Effect of serum and cumulus cell expansion on marker gene transcripts in bovine cumulus-oocyte complexes during maturation in vitro.

OBJECTIVE: To determine the distribution of transcripts encoding the FSH receptor (FSHr), LH receptor (LHr), connexin 43 (Cx43), cyclooxygenase-2 (COX-2), and prostaglandin E(2) receptors 2 and 3 (EP2 and EP3) within bovine cumulus-oocyte complexes (COCs) and denuded oocytes and investigate the influence of gonadotropins, serum, and cumulus cell expansion on the abundance of transcripts encoding these genes. DESIGN: Prospective controlled animal study. SETTING: University research laboratory. PATIENT(S): Animal models for human studies. INTERVENTION(S): Cumulus-oocyte complexes were treated in culture with serum and gonadotropin-supplemented media to examine the effects to mRNA transcript levels. MAIN OUTCOME MEASURE(S): Variation in mRNA transcript levels. RESULT(S): Luteinizing hormone receptor, FSHr, and EP3 mRNAs were detected in intact COCs and not in cumulus cell-denuded oocytes, whereas Cx43, COX-2, and EP2 mRNAs were found in both COCs and oocytes. The relative abundance of marker gene mRNAs did not vary in media containing no additives or FSH alone, independent of whether the media induced cumulus cell expansion. However, the presence of serum in maturation media significantly decreased expression of all mRNAs except LHr. CONCLUSION(S): The relative abundance of COC mRNAs is altered by serum in the maturation medium, which may signify long-term consequences for embryonic development

The relative influence of demographic, individual, social, and environmental factors on physical activity among boys and girls

Background: This study aimed to evaluate the associations of selected demographic, individual, social, and environmental factors with moderate-to-vigorous physical activity (MVPA) in a sample of children and adolescents. Methods: MVPA was assessed among youth (n = 294) 10-17-years-old using the ActiGraph accelerometer. Youth completed measures of demographic and individual variables related to physical activity (PA), perceived social support by parents and peers, and perceived neighborhood characteristics. Parents completed the long-form of the International Physical Activity Questionnaire. The Physical Activity and Media Inventory was used to measure the home environment and Geographical Information Systems software was used to measure the physical neighborhood environment. Bivariate correlations and hierarchical multiple regression were conducted stratified by gender. Results: Boys participated in significantly more MVPA than girls. In hierarchical analyses, peer support, home PA equipment, and temperature were significantly associated with MVPA among boys whereas distance to the school they attended was associated with MVPA among girls. The final models accounted for 25% and 15% of the variance in MVPA among boys and girls, respectively. Conclusions: Important differences exist among the individual, social, and environmental factors related to MVPA between boys and girls. Boys\u27 levels of activity appear to be influenced by factors closely linked to unstructured and social types of activities whereas girls\u27 activities relate to internal and external barriers as well as their proximity to their schools. The prospective contribution of these important individual, social, and environmental factors to changes in MVPA among children and adolescents remains to be determined

Transgenic mouse model harboring the transcriptional fusion Ccl20-luciferase as a novel reporter of pro-inflammatory response

The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo.Laboratorio de Investigaciones del Sistema InmuneFacultad de Ciencias Exacta

The relative influence of demographic, individual, social, and environmental factors on physical activity among boys and girls

<p>Abstract</p> <p>Background</p> <p>This study aimed to evaluate the associations of selected demographic, individual, social, and environmental factors with moderate-to-vigorous physical activity (MVPA) in a sample of children and adolescents.</p> <p>Methods</p> <p>MVPA was assessed among youth (n = 294) 10-17-years-old using the ActiGraph accelerometer. Youth completed measures of demographic and individual variables related to physical activity (PA), perceived social support by parents and peers, and perceived neighborhood characteristics. Parents completed the long-form of the International Physical Activity Questionnaire. The Physical Activity and Media Inventory was used to measure the home environment and Geographical Information Systems software was used to measure the physical neighborhood environment. Bivariate correlations and hierarchical multiple regression were conducted stratified by gender.</p> <p>Results</p> <p>Boys participated in significantly more MVPA than girls. In hierarchical analyses, peer support, home PA equipment, and temperature were significantly associated with MVPA among boys whereas distance to the school they attended was associated with MVPA among girls. The final models accounted for 25% and 15% of the variance in MVPA among boys and girls, respectively.</p> <p>Conclusions</p> <p>Important differences exist among the individual, social, and environmental factors related to MVPA between boys and girls. Boys' levels of activity appear to be influenced by factors closely linked to unstructured and social types of activities whereas girls' activities relate to internal and external barriers as well as their proximity to their schools. The prospective contribution of these important individual, social, and environmental factors to changes in MVPA among children and adolescents remains to be determined.</p

Impacts of active school design on schooltime sedentary behavior and physical activity: A pilot natural experiment

Background Children spend a significant portion of their days in sedentary behavior (SB) and on average fail to engage in adequate physical activity (PA). The school built environment may influence SB and PA, but research is limited. This natural experiment evaluated whether an elementary school designed to promote movement impacted students\u27 school-time SB and PA. Methods Accelerometers measured SB and PA at pre and post time-points in an intervention group who moved to the new school (n = 21) and in a comparison group experiencing no school environmental change (n = 20). Difference-in-difference (DD) analysis examined SB and PA outcomes in these groups. Measures were also collected post-intervention from an independent, grade-matched group of students in the new school (n = 21). Results As expected, maturational increases in SB were observed. However, DD analysis estimated that the intervention attenuated increase in SB by 81.2 ± 11.4 minutes/day (p\u3c0.001), controlling for time in moderate to vigorous physical activity (MVPA). The intervention was also estimated to increase daily number of breaks from SB by 23.4 ± 2.6 (p \u3c .001) and to increase light physical activity (LPA) by 67.7 ± 10.7 minutes/day (p\u3c0.001). However, the intervention decreased MVPA by 10.3 ± 2.3 minutes/day (p\u3c0.001). Results of gradematched independent samples analysis were similar, with students in the new vs. old school spending 90.5 ± 16.1 fewer minutes/day in SB, taking 21.1 ± 2.7 more breaks from SB (p\u3c0.001), and spending 64.5 ± 14.8 more minutes in LPA (p\u3c0.001), controlling for time in MVPA. Students in the new school spent 13.1 ± 2.7 fewer minutes in MVPA (p\u3c0.001) than their counterparts in the old school. Conclusions This pilot study found that active school design had beneficial effects on SB and LPA, but not on MVPA. Mixed results point to a need for active classroom design strategies to mitigate SB, and quick access from classrooms to areas permissive of high-intensity activities to promote MVPA. Integrating active design with programs/policies to promote PA may yield greatest impact on PA of all intensities

Pattern Transfer of Sub-10 nm Features via Tin-Containing Block Copolymers

Tin-containing block copolymers were investigated as materials for nanolithographic applications. Poly(4-trimethylstannylstyrene-block-styrene) (PSnS-PS) and poly(4-trimethylstannylstyrene-block-4-methoxystyrene) (PSnS-PMOST) synthesized by reversible addition–fragmentation chain transfer polymerization form lamellar domains with periodicities ranging from 18 to 34 nm. Thin film orientation control was achieved by thermal annealing between a neutral surface treatment and a top coat. Incorporation of tin into one block facilitates pattern transfer into SiO_2 via a two-step etch process utilizing oxidative and fluorine-based etch chemistries

Pilot evaluation of a walking school bus program in a low-income, urban community

<p>Abstract</p> <p>Background</p> <p>To evaluate the impact of a walking school bus (WSB) program on student transport in a low-income, urban neighborhood.</p> <p>Methods</p> <p>The design was a controlled, quasi-experimental trial with consecutive cross-sectional assessments. The setting was three urban, socioeconomically disadvantaged, public elementary schools (1 intervention vs. 2 controls) in Seattle, Washington, USA. Participants were ethnically diverse students in kindergarten-5^thgrade (aged 5–11 years). The intervention was a WSB program consisting of a part-time WSB coordinator and parent volunteers. Students' method of transportation to school was assessed by a classroom survey at baseline and one-year follow-up. The Pearson Chi-squared test compared students transported to school at the intervention versus control schools at each time point. Due to multiple testing, we calculated adjusted p-values using the Ryan-Holm stepdown Bonferroni procedure. McNemar's test was used to examine the change from baseline to 12-month follow-up for walking versus all other forms of school transport at the intervention or control schools.</p> <p>Results</p> <p>At baseline, the proportions of students (n = 653) walking to the intervention (20% +/- 2%) or control schools (15% +/- 2%) did not differ (<it>p </it>= 0.39). At 12-month follow up, higher proportions of students (n = 643, <it>p </it>= 0.001)) walked to the intervention (25% +/- 2%) versus the control schools (7% +/- 1%). No significant changes were noted in the proportion of students riding in a car or taking the school bus at baseline or 12-month follow up (all <it>p </it>> 0.05). Comparing baseline to 12-month follow up, the numbers of students who walked to the intervention school increased while the numbers of students who used the other forms of transport did not change (<it>p </it>< 0.0001). In contrast, the numbers of students who walked to the control schools decreased while the numbers of students who used the other forms of transport did not change (<it>p </it>< 0.0001).</p> <p>Conclusion</p> <p>A WSB program is a promising intervention among urban, low-income elementary school students that may promote favorable changes toward active transport to school.</p> <p>Trial Registration</p> <p>ClinicalTrials.gov NCT00402701</p

Clostridium difficile ribotypes in Austria: a multicenter, hospital-based survey

A prospective, noninterventional survey was conducted among Clostridium difficile positive patients identified in the time period of July until October 2012 in 18 hospitals distributed across all nine Austrian provinces. Participating hospitals were asked to send stool samples or isolates from ten successive patients with C.difficile infection to the National Clostridium difficile Reference Laboratory at the Austrian Agency for Health and Food Safety for PCR-ribotyping and in vitro susceptibility testing. A total of 171 eligible patients were identified, including 73 patients with toxin-positive stool specimens and 98 patients from which C. difficile isolates were provided. Of the 159 patients with known age, 127 (74.3 %) were 65 years or older, the median age was 76 years (range: 9–97 years), and the male to female ratio 2.2. Among these patients, 73 % had health care-associated and 20 % community-acquired C. difficile infection (indeterminable 7 %). The all-cause, 30-day mortality was 8.8 % (15/171). Stool samples yielded 46 different PCR-ribotypes, of which ribotypes 027 (20 %), 014 (15.8 %), 053 (10.5 %), 078 (5.3 %), and 002 (4.7 %) were the five most prevalent. Ribotype 027 was found only in the provinces Vienna, Burgenland, and Lower Austria. Severe outcome of C. difficile infection was found to be associated with ribotype 053 (prevalence ratio: 3.04; 95 % CI: 1.24, 7.44), not with the so-called hypervirulent ribotypes 027 and 078. All 027 and 053 isolates exhibited in vitro resistance against moxifloxacin. Fluoroquinolone use in the health care setting must be considered as a factor favoring the spread of these fluoroquinolone resistant C. difficile clones

Transgenic mouse model harboring the transcriptional fusion Ccl20-luciferase as a novel reporter of pro-inflammatory response

The chemokine CCL20, the unique ligand of CCR6 functions as an attractant of immune cells. Expression of CCL20 is induced by Toll-like Receptor (TLR) signaling or proinflammatory cytokine stimulation. However CCL20 is also constitutively produced at specific epithelial sites of mucosa. This expression profile is achieved by transcriptional regulation. In the present work we characterized regulatory features of mouse Ccl20 gene. Transcriptional fusions between the mouse Ccl20 promoter and the firefly luciferase (luc) encoding gene were constructed and assessed in in vitro and in vivo assays. We found that liver CCL20 expression and luciferase activity were upregulated by systemic administration of the TLR5 agonist flagellin. Using shRNA and dominant negative form specific for mouse TLR5, we showed that this expression was controlled by TLR5. To address in situ the regulation of gene activity, a transgenic mouse line harboring a functional Ccl20-luc fusion was generated. The luciferase expression was highly concordant with Ccl20 expression in different tissues. Our data indicate that the transgenic mouse model can be used to monitor activation of innate response in vivo.Laboratorio de Investigaciones del Sistema InmuneFacultad de Ciencias Exacta

A calibration protocol for population-specific accelerometer cut-points in children

PurposeTo test a field-based protocol using intermittent activities representative of children\u27s physical activity behaviours, to generate behaviourally valid, population-specific accelerometer cut-points for sedentary behaviour, moderate, and vigorous physical activity.MethodsTwenty-eight children (46% boys) aged 10&ndash;11 years wore a hip-mounted uniaxial GT1M ActiGraph and engaged in 6 activities representative of children\u27s play. A validated direct observation protocol was used as the criterion measure of physical activity. Receiver Operating Characteristics (ROC) curve analyses were conducted with four semi-structured activities to determine the accelerometer cut-points. To examine classification differences, cut-points were cross-validated with free-play and DVD viewing activities.ResultsCut-points of &le;372, &gt;2160 and &gt;4806 counts&bull;min&minus;1 representing sedentary, moderate and vigorous intensity thresholds, respectively, provided the optimal balance between the related needs for sensitivity (accurately detecting activity) and specificity (limiting misclassification of the activity). Cross-validation data demonstrated that these values yielded the best overall kappa scores (0.97; 0.71; 0.62), and a high classification agreement (98.6%; 89.0%; 87.2%), respectively. Specificity values of 96&ndash;97% showed that the developed cut-points accurately detected physical activity, and sensitivity values (89&ndash;99%) indicated that minutes of activity were seldom incorrectly classified as inactivity.ConclusionThe development of an inexpensive and replicable field-based protocol to generate behaviourally valid and population-specific accelerometer cut-points may improve the classification of physical activity levels in children, which could enhance subsequent intervention and observational studies.<br /