Oxidant and Carbonyl Stress-Related Apoptosis in End-Stage Kidney Disease: Impact of Membrane Flux

Apoptosis is a highly regulated process which mostly affects cell-mediated immunity. In this open-label, randomized, prospective clinical study, we determined the impact in 10 hemodialysis patients treated with high-, medium-, and low-flux membranes on spontaneous or plasma-induced apoptosis, on monocytes, as well as on oxidant and carbonyl stress. High- and medium-flux membranes significantly reduced patients' plasma-dependent proapoptotic activity on U937 monocytic cell lines. Patients who had the highest levels of plasma-induced proapoptotic activity exhibited the highest plasma levels of advanced oxidation protein products (AOPPs) and carbonyls. Plasma carbonyl residues but not AOPPs were significantly lowered. Finally, a significant correlation could be drawn between the extent of plasma-induced proapoptotic activity and both plasma carbonyl and AOPP levels

A pilot study of brisk walking in sedentary combination antiretroviral treatment (cART)- treated patients: benefit on soluble and cell inflammatory markers

Background: Chronic HIV infection is associated with low-level inflammation and increased risk of chronic diseases and mortality. The objective was to assess the effects of moderate intensity exercise on metabolic and inflammatory markers in HIV-infected treated persons. Methods: This was a pilot study enrolling cART-treated, sedentary persons with metabolic complications in a 12-week protocol, consisting of three sessions per week of 60 min brisk walking with (strength-walk group) or without (walk group) 30 min circuit-training. Assessments at baseline and week 12 (W12) included body morphometrics and total body dual-energy X-ray absorptiometry; lipid and glucose blood profile; plasma level of high sensitivity C-reactive protein (hsCRP), interleukin-6 (IL-6), D-dimer, interleukin-18 (IL-18), soluble CD14, and CD38 and HLA-DR expression on CD4+ and CD8+ T-cells. Results: Forty-nine patients were included and 35 (71%) completed the program: 21 in the walk and 14 in the strength-walk group. At W12, significant improvements were observed of body mass index, waist and hip circumference, and total cholesterol both overall and in the walk group, and of LDL cholesterol in both training groups. In the whole group, significant reductions were observed in hsCRP, IL-6, D-dimer, IL-18, and of CD8 +/CD38+/HLA-DR+ cell frequencies. HsCRP and CD8+/CD38+/HLA-DR+ frequency decreased significantly in both training groups when examined separately whereas IL-6 and D-dimer in the walk group only. Conclusions: Brisk walking, with or without strength exercise, could improve lipid profile and inflammatory markers in chronic HIV infection

Co-Graft of Allogeneic Immune Regulatory Neural Stem Cells (NPC) and Pancreatic Islets Mediates Tolerance, while Inducing NPC-Derived Tumors in Mice

Data available on the immunomodulatory properties of neural stem/precursor cells (NPC) support their possible use as modulators for immune-mediated process. The aim of this study was to define whether NPC administered in combination with pancreatic islets prevents rejection in a fully mismatched allograft model.Diabetic Balb/c mice were co-transplanted under the kidney capsule with pancreatic islets and GFP(+) NPC from fully mismatched C57BL/6 mice. The following 4 groups of recipients were used: mice receiving islets alone; mice receiving islets alone and treated with standard immunosuppression (IL-2Ralpha chain mAbs + FK506 + Rapamycin); mice receiving a mixed islet/NPC graft under the same kidney capsule (Co-NPC-Tx); mice receiving the islet graft under the left kidney capsule and the NPC graft under the right kidney capsule (NPC-Tx). Our results demonstrate that only the co-transplantation and co-localization of NPC and islets (Co-NPC-Tx) induce stable long-term graft function in the absence of immunosuppression. This condition is associated with an expansion of CD4(+)CD25(+)FoxP3(+) T regulatory cells in the spleen. Unfortunately, stable graft function was accompanied by constant and reproducible development of NPC-derived cancer mainly sustained by insulin secretion.These data demonstrate that the use of NPC in combination with islets prevents graft rejection in a fully mismatched model. However, the development of NPC-derived cancer raises serious doubts about the safety of using adult stem cells in combination with insulin-producing cells outside the original microenvironment

Activation and nuclear translocation of PKCε promotes skeletal muscle cell differentiation via HMGA1 downregulation

The role of novel PKCs in skeletal muscle differentiation has recently emerged. PKCθ is the most expressed isoform of PKCs in muscle and it promotes the fusion of myoblasts [1]. Recently, we have demonstrated that PKCε is implicated in myocardiocyte differentiation of bone marrow mesenchymal stem cells [2] but the role of PKCε in skeletal muscle cell regeneration has only recently emerged [3]. We here demonstrate that both nuclear and cytoplasmic fractions of PKCε are up-regulated during in vitro C2C12 cell line and satellite cell differentiation. We also show that PKCε is able to modulate myogenic differentiation genes via a downmodulation of HMGA1 proteins that promotes myogenin accumulation and mature myoblast formation. To study the effects of PKCε on muscle regeneration, we have used the in vivo model of CTX-induced skeletal muscle injury. We show that the upregulation of PKCε also occurs in vivo, particularly in the centro-nucleated regenerating fibers that are derived from the fusion process of the resident satellite cells, suggesting a role for PKCε in human satellite cell-driven muscle repair and substitution, with clinically relevant implications in human muscle pathology

Joint mobility/muscular chain elasticity and motor coordination in a cohort of 9-11 years school children exposed to specifically designed and professionally guided training

Beside the positive role that an active lifestyle plays in the physical and emotional well-being of a child, physically active children have lower risks to encounter injury as adults. However, many groups have reported that only a small population of children in western countries are sufficiently active (1, 2). The aim of this study was to investigate whether joint mobility/muscular elasticity and coordination were related to a merely active lifestyle or could be significantly improved in the presence of a collective, easy-to-perform, but specifically-designed and professionally-guided school program. Specific functional and anthropometric parameters were single-blind tested on 277 children (aged 9-11 years). 148 were randomly assigned to a school-based physical education program specifically designed to increase coordination and elasticity and supervised by professionals, while 129 (control group) continued their usual physical activity at school, with no specific program. The specific program generated a significant improvement of joint mobility and coordination abilities as compared to non-specific physical activity. As a secondary end-point, gender and BMI-related differences emerged during the study, showing that females respond better to a low intensity program, while males benefit of a higher intensity (or a differently designed) program, particularly when belonging to overweight/ obese BMI classes. These results, building up on those from our and other groups, should orient decision-makers in the area of physical exercise for primary school children in favour of specifically designed programs based on demographic and anthropometric data

Gender-related differences in the prevalence of voice disorders and awareness of dysphonia

Objective: Considering the impact of dysphonia on public health and the increasing attention to patient-centred care, we evaluated sex-related differences in the prevalence of benign voice disorders, awareness of dysphonia and voice therapy (VT) results. Methods: One hundred and seventy-one patients, 129 females and 42 males, with functional or organic benign dysphonia underwent Voice Handicap Index (VHI), auditory-perceptual dysphonia severity scoring (GRBAS) and acoustic analysis (Jitter%, Shimmer%, NHR) before and after VT. Results: Prevalence of each voice disorder was significantly higher among females. Mean time-to-diagnosis (time elapsed until medical consultation) was not different between males and females. The refusal of therapy and VT adherence (mean number of absences and premature dropout) were similar in the two groups. Pre-VT VHI and "G" parameter were worse in women. The percentage of women with abnormal acoustic analysis was significantly higher. Post-VT VHI gain was higher in women, whereas "G" parameter improvement did not differ by sex. Conclusions: Our study showed a higher prevalence of voice disorders in females. Awareness of dysphonia was not gender related. Females started with worse voice subjective perception and acoustic analysis, but they perceived greater improvement after therapy

New Algorithm to Determine True Colocalization in Combination with Image Restoration and Time-Lapse Confocal Microscopy to Map Kinases in Mitochondria

The subcellular localization and physiological functions of biomolecules are closely related and thus it is crucial to precisely determine the distribution of different molecules inside the intracellular structures. This is frequently accomplished by fluorescence microscopy with well-characterized markers and posterior evaluation of the signal colocalization. Rigorous study of colocalization requires statistical analysis of the data, albeit yet no single technique has been established as a standard method. Indeed, the few methods currently available are only accurate in images with particular characteristics. Here, we introduce a new algorithm to automatically obtain the true colocalization between images that is suitable for a wide variety of biological situations. To proceed, the algorithm contemplates the individual contribution of each pixel's fluorescence intensity in a pair of images to the overall Pearsońs correlation and Manders' overlap coefficients. The accuracy and reliability of the algorithm was validated on both simulated and real images that reflected the characteristics of a range of biological samples. We used this algorithm in combination with image restoration by deconvolution and time-lapse confocal microscopy to address the localization of MEK1 in the mitochondria of different cell lines. Appraising the previously described behavior of Akt1 corroborated the reliability of the combined use of these techniques. Together, the present work provides a novel statistical approach to accurately and reliably determine the colocalization in a variety of biological images

Managing wolf impacts on sheep husbandry: a collaborative implementation and assessment of damage prevention measures in an agricultural landscape

Wolves in Europe are expanding their range and significantly impacting farming livelihoods and landscapes. Damage prevention measures such as livestock guarding dogs and night-time enclosures have proven successful in mitigating losses. However, they are often implemented as top-down measures without a proper understanding of the farming dynamics they are meant to alter, making them unappealing and difficult to implement for farmers. Semi-extensive, small scale livestock farming systems are particularly vulnerable and diverse, requiring specific care and catered support when addressing issues related to wildlife management. In these contexts, it is crucial to employ adaptive management approaches that enable solutions to be collaboratively designed at the grassroots level. Here we propose a method for centring the experiences and knowledge of local farmers to co-produce damage prevention practices that better address their needs. We developed this approach in the course of the LIFE MEDWOLF project, which was implemented in the province of Grosseto, Italy, between 2012 and 2017. The project brought together local authorities, environmental associations, farming unions and individual farmers to develop tailor-made damage prevention measures and assess their technical and economic impact, through a stepwise process. Collaboration with 86 local farmers resulted in >50 modifications to the original project plan, and an overall 50% reduction of preyed livestock in farms that participated in the project. Our findings highlight the benefits of collaboratively designing, implementing, and monitoring damage prevention measures with farmers. Based on these results, we reflect on the importance of integrating local and scientific knowledge, on the implications this has had on stakeholder relations, and on the challenges that we faced in upscaling this management approach

Isolation and characterization of non-O157 Shiga toxin-producing <i>Escherichia coli</i> from beef carcasses, cuts and trimmings of abattoirs in Argentina

Several foods contaminated with Shiga toxin-producing Escherichia coli (STEC) are associated with human diseases. Some countries have established microbiological criteria for non-O157 STEC, thus, the absence of serogroups O26, O45, O103, O104, O111, O121, and O145 in sprouts from the European Union or ground beef and beef trimmings from the United States is mandatory. While in Argentina screening for O26, O103, O111, O145 and O121 in ground beef, ready-to-eat food, sausages and vegetables is mandatory, other countries have zero-tolerance for all STEC in chilled beef. The aim of this study was to provide data on the prevalence of non-O157 STEC isolated from beef processed in eight Argentinean cattle slaughterhouses producing beef for export and local markets, and to know the non-O157 STEC profiles through strain characterization and genotypic analysis. Samples (n = 15,965) from 3,205 beef carcasses, 9,570 cuts and 3,190 trimmings collected between March and September 2014 were processed in pools of five samples each. Pools of samples (n = 3,193) from 641 carcasses, 1,914 cuts and 638 trimming were analyzed for non-O157 STEC isolation according to ISO/CEN 13136:2012. Of these, 37 pools of carcasses (5.8%), 111 pools of cuts (5.8%) and 45 pools of trimmings (7.0%) were positive for non-O157 STEC. STEC strains (n = 200) were isolated from 193 pools of samples. The most prevalent serotypes were O174:H21, O185:H7, O8:H19, O178:H19 and O130:H11, and the most prevalent genotypes were stx2c(vh-b) and stx2a/saa/ehxA. O103:H21 strain was eae-positive and one O178:H19 strain was aggR/aaiC-positive. The prevalence of non-O157 STEC in beef carcasses reported here was low. None of the non-O157 STEC strains isolated corresponded to the non-O157 STEC serotypes and virulence profiles isolated from human cases in Argentina in the same study period. The application of microbiological criteria for each foodstuff should be determined by risk analysis in order to have a stringent monitoring system. Likewise, zero-tolerance intervention measures should be applied in beef, together with GMP and HACCP. Further, collaborative efforts for risk assessment, management and communication are extremely important to improve the safety of foodstuffs.Instituto de Genética VeterinariaFacultad de Ciencias Veterinaria