Comparison of Canine Spermatozoa Motility, Morphology, and RNA Integrity Using Three Different Cell Purification Solutions

Transcriptome gene expression studies of sperm RNA have been utilized in a variety of different species to investigate causes of male infertility. Previous research investigators have optimized cell separation and RNA isolation techniques for each species of interest. To date, no study has been completed for dogs. The objective of this thesis research was to investigate the efficacy of various cell separation techniques in separating sperm cells from somatic cells in the ejaculate to yield a sample of total pure sperm RNA that could be used for a future downstream application. Comparisons were made between the conventional swim-up method and two commercial density gradient centrifugation (DGC) solutions (Bovipure[supercript]TM and Equipure[superscript]TM, Nidacon International, Mölndal, Sweden) Prior to and just following cell separation, total motility, normal morphology, and sperm count were determined for each method. Following cell separation, total RNA was isolated from each sample and RNA quantity and quality was determined via spectrophotometry and reverse transcriptase polymerase chain reaction. The presence of somatic cell RNA was used to determine purity of the cell separation method. The DGC methods were superior in separating sperm with higher percent normal morphology and higher total motility than the swim-up method. Additionally, the DGC methods were superior at producing a more pure RNA sample than samples not treated with any separation technique (control). This research shows that the DGC methods should be used to separate canine sperm cells prior to RNA isolation for sperm-specific transcriptome applications

Predicting response to physiotherapy treatment for musculoskeletal shoulder pain : A systematic review

© 2013 Chester et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Background: People suffering from musculoskeletal shoulder pain are frequently referred to physiotherapy. Physiotherapy generally involves a multimodal approach to management that may include; exercise, manual therapy and techniques to reduce pain. At present it is not possible to predict which patients will respond positively to physiotherapy treatment. The purpose of this systematic review was to identify which prognostic factors are associated with the outcome of physiotherapy in the management of musculoskeletal shoulder pain. Methods. A comprehensive search was undertaken of Ovid Medline, EMBASE, CINAHL and AMED (from inception to January 2013). Prospective studies of participants with shoulder pain receiving physiotherapy which investigated the association between baseline prognostic factors and change in pain and function over time were included. Study selection, data extraction and appraisal of study quality were undertaken by two independent assessors. Quality criteria were selected from previously published guidelines to form a checklist of 24 items. The study protocol was prospectively registered onto the International Prospective Register of Systematic Reviews. Results: A total of 5023 titles were retrieved and screened for eligibility, 154 articles were assessed as full text and 16 met the inclusion criteria: 11 cohort studies, 3 randomised controlled trials and 2 controlled trials. Results were presented for the 9 studies meeting 13 or more of the 24 quality criteria. Clinical and statistical heterogeneity resulted in qualitative synthesis rather than meta-analysis. Three studies demonstrated that high functional disability at baseline was associated with poor functional outcome (p ≤ 0.05). Four studies demonstrated a significant association (p ≤ 0.05) between longer duration of shoulder pain and poorer outcome. Three studies, demonstrated a significant association (p ≤ 0.05) between increasing age and poorer function; three studies demonstrated no association (p > 0.05). Conclusion: Associations between prognostic factors and outcome were often inconsistent between studies. This may be due to clinical heterogeneity or type II errors. Only two baseline prognostic factors demonstrated a consistent association with outcome in two or more studies; duration of shoulder pain and baseline function. Prior to developing a predictive model for the outcome of physiotherapy treatment for shoulder pain, a large adequately powered prospective cohort study is required in which a broad range of prognostic factors are incorporated.Peer reviewe