood glycosylated hemoglobin (HbA1c) increased less (-1.6 ± 2.9 mmol/L, p  = 0.02), and leg bone mineral density increased more (0.03 ± 0.05 g/cm 2 , p  = 0.02) than those in CG. The effects on body mass, total lean body mass, fat mass, blood lipids, and physical function were similar in FG and CG., Conclusion: Approximately twice weekly floorball sessions with 40 min/session over 26-month appear to reduce age-related decline in cardiovascular fitness and glucose control and improve leg bone mineral density, suggesting that long-term participation in floorball training can be considered as a health-enhancing activity in recreationally active male elderly.

Purpose: The aim of the present study was to investigate whether street basketball organized as 3 v 3 on either a half court (HC) with 1 basket or a full court (FC) with 2 baskets could improve fitness and health profiles of untrained men after 3 months of supervised training., Methods: Thirty-five untrained men (aged 20-42 years) completed the pre- and post-intervention testing (FC: n  = 13, HC: n  = 12, CO (control): n  = 10). The training attendance was 2.0 ± 0.4 and 1.9 ± 0.3 times per week in FC and HC, respectively. Mean heart rate (HR) was 83.8 ± 6.0 percent of maximal heart rate (%HR max ) and 84.5 ± 2.9 %HR max in FC and HC, respectively., Results: The 3 months of street basketball training on an FC with 2 baskets increased maximal oxygen uptake (2.4 mL/min/kg (95% confidence interval (CI): 1.0-3.9)), time to exhaustion (47 s (95%CI: 26-67)), lean body mass (0.8 kg (95%CI: 0.1-1.5)), and bone mineral density (0.021 g/cm 2 (95%CI: 0.011-0.031)), whereas mean arterial pressure (-5.6 mmHg (95%CI: -7.5 to 3.7)), body fat percentage (-1.6%, (95%CI: -2.5 to -0.7)), heart rate (-18 bpm (95%CI: -24 to -12)), and blood lactate (median: -1.4 mmol/L (interquartile range: -1.5 to -0.6)) during submaximal running were lowered. The changes were less pronounced after the training period when playing on an HC with 1 basket, but increases in maximal oxygen uptake (1.6 mL/min/kg (95%CI: -0.1 to 3.3)), time to exhaustion (28 s (95%CI: 9-47)), lean body mass (1.3 kg (95%CI: 0.3-2.4)), and lower body fat percentage (-0.9% (95%CI: -1.9 to -0.1)) were observed in this group., Conclusion: Three months of 3 v 3 street basketball training improved fitness and led to broad-spectrum improvements in variables related to overall health profile, with the most marked effects observed when playing on an FC with 2 baskets.

Tissue regeneration requires dynamic cellular adaptation to the wound environment. It is currently unclear how this is orchestrated at the cellular level and how cell fate is affected by severe tissue damage. Here we dissect cell fate transitions during colonic regeneration in a mouse dextran sulfate sodium (DSS) colitis model, and we demonstrate that the epithelium is transiently reprogrammed into a primitive state. This is characterized by de novo expression of fetal markers as well as suppression of markers for adult stem and differentiated cells. The fate change is orchestrated by remodeling the extracellular matrix (ECM), increased FAK/Src signaling, and ultimately YAP/TAZ activation. In a defined cell culture system recapitulating the extracellular matrix remodeling observed in vivo, we show that a collagen 3D matrix supplemented with Wnt ligands is sufficient to sustain endogenous YAP/TAZ and induce conversion of cell fate. This provides a simple model for tissue regeneration, implicating cellular reprogramming as an essential element., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

The effect of small-sided team sport training and protein intake on muscle mass, physical function, and adaptations important for health in untrained older adults was examined. Forty-eight untrained older (72±6 (±standard deviation, SD) years men and women were divided into either a team sport group ingesting a drink high in protein (18 g) immediately and 3 h after each training session (TS-HP, n = 13), a team sport group ingesting an isocaloric drink with low protein content (3 g; TS-LP, n = 18), or a control group continuing their normal activities (CON, n = 17). The team sport training was performed as ~20 min of small-sided ball games twice a week over 12 weeks. After the intervention period, leg muscle mass was 0.6 kg higher (P = 0.047) in TS-HP, with no effect in TS-LP. In TS-HP, number of sit-to-stand repetitions increased (1.2±0.6, P = 0.054), time to perform 2.45 m up-and-go was lower (0.7±0.3 s, P = 0.03) and number of arm curl repetitions increased (3.5±1.2, P = 0.01), whereas in TS-LP only number of repetitions in sit-to-stand was higher (1.6±0.6, P = 0.01). In TS-LP, reductions were observed in total and abdominal fat mass (1.2±0.5 and 0.4±0.2 kg, P = 0.03 and P = 0.02, respectively), heart rate at rest (9±3 bpm, P = 0.002) and plasma C-reactive protein (1.8±0.8 mmol/L, P = 0.03), with no effects in TS-HP. Thus, team sport training improves functional capacity of untrained older adults and increases leg muscle mass only when ingesting proteins after training. Furthermore, team sport training followed by intake of drink with low protein content does lower fat mass, heart rate at rest and level of systemic inflammation., Trial Registration: clinicaltrials.gov NCT03120143.

The treatment of common bile duct (CBD) disorders, such as biliary atresia or ischemic strictures, is restricted by the lack of biliary tissue from healthy donors suitable for surgical reconstruction. Here we report a new method for the isolation and propagation of human cholangiocytes from the extrahepatic biliary tree in the form of extrahepatic cholangiocyte organoids (ECOs) for regenerative medicine applications. The resulting ECOs closely resemble primary cholangiocytes in terms of their transcriptomic profile and functional properties. We explore the regenerative potential of these organoids in vivo and demonstrate that ECOs self-organize into bile duct-like tubes expressing biliary markers following transplantation under the kidney capsule of immunocompromised mice. In addition, when seeded on biodegradable scaffolds, ECOs form tissue-like structures retaining biliary characteristics. The resulting bioengineered tissue can reconstruct the gallbladder wall and repair the biliary epithelium following transplantation into a mouse model of injury. Furthermore, bioengineered artificial ducts can replace the native CBD, with no evidence of cholestasis or occlusion of the lumen. In conclusion, ECOs can successfully reconstruct the biliary tree, providing proof of principle for organ regeneration using human primary cholangiocytes expanded in vitro.

Aims: In vitro studies have demonstrated that formation of reactive oxygen species (ROS) contributes to the effect of bactericidal antibiotics. The formation of ROS is not restricted to bacteria, but also occurs in mammalian cells. Oxidative stress is linked to several diseases. This study investigates whether antibiotic drugs induce oxidative stress in healthy humans as a possible mechanism for adverse reactions to the antibiotic drugs., Methods: This study contains information from two randomised, controlled trials. Participants underwent 1 week treatment with clarithromycin, trimethoprim, phenoxymethylpenicillin (penicillin V), or placebo. Oxidative modifications were measured as 24-h urinary excretion of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) and 8-oxo-7,8-dihydroguanosine (8-oxoGuo), and plasma levels of malondialdehyde before and after treatment as a measurement of DNA oxidation, RNA oxidation, and lipid peroxidation, respectively., Results: Clarithromycin significantly increased urinary excretion of 8-oxodG by 22.0% (95% confidence interval (CI): 3.6-40.4%) and 8-oxoGuo by 14.9% (95% CI: 3.7-26.1%). Further, we demonstrated that trimethoprim significantly lowered urinary excretion of 8-oxodG by 21.7% (95% CI: 5.8-37.6%), but did not influence urinary excretion of 8-oxoGuo. Penicillin V did not influence urinary excretion of 8-oxodG or 8-oxoGuo. None of the antibiotic drugs influenced plasma levels of malondialdehyde., Conclusion: Clarithromycin significantly increases oxidative nucleic acid modifications. Increased oxidative modifications might explain some of clarithromycin's known adverse reactions. Trimethoprim significantly lowers DNA oxidation but not RNA oxidation. Penicillin V had no effect on oxidative nucleic acid modifications., (© 2017 The British Pharmacological Society.)

This article presents the results of a multidisciplinary study which investigated the effects of a period with floorball training on health status, psychological health and social capital of older men. Thirty-nine untrained men aged 69.9 ± 0.6 (range: 65-76) were randomized into a group playing floorball (n = 22) or a group playing petanque (n = 17) one hour twice a week for 12 weeks. Both groups filled out the Health Survey Short Form (SF-12) and the Hospital Anxiety and Depression Scale (HADS) before and after the 12-week intervention. Linear regression analyses with bootstrapping showed that the men in the floorball group improved in the SF-12 composite score for mental health, as well as the HADS subscales anxiety and depression, compared to the men in the petanque group. In addition, 21 interviews were conducted with a sample of the men engaged in floorball. According to the statements in the interviews, the men in the floorball group experienced a high degree of solidarity and group cohesion which seemed to have increased their social capital during the intervention. In particular, the fun and joyful experiences of playing led to a high degree of social connectedness, which were mentioned by many of the men as the main reason for their participation throughout the 12-week period. The statistical results and the interview findings suggest that participation in a ball game such as floorball has several benefits regarding health status, psychological health and social capital and in addition that playing floorball is experienced as enjoyable amongst older men. Thus, it can be concluded that floorball is an activity that benefits older men and should be provided in relevant contexts, such as e.g. sport clubs or centres for seniors., Competing Interests: Conflict of Interest: All authors declare no conflicts of interest in this paper.

Background: Children with cancer experience severe reductions in physical fitness and functionality during and following intensive treatment. This may negatively impact their quality of life., Purpose: To describe the physical capacity and functionality of children with cancer during and after treatment as well as the feasibility of physical activity intervention in the Rehabilitation including Social and Physical activity and Education in Children and Teenagers with Cancer study., Patients and Methods: The study included children diagnosed from January 2013 to April 2016 with paediatric cancer or Langerhans cell histiocytosis, all treated with chemotherapy. Seventy-five of 78 consecutively eligible children (96.2%) were included. Median age was 11 years (range 6‒18). The physical capacity and function were assessed based on testing of physical strength, balance and cardiorespiratory fitness. Children were tested at diagnosis, 3 and 6 months after diagnosis and 1 year after cessation of treatment. The feasibility evaluation was inspired by the criteria for reporting the development and evaluation of complex interventions in healthcare., Results: All children participated in the physical intervention programme with no dropouts. Strenuous physical exercise and physiological testing during paediatric cancer treatment was safe and feasible, with only five minor adverse events during the intervention. Cardiorespiratory fitness was significantly lower in children with cancer than norms for healthy age-matched children at diagnosis (difference 19.1 mL/kg/min, 95% CI 15.4 to 22.7; p <0.0001), during treatment 3 and 6 months from diagnosis (difference 21.0 mL/kg/min, 95% CI 17.4 to 24.6; p <0.0001 and difference 21.6 mL/kg/min, 95% CI 17.3 to 25.8; p <0.0001, respectively) and 1 year after cessation of treatment (difference 6.9 mL/kg/min, 95% CI 1.1 to 12.7; p <0.0072). Furthermore, children with cancer experienced a pronounced decline in physical function., Conclusion: This study shows that it is safe and feasible to perform strenuous physical exercise and testing during paediatric cancer treatment and that children with cancer have significantly lower physical capacity and functionality than healthy age-matched norms., Trial Registration Number: ClinicalTrials.gov: NCT01772862., Competing Interests: Competing interests: None declared.

Significant improvements can be realized by converting conventional batch processes into continuous ones. The main drivers include reduction of cost and waste, increased safety, and simpler scale-up and tech transfer activities. Re-designing the process layout offers the opportunity to incorporate a set of process analytical technologies (PAT) embraced in the Quality-by-Design (QbD) framework. These tools are used for process state estimation, providing enhanced understanding of the underlying variability in the process impacting quality and yield. This work describes a road map for identifying the best technology to speed-up the development of continuous processes while providing the basis for developing analytical methods for monitoring and controlling the continuous full-scale reaction. The suitability of in-line Raman, FT-infrared (FT-IR), and near-infrared (NIR) spectroscopy for real-time process monitoring was investigated in the production of 1-bromo-2-iodobenzene. The synthesis consists of three consecutive reaction steps including the formation of an unstable diazonium salt intermediate, which is critical to secure high yield and avoid formation of by-products. All spectroscopic methods were able to capture critical information related to the accumulation of the intermediate with very similar accuracy. NIR spectroscopy proved to be satisfactory in terms of performance, ease of installation, full-scale transferability, and stability to very adverse process conditions. As such, in-line NIR was selected to monitor the continuous full-scale production. The quantitative method was developed against theoretical concentration values of the intermediate since representative sampling for off-line reference analysis cannot be achieved. The rapid and reliable analytical system allowed the following: speeding up the design of the continuous process and a better understanding of the manufacturing requirements to ensure optimal yield and avoid unreacted raw materials and by-products in the continuous reactor effluent. Graphical Abstract Using PAT to accelerate the transition to continuous API manufacturing.

Chromatin-associated proteins are essential for the specification and maintenance of cell identity. They exert these functions through modulating and maintaining transcriptional patterns. To elucidate the functions of the Jmjd2 family of H3K9/H3K36 histone demethylases, we generated conditional Jmjd2a/Kdm4a, Jmjd2b/Kdm4b and Jmjd2c/Kdm4c/Gasc1 single, double and triple knockout mouse embryonic stem cells (ESCs). We report that while individual Jmjd2 family members are dispensable for ESC maintenance and embryogenesis, combined deficiency for specifically Jmjd2a and Jmjd2c leads to early embryonic lethality and impaired ESC self-renewal, with spontaneous differentiation towards primitive endoderm under permissive culture conditions. We further show that Jmjd2a and Jmjd2c both localize to H3K4me3-positive promoters, where they have widespread and redundant roles in preventing accumulation of H3K9me3 and H3K36me3. Jmjd2 catalytic activity is required for ESC maintenance, and increased H3K9me3 levels in knockout ESCs compromise the expression of several Jmjd2a/c targets, including genes that are important for ESC self-renewal. Thus, continual removal of H3K9 promoter methylation by Jmjd2 demethylases represents a novel mechanism ensuring transcriptional competence and stability of the pluripotent cell identity., (© 2016 The Authors.)

TEN-ELEVEN-TRANSLOCATION-2 (TET2) and DNA-METHYLTRANSFERASE-3A (DNMT3A), both encoding proteins involved in regulating DNA methylation, are mutated in hematological malignancies affecting both myeloid and lymphoid lineages. We previously reported an association of TET2 and DNMT3A mutations in progenitors of patients with angioimmunoblastic T-cell lymphomas (AITL). Here, we report on the cooperative effect of Tet2 inactivation and DNMT3A mutation affecting arginine 882 (DNMT3A(R882H)) using a murine bone marrow transplantation assay. Five out of eighteen primary recipients developed hematological malignancies with one mouse developing an AITL-like disease, two mice presenting acute myeloid leukemia (AML)-like and two others T-cell acute lymphoblastic leukemia (T-ALL)-like diseases within 6 months following transplantation. Serial transplantations of DNMT3A(R882H) Tet2(-/-) progenitors led to a differentiation bias toward the T-cell compartment, eventually leading to AITL-like disease in 9/12 serially transplanted recipients. Expression profiling suggested that DNMT3A(R882H) Tet2(-/-) T-ALLs resemble those of NOTCH1 mutant. Methylation analysis of DNMT3A(R882H) Tet2(-/-) T-ALLs showed a global increase in DNA methylation affecting tumor suppressor genes and local hypomethylation affecting genes involved in the Notch pathway. Our data confirm the transformation potential of DNMT3A(R882H) Tet2(-/-) progenitors and represent the first cooperative model in mice involving Tet2 inactivation driving lymphoid malignancies.

Acute myeloid leukemias (AMLs) with a rearrangement of the mixed-linage leukemia (MLL) gene are aggressive hematopoietic malignancies. Here, we explored the feasibility of using the H3K9- and H3K36-specific demethylases Jmjd2/Kdm4 as putative drug targets in MLL-AF9 translocated leukemia. Using Jmjd2a, Jmjd2b, and Jmjd2c conditional triple-knockout mice, we show that Jmjd2/Kdm4 activities are required for MLL-AF9 translocated AML in vivo and in vitro. We demonstrate that expression of the interleukin 3 receptor α (Il3ra also known as Cd123) subunit is dependent on Jmjd2/Kdm4 through a mechanism involving removal of H3K9me3 from the promoter of the Il3ra gene. Importantly, ectopic expression of Il3ra in Jmjd2/Kdm4 knockout cells alleviates the requirement of Jmjd2/Kdm4 for the survival of AML cells, showing that Il3ra is a critical downstream target of Jmjd2/Kdm4 in leukemia. These results suggest that the JMJD2/KDM4 proteins are promising drug targets for the treatment of AML., (© 2016 Agger et al.; Published by Cold Spring Harbor Laboratory Press.)

The effects of 52 weeks of soccer or resistance training were investigated in untrained elderly men. The subjects aged 68.1±2.1 yrs were randomised into a soccer (SG; n = 9), a resistance (RG; n = 9) and a control group (CG; n = 8). The subjects in SG and RG, respectively, trained 1.7±0.3 and 1.8±0.3 times weekly on average during the intervention period. Muscle function and body composition were determined before and after 16 and 52 weeks of the intervention period. In SG, BMI was reduced by 1.5% and 3.0% (p<0.05) after 16 and 52 weeks, respectively, unchanged in RG and 2% higher (p<0.05) in CG after 52 weeks of the intervention period. In SG, the response to a glucose tolerance test was 16% lower (p<0.05) after 16 wks, but not after 52 wks, compared to before the intervention period, and unchanged in RG and CG. In SG, superoxide dismutase-2 expression was 59% higher (p<0.05) after 52 wks compared to before the intervention period, and unchanged in RG and CG. In RG, upper body lean mass was 3 and 2% higher (p<0.05) after 16 and 52 wks, respectively, compared to before the intervention period, and unchanged in SG and CG. In RG, Akt-2 expression increased by 28% (p<0.01) and follistatin expression decreased by 38% (p<0.05) during the 52-wk intervention period, and was unchanged in SG and CG. Thus, long-term soccer training reduces BMI and improves anti-oxidative capacity, while long-term resistance training impacts muscle protein enzyme expression and increases lean body mass in elderly men. Trial Registration: ClinicalTrials.gov: NCT01530035.

DNA methylation is tightly regulated throughout mammalian development, and altered DNA methylation patterns are a general hallmark of cancer. The methylcytosine dioxygenase TET2 is frequently mutated in hematological disorders, including acute myeloid leukemia (AML), and has been suggested to protect CG dinucleotide (CpG) islands and promoters from aberrant DNA methylation. In this study, we present a novel Tet2-dependent leukemia mouse model that closely recapitulates gene expression profiles and hallmarks of human AML1-ETO-induced AML. Using this model, we show that the primary effect of Tet2 loss in preleukemic hematopoietic cells is progressive and widespread DNA hypermethylation affecting up to 25% of active enhancer elements. In contrast, CpG island and promoter methylation does not change in a Tet2-dependent manner but increases relative to population doublings. We confirmed this specific enhancer hypermethylation phenotype in human AML patients with TET2 mutations. Analysis of immediate gene expression changes reveals rapid deregulation of a large number of genes implicated in tumorigenesis, including many down-regulated tumor suppressor genes. Hence, we propose that TET2 prevents leukemic transformation by protecting enhancers from aberrant DNA methylation and that it is the combined silencing of several tumor suppressor genes in TET2 mutated hematopoietic cells that contributes to increased stem cell proliferation and leukemogenesis., (© 2015 Rasmussen et al.; Published by Cold Spring Harbor Laboratory Press.)

We compared the accuracy of five objective methods, including two newly developed methods combining accelerometry and activity type recognition (Acti4), against indirect calorimetry, to estimate total energy expenditure (EE) of different activities in semi-standardized settings. Fourteen participants performed a standardized and semi-standardized protocol including seven daily life activity types, while having their EE measured by indirect calorimetry. Simultaneously, physical activity was quantified by an ActivPAL3, two ActiGraph GT3X+'s and an Actiheart. EE was estimated by the standard ActivPAL3 software (ActivPAL), ActiGraph GT3X+ (ActiGraph) and Actiheart (Actiheart), and by a combination of activity type recognition via Acti4 software and activity counts per minute (CPM) of either a hip- or thigh-worn ActiGraph GT3X+ (AGhip + Acti4 and AGthigh + Acti4). At group level, estimated physical activities EE by Actiheart (MSE = 2.05) and AGthigh + Acti4 (MSE = 0.25) were not significantly different from measured EE by indirect calorimetry, while significantly underestimated by ActiGraph, ActivPAL and AGhip + Acti4. AGthigh + Acti4 and Actiheart explained 77% and 45%, of the individual variations in measured physical activity EE by indirect calorimetry, respectively. This study concludes that combining accelerometer data from a thigh-worn ActiGraph GT3X+ with activity type recognition improved the accuracy of activity specific EE estimation against indirect calorimetry in semi-standardized settings compared to previously validated methods using CPM only.

Purpose: To determine the time-wise effect of specific resistance training on neck pain among industrial technicians with frequent neck pain symptoms., Methods: Secondary analysis of a parallel-group cluster randomized controlled trial of 20 weeks performed at two large industrial production units in Copenhagen, Denmark. Women with neck pain >30 mm VAS (N = 131) were included in the present analysis. The training group (N = 77) performed specific resistance training for the neck/shoulder muscles three times a week, and the control group (N = 54) received advice to stay active. Participants of both groups registered neck pain intensity (0-100 mm VAS) once a week., Results: Neck pain intensity was 55 mm (SD 23) at baseline. There was a significant group by time interaction for neck pain (F-value 2.61, P<0.001, DF = 19). Between-group differences in neck pain reached significance after 4 weeks (11 mm, 95% CI 2 to 20). The time-wise change in pain showed three phases; a rapid decrease in the training group compared with the control group during the initial 7 weeks, a slower decrease in pain during the following weeks (week 8-15), and a plateau during the last weeks (week 16-20). Adherence to training followed a two-phase pattern, i.e. weekly participation rate was between 70-86% during the initial 7 weeks, dropping towards 55-63% during the latter half of the training period., Conclusion: Four weeks of specific resistance training reduced neck pain significantly, but 15 weeks is required to achieve maximal pain reduction. The time-wise change in pain followed a three-phase pattern with a rapid effect during the initial 7 weeks followed by a slower but still positive effect, and finally a plateau from week 15 and onwards. Decreased participation rate may explain the decreased efficacy during the latter phase of the intervention.

The histone demethylase JMJD2C, also known as KDM4C/GASC1, has activity against methylated H3K9 and H3K36 and is amplified and/or overexpressed in human cancers. By the generation of Jmjd2c knockout mice, we demonstrate that loss of Jmjd2c is compatible with cellular proliferation, embryonic stem cell (ESC) self-renewal, and embryonic development. Moreover, we report that JMJD2C localizes to H3K4me3-positive transcription start sites in both primary cells and in the human carcinoma KYSE150 cell line containing an amplification of the JMJD2C locus. Binding is dependent on the double Tudor domain of JMJD2C, which recognizes H3K4me3 but not H4K20me2/me3 in vitro, showing a binding specificity different from that of the double Tudor domains of JMJD2A and JMJD2B. Depletion of JMJD2C in KYSE150 cells has a modest effect on H3K9me3 and H3K36me3 levels but impairs proliferation and leads to deregulated expression of a subset of target genes involved in cell cycle progression. Taking these findings together, we show that JMJD2C is targeted to H3K4me3-positive transcription start sites, where it can contribute to transcriptional regulation, and report that the putative oncogene JMJD2C generally is not required for cellular proliferation or embryonic development.

Objectives: This study investigated long-term effects and implementation processes of workplace strength training for musculoskeletal disorders., Methods: 333 and 140 laboratory technicians from private and public sector companies, respectively, replied to a 3-year follow-up questionnaire subsequent to a 1-year randomized controlled trial (RCT) with high-intensity strength training for prevention and treatment of neck, shoulder, and arm pain. Being a natural experiment, the two participating companies implemented and modified the initial training program in different ways during the subsequent 2 years after the RCT., Results: At 3-year follow-up the pain reduction in neck, shoulder, elbow, and wrist achieved during the first year was largely maintained at both companies. However, the private sector company was rated significantly better than the public sector company in (1) training adherence, (2) training culture, that is, relatively more employees trained at the workplace and with colleagues, (3) self-reported health changes, and (4) prevention of neck and wrist pain development among initially pain-free employees., Conclusions: This natural experiment shows that strength training can be implemented successfully at different companies during working hours on a long-term basis with lasting effects on pain in neck, shoulder, and arm.

Objective: To investigate the effect of workplace neck/shoulder strength training with and without regular supervision on neck/shoulder pain and headache among office workers., Method: A 20-week cluster randomized controlled trial among 351 office workers was randomized into three groups: two training groups with the same total amount of planned exercises three times per week (1) with supervision (3WS) throughout the intervention period, (2) with minimal supervision (3MS) only initially, and (3) a reference group (REF). Main outcome is self-reported pain intensity in neck and shoulder (scale 0-9) and headache (scale 0-10)., Results: Intention-to-treat analyses showed a significant decrease in neck pain intensity the last 7 days in 3MS compared with REF: -0.5 ± 0.2 (P < 0.02) and a tendency for 3WS versus REF: -0.4 ± 0.2 (P < 0.07). Intensity of headache the last month decreased in both training groups: 3WS versus REF: -1.1 ± 0.2 (P < 0.001) and 3MS versus REF: -1.1 ± 0.2 (P < 0.001). Additionally, days of headache decreased 1.0 ± 0.5 in 3WS and 1.3 ± 0.5 in 3MS versus REF. There were no differences between the two training groups for any of the variables., Conclusion: Neck/shoulder training at the workplace reduced neck pain and headache among office workers independently of the extent of supervision. This finding has important practical implications for future workplace interventions.

The discovery that the Ten-Eleven Translocation (TET) hydroxylases cause DNA demethylation has fundamentally changed the notion of how DNA methylation is regulated. Clonal analysis of the hematopoetic stem cell compartment suggests that TET2 mutations can be early events in hematologic cancers and recent investigations have shown TET2 mutations in diffuse large B-cell lymphoma. However, the detection rates and the types of TET2 mutations vary, and the relation to global methylation patterns has not been investigated. Here, we show TET2 mutations in 12 of 100 diffuse large B-cell lymphomas with 7% carrying loss-of-function and 5% carrying missense mutations. Genome-wide methylation profiling using 450K Illumina arrays identified 315 differentially methylated genes between TET2 mutated and TET2 wild-type cases. TET2 mutations are primarily associated with hypermethylation within CpG islands (70%; P<0.0001), and at CpG-rich promoters (60%; P<0.0001) of genes involved in hematopoietic differentiation and cellular development. Hypermethylated loci in TET2 mutated samples overlap with the bivalent (H3K27me3/H3K4me3) silencing mark in human embryonic stem cells (P=1.5×10(-30)). Surprisingly, gene expression profiling showed that only 11% of the hypermethylated genes were down-regulated, among which there were several genes previously suggested to be tumor suppressors. A meta-analysis suggested that the 35 hypermethylated and down-regulated genes are associated with the activated B-cell-like type of diffuse large B-cell lymphoma in other studies. In conclusion, our data suggest that TET2 mutations may cause aberrant methylation mainly of genes involved in hematopoietic development, which are silenced but poised for activation in human embryonic stem cells.

Background: Previous studies have shown positive effects of physical exercise at the workplace on musculoskeletal disorders. However, long-term adherence remains a challenge. The present study evaluates long-term adherence and effects of a workplace strength training intervention on back, neck and upper extremity pain among laboratory technicians., Methods: Cluster-randomized controlled trial involving 537 industrial laboratory technicians. Subjects were randomized at the cluster level to one of two groups: training group 1 (TG1, n = 282) performing supervised strength training from February to June 2009 (round one) or training group 2 (TG2, n = 255) performing supervised strength training from August to December 2009 (round two). The outcome measures were changes in self-reported pain intensity (0-9) in the back, neck and upper extremity as well as Disability of the Arm, Shoulder and Hand (DASH, 0-100)., Results: Regular adherence, defined as at least one training session per week, was achieved by around 85% in both groups in the supervised training periods. In the intention-to-treat analyses there were significant group by time effects for pain in the neck, right shoulder, right hand and lower back and DASH-resulting in significant reductions in pain (mean 0.3 to 0.5) and DASH (mean 3.9) in the scheduled training group compared to the reference group. For TG1 there were no significant changes in pain in round two, i.e. they maintained the pain reduction achieved in round one. Subgroup analyses among those with severe pain (> = 3 on a scale of 0-9) showed a significant group by time effect for pain in the neck, right shoulder, upper back and lower back. For these subgroups the pain reduction in response to training ranged from 1.1 to 1.8., Conclusions: Specific strength training at the workplace can lead to significant long-term reductions in spinal and upper extremity pain and DASH. The pain reductions achieved during the intensive training phase with supervision appears to be maintained a half year later.

Objectives: To determine the effect of specific resistance training on forearm pain and work disability in industrial technicians., Design and Setting: Two-armed cluster randomised controlled trial of 20 weeks performed at two industrial production units in Copenhagen, Denmark., Participants: Working-age industrial technicians both with and without pain and disability., Interventions: The training group (n=282) performed specific resistance training for the shoulder, neck and arm muscles three times a week. The control group (n=255) was advised to continue normal physical activity., Outcome: All participants rated forearm pain intensity (Visual Analogue Scale, 0-100 mm) once a week (primary outcome) and replied to a questionnaire on work disability (Disability of the Arm Shoulder and Hand, 0-100) at baseline and follow-up (secondary outcome)., Results: Questionnaires were sent to 854 workers of which 30 (n=282) and 27 (n=255) clusters were randomised to training and control, respectively. Of these, 211 and 237 participants, respectively, responded to the follow-up questionnaire. Intention-to-treat analyses including both individuals with and without pain showed that from baseline to follow-up, pain intensity and work disability decreased more in the training group than in the control group (4-5 on a scale of 0-100, p<0.01-0.001). Among those with pain >30 mm Visual Analogue Scale at baseline (n=54), the OR for complete recovery at follow-up in the training group compared with the control group was 4.6 (95% CI 1.2 to 17.9). Among those with work disability >30 at baseline (n=113), the OR for complete recovery at follow-up in the training group compared with the control group was 6.0 (95% CI 1.8 to 19.8)., Conclusion: Specific resistance training of the shoulder, neck and arm reduces forearm pain and work disability among industrial technicians., Trial Registration Number: NCT01071980.

Background: Although leisure-time physical activity is important for health, adherence to regular exercise is challenging for many adults. The workplace may provide an optimal setting to reach a large proportion of the adult population needing regular physical exercise. This study evaluates the effect of implementing strength training at the workplace on non-specific neck and shoulder pain among industrial workers., Methods: Cluster-randomized controlled trial involving 537 adults from occupations with high prevalence of neck and shoulder pain (industrial production units). Participants were randomized to 20 weeks of high-intensity strength training for the neck and shoulders three times a week (n = 282) or a control group receiving advice to stay physically active (n = 255). The strength training program followed principles of progressive overload and periodization. The primary outcome was changes in self-reported neck and shoulder pain intensity (scale 0-9)., Results: 85% of the participants followed the strength training program on a weekly basis. In the training group compared with the control group, neck pain intensity decreased significantly (-0.6, 95% CI -1.0 to -0.1) and shoulder pain intensity tended to decrease (-0.2, 95% CI -0.5 to 0.1, P = 0.07). For pain-cases at baseline (pain intensity > = 3) the odds ratio - in the training group compared with the control group--for being a non-case at follow-up (pain intensity < 3) was 2.0 (95% CI 1.0 to 4.2) for the neck and 3.9 (95% CI 1.7 to 9.4) for the shoulders., Conclusion: High-intensity strength training relying on principles of progressive overload can be successfully implemented at industrial workplaces, and results in significant reductions of neck and shoulder pain., Trial Registration: NCT01071980.

Enzymes catalysing the methylation of the 5-position of cytosine (mC) have essential roles in regulating gene expression and maintaining cellular identity. Recently, TET1 was found to hydroxylate the methyl group of mC, converting it to 5-hydroxymethyl cytosine (hmC). Here we show that TET1 binds throughout the genome of embryonic stem cells, with the majority of binding sites located at transcription start sites (TSSs) of CpG-rich promoters and within genes. The hmC modification is found in gene bodies and in contrast to mC is also enriched at CpG-rich TSSs. We provide evidence further that TET1 has a role in transcriptional repression. TET1 binds a significant proportion of Polycomb group target genes. Furthermore, TET1 associates and colocalizes with the SIN3A co-repressor complex. We propose that TET1 fine-tunes transcription, opposes aberrant DNA methylation at CpG-rich sequences and thereby contributes to the regulation of DNA methylation fidelity.

Background: Neck and shoulder complaints are common among employees in sedentary occupations characterized by intensive computer use. Specific strength training is a promising type of physical exercise for relieving neck and shoulder pain in office workers. However, the optimal combination of frequency and exercise duration, as well as the importance of exercise supervision, is unknown. The VIMS study investigates in a cluster randomized controlled design the effectiveness of different time wise combinations of specific strength training with identical accumulated volume, and the relevance of training supervision for safe and effective training., Methods/design: A cluster randomized controlled trial of 20 weeks duration where employed office workers are randomized to 1 x 60 min, 3 x 20 min, 9 x 7 min per week of specific strength training with training supervision, to 3 x 20 min per week of specific strength training with a minimal amount of training supervision, or to a reference group without training. A questionnaire will be sent to 2000 employees in jobs characterized by intensive computer work. Employees with cardiovascular disease, trauma, hypertension, or serious chronic disease will be excluded. The main outcome measure is pain in the neck and shoulders at week 20., Trial Registration: The trial is registered at ClinicalTrials.gov, number NCT01027390.

We show that histone deacetylase (HDAC) inhibitors lead to functional expression of MHC class I-related chain A and B (MICA/B) on cancer cells, making them potent targets for natural killer (NK) cell-mediated killing through a NK group 2, member D (NKG2D) restricted mechanism. Blocking either apoptosis or oxidative stress caused by HDAC inhibitor treatment did not affect MICA/B expression, suggesting involvement of a separate signal pathway not directly coupled to induction of cell death. HDAC inhibitor treatment induced glycogen synthase kinase-3 (GSK-3) activity and down-regulation of GSK-3 by small interfering RNA or by different inhibitors showed that GSK-3 activity is essential for the induced MICA/B expression. We thus present evidence that cancer cells which survive the direct induction of cell death by HDAC inhibitors become targets for NKG2D-expressing cells like NK cells, gammadelta T cells, and CD8 T cells.

Sudden, unexpected loading to the trunk has been reported in the literature as a potential cause of low-back disorders. This study's aim was to investigate the effect of "readiness training" on the response to sudden back loading among untrained healthy individuals. The study included 19 participants and 19 matched controls. All were employees at the National Institute of Occupational Health. The participants received ten 45-min training sessions during a 4-week period. The training focused on reactions to a variety of expected and unexpected sudden trunk loadings, including balance and coordination exercises. Before and after the training, all subjects were tested for reaction to sudden trunk loading (SL). This entailed applying a horizontal force of 58 N to the subject's upper back. Elapsed time--measured between SL and stopping--decreased significantly in the training group (from 337 to 311 ms) compared with the control group. The improved stopping time was associated with a changed EMG signal, characterized by an increase in the early parts of the response (up to 225 ms) and a subsequent decrease. EMG onset latency was unaffected by training. This study is apparently one of the first to demonstrate that the response to sudden trunk loading can be improved in healthy subjects without an increase in pre-activation and associated trunk stiffness. In perspective, the results indicate a possibility for a training-induced reduction of the risk of low-back injuries, e.g., in nurses exposed to sudden trunk perturbations during patient handling.