Erk1/2 signaling mediates the HGF-induced protection against ethanol and acetaldehyde-induced toxicity in the pancreatic RINm5F cell line.

Alcohol-induced pancreas damage remains as one of the main risk factors for pancreatitis development. This disorder is poorly understood, particularly the effect of acetaldehyde, the primary alcohol metabolite, in the endocrine pancreas. Hepatocyte growth factor (HGF) is a protective protein in many tissues, displaying antioxidant, antiapoptotic, and proliferative responses. In the present work, we were focused on characterizing the response induced by HGF and its protective mechanism in the RINm5F pancreatic cell line treated with ethanol and acetaldehyde. RINm5F cells were treated with ethanol or acetaldehyde for 12 h in the presence or not of HGF (50 ng/ml). Cells under HGF treatment decreased the content of reactive oxygen species and lipid peroxidation induced by both toxics, improving cell viability. This effect was correlated to an improvement in insulin expression impaired by ethanol and acetaldehyde. Using a specific inhibitor of Erk1/2 abrogated the effects elicited by the growth factor. In conclusion, the work provides mechanistic evidence of the HGF-induced-protective response to the alcohol-induced damage in the main cellular component of the endocrine pancreas.

Bik subcellular localization in response to oxidative stress induced by chemotherapy, in Two different breast cancer cell lines and a Non-tumorigenic epithelial cell line.

Cancer chemotherapy remains one of the preferred therapeutic modalities against malignancies despite its damaging side effects. An expected outcome while utilizing chemotherapy is apoptosis induction. This is mainly regulated by a group of proteins known as the Bcl-2 family, usually found within the endoplasmic reticulum or the mitochondria. Recently, these proteins have been located in other sites and non-canonic functions have been unraveled. Bik is a pro-apoptotic protein, which becomes deregulated in cancer, and as apoptosis is associated with oxidative stress generation, our objective was to determine the subcellular localization of Bik either after a direct oxidative insult due to H2 O2 , or indirectly by cisplatin, an antineoplastic agent. Experiments were performed in two human transformed mammary gland cell lines MDA-MB-231 and MCF-7, and one non-tumorigenic epithelial cell line MCF-10A. Our results showed that in MCF-7, Bik is localized within the cytosol and that after oxidative stress treatment it translocates into the nucleus. However, in MDA-MB-231, Bik localizes in the nucleus and translocates to the cytosol. In MCF10A Bik did not change its cellular site after either treatment. Interestingly, MCF10A were more resistant to cisplatin than transformed cell lines. This is the first report showing that Bik is located in different cellular compartments depending on the cancer stage, and it has the ability to change its subcellular localization in response to oxidative stress. This is associated with increased sensitivity when exposed to toxic agents, thus rendering novel opportunities to study new therapeutic targets allowing the development of more active and less harmful agents.

Normal development of the heart: a review of new findings.

Development and formation of the heart, the central organ of the circulatory system in vertebrates, starts early during embryonic development (second week), reaching maturity during the first few postnatal months. Cardiogenesis is a highly complex process that requires the active and orderly participation of different cardiac and non-cardiac cell populations. Thus, this process is sensitive to errors that may trigger a variety of heart-development defects, called congenital heart defects, which have a worldwide incidence of 8-10/1000 live births. A good understanding of normal cardiogenesis is required for better diagnosis and treatment of congenital heart diseases. This article reviews normal cardiogenesis by comparing information from classic studies with more recent findings. Information from descriptive anatomical studies of histological sections and selective in vivo marking of chicken embryos were emphasized. In addition, the discovery of heart fields has fueled the investigation of cardiogenic events that were believed to be understood and has contributed to proposals for new models of heart development.

El corazón, órgano central del aparato circulatorio de los vertebrados, comienza a formarse muy temprano en el desarrollo embrionario (segunda semana de gestación) y alcanza su forma madura durante los primeros meses posteriores al nacimiento. La cardiogénesis se caracteriza por ser un proceso altamente complejo, dependiente de la participación activa y ordenada de diferentes poblaciones celulares cardiacas y no cardiacas. Lo anterior hace que este proceso sea sensible a errores que pueden desencadenar una variedad de defectos del desarrollo cardiaco, llamados cardiopatías congénitas, con una incidencia mundial de 8 a 10/1000 nacidos vivos. Para mejorar el diagnóstico y el tratamiento de las cardiopatías congénitas es necesario comprender adecuadamente los eventos implicados en la cardiogénesis normal. En este artículo se revisa el desarrollo cardiaco normal, contrastando la información de los estudios clásicos con la de hallazgos recientes. Se hace hincapié en la información obtenida de los estudios de anatomía descriptiva de cortes histológicos y marcaje selectivo in vivo en embriones de pollo. Adicionalmente, el descubrimiento de los campos cardiogénicos ha estimulado la investigación de eventos cardiogénicos que se creían comprendidos, contribuyendo con propuestas de nuevos modelos del desarrollo del corazón.

HGF induces protective effects in α-naphthylisothiocyanate-induced intrahepatic cholestasis by counteracting oxidative stress.

Cholestasis is a clinical syndrome common to a large number of hepatopathies, in which either bile production or its transit through the biliary tract is impaired due to functional or obstructive causes; the consequent intracellular retention of toxic biliary constituents generates parenchyma damage, largely via oxidative stress-mediated mechanisms. Hepatocyte growth factor (HGF) and its receptor c-Met represent one of the main systems for liver repair damage and defense against hepatotoxic factors, leading to an antioxidant and repair response. In this study, we evaluated the capability of HGF to counteract the damage caused by the model cholestatic agent, α-naphthyl isothiocyanate (ANIT). HGF had clear anti-cholestatic effects, as apparent from the improvement in both bile flow and liver function test. Histology examination revealed a significant reduction of injured areas. HGF also preserved the tight-junctional structure. These anticholestatic effects were associated with the induction of basolateral efflux ABC transporters, which facilitates extrusion of toxic biliary compounds and its further alternative depuration via urine. The biliary epithelium seems to have been also preserved, as suggested by normalization in serum GGT levels, CFTR expression and cholangyocyte primary cilium structure our results clearly show for the first time that HGF protects the liver from a cholestatic injury.

Normal development of the heart: a review of new findings / Desarrollo normal del corazón: revisión de nuevos hallazgos

Abstract Development and formation of the heart, the central organ of the circulatory system in vertebrates, starts early during embryonic development (second week), reaching maturity during the first few postnatal months. Cardiogenesis is a highly complex process that requires the active and orderly participation of different cardiac and non-cardiac cell populations. Thus, this process is sensitive to errors that may trigger a variety of heart-development defects, called congenital heart defects, which have a worldwide incidence of 8-10/1000 live births. A good understanding of normal cardiogenesis is required for better diagnosis and treatment of congenital heart diseases. This article reviews normal cardiogenesis by comparing information from classic studies with more recent findings. Information from descriptive anatomical studies of histological sections and selective in vivo marking of chicken embryos were emphasized. In addition, the discovery of heart fields has fueled the investigation of cardiogenic events that were believed to be understood and has contributed to proposals for new models of heart development.

Resumen El corazón, órgano central del aparato circulatorio de los vertebrados, comienza a formarse muy temprano en el desarrollo embrionario (segunda semana de gestación) y alcanza su forma madura durante los primeros meses posteriores al nacimiento. La cardiogénesis se caracteriza por ser un proceso altamente complejo, dependiente de la participación activa y ordenada de diferentes poblaciones celulares cardiacas y no cardiacas. Lo anterior hace que este proceso sea sensible a errores que pueden desencadenar una variedad de defectos del desarrollo cardiaco, llamados cardiopatías congénitas, con una incidencia mundial de 8 a 10/1000 nacidos vivos. Para mejorar el diagnóstico y el tratamiento de las cardiopatías congénitas es necesario comprender adecuadamente los eventos implicados en la cardiogénesis normal. En este artículo se revisa el desarrollo cardiaco normal, contrastando la información de los estudios clásicos con la de hallazgos recientes. Se hace hincapié en la información obtenida de los estudios de anatomía descriptiva de cortes histológicos y marcaje selectivo in vivo en embriones de pollo. Adicionalmente, el descubrimiento de los campos cardiogénicos ha estimulado la investigación de eventos cardiogénicos que se creían comprendidos, contribuyendo con propuestas de nuevos modelos del desarrollo del corazón.

The proximal segment of the embryonic outflow (conus) does not participate in aortic vestibule development.

OBJECTIVE: There is no consensus on the embryonic components or morphogenetic processes involved in mature ventricular outflow tract development. Our goal was to use in vivo labelling to investigate the prospective fate of the myocardium of each conal wall. The conal and atrioventricular cushion mesenchyme changes during transformation into mature structures and their role in apoptosis were also investigated. METHODS: Plastic labels were placed at the cephalic and caudal conal limits of chicken embryo hearts (stage 22HH) and traced up to stage 36HH. Histological analyses, scanning electron microscopy and apoptotic detection using Lysotracker-Red were performed. The conal longitudinal length and medial displacement were registered. Muscle myosin was identified by immunofluorescence. RESULTS: Labels positioned in the myocardium of each conal wall moved to the right ventricle (RV), shifting from the arterial subvalvular myocardial zone to the apex. No labels were found in the aortic vestibule. At stage 22HH, the conus was a tubular structure composed of myocardium and endocardium with scarce mesenchyme. The dorso-left conal myocardial wall gradually lost continuity and the free ends separated, while the myocardium was distributed to the RV free wall (24HH-28HH). At stage 22HH, conal crests were not observed, but they were apparent at the dorsal zone of the conus at stage 26HH; towards stage 30HH, they fused to form the supraventricular crest, and the pulmonary infundibulum was evident. The ventro-superior cushion of the AV canal was reorganized into the fibrous and muscular structures lined the aortic vestibule. CONCLUSIONS: The posterior conus is an erroneous concept. The conal myocardium is reorganized in the free wall of the RV. Internally, the conal lumen is transformed into the pulmonary infundibulum. The aortic vestibule is formed from the ventro-superior cushion of the AV canal. Thus, the ventricular outflow tracts have different embryonic origins.

Molecular Cloning of a cDNA Encoding for Taenia solium TATA-Box Binding Protein 1 (TsTBP1) and Study of Its Interactions with the TATA-Box of Actin 5 and Typical 2-Cys Peroxiredoxin Genes.

TATA-box binding protein (TBP) is an essential regulatory transcription factor for the TATA-box and TATA-box-less gene promoters. We report the cloning and characterization of a full-length cDNA that encodes a Taenia solium TATA-box binding protein 1 (TsTBP1). Deduced amino acid composition from its nucleotide sequence revealed that encodes a protein of 238 residues with a predicted molecular weight of 26.7 kDa, and a theoretical pI of 10.6. The NH2-terminal domain shows no conservation when compared with to pig and human TBP1s. However, it shows high conservation in size and amino acid identity with taeniids TBP1s. In contrast, the TsTBP1 COOH-terminal domain is highly conserved among organisms, and contains the amino acids involved in interactions with the TATA-box, as well as with TFIIA and TFIIB. In silico TsTBP1 modeling reveals that the COOH-terminal domain forms the classical saddle structure of the TBP family, with one α-helix at the end, not present in pig and human. Native TsTBP1 was detected in T. solium cysticerci´s nuclear extract by western blot using rabbit antibodies generated against two synthetic peptides located in the NH2 and COOH-terminal domains of TsTBP1. These antibodies, through immunofluorescence technique, identified the TBP1 in the nucleus of cells that form the bladder wall of cysticerci of Taenia crassiceps, an organism close related to T. solium. Electrophoretic mobility shift assays using nuclear extracts from T. solium cysticerci and antibodies against the NH2-terminal domain of TsTBP1 showed the interaction of native TsTBP1 with the TATA-box present in T. solium actin 5 (pAT5) and 2-Cys peroxiredoxin (Ts2-CysPrx) gene promoters; in contrast, when antibodies against the anti-COOH-terminal domain of TsTBP1 were used, they inhibited the binding of TsTBP1 to the TATA-box of the pAT5 promoter gene.

Primary cultured astrocytes from old rats are capable to activate the Nrf2 response against MPP+ toxicity after tBHQ pretreatment.

Astrocytes are key players for brain physiology, protecting neurons by releasing antioxidant enzymes; however, they are also susceptible to damage by neurotoxins. Nuclear factor erythroid-derived 2-like 2 (Nrf2) is a central regulator of the antioxidant response, and therefore, pharmacologic inducers are often used to activate this transcription factor to induce cellular protection. To date, it still remains unknown if cells from aged animals are capable of developing this response. Therefore, the purpose of this work was to determine if cortical astrocytes derived from old rats are able to respond to tertbuthyl-hydroquinene (tBHQ) pretreatment and stimulate the Nrf2-antioxidant response pathway to induce an antioxidant strategy against MPP+ toxicity, one of the most used molecules to model Parkinson's disease. Our results show that, although astrocytes from adult and old rats were more susceptible to MPP+ toxicity than astrocytes from newborn rats, when pretreated with tertbuthyl-hydroquinene, they were able to transactivate Nrf2, increasing antioxidant enzymes and developing cellular protection. These results are discussed in terms of the doses used to create protective responses.