• Lamin A/C dysregulation contributes to cardiac pathology in a mouse model of severe spinal muscular atrophy

      Soltic, Darija; Shorrock, Hannah K; Allardyce, Hazel; Wilson, Emma L; Holt, Ian; Synowsky, Silvia A; Shirran, Sally L; Parson, Simon H; Gillingwater, Thomas H; Fuller, HR; et al.
      Cardiac pathology is emerging as a prominent systemic feature of spinal muscular atrophy (SMA), but little is known about the underlying molecular pathways. Using quantitative proteomics analysis, we demonstrate widespread molecular defects in heart tissue from the Taiwanese mouse model of severe SMA. We identify increased levels of lamin A/C as a robust molecular phenotype in the heart of SMA mice and show that lamin A/C dysregulation is also apparent in SMA patient fibroblast cells and other tissues from SMA mice. Lamin A/C expression was regulated in vitro by knockdown of the E1 ubiquitination factor ubiquitin-like modifier activating enzyme 1, a key downstream mediator of SMN-dependent disease pathways, converging on β-catenin signaling. Increased levels of lamin A are known to increase the rigidity of nuclei, inevitably disrupting contractile activity in cardiomyocytes. The increased lamin A/C levels in the hearts of SMA mice therefore provide a likely mechanism explaining morphological and functional cardiac defects, leading to blood pooling. Therapeutic strategies directed at lamin A/C may therefore offer a new approach to target cardiac pathology in SMA.
    • The long non-coding RNA NEAT1 regulates cell survival in breast cancer cell lines

      Almnaseer, Zainab; Pickard, Mark R.; Mourtada-Maarabouni, Mirna; Keele University, United Kingdom (NCRI Cancer Conference 2015 Abstracts, 2015)
      Background Nuclear long non-coding RNAs (LncRNAs) regulate various cellular processes including the organization of nuclear sub-structures, the alteration of chromatin state, and the regulation of gene expression. Nuclear Enriched Abundant Transcript 1 (NEAT1) is a nuclear lncRNA transcribed from chromosome 11q13. Two transcripts are produced from the NEAT1 gene, 3.7-kb NEAT1_v1 and 23-kb NEAT1_v2. Both isoforms participate in the formation of the nuclear paraspeckles . NEAT1 is reported to be overexpressed in prostate cancer and a direct transcriptional target of hypoxia-inducible factor in many breast cancer cell lines. The aims of this study were to determine the effects of silencing NEAT1 on breast cancer cell survival. Method MCF7 and MDA-MB 231 cells were transfected with siRNAs to different NEAT1 sequences or NEAT1 antisense oligonucleotides (ASO). Controls received scrambled siRNA or scrambled oligonucleotide, as appropriate. In some experiments, cells were exposed to ultraviolet-C (UV-C) light post-transfection to induce apoptosis, and then culture viability and apoptosis were assessed. NEAT1 expression was evaluated by qRT-PCR TaqMan® analysis. Results In MCF7 and MDA-MB-231 cells, siRNA-mediated silencing of NEAT1 reduced basal survival and after UV-C irradiation and decreased their colony forming ability. NEAT1 ASOs were more effective in silencing NEAT1 and caused a greater reduction in cell viability. NEAT1 silencing also affected cell cycle profile by enhancing the proportion of cells in G0/G1 phase. Conclusion NEAT1 regulates the survival of Breast cells. Down regulation of NEAT1 expression decreased cell survival, proliferation and modulated cell cycle progression of breast cancer cells, indicating a link between the NEAT1 expression levels and carcinogenesis of breast cancer.
    • Long non-coding RNAs: new opportunities and old challenges in cancer therapy

      Williams, Gwyn T.; Pickard, Mark R.; Keele University; University of Chester (AME Publishing Company, 2016-09)
      No abstract - invited commentary
    • Long-term administration of the mitochondria-targeted antioxidant mitoquinone mesylate fails to attenuate age-related oxidative damage or rescue the loss of muscle mass and function associated with aging of skeletal muscle

      Nye, Gareth; Sakellariou, Giorgos; Lightfoot, Adam; Pearson, Timothy; Wells, Nicola; McArdle, Anne; Jackson, Malcolm; Giakoumaki, Ifigeneia; Griffiths, Richard; University of Liverpool (Faseb Journal, 2016-08-22)
      Age-related skeletal muscle dysfunction is the underlying cause of morbidity that affects up to half the population aged 80 and over. Considerable evidence indicates that oxidative damage and mitochondrial dysfunction contribute to the sarcopenic phenotype that occurs with aging. To examine this, we administered the mitochondria-targeted antioxidant mitoquinone mesylate {[10-(4,5-dimethoxy-2-methyl-3,6-dioxo-1,4-cyclohexadien-1-yl)decyl] triphenylphosphonium; 100 μM} to wild-type C57BL/6 mice for 15 wk (from 24 to 28 mo of age) and investigated the effects on age-related loss of muscle mass and function, changes in redox homeostasis, and mitochondrial organelle integrity and function. We found that mitoquinone mesylate treatment failed to prevent age-dependent loss of skeletal muscle mass associated with myofiber atrophy or alter a variety of in situ and ex vivo muscle function analyses, including maximum isometric tetanic force, decline in force after a tetanic fatiguing protocol, and single-fiber-specific force. We also found evidence that long-term mitoquinone mesylate administration did not reduce mitochondrial reactive oxygen species or induce significant changes in muscle redox homeostasis, as assessed by changes in 4-hydroxynonenal protein adducts, protein carbonyl content, protein nitration, and DNA damage determined by the content of 8-hydroxydeoxyguanosine. Mitochondrial membrane potential, abundance, and respiration assessed in permeabilized myofibers were not significantly altered in response to mitoquinone mesylate treatment. Collectively, these findings demonstrate that long-term mitochondria-targeted mitoquinone mesylate administration failed to attenuate age-related oxidative damage in skeletal muscle of old mice or provide any protective effect in the context of muscle aging
    • Low leukotriene B4 receptor 1 leads to ALOX5 downregulation at diagnosis of chronic myeloid leukemia

      Lucas, Claire; Harris, Robert; McDonald, Elizabeth; Giannoudis, Athina; Clark, Richard; University of Liverpool, Royal Liverpool University hospital, (Ferrata Storti Foundation, 2014-11-01)
      ALOX5 is implicated in chronic myeloid leukemia development in mouse leukemic stem cells, but its importance in human chronic myeloid leukemia is unknown. Functional ALOX5 was assessed using an LTB4 ELISA and ALOX5, and LTB4R1 mRNA expression was determined via a TaqMan gene expression assay. LTB4R1 and 5-LOX protein levels were assessed by cell surface flow cytometry analysis. At diagnosis ALOX5 was below normal in both blood and CD34(+) stem cells in all patients. On treatment initiation, ALOX5 levels increased in all patients except those who were destined to progress subsequently to blast crisis. LTB4 levels were increased despite low ALOX5 expression, suggesting that the arachidonic acid pathway is functioning normally up to the point of LTB4 production. However, the LTB4 receptor (BLT1) protein in newly diagnosed patients was significantly lower than after a period of treatment (P<0.0001). The low level of LTB4R1 at diagnosis explains the downregulation of ALOX5. In the absence of LTB4R1, the arachidonic acid pathway intermediates (5-HEPTE and LTA4) negatively regulate ALOX5. ALOX5 regulation is aberrant in chronic myeloid leukemia patients and may not be important for the development of the disease. Our data suggest caution when extrapolating mouse model data into human chronic myeloid leukemia.
    • Magnetic nanoparticle-mediated gene delivery to two- and three-dimensional neural stem cell cultures: magnet-assisted transfection and multifection approaches to enhance outcomes

      Pickard, Mark R.; Adams, Christopher F.; Chari, Divya M.; University of Chester; Keele University (Wiley, 2017-02-02)
      Neural stem cells (NSCs) have high translational potential in transplantation therapies for neural repair. Enhancement of their therapeutic capacity by genetic engineering is an important goal for regenerative neurology. Magnetic nanoparticles (MNPs) are major non-viral vectors for safe bioengineering of NSCs, offering critical translational benefits over viral vectors, including safety, scalability, and ease of use. This unit describes protocols for the production of suspension (neurosphere) and adherent (monolayer) murine NSC cultures. Genetic engineering of NSCs with MNPs and the application of 'magnetofection' (magnetic fields) or 'multifection' (repeat transfection) approaches to enhance gene delivery are described. Magnetofection of monolayer cultures achieves optimal transfection, but neurospheres offer key advantages for neural graft survival post-transplantation. A protocol is presented which allows the advantageous features of each approach to be combined into a single procedure for transplantation. The adaptation of these protocols for other MNP preparations is considered, with emphasis on the evaluation of procedural safety.
    • Manipulation of apoptosis in cancer cells

      Williams, John H. H.; Ireland, Elyse; Sahib, Muneera M (University of Chester, 2018-09-18)
      Conventional cancer therapies can have severe side effects, so new strategies to limit these needs to be investigated. Several anticancer agents induce the expression of tumour suppressor gene p21 in colorectal cancer cell line HT-29. Interestingly, the stress protein HSPA1A is also often elevated in tumour cells and has an anti - apoptotic activity. The main aim of this study was to examine whether a two - pronged approach, overexpressing p21 (using genetic approach and inhibition of HSPA1A using pifithrin - µ would be effective in inducing apoptosis in tumour cells. Chitosan or BSA based delivery systems were evaluated for cytotoxicity, with the intension of using it for plasmid DNA based cell transfections in this study. The interaction of HSPA1A protein in combination treatments involving UV radiation and hyperthermia at 42℃ were also evaluated to perceive the various roles of HSPA1A in arresting colorectal cancer cells. Colorectal cancer cell lines HT-29 and leukaemia cancer cell lines U937 were used in the study. All experiments were performed with cancer cell lines maintained in culture medium devoid of antibiotics. Cell cytotoxicity were evaluated using MTS and PI assays. The rate of apoptosis was determined using annexin V and PI staining by flow cytometry. Chitosan or BSA based microparticles or microgels were observed for size determination or morphology using scanning electron microscopy. Full length human p21 inserted plasmid DNA was a gift from Mien - Chie Hung, Addgene, USA. HT- 29 cells were subjected to p21 plasmid DNA transfection effects. Cells were treated with pifithrin - µ (15µM) prior to gene transfection to address its combined effect with p21 plasmid DNA transfection. HSPA1A and p21 protein expression studies were analysed using FITC labelled antibodies by flow cytometer. Combination studies with HSPA1A inhibitor pifithrin - µ and UV reflected enhanced cytotoxicity compared with either of the treatments independently. Hyperthermia at 42℃ induced apoptosis by MTS assay, which was confirmed by flow cytometric analysis in both the cell lines tested. Considering the cytotoxicity reflected by the chitosan or BSA delivery systems in drug free states, the p21 plasmid DNA transfection was carried out using lipofectamine 2000. Both overexpression of p21 and inhibition of HSPA1A protein with pifithrin - µ enhanced the rate of apoptosis with statistical significance of (p-<0.0001****) compared to the respected controls. The data in this thesis suggests the inhibition of HSPA1A in combination with increased p21 would be a promising therapeutic strategy for the treatment of colorectal cancers.
    • Mechanisms of skeletal muscle ageing: avenues for therapeutic intervention

      Nye, Gareth; McCormick, Rachel; Lightfoot, Adam; McArdle, Anne; University of Liverpool (Elsevier, 2014-05-28)
      Age-related loss of muscle mass and function, termed sarcopenia, is a catastrophic process, which impacts severely on quality of life of older people. The mechanisms underlying sarcopenia are unclear and the development of optimal therapeutic interventions remains elusive. Impaired regenerative capacity, attenuated ability to respond to stress, elevated reactive oxygen species production and low-grade systemic inflammation are all key contributors to sarcopenia. Pharmacological intervention using compounds such as 17AAG, SS-31 and Bimagrumab or naturally occurring polyphenols to target specific pathways show potential benefit to combat sarcopenia although further research is required, particularly to identify the mechanisms by which muscle fibres are completely lost with increasing age.
    • Mental Health Decisions; what every officer should consider

      Williams, Barry; Jones, Steven; University of Chester (Police Professional, 2012-05-24)
      It can often appear to Police officers that they are damned if they do make decisions, and damned if they don’t in mental health cases. A culture has evolved that triggers decision apathy and defensive decisions that arguably do not benefit the Police, public, or the mental health arrestee. Decisions of this presenting complexity in whatever profession must be made and firmly rooted within the current evidence base, lawful, and also be reasonable in the given situation. It is therefore not unreasonable to expect officers to explain and account for how and why they acted as they did, and the frameworks (statutes/ codes) which should underpin such practice decisions. It is of paramount importance that Police officers are kept appraised of developments in mental health cases and how this crucially will inform, and sometimes correct custom and practice. This article in three parts aims firstly to refresh officer’s knowledge. Second, inform current practice and address practice from recent cases involving the police and mental health patients. Thirdly, and perhaps the most crucial through case examples offer a decision making framework to support operational staff in the right direction for mental health practice and defend practice challenges that may arise at all levels.
    • Methadone‐Assisted Opiate Withdrawal and Subsequent Heroin Abstinence: The Importance of Psychological Preparedness

      Jones, Steven; Jack, Barbara; Kirby, Julie; Wilson, Thomas; Murphy, Philip; University of Chester
      Background and Objectives: Treatment guidelines emphasize patients’ readiness for transitioning from opiate substitution treatment (OST) to opiate withdrawal and abstinence. Psychological preparedness indicators for this transition were examined. Methods: Patients (all male) were recruited from three treatment settings: prison, an inpatient detoxification unit, and an outpatient clinic. Time 1 (T1) was admission to methadone‐assisted withdrawal in all settings. Time 2 (T2) was a 6‐month follow‐up. With n = 24 at T1 for each group (N = 72), a battery of instruments relevant to psychological preparedness was administered. Results: At T1, inpatients had higher self‐efficacy beliefs for successful treatment completion than prison patients. For patients contactable at T2, T1 self‐efficacy positively predicted T2 opiate abstinence. No other variable improved prediction. T1 SOCRATES (Stages of Change Readiness and Treatment Eagerness Scale) ambivalence scores, age, and lifetime heroin use duration predicted maintenance of contact or not with treatment services and contactability by the researcher. Measures of mood did not differ between groups at T1 or predict T2 outcomes. Discussion and Conclusions: Self‐efficacy beliefs are a potentially useful indicator of readiness for transitioning from OST to a medically assisted opiate withdrawal and subsequent abstinence. Ambivalence regarding change, age, and lifetime heroin use duration are potentially useful predictors of patients maintaining contact with services, and of being retained in research. Scientific Significance: These findings advance existing literature and knowledge by highlighting the importance of self‐efficacy in psychological preparedness for opiate abstinence, and the predictive utility to clinicians of this and other variables measurable at admission, in the clinical management of opiate users
    • Mitochondrial ROS regulate oxidative damage and mitophagy but not age-related muscle fiber atrophy

      Nye, Gareth; Sakellariou, Giorgos; Pearson, Timothy; Lightfoot, Adam; Wells, Nicola; Giakoumaki, Ifigeneia; Vasilaki, Aphrodite; Griffiths, Richard; Jackson, Malcolm; McArdle, Anne; et al. (Nature Research, 2016-09-29)
      Age-related loss of skeletal muscle mass and function is a major contributor to morbidity and has a profound effect on the quality of life of older people. The potential role of age-dependent mitochondrial dysfunction and cumulative oxidative stress as the underlying cause of muscle aging remains a controversial topic. Here we show that the pharmacological attenuation of age-related mitochondrial redox changes in muscle with SS31 is associated with some improvements in oxidative damage and mitophagy in muscles of old mice. However, this treatment failed to rescue the age-related muscle fiber atrophy associated with muscle atrophy and weakness. Collectively, these data imply that the muscle mitochondrial redox environment is not a key regulator of muscle fiber atrophy during sarcopenia but may play a key role in the decline of mitochondrial organelle integrity that occurs with muscle aging.
    • Molecular and Cellular Mechanisms of Action of Tumour Suppressor GAS5 LncRNA

      Pickard, Mark R.; Williams, Gwyn T.; Keele University (MDPI, 2015-07-07)
      It is increasingly recognised that lncRNAs play essential regulatory roles in fundamental biological processes and, consequently, that their dysregulation may contribute to major human diseases, including cancer. Better understanding of lncRNA biology may therefore offer new insights into pathogenetic mechanisms and thereby offer novel opportunities for diagnosis and therapy. Of particular interest in this regard is GAS5 lncRNA, which is down-regulated in multiple cancers, with expression levels related to both clinico-pathological characteristics and patient prognosis. Functional studies have further shown that GAS5 lncRNA both inhibits the proliferation and promotes the apoptosis of multiple cell types, and that together these cellular mechanisms of action are likely to form the basis of its tumour suppressor action. At the same time, advances have been made in our understanding of the molecular mechanisms of GAS5 lncRNA action in recent years, including riborepression of certain steroid hormone receptors and sequestration of miR-21, impacting key regulatory pathways of cell survival. Overall this accumulating knowledge has the potential to improve both the diagnosis and treatment of cancer, and ultimately patient outcome.
    • Muscling in on mitochondrial sexual dimorphism; role of mitochondrial dimorphism in skeletal muscle health and disease

      Nye, Gareth; Lightfoot, Adam; Sakellariou, Giorgos; Degans, Hans; University of Manchester, Manchester Metropolitan University (Portland Press, 2017-07-07)
      Mitochondria are no longer solely regarded as the cellular powerhouse; instead, they are now implicated in mediating a wide-range of cellular processes, in the context of health and disease. A recent article in Clinical Science, Ventura-Clapier et al. highlights the role of sexual dimorphism in mitochondrial function in health and disease. However, we feel the authors have overlooked arguably one of the most mitochondria-rich organs in skeletal muscle. Many studies have demonstrated that mitochondria have a central role in mediating the pathogenesis of myopathologies. However, the impact of sexual dimorphism in this context is less clear, with several studies reporting conflicting observations. For instance in ageing studies, a rodent model reported female muscles have higher antioxidant capacity compared with males; in contrast, human studies demonstrate no sex difference in mitochondrial bioenergetics and oxidative damage. These divergent observations highlight the importance of considering models and methods used to examine mitochondrial function, when interpreting these data. The use of either isolated or intact mitochondrial preparations in many studies appears likely to be a source of discord, when comparing many studies. Overall, it is now clear that more research is needed to determine if sexual dimorphism is a contributing factor in the development of myopathologies.
    • Neferine induces autophagy-dependent cell death in apoptosis-resistant cancers via ryanodine receptor and Ca

      Law, Betty Yuen Kwan; Michelangeli, Francesco; Qu, Yuan Qing; orcid: 0000-0003-3733-3661; Xu, Su-Wei; Han, Yu; Mok, Simon Wing Fai; Dias, Ivo Ricardo De Seabra Rodrigues; Javed, Masood-Ul-Hassan; Chan, Wai-Kit; Xue, Wei-Wei; et al. (2019-12-27)
      Resistance of cancer cells to chemotherapy is a significant clinical concern and mechanisms regulating cell death in cancer therapy, including apoptosis, autophagy or necrosis, have been extensively investigated over the last decade. Accordingly, the identification of medicinal compounds against chemoresistant cancer cells via new mechanism of action is highly desired. Autophagy is important in inducing cell death or survival in cancer therapy. Recently, novel autophagy activators isolated from natural products were shown to induce autophagic cell death in apoptosis-resistant cancer cells in a calcium-dependent manner. Therefore, enhancement of autophagy may serve as additional therapeutic strategy against these resistant cancers. By computational docking analysis, biochemical assays, and advanced live-cell imaging, we identified that neferine, a natural alkaloid from Nelumbo nucifera, induces autophagy by activating the ryanodine receptor and calcium release. With well-known apoptotic agents, such as staurosporine, taxol, doxorubicin, cisplatin and etoposide, utilized as controls, neferine was shown to induce autophagic cell death in a panel of cancer cells, including apoptosis-defective and -resistant cancer cells or isogenic cancer cells, via calcium mobilization through the activation of ryanodine receptor and Ulk-1-PERK and AMPK-mTOR signaling cascades. Taken together, this study provides insights into the cytotoxic mechanism of neferine-induced autophagy through ryanodine receptor activation in resistant cancers.
    • Neuropsychiatric symptoms following metal-on-metal implant failure with cobalt and chromium toxicity

      Green, Ben; Griffiths, Emily; Almond, Solomon; University of Chester; Public Health England (BioMed Central, 2016-01-24)
      Background: There were at least 31,171 metal-on-metal (MoM) hip implants in the UK between 2003 and 2011. Some of these were subject to failure and widescale recalls and revisions followed. Method This is a presentation of ten cases (mean age 60 years) where we evaluated neuropsychiatric morbidity following metal-on-metal hip implant failure and revision. Implants were ASR total hip replacement (acetabular implant, taper sleeve adaptor and unipolar femoral implants) performed between 2005 and 2009. This case series describes, for the first time, neuropsychiatric complications after revision where there has been cobalt and chromium toxicity. Results Pre-revision surgery, nine patients had toxic levels of chromium and cobalt (mean level chromium 338 nmol/l, mean cobalt 669.4 nmol/l). Depression assessment showed 9 of 9 respondents fulfilled the BDI criteria for depression and 3 of these were being treated. 7 of 9 patients showing short term memory deficit with mean mini mental state examination score of 24.2. The normal population mean MMSE for this group would be expected to be 28 with <25 indicating possible dementia. Conclusions We found neurocognitive and depressive deficits after cobalt and chromium metallosis following MoM implant failure. Larger studies of neurocognitive effects are indicated in this group. There may be implications for public health.
    • Nicotinamide alone accelerates the conversion of mouse embryonic stem cells into mature neuronal populations.

      Griffin, Sile M.; Pickard, Mark R.; Orme, Rowan P.; Hawkins, Clive P.; Williams, Adrian C.; Fricker, Rosemary; Keele University; University of Chester; University Hospital of North Staffordshire; University of Birmingham (Public Library of Science, 2017-08-17)
      Vitamin B3 has been shown to play an important role during embryogenesis. Specifically, there is growing evidence that nicotinamide, the biologically active form of vitamin B3, plays a critical role as a morphogen in the differentiation of stem cells to mature cell phenotypes, including those of the central nervous system (CNS). Detailed knowledge of the action of small molecules during neuronal differentiation is not only critical for uncovering mechanisms underlying lineage-specification, but also to establish more effective differentiation protocols to obtain clinically relevant cells for regenerative therapies for neurodegenerative conditions such as Huntington's disease (HD). Thus, this study aimed to investigate the potential of nicotinamide to promote the conversion of stem cells to mature CNS neurons. METHODS: Nicotinamide was applied to differentiating mouse embryonic stem cells (mESC; Sox1GFP knock-in 46C cell line) during their conversion towards a neural fate. Cells were assessed for changes in their proliferation, differentiation and maturation; using immunocytochemistry and morphometric analysis methods. RESULTS: Results presented indicate that 10 mM nicotinamide, when added at the initial stages of differentiation, promoted accelerated progression of ESCs to a neural lineage in adherent monolayer cultures. By 14 days in vitro (DIV), early exposure to nicotinamide was shown to increase the numbers of differentiated βIII-tubulin-positive neurons. Nicotinamide decreased the proportion of pluripotent stem cells, concomitantly increasing numbers of neural progenitors at 4 DIV. These progenitors then underwent rapid conversion to neurons, observed by a reduction in Sox 1 expression and decreased numbers of neural progenitors in the cultures at 14 DIV. Furthermore, GABAergic neurons generated in the presence of nicotinamide showed increased maturity and complexity of neurites at 14 DIV. Therefore, addition of nicotinamide alone caused an accelerated passage of pluripotent cells through lineage specification and further to non-dividing mature neurons. CONCLUSIONS: Our results show that, within an optimal dose range, nicotinamide is able to singly and selectively direct the conversion of embryonic stem cells to mature neurons, and therefore may be a critical factor for normal brain development, thus supporting previous evidence of the fundamental role of vitamins and their metabolites during early CNS development. In addition, nicotinamide may offer a simple effective supplement to enhance the conversion of stem cells to clinically relevant neurons.
    • Novel anti-oxidant properties of cobalamin

      Williams, John H. H.; Andrew, Sarah M.; Altaie, Ala (University of Liverpool (University of Chester), 2009-09)
      Oxidative stress has been associated with a wide range of diseases, including cardiovascular diseases, Alzheimer's disease, atherosclerosis, Parkinson's disease and cancer. It also plays a role in the ageing process. Hyperhomocysteimia is commonly found to be associated with these diseases. The hyperhomocysteimia is a result of a deficiency in both folate and cobalamin Folate is known to reduce Hey and protect cells from apoptosis, but there are no studies investigating the impact of cobalamin on apoptosis induced by oxidative stress or the mechanism(s) of the protection. The aims of the research are to investigate the protective role of cobalamin and the possible mechanism(s) for this protection. It also examines the protective role of novel cobalamin and investigates their superior protection. The methods used in this research for apoptosis detection we used caspase-3 and the annexin-V, while for necrosis we used PI staining, where cell viability were detected using MTS assay. We also measured the generation of superoxide by Lucigenin-enhanced chemiluminescence and reactive oxygene species by using the redox active prob DCFH-DA. Moreover, the intracellular proteins were measured via staining with specific fluorescent-conjugated antibodies were detected using flowcytometry. Our result demonstrated that 25|iM of cobalamin protects cells from apoptosis. The protection by cobalamin was associated with induction of iHsp72 and iHO-1, and these are shown to be essential for the protection. Furthermore, our research demonstrated a novel mechanism of cobalamin-apoptosis protection involving induction of NfkB, ERK1/2 and AKT signal transduction pathways. In order to protect cells from apoptosis induced by oxidative stress, cobalamin induces the pNfkB which in turn regulate the iNOS and HO-1 induction. Cobalamin also induces thepERK1/2 which regulates the induction of Hps72 and Nrf2. And finally, pAKT induced by cobalamin which regulate the Nrf2 and HO-1 induction. The inhibition of any of theses pathways leads to loss the protection. The GSCbl and NACCbl provide a superior protection against oxidative stress, this protection involved induction of the signal transduction pathways and Hsps. To conclude; cobalamin provides protection against cells death induced by oxidative stress. Cobalamin achieves this by multiple pathways which include direct antioxidant stimulation and induction of signal transduction pathways. Different cobalamin derivatives have superior protections. These finding are a useful pharmaceutical tool in the treatment of the oxidative stress related diseases.
    • Nurses attitudes and beliefs to attempted suicide in Southern India

      Jones, Steven; Krishna, Murali; Rajendra, Raj G.; Keenan, Paul; University of Chester (Taylor and Francis, 2015-05-20)
      Background: There is growing global interest into the attitudes and clinical management of persons who have attempted suicide. Aims: The principal purpose was to determine senior nursing staff attitudes towards patients who had attempted suicide from a professional and cultural perspective, which might influence care following hospital admission. The focus concerned nursing staff interactions at a psychological level that compete with physical tasks on general hospital wards. Methods: A qualitative methodology was employed with audio-taped interviews utilising four level data coding. This article reports on a group of 15 nursing staff from a large general hospital in Mysore, Southern India. Results: Findings suggested that patient care and treatment is directly influenced by the nurse’s religious beliefs within a general hospital setting with physical duties prioritised over psychological support, which was underdeveloped throughout the participant group. Conclusion: The results allow a series of recommendations for educational and skills initiatives before progressing to patient assessment and treatment projects and cross-cultural comparison studies. In addition, interventions must focus on current resources and context to move the evidence-based suicide prevention forward.
    • Oesophageal stenting: Status quo and future challenges.

      Kaltsidis, Harry; Mansoor, Wasat; Park, Jung-Hoon; Song, Ho-Young; Edwards, Derek W.; Laasch, Hans-Ulrich (2018-06-11)
      Oesophageal stents are widely used for palliating dysphagia from malignant obstruction. They are also used with increasing frequency in the treatment of oesophageal perforation, as well as benign strictures from a variety of causes. Improved oncological treatments have led to prolonged survival of patients treated with palliative intent; as a consequence, stents need to function and last longer in order to avoid repeat procedures. There is also increasing need for meticulous procedure planning, careful selection of the device most appropriate for the individual patient and planned follow-up. Furthermore, as more patients are cured, there will be more issues with resultant long-term side-effects, such as recalcitrant strictures due to radiotherapy or anastomotic scarring, which will have to be addressed. Stent design needs to keep up with the progress of cancer treatment, in order to offer patients the best possible long-term result. This review article attempts to illustrate the changing realities in oesophageal stenting, differences in current stent designs and behaviour, as well as the pressing need to refine and modify devices in order to meet the new challenges.
    • Office workers’ experiences of attempts to reduce sitting-time: An exploratory, mixed- methods uncontrolled intervention pilot study

      Dewitt, Stephen; Hall, Jennifer; Smith, Lee; Buckley, John P.; Biddle, Stuart J. H.; Mansfield, Louise; Gardner, Benjamin; University of Chester (BMC Springer Nature, 2019-06-25)
      Background: Office workers typically sit for most of the workday, which has been linked to physical and mental ill- health and premature death. This mixed-methods study sought to identify barriers and facilitators to reducing sitting and increasing standing among office workers who received an intervention prototype (the ‘ReSiT [Reducing Sitting Time] Study’). The intervention comprised a sit-stand workstation and tailored advice to enhance motivation, capability and opportunity to displace sitting with standing. Methods: Twenty-nine UK university office workers (aged ≥18y, working ≥3 days per week, most time spent at a seated desk) participated in a 13-week uncontrolled study. They were initially monitored for one-week. In a subsequent face-to-face consultation, participants received sitting time feedback from a prior one-week monitoring period, and selected from a set of tailored sitting-reduction techniques. Quantitative data comprising sitting, standing and stepping time, which were objectively monitored for 7 consecutive days across three post- intervention timepoints, were descriptively analysed. Qualitative data, from semi-structured interviews conducted at 1, 6 and 12-weeks post-intervention, were thematically analysed. Results: Compared to baseline, mean sitting time decreased at weeks 1, 6 and 12 by 49.7mins, 118.2mins, and 109.7mins respectively. Despite prior concerns about colleagues’ reactions to standing, many reported encouragement from others, and standing could be equally conducive to social interaction or creating private, personal space. Some perceived less cognitively-demanding tasks to be more conducive to standing, though some found standing offered a valued break from challenging tasks. Participants prioritised workload over sitting reduction and were more likely to stand after rather than during work task completion. Temporary context changes, such as holidays, threatened to derail newfound routines. Conclusions: Our findings emphasise the importance of understanding workers’ mental representations of their work, and the social functions of sitting and standing in the workplace. Workplace intervention developers should incorporate a pre-intervention sitting time monitoring period, encourage workers to identify personally meaningful tasks and cues for standing, and build organisational support for sitting-reduction. We will use these insights to refine our intervention for self-administered delivery. Trial registration: ISRCTN29395780 (registered 21 November 2016). Keywords: Sedentary behaviour, Workplace, Qualitative, Occupational health