• Screening for Mental Illness using GMHAT App of patients with Type 2 Diabetes Mellitus at a tertiary care hospital in India – A cross sectional study

      Jones, Steven; Majgi, Sumanth M.; Krishna, M.; Ebuenyi, I.; Mal, N. M.; University of Chester; FRAME - Mysore, India
      People with diabetes will have some coexisting mental illness most of the time and its evaluation and management are essential for the well‑being of the person. With this background, the Global Mental Health Assessment Tool (GMHAT) app was used to screen for mental illness among type 2 diabetes mellitus patients in Mysore, India. Methods: A cross‑sectional study was conducted on 100 patients who were on treatment for type 2 diabetes for at least 6 months. Interviews were conducted using a structured GMHAT. Results: The mean age of the study participants was 48.8 ± 11.6 years. Among the 100 diabetic patients, 39% (n = 39) had mental illness and among the 39 subjects, 21% had depression, 14% had anxiety disorder and 4% suffered from organic disorder. Apart from diabetes, 29, 17, 13, 27, 23 and 20% had hypertension, chronic kidney disease, liver disease, retinopathy, neuropathy and nephropathy, respectively, as comorbid conditions. Conclusion: There is a higher prevalence of mental illness in patients with diabetes mellitus. The prevalence rate increases with an increase in the chronicity of diabetes. The use of the GMHAT app will help in rapid assessment and accurate diagnoses
    • Second generation tyrosine kinase inhibitors prevent disease progression in high-risk (high CIP2A) chronic myeloid leukaemia patients.

      Lucas, Claire; Harris, Robert; Holcroft, Alison; Scott, Laura; Carmell, Natasha; McDonald, Elizabeth; Polydoros, Fotis; Clark, Richard (Nature, 2015-03-13)
      High cancerous inhibitor of PP2A (CIP2A) protein levels at diagnosis of chronic myeloid leukaemia (CML) are predictive of disease progression in imatinib-treated patients. It is not known whether this is true in patients treated with second generation tyrosine kinase inhibitors (2G TKI) from diagnosis, and whether 2G TKIs modulate the CIP2A pathway. Here, we show that patients with high diagnostic CIP2A levels who receive a 2G TKI do not progress, unlike those treated with imatinib (P=<0.0001). 2G TKIs induce more potent suppression of CIP2A and c-Myc than imatinib. The transcription factor E2F1 is elevated in high CIP2A patients and following 1 month of in vivo treatment 2G TKIs suppress E2F1 and reduce CIP2A; these effects are not seen with imatinib. Silencing of CIP2A, c-Myc or E2F1 in K562 cells or CML CD34+ cells reactivates PP2A leading to BCR-ABL suppression. CIP2A increases proliferation and this is only reduced by 2G TKIs. Patients with high CIP2A levels should be offered 2G TKI treatment in preference to imatinib. 2G TKIs disrupt the CIP2A/c-Myc/E2F1 positive feedback loop, leading to lower disease progression risk. The data supports the view that CIP2A inhibits PP2Ac, stabilising E2F1, creating a CIP2A/c-Myc/E2F1 positive feedback loop, which imatinib cannot overcome.
    • The sedentary office: an expert statement on the growing case for change towards better health and productivity

      Buckley, John P.; Hedge, Alan; Yates, Thomas; Copeland, Robert J.; Loosemore, Michael; Hamer, Mark; Bradley, Gavin; Dunstan, David W.; University Centre Shrewsbury (BMJ, 2015-06-01)
      An international group of experts convened to provide guidance for employers to promote the avoidance of prolonged periods of sedentary work. The set of recommendations was developed from the totality of the current evidence, including long-term epidemiological studies and interventional studies of getting workers to stand and/or move more frequently. The evidence was ranked in quality using the four levels of the American College of Sports Medicine. The derived guidance is as follows: for those occupations which are predominantly desk based, workers should aim to initially progress towards accumulating 2 h/day of standing and light activity (light walking) during working hours, eventually progressing to a total accumulation of 4 h/day (prorated to part-time hours). To achieve this, seated-based work should be regularly broken up with standing-based work, the use of sit–stand desks, or the taking of short active standing breaks. Along with other health promotion goals (improved nutrition, reducing alcohol, smoking and stress), companies should also promote among their staff that prolonged sitting, aggregated from work and in leisure time, may significantly and independently increase the risk of cardiometabolic diseases and premature mortality. It is appreciated that these recommendations should be interpreted in relation to the evidence from which they were derived, largely observational and retrospective studies, or short-term interventional studies showing acute cardiometabolic changes. While longer term intervention studies are required, the level of consistent evidence accumulated to date, and the public health context of rising chronic diseases, suggest initial guidelines are justified. We hope these guidelines stimulate future research, and that greater precision will be possible within future iterations.
    • A single dose of ChAdOx1 Chik vaccine induces neutralising antibodies against four chikungunya virus lineages in a phase 1 clinical trial

      Folegatti, Pedro M.; Harrison, Kate; Preciado-Llanes, Lorena; Ramos Lopez, Fernando; Bittaye, Mustapha; Kim, Young Chan; Flaxman, Amy; Bellamy, Duncan; Makinson, Rebecca; Sheridan, Jonathan; et al. (Nature Research, 2021-07-30)
      Chikungunya virus (CHIKV) is a reemerging mosquito-borne virus that causes swift outbreaks. Major concerns are the persistent and disabling polyarthralgia in infected individuals. Here we present the results from a first-in-human trial of the candidate simian adenovirus vectored vaccine ChAdOx1 Chik, expressing the CHIKV full-length structural polyprotein (Capsid, E3, E2, 6k and E1). 24 adult healthy volunteers aged 18–50 years, were recruited in a dose escalation, open-label, nonrandomized and uncontrolled phase 1 trial (registry NCT03590392). Participants received a single intramuscular injection of ChAdOx1 Chik at one of the three preestablished dosages and were followed-up for 6 months. The primary objective was to assess safety and tolerability of ChAdOx1 Chik. The secondary objective was to assess the humoral and cellular immunogenicity. ChAdOx1 Chik was safe at all doses tested with no serious adverse reactions reported. The vast majority of solicited adverse events were mild or moderate, and self-limiting in nature. A single dose induced IgG and Tcell responses against the CHIKV structural antigens. Broadly neutralizing antibodies against the four CHIKV lineages were found in all participants and as early as 2 weeks after vaccination. In summary, ChAdOx1 Chik showed excellent safety, tolerability and 100% PRNT50 seroconversion after a single dose.
    • Small-RNA Sequencing Reveals Altered Skeletal Muscle microRNAs and snoRNAs Signatures in Weanling Male Offspring from Mouse Dams Fed a Low Protein Diet during Lactation

      Kanakis, Ioannis; Alameddine, Moussira; Folkes, Leighton; Moxon, Simon; Myrtziou, Ioanna; Ozanne, Susan E.; Peffers, Mandy J.; Goljanek-Whysall, Katarzyna; Vasilaki, Aphrodite; University of Liverpool; University of Chester; University of East Anglia; University of Cambridge; NUI Galway (MDPI, 2021-05-11)
      Maternal diet during gestation and lactation affects the development of skeletal muscles in offspring and determines muscle health in later life. In this paper, we describe the association between maternal low protein diet-induced changes in offspring skeletal muscle and the differential expression (DE) of small non-coding RNAs (sncRNAs). We used a mouse model of maternal protein restriction, where dams were fed either a normal (N, 20%) or a low protein (L, 8%) diet during gestation and newborns were cross-fostered to N or L lactating dams, resulting in the generation of NN, NL and LN offspring groups. Total body and tibialis anterior (TA) weights were decreased in weanling NL male offspring but were not different in the LN group, as compared to NN. However, histological evaluation of TA muscle revealed reduced muscle fibre size in both groups at weaning. Small RNA-sequencing demonstrated DE of multiple miRs, snoRNAs and snRNAs. Bioinformatic analyses of miRs-15a, -34a, -122 and -199a, in combination with known myomiRs, confirmed their implication in key muscle-specific biological processes. This is the first comprehensive report for the DE of sncRNAs in nutrition-associated programming of skeletal muscle development, highlighting the need for further research to unravel the detailed molecular mechanisms.
    • Social and Clinical Correlates of Stimulant Use Disorder (Mephedrone) in a Tertiary Mental Health Setting in Mumbai: A Pilot Exploratory Study

      Rao, S. Poornima; Kale, Vinayak Pandurang; Panigrahi, Sunilkumar; Krishna, Murali; Jones, Steven; Majgi, Sumanth Mallikarjuna; Bharath, D. U.; University of Chester
      Introduction: Increasing mephedrone use is a major public health concern in India. There are limited data on sociodemographic determinants and psychiatric comorbidity associated with stimulant use disorder (mephedrone) (SUD‑M) from India. Aim: The primary objective of this study was to report the clinical and social correlates of SUD‑M among those presenting to specialist mental health services in Mumbai, India. Methods: Patients with SUD-M were recruited from a clinical setting. Standardized culturally validated assessments were carried out to obtain information about sociodemographics and mental health: comorbid psychopathology Brief Psychiatric Rating Scale, Hamilton Anxiety Rating Scale, Hamilton Depression Rating Scale, and Minnesota Multiphasic Personality Inventory‑2 for personality traits and a clinical assessment for diagnoses of mental disorders. Results: Seventy patients (aged between 21 and 30 years, of whom 58 men) with SUD-M consented. SUD‑M was more common among young men from the low socioeconomic position. The most common reasons for choosing mephedrone over other substances were better high from the drug and peer pressure. There were no associations between sociodemographic factors with the severity of SUD-M. Around 40% of the patients with SUD-M had psychiatric comorbidity. Psychotic disorders and anxiety symptoms were most common. Family history of substance use, comorbid substance use, and comorbid psychiatric disorders were directly related to the severity of SUD-M. Conclusions: This was a cross‑sectional study with a relatively smaller sample size of self‑nominating participantslimiting the generalizability of findings to a wider population. Therapeutic implication of this finding is that prompt attention and treatment of the comorbid psychiatric disorder is essential while treating patients with SUD-M. Further population-based studies are recommended for a better understanding of the burden of SUD-M.
    • Spinal motor neurite outgrowth over glial scar inhibitors is enhanced by coculture with bone marrow stromal cells

      Wright, Karina; Johnson, William Eustace Basil; Uchida, Kenzo; Bara, Jennifer J.; Roberts, Sally; Masari, Wagih E.; Aston University; Keele University
      BACKGROUND CONTEXT: Transplantation of bone marrow cells into spinal cord lesions promotes functional recovery in animal models, and recent clinical trials suggest possible recovery also in humans. The mechanisms responsible for these improvements are still unclear. PURPOSE: To characterize spinal cord motor neurite interactions with human bone marrow stromal cells (MSCs) in an in vitro model of spinal cord injury (SCI). STUDY DESIGN/SETTING: Previously, we have reported that human MSCs promote the growth of extending sensory neurites from dorsal root ganglia (DRG), in the presence of some of the molecules present in the glial scar, which are attributed with inhibiting axonal regeneration after SCI. We have adapted and optimized this system replacing the DRG with a spinal cord culture to produce a central nervous system (CNS) model, which is more relevant to the SCI situation. METHODS: We have developed and characterized a novel spinal cord culture system. Human MSCs were cocultured with spinal motor neurites in substrate choice assays containing glial scar–associated inhibitors of nerve growth. In separate experiments, MSC-conditioned media were analyzed and added to spinal motor neurites in substrate choice assays. RESULTS: As has been reported previously with DRG, substrate-bound neurocan and Nogo-A repelled spinal neuronal adhesion and neurite outgrowth, but these inhibitory effects were abrogated in MSC/spinal cord cocultures. However, unlike DRG, spinal neuronal bodies and neurites showed no inhibition to substrates of myelin-associated glycoprotein. In addition, the MSC secretome contained numerous neurotrophic factors that stimulated spinal neurite outgrowth, but these were not sufficient stimuli to promote spinal neurite extension over inhibitory concentrations of neurocan or Nogo-A. CONCLUSIONS: These findings provide novel insight into how MSC transplantation may promote regeneration and functional recovery in animal models of SCI and in the clinic, especially in the chronic situation in which glial scars (and associated neural inhibitors) are well established. In addition, we have confirmed that this CNS model predominantly comprises motor neurons via immunocytochemical characterization. We hope that this model may be used in future research to test various other potential interventions for spinal injury or disease states
    • SS-31 attenuates TNF-α induced cytokine release from C2C12 myotubes

      Nye, Gareth; Lightfoot, Adam; Sakellariou, Giorgos; McArdle, Francis; Jackson, Malcolm; Griffiths, Richard; McArdle, Anne; University of Liverpool (Elsevier, 2015-08-10)
      TNF-α is a key inflammatory mediator and is proposed to induce transcriptional responses via the mitochondrial generation of Reactive Oxygen Species (ROS). The aim of this study was to determine the effect of TNF-α on the production of myokines by skeletal muscle. Significant increases were seen in the release of IL-6, MCP-1/CCL2, RANTES/CCL5 and KC/CXCL1 and this release was inhibited by treatment with Brefeldin A, suggesting a golgi-mediated release of cytokines by muscle cells. An increase was also seen in superoxide in response to treatment with TNF-α, which was localised to the mitochondria and this was also associated with activation of NF-κB. The changes in superoxide, activation of NF-kB and release of myokines were attenuated following pre-treatment with SS-31 peptide indicating that the ability of TNF-α to induce myokine release may be mediated through mitochondrial superoxide, which is, at least in part, associated with activation of the redox sensitive transcription factor NF-kB.
    • Standards and core components for cardiovascular disease prevention and rehabilitation; BACPR

      Cowie, Aynsley; Buckley, John P.; Doherty, Patrick; Furze, Gill; Hayward, Jo; Jones, Jennifer; Speck, Linda; Dalal, Hayes; Mills, Joseph; University Centre Shrewsbury (BMJ, 2019-01-30)
      In 2017, the British Association for Cardiovascular Prevention and Rehabilitation published its official document detailing standards and core components for cardiovascular prevention and rehabilitation. Building on the success of previous editions of this document (published in 2007 and 2012), the 2017 update aims to further emphasise to commissioners, clinicians, politicians and the public the importance of robust, quality indicators of cardiac rehabilitation (CR) service delivery. Otherwise, its overall aim remains consistent with the previous publications—to provide a precedent on which all effective cardiovascular prevention and rehabilitation programmes are based and a framework for use in assessment of variation in service delivery quality. In this 2017 edition, the previously described seven standards and core components have both been revised to six, with a greater focus on measurable clinical outcomes, audit and certification. The principles within the updated document underpin the six-stage pathway of care for CR, and reflect the extensive evidence base now available within the field. To help improve current services, close collaboration between commissioners and CR providers is advocated, with use of the CR costing tool in financial planning of programmes. The document specifies how quality assurance can be facilitated through local audit, and advocates routine upload of individual-level data to the annual British Heart Foundation National Audit of Cardiac Rehabilitation, and application for national certification ensuring attainment of a minimum quality standard. Although developed for the UK, these standards and core components may be applicable to other countries.
    • Stent Frame Movement Following Endovascular Aneurysm Sealing in the Abdominal Aorta

      Yafawi, Asma; McWilliams, Richard G.; Fisher, Robert K.; England, Andrew; Karouki, Maria; Torella, Francesco (SAGE Publications, 2018-11-28)
    • Stent Migration Following Endovascular Sealing of Abdominal Aortic Aneurysms

      Yafawi, Asma; McWilliams, Richard G.; Fisher, Robert K.; England, Andrew; Karouki, Maria; Torella, Francesco (Elsevier, 2019-12-09)
    • Submaximal Exercise Testing in Cardiovascular Rehabilitation Settings (BEST Study)

      Reed, Jennifer L; Cotie, LM; Cole, CA; Harris, Jennifer; Moran, B; Scott, K; Buckley, John P.; Pipe, Andrew L; University of Ottawa, University Centre Shrewsbury/Chester
      Abstract BACKGROUND: This study compared changes in measured versus predicted peak aerobic power (V̇O2 peak) following cardiovascular rehabilitation (CR). Peak cardiopulmonary exercise testing (CPET) results were compared to four V̇O2 peak estimation methods: the submaximal modified Bruce treadmill, Astrand-Ryhming cycle ergometer, and Chester step tests, and the Duke Activity Status Index (DASI). METHODS: Adults with cardiovascular disease (CVD) who completed a 12-week CR program were assessed at baseline and 12 weeks follow-up. CPET, the DASI and three subsequent submaximal exercise tests were performed in a random order. RESULTS: Of the 50 adults (age: 57 ± 11 years) who participated, 46 completed the 12-week CR program and exercise tests. At baseline 69, 68, and 38% of the treadmill, step and cycle tests were successfully completed, respectively. At follow-up 67, 80, and 46% of the treadmill, step and cycle tests were successfully completed, respectively. No severe adverse events occurred. Significant improvements in V̇O2 peak were observed with CPET (3.6 ± 5.5 mL.kg-1.min-1, p < 0.001) and the DASI (2.3 ± 4.2 mL.kg-1.min-1, p < 0.001). Bland-Altman plots of the change in V̇O2 peak between CPET and the four V̇O2 peak estimation methods revealed the following: a proportional bias and heteroscedastic 95% limits of agreement (95% LoA) for the treadmill test, and for the cycle and step tests and DASI, mean bias' and 95% LoA of 1.0 mL.kg-1.min-1 (21.3, -19.3), 1.4 mL.kg-1.min-1 (15.0, -12.3) and 1.0 mL.kg-1.min-1 (13.8, -11.8), respectively. CONCLUSION: Given the greater number of successful tests, no serious adverse events and acceptable mean bias, the step test appears to be a valid and safe method for assessing group-level mean changes in V̇O2 peak among patients in CR. The DASI also appears to be a valid and practical questionnaire. Wide limits of agreement, however, limit their use to predict individual-level changes. Copyright © 2020 Reed, Cotie, Cole, Harris, Moran, Scott, Terada, Buckley and Pipe. KEYWORDS: cardiovascular diseases; exercise test; physiology; rehabilitation; submaximal
    • Systematic review and meta-analysis of group cognitive behavioural psychotherapy for sub-clinical depression

      Krishna, Murali; Lepping, Peter; Jones, Steven; Lane, Stephen; University of Chester (Elsevier, 2015-06-04)
      Key Points • Group CBT for patients with sub-threshold depression has a significant effect on depressive symptomatology at post treatment in both working age and older adult population. • Group CBT does not appear to reduce the incidence of major depressive disorders. • Group CBT has minimal or no effect on depressive symptomatology during follow-up. The article considers group psychotherapy in sub-threshold depression to investigate if group psychological interventions reduce depressive symptoms post treatment, and whether these interventions result in a reduced incidence of new cases of major depressive disorder.
    • Targeting long non-coding RNAs (lncRNAs) with oligonucleotides in cancer therapy

      Pickard, Mark R.; Williams, Gwyn T.; University of Chester; Keele University (AME Publishing Company, 2016-10)
      No abstract
    • The Comparative Effects of Mesenchymal Stem Cell Transplantation Therapy for Spinal Cord Injury in Humans and Animal Models: A Systematic Review and Meta-Analysis

      Johnson, Louis D. V.; email: louisdvj@gmail.com; Pickard, Mark R.; email: m.pickard@chester.ac.uk; Johnson, William E. B.; email: eustace.johnson@chester.ac.uk (MDPI, 2021-03-16)
      Animal models have been used in preclinical research to examine potential new treatments for spinal cord injury (SCI), including mesenchymal stem cell (MSC) transplantation. MSC transplants have been studied in early human trials. Whether the animal models represent the human studies is unclear. This systematic review and meta-analysis has examined the effects of MSC transplants in human and animal studies. Following searches of PubMed, Clinical Trials and the Cochrane Library, published papers were screened, and data were extracted and analysed. MSC transplantation was associated with significantly improved motor and sensory function in humans, and significantly increased locomotor function in animals. However, there are discrepancies between the studies of human participants and animal models, including timing of MSC transplant post-injury and source of MSCs. Additionally, difficulty in the comparison of functional outcome measures across species limits the predictive nature of the animal research. These findings have been summarised, and recommendations for further research are discussed to better enable the translation of animal models to MSC-based human clinical therapy.
    • The Differences in Acute Management of Asthma in Adults and Children

      Chavasse, Richard; Scott, Stephen (Frontiers Media S.A., 2019-03-11)
      Acute asthma or wheeze is a common presentation to emergency services for both adults and children. Although there are phenotypic differences between asthma syndromes, the management of acute symptoms follow similar lines. This article looks at the similarities and differences in approaches for children and adults. Some of these may be age dependent, such as the physiological parameters used to define the severity of the attack or the use of age appropriate inhaler devices. Other differences may reflect the availability of evidence. In other areas there is conflicting evidence between adult and pediatric studies such as a temporary increase in dose of inhaled corticosteroids during an acute attack. Overall there are more similarities than differences.
    • Training nurses in Mental Health Assessment using GMHAT/PC in India

      Jones, Steven; Keenan, Paul; Danivas, Vijay; Krishna, Murali; Rajendra, Rajgopal; University of Chester (Indian Psychiatric Society, 2017-07-23)
      Book chapter; Training of Indian general nurses in a hospital setting required the structure offered by the Global Mental Health Assessment Tool (GMHAT) that would provide a framework to underpin mental health assessment training. Attitudes of those undertaking the training and current levels of knowledge and awareness to mental health issues was explored prior to any training occurring in the use of GMHAT, that we considered fundamental to good mental health practice
    • Treatment-Free Remission in Chronic Myeloid Leukemia: Can We Identify Prognostic Factors?

      Lucas, Claire; Saifullah, Hilbeen H.; University of Chester; University of Liverpool (MDPI, 2021-08-19)
      Following the development of tyrosine kinase inhibitors (TKI), the survival of patients with chronic myeloid leukaemia (CML) drastically improved. With the introduction of these agents, CML is now considered a chronic disease for some patients. Taking into consideration the side effects, toxicity, and high cost, discontinuing TKI became a goal for patients with chronic phase CML. Patients who achieved deep molecular response (DMR) and discontinued TKI, remained in treatment-free remission (TFR). Currently, the data from the published literature demonstrate that 40–60% of patients achieve TFR, with relapses occurring within the first six months. In addition, almost all patients who relapsed regained a molecular response upon retreatment, indicating TKI discontinuation is safe. However, there is still a gap in understanding the mechanisms behind TFR, and whether there are prognostic factors that can predict the best candidates who qualify for TKI discontinuation with a view to keeping them in TFR. Furthermore, the information about a second TFR attempt and the role of gradual de-escalation of TKI before complete cessation is limited. This review highlights the factors predicting success or failure of TFR. In addition, it examines the feasibility of a second TFR attempt after the failure of the first one, and the current guidelines concerning TFR in clinical practice.
    • Tuberculosis notification in a private tertiary care teaching hospital in South India: a mixed-methods study

      Siddaiah, Archana; Ahmed, Mohammad Naseer; Kumar, Ajay M. V.; D’Souza, George; Wilkinson, Ewan; Maung, Thae Maung; Rodrigues, Rashmi (BMJ Publishing Group, 2019-02-05)
      Objectives India contributes approximately 25% of the ‘missing’ cases of tuberculosis (TB) globally. Even though ~50% of patients with TB are diagnosed and treated within India’s private sector, few are notified to the public healthcare system. India’s TB notification policy mandates that all patients with TB are notified through Nikshay (TB notification portal). We undertook this study in a private hospital to assess the proportion notified and factors affecting TB notifications. We explored barriers and probable solutions to TB notification qualitatively from health provider’s perspective. Study setting Private, tertiary care, teaching hospital in Bengaluru, South India. Methodology This was a mixed-methods study. Quantitative component comprised a retrospective review of hospital records between 1 January 2015 and 31 December 2017 to determine TB notifications. The qualitative component comprised key informant interviews and focus groups to elicit the barriers and facilitators of TB notification. Results Of 3820 patients diagnosed and treated, 885 (23.2%) were notified. Notifications of sputum smear-positive patients were significantly more likely, while notifications of children were less likely. Qualitative analysis yielded themes reflecting the barriers to TB notification and their solutions. Themes related to barriers were: (1) basic diagnostic procedures and treatment promote notification; (2) misconceptions regarding notification and its process are common among healthcare providers; (3) despite a national notification system other factors have prevented notification of all patients; and (4) establishing hospital systems for notification will go a long way in improving notifications. Conclusions The proportion of patients with TB notified by the hospital was low. A comprehensive approach both by the hospital management and the national TB programme is necessary for improving notification. This includes improving awareness among healthcare providers about the requirement for TB notifications, establishing a single notification portal in hospital, digitally linking hospital records to Nikshay and designating one person to be responsible for notification.
    • Two independent proteomic approaches provide a comprehensive analysis of the synovial fluid proteome response to Autologous Chondrocyte Implantation

      Hulme, Charlotte H.; Wilson, Emma L.; Fuller, Heidi R.; Roberts, Sally; Richardson, James B.; Gallacher, Pete; Peffers, Mandy J.; Shirran, Sally L.; Botting, Catherine H.; Wright, Karina T.; et al. (BioMed Central, 2018-05-02)
      Background: Autologous chondrocyte implantation (ACI) has a failure rate of approximately 20%, but it is yet to be fully understood why. Biomarkers are needed that can pre-operatively predict in which patients it is likely to fail, so that alternative or individualised therapies can be offered. We previously used label-free quantitation (LF) with a dynamic range compression proteomic approach to assess the synovial fluid (SF) of ACI responders and non-responders. However, we were able to identify only a few differentially abundant proteins at baseline. In the present study, we built upon these previous findings by assessing higher-abundance proteins within this SF, providing a more global proteomic analysis on the basis of which more of the biology underlying ACI success or failure can be understood. Methods: Isobaric tagging for relative and absolute quantitation (iTRAQ) proteomic analysis was used to assess SF from ACI responders (mean Lysholm improvement of 33; n = 14) and non-responders (mean Lysholm decrease of 14; n = 13) at the two stages of surgery (cartilage harvest and chondrocyte implantation). Differentially abundant proteins in iTRAQ and combined iTRAQ and LF datasets were investigated using pathway and network analyses. Results: iTRAQ proteomic analysis confirmed our previous finding that there is a marked proteomic shift in response to cartilage harvest (70 and 54 proteins demonstrating ≥ 2.0-fold change and p < 0.05 between stages I and II in responders and non-responders, respectively). Further, it highlighted 28 proteins that were differentially abundant between responders and non-responders to ACI, which were not found in the LF study, 16 of which were altered at baseline. The differential expression of two proteins (complement C1s subcomponent and matrix metalloproteinase 3) was confirmed biochemically. Combination of the iTRAQ and LF proteomic datasets generated in-depth SF proteome information that was used to generate interactome networks representing ACI success or failure. Functional pathways that are dysregulated in ACI non-responders were identified, including acute-phase response signalling. Conclusions: Several candidate biomarkers for baseline prediction of ACI outcome were identified. A holistic overview of the SF proteome in responders and non-responders to ACI  has been profiled, providing a better understanding of the biological pathways underlying clinical outcome, particularly the differential response to cartilage harvest in non-responders.