Exercise-induced tendon and bone injury in recreational runners: A test-retest reliability study

Background: Long-distance runners are prone to injuries including Achilles tendinopathy and medial tibial stress syndrome. We have developed an Internet comprehensive self-report questionnaire examining the medical history, injury history, and running habits of adult recreational runners. Objective: The objective of the study was to evaluate two alternative forms of test-retest reliability of a comprehensive self-report Internet questionnaire retrospectively examining the medical history, injury history, and running habits among a sample of adult recreational runners. This will contribute to the broad aims of a wider study investigating genetics and running injury. Methods: Invitations to complete an Internet questionnaire were sent by email to a convenience pilot population (test group 1). Inclusion criteria required participants to be a recreational runner age 18 or over, who ran over 15 km per week on a consistent basis. The survey questions addressed regular running habits and any injuries (including signs, symptoms, and diagnosis) of the lower limbs that resulted in discontinuation of running for a period of 2 consecutive weeks or more, within the last 2 years. Questions also addressed general health, age, sex, height, weight, and ethnic background. Participants were then asked to repeat the survey using the Internet platform again after 10-14 days. Following analysis of test group 1, we soft-launched the survey to a larger population (test group 2), through a local running club of 900 members via email platform. The same inclusion criteria applied, however, participants were asked to complete a repeat of the survey by telephone interview after 7-10 days. Selected key questions, important to clarify inclusion or exclusion from the wider genetics study, were selected to evaluate test-retest reliability. Reliability was quantified using the kappa coefficient for categorical data. Results: In response to the invitation, 28 participants accessed the survey from test group 1, 23 completed the Internet survey on the first occasion, and 20 completed the Internet retest within 10-21 days. Test-retest reliability scored moderate to almost perfect (kappa=.41 to .99) for 19/19 of the key questions analyzed. Following the invitation, 122 participants accessed the survey from test group 2, 101 completed the Internet survey on the first occasion, and 50 were randomly selected and contacted by email inviting them to repeat the survey by telephone interview. There were 33 participants that consented to the telephone interview and 30 completed the questionnaire within 7-10 days. Test-retest reliability scored moderate to almost perfect for 18/19 (kappa=.41 to .99) and slight for 1/19 of the key questions analyzed. Conclusions: We successfully developed a self-reported, retrospective questionnaire, delivered using Internet software, providing stable and reliable answers. We demonstrate that our survey provides a relatively quick, easy to complete, and cost effective method to collect epidemiological data from recreational runners and evaluate these participants for inclusion into a genetic study

Immunofluorescent staining reveals hypermethylation of microchromosomes in the central bearded dragon, Pogona vitticeps

BACKGROUND: Studies of model organisms have demonstrated that DNA cytosine methylation and histone modifications are key regulators of gene expression in biological processes. Comparatively little is known about the presence and distribution of epigenetic marks in non-model amniotes such as non-avian reptiles whose genomes are typically packaged into chromosomes of distinct size classes. Studies of chicken karyotypes have associated the gene-richness and high GC content of microchromosomes with a distinct epigenetic landscape. To determine whether this is likely to be a common feature of amniote microchromosomes, we have analysed the distribution of epigenetic marks using immunofluorescence on metaphase chromosomes of the central bearded dragon (Pogona vitticeps). This study is the first to study the distribution of epigenetic marks on non-avian reptile chromosomes. RESULTS: We observed an enrichment of DNA cytosine methylation, active modifications H3K4me2 and H3K4me3, as well as the repressive mark H3K27me3 in telomeric regions on macro and microchromosomes. Microchromosomes were hypermethylated compared to macrochromosomes, as they are in chicken. However, differences between macro- and microchromosomes for histone modifications associated with actively transcribed or repressed DNA were either less distinct or not detectable. CONCLUSIONS: Hypermethylation of microchromosomes compared to macrochromosomes is a shared feature between P. vitticeps and avian species. The lack of the clear distinction between macro- and microchromosome staining patterns for active and repressive histone modifications makes it difficult to determine at this stage whether microchrosome hypermethylation is correlated with greater gene density as it is in aves, or associated with the greater GC content of P. vitticeps microchromosomes compared to macrochromosomes.This work was supported by a University of Canberra postdoctoral fellowship (awarded to TE, SR, Stephen Sarre, JED, Kris Hardy and Arthur Georges, and supporting RD and AL). TE is supported by an Australian Research Council Future Fellowship (FT110100733)

Immunofluorescent staining reveals hypermethylation of microchromosomes in the central bearded dragon, Pogona vitticeps

BACKGROUND: Studies of model organisms have demonstrated that DNA cytosine methylation and histone modifications are key regulators of gene expression in biological processes. Comparatively little is known about the presence and distribution of epigenetic marks in non-model amniotes such as non-avian reptiles whose genomes are typically packaged into chromosomes of distinct size classes. Studies of chicken karyotypes have associated the gene-richness and high GC content of microchromosomes with a distinct epigenetic landscape. To determine whether this is likely to be a common feature of amniote microchromosomes, we have analysed the distribution of epigenetic marks using immunofluorescence on metaphase chromosomes of the central bearded dragon (Pogona vitticeps). This study is the first to study the distribution of epigenetic marks on non-avian reptile chromosomes. RESULTS: We observed an enrichment of DNA cytosine methylation, active modifications H3K4me2 and H3K4me3, as well as the repressive mark H3K27me3 in telomeric regions on macro and microchromosomes. Microchromosomes were hypermethylated compared to macrochromosomes, as they are in chicken. However, differences between macro- and microchromosomes for histone modifications associated with actively transcribed or repressed DNA were either less distinct or not detectable. CONCLUSIONS: Hypermethylation of microchromosomes compared to macrochromosomes is a shared feature between P. vitticeps and avian species. The lack of the clear distinction between macro- and microchromosome staining patterns for active and repressive histone modifications makes it difficult to determine at this stage whether microchrosome hypermethylation is correlated with greater gene density as it is in aves, or associated with the greater GC content of P. vitticeps microchromosomes compared to macrochromosomes

Multiple roles of H2A.Z in regulating promoter chromatin architecture in human cells

Chromatin accessibility of a promoter is fundamental in regulating transcriptional activity. The histone variant H2A.Z has been shown to contribute to this regulation, but its role has remained poorly understood. Here, we prepare high-depth maps of the position and accessibility of H2A.Z-containing nucleosomes for all human Pol II promoters in epithelial, mesenchymal and isogenic cancer cell lines. We find that, in contrast to the prevailing model, many different types of active and inactive promoter structures are observed that differ in their nucleosome organization and sensitivity to MNase digestion. Key aspects of an active chromatin structure include positioned H2A.Z MNase resistant nucleosomes upstream or downstream of the TSS, and a MNase sensitive nucleosome at the TSS. Furthermore, the loss of H2A.Z leads to a dramatic increase in the accessibility of transcription factor binding sites. Collectively, these results suggest that H2A.Z has multiple and distinct roles in regulating gene expression dependent upon its location in a promoter.This work was supported by grants from the National Institutes of Health, R01 DA033775 (J.H.D.); The FSU Foundation Developing Scholar Award, DSA040358 (J.H.D.); and the National Health and Medical Research Council, 1104340 (D.J.T.)

The Histone Variant H2A.Z Is a Master Regulator of the Epithelial-Mesenchymal Transition

Summary: Epithelial-mesenchymal transition (EMT) is a profound example of cell plasticity that is crucial for embryonic development and cancer. Although it has long been suspected that chromatin-based mechanisms play a role in this process, no master regulator that can specifically regulate EMT has been identified to date. Here, we show that H2A.Z can coordinate EMT by serving as either an activator or repressor of epithelial or mesenchymal gene expression, respectively. Following induction of EMT by TGF-β, we observed an unexpected loss of H2A.Z across both downregulated epithelial and upregulated mesenchymal promoters. Strikingly, the repression of epithelial gene expression was associated with reduction of H2A.Z upstream of the transcription start site (TSS), while the activation of mesenchymal gene expression was dependent on removal of H2A.Z downstream of the TSS. Therefore, the ability of H2A.Z to regulate EMT is dependent on its position, either upstream or downstream of the TSS. : EMT is one of the most intensely studied differentiation-dedifferentiation processes. Domaschenz et al. now demonstrate that H2A.Z has the unique ability to simultaneously serve as either an activator or a repressor of epithelial or mesenchymal gene expression, respectively. Keywords: EMT, H2A.Z, nucleosomes, TGF-β, gene regulation, promoter organization, cell plasticity, differentiation, chromati

The Histone Variant H2A.Z Is a Master Regulator of the Epithelial-Mesenchymal Transition

Epithelial-mesenchymal transition (EMT) is a profound example of cell plasticity that is crucial for embryonic development and cancer. Although it has long been suspected that chromatin-based mechanisms play a role in this process, no master regulator that can specifically regulate EMT has been identified to date. Here, we show that H2A.Z can coordinate EMT by serving as either an activator or repressor of epithelial or mesenchymal gene expression, respectively. Following induction of EMT by TGF-β, we observed an unexpected loss of H2A.Z across both downregulated epithelial and upregulated mesenchymal promoters. Strikingly, the repression of epithelial gene expression was associated with reduction of H2A.Z upstream of the transcription start site (TSS), while the activation of mesenchymal gene expression was dependent on removal of H2A.Z downstream of the TSS. Therefore, the ability of H2A.Z to regulate EMT is dependent on its position, either upstream or downstream of the TSS

Comparative epigenomics: an emerging field with breakthrough potential to understand evolution of epigenetic regulation

Epigenetic mechanisms regulate gene expression, thereby mediating the interaction between environment, genotype and phenotype. Changes to epigenetic regulation of genes may be heritable, permitting rapid adaptation of a species to environmental cues. However, most of the current understanding of epigenetic gene regulation has been gained from studies of mice and humans, with only a limited understanding of the conservation of epigenetic mechanisms across divergent taxa. The relative ease at which genome sequence data is now obtained and the advancements made in epigenomics techniques for non-model species provides a basis for carrying out comparative epigenomic studies across a wider range of species, making it possible to start unraveling the evolution of epigenetic mechanisms. We review the current knowledge of epigenetic mechanisms obtained from studying model organisms, give an example of how comparative epigenomics using non-model species is helping to trace the evolutionary history of X chromosome inactivation in mammals and explore the opportunities to study comparative epigenomics in biological systems displaying adaptation between species, such as the immune system and sex determination

Advanced Fashion Technology and Operations Management

Advanced Fashion Technology and Operations Management is a pivotal reference source for the latest development management strategies, fashion marketing, international business, and fashion entrepreneurship. Featuring extensive coverage across a range of relevant perspectives and topics, such as online shopping behavior, digital fashion, and e-commerce, this book is ideally designed for professionals, entrepreneurs, students, and researchers