ROGDI is a novel gene which has unknown function. According to GenBank, the gene is located on chromosome 16p13.3 and the size of coding region is 864 bp which encodes 287 amino acids. It was a novel gene isolated from primary human renal epithelial cells in NEDO human cDNA sequencing project (AK026039). The definition of its reference sequence (RefSeq NM_024589) described as Homo sapiens rogdi homolog (Drosophila), ROGDI. By bioinformatic analysis, the gene was predicted as a hydrophilic protein with leucine zipper domain and located in cytoplasm. A partial cDNA of this gene was cloned in our laboratory. For further study of the biological function of the gene, the coding region of this gene was cloned into pGEX-6p and pET-28a vector and expressed in E. coli BL21 (DE3). The fusion protein was partially purified for preparation of polyclonal antibody. Northern blot analysis revealed that the gene was not expressed in all tissues. From the results of RT-PCR and western blot analysis, it can be concluded that the products, both mRNA and protein, of this gene were found in many cancer cell lines, but protein level expression of the gene was much less in normal cell lines. By immunocytochemistry analysis and subcellular localization analysis of GFP-tagged ROGDI, the gene was expressed both in the nucleus and cytoplasm, but expressed more in the nucleus than cytoplasm. In addition, ROGDI was up-regulated in early stages of liver fibrosis of TAA-treated mouse livers. This novel gene may play roles in tumorigenesis and liver fibrosis.

OBJETIVO: Determinar los predictores no invasivos de la severidad de la fibrosis hepática en pacientes con infección crónica por hepatitis C del Servicio de Gastroenterología del Hospital IV Alberto Sabogal (EsSALUD), durante el período diciembre 2000 – diciembre 2002. MATERIAL Y MÉTODOS: Se incluyeron a 30 pacientes con serología positiva para HCV que fueron sometidos a Laparoscopia más biopsia hepática. Estudio observacional, descriptivo, retrospectivo y de corte transversal. Se usó la base de datos del programa SPSS versión 10.0, y se evaluó la asociación mediante el test de Chi cuadrado (_i2), con una significancia estadística del 5%. RESULTADOS: La edad promedio fue de 56.27 ± 12.55 años, Se demostró asociación entre fibrosis severa y radio TGO/TGP mayor de 1 y esplenomegalia(p=0.018 y 0.001, respectivamente) en forma estadísticamente significativa,mientras que la plaquetopenia menor de 150,000 no mostró asociación con fibrosis hepática severa en el grupo de pacientes estudiados (p=0.136). CONCLUSIONES: El radio TGO/TGP mayor de 1 y la esplenomegalia pueden ser considerados como predictores de fibrosis avanzada en pacientes con infección crónica por hepatitis c. En estos pacientes quizás no sea necesaria una biopsia hepática. / OBJETIVE: To Determine the non-invasive predictors from Hepatic Fibrosis severity in patients with Hepatitis C chronic infection at Gastroenterology Service from Alberto Sabogal Hospital (EsSALUD), since december 2000 – december 2002. MATERIAL AND METHODS: 30 patients were included with positive HCV serology who had Laparoscopy and hepatic biopsy. This is an observational, descriptive, retrospective and transversal study. It was used SPSS 10.0 database program, and was evaluated its association with chi-square test (_i2), with 5% statistical significance. RESULTS: The age average was 56.27 ± 12.55 years old. A TGO/TGP ratio of major of 1 and splenomegaly correlated significantly with advance stage of fibrosis (p=0.018 and 0.001 respectively) but there wasn’t correlation between platelet count major of 150,000 and severe fibrosis (p=0.136). CONCLUSIONS: A TGO/TGP ratio of major of 1 and splenomegalia can predict advance stage of fibrosis in patients with chronic hepatitis C infection .In these patients, a liver biopsy may not be necessary.

Liver fibrosis is an important reversible stage in progress of most chronic liver diseases (CLDs). The excess hepatic wound healing response against chronic liver injury results in extracellular matrix proteins accumulation and fibrosis. Oxidative stress, liver inflammation and/or hepatic steatosis contribute to this process. Until now, little is known how dietary lipids can influence liver’s pathophysiology. The effects of lipids on CLD progression may depend on their amount and the quality of fatty acids as well as the degrees of saturation. The investigation of liver fibrosis will help to understand the pathogenesis of CLDs and develop potential nutritional therapeutic approaches. The specific aim of this study was to investigate the effects of different high fats consumption in liver fibrosis by feeding the normal and carbon tetrachloride (CCl4)-treated animals with the diets enriched with following oils: corn oil rich in ω-6 polyunsaturated fatty acids (PUFAs), extra virgin olive oil (EVOO) high in ω-9 monounsaturated fatty acids (MUFAs), and lard enriched with saturated fatty acids (SFAs) for 4 weeks. The differentially expressed liver proteins in this process were identified by two-dimensional gel electrophoresis based proteomics to explore the molecular mechanisms. The proteomic analysis revealed characteristic differences between (i) normal and fibrotic livers (Chapter 3), and between the fibrotic livers treated with (ii) low fat versus high fat (20% w/w corn oil, Chapter 4) and among the high fats, between the diet enriched with corn oil versus (iii) EVOO (Chapter 5) and lard (Chapter 6). Among the identified proteins, collagen synthesis related protein prolyl 4-hydroxylase, oxidative stress related protein alpha-1-antitrypsin, free radical scavenger Cu/Zn superoxide dismutase and Calcium homeostasis regulator calreticulin and regucalcin were found to involve in CCl4-induced liver fibrosis. The results show that corn oil enhanced the hepatic steatosis but had no significant effects on fibrogenesis; the expression of several stress proteins like heat shock protein 75 kDa, and lipid metabolism related protein enoyl-CoA hydratase domain-containing protein 3 were found increased in high corn oil consumption animals with CCl4-treatment. Histological evaluations showed that olive oil could attenuate, and lard oil aggravate the liver damage induced by CCl4. Compared to corn oil, high EVOO diet rich in MUFAs decreased the lipid peroxidation and collagen accumulation in liver. Several protein related to antioxidant effects, including peroxiredoxin-1, thiosulfate sulfurtransferase and thioredoxin domain-containing protein 12 were found have higher expression level in high EVOO intake animals. In contrast, lard rich in SFAs intake leaded to macrovesicular steatosis and advanced fibrosis, and decreased the expression of antioxidant related glutathione S-transferases. Interestingly, S-adenosylmethionine synthesis related enzyme methionine adenosyltransferase was found up-regulated in lard intake animals, suggests the modification of DNA methylation was implicated in lard fed animals, while the demethylation on the promoter of profibrogenic gene was found, confirmed the lard consumption has the epigenetic modification effects in liver injury. Together, these findings give further insight into the pathobiology of CLDs. The data also helped to address the issue that different degrees of saturation of dietary lipids may affect liver fibrosis with different mechanistic actions. / published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

The effectiveness of liposomes incorporating bismuth-ethanedithiol and loaded with tobramycin (LipoBiEDT-TOB) at sub-inhibitory concentrations to inhibit the production of quorum sensing signaling molecules and virulence factors induced by P. aeruginosa was evaluated in vitro. In addition, we evaluated the efficacy and safety of free and encapsulated tobramycin in liposomal formulations administered intratracheally to rats chronically infected with P. aeruginosa. LipoBiEDT-TOB significantly reduced the production of quorum sensing signaling molecules and virulence factor secretion compared to free tobramycin. The LipoBiEDT-TOB formulation significantly reduced the bacterial count in lungs, modulated the IL-8 level in blood and minimized the nephrotoxicity that is associated with aminoglycoside treatment. These results support the hypothesis that aerosolization of liposomal aminoglycosides may enhance the management of chronic lung infections caused by resistant P. aeruginosa in patients with cystic fibrosis.

No description available.

No description available.

No description available.

No description available.

Cardiac fibrosis is the excess deposition of myocardial extracellular matrix components, which increases tissue stiffness and heterogeneity, causing impaired diastolic/systolic function and arrhythmias, and eventually leading to heart failure and death. There are no available treatments for cardiac fibrosis. Myofibroblasts mediate fibrosis, and are characterized by hypersynthesis of collagens, decreased migration, and increased α-smooth muscle actin, which is incorporated into stress fibers, imparting contractility. Scleraxis is a transcriptional regulator of collagen-rich tissues, increased in response to the same stimuli that drive the myofibroblast phenotype, such as cyclic stretch. We show that Scleraxis mediates the conversion of cardiac fibroblasts to myofibroblasts, by increasing myofibroblast marker expression and contraction, and decreasing migration. Additionally, a proximal 1500 bp human SCLERAXIS promoter is activated by stretch and is responsive to transforming growth factor-β1. Thus, Scleraxis is a specific mechanoresponsive regulator of the myofibroblast, representing a novel target for the treatment of cardiac fibrosis.

Liver transplantation was first performed in 1963 (1) as an experimental treatment for end stage liver disease. Three patients were transplanted, all of whom died within 3 weeks. Since then it has become an established therapy resulting in improved quality of life (2), with 675 transplants from cadaveric donors taking place in the UK in 2001 and 706 in 2002 (3). This level of activity compares with 10 years ago when 502 liver transplants were performed in 1992. Figures released for survival up to the year 2000 show that early (1 year) survival has improved to 88% for patients transplanted from 1998 – 1999, with 3 year survival for the period 1996 – 1997 being 73% and 5 year survival for the period 1994 – 1995 being 64% (3). This improvement is probably due to a combination of factors such as improved surgical and anaesthetic technique, changes in medical management after transplantation, the improved recognition of other harmful factors like hypertension, choice of immune suppression and better prediction of patients in whom liver transplantation is not likely to be appropriate such as those with cholangiocarcinoma or multiple large hepatocellular carcinomas. [Taken from Introduction]