B139 Oct-4 transcription factors play an important role in maintaining the pluripotent state of embryonic stem cells and may prevent expression of genes activated during differentiation. Human Oct-4 isoform mRNAs encode proteins that have identical POU DNA binding domains and C-terminal domains, but differ in their amino-terminal domains. We report here the cloning and characterization of the human Oct-4B isoform. Human Oct-4B cDNA encodes a 265-amino acid protein with a predicted molecular mass of 30 kDa. ES cell-based complementation assays using ZHBTc4 ES cells showed that unlike human Oct-4A, Oct-4B cannot sustain ES cell self-renewal. In addition, Oct-4B does not bind to a probe carrying the Oct-4 consensus binding sequence and we demonstrate that two separate regions of its amino terminal domain are responsible for inhibiting DNA binding. We also demonstrate that Oct-4B is mainly localized to the cytoplasm. Overexpression of Oct-4B did not activate transcription from Oct-4-dependent promoters, whereas Oct-4A did as previously reported. Furthermore, transcriptional activation by human Oct-4A was not inhibited by co-expression of Oct-4B. Taken together, these data suggest that the DNA binding, transactivation, and abilities to confer self-renewal of the human Oct-4 isoforms differ.

The human oct-4 isoforms differ in their ability to confer self-renewal.

Vitis vinifera (grape) contains various compounds with acknowledged phytochemical and pharmacological properties. Among the different parts of the plant, pomace is of particular interest as a winemaking industry by-product. A characterization of the water extract from grape pomace from Montepulciano d’Abruzzo variety (Villamagna doc) was conducted, and the bioactive phenolic compounds were quantified through HPLC-DAD-MS analysis. HypoE22, a hypothalamic cell line, was challenged with an oxidative stimulus and exposed to different concentrations (1 µg/mL−1 mg/mL) of the pomace extract for 24, 48, and 72 h. In the same conditions, cells were exposed to the sole catechin, in a concentration range (5–500 ng/mL) consistent with the catechin level in the extract. Cell proliferation was investigated by MTT assay, dopamine release through HPLC-EC method, PGE2 amount by an ELISA kit, and expressions of neurotrophin brain-derived neurotrophic factor (BDNF) and of cyclooxygenase-2 (COX-2) by RT-PCR. The extract reverted the cytotoxicity exerted by the oxidative stimulus at all the experimental times in a dose-dependent manner, whereas the catechin was able to revert the oxidative stress-induced depletion of dopamine 48 h and 72 h after the stimulus. The extract and the catechin were also effective in preventing the downregulation of BDNF and the concomitant upregulation of COX-2 gene expression. In accordance, PGE2 release was augmented by the oxidative stress conditions and reverted by the administration of the water extract from grace pomace and catechin, which were equally effective. These results suggest that the neuroprotection induced by the extract could be ascribed, albeit partially, to its catechin content.

Phenolic Characterization and Neuroprotective Properties of Grape Pomace Extracts

Grape pomace (GP) represents a very reliable source of polyphenols because it could be found globally as a remnant of the wine industry. During the winemaking process, two types of GP are generated: red GP and white GP, according to the produced wine, red or white. Grape pomace represents a viable source of polyphenols, mainly flavanols, procyanidins anthocyanins, and resveratrol which possess antioxidant and anti-inflammatory activities. Multiple differences were observed between red and white GP in terms of their antioxidant and anti-inflammatory activity in both in vitro and in vivo studies. Although most studies are focused on the antioxidant and anti-inflammatory effect of red grape pomace, there are still many variables that need to be taken into consideration, as well as extensive study of the white GP. It was observed that in both in vitro and in vivo studies, the GP polyphenols have a direct antioxidant activity by acting as a free radical scavenger or donating a hydrogen atom. It also possesses an indirect antioxidant and anti-inflammatory activity by reducing mitochondrial reactive oxygen species (ROS) generation, malondialdehyde (MDA), tumor necrosis factor-alpha (TNF-α), interleukin-1-beta (IL-1β), interleukin-6 (IL-6), nuclear factor kappa-light-chain-enhancer of activated B cells (NF- κβ), and inhibitor of nuclear factor kappa-B kinase subunit beta (Iκκβ) levels or nitrate oxide-4 (NOX4) expression and by increasing the levels of antioxidants enzymes like superoxide dismutase (SOD), catalase (CAT) glutathione reductase (GRx) and glutathione peroxidase(GPx). Besides these activities, many beneficial effects in ischemic heart diseases were also observed, such as the maintenance of the ventricular function as close as possible to normal, and the prevention of infarcted area extension. In this context, this review intends to present the actual knowledge of grape pomace’s potential antioxidant and anti-inflammatory activity in ischemic heart disease, knowledge gathered from existing in vitro and in vivo studies focused on this.

https://www.mdpi.com/2227-9059/10/10/2337/pdf?version=1663675308

Antioxidant and Anti-Inflammatory Actions of Polyphenols from Red and White Grape Pomace in Ischemic Heart Diseases

Grape pomace is a source of anthocyanins, which can prevent cardiovascular diseases due to their antioxidant properties. Anthocyanin activity is associated with the ability to regulate oxidative stress through the transcription factor Nrf2. Thus, the present study aimed to evaluate if the anthocyanins found in Pinot noir pomace extract can affect the target genes related to the Nrf2 signalling pathway in endothelial cells. Our results highlight that the predominant anthocyanin in the Pinot noir pomace extract was malvidin-3-glucoside (3.7 ± 2.7 Eq. Malv-3-glu/kg). Molecular docking indicated that cyanidin-3-glucoside (−6.9 kcal/mol), malvidin-3-glucoside (−6.6 kcal/mol) and peonidin-3-glucoside (−6.6 kcal/mol) showed the highest affinities for the binding sites of the BTB domains in Keap1, suggesting that these components may modify the interaction of this protein with Nrf2. In addition, when HUVEC cells were exposed to different concentrations of Pinot noir pomace extract (100 µg/mL, 200 µg/mL, and 400 µg/mL), no changes in Nrf2 gene expression were observed. However, the gene expression of HO-1 and NQO1, which are in the signalling pathway of this transcription factor, increased according the concentrations of the extract (p = 0.0004 and p = 0.0084, respectively). In summary, our results show that anthocyanins play a very important role in Nrf2 activation and release, while at the same time not promoting its transcription. These preliminary results strongly suggest that the Pinot noir pomace extract can serve as a potent bioactive component source that protects cells against oxidative stress.

/pdf/anthocyanins-found-in-pinot-noir-waste-induce-target-genes-2gly5sc1.pdf

Anthocyanins Found in Pinot Noir Waste Induce Target Genes Related to the Nrf2 Signalling in Endothelial Cells

3-Bromofluoranthene-induced cardiotoxicity of zebrafish and apoptosis in the vascular endothelial cells via intrinsic and extrinsic caspase-dependent pathways.

TAP 1.0: A robust immunoinformatic tool for the prediction of tumor T-cell antigens based on AAindex properties.

Protective effect of Pinot noir pomace extract against the cytotoxicity induced by polycyclic aromatic hydrocarbons on endothelial cells.

Las celulas perivasculares tienen un origen comun en las celulas madre embrionarias y en los vasos sanguineos que proporcionan un nicho para la mantencion de su troncalidad La expresion de marcadores embrionarios y de celulas indiferenciadas, como tambien la gran variedad de fenotipos celulares generados desde los pericitos, podrian ser explicados por la capacidad de estas celulas de ser inducidas a un estado "stemness" cuando son tratadas con factores adecuados Nuestros resultados describen la expresion de celulas con OCT-4 citoplasmatico en una ubicacion anatomica perivascular donde de su nicho se encuentra en la region intima de la aorta en rata In vitro las celulas aisladas por el metodo de explante que promueve el aislamiento de celulas migratorias desde los tejidos muestran un fenotipo con un citoplasma alargado y que expresan aSMA, PDGFRa y b, siendo estos dos ultimos marcadores especificos de pericitos En estas celulas se presenta una tranlocacion a la variante nuclear de OCT-4 que ha sido descrito como el principal regulador de los procesos de autorrenovacion y pluripotencia La expresion de OCT-4 confirma y amplia aun mas las observaciones obtenidas en nuestras investigaciones anteriores y demuestra que celulas madre se encuentran en los vasos sanguineos en un microambiente que, probablemente, les permite que sobrevivan y permanezcan en reposo como un tipo de celula troncal quiescente

/pdf/la-translocacion-in-vitro-citoplasma-nucleo-del-factor-de-26a2a8b9gp.pdf

La Translocación in vitro Citoplasma/Núcleo del Factor de Transcripción Embrionario OCT-4 en Células Perivasculares Propone a la Aorta Como un Nicho Quiescente de Células Madres Adultas

HERRERA-BRAVO, J.; MONTIEL-EULEFI, E.; GLASER, T.; GARCES, M; LEAL, P. & HENNING, U. A. La Translocacion InVitro Citoplasma/Nucleo del Factor de Transcripcion Embrionario OCT-4 en Celulas Perivasculares Propone a la Arteria Aorta Como unNicho Quiescente de Celulas Madre Adultas. Int. J. Morphol., 31(4):1430-1438, 2013.RESUMEN: Las celulas perivasculares tienen un origen comun en las celulas madre embrionarias y en los vasos sanguineos queproporcionan un nicho para la mantencion de su troncalidad. La expresion de marcadores embrionarios y de celulas indiferenciadas,como tambien la gran variedad de fenotipos celulares generados desde los pericitos, podrian ser explicados por la capacidad de estascelulas de ser inducidas a un estado “stemness” cuando son tratadas con factores adecuados. Nuestros resultados describen la ex presionde celulas con OCT-4 citoplasmatico en una ubicacion anatomica perivascular donde de su nicho se encuentra en la region intima de laaorta en rata. In vitro las celulas aisladas por el metodo de explante que promueve el aislamiento de celulas migratorias desde los tejidosmuestran un fenotipo con un citoplasma alargado y que expresan aSMA, PDGFRa y b, siendo estos dos ultimos marcadores especifico sde pericitos. En estas celulas se presenta una tranlocacion a la variante nuclear de OCT-4 que ha sido descrito como el princip al reguladorde los procesos de autorrenovacion y pluripotencia. La expresion de OCT-4 confirma y amplia aun mas las observaciones obtenidas ennuestras investigaciones anteriores y demuestra que celulas madre se encuentran en los vasos sanguineos en un microambiente que,probablemente, les permite que sobrevivan y permanezcan en reposo como un tipo de celula troncal quiescente.PALABRAS CLAVE: Troncalidad; Pericitos; Translocacion; OCT-4; Aorta de rata.

/pdf/la-translocacion-in-vitro-citoplasma-nucleo-del-factor-de-1p4y5xfpuf.pdf

La Translocación in vitro Citoplasma/Núcleo del Factor de Transcripción Embrionario OCT-4 en Células Perivasculares Propone a la Aorta Como un Nicho Quiescente de Células Madres Adultas In vitro Translocation Cytoplasm/nucleus of Embryonic Transcription Factor OCT-4 in Perivascular Cells Suggests that Aorta as a Niche of Quiescent Adult Stem Cells

Jesús Herrera-Bravo

Papers

TAP 1.0: A robust immunoinformatic tool for the prediction of tumor T-cell antigens based on AAindex properties.

Protective effect of Pinot noir pomace extract against the cytotoxicity induced by polycyclic aromatic hydrocarbons on endothelial cells.

La Translocación in vitro Citoplasma/Núcleo del Factor de Transcripción Embrionario OCT-4 en Células Perivasculares Propone a la Aorta Como un Nicho Quiescente de Células Madres Adultas