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Authors: Anna Höfer, Johannes Herrmann, Patrick Meybohm, Christopher Lotz

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Mitochondria are pivotal for energy supply and regulation of cellular functions. Deficiencies of mitochondrial metabolism have been implicated in diverse stressful conditions including infections. Platelets are key mediators for thrombo-inflammation during development and resolution of acute respiratory distress syndrome (ARDS). Previous data point to an exhausted platelet phenotype in critically-ill patients with coronavirus 19 disease (COVID-19) impacting the course of disease. The objective of this work was to characterize platelet mitochondrial metabolism in patients suffering from COVID-19 ARDSA longitudinal analysis of platelet mitochondrial metabolism in 24 patients with COVID-19 induced ARDS compared to 35 healthy controls (ctrl) was performed. Blood samples were analyzed at two time points (t1=day 1; t2=day 5-7 after study inclusion). The activity of mitochondrial citrate synthase was photometrically measured. The impact of oxidative stress on mitochondrial permeability was assessed by a photometric calcium-induced swelling assay and the activity of superoxide dismutase (SOD) by a SOD assay kit. The amount of protein carbonylation and the activity of mitochondria complexes I-IV were photometrically determined. Levels of interleukins (IL)-1α, IL-1β and tumor necrosis factor (TNF-) α were measured by a Multiplex assay kit. Median age was 54 years, 63 % were male and BMI was 29.8 kg/m2. SOFA (12; IQR: 10-15) and APACHE II (27; IQR: 24-30) indicated critical illness. Median Murray Score was 3.4 (IQR: 2.8-3.4), 21/24 (88%) required mechanical ventilation and V-V ECMO support in 14/24 (58%). Platelet counts in ARDS did not change during ICU stay (t1: 212 vs. t2: 209 x109/L). However, mean platelet volume (MPV) significantly increased (t1: 10.6 vs. t2: 11.9 fL; p<0.0001). Citrate synthase activity showed no significant differences between ctrl and ARDS patients. Calcium induced swelling was more pronounced in patients at t1 compared to t2 and to ctrl (50µM; t1: 0.006 vs. ctrl: 0.016 ΔOD; p=0.001). The amount of protein carbonylation as marker for irreversible proteomic modification constantly increased during ICU stay and compared to ctrl., without reaching significance. In parallel, superoxid dismutase activity gradually declined during ICU treatment vs. ctrl (t2: - 29 vs. ctrl.: - 17 %; p=0.0464). Complex I analysis revealed significantly stronger activity in ARDS vs. ctrl. (t1: 0.633 vs. ctrl.: 0.415 ΔOD; p=0.0086). There were no significant differences in complex II, III or IV activity in platelets from ARDS patients compared to ctrl. IL-18 constantly increased during the observation period without reaching significance. IL-1α and TNF-α did not differ from ctrl. However, IL-1β levels were significantly elevated in ARDS (t1: 16.8; t2: 16.6 vs. ctrl.: 12.4 pg/mL; p1=0.0335, p2=0.0032). This study reveals new insights in platelet mitochondrial metabolism during COVID-19 caused ARDS. it data point towards enhanced platelet activity with a pronounced turnover rate. We found increased activity of mitochondria complex I and evidence for enhanced oxidative stress. In parallel, protective mechanisms against oxidative stress were narrowed with elevated levels of IL-1β likely causing a pro-apoptotic environment. These mechanisms may contribute to platelet exhaustion in ARDS.

Keywords: acute respiratory distress syndrome (ARDS), coronavirus 19 disease (COVID-19), oxidative stress, platelet mitochondrial metabolism

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Authors: Lei Ye, Min Liu, Yong-Li Gao, Jun Zhang

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Purpose: The aims of the study are to investigate the status of health-promoting lifestyle and to compare the healthy lifestyle of emergency nurses in different levels of hospitals in Sichuan province, China. The investigation is mainly about the health-promoting lifestyle, including spiritual growth, health responsibility, physical activity, nutrition, interpersonal relations, stress management. Then the factors were analyzed influencing the health-promoting lifestyle of emergency nurses in hospitals of Sichuan province in order to find the relevant models to provide reference evidence for intervention. Study Design: A cross-sectional research method was adopted. Stratified cluster sampling, based on geographical location, was used to select the health facilities of 1389 emergency nurses in 54 hospitals from Sichuan province in China. Method: The 52-item, six-factor structure Health-Promoting Lifestyle Profile II (HPLP- II) instrument was used to explore participants’ self-reported health-promoting behaviors and measure the dimensions of health responsibility, physical activity, nutrition, interpersonal relations, spiritual growth, and stress management. Demographic characteristics, education, work duration, emergency nursing work duration and self-rated health status were documented. Analysis: Data were analyzed through SPSS software ver. 17.0. Frequency, percentage, mean ± standard deviation were used to describe the general information, while the Nonparametric Test was used to compare the constituent ratio of general data of different hospitals. One-way ANOVA was used to compare the scores of health-promoting lifestyle in different levels hospital. A multiple linear regression model was established. P values which were less than 0.05 determined statistical significance in all analyses. Result: The survey showed that the total score of health-promoting lifestyle of nurses at emergency departments in Sichuan Province was 120.49 ± 21.280. The relevant dimensions are ranked by scores in descending order: interpersonal relations, nutrition, health responsibility, physical activity, stress management, spiritual growth. The total scores of the three-A hospital were the highest (121.63 ± 0.724), followed by the senior class hospital (119.7 ± 1.362) and three-B hospital (117.80 ± 1.255). The difference was statistically significant (P=0.024). The general data of nurses was used as the independent variable which includes age, gender, marital status, living conditions, nursing income, hospital level, Length of Service in nursing, Length of Service in emergency, Professional Title, education background, and the average number of night shifts. The total score of health-promoting lifestyle was used as dependent variable; Multiple linear regression analysis method was adopted to establish the regression model. The regression equation F = 20.728, R2 = 0.061, P < 0.05, the age, gender, nursing income, turnover intention and status of coping stress affect the health-promoting lifestyle of nurses in emergency department, the result was statistically significant (P < 0.05 ). Conclusion: The results of the investigation indicate that it will help to develop health promoting interventions for emergency nurses in all levels of hospital in Sichuan Province through further research. Managers need to pay more attention to emergency nurses’ exercise, stress management, self-realization, and conduct intervention in nurse training programs.

Keywords: emergency nurse, health-promoting lifestyle profile II, health behaviors, lifestyle

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Authors: Rebecca Laycock

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Many Urban Agriculture (UA) initiatives, including community/allotment gardens, Community Supported Agriculture, and community/social farms, depend on volunteers. However, initiatives supported or run by volunteers are often faced with a high turnover of labour as a result of the involvement of episodic volunteers (a term describing ad hoc, one-time, and seasonal volunteers), leading to challenges with maintaining project continuity and retaining skills/knowledge within the initiative. This is a notable challenge given that food growing is a knowledge intensive activity where the fruits of labour appear months or sometimes years after investment. Participatory Action Research (PAR) is increasingly advocated for in the field of UA as a solution-oriented approach to research, providing concrete results in addition to advancing theory. PAR is a cyclical methodological approach involving researchers and stakeholders collaboratively 'identifying' and 'theorising' an issue, 'planning' an action to address said issue, 'taking action', and 'reflecting' on the process. Through iterative cycles and prolonged engagement, the theory is developed and actions become better tailored to the issue. The demand for PAR in UA research means that understanding how to use PAR with episodic volunteers is of critical importance. The aim of this paper is to explore (1) the challenges of doing PAR in UA initiatives with episodic volunteers, and (2) how PAR can be harnessed to advance sustainable development of UA through theoretically-informed action. A 2.5 year qualitative PAR study on three English case study student-led food growing initiatives took place between 2014 and 2016. University UA initiatives were chosen as exemplars because most of their volunteers were episodic. Data were collected through 13 interviews, 6 workshops, and a research diary. The results were thematically analysed through eclectic coding using Computer-Assisted Qualitative Data Analysis Software (NVivo). It was found that the challenges of doing PAR with transient participants were (1) a superficial understanding of issues by volunteers because of short term engagement, resulting in difficulties ‘identifying’/‘theorising’ issues to research; (2) difficulties implementing ‘actions’ given those involved in the ‘planning’ phase often left by the ‘action’ phase; (3) a lack of capacity of participants to engage in research given the ongoing challenge of maintaining participation; and (4) that the introduction of the researcher acted as an ‘intervention’. The involvement of a long-term stakeholder (the researcher) changed the group dynamics, prompted critical reflections that had not previously taken place, and improved continuity. This posed challenges for providing a genuine understanding the episodic volunteering PAR initiatives, and also challenged the notion of what constitutes an ‘intervention’ or ‘action’ in PAR. It is recommended that researchers working with episodic volunteers using PAR should (1) adopt a first-person approach by inquiring into the researcher’s own experience to enable depth in theoretical analysis to manage the potentially superficial understandings by short-term participants; and (2) establish safety mechanisms to address the potential for the research to impose artificial project continuity and knowledge retention that will end when the research does. Through these means, we can more effectively use PAR to conduct solution-oriented research about UA.

Keywords: community garden, continuity, first-person research, higher education, knowledge retention, project management, transience, university

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Authors: Zuzana Tatarkova, Ivana Pilchova, Michal Cibulka, Martin Kolisek, Peter Racay, Peter Kaplan

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Introduction: Normal functioning of mitochondria is crucial for cardiac performance. Mitochondria undergo mitophagy and biogenesis, and mitochondrial proteins are subject to extensive post-translational modifications. The state of mitochondrial homeostasis reflects overall cellular fitness and longevity. Perturbed mitochondria produce less ATP, release greater amounts of reactive molecules, and are more prone to apoptosis. Therefore mitochondrial turnover is an integral aspect of quality control in which dysfunctional mitochondria are selectively eliminated through mitophagy. Currently, the progressive deterioration of physiological functions is seen as accumulation of modified/damaged proteins with limiting regenerative ability and disturbance of such affected protein-protein communication throughout aging in myocardial cells. Methodologies: For our study was used immunohistochemistry, biochemical methods: spectrophotometry, western blotting, immunodetection as well as more sophisticated 2D electrophoresis and mass spectrometry for evaluation protein-protein interactions and specific post-translational modification. Results and Discussion: Mitochondrial stress response to reactive species was evaluated as electron transport chain (ETC) complexes, redox-active molecules, and their possible communication. Protein-protein interactions revealed a strong linkage between age and ETC protein subunits. Redox state was strongly affected in senescent mitochondria with shift in favor of more pro-oxidizing condition within cardiomyocytes. Acute myocardial ischemia and ischemia-reperfusion (IR) injury affected ETC complexes I, II and IV with no change in complex III. Ischemia induced decrease in total antioxidant capacity, MnSOD, GSH and catalase activity with recovery in some extent during reperfusion. While MnSOD protein content was higher in IR group, activity returned to 95% of control. Nitric oxide is one of the biological molecules that can out compete MnSOD for superoxide and produce peroxynitrite. This process is faster than dismutation and led to the 10-fold higher production of nitrotyrosine after IR injury in adult with higher protection in senescent ones. 2D protein profiling revealed 140 mitochondrial proteins, 12 of them with significant changes after IR injury and 36 individual nitrotyrosine-modified proteins further identified by mass spectrometry. Linking these two groups, 5 proteins were altered after IR as well as nitrated, but only one showed massive nitration per lowering content of protein after IR injury in adult. Conclusions: Senescent cells have greater proportion of protein content, which might be modulated by several post-translational modifications. If these protein modifications are connected to functional consequences and protein-protein interactions are revealed, link may lead to the solution. Assume all together, dysfunctional proteostasis can play a causative role and restoration of protein homeostasis machinery is protective against aging and possibly age-related disorders. This work was supported by the project VEGA 1/0018/18 and by project 'Competence Center for Research and Development in the field of Diagnostics and Therapy of Oncological diseases', ITMS: 26220220153, co-financed from EU sources.

Keywords: aging heart, mitochondria, proteomics, redox state

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Authors: Narottam Puri, Gurvinder Kaur

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Background: The National Accreditation Board for Hospitals & Healthcare Providers (NABH) is India’s apex standard setting accrediting body in health care which evaluates and accredits healthcare organizations. NABH requires accredited organizations to become reaccredited every three years. It is often though that once the initial accreditation is complete, the foundation is set and reaccreditation is a much simpler process. Fortis Hospital, Shalimar Bagh, a part of the Fortis Healthcare group is a 262 bed, multi-specialty tertiary care hospital. The hospital was successfully accredited in the year 2012. On completion of its first cycle, the hospital underwent a reaccreditation assessment in the year 2015. This paper aims to gain a better understanding of the challenges that accredited hospitals face when preparing for a renewal of their accreditations. Methods: The study was conducted using a cross-sectional mixed methods approach; semi-structured interviews were conducted with senior leadership team and staff members including doctors and nurses. Documents collated by the QA team while preparing for the re-assessment like the data on quality indicators: the method of collection, analysis, trending, continual incremental improvements made over time, minutes of the meetings, amendments made to the existing policies and new policies drafted was reviewed to understand the challenges. Results: The senior leadership had a concern about the cost of accreditation and its impact on the quality of health care services considering the staff effort and time consumed it. The management was however in favor of continuing with the accreditation since it offered competitive advantage, strengthened community confidence besides better pay rates from the payors. The clinicians regarded it as an increased non-clinical workload. Doctors felt accountable within a professional framework, to themselves, the patient and family, their peers and to their profession; but not to accreditation bodies and raised concerns on how the quality indicators were measured. The departmental leaders had a positive perception of accreditation. They agreed that it ensured high standards of care and improved management of their functional areas. However, they were reluctant in sparing people for the QA activities due to staffing issues. With staff turnover, a lot of work was lost as sticky knowledge and had to be redone. Listing the continual quality improvement initiatives over the last 3 years was a challenge in itself. Conclusion: The success of any quality assurance reaccreditation program depends almost entirely on the commitment and interest of the administrators, nurses, paramedical staff, and clinicians. The leader of the Quality Movement is critical in propelling and building momentum. Leaders need to recognize skepticism and resistance and consider ways in which staff can become positively engaged. Involvement of all the functional owners is the start point towards building ownership and accountability for standards compliance. Creativity plays a very valuable role. Communication by Mail Series, WhatsApp groups, Quizzes, Events, and any and every form helps. Leaders must be able to generate interest and commitment without burdening clinical and administrative staff with an activity they neither understand nor believe in.

Keywords: NABH, reaccreditation, quality assurance, quality indicators

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Authors: Bassey O. Bassey

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Benchmarking has progressively become entrenched as a requisite activity for process improvement and enhancing service delivery at petroleum jetties and terminals, most especially during tanker discharge operations at the ship – shore interface, as avoidable delays result in extra operating costs, non-productive time, high demurrage payments and ultimate product scarcity. The jetty and terminal in focus had been operational for 3 and 8 years respectively, with proper operational and logistic records maintained to evaluate their progress over time in order to plan and implement modifications and review of procedures for greater technical and economic efficiency. Regular and emergency staff meetings were held on a team, departmental and company-wide basis to progressively address major challenges that were encountered during each operation. The process and outcome of the resultant collectively planned changes carried out within the past two years forms the basis of this paper, which mirrors the initiatives effected to enhance operational and maintenance excellence at the affected facilities. Operational modifications included a second cargo receipt line designated for gasoline, product loss control at jetty and shore ends, enhanced product recovery and quality control, and revival of terminal–jetty backloading operations. Logistic improvements were the incorporation of an internal logistics firm and shipping agency, fast tracking of discharge procedures for tankers, optimization of tank vessel selection process, and third party product receipt and throughput. Maintenance excellence was achieved through construction of two new lay barges and refurbishment of the existing one; revamping of existing booster pump and purchasing of a modern one as reserve capacity; extension of Phase 1 of the jetty to accommodate two vessels and construction of Phase 2 for two more vessels; regular inspection, draining, drying and replacement of cargo hoses; corrosion management program for all process facilities; and an improved, properly planned and documented maintenance culture. Safety, environmental and security compliance were enhanced by installing state-of-the-art fire fighting facilities and equipment, seawater intake line construction as backup for borehole at the terminal, remediation of the shoreline and marine structures, modern spill containment equipment, improved housekeeping and accident prevention practices, and installation of hi-technology security enhancements, among others. The end result has been observed over the past two years to include improved tanker turnaround time, higher turnover on product sales, consistent product availability, greater indigenous human capacity utilisation by way of direct hires and contracts, as well as customer loyalty. The lessons learnt from this exercise would, therefore, serve as a model to be adapted by other operators of similar facilities, contractors, academics and consultants in a bid to deliver greater sustainability and profitability of operations at the ship – shore interface to this strategic industry.

Keywords: benchmarking, optimisation, petroleum jetty, petroleum terminal

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Authors: Leila Noori, Rosario Barone, Francesca Rappa, Antonella Marino Gammazza, Alessandra Maria Vitale, Giuseppe Donato Mangano, Giusy Sentiero, Filippo Macaluso, Kathryn H. Myburgh, Francesco Cappello, Federica Scalia

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The neuromuscular system controls, directs, and allows movement of the body through the action of neural circuits, which include motor neurons, sensory neurons, and skeletal muscle fibers. Protein homeostasis of the involved cytotypes appears crucial to maintain the correct and prolonged functions of the neuromuscular system, and both neuronal cells and skeletal muscle fibers express significant quantities of protein chaperones, the molecular machinery responsible to maintain the protein turnover. Genetic mutations or defective post-translational modifications of molecular chaperones (i.e., genetic or acquired chaperonopathies) may lead to neuromuscular disorders called as neurochaperonopathies. The limited knowledge of the effects of the defective chaperones on skeletal muscle fibers and neurons impedes the progression of therapeutic approaches. A distinct genetic variation of CCT5 gene encoding for the subunit 5 of the chaperonin CCT (Chaperonin Containing TCP1; also known as TRiC, TCP1 Ring Complex) was recently described associated with severe distal motor neuropathy by our team. In this study, we investigated the histopathological abnormalities of the skeletal muscle biopsy of the pediatric patient affected by the mutation Leu224Val in the CCT5 subunit. We provide molecular and structural features of the diseased skeletal muscle tissue that we believe may be useful to identify undiagnosed cases of this rare genetic disorder. We investigated the histological abnormalities of the affected tissue via hematoxylin and eosin staining. Then we used immunofluorescence and qPCR techniques to explore the expression and distribution of CCT5 in diseased and healthy skeletal muscle tissue. Immunofluorescence and immunohistochemistry assays were performed to study the sarcomeric and structural proteins of skeletal muscle, including actin, myosin, tubulin, troponin-T, telethonin, and titin. We performed Western blot to examine the protein expression of CCT5 and some heat shock proteins, Hsp90, Hsp60, Hsp27, and α-B crystallin, along with the main client proteins of the CCT5, actin, and tubulin. Our findings revealed muscular atrophy, abnormal morphology, and different sizes of muscle fibers in affected tissue. The swollen nuclei and wide interfiber spaces were seen. Expression of CCT5 had been decreased and showed a different distribution pattern in the affected tissue. Altered expression, distribution, and bandage pattern were detected by confocal microscopy for the interested muscular proteins in tissue from the patient compared to the healthy control. Protein levels of the studied Hsps normally located at the Z-disk were reduced. Western blot results showed increased levels of the actin and tubulin proteins in the diseased skeletal muscle biopsy compared to healthy tissue. Chaperones must be expressed at high levels in skeletal muscle to counteract various stressors such as mechanical, oxidative, and thermal crises; therefore, it seems relevant that defects of molecular chaperones may result in damaged skeletal muscle fibers. So far, several chaperones or cochaperones involved in neuromuscular disorders have been defined. Our study shows that alteration of the CCT5 subunit is associated with the damaged structure of skeletal muscle fibers and alterations of chaperone system components and paves the way to explore possible alternative substrates of chaperonin CCT. However, further studies are underway to investigate the CCT mechanisms of action to design applicable therapeutic strategies.

Keywords: molecular chaperones, neurochaperonopathy, neuromuscular system, protein homeostasis

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