• Attitudes of Female Warders towards inmate who self-harm: A pilot Exploratory Study from an Inner-city Prison in South India

      Jones, Steven; Kumar, Pradeep; Eost-Telling, Charlotte; Kirshna, Murali; University of Chester (The Indian Society of Criminology; National Law University Delhi, 07-2020)
      Self-harm is a global public health challenge. The management and treatment of those who self-harm is emotionally challenging, and can sometimes manifest in negative attitudes amongst staff who provide care. Health professional’s attitudes towards deliberate self-harm have been studied globally, however, evidence regarding prison staff attitudes is sparse, and particularly lacking in India. The primary aim of this study was to explore the attitudes of female prison warders towards prison inmates who self-harm in an Indian setting. A cross-sectional survey using a questionnaire to measure knowledge and attitudes was administered to prison warders from one city prison in South India. Out of the 210 approached to participate, 170 female warders completed the survey questionnaire. In general, sociodemographic factors of the prison warders were unrelated to their attitudes towards self-harm, and a negligible few had received any training specific to self-harm. A series of educational and skills recommendations have been developed from the study, which can be used to inform intervention initiatives and further, provide a basis for cross-cultural professional comparison studies. Current resources, cultures, practice and context must be considered in any future interventions aimed at progressing the evidence base further. In addition, training and education for staff should include information on knowledge and attitudes about causes, reasons, motivations, forms and purpose of self-harm. Records of staff responses to those who self-harm, irrespective of setting, should include assessment, management, interventions undertaken and incorporated daily practice. Importantly, this work may influence prisoner treatment outcomes and is worthy of further study.
    • Physical health impairment, disability and suicidal intent among self-harm survivors in South India

      Jones, Steven; Somashekar, R; Bharath, D.U; Maiji, Sumanth M; Taylor, Lou; Nagaraj, Santhosh; Krishna, Murali; Mysore Medical College and Research Institute; University of Chester; CSI Holdsworth Memorial Hospital; FRAMe; Viveka Hospital
      Background: Suicide is major public health concern in India. There are limited data examining the relationship between health impairment, disability and severity of suicidal intent. The aim of the study was to examine the associations of health impairment and disability with severity of suicidal intent among survivors following an act of self-harm. Methods: A pilot exploratory study of 453 self-harm survivors from a specialist hospital in South India. Sociodemographics, physical health impairment, disability (WHO Disability Schedule-II), suicidal intent, (Pierce suicide intent scale) and mental disorders were studied. Results: Arthritis was the most common physical impairment among self-harm survivors followed by gastrointestinal, sensory impairment and difficulty with mobilization. Nearly 10% of participants had some degree of functional impairment, with 38% experiencing severe physical pain in the week prior to self-harm. Past history of depression treatment, age, education and occupation influenced positively PSIS scores. There were significant associations between suicidal intent and disability. Conclusions: Indian self-harm survivors indicated complex relationships between physical health, disability and suicidal intent. Understanding these associations may help to develop suicide prevention strategies. Our findings suggest a need for integrating a comprehensive of physical health assessment in self harm survivors.
    • Comment on PP2A inhibition sensitizes cancer stem cells to ABL tyrosine kinase inhibitors in BCR-ABL human leukemia

      Perrotti, D; Agarwal, A; Lucas, Claire; Narla, g; Nevanini, p; Odero, m; Ruvolo, p; Verrills, n; University of Maryland; Imperial College London; Oregon Health and Science University; University of Chester; University of Michigan; University of Southern California; University of Navarra; MD Anderson Cancer Center; University of Newcastle (AAAS, 2019-07-17)
      LB100 does not sensitize CML stem cells to tyrosine kinase inhibitor–induced apoptosis.
    • Discovery of a Novel CIP2A Variant (NOCIVA) with clinical relevance in predicting TKI resistance in myeloid leukemias

      Makela, Eleonora; Pavic, Karolina; Varila, Taru M; Salmenniemi, Urpu; Löyttyniemi, Eliisa; Nagelli, Srikar G; Ammunét, Tea; Kähäri, Veli-Matti; Clark, Richard E; Elo, Laura L; et al.
      Purpose: Cancerous inhibitor of PP2A (CIP2A) is an oncoprotein that inhibits the tumor suppressor PP2A-B56a. However, CIP2A mRNA variants remain uncharacterized. Here, we report the discovery of a CIP2Asplicing variant, NOCIVA (NOvel CIp2a VAriant). Experimental Design: Characterization of CIP2A variants was performed by both 3' and 5' rapid amplification of cDNA ends from cancer cells. The function of NOCIVA was assessed by structural and molecular biology approaches. Its clinical relevance was studied in an acute myeloid leukemia (AML) patient cohort and two independent chronic myeloid leukemia (CML) cohorts. Results: NOCIVA contains CIP2A exons 1-13 fused to 349 nucleotides from CIP2A intron 13. Intriguingly, the first 39 nucleotides of the NOCIVA-specific sequence are in the coding frame with exon 13 of CIP2A and code for a 13 amino acid peptide tail nonhomologous to any known human protein sequence. Therefore, NOCIVA translates to a unique human protein. NOCIVA retains the capacity to bind to B56a, but whereas CIP2A is predominantly a cytoplasmic protein, NOCIVA translocates to the nucleus. Indicative of prevalent alternative splicing from CIP2A to NOCIVA in myeloid malignancies, AML and CML patient samples overexpress NOCIVA but not CIP2A mRNA. In AML, a high NOCIVA/CIP2A mRNA expression ratio is a marker for adverse overall survival. In CML, high NOCIVA expression is associated with inferior event-free survival among imatinib-treated patients, but not among patients treated with dasatinib or nilotinib. Conclusions: We discovered novel variant of the oncoprotein CIP2A and its clinical relevance in predicting tyrosine kinase inhibitor therapy resistance in myeloid leukemias.
    • Acute glycaemic management before, during and after exercise for cardiac rehabilitation participants with diabetes mellitus; a joint statement of the British and Canadian Associations of Cardiovascular Prevention and Rehabilitation, the International Council for Cardiovascular Prevention and Rehabilitation and the British Association of Sport and Exercise Sciences

      Buckley, J.P.; Riddell, Michael; Mellor, Duane; Bracken, Richard; Ross, Marie-Kristelle; LaGerche, Andre; Poirier, Paul; University of Chester; University College London; York University, Toronto; LMC Healthcare; Aston University; Swansea University College of Engineering; Laval University; Baker Heart and Diabetes Institute; St Vincent's Hospital Melbourne; Institut universitaire de cardiologie et de pneumologie de Québec
      Type 1 (T1) and type 2 (T2) diabetes mellitus (DM) are significant precursors and comorbidities to cardiovascular disease and prevalence of both types is still rising globally. Currently,~25% of participants (and rising) attending cardiac rehabilitation in Europe, North America and Australia have been reported to have DM (>90% have T2DM). While there is some debate over whether improving glycaemic control in those with heart disease can independently improve future cardiovascular health-related outcomes, for the individual patient whose blood glucose is well controlled, it can aid the exercise programme in being more efficacious. Good glycaemic management not only helps to mitigate the risk of acute glycaemic events during exercising, it also aids in achieving the requisite physiological and psycho-social aims of the exercise component of cardiac rehabilitation (CR). These benefits are strongly associated with effective behaviour change, including increased enjoyment, adherence and self-efficacy. It is known that CR participants with DM have lower uptake and adherence rates compared with those without DM. This expert statement provides CR practitioners with nine recommendations aimed to aid in the participant’s improved blood glucose control before, during and after exercise so as to prevent the risk of glycaemic events that could mitigate their beneficial participation.
    • Combined bezafibrate, medroxyprogesterone acetate and valproic acid treatment inhibits osteosarcoma cell growth without adversely affecting normal mesenchymal stem cells.

      Sheard, Jonathan J.; Southam, Andrew D.; MacKay, Hannah L.; Ellington, Max A; Snow, Martyn D.; Farhat, Khanim L.; Bunce, Christopher M.; Johnson, William E. B.; Aston University, Birmingham; University of Birmingham; University Centre Shrewsbury; Royal Orthopaedic Hospital, Birmingham; University of Chester
      Drug repurposing is a cost effective means of targeting new therapies for cancer. We have examined the effects of the repurposed drugs, bezafibrate, medroxyprogesterone acetate and valproic acid on human osteosarcoma cells, i.e., SAOS2 and MG63 compared with their normal cell counterparts, i.e. mesenchymal stem/stromal cells (MSCs). Cell growth, viability and migration were measured by biochemical assay and live cell imaging, whilst levels of lipid-synthesising enzymes were measured by immunoblotting cell extracts. These drug treatments inhibited the growth and survival of SAOS2 and MG63 cells most effectively when used in combination (termed V-BAP). In contrast, V-BAP treated MSCs remained viable with only moderately reduced cell proliferation. V-BAP treatment also inhibited migratory cell phenotypes. MG63 and SAOS2 cells expressed much greater levels of fatty acid synthase and stearoyl CoA desaturase 1 than MSCs, but these elevated enzyme levels significantly decreased in the V-BAP treated osteosarcoma cells prior to cell death. Hence, we have identified a repurposed drug combination that selectively inhibits the growth and survival of human osteosarcoma cells in association with altered lipid metabolism without adversely affecting their non-transformed cell counterparts.
    • Eukarion-134 Attenuates Endoplasmic Reticulum Stress-Induced Mitochondrial Dysfunction in Human Skeletal Muscle Cells

      Nye, Gareth; Thoma, Anastasia; Lyon, Max; Al-Shanti, Nasser; Cooper, Robert; Lightfoot, Adam; University of Chester; Manchester Metropolitan University; University of Liverpool
      Maladaptive endoplasmic reticulum (ER) stress is associated with modified reactive oxygen species (ROS) generation and mitochondrial abnormalities; and is postulated as a potential mechanism involved in muscle weakness in myositis, an acquired autoimmune neuromuscular disease. This study investigates the impact of ROS generation in an in vitro model of ER stress in skeletal muscle, using the ER stress inducer tunicamycin (24 h) in the presence or absence of a superoxide dismutase/catalase mimetic Eukarion (EUK)-134. Tunicamycin induced maladaptive ER stress, which was mitigated by EUK-134 at the transcriptional level. ER stress promoted mitochondrial dysfunction, described by substantial loss of mitochondrial membrane potential, as well as a reduction in respiratory control ratio, reserve capacity, phosphorylating respiration, and coupling efficiency, which was ameliorated by EUK-134. Tunicamycin induced ROS-mediated biogenesis and fusion of mitochondria, which, however, had high propensity of fragmentation, accompanied by upregulated mRNA levels of fission-related markers. Increased cellular ROS generation was observed under ER stress that was prevented by EUK-134, even though no changes in mitochondrial superoxide were noticeable. These findings suggest that targeting ROS generation using EUK-134 can amend aspects of ER stress-induced changes in mitochondrial dynamics and function, and therefore, in instances of chronic ER stress, such as in myositis, quenching ROS generation may be a promising therapy for muscle weakness and dysfunction.
    • Recombinant Plasmodium vivax circumsporozoite surface protein allelic variants: antibody recognition by individuals from three communities in the Brazilian Amazon

      Ferreira Soares, Isabela; López-Camacho, César; Nunes Rodrigues-da-Silva, Rodrigo; da Silva Matos, Ada; de Oliveira Baptista, Barbara; Renato Rivas Totino, Paulo; Medeiros de Souza, Rodrigo; Harrison, Kate; Gimenez, Alba Marina; Oliveira de Freitas, Elisângela; et al.
      Circumsporozoite protein (CSP) variants of P. vivax, besides having variations in the protein repetitive portion, can differ from each other in aspects such as geographical distribution, intensity of transmission, vectorial competence and immune response. Such aspects must be considered to P. vivax vaccine development. Therefore, we evaluated the immunogenicity of novel recombinant proteins corresponding to each of the three P. vivax allelic variants (VK210, VK247 and P. vivax-like) and of the C-terminal region (shared by all PvCSP variants) in naturally malaria-exposed populations of Brazilian Amazon. Our results demonstrated that PvCSP-VK210 was the major target of humoral immune response in studied population, presenting higher frequency and magnitude of IgG response. The IgG subclass profile showed a prevalence of cytophilic antibodies (IgG1 and IgG3), that seem to have an essential role in protective immune response. Differently of PvCSP allelic variants, antibodies elicited against C-terminal region of protein did not correlate with epidemiological parameters, bringing additional evidence that humoral response against this protein region is not essential to protective immunity. Taken together, these findings increase the knowledge on serological response to distinct PvCSP allelic variants and may contribute to the development of a global and effective P. vivax vaccine.
    • Occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants and e-waste soil-derived extracts.

      Eze, Chukwuebuka ThankGod; orcid: 0000-0001-8076-2926; email: thankgod.eze@fuoye.edu.ng; Michelangeli, Francesco; Otitoloju, Adebayo Akeem; Eze, Obianuju Oluchukwu; Ibraheem, Omodele; Ogbuene, Emeka Bright; Ogunwole, Germaine Akinola (2020-10-25)
      We investigated the occurrence of chemical pollutants in major e-waste sites in West Africa and usefulness of cytotoxicity and induction of ethoxyresorufin-O-deethylase (EROD) in determining the effects of some detected brominated flame retardants (BFRs) and e-waste soil-derived extracts. Analysis of the e-waste site samples using AAS and GC-MS techniques revealed the presence of a range of toxic metals as well as persistent and toxic organic pollutants, respectively, in the vicinity of the e-waste sites. As expected, the occurrence (%) of all the detected chemical pollutants in experimental soils significantly (P < 0.05) differs from occurrence (%) in control soil. The calculated LC values on RBL-2H3 cells of the detected tetrabromobisphenol A (TBBPA) and hexabromocyclododecane (HBCD) were 3.75 μM and 4.2 μM, respectively. Tribromophenol (TBP), dibromobiphenyl (DBB), and decabromodiphenyl ether (DBDE) were remarkably less toxic on RBL-2H3 cells compared with TBBPA and HBCD as they did not reduce RBL-2H3 cell viability below 50% in the tested concentration range (0-20 μM). The study revealed that TBBPA and HBCD could induce significant RBL-2H3 cell death through caspase-dependent apoptosis. The study further shows that the cytotoxicity of some of these BFRs could increase synergistically when in mixtures and potentially activate inflammation through the stimulation of mast cell degranulation. The e-waste soil-derived extracts induced a concentration-dependent increase in EROD activity in the exposed RTG-W1 cells. Ultimately, nonpolar extracts had higher EROD-inducing potency compared with polar extracts and hence suggesting the presence in higher amounts of AhR agonists in nonpolar e-waste soil-derived extracts than polar extracts. Overall, there is urgent need for actions in order to improve the environmental quality of the e-waste sites.
    • The efficacy of using Appropriate Paper-based Technology postural support devices in Kenyan children with Cerebral Palsy

      Barton, Catherine; Buckley, John P.; Samia, Pauline; Williams, Fiona; Taylor, Sue; Lindoewood, Rachel; University Centre Shrewsbury - University of Chester
      Purpose: Appropriate paper-based technology (APT) is used to provide postural support for children with cerebral palsy (CP) in low-resourced settings. This pilot study aimed to evaluate the impact of APT on the children’s and families’ lives. Materials and methods: A convenience sample of children with CP and their families participated. Inclusion was based on the Gross Motor Function Classification System levels IV and V. APT seating or standing frames were provided for six months. A mixed methods impact of APT devices on the children and families included the Family Impact Assistive Technology Scale for Adaptive Seating (FIATS-AS); the Child Engagement in Daily Life (CEDL) questionnaire; and a qualitative assessment from diary/log and semi-structured interviews. Results: Ten children (median 3 years, range 9 months - 7 years). Baseline to follow-up median (IQR) FIATS-AS were: 22.7 (9.3) and 30.3 (10.2), respectively (p = 0.002). Similarly mean (SD) CEDL scores for “frequency” changed from 30.5 (13.2) to 42.08 (5.96) (p=0.021) and children’s enjoyment scores from 2.23 (0.93) to 2.91 (0.79) (p = 0.019). CEDL questionnaire for self-care was not discriminatory; seven families scored zero at both baseline and 6 months. Qualitative interviews revealed three key findings; that APT improved functional ability, involvement/interaction in daily-life situations, and a reduced family burden of care. Conclusion: APT devices used in Kenyan children with non-ambulant CP had a meaningful positive effect on both the children’s and their families’ lives.
    • History of traction

      Flynn, Sandra (Elsevier, 2020-10-01)
    • An in vitro comparison of the neurotrophic and angiogenic activity of human and canine adipose-derived mesenchymal stem cells (MSCs): translating MSC-based therapies for spinal cord injury.

      Al Delfi, Ibtesam R. T; Wood, Chelsea R.; Johnson, Louis D. V.; Snow, Martyn D.; Innes, John F.; Myint, Peter; Johnson, William E. B.; Kut University College; University of Chester; Royal Orthopaedic Hospital, Birmingham; Veterinary Tissue Bank, Chirk
      The majority of research into the effects of mesenchymal stem cell (MSC) transplants on spinal cord injury (SCI) is performed in rodent models, which may help inform on mechanisms of action, but does not represent the scale and wound heterogeneity seen in human SCI. In contrast, SCI in dogs occurs naturally, is more akin to human SCI, and can be used to help address important aspects of the development of human MSC-based therapies. To enable translation to the clinic and a comparison across species, we have examined the paracrine, regenerative capacity of human and canine adipose-derived MSCs in vitro. MSCs were initially phenotyped according to tissue culture plastic adherence, CD immunoprofiling and tri-lineage differentiation potential. Conditioned medium (CM) from MSC cultures was then assessed for its neurotrophic and angiogenic activity using established cell-based assays. MSC CM significantly increased neuronal cell proliferation, neurite outgrowth, and βIII tubulin immunopositivity. In addition, MSC CM significantly increased endothelial cell migration, cell proliferation and the formation of tubule-like structures in Matrigel assays. There were no marked or significant differences in the capacity of human or canine MSC CM to stimulate neuronal cell or endothelial cell activity. Hence, this study supports the use of MSC transplants for canine SCI, furthermore it increases understanding of how this may subsequently provide useful information and translate to MSC transplants for human SCI.
    • Nicotinamide restricts neural precursor proliferation to enhance catecholaminergic neuronal subtype differentiation from mouse embryonic stem cells

      Borlongan, Cesar V.; Griffin, Síle M.; orcid: 0000-0002-6670-5084; email: silemgriffin@gmail.com; Pickard, Mark R.; Hawkins, Clive P.; Williams, Adrian C.; Fricker, Rosemary A.; orcid: 0000-0001-8768-510X (Public Library of Science, 2020-09-14)
      Emerging evidence indicates that a strong relationship exists between brain regenerative therapies and nutrition. Early life nutrition plays an important role during embryonic brain development, and there are clear consequences to an imbalance in nutritional factors on both the production and survival of mature neuronal populations and the infant’s risk of diseases in later life. Our research and that of others suggest that vitamins play a fundamental role in the formation of neurons and their survival. There is a growing body of evidence that nicotinamide, the water-soluble amide form of vitamin B3, is implicated in the conversion of pluripotent stem cells to clinically relevant cells for regenerative therapies. This study investigated the ability of nicotinamide to promote the development of mature catecholaminergic neuronal populations (associated with Parkinson’s disease) from mouse embryonic stem cells, as well as investigating the underlying mechanisms of nicotinamide’s action. Nicotinamide selectively enhanced the production of tyrosine hydroxylase-expressing neurons and serotonergic neurons from mouse embryonic stem cell cultures (Sox1GFP knock-in 46C cell line). A 5-Ethynyl-2´-deoxyuridine (EdU) assay ascertained that nicotinamide, when added in the initial phase, reduced cell proliferation. Nicotinamide drove tyrosine hydroxylase-expressing neuron differentiation as effectively as an established cocktail of signalling factors, reducing the proliferation of neural progenitors and accelerating neuronal maturation, neurite outgrowth and neurotransmitter expression. These novel findings show that nicotinamide enhanced and enriched catecholaminergic differentiation and inhibited cell proliferation by directing cell cycle arrest in mouse embryonic stem cell cultures, thus driving a critical neural proliferation-to-differentiation switch from neural progenitors to neurons. Further research into the role of vitamin metabolites in embryogenesis will significantly advance cell-based regenerative medicine, and help realize their role as crucial developmental signalling molecules in brain development.
    • Comparing physician associates and foundation year two doctors-in-training undertaking emergency medicine consultations in England: a mixed-methods study of processes and outcomes

      Halter, Mary; orcid: 0000-0001-6636-0621; Drennan, Vari; orcid: 0000-0002-8915-5185; Wang, Chao; Wheeler, Carly; Gage, Heather; Nice, Laura; de Lusignan, Simon; orcid: 0000-0001-5613-6810; Gabe, Jonathan; Brearley, Sally; Ennis, James; et al. (BMJ Publishing Group, 2020-09-01)
      Objectives: To compare the contribution of physician associates to the processes and outcomes of emergency medicine consultations with that of foundation year two doctors-in-training. Design: Mixed-methods study: retrospective chart review using 4 months’ anonymised clinical record data of all patients seen by physician associates or foundation year two doctors-in-training in 2016; review of a subsample of 40 records for clinical adequacy; semi-structured interviews with staff and patients; observations of physician associates. Setting: Three emergency departments in England. Participants: The records of 8816 patients attended by 6 physician associates and 40 foundation year two doctors-in-training; of these n=3197 had the primary outcome recorded (n=1129 physician associates, n=2068 doctor); 14 clinicians and managers and 6 patients or relatives for interview; 5 physician associates for observation. Primary and secondary outcome measures: The primary outcome was unplanned re-attendance at the same emergency department within 7 days. Secondary outcomes: consultation processes, clinical adequacy of care, and staff and patient experience. Results: Re-attendances within 7 days (n=194 (6.1%)) showed no difference between physician associates and foundation year two doctors-in-training (OR 0.87, 95% CI 0.61 to 1.24, p=0.437). If seen by a physician associate, patients were more likely receive an X-ray investigation (OR 2.10, 95% CI 1.72 to 4.24), p<0.001), after adjustment for patient characteristics, triage severity of condition and statistically significant clinician intraclass correlation. Clinical reviewers found almost all patients’ charts clinically adequate. Physician associates were evaluated as assessing patients in a similar way to foundation year two doctors-in-training and providing continuity in the team. Patients were positive about the care they had received from a physician associate, but had poor understanding of the role. Conclusions: Physician associates in emergency departments in England treated patients with a range of conditions safely, and at a similar level to foundation year two doctors-in-training, providing clinical operational efficiencies.
    • 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
    • Early Transplantation of Mesenchymal Stem Cells After Spinal Cord Injury Relieves Pain Hypersensitivity Through Suppression of Pain-Related Signaling Cascades and Reduced Inflammatory Cell Recruitment

      Johnson, William Eustace Basil; Watanabe, Shuji; Uchida, Kenzo; Nakajima, Hideaki; Matsuo, Hideaki; Sugita, Daisuke; Yoshida, Ai; Honjoh, Kazuya; Baba, Hisatoshi; Aston University, University of Fukui
      Bone marrow-derived mesenchymal stem cells (BMSC) modulate inflammatory/immune responses and promote motor functional recovery after spinal cord injury (SCI). However, the effects of BMSC transplantation on central neuropathic pain and neuronal hyperexcitability after SCI remain elusive. This is of importance because BMSC-based therapies have been proposed for clinical treatment. We investigated the effects of BMSC transplantation on pain hypersensitivity in green fluorescent protein (GFP)-positive bone marrow-chimeric mice subjected to a contusion SCI, and the mechanisms of such effects. BMSC transplantation at day 3 post-SCI improved motor function and relieved SCI-induced hypersensitivities to mechanical and thermal stimulation. The pain improvements were mediated by suppression of protein kinase C-γ and phosphocyclic AMP response element binding protein expression in dorsal horn neurons. BMSC transplants significantly reduced levels of p-p38 mitogen-activated protein kinase and extracellular signal-regulated kinase (p-ERK1/2) in both hematogenous macrophages and resident microglia and significantly reduced the infiltration of CD11b and GFP double-positive hematogenous macrophages without decreasing the CD11b-positive and GFP-negative activated spinal-microglia population. BMSC transplants prevented hematogenous macrophages recruitment by restoration of the blood-spinal cord barrier (BSCB), which was associated with decreased levels of (a) inflammatory cytokines (tumor necrosis factor-α, interleukin-6); (b) mediators of early secondary vascular pathogenesis (matrix metallopeptidase 9); (c) macrophage recruiting factors (CCL2, CCL5, and CXCL10), but increased levels of a microglial stimulating factor (granulocyte-macrophage colony-stimulating factor). These findings support the use of BMSC transplants for SCI treatment. Furthermore, they suggest that BMSC reduce neuropathic pain through a variety of related mechanisms that include neuronal sparing and restoration of the disturbed BSCB, mediated through modulation of the activity of spinal-resident microglia and the activity and recruitment of hematogenous macrophages.
    • The developing landscape of diagnostic and prognostic biomarkers for spinal cord injury in cerebrospinal fluid and blood

      Hulme CH; Brown SJ; Fuller HR; Riddell J; Osman A; Chowdhury J; Kumar N; Johnson WE; Wright KT; Keele University, RJAH Orthopaedic Hospital, University of Glasgow, University of Chester (Nature Publishing Group, 2016-12-20)
      STUDY DESIGN: Review study. OBJECTIVES: The identification of prognostic biomarkers of spinal cord injury (SCI) will help to assign SCI patients to the correct treatment and rehabilitation regimes. Further, the detection of biomarkers that predict permanent neurological outcome would aid in appropriate recruitment of patients into clinical trials. The objective of this review is to evaluate the current state-of-play in this developing field. SETTING: Studies from multiple countries were included. METHODS: We have completed a comprehensive review of studies that have investigated prognostic biomarkers in either the blood or cerebrospinal fluid (CSF) of animals and humans following SCI. RESULTS: Targeted and unbiased approaches have identified several prognostic biomarkers in CSF and blood. These proteins associate with cellular damage following SCI and include components from neurons, oligodendrocytes and reactive astrocytes, that is, neurofilament proteins, glial fibrillary acidic protein, Tau and S100 calcium-binding protein β. Unbiased approaches have also identified microRNAs that are specific to SCI, as well as other cell damage-associated proteins. CONCLUSIONS: The discovery and validation of stable, specific, sensitive and reproducible biomarkers of SCI is a rapidly expanding field of research. So far, few studies have utilised unbiased approaches aimed at the discovery of biomarkers within the CSF or blood in this field; however, some targeted approaches have been successfully used. Several studies using various animal models and some with small human patient cohorts have begun to pinpoint biomarkers in the CSF and blood with putative prognostic value. An increased sample size will be required to validate these biomarkers in the heterogeneous clinical setting.
    • The influence of pH and fluid dynamics on the antibacterial efficacy of 45S5 Bioglass Short title: Antibacterial efficacy of 45S5 Bioglass

      Begum, Saima; Johnson, William Eustace Basil; Worthington, Tony; Martin, Richard; Aston University (IOP Publishing, 2016-02-02)
      In recent years, there has been considerable interest in the potential antibacterial properties that bioactive glasses may possess. However, there have been several conflicting reports on the antibacterial efficacy of 45S5 Bioglass®. Various mechanisms regarding its mode of action have been proposed, such as changes in the environmental pH, increased osmotic pressure, and 'needle-like' sharp glass debris which could potentially damage prokaryotic cell walls and thus inactivate bacteria. In this current study, a systematic investigation was undertaken on the antibacterial efficacy of 45S5 Bioglass® on Escherichia coli NCTC 10538 and Staphylococcus aureus ATCO 6538 under a range of clinically relevant scenarios including varying Bioglass® concentration, direct and indirect contact between Bioglass® and microorganisms, static and shaking incubation conditions, elevated and neutralised pH environments. The results demonstrated that, under elevated pH conditions, Bioglass® particles have no antibacterial effect on S. aureus while a concentration dependent antibacterial effect against E. coli was observed. However, the antibacterial activity ceased when the pH of the media was neutralised. The results of this current study, therefore, suggest that the mechanism of antibacterial activity of Bioglass® is associated with changes in the environmental pH; an environment that is less likely to occur in vivo due to buffering of the system.
    • Bone Marrow-Derived Mesenchymal Stem Cells Become Antiangiogenic When Chondrogenically or Osteogenically Differentiated: Implications for Bone and Cartilage Tissue Engineering

      Bara, Jennifer; Johnson, William Eustace Basil; Roberts, Sally; McCarthy, Helen E.; Humphrey, Emma; AO Institute, Davos, Switzerland; Aston University; Keele University
      Osteochondral tissue repair requires formation of vascularized bone and avascular cartilage. Mesenchymal stem cells stimulate angiogenesis both in vitro and in vivo but it is not known if these proangiogenic properties change as a result of chondrogenic or osteogenic differentiation. We investigated the angiogenic/antiangiogenic properties of equine bone marrow-derived mesenchymal stem cells (eBMSCs) before and after differentiation in vitro. Conditioned media from chondrogenic and osteogenic cell pellets and undifferentiated cells was applied to endothelial tube formation assays using Matrigel. Additionally, the cell secretome was analysed using LC-MS/MS mass spectrometry and screened for angiogenesis and neurogenesis-related factors using protein arrays. Endothelial tube-like formation was supported by conditioned media from undifferentiated eBMSCs. Conversely, chondrogenic and osteogenic conditioned media was antiangiogenic as shown by significantly decreased length of endothelial tube-like structures and degree of branching compared to controls. Undifferentiated cells produced higher levels of angiogenesis-related proteins compared to chondrogenic and osteogenic pellets. In summary, eBMSCs produce an array of angiogenesis-related proteins and support angiogenesis in vitro via a paracrine mechanism. However, when these cells are differentiated chondrogenically or osteogenically, they produce a soluble factor(s) that inhibits angiogenesis. With respect to osteochondral tissue engineering, this may be beneficial for avascular articular cartilage formation but unfavourable for bone formation where a vascularized tissue is desired.
    • CD271-selected mesenchymal stem cells from adipose tissue enhance cartilage repair and are less angiogenic than plastic adherent mesenchymal stem cells

      Kohli, Nupur; Johnson, William Eustace Basil; Uchida, Kenzo; Aston University, University of Chester, University of Fukui (Nature, 2019-02-28)
      CD271 is a marker of bone marrow MSCs with enhanced differentiation capacity for bone or cartilage repair. However, the nature of CD271+ MSCs from adipose tissue (AT) is less well understood. Here, we investigated the differentiation, wound healing and angiogenic capacity of plastic adherent MSCs (PA MSCs) versus CD271+ MSCs from AT. There was no difference in the extent to which PA MSCs and CD271+ MSCs formed osteoblasts, adipocytes or chondrocytes in vitro. In contrast, CD271+ MSCs transplanted into athymic rats significantly enhanced osteochondral wound healing with reduced vascularisation in the repair tissue compared to PA MSCs and control animals; there was little histological evidence of mature articular cartilage formation in all animals. Conditioned medium from CD271+ MSC cultures was less angiogenic than PA MSC conditioned medium, and had little effect on endothelial cell migration or endothelial tubule formation in vitro. The low angiogenic activity of CD271+ MSCs and improved early stage tissue repair of osteochondral lesions when transplanted, along with a comparable differentiation capacity along mesenchymal lineages when induced, suggests that these selected cells are a better candidate than PA MSCs for the repair of cartilaginous tissue.