Further evidence on the role of heparan sulfate as protamine acceptor during the decondensation of human spermatozoa

Background: Human spermatozoa decondense in vitro upon exposure to heparin and glutathione. Glutathione is also the disulfide bond reducer in vivo, and heparan sulfate, a functional analogue of heparin, has been proposed as the protamine acceptor. The aim of this study was to evaluate the decondensing ability of chemically modified heparins and different glycosaminoglycans (GAGs) on isolated sperm nuclei in vitro, and to analyse the possible role of different GAGs as protamine acceptors. Methods: Capacitated spermatozoa and isolated sperm nuclei from normospermic semen samples were decondensed in the presence of heparin (or its equivalent) and glutathione. After fixation with glutaraldehyde, the percentage of decondensed spermatozoa and nuclei was determined under phase-contrast. Proteins were extracted from sperm nuclei previously incubated in the presence of gluhathione and different GAGs by incubation with urea-β-meracptoethanol-NaCl, and analysed by acid polyacrylamide gel electrophoresis. Results: The ability of desulfated heparins and other GAGs to decondense isolated nuclei mirrored exactly the decondensation of capacitated spermatozoa, the only difference being the level of maximum decondensation achieved. Heparan sulfate and heparin, but not other GAGs, were able to release protamines from sperm chromatin. Conclusions: Heparan sulfate could be functioning as protamine acceptor in vivo during human sperm nuclear decondensation. © The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.Fil:Romanato, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Regueira, E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

MDCK cells express serotonin-regulable 11β-hydroxysteroid dehydrogenase type 2

Prior to this work, we found that adrenal as well as extra-adrenal factors activate the response of renal l 11β-hydroxysteroid dehydrogenase 2 to stressful situations. These results -showing ways through which the organism hinders the pathological occupation of mineralocorticoid receptors by glucocorticoids leading to sodium retention and hypertension- prompted the present study on the nature of the above-mentioned extra-adrenal factors. Serotonin was chosen because of its properties as a widely distributed neurohormone, known to interact with glucocorticoids at many sites, also exhibiting increased levels and effects under stressful situations. We studied serotonin effects on 11β-hydroxysteroid dehydrogenase 2 activity in a cell line derived from distal nephron polarized-epithelium, employing 3H-corticosterone as substrate. The end-product, 3H-11- dehydrocorticosterone was separated from the substrate by HPLC and quantified. Serotonin stimulated 11β-hydroxysteroid dehydrogenase 2 activity only at 2nM and 25pM, the magnitude of the response depending also on substrate concentration. The stimulation was blocked by the specific inhibitors methiothepin and ketanserin. We postulate that the organism partially prevents renal mineralocorticoid receptor occupancy by glucocorticoids, circulating at enhanced levels under stressful situations, through serotonin-mediated catabolic regulation of the 11β-hydroxysteroid dehydrogenase 2 activity. Given many, mostly positive, interactions between both hormones, this might eventually pave the way to studies on a new regulatory axis.Fil:Zallocchi, M.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Matkovic', L.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Fábricas diagenéticas asociadas al paleokarsts del techo de la unidad intermedia del Mioceno de la Cuenca de Madrid.

Los materiales carbonáticos y evaporíticos del techo de la Unidad Intermedia del Mioceno de la Cuenca de Madrid han actuado como soporte de fenómenos de karstificación temprana, cuyo desarrollo ha supuesto un cambio significativo en la evolución del relleno de la cuenca. Dentro de estos materiales se han reconocido carbonatos con fábricas diagenéticas complejas resultado de procesos de diagénesis meteórica superficial. Los procesos predominantes en la evolución diagenética del encajante de la paleokarstificación del techo de la Unidad Intermedia son la dedolomitización y la calcitización/pseudomorfización de yesos. Factores como la composición original del sedimento, la influencia de microorganismos y la hidroquímica de los fluidos diagenéticos han determinado la gran variabilidad de las fábricas diagenéticas reconocidas. [ABSTRACT] Carbonates and evaporites from the top of the Miocene Intermediate Unit of the Madrid Basin have set as host-rock for early karstification phenomena. The development of these karstic processes imply an important change in the depositional evolution of the basin. In these materials, complex diagenetic fabrics formed as a result of shallow meteoric diagenetic processes have been recognized, being dedolomitization and calcitization of gypsums the main diagenetic processes that affected the top of the Miocene Intermediate Unit. The high variability of diagenetic fabrics is thought to be mainly influenced by the original composition of sediments, the activity of microorganisms and the chemistry of diagenetic waters

Leptin promotes cell proliferation and survival of trophoblastic cells

Leptin, the 16-kDa protein product of the obese gene, was originally considered as an adipocyte-derived signaling molecule for the central control of metabolism. However, leptin has been suggested to be involved in other functions during pregnancy, particularly in placenta. In the present work, we studied a possible effect of leptin on trophoblastic cell proliferation, survival, and apoptosis. Recombinant human leptin added to JEG-3 and BeWo choriocarcinoma cell lines showed a stimulatory effect on cell proliferation up to 3 and 2.4 times, respectively, measured by 3H-thymidine incorporation and cell counting. These effects were time and dose dependent. Maximal effect was achieved at 250 ng leptin/ml for JEG-3 cells and 50 ng leptin/ml for BeWo cells. Moreover, by inhibiting endogenous leptin expression with 2 μM of an antisense oligonucleotide (AS), cell proliferation was diminished. We analyzed cell population distribution during the different stages of cell cycle by fluorescence-activated cell sorting, and we found that leptin treatment displaced the cells towards a G2/M phase. We also found that leptin upregulated cyclin D1 expression, one of the key cell cycle-signaling proteins. Since proliferation and death processes are intimately related, the effect of leptin on cell apoptosis was investigated. Treatment with 2 μM leptin AS increased the number of apoptotic cells 60 times, as assessed by annexin V-fluorescein isothiocyanate/propidium iodide staining, and the caspase-3 activity was increased more than 2 fold. This effect was prevented by the addition of 100 ng leptin/ml. In conclusion, we provide evidence that suggests that leptin is a trophic and mitogenic factor for trophoblastic cells by virtue of its inhibiting apoptosis and promoting proliferation. © 2007 by the Society for the Study of Reproduction, Inc.Fil:Kotler, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Varone, C.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Influence of the electric energy non-regulated market in the intensive aquaculture plants associated to cooling effluents

En este trabajo se analiza el efecto que la liberalización del mercado eléctrico tiene sobre la variación de los regímenes de temperatura del agua en plantas de acuicultura intensiva que aprovechan los efluentes de refrigeración de centrales generadoras de electricidad. Para ello se han utilizado datos de una instalación dedicada al engorde de anguilas europeas, la cual toma el agua caliente del efluente de refrigeración de la Central Térmica de Puente Nuevo (Córdoba). Los resultados indican que la liberalización del mercado del sector eléctrico tiene una influencia significativa sobre la forma y cantidad de energía generada por la Central Térmica, y por consiguiente sobre el régimen termal del efluente de refrigeración. Los niveles de temperatura en el interior de la instalación son dependientes asimismo de la temperatura del agua en el efluente de refrigeración, estimándose la disminución de los índices de crecimiento debidos a este factor en un 5%.In this paper, the effect of the electric energy non-regulated market in the water thermal regimes variation of intensive fishfarms that use the heated water for cooling of power plants is analysed. This way, data of aneel intensive rearing system was used. In this fishfarm the heated water is drawn from the cooling effluent of the Puente Nuevo power plant (Córdoba). The results show that the non-regulated market has a significant effect on the form and amount of generated energy and the thermal regime of the cooling effluent. The temperature levels in the fishfarm depend of the water temperature of cooling effluent, being estimated the decrease of the growth index in 5%

Análisis sobre el origen de los materiales arqueológicos del Museo Histórico Minero D. Felipe de Borbón y Grecia, de la Escuela Técnica Superior de Ingenieros de Minas

En 1992 iniciamos el estudio de las piezas arqueológicas del Museo Histórico Minero D. Felipe de Borbón y Grecia, de la Escuela Técnica Superior de Ingenieros de Minas de Madrid, comprobando la calidad de las muestras presentes. Existían dos colecciones de Arqueología Minera y Prehistórica, ubicadas principalmente en las vitrinas del patio y tambié'f1 en las del torreón norte (1). Asimismo constatamos la presencia, en este último lugar, de un importante conjunto de piezas de la mina del Milagro, en anís, algunas en relación con las colecciones antes mencionadas

17Beta-estradiol enhances leptin expression in human placental cells through genomic and nongenomic actions

The process of embryo implantation and trophoblast invasion is considered the most limiting factor in the establishment of pregnancy. Leptin was originally described as an adipocyte-derived signaling molecule for the central control of metabolism. However, it has been suggested that leptin is involved in other functions during pregnancy, particularly in the placenta, where it was found to be expressed. In the present work, we have found a stimulatory effect of 17beta-estradiol (E2) on endogenous leptin expression, as analyzed by Western blot, in both the BeWo choriocarcinoma cell line and normal placental explants. This effect was time and dose dependent. Maximal effect was achieved at 10 nM in BeWo cells and 1 nM in placental explants. The E 2 effects involved the estrogen receptor, as the antagonist ICI 182 780 inhibited E2-induced leptin expression. Moreover, E2 treatment enhanced leptin promoter activity up to 4-fold, as evaluated by transient transfection with a plasmid construction containing the leptin promoter region and the reporter gene luciferase. This effect was dose dependent. Deletion analysis demonstrated that a minimal promoter region between - 1951 and -1847 bp is both necessary and sufficient to achieve E2 effects. Estradiol action involved estrogen receptor 1, previously known as estrogen receptor alpha, as cotransfection with a vector encoding estrogen receptor 1 potentiated the effects of E2 on leptin expression. Moreover, E2 action probably involves membrane receptors too, as treatment with an estradiol-bovine serum albumin complex partially enhanced leptin expression. The effects of E2 could be blocked by pharmacologic inhibition of MAPK and the phosphoinositide-3-kinase (PI3K) pathways with 50 μM PD98059 and 0.1 μM Wortmannin, respectively. Moreover, cotransfection of dominant negative mutants of MAP2K or MAPK blocked E 2 induction of leptin promoter. On the other hand, E2 treatment promoted MAPK1/MAPK3 and AKT phosphorylation in placental cells. In conclusion, we provide evidence suggesting that E2 induces leptin expression in trophoblastic cells, probably through genomic and nongenomic actions via crosstalk between estrogen receptor 1 and MAPK and PI3K signal transduction pathways. © 2010 by the Society for the Study of Reproduction, Inc.Fil:Gambino, Y.P. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Maymó, J.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Calvo, J.C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Varone, C.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Hipodoncia: utilidad de hallazgos dentales en la determinación de patologías y parentesco en poblaciones antiguas. Estudio de dos casos en la necrópolis oriental de Carthago Spartaria

The use of forensic techniques when studying partial or badly-conserved human remains is very important when we want to determine age, sex, and race, as well as parental relation between two or more individuals. Of these techniques, some of the simplest but most reliable are those related to the dental studies of the retrieved remains. Finding dental anomalies may give useful information about diseases suffered by the individual (congenital or infecto-contagious) and possible family relation between several individuals. This study describes several cases of dental agenesia (absence of teeth caused by lack of formation of these teeth) found in populations living during the fi fth to seventh centuries (a.D.), retrieved in the oriental necropolis of Carthago Spartaria, in the neighborhood of the present University (Cartagena, Murcia).La utilización de técnicas forenses a la hora de estudiar restos humanos parciales o en mal estado de conservación son muy importantes cuando queremos determinar la edad, sexo y raza, así como el parentesco entre dos o más individuos. De estas técnicas, algunas de las más sencillas pero de resultados bastante fi ables son aquellas relacionadas con el estudio dental de los restos encontrados. El hallazgo de anomalías dentarias puede facilitar información acerca de enfermedades padecidas por el individuo (tanto infectocontagiosas como congénitas) y de posibles relaciones familiares entre varios individuos. El presente estudio describe varios casos de agenesias dentales (ausencia dental debida a la falta de formación de algún diente) encontradas en una población de los siglos V-VII d.C., recuperados en la necrópolis oriental de Carthago Spartaria, en la zona del actual Barrio Universitario. (Cartagena, Murcia

A functional variant in the stearoyl-CoA desaturase (SCD) gene promoter affects gene expression in ovine muscle

The nutritional quality of lambs may be improved with increased stearoyl-CoA desaturase (SCD) gene expression, which increases the desaturation of stearic acid to oleic acid. The aim of this study was to evaluate the effect of the rs412429481 (FJ513370: g.31C > A) SNP located at the SCD gene on the functionality of the gene in lambs reared under different production systems. The effect of the rs412429481 SNP on gene expression in Rasa Aragonesa male lambs slaughtered at 22–24 kg was studied in two experiments. In Experiment 1 (n = 44), the semitendinosus muscle of lambs grazing alfalfa (ALF) or fed concentrates indoors (IND) was analysed; in Experiment 2 (n = 48), the semitendinosus and longissimus thoracis muscles of lambs that received supplementation with dl-a-tocopheryl acetate for different finishing periods were used. In Experiment 1, the effect of the rs412429481 SNP on the expression of the SCD gene in the semitendinosus muscle depended on the feeding group (P < 0.001), as it had no effect in ALF lambs, but CA lambs had greater SCD expression than CC lambs under the IND conditions. Moreover, ALF lambs showed lower levels of SCD gene expression than IND lambs (P < 0.05). In Experiment 2, gene expression was affected by the rs412429481 SNP in both muscles. Animals carrying the C- allele showed a lower expression rate than animals carrying the A- allele. These different expression levels were not associated with changes in the DNA methylation pattern or by the binding of specific nuclear proteins. Finally, we confirmed these results by luciferase assays, demonstrating that the SCD promoter containing the A variant had a 23.9% higher activity than the promoter containing the C variant