Buffalo embryos produced by in vitro fertilization from oocytes matured during long-term transport

El búfalo (Bubalus bubalis) es una especie con excelente adaptación a sectores inundables. El mejoramiento genético a través de superovulación y transferencia embrionaria ha tenido escasos resultados debido a difcultades en la detección de celo, pobre respuesta ovárica y limitada recuperación de embriones post-lavaje. La técnica de fertilización in vitro de embriones (FIV) es una biotecnología de gran impacto en el progreso genético. El objetivo del presente trabajo fue estudiar los eventos tempranos de la FIV, analizando la tasa de maduración y desarrollo embrionario post-fertilización de ovocitos madurados in vitro (IVM) durante el transporte. Ovocitos bovinos y bubalinos fueron obtenidos por punción folicular de ovarios post-mortem e IVM durante el transporte por un período de 18 h. Se realizó la FIV con toros de fertilidad comprobada, con una concentración en microgotas de inseminación de 3-4 x 106 espermatozoides motiles/ml por un período de 6 horas. Los embriones fueron cultivados en medio oviductal sintético SOFaa en incubadora gaseada y ambiente humidificado a 38,5ºC durante 9 días. Se evaluaron las tasas de IVM, clivaje (día 2 post-fertilización) y blastocisto (días 7 a 9). Los resultados fueron analizados estadísticamente utilizando Fischer's Exact Test (p<0,05). No se observaron diferencias significativas en la tasa de maduración de ovocitos bubalinos de buena calidad respecto al control sin transporte (72 vs 88%), pero se registró una reducción significativa en la maduración de los ovocitos bubalinos de mala calidad (35%). Asimismo, se lograron producir los primeros embriones bubalinos luego de FIV, aunque las tasas de clivaje (34 vs 70 y 78%) y blastocisto (3 vs 27 y 31%) fueron significativamente menores en búfalos que en bovinos con y sin transporte, respectivamente. Los datos del presente trabajo constituirían el primer informe de FIV en búfalos y producción in vitro de embriones luego de IVM de ovocitos durante el transporte.The water buffalo (Bubalus bubalis) is a species with excellent adaptation to food-prone environments. Genetic improvement using the multiple ovulation and embryo transfer approach has been met with poor results in the buffalo, mainly due to difficulties in heat detection, erratic ovarian response to treatments and low embryo recovery post-flush. In vitro fertilization (IVF) is a powerful reproductive biotechnology that may provide a tool for genetic improvement in this species. The objective of this experiment was to study early embryonic events after IVF in the buffalo, analyzing in vitro maturation and IVF of oocytes matured during ground transportation. Bovine and bubaline oocytes were collected by follicular aspiration of post-mortem ovaries and in vitro matured for 18 h during ground transportation. In vitro fertilization was conducted, semen form bulls of proven fertility was processed and adjusted to a fnal concentration of 3-4 x 106 motile spermatozoa/ml in the insemination drops, oocytes were co-incubated for a period of 6 h. Embryos were then cultured in synthetic oviductal fluid (SOFaa) medium in an incubator and humidified atmosphere at 38.5ºC for 9 days. Oocyte maturation, cleavage and blastocyst rates were evaluated on days 0, 2 and 7 to 9, respectively and results were statistically analyzed using Fischer´s Exact Test (p<0.05). No significant difference was observed in the maturation rate of bubaline oocytes of good quality vs the non-transported control (72 vs 88%); however, the maturation rate of bubaline oocytes of bad quality was significantly lower (35%) than the rest of the groups. Data of present experiment are the first report of buffalo embryos produced by IVF from oocytes matured during transportation, although the cleavage (34 vs 70 and 78%) and blastocyst (3 vs 27 and 31%) rates were significantly lower for the buffalo than for the transported and non-transported domestic cattle, respectively.Fil: Konrad, José Luis. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Scian, R.. Pont. Universidad Catolica Arg."sta.maria de Los Bs.as.". Facultad de Cs.agrarias; ArgentinaFil: Garrido, María José. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; ArgentinaFil: Taminelli, G.. Pont. Universidad Catolica Arg."sta.maria de Los Bs.as.". Facultad de Cs.agrarias; ArgentinaFil: Sansinena, Marina Julia. Pont. Universidad Catolica Arg."sta.maria de Los Bs.as.". Facultad de Cs.agrarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Campos de precipitación de la provincia del Chubut (periodo 1931-1960)

Durante varios años el Centro Nacional Patagónico ha recogido información sobre precipitación de diversas fuentes y períodos, con particular énfasis en la proveniente de pobladores rurales, disponiendo actualmente de datos de más de cien localidades del Chubut, provenientes en general de diferentes períodos y presentando a veces series incompletas. Trabajos anteriores de los autores mostraron que la precipitación ha variado durante el presente siglo en forma diferente en distintas regiones de la Provincia; aun así, métodos elaborados en el Centro Nacional Patagónico permiten incluir series de períodos cortos (menos de 30 años)en el análisis de los campos de precipitación en el período elegido 1931-1960. Se utilizan resultados de otros trabajos de los autores para completar series defectuosas, y se trazan mapas estacionales y anuales de precipitación en base a la información citada y a las características del relieve. En lugares con información escasa o nula se utilizan datos de fuera del período 1931-1960, como referenciaThe Centro Nacional Patagónico has gathered precipitation data of over a hundred locations of Chubut. Many of them are coming from countrymen. The data covers different periods and most of them are incomplete. In a former paper, the authors have shown that precipitation has undergone significant changes during this century in a different way for different regions. A method developed in the Centro Nacional Patagónico, allows the assimilation of short series. Annual and stational maps of mean precipitation of Chubut for the normal period of thirty years (1931-1960) are shown.Asociación Argentina de Geofísicos y Geodesta

Producción de embriones de búfalo por fertilización in vitro luego de la maduración de los ovocitos durante el transporte prolongado

El búfalo (Bubalus bubalis) es una especie con excelente adaptación a sectores inundables. El mejoramiento genético a través de superovulación y transferencia embrionaria ha tenido escasos resultados debido a dificultades en la detección de celo, pobre respuesta ovárica y limitada recuperación de embriones post-lavaje. La técnica de fertilización in vitro de embriones (FIV) es una biotecnología de gran impacto en el progreso genético. El objetivo del presente trabajo fue estudiar los eventos tempranos de la FIV, analizando la tasa de maduración y desarrollo embrionario post-fertilización de ovocitos madurados in vitro (IVM) durante el transporte. Ovocitos bovinos y bubalinos fueron obtenidos por punción folicular de ovarios post-mortem e IVM durante el transporte por un período de 18 h. Se realizó la FIV con toros de fertilidad comprobada, con una concentración en microgotas de inseminación de 3-4 x 106 espermatozoides motiles/ml por un período de 6 horas. Los embriones fueron cultivados en medio oviductal sintético SOFaa en incubadora gaseada y ambiente humidificado a 38,5ºC durante 9 días. Se evaluaron las tasas de IVM, clivaje (día 2 post-fertilización) y blastocisto (días 7 a 9). Los resultados fueron analizados estadísticamente utilizando Fischer’s Exact Test (

Expression signatures of TP53 mutations in serous ovarian cancers

<p>Abstract</p> <p>Background</p> <p>Mutations in the <it>TP53 </it>gene are extremely common and occur very early in the progression of serous ovarian cancers. Gene expression patterns that relate to mutational status may provide insight into the etiology and biology of the disease.</p> <p>Methods</p> <p>The <it>TP53 </it>coding region was sequenced in 89 frozen serous ovarian cancers, 40 early stage (I/II) and 49 advanced stage (III/IV). Affymetrix U133A expression data was used to define gene expression patterns by mutation, type of mutation, and cancer stage.</p> <p>Results</p> <p>Missense or chain terminating (null) mutations in <it>TP53 </it>were found in 59/89 (66%) ovarian cancers. Early stage cancers had a significantly higher rate of null mutations than late stage disease (38% vs. 8%, p < 0.03). In advanced stage cases, mutations were more prevalent in short term survivors than long term survivors (81% vs. 30%, p = 0.0004). Gene expression patterns had a robust ability to predict <it>TP53 </it>status within training data. By using early versus late stage disease for out of sample predictions, the signature derived from early stage cancers could accurately (86%) predict mutation status of late stage cancers.</p> <p>Conclusions</p> <p>This represents the first attempt to define a genomic signature of <it>TP53 </it>mutation in ovarian cancer. Patterns of gene expression characteristic of <it>TP53 </it>mutation could be discerned and included several genes that are known p53 targets or have been described in the context of expression signatures of <it>TP53 </it>mutation in breast cancer.</p

N-Octanoyl-Dopamine inhibits cytokine production in activated T-cells and diminishes MHC-class-II expression as well as adhesion molecules in IFN gamma-stimulated endothelial cells

IFN gamma enhances allograft immunogenicity and facilitates T-cell mediated rejection. This may cause interstitial fibrosis and tubular atrophy (IFTA), contributing to chronic allograft loss. We assessed if inhibition of T-cell activation by N-octanoyl dopamine (NOD) impairs adherence of activated T-cells to endothelial cells and the ability of activated T-cells to produce IFN gamma. We also assessed if NOD affects IFN gamma mediated gene expression in endothelial cells. The presence of NOD during T-cell activation significantly blunted their adhesion to unstimulated and cytokine stimulated HUVEC. Supernatants of these T-cells displayed significantly lower concentrations of TNF alpha and IFN gamma and were less capable to facilitate T-cell adhesion. In the presence of NOD VLA-4 (CD49d/CD29) and LFA-1 (CD11a/CD18) expression on T-cells was reduced. NOD treatment of IFN gamma stimulated HUVEC reduced the expression of MHC class II transactivator (CIITA), of MHC class II and its associated invariant chain CD74. Since IFTA is associated with T-cell mediated rejection and IFN gamma to a large extent regulates immunogenicity of allografts, our current data suggest a potential clinical use of NOD in the treatment of transplant recipients. Further in vivo studies are warranted to confirm these in vitro findings and to assess the benefit of NOD on IFTA in clinically relevant models

Small Molecule Inhibited Parathyroid Hormone Mediated cAMP Response by N–Terminal Peptide Binding

Ligand binding to certain classes of G protein coupled receptors (GPCRs) stimulates the rapid synthesis of cAMP through G protein. Human parathyroid hormone (PTH), a member of class B GPCRs, binds to its receptor via its N–terminal domain, thereby activating the pathway to this secondary messenger inside cells. Presently, GPCRs are the target of many pharmaceuticals however, these drugs target only a small fraction of structurally known GPCRs (about 10%). Coordination complexes are gaining interest due to their wide applications in the medicinal field. In the present studies we explored the potential of a coordination complex of Zn(II) and anthracenyl–terpyridine as a modulator of the parathyroid hormone response. Preferential interactions at the N–terminal domain of the peptide hormone were manifested by suppressed cAMP generation inside the cells. These observations contribute a regulatory component to the current GPCR–cAMP paradigm, where not the receptor itself, but the activating hormone is a target. To our knowledge, this is the first report about a coordination complex modulating GPCR activity at the level of deactivating its agonist. Developing such molecules might help in the control of pathogenic PTH function such as hyperparathyroidism, where control of excess hormonal activity is essentially required

Status of the ENUBET Project

The ENUBET Collaboration is designing the first “monitored neutrino beam”: a beam with an unprecedented control of the flux, energy and flavor of neutrinos at source. In particular, ENUBET monitors the νe production mostly by the detection of large angle positrons from three body semileptonic decays of kaons: K+ → e+π0νe. In this paper, we present the status of the Project and the 2018-2019 advances on proton extraction, transfer line, particle identification in the decay tunnel and beam performance

Mutant p53 drives multinucleation and invasion through a process that is suppressed by ANKRD11

Mutations of p53 in cancer can result in a gain of function associated with tumour progression and metastasis. We show that inducible expression of several p53 ‘hotspot’ mutants promote a range of centrosome abnormalities, including centrosome amplification, increased centrosome size and loss of cohesion, which lead to mitotic defects and multinucleation. These mutant p53-expressing cells also show a change in morphology and enhanced invasive capabilities. Consequently, we sought for a means to specifically target the function of mutant p53 in cancer cells. This study has identified ANKRD11 as a key regulator of the oncogenic potential of mutant p53. Loss of ANKRD11 expression with p53 mutation defines breast cancer patients with poor prognosis. ANKRD11 alleviates the mitotic defects driven by mutant p53 and suppresses mutant p53-mediated mesenchymal-like transformation and invasion. Mechanistically, we show that ANKRD11 restores a native conformation to the mutant p53 protein and causes dissociation of the mutant p53–p63 complex. This represents the first evidence of an endogenous protein with the capacity to suppress the oncogenic properties of mutant p53.JE Noll, J Jeffery, F Al-Ejeh, R Kumar, KK Khanna, DF Callen and PM Neilse