Sorafenib prevents liver fibrosis in a non-alcoholic steatohepatitis (NASH) rodent model

Liver fibrosis occurring as an outcome of non-alcoholic steatohepatitis (NASH) can precede the development of cirrhosis. We investigated the effects of sorafenib in preventing liver fibrosis in a rodent model of NASH. Adult Sprague-Dawley rats were fed a choline-deficient high-fat diet and exposed to diethylnitrosamine for 6 weeks. The NASH group (n=10) received vehicle and the sorafenib group (n=10) received 2.5 mg·kg-1·day-1 by gavage. A control group (n=4) received only standard diet and vehicle. Following treatment, animals were sacrificed and liver tissue was collected for histologic examination, mRNA isolation, and analysis of mitochondrial function. Genes related to fibrosis (MMP9, TIMP1, TIMP2), oxidative stress (HSP60, HSP90, GST), and mitochondrial biogenesis (PGC1α) were evaluated by real-time quantitative polymerase chain reaction (RT-qPCR). Liver mitochondrial oxidation activity was measured by a polarographic method, and cytokines by enzyme-linked immunosorbent assay (ELISA). Sorafenib treatment restored mitochondrial function and reduced collagen deposition by nearly 63% compared to the NASH group. Sorafenib upregulated PGC1α and MMP9 and reduced TIMP1 and TIMP2 mRNA and IL-6 and IL-10 protein expression. There were no differences in HSP60, HSP90 and GST expression. Sorafenib modulated PGC1α expression, improved mitochondrial respiration and prevented collagen deposition. It may, therefore, be useful in the treatment of liver fibrosis in NASH.

Avaliação de extratos de quebra- pedra (Phyllanthus sp) frente à patógenos causadores de infecções no trato urinário / Evaluation of Phyllanthus sp extracts in face of Pathogen Causers of Urinary Treat Infections

RESUMO O trato urinário normalmente é estéril, no entanto, pode ser contaminado por agentes etiológicos provenientes da microbiota intestinal, dentre os mais comuns pode-se destacar a Escherichia coli. Os microrganismos estão se tornando cada vez mais resistentes a múltiplos antimicrobianos. É notória, portanto, a necessidade de encontrar novas substâncias com propriedades antimicrobianas. Portanto, foram avaliados diferentes extratos de Phyllanthus sp, frente a microrganismos causadores de infecções no trato urinário e comparadas as técnicas de hole-plate e disco difusão. Para ambas as técnicas avaliadas, o extrato de 72 horas mostrou melhor atividade antimicrobiana, na técnica de disco difusão, a bactéria mais sensível foi o Staphylococcus saprophyticcus, que apresentou CMI (Concentração Mínima Inibitória) de 15,84 mg/mL. Com a utilização da técnica de hole-plate, a bactéria mais sensível observada foi Staphylococcus aureus, com valor de CMI igual ao reportado anteriormente. Foi observado que os extratos alcoólicos obtiveram maior eficiência em relação às infusões, que a técnica de hole-plate revelou ser mais eficiente que a técnica de disco difusão e que os cocos Gram positivos foram mais susceptíveis quando comparadas aos bacilos Gram negativos e fungos.

ABSTRACT Commonly, the urinary treat is sterile, but it can also be contaminated by etiological agents from the intestinal treat, of which the Escherichiacoli is the most common one. These microorganisms are increasingly becoming more resistant to multiple antibiotics. It became necessary to find new substances with antimicrobial properties, so the purpose of this paper is to evaluate different Phyllanthus sp extracts- in face of microorganisms which cause the urinary treat infections- and compare it to the hole-plate and disk diffusion techniques. The 72 hours extraction showed better antimicrobial activity in both methods. Using disk diffusion, the most sensitive bacterium was the Staphylococcus saprophyticcus, with the MIC of 15,84 mg/mL. While using the technique of hole-plate, the most sensitive bacterium was the Staphylococcus aureus, with the same MIC of the previous cited bacterium. This study showed that the alcoholic extracts were more efficient than the infusions. It can also be observed that the hole-plate technique seems to be more efficient than the disk diffusion one, and the Gram positive cocci bacteria were more sensitive than the Gram negative bacilli and fungi.

MTP -493G/T gene polymorphism is associated with steatosis in hepatitis C-infected patients

The reduction of hepatic microsomal transfer protein (MTP) activity results in fatty liver, worsening hepatic steatosis and fibrosis in chronic hepatitis C (CHC). The G allele of the MTP gene promoter, -493G/T, has been associated with lower transcriptional activity than the T allele. We investigated this association with metabolic and histological variables in patients with CHC. A total of 174 untreated patients with CHC were genotyped for MTP -493G/T by direct sequencing using PCR. All patients were negative for markers of Wilson’s disease, hemochromatosis and autoimmune diseases and had current and past daily alcohol intake lower than 100 g/week. The sample distribution was in Hardy-Weinberg equilibrium. Among subjects with genotype 1, 56.8 percent of the patients with fibrosis grade 3+4 presented at least one G allele versus 34.3 percent of the patients with fibrosis grade 1+2 (OR = 1.8; 95 percentCI = 1.3-2.3). Logistic regression analysis with steatosis as the dependent variable identified genotypes GG+GT as independent protective factors against steatosis (OR = 0.4, 95 percentCI = 0.2-0.8; P = 0.01). The results suggest that the presence of the G allele of MTP -493G/T associated with lower hepatic MTP expression protects against steatosis in our CHC patients.

Does hepatocellular carcinoma in non-alcoholic steatohepatitis exist in cirrhotic and non-cirrhotic patients?

Non-alcoholic steatohepatitis (NASH) has been associated with hepatocellular carcinoma (HCC) often arising in histologically advanced disease when steatohepatitis is not active (cryptogenic cirrhosis). Our objective was to characterize patients with HCC and active, histologically defined steatohepatitis. Among 394 patients with HCC detected by ultrasound imaging over 8 years and staged by the Barcelona Clinic Liver Cancer (BCLC) criteria, we identified 7 cases (1.7 percent) with HCC occurring in the setting of active biopsy-proven NASH. All were negative for other liver diseases such as hepatitis C, hepatitis B, autoimmune hepatitis, Wilson disease, and hemochromatosis. The patients (4 males and 3 females, age 63 ± 13 years) were either overweight (4) or obese (3); 57 percent were diabetic and 28.5 percent had dyslipidemia. Cirrhosis was present in 6 of 7 patients, but 1 patient had well-differentiated HCC in the setting of NASH without cirrhosis (fibrosis stage 1) based on repeated liver biopsies, the absence of portal hypertension by clinical and radiographic evaluations and by direct surgical inspection. Among the cirrhotic patients, 71.4 percent were clinically staged as Child A and 14.2 percent as Child B. Tumor size ranged from 1.0 to 5.2 cm and 5 of 7 patients were classified as early stage; 46 percent of all nodules were hyper-echoic and 57 percent were <3 cm. HCC was well differentiated in 1/6 and moderately differentiated in 5/6. Alpha-fetoprotein was <100 ng/mL in all patients. HCC in patients with active steatohepatitis is often multifocal, may precede clinically advanced disease and occurs without diagnostic levels of alpha-fetoprotein. Importantly, HCC may occur in NASH in the absence of cirrhosis. More aggressive screening of NASH patients may be warranted.

Liver mitochondrial dysfunction and oxidative stress in the pathogenesis of experimental nonalcoholic fatty liver disease

Oxidative stress and hepatic mitochondria play a role in the pathogenesis of nonalcoholic fatty liver disease. The aim of the present study was to evaluate the role of hepatic mitochondrial dysfunction and oxidative stress in the pathogenesis of the disease. Fatty liver was induced in Wistar rats with a choline-deficient diet (CD; N = 7) or a high-fat diet enriched with PUFAs-omega-3 (H; N = 7) for 4 weeks. The control group (N = 7) was fed a standard diet. Liver mitochondrial oxidation and phosphorylation were measured polarographically and oxidative stress was estimated on the basis of malondialdehyde and glutathione concentrations. Moderate macrovacuolar liver steatosis was observed in the CD group and mild liver steatosis was observed in the periportal area in the H group. There was an increase in the oxygen consumption rate by liver mitochondria in respiratory state 4 (S4) and a decrease in respiratory control rate (RCR) in the CD group (S4: 32.70 ± 3.35; RCR: 2.55 ± 0.15 ng atoms of O2 min-1 mg protein-1) when compared to the H and control groups (S4: 23.09 ± 1.53, 17.04 ± 2.03, RCR: 3.15 ± 0.15, 3.68 ± 0.15 ng atoms of O2 min-1 mg protein-1, respectively), P < 0.05. Hepatic lipoperoxide concentrations were significantly increased and the concentration of reduced glutathione was significantly reduced in the CD group. A choline-deficient diet causes moderate steatosis with disruption of liver mitochondrial function and increased oxidative stress. These data suggest that lipid peroxidation products can impair the flow of electrons along the respiratory chain, causing overreduction of respiratory chain components and enhanced mitochondrial reactive oxygen species. These findings are important in the pathogenesis of nonalcoholic fatty liver disease.

Avaliação da seleçäo feminina de basquete / Evaluation of Brazilian female basketball team

Foram avaliadas 13 jogadoras de basquete convocadas para disputa do Jogos Olímpicos em Sidney - 2000. As atletas foram submetidas à mensuração do peso e altura, a um teste ergoespirométrico em esteira e a um teste de impulsão vertical. Todos os testes foram realizados no Cemafe (Centro de Medicina da Atividade Física e do Esporte) da Unifesp/EPM, em junho de 2000. Os resultados encontrados estão descritos a seguir: idade média de 24,4ñ4,6 anos, peso corporal 70,8ñ8,6kg, estatura 182ñ7,6 cm, e índice de massa corpórea (IMC) 21,4ñ1,8. A avaliação ergoespirométrica revelou valores para o consumo máximo de oxigênio (VO2 máx) de 49,9ñ5,4 ml/kg/min; o consumo de oxigênio no limiar anaeróbico (VO2 LA) de 38,7ñ4,3ml/kg/min; frequência cardíaca máxima (FC máx) 194,4ñ9,6bpm; frequência cardíaca no limiar anaeróbico (FC LA) 179,7ñ5,7bpm; velocidade no limiar anaeróbico (VEL LA) 119,9ñ1,4km/h; gasto calórico no limiar anaeróbico (GASTO CAL) 815,6ñ71,8kcal/h. No teste de impulsão vertical as atletas foram orientadas a saltar com a maior potência e o maior número de vezes possível em 30 segundos, detectando-se os índices por equipamento sonar (Vertisonic©). Foram encontrados para a potência máxima ou pico de potência 9,9ñ2,2W e um trabalho muscular total durante os 30 segundos de 602,1ñ104,8J. A avaliação médica e fisiológica é fundamental para atletas de alto nível alcançarem sucesso em competições de nível internacional, sendo estes parâmetros essenciais ao planejamento e periodização do treinamento