Comparison of different methods for preparation and characterization of total RNA from cartilage samples to uncover osteoarthritis in vivo

<p>Abstract</p> <p>Background</p> <p>The isolation of intact RNA can be very difficult when tissues are used that contain many RNAses or that are hard to homogenize, e.g. cartilage samples. Additionally, cartilaginous tissues are characterized by a low cellularity and an abundance of extracellular matrix (ECM) molecules. But given the growing interest in understanding pathogenesis of degenerative diseases, e.g. osteoarthritis (OA) and rheumatoid arthritis (RA), studies have to consider expression pattern of cells in its natural environment.</p> <p>Findings</p> <p>We compared the current RNA isolation methods for the extraction of high-quality RNA of snap-frozen biopsies from limited amounts of hypocellular cartilaginous tissue. The focus of the study was to gather information about procedure-related differences in RNA quality and yield. Here, we describe two protocols, the phenol/chloroform-free filter-based method (RN<it>Aqueous</it>™ kit) and the combined protocol (TRIzol^®/RNeasy Mini™ kit), working in a reproducible and reliable manner.</p> <p>Conclusions</p> <p>We conclude that preparation, storage, homogenization, and quality control are altogether critical steps for in-depth analysis of differential gene expression, especially in hypocellular tissues with highly crosslinked ECM like cartilage.</p

Development of a DNA-based microarray for the detection of zoonotic pathogens in rodent species

The demand for diagnostic tools that allow simultaneous screening of samples for multiple pathogens is increasing because they overcome the limitations of other methods, which can only screen for a single or a few pathogens at a time. Microarrays offer the advantages of being capable to test a large number of samples simultaneously, screening for multiple pathogen types per sample and having comparable sensitivity to existing methods such as PCR. Array design is often considered the most important process in any microarray experiment and can be the deciding factor in the success of a study. There are currently no microarrays for simultaneous detection of rodent-borne pathogens. The aim of this report is to explicate the design, development and evaluation of a microarray platform for use as a screening tool that combines ease of use and rapid identification of a number of rodent-borne pathogens of zoonotic importance. Nucleic acid was amplified by multiplex biotinylation PCR prior to hybridisation onto microarrays. The array sensitivity was comparable to standard PCR, though less sensitive than real-time PCR. The array presented here is a prototype microarray identification system for zoonotic pathogens that can infect rodent species

Attenuated cardiovascular reactivity is related to higher anxiety and fatigue symptoms in truck drivers

ACKNOWLEDGEMENTS The authors would like to thank all the truck drivers who participated in this study. The data presented in this paper were collected as part of the baseline measures from the “Structured Health Intervention For Truckers (SHIFT)” randomized controlled trial, which is funded by the NIHR Public Health Research Programme (reference: NIHR PHR 15/190/42). SAC, JAK, AS and NJP are supported by the NIHR Leicester Biomedical Research Centre—Lifestyle theme. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. The first author (AG) has received funding for their PhD Studentship from the Colt Foundation (reference: JD/618). The Colt Foundation had no role in study design; election, synthesis, and interpretation of data; writing of the report; or the decision to submit the manuscript for publicationPeer reviewedPublisher PD

Time in Nature Associated with Decreased Fatigue in UK Truck Drivers

Funding: The data presented in this paper were collected as part of the ‘Structured Health Interven- tion For Truckers (SHIFT)’ randomised controlled trial. This research was funded by the National Institute for Health Research (NIHR) Public Health Research Programme (reference: NIHR PHR 15/190/42). Funding Acquisition, S.A.C., J.A.K., V.V-M. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. Acknowledgments: SAC: JAK, AS and NJP are supported by the NIHR Leicester Biomedical Re- search Centre—Lifestyle theme. AG has received funding for their PhD Studentship from the Colt Foundation (reference: JD/618).Peer reviewedPublisher PD

The structured health intervention for truckers (SHIFT) cluster randomised controlled trial : a mixed methods process evaluation

Funding This project was funded by the National Institute for Health Research (NIHR) Public Health Research programme (reference: NIHR PHR 15/190/42). The study was also supported by the NIHR Leicester Biomedical Research Centre which is a partnership between University Hospitals of Leicester NHS Trust, Loughborough University, and the University of Leicester. Laura Gray is supported by the National Institute for Health Research (NIHR) Applied Research Collaboration East Midlands (ARC EM). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. Funding to cover intervention costs (Fitbits, cab workout equipment) was provided by the Higher Education Innovation Fund, via the Loughborough University Enterprise Projects Group. The Colt Foundation provided funding for a PhD Studentship, awarded to Amber Guest (reference: JD/618), which covered Amber’s time and contributions to this project. None of the funding bodies had any role in study design; election, synthesis, and interpretation of data; writing of the report; or the decision to submit the manuscript for publication. Acknowledgements We gratefully acknowledge the support provided by senior Health and Safety personnel and Transport Managers at our partner logistics company in facilitating this research. We also thank all participants for taking part. We are grateful to the independent members of the Trial Steering Committee for their continued support and advice throughout the trial: Dr. Derrick Bennett, Prof Emma McIntosh, Prof Petra Wark and Mr. Paul Gardiner.Peer reviewedPublisher PD

Drivers with and without Obesity Respond Differently to a Multi-Component Health Intervention in Heavy Goods Vehicle Drivers

Funding This project was funded by the National Institute for Health Research (NIHR) Public Health Research programme (reference: NIHR PHR 15/190/42). The study was also supported by the NIHR Leicester Biomedical Research Centre which is a partnership between University Hospitals of Leicester NHS Trust, Loughborough University and the University of Leicester. Laura Gray is supported by the National Institute for Health Research (NIHR) Applied Research Collaboration East Midlands (ARC EM). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. Funding to cover the intervention costs (Fitbits and cab workout equipment) was provided by the Higher Education Innovation Fund, via the Loughborough University Enterprise Projects Group. The Colt Foundation provided funding for a PhD Studentship, awarded to Amber Guest (reference: JD/618), which covered Amber’s time and contributions to this project. The funders played no role in study design, data collection, data analysis, data interpretation or in the preparation of this manuscript.Peer reviewedPublisher PD

A study of tuberculosis in road traffic-killed badgers on the edge of the British bovine TB epidemic area

The role of badgers in the geographic expansion of the bovine tuberculosis (bTB) epidemic in England is unknown: indeed there have been few published studies of bTB in badgers outside of the Southwest of England where the infection is now endemic in cattle. Cheshire is now on the edge of the expanding area of England in which bTB is considered endemic in cattle. Previous studies, over a decade ago when bovine infection was rare in Cheshire, found no or only few infected badgers in the south eastern area of the county. In this study, carried out in 2014, road-killed badgers were collected through a network of local stakeholders (farmers, veterinarians, wildlife groups, government agencies), and Mycobacterium bovis was isolated from 21% (20/94) badger carcasses. Furthermore, there was strong evidence for co-localisation of M. bovis SB0129 (genotype 25) infection in both badgers and cattle herds at a county scale. While these findings suggest that both badgers and cattle are part of the same geographically expanding epidemic, the direction of any cross-species transmission and the drivers of this expansion cannot be determined. The study also demonstrated the utility of using road-killed badgers collected by stakeholders as a means of wildlife TB surveillance

Zoonotic Chlamydiaceae Species Associated with Trachoma, Nepal - Volume 19, Number 12—December 2013 - Emerging Infectious Diseases journal - CDC

Trachoma is the leading cause of preventable blindness. Commercial assays do not discriminate among all Chlamydiaceae species that might be involved in trachoma. We investigated whether a commercial Micro-ArrayTube could discriminate Chlamydiaceae species in DNA extracted directly from conjunctival samples from 101 trachoma patients in Nepal. To evaluate organism viability, we extracted RNA, reverse transcribed it, and subjected it to quantitative real-time PCR. We found that 71 (70.3%) villagers were infected. ArrayTube sensitivity was 91.7% and specificity was 100% compared with that of real-time PCR. Concordance between genotypes detected by microarray and ompA genotyping was 100%. Species distribution included 54 (76%) single infections with Chlamydia trachomatis, C. psittaci, C. suis, or C. pecorum, and 17 (24%) mixed infections that includied C. pneumoniae. Ocular infections were caused by 5 Chlamydiaceae species. Additional studies of trachoma pathogenesis involving Chlamydiaceae species other than C. trachomatis and their zoonotic origins are needed