The effects of a combined physical activity and health education program on health knowledge and well-being of socially vulnerable children.

The purpose of the present study was to evaluate the effects of the football-based health education program "11 for Health" on health knowledge and well-being in the setting of a 10-week residential stay in a Danish charity home for socially vulnerable children. Six hundred and fourteen children participated in the study, of which 305 took part in the standard program plus "11 for Health" (SG+: 12.4 ± 1.4 (mean ± SD) years) and 309 took part in the standard program (SG: 12.4 ± 1.6 years). We used a crossover design over two full years. The study examined changes in health knowledge and well-being through questionnaires. Overall health knowledge increased more in SG+ than SG over 10 weeks (6.6 vs. 3.4%, p < 0.05). The overall well-being scores were markedly improved (p < 0.05) by 4.54 ± 7.17 AU in SG+ and 4.51 ± 7.16 AU in SG, with no between-group differences. Specific improvements (p < 0.05) were seen in all well-being subcategories, raising the well-being scores to the national average. In conclusion, a 10-week stay at a Danish charity home, the Christmas Seal Home, markedly affected health knowledge and well-being of socially vulnerable children. When the football-based health education program-11 for Health-was added, there were additional positive effects on health knowledge.

An 11-week school-based "health education through football" programme improves musculoskeletal variables in 10-12-yr-old Danish school children.

Purpose: To investigate the effects of the "11 for Health" programme on musculoskeletal fitness. Methods: A total of 108 Danish children aged 10-12 years participated in the study, with 61 children in the intervention group (IG, 25 girls and 36 boys) and 47 children in the control group (CG, 21 girls and 26 boys). Measurements were conducted before and after an 11-week intervention consisting of twice-weekly 45-min football training sessions for IG or continuation of normal Physical Education program for CG. Whole-body dual X-ray absorptiometry was conducted for evaluation of leg and total bone mineral density as well as bone, muscle and fat mass. Standing Long Jump and Stork balance tests were employed to assess musculoskeletal fitness and postural balance. Results: During the 11-week study period, leg bone mineral density as well as leg lean body mass increased more (p < 0.05) in the intervention group (IG) compared to the control group (CG) (0.021 ± 0.019 vs 0.014 ± 0.018 g/cm2 and 0.51 ± 0.46 vs 0.32 ± 0.35 kg, respectively). Moreover, body fat percentage decreased more in IG than in CG (-0.6 ± 0.1 vs 0.1 ± 0.1 %-points, p < 0.05). No significant between-group differences were found in bone mineral content. Stork balance test performance increased more in IG than in CG (0.5 ± 2.6 vs -1.5 ± 4.4 s, p < 0.05), whereas no between-group differences were found in jump performance. Conclusions: The school-based football programme, 11 for Health, with twice-weekly 45-min training sessions over 11 weeks improves various, but not all evaluated parameters related to musculoskeletal fitness in 10-12-yr-old Danish school children.

HIP Fracture REhabilitation Program for older adults with hip fracture (HIP-REP) based on activity of daily living: a feasibility study.

BACKGROUND: A Rehabilitation Program for older adults with hip fracture (HIP-REP) based on Activity of Daily Living has been developed. The objectives of this study were to assess the feasibility and safety of the HIP-REP program to inform a future randomized controlled trial (RCT). METHODS: A feasibility study Inspired by the Complex-intervention development (Medical Research Council framework phase II) design using quantitative and qualitative research methods were conducted. Eighteen participants (above 65 years) with hip fracture were recruited from the orthopedic wards. The setting was cross sectoral including Copenhagen University Hospital, Herlev and Gentofte and rehabilitation centers in Herlev, Gentofte and Lyngby-Taarbæk municipalities. A cross-sectoral rehabilitation intervention tailored to the needs of older adults with hip fracture highlighting systematic goal setting and strategies focused on activities of daily living was conducted. Pre-defined feasibility criteria: participants recruitment and retention, duration of measuring the outcome, adherence to intervention, and adverse events, along with self-reported outcomes and an objective measurement of performance in activity of daily living. Focus groups were analyzed using a deductive manifest content analysis approach. Descriptive statistical analysis and paired t-tests were performed for assessing change in outcome measures. RESULTS: Recruitment rate was 4.5/month. Outcome measures were performed but length and number of questionnaires were a burden. Thirteen out of eighteen participants completed the study three dropped out and two died. Adherence among the 13 was 100%. Focus group revealed issues regarding coordinating the intervention, ensuring procedural processes across sectors regarding recruitment of participants, and documentation in the database. Participants expressed satisfaction with the intervention and felt safe during intervention. Assessment of Motor and Process Skills showed better increase between (range 0.4 to 1.6) in ADL motor ability measures and better increase between (range 0.4 to 0.7) for process ability. No clear association between outcome improvements and intervention adherence. CONCLUSIONS: The cross-sectoral intervention based on daily activities was feasible and safe for older adults with hip fracture. A future RCT, with an improved recruitment strategy and reduced number of outcome measures will evaluate the effectiveness in improving independence and safety performance of activity of daily living. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT03828240 . Registered on January 29, 2019.

Well-Being, Physical Fitness, and Health Profile of 2,203 Danish Girls Aged 10-12 in Relation to Leisure-time Sports Club Activity-With Special Emphasis on the Five Most Popular Sports.

ABSTRACT: Madsen, M, Larsen, MN, Cyril, R, Møller, TK, Madsen, EE, Ørntoft, C, Lind, RR, Ryom, K, Christiansen, SR, Wikman, J, Elbe, AM, and Krustrup, P. Well-being, physical fitness, and health profile of 2,203 Danish girls aged 10-12 in relation to leisure-time sports club activity-with special emphasis on the five most popular sports. J Strength Cond Res 36(8): 2283-2290, 2022-This study investigated the relationship between leisure-time sports club activities and well-being as well as physical health parameters in 10-12-year-old Danish girls. Two thousand two hundred three girls took part in the study, which included questionnaires on participation in leisure-time sports clubs, well-being, Yo-Yo intermittent recovery level 1 children's test, long jump, balance tests, body composition, blood pressure (BP), and resting heart rate (RHR). Data were analyzed according to whether the girls participated in leisure-time sport and according to the 5 most frequently reported sports. Girls enrolled in leisure-time sports had higher physical well-being (49.3 ± 8.6 vs. 45.2 ± 8.3), psychological well-being (50.4 ± 9.0 vs. 49.4 ± 9.8), experienced more peer and social support (50.2 ± 10.0 vs. 48.9 ± 10.7), and perceived a more positive school environment (52.5 ± 8.0 vs. 50.5 ± 9.3), as well as showing higher Yo-Yo (+39%), long jump (+10%), and balance performance (+15%) than girls not involved in sport clubs. The girls active in sports clubs had higher relative muscle mass (+5%), lower fat percentage (-11%), body mass index (-5%), RHR (-3.4 b·min -1 ), and diastolic BP (-1.4 mm Hg) compared with girls not involved in sport ( p < 0.05). Girls who played soccer showed higher aerobic fitness compared with inactive girls (+67%), dancers (+39%), swimmers (+38%), and gymnasts (+16%). Gymnasts had a lower fat percentage than inactive girls (-19%), team handballers (-10%), swimmers (-12%), and soccer players (-4%). Girls participating in club-based leisure-time sports showed higher well-being and better fitness and health profiles than girls not involved in any sports club activities. Girls involved in soccer had better aerobic fitness and gymnasts a lower fat percentage.

Improved metabolic fitness, but no cardiovascular health effects, of a low-frequency short-term combined exercise programme in 50-70-year-olds with low fitness: A randomized controlled trial.

We evaluated the cardiometabolic effects of a 15-week combined exercise programme, implemented in sports clubs, for 50-70-year-olds with low aerobic fitness. In a randomized controlled trial, 45 participants (26 women) with low fitness were randomly assigned (2:1-ratio) to a training group (TG, n = 30) or inactive control group (CG, n = 15). TG had 15 weeks with one weekly 90-min supervised group-based session in a recreational sports club with combined aerobic exercise and strength training and were encouraged to perform home-based training 30 min/wk. Evaluations of relative VO2max (mLO2/min/kg), blood pressure, resting heart rate (HR), echocardiography, peripheral arterial tonometry, body composition, lipid profile and HbA1c were performed at 0 and 15 wks. Average HR during supervised training was 113 ± 13 bpm (68.6 ± 7.0%HRmax), with 4.3 ± 6.6% spent >90%HRmax. At 15-wk follow-up, intention-to-treat analyses revealed no between-group difference for VO2max/kg (0.4 mLO2/min/kg, 95%CI -0.8-1.5, P = 0.519; -3 mL/min, 95%CI -123-118, P = 0.966) or other cardiovascular outcomes (all P > 0.05). Compared to CG, total fat mass (-1.9 kg; 95%CI -3.2 to -0.5, P = 0.005), total fat percentage (-1.3%, 95%CI -2.2 to -0.3, P = 0.01) and total/HDL cholesterol ratio (P = 0.032) decreased in TG. Regular adherence to supervised training was high (81%), but 0% for home-based exercise. In conclusion, the group-based supervised training was associated with high adherence and moderate exercise intensity, whereas insufficiently supported home-based training was not feasible. Together, 15 wks of combined exercise training did not improve aerobic fitness or affected cardiovascular function in 50-70-yr-olds with low aerobic fitness, whereas some positive effects were observed in metabolic parameters.Highlights Combined exercise training implemented in a sports club elicited moderate aerobic intensity in 50-70-year-old untrained individuals.Supervised group-based training had high adherence whereas unsupported home-based training had very low adherence.15 weeks of low-frequency combined moderate intensity exercise training improved lipid profile and fat mass, but had no effect on cardiovascular fitness.

Implementation facilitation of the "11 for Health in Denmark": A case study in a Danish 5th grade class.

The "11 for Health in Denmark" concept aims to enhance 10-12-year-old schoolchildren's physical activity levels and health knowledge through an 11-week football intervention and has been shown to induce positive effects in numerous areas. However, little is known about the implementation facilitation of this concept in schools. This case study therefore aims to gain a deeper understanding of the implementation facilitation of "11 for Health" in a Danish 5th grade class comprising 22 schoolchildren (12 boys and 10 girls, Mage 11.86±) and one teacher. Data were derived from multiple school situations using photographs and videos, observations, and informal interviews. Five themes were identified: (1) environmental and organizational factors impacting on the implementation; (2) familiarization with the concept for the teacher and schoolchildren; (3) previous clique formation; (4) the central role of the teacher; and (5) the importance of how praise partners are allocated. We found high adherence to the "11 for Health" concept, in which the teacher played a key role by applying an autonomy-supportive style of teaching and using the praise partner concept in a suitable manner. A clique of football-playing boys was a resource, as their praising of classmates encouraged adherence. However, familiarization with the "11 for Health" concept was a challenge and the clique's competitive nature occasionally built barriers to other schoolchildren without football experience, potentially leading to a more controlled form of motivation. Based on the results, we encourage teachers to apply autonomy-supportive teaching when applying the "11 for Health" concept.

Detection of lncRNA by LNA-Based In Situ Hybridization in Paraffin-Embedded Cancer Cell Spheroids.

Cancer cell spheroids are considered important preclinical tools to evaluate the efficacy of new drugs. In cancer cell spheroids, the cells assemble and grow in 3D structures with cell contact interactions that are partly impermeable, which leads to central hypoxia and necrosis. The cell spheroids thus possess several features identified in clinical tumors. Not only will the effect and behavior of therapeutic drugs in 3D cell spheroids be affected more similarly than in cells grown on culture plates, but molecular interactions and signaling pathways in cells are also more likely to mimic the in vivo situation. The monitoring of various biomarkers including lncRNAs in 3D cell spheroids is important to assess a potentially induced phenotype in the cells and the effects of drugs. Specifically, for lncRNAs, in situ localization can be done using locked nucleic acid (LNA) probe technology. Here we present a protocol for preparation of cell spheroids for use in LNA probe-based in situ hybridization to study lncRNA expression in paraffin embedded 3D cancer cell spheroids.

One year of Football Fitness improves L1-L4 BMD, postural balance, and muscle strength in women treated for breast cancer.

PURPOSE: To examine efficacy of 12 months Football Fitness offered twice per week on bone mineral density (BMD), bone turnover markers (BTM), postural balance, muscle strength, and body composition in women treated for early-stage breast cancer (BC). METHODS: Women treated for early-stage BC were randomized to Football Fitness (FFG, n = 46) or control (CON, n = 22) in a 2:1 ratio for 12 months, with assessments performed at baseline, 6 months and 12 months. Outcomes were total body-, lumbar spine- and proximal femur BMD, total body lean and fat mass, leg muscle strength, postural balance, and plasma amino-terminal propeptide of type 1 procollagen (P1NP), osteocalcin, and C-terminal telopeptide of type 1 collagen (CTX). Intention-to-treat (ITT) analyses and per-protocol analyses (≥50% attendance in FFG) were performed using linear mixed models. RESULTS: Participants in FFG completing the 12-month intervention (n = 33) attended 0.8 (SD = 0.4) sessions per week. Intention to treat analysis of mean changes over 12 months showed significant differences (p<.05) in L1-L4 BMD (0.029 g/cm2 , 95%CI: 0.001 to 0.057), leg press strength (7.2 kg, 95%CI: 0.1 to 14.3), and postural balance (-4.3 n need of support, 95%CI: -8.0 to -0.7) favoring FFG compared to CON. In the per-protocol analyses, L1-L4 and trochanter major BMD were improved (p = .012 and .030, respectively) in FFG compared with CON. No differences were observed between groups in BTMs in the ITT or per protocol analyses. CONCLUSION: One year of Football Fitness training may improve L1-L4 BMD, leg muscle strength, and postural balance in women treated for early-stage breast cancer.

Exercise intensity and cardiovascular health outcomes after 12 months of football fitness training in women treated for stage I-III breast cancer: Results from the football fitness After Breast Cancer (ABC) randomized controlled trial.

PURPOSE: To examine the exercise intensity and impact of 12 months of twice-weekly recreational football training on cardiorespiratory fitness (CRF), blood pressure (BP), resting heart rate (HRrest), body fat mass, blood lipids, inflammation, and health-related quality of life in women treated for early-stage breast cancer (BC). METHODS: Sixty-eight women who had received surgery for stage I-III BC and completed adjuvant chemo- and/or radiation therapy within 5 years were randomized in a 2:1 ratio to a Football Fitness group (FFG, n = 46) or a control group (CON, n = 22). Football Fitness sessions comprised a warm-up, drills and 3-4 × 7 min of small-sided games (SSG). Assessments were performed at baseline, 6 months and 12 months. Outcomes were peak oxygen uptake (VO2peak), blood pressure (BP), HRrest, total body fat mass, and circulating plasma lipids and hs-CRP, and the 36-Item Short Form Health Survey (SF36). Intention-to-treat (ITT) analyses were performed using linear mixed models. Data are means with SD or 95% confidence intervals. RESULTS: Adherence to training in participants completing the 12-months follow-up (n = 33) was 47.1% (22.7), and HR during SSG was ≥80% of HRmax for 69.8% (26.5) of total playing time. At baseline, VO2peak was 28.5 (6.4) and 25.6 (5.9) ml O2/kg/min in FFG and CON, respectively, and no significant changes were observed at 6- or 12 months follow-up. Systolic BP (SBP) was 117.1 (16.4) and 116.9 (14.8) mmHg, and diastolic BP (DBP) was 72.0 (11.2) and 72.4 (8.5) mmHg in FFG and CON, respectively, at baseline, and a 9.4 mmHg decrease in SBP in CON at 12 months resulted in a between-group difference at 12 months of 8.7 mmHg (p = .012). Blood lipids and hs-CRP were within the normal range at baseline, and there were no differences in changes between groups over the 12 months. Similarly, no differences between groups were observed in HRrest and body fat mass at 6- and12-months follow-up. A between-group difference in mean changes of 23.5 (0.95-46.11) points in the role-physical domain of the SF36 survey favored FFG at 6 months. CONCLUSION: Football Fitness training is an intense exercise form for women treated for breast cancer, and self-perceived health-related limitations on daily activities were improved after 6 months. However, 1 year of Football Fitness training comprising 1 weekly training session on average did not improve CRF, BP, blood lipids, fat mass, or HRrest. TRIAL REGISTRATION NUMBER: The trial was registered at ClinicalTrials.gov with identifier NCT03284567.

MicroRNA In Situ Hybridization in Paraffin-Embedded Cultured Cells.

MicroRNA-21 (miR-21) is one of the most abundant microRNAs in cancer tissues and is considered a strong prognostic biomarker. In situ hybridization (ISH) analyses using locked nucleic acid (LNA) probes have shown that miR-21 is expressed in stromal fibroblastic cells and in subsets of cancer cells. Image analysis of the miR-21 ISH signal has shown that increased expression estimate is associated with poor prognosis in colon cancer. However, assessment of the ISH signal by image analysis to obtain quantitative estimates has been done in retrospective studies without normalization of the expression estimates to reference parameters. The ISH signal output is sensitive to several experimental parameters, including hybridization temperature, probe concentration, and pretreatment, and therefore improved standardized procedures are warranted. We considered the use of paraffin-embedded cultured cells (PECCs) as reference standards that potentially can accompany staining of clinical cancer samples. We found that the cancer cell lines HT-29, CACO-2, and HeLa cells express miR-21 when measured by ISH, and used those cell lines to obtain PECCs. In this methods chapter we present a fixation and embedding procedure to obtain PECCs suitable for microRNA ISH and a double-fluorescence protocol to stain microRNAs together with protein markers in the PECCs.

Automated One-Double-Z Pair BaseScope™ for CircRNA In Situ Hybridization.

Circular RNAs (circRNAs) are single-stranded RNA, typically exons, connected head to tail by back-splicing. The functions of circRNAs include binding of microRNA, regulation of transcription, regulation of alternative splicing, and modulation of immune response. As for other RNA transcripts their levels vary during development and may also become deregulated during disease progression. Different from linear RNAs, the circRNAs are not susceptible to traditional exonuclease activity and therefore more stable in tissues and blood. This makes the circRNAs an attractive new group of potential biomarkers. Specific detection of circRNAs in situ is challenged by the need to discriminate bona fide circRNAs from the linear precursor forms and splice variants that contain largely overlapping sequences. Knowing the sequence around the splice junction site makes the branched DNA probe technology, BaseScope, suitable for selective detection of unique circRNAs. Here, we present the automated application of BaseScope with a one-double-Z pair probe set designed for the junction of circHIPK3.

Co-Detection of miR-21 and TNF-α mRNA in Budding Cancer Cells in Colorectal Cancer.

MicroRNA-21 (miR-21) is upregulated in many cancers including colon cancers and is a prognostic indicator of recurrence and poor prognosis. In colon cancers, miR-21 is highly expressed in stromal fibroblastic cells and more weakly in a subset of cancer cells, particularly budding cancer cells. Exploration of the expression of inflammatory markers in colon cancers revealed tumor necrosis factor alpha (TNF-α) mRNA expression at the invasive front of colon cancers. Surprisingly, a majority of the TNF-α mRNA expressing cells were found to be cancer cells and not inflammatory cells. Because miR-21 is positively involved in cell survival and TNF-α promotes necrosis, we found it interesting to analyze the presence of miR-21 in areas of TNF-α mRNA expression at the invasive front of colon cancers. For this purpose, we developed an automated procedure for the co-staining of miR-21, TNF-α mRNA and the cancer cell marker cytokeratin based on analysis of frozen colon cancer tissue samples (n = 4) with evident cancer cell budding. In all four cases, TNF-α mRNA was seen in a small subset of cancer cells at the invasive front. Evaluation of miR-21 and TNF-α mRNA expression was performed on digital slides obtained by confocal slide scanning microscopy. Both co-expression and lack of co-expression with miR-21 in the budding cancer cells was noted, suggesting non-correlated expression. miR-21 was more often seen in cancer cells than TNF-α mRNA. In conclusion, we report that miR-21 is not linked to expression of the pro-inflammatory cytokine TNF-α mRNA, but that miR-21 and TNF-α both take part in the cancer expansion at the invasive front of colon cancers. We hypothesize that miR-21 may protect both fibroblasts and cancer cells from cell death directed by TNF-α paracrine and autocrine activity.

Feasibility and Health Effects of a 15-Week Combined Exercise Programme for Sedentary Elderly: A Randomised Controlled Trial.

There is strong evidence that considerable health benefits can be achieved even with small amounts of physical activity. However, getting people to exercise regularly is a major challenge not least in the elderly population. This study investigated the feasibility and physiological health effects of a pragmatic 15-week exercise programme for sedentary elderly. In a single-blind randomised controlled trial, 45 sedentary 60-83-year-olds (25 women, 20 men) were randomly assigned (2:1 ratio) to a training group (TG, n=30) or a control group (CG, n=15). The training in TG consisted of a combination of exercise modalities (i.e., strength, aerobic fitness, stability, and flexibility training) performed once a week as supervised group-based training and a weekly home-based training for 15 weeks. Feasibility outcomes were exercise intensity, adherence, and adverse events. The primary outcome was change in aerobic fitness (VO2max/kg). Adherence was high (81%) for the supervised exercise and low (0%) for the home-based exercise. No acute injuries occurred in TG, but 4 subjects (13%) reported considerable joint pain related to training. Average heart rate (HR) during the supervised training was 104±12 beats/min (69.3±8.0%HRmax), with 3.9±7.3% of training time >90%HRmax. Intention-to-treat analyses revealed no between-group differences for aerobic fitness (P=0.790) or any secondary cardiovascular outcomes at 15-week follow-up (resting HR or blood pressure; P>0.05). Compared to CG, bodyweight (-2.3 kg, 95% CI -4.0 to -7.0; P=0.006), total fat mass (-2.0 kg, 95% CI -3.5 to -0.5; P=0.01), and total fat percentage (-1.6%, 95% CI -2.8 to -0.3; P=0.01) decreased in TG. The group-based supervised training had high adherence and moderate exercise intensity, whereas the home-based training was not feasible in this study population. This exercise programme performed once a week did not improve aerobic fitness. Thus, supervised training with more vigorous intensity control appears advisable. Clinical Study registration number is H-15016951.

Chromogen detection of microRNA in frozen clinical tissue samples using LNA™ probe technology.

Specific chromogen- and fluorescence-based detection of microRNA by in situ hybridization (ISH) in formalin-fixed and paraffin-embedded (FFPE) tissue sections has been facilitated by locked nucleic acid (LNA)-based probe technology and can be performed within a single working day. In the current method paper, we present a similar simple 1-day ISH method developed for cryostat sections obtained from clinical cryo-embedded tissue samples. The presented chromogen-based ISH method does not involve proteolytic pretreatment, which is mandatory for FFPE sections, but still retains a sensitivity level similar to that obtained in FFPE sections. The LNA-based ISH method is not only applicable in situations where only access to cryo-embedded material is possible, but it also has a potential use if combining microRNA ISH with immunohistochemistry in double fluorescence staining with antibodies not being compatible with proteolytic predigestion.

miR-21 Expression in Cancer Cells may Not Predict Resistance to Adjuvant Trastuzumab in Primary Breast Cancer.

Trastuzumab is established as standard care for patients with HER2-positive breast cancer both in the adjuvant and metastatic setting. However, 50% of the patients do not respond to the trastuzumab therapy, and therefore new predictive biomarkers are highly warranted. MicroRNAs (miRs) constitute a new group of biomarkers and their cellular expression can be determined in tumor samples by in situ hybridization (ISH) analysis. miR-21 is highly prevalent and up-regulated in breast cancer and has been linked to drug resistance in clinical and in vitro settings. To determine expression patterns of miR-21 in high-grade breast cancers, we examined miR-21 expression in 22 HER2-positive tumors and 15 HER2-negative high-grade tumors by ISH. The histological examination indicated that patient samples could be divided into three major expression patterns: miR-21 predominantly in tumor stroma, predominantly in cancer cells, or in both stromal and cancer cells. There was no obvious difference between the HER2-positive and HER2-negative tumors in terms of the miR-21 expression patterns and intensities. To explore the possibility that miR-21 expression levels and/or cellular localization could predict resistance to adjuvant trastuzumab in HER2-positive breast cancer patients, we analyzed additional 16 HER2-positive tumors from patients who were treated with trastuzumab in the adjuvant setting. Eight of the 16 patients showed clinical recurrence and were considered resistant. Examination of the miR-21 expression patterns and intensities revealed no association between the miR-21 scores in the cancer cell population (p = 0.69) or the stromal cells population (p = 0.13) and recurrent disease after adjuvant trastuzumab. Thus, our findings show that elevated miR-21 expression does not predict resistance to adjuvant trastuzumab.

Motivational counseling to reduce sitting time: a community-based randomized controlled trial in adults.

BACKGROUND: Sedentary behavior is regarded as a distinct risk factor for cardiometabolic morbidity and mortality, but knowledge of the efficacy of interventions targeting reductions in sedentary behavior is limited. PURPOSE: To investigate the effect of an individualized face-to-face motivational counseling intervention aimed at reducing sitting time. DESIGN: A randomized, controlled, observer-blinded, community-based trial with two parallel groups using open-end randomization with 1:1 allocation. SETTING/PARTICIPANTS: A total of 166 sedentary adults were consecutively recruited from the population-based Health2010 Study. INTERVENTION: Participants were randomized to a control (usual lifestyle) or intervention group with four individual theory-based counseling sessions. MAIN OUTCOME MEASURES: Objectively measured overall sitting time (ActivPAL 3TM, 7 days); secondary measures were breaks in sitting time, anthropometric measures, and cardiometabolic biomarkers, assessed at baseline and after 6 months. Data were collected in 2010-2012 and analyzed in 2013-2014 using repeated measures multiple regression analyses. RESULTS: Ninety-three participants were randomized to the intervention group and 73 to the control group, and 149 completed the study. The intervention group had a mean sitting time decrease of -0.27 hours/day, corresponding to 2.9% of baseline sitting time (hours/day); the control group increased mean sitting time by 0.06 hours/day. The between-group difference in change, -0.32 hours/day (95% CI=-0.87, 0.24, p=0.26), was not statistically significant. Significant differences in change in fasting serum insulin of -5.9 pmol/L (95% CI=-11.4, -0.5, p=0.03); homeostasis model assessment-estimated insulin resistance of -0.28 (95% CI=-0.53, -0.03, p=0.03); and waist circumference of -1.42 cm (95% CI=-2.54, -0.29, p=0.01) were observed in favor of the intervention group. CONCLUSIONS: Although the observed decrease in sitting time was not significant, a community-based, individually tailored, theory-based intervention program aimed at reducing sitting time may be effective for increasing standing and improving cardiometabolic health in sedentary adults. TRIAL REGISTRATION: This study is registered at Clinicaltrials.gov (NCT00289237).

A sensitive alternative for microRNA in situ hybridizations using probes of 2'-O-methyl RNA + LNA.

The use of short, high-affinity probes consisting of a combination of DNA and locked nucleic acid (LNA) has enabled the specific detection of microRNAs (miRNAs) by in situ hybridization (ISH). However, detection of low-copy number miRNAs is still not always possible. Here the authors show that probes consisting of 2'-O-methyl RNAs (2OMe) and LNA at every third base (2:1 ratio), under optimized hybridization conditions, excluding yeast RNA from the hybridization buffer, can provide superior performance in detection of miRNA targets in terms of sensitivity and signal-to-noise ratio compared to DNA + LNA probes. Furthermore, they show that hybridizations can be performed in buffers of 4M urea instead of 50% formamide, thereby yielding an equally specific but nontoxic assay. The use of 2OMe + LNA-based probes and the optimized ISH assay enable simple and fast detection of low-copy number miRNA targets, such as miR-130a in mouse brain.

Subdoligranulum variabile gen. nov., sp. nov. from human feces.

During studies on the microflora of human feces we have isolated a strictly anaerobic, non-spore-forming, Gram-negative staining organism which exhibits a somewhat variable coccus-shaped morphology. Comparative 16S ribosomal RNA gene sequencing studies show the unidentified organism is phylogenetically a member of the Clostridium leptum supra-generic rRNA cluster and displays a close affinity to some rDNA clones derived from human and pig feces. The nearest named relatives of the unidentified isolate corresponded to Faecalibacterium prausnitzii (formerly Fusobacterium prausnitzii) displaying a 16S rRNA sequence divergence of approximately 9%, with Anaerofilum agile and A. pentosovorans the next closest relatives of the unidentified bacterium (sequence divergence approximately 10%). Based on phenotypic and phylogenetic considerations, it is proposed that the unusual coccoid-shaped organism be classified as a new genus and species, Subdoligranulum variabile. The type strain of S. variabile is BI 114(T) (=CCUG 47106(T)=DSM 15176(T)).