Hepatic steatosis in hepatitis C is a storage disease due to HCV interaction with microsomal triglyceride transfer protein (MTP)

Liver steatosis is a frequent histological feature in patients chronically infected with hepatitis C virus (HCV). The relationship between HCV and hepatic steatosis seems to be the result of both epigenetic and genetic factors. In vivo and in vitro studies have shown that HCV can alter intrahepatic lipid metabolism by affecting lipid synthesis, oxidative stress, lipid peroxidation, insulin resistance and the assembly and secretion of VLDL. Many studies suggest that HCV-related steatosis might be the result of a direct interaction between the virus and MTP. It has been demonstrated that MTP is critical for the secretion of HCV particles and that inhibition of its lipid transfer activity reduces HCV production. However, higher degrees of hepatic steatosis were found in chronic hepatitis C patients carrying the T allele of MTP -493G/T polymorphism that seems to be associated with increased MTP transcription. We propose here that liver steatosis in hepatitis C could be a storage disease induced by the effects of the virus and of its proteins on the intracellular lipid machinery and on MTP. Available data support the hypothesis that HCV may modulate MTP expression and activity through a number of mechanisms such as inhibition of its activity and transcriptional control. Initial up regulation could favour propagation of HCV while down regulation in chronic phase could cause impairment of triglyceride secretion and excessive lipid accumulation, with abnormal lipid droplets facilitating the "storage" of virus particles for persistent infection

reusing built heritage design for the sharing economy

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Avaliações dos níveis pré e pós-prandiais de ácidos biliares séricos e amônia plasmática em felinos sadios e o efeito de tempo de congelamento do plasma nas concentrações de amônia

A avaliação das doenças hepáticas nos felinos requer uma série de provas laboratoriais e as rotineiramente empregadas ainda são pouco elucidativas. Em face da escassez de dados e das controvérsias, foi realizada a determinação dos valores pré e pós-prandiais de ácidos biliares séricos (ABS) e de amônia plasmática (AP) e a avaliação da influência do período de estocagem. Os valores de ABS pré e pós-prandiais, por método enzimático-colorimétrico, de vinte gatos sadios, adultos, foram de 0,5 ± 0,1 e 3,6 ± 1,0 µmol/l, respectivamente. No que tange aos valores pré e pós-prandiais de AP obtidos 30 minutos após a colheita de sangue (157,0 ± 18,7 µg/dl ou 89,5 ± 10,7 µmol/l e 295,3 ± 28,2 µg/dl ou 165,2 ± 17,3 µmol/l), determinados pelo método eletrodo íon-específico, observou-se diferença estatisticamente significante, mas esta diferença deixou de existir ao se determinarem os níveis de amônia nas mesmas amostras estocadas (a -20ºC) por 24 e 48 horas. Esses resultados sugerem que a amostra de plasma não deve ser armazenada para posterior mensuração de amônia. Os valores observados para os ABS e AP poderão ser cotejados com aqueles obtidos em felinos com suspeita de afecção hepática, e assim auxiliar no diagnóstico e prognóstico.The evaluation of hepatic diseases in cats requires many laboratorial tests, and those routinely used are less conclusive. Considering the little availability of data and controversies related to the subject, we decided to study the preprandial and postprandial serum bile acids and plasma ammonia levels, and the effects of storage on these values. Preprandial (fasting) and postprandial serum bile acid (SBA) levels of 20 adult, healthy cats were determined using the enzymatic method. The measured values were 0.5 ± 0.1 and 3.6 ± 1.0 µmol/L, respectively. The preprandial and postprandial plasma ammonia (PA) levels, obtained 30 minutes after blood collection, 157.0 ± 18.7 µg/dL or 89.5 ± 10.7 µmol/L and 295.3 ± 28.2 µg/dL or 165.2 ± 17.3 µmol/L, were determined using the ion-specific electrode method, showing significant difference. However, this difference was not observed when the ammonia levels were measured in plasma samples kept under frozen storage (-20ºC) during 24 and 48 hours. These results suggest that the plasma samples should not be stored for further ammonia level determination. The SBA and PA levels measured for healthy cats in this study can be compared to those obtained from cats under suspicion of liver disease, helping to establish the diagnosis and prognosis

Stem cell differentiation for muscle regeneration

Physical activity has a positive role on muscle remodelling and vascularization, involv-ing stem cells differentiation processes. Indeed, the skeletal muscle homeostasis and repair are maintained by a subset of muscle stem/progenitor cells called Satellite Cells (SCs), while for heart repair and remodelling the cardiac potential of progenitor cells is otherwise expressed by different stem cell types: bone marrow hematopoietic stem cells (BMHSC), bone marrow mesenchymal stem cells (BMMSC), cardiac stem cells and embryonic stem cells. The ε isoform of the PKC family (PKCε) is a serine-threonine kinase that is expressed in muscle and in a variety of other tissues, regulating their homeostasis acting on cell death and differentiation. We focused on the role of PKCε in skeletal, cardiac and smooth muscle differentiation of adult stem cells. We found that inhibition of PKCε prevents myogenic differentiation of the myoblast cell line C2C12 and of primary SCs. In vivo PKCε inhibition resulted in impaired muscle regeneration, as well [1]. On the contrary, in cardiac and smooth muscle differentia-tion of stem cells we observed a negative role of PKCε both in vitro and in vivo [2,3]. In fact, it impaired cardiac markers expression like NKX2.5 and GATA4 but also vascular differ-entiation markers like SMA and PECAM. PKCε should therefore be considered as a finely tuned modulator of muscle cell differentiation

Ventilação controlada mecânica em cavalos com o emprego de vecurônio

O objetivo do presente estudo foi avaliar os efeitos metabólicos e respiratórios da administração de vecurônio em cavalos submetidos a ventilação controlada e compará-los àqueles que permaneceram em respiração espontânea. Foram empregados vinte animais hígidos alotados em dois grupos experimentais. Todos os animais foram pré-medicados com romifidina (100 mi g/kg IV) sendo a anestesia induzida com a associação de tiletamina-zolazepam (2 mg/kg IV) e a manutenção realizada com halotano. Os animais do grupo I permaneceram em respiração espontânea enquanto os animais do grupo II receberam vecurônio na dose de 0,1 mg/kg IV sendo submetidos a ventilação controlada mecânica. A administração do vecurônio não promoveu qualquer alteração significativa da freqüência ou ritmo cardíaco, pressão venosa central ou pressão arterial. No atinente aos animais que permaneceram em respiração espontânea, não houve qualquer diferença em relação a estes parâmetros quando comparados aos dos animais que permaneceram em respiração espontânea. Os animais que receberam o vecurônio apresentaram valores inferiores de PaCO2 e valores normais de pH em relação aos animais do grupo I. A duração de ação do vecurônio foi de 12,83 ± 1,72 minutos. Após o término da administração do halotano, os animais do grupo II retornaram à ventilação espontânea em 6,09 minutos demonstrando valores de PaCO2 da ordem de 50,78 mmHg. Não houve necessidade de reversão farmacológica do bloqueador e a qualidade da recuperação foi semelhante nos dois grupos. Frente aos resultados obtidos, pode-se concluir que o emprego de ventilação controlada mecânica e vecurônio em eqüinos é factível e isenta de efeitos adversos, sendo portanto indicada nesta espécie.The purpose of this study was to investigate the metabolic and cardiorespiratory effects of the administration of vecuronium under controlled mechanical ventilation versus spontaneous ventilation in horses. Twenty healthy horses scheduled to elective surgery were randomized and assigned in two groups. All animals were pre-medicated with romifidine (100 mu g/kg iv), anesthesia was induced with an association of tiletamine-zolazepam (2 mg/kg iv) and was maintained with halothane. Animals of group I remained in spontaneous ventilation without muscle relaxation while group II received vecuronium (0.1 mg/kg IV) and was submitted to controlled mechanical ventilation. Vecuronium administration did not cause any significant change of heart rate or rhythm, central venous pressure and arterial pressure. With the animals that remained under spontaneous ventilation, no differences were observed in these parameters. The animals that received vecuronium showed lower values of PaCO2 and normal values of pH in relation to the spontaneous ventilation group. Vecuronium duration was 12.83 ± 1.72 minutes. After halothane discontinuation the weaning time in group I I was 6.09 minutes with mean final PaCO2 values of 50.78 mmHg. There was no need of pharmacological reversion of vecuronium effect and recovery from anesthesia was similar in the two groups. In conclusion, the use of mechanical controlled ventilation and vecuronium in horses is easily performed, does not induce further cardiovascular depression and should be employed in equines undergoing major operations

Online teaching in physiotherapy education during COVID-19 pandemic in Italy: a retrospective case-control study on students\u2019 satisfaction and performance

Background: During COVID-19 pandemic, physiotherapy lecturers faced the challenge of rapidly shifting from face-to-face to online education. This retrospective case-control study aims to compare students' satisfaction and performances shown in an online course to a control group of students who underwent the same course delivered face-to-face in the previous five years. Methods: Between March and April 2020, a class (n = 46) of entry-level physiotherapy students (University of Verona - Italy), trained by an experienced physiotherapist, had 24-hours online lessons. Students exposed to the same course in the previous five academic years (n = 112), delivered with face-to-face conventional lessons, served as a historical control. The course was organized in 3 sequential phases: (1) PowerPoint presentations were uploaded to the University online platform, (2) asynchronous video recorded lectures were provided on the same platform, and (3) between online lectures, the lecturer and students could communicate through an email chat to promote understanding, dispel any doubts and collect requests for supplementary material (e.g., scientific articles, videos, webinars, podcasts). Outcomes were: (1) satisfaction as routinely measured by University with a national instrument and populated in a database; (2) performance as measured with an oral examination. Results: We compared satisfaction with the course, expressed on a 5-point Likert scale, resulting in no differences between online and face-to-face teaching (Kruskal-Wallis 2 = 0.24, df = 1, p = 0.62). We weighted up students' results by comparing their mean performances with the mean performances of the same course delivered face-to-face in the previous five years, founding a statistical significance in favour of online teaching (Wilcoxon rank sum test W = 1665, p < 0.001). Conclusions: Online teaching in entry-level Physiotherapy seems to be a feasible option to face COVID-19 pandemic, as satisfies students as well as face-to-face courses and leading to a similar performance. Entry-level Bachelors in Physiotherapy may consider moving to eLearning to facilitate access to higher education. Universities will have to train lecturers to help them develop appropriate pedagogical skills, and supply suitable support in terms of economic, organizational, and technological issues, aimed at guaranteeing a high level of education to their students. Trial registration: Retrospectively registered

Correlation between Protein Kinase Cε expression and thrombotic risk in Primary Myelofibrosis (PMFs)

Myelofibrosis (MF) - either primary (PMF) or arising from a previous PV or ET - is a Philadelphia-negative MPNs characterized by aberrant platelet production and consequent variable platelet count with altered hemostatic function (1). It has already been demonstrated that the risk of thrombotic events is one of the most common co-morbidities associated with PV and ET (2-5). However, risk of thrombotic events in PMF has not been investigated yet. We previously demonstrated that PKCepsilon (PKCε) is over-expressed in platelets from patients with acute myocardial infarction and accounts for their increased reactivity (6). Additionally, we recently showed that PKCε overexpression plays a crucial role in PMF MK impaired differentiation and that its levels correlated with the disease severity (expressed by the IPSS/DIPPS risk category) (7,8). On these bases, we analyzed PKCε expression in platelets from PMF patients, investigating a potential correlation with thrombotic risk and the aggressiveness of the disease. For this study, peripheral blood samples from 6 PMF patients and 3 healthy donors (HD) were collected in Na-citrate tubes. PKCε mRNA and protein levels were determined in platelets purified as described by Carubbi C, 2012. Finally, patients are stratified according to the history of cardio-vascular events and the IPSS/DIPSS risk category. PMF platelets showed significantly higher mRNA levels of PKCε as compared to HD. Protein analysis confirm PKCε over-expression in PMF platelets, almost reaching statistical significance. We then found that platelet from PMF patients who suffered from cardiovascular events display significantly higher levels of PKCε as compared to the one with a negative history. Finally, similarly to what observed in PMF magakaryocytes, we showed a positive correlation between PKCε platelets levels and IPSS/DIPSS risk category, with the lowest levels in low-risk patients and higher levels in high-risk patients. Collectively, our preliminary results indicate that PMF platelets show an aberrant expression of PKCε which correlates with the disease burden and a history of cardiovascular events. This suggests that the over-expression of PKCε may account for PMF platelet altered reactivity and function

PKC epsilon involvement in Th17 in vitro differentiation: implications in psoriasis pathogenesis

Psoriasis is a noncontagious, arytematous-squamose dermatitits affecting both sexes and all races. Although its exact etiology is largely unknown, it is now recognized as one of the most common immune-mediated disorders and several studies demonstrate an impairment of regulatory T-cells (Tregs) function and an up-regulation of IL-17 levels produced by T-helper 17 lymphocytes (Th17)(1,2). Protein kinase C epsilon (PKCε) is a serine/threonine kinase which plays a key role in the proliferation and differentiation of epidermal cells. We have previously demonstrated a role for PKCε in the pathogenesis of the autoimmune disease Hashimoto’s thyroiditis (3). PKCε is over-expressed in CD4+ T lymphocytes isolated from PBMC fraction in patients affected by this pathology and its forced down-modulation primed the TGF-mediated in vitro Treg polarization of human T CD4+ cells. Since it has been demonstrated that PKC-signalling is altered in psoriatic keratinocytes (4), we investigated the involvement of PKCε in Th17 in vitro differentiation and its potentially implication in immune response correlated to psoriasis. Using western blot and real time PCR, we have observed that PKCε protein levels and mRNA increase during Th17-lineage in vitro differentiation from naïve CD4+ T cells with a similar trend of Th17 markers of differentiation STAT3 and RoRyT. Moreover, PKCε overexpression significantly increases STAT3 and phosphorylated STAT3 levels, suggesting that PKCε boosts Th17 polarization. Thereafter, we sought to investigate PKCε expression in CD4+ lymphocytes obtained from peripheral blood of psoriatic patients and we observed that PKCε expression levels are significantly higher compared with healthy donors. Intriguingly, we observed a closely correlation of PKCε expression with PASI index, suggesting an involvement of the kinase with the severity of the disease. Collectively these data suggest that PKCε might be involved in Th17 differentiation, that it could be a key factor to regulate Th17 pathological expansion and therefore a potential psoriatic pharmacological target

Uso da solução hipertônica de cloreto de sódio a 7,5% no tratamento da hipotensão arterial decorrente da anestesia com halotano em eqüinos

Solução hipertônica de cloreto de sódio a 7,5% tem sido utilizada no tratamento do choque hipovolêmico. Sua ação é caracterizada pelo aumento do débito cardíaco, da pressão arterial sistêmica e do volume plasmático. O objetivo deste estudo foi avaliar a ação da solução hipertônica de cloreto de sódio a 7,5% (4 ml/kg) no tratamento da hipotensão arterial induzida pela anestesia geral com halotano em eqüinos. Foram utilizados 15 eqüinos adultos, de ambos os sexos e raças e pesos variados, encaminhados a processos cirúrgicos eletivos. A técnica anestésica constituiu-se de sedação com detomidina, indução anestésica com éter gliceril guaiacol, midazolam e quetamina e manutenção anestésica com halotano em oxigênio. Estabeleceu-se como valor mínimo para infusão de solução hipertônica de cloreto de sódio a 7,5%, pressão arterial média inferior a 60 mmHg. Foram avaliados os seguintes parâmetros: freqüência e ritmo cardíacos; pressão arterial sistólica, diastólica e média; freqüência respiratória; tempo de preenchimento capilar; gases sangüíneos, pH arterial, bicarbonato plasmático; saturação da oxi-hemoglobina; concentração plasmática de sódio e cloreto e hematócrito. Considerou-se o tempo zero como controle e os parâmetros foram aferidos aos 5, 15, 30 e 60 minutos após a infusão. Verificou-se aumento significativo dos valores da pressão arterial, a qual manteve-se elevada durante o período de observação. Os valores do hematócrito diminuíram significativamente demonstrando ter havido expansão volêmica. Houve discreta hipernatremia e hipercloremia. Os demais parâmetros permaneceram inalterados. Pode-se concluir que a solução hipertônica de cloreto de sódio a 7,5% mostrou-se efetiva, podendo ser utilizada com segurança nos processos hipotensivos desencadeados pelaanestesia geral com halotano.7.5% sodium chloride hypertonic solution is widely employed to improve hemodynamic parameters mainly treatment of hemorrhagic shock. Its action is characterized by an increment of cardiac output, systemic blood pressure and plasmatic volume. The aim of this study was the evaluation of 7.5% sodium chloride hypertonic solution in the treatment of hypotension due to general anesthesia with halothane in horses. Fifteen adult horses of different breeds and weights submitted to electivesurgeries were allocated in this trial.The animals received detomidine as premedication. Induction of anesthesia was performed with guaifenesin, midazolam and ketamine. Halothane in 100% oxygen was used as the maintenance agent. When the mean arterial blood pressure fell below 60 mmHg, 4 ml/kg of 7.5% sodium chloride hypertonic solution were administered intravenously. Heart rate and rhythm, systolic, mean and diastolic blood pressure, respiratory rate, capillary refill time, blood gases, arterial pH, bicarbonate, oxyhaemoglobin saturation, sodium and chloride plasma levels, as well as packed cell volume were evaluated after 5,15, 30 and 60 minutes hypertonic infusion. A significant increase of arterial blood pressure was verified after hypertonic administration. Values of blood pressure remained above control during all the recording period. Packed cell volume decreased significantly showing the improvement of plasmatic volume caused by this solution. Discrete hypernatremia and hyperchloremia were observed. The other parameters remained unaltered. We can conclude that 7.5% sodium chloride hypertonic solution is effective and may be employed in the treatment of hypotension during halothane anesthesia in equines