Effect of gonadotrophins, oestradiol and insulin on cumulus expansion of Nili Ravi buffalo oocytes

The objective of the present study was to investigate the cumulus expansions of Nili Ravi buffalo oocytes during cultured in TCM -199 supplemented with 2 micro g/ml oestradiol [E[2]], 0.05 IU/ml recombinant human follicle stimulating hormone [rhFSH], 2IU/ml human chorionic gonadotrophin [hCG], and 0.12 IU/ml insulin [I]. The cumulus oocytes complexes [COCs] were collected from 2-8mm follicles from local abattoir ovaries. Supplementation of medium with single hormones showed significant [P<0.0001] increase in mean diameter of COCs with rhFSH except E[2], hCG and insulin after 24 hours compared to the increase in the mean diameter of COCs matured in TCM-199 without any hormonal supplementation. With rhFSH even at 8th hour, significant increase [P<0.001] in cumulus expansion was observed. In combination of hormones the significant [P<0.0001] cumulus expansion was achieved in E[2]+rhFSH treatment group. The non significant [P>0.05] cumulus expansion was observed in treatment groups viz. E[2]+hCG, E[2]+Insulin, rhFSH+hCG, rhFSH+Insulin, hCG+Insulin, E[2]+rhFSH+hCG and E[2]+rhFSH+hCG+Insulin after 24 hours. In conclusion, supplementation of rhFSH alone and in combination with E2in TCM-199 has highly significant effect on cumulus expansion

Ultrastructural changes of erythrocytes in whole blood after exposure to prospective in silico-designed anticancer agents: a qualitative case study

BACKGROUND: Novel, in silico-designed anticancer compounds were synthesized in our laboratory namely, 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10),15-tetraen-17-ol (ESE-15-ol) and 2-ethyl-3-O-sulphamoyl-estra-1,3,5(10)16-tetraene (ESE-16). These compounds were designed to have improved bioavailability when compared to their source compound, 2-methoxyestradiol. This theoretically would be due to their increased binding affinity to carbonic anhydrase II, present in erythrocytes. Since the novel compounds under investigation are proposed to be transported within erythrocytes bound to carbonic anhydrase II, the morphological effect which they may exert on whole blood and erythrocytes is of great significance. A secondary outcome included revision of previously reported procedures for the handling of the whole blood sample. The purpose of this study was twofold. Firstly, the ultrastructural morphology of a healthy female's erythrocytes was examined via scanning electron microscopy (SEM) after exposure to the newly in silico-designed compounds. Morphology of erythrocytes following exposure to ESE-15-ol and ESE-16 for 3 minutes and 24 hours at 22°C were described with the use of SEM. The haemolytic activity of the compounds after 24 hours exposure were also determined with the ex vivo haemolysis assay. Secondly, storage conditions of the whole blood sample were investigated by determining morphological changes after a 24 hour storage period at 22°C and 37°C. RESULTS: No significant morphological changes were observed in the erythrocyte morphology after exposure to the novel anticancer compounds. Storage of the whole blood samples at 37°C for 24 hours resulted in visible morphological stress in the erythrocytes. Erythrocytes incubated at 22°C for 24 hours showed no structural deformity or distress. CONCLUSIONS: From this research the optimal temperature for ex vivo exposure of whole blood samples to ESE-15-ol and ESE-16 for 24 hours was determined to be 22°C. Data from this study revealed the potential of these compounds to be applied to ex vivo study techniques, since no damage occurred to erythrocytes ultrastructure under these conditions. As no structural changes were observed in erythrocytes exposed to ESE-15-ol and ESE-16, further ex vivo experiments will be conducted into the potential effects of these compounds on whole blood. Optimal incubation conditions up to 24 hours for whole blood were established as a secondary outcome.

Comparación de la acción antioxidante de estriol y estradiol in vitro sobre lipoproteínas de baja densidad en mujeres postmenopáusicas / Comparison of antioxidant actions of estriol and estradiol on low density lipoproteins in post menopausal women

Background: Estradiol (E2) has a potent antioxidant effect on low density lipoproteins (LDL) in vitro and in vivo, which could be important in explaining the cardioprotective effect of hormone replacement therapy (HRT) in post menopausal women. Estriol (E3), on the other hand, is a weak estrogen with low metabolic effects on different tissues, and at present no cardioprotective effect has been attributed to this steroid. Aim: To study the antioxidant effect of E3 on LDL and to compare it with the potent antioxidant action exhibited by E2. Subjects and methods: After LDL was isolated by ultra centrifugation from plasma of 12 healthy untreated post menopausal women, it was divided into aliquots containing 0.5 mg of LDL protein. Estriol and E2 in doses of 0, 1, 5, 15 and 50 µM were incubated with different aliquots of LDL. CuSO4 15 µM was added to each aliquot to induce an oxidative stress. The aliquots were then incubated during 4 hours at 37 ­C. Malonaldehyde (MDA) was measured as a marker of LDL oxidation, and expressed as nM/mg protein. Results (mean ñ SD): Estriol induced a dose-dependent decrease in MDA concentration (baseline 62.8 ñ 21.7; 1 µM: 61.5 ñ 23.0; 5 µM: 52.9 ñ 20,3; 15 µM 43.5 ñ 20.1 and 50 µM: 31.0 ñ 17.6 nM/mg protein; F= 92.4; p< 0.0001), reaching a mean decrease of 50.7 percent at the highest dose tested. Estradiol has a similar dose-dependent decrease in MDA concentration (F= 60.2; p< 0.0001), revealing a more potent effect than E3 (p< 0.05), with a mean decrease of 67.4 percent at the highest dose tested. Conclusions: Our results demonstrate that estriol shows an important antioxidant action of LDL in vitro, although its effect is less potent than estradiol. These results raise the possibility that estriol could have a cardioprotective effect in post menopausal women, possibility that has not been yet demonstrated

Modificaciones de los patrones mamográficos con diversos esquemas de hormonoterapia de reemplazo en mujeres postmenopáusicas / Modification of breast radiographic patterns with several hormonal therapy schemes in postmenopausal women

La mamografía constituye el método de mayor sensibilidad en el diagnóstico precoz de cáncer de mama y en especial en las mujeres mayores, con mamas tipo "Radiolúcidas". Es de indicación absoluta en mujeres candidatas a recibir HTR y para su seguimiento. La relación entre la HTR y el riesgo de desarrollar un cáncer de mama es un tema controversial aún no resuelto. Sin embargo estudios recientes señalan que la HTR puede inducir un aumento de la densidad mamográfica (DxMx). El aumento de la DxMx ha sido identificado como uno de los factores asociados a fallas en el Screening de cáncer de mama. El objetivo de nuestro estudio, realizado en 2 centros mamográficos privados, fue evaluar las modificaciones de la DxMx en 210 pacientes que recibían distintos esquemas de HTR durante 1 año de observación. Se dividieron en 7 grupos terapéuticos de 30 pacientes cada uno y se compararon contra un grupo control de similares características. Para cuantificar las modificaciones de la DxMx se usaron los patrones de Wolfe. Considerando los 7 grupos de tratamiento en forma global se obtuvo un aumento de la DxMx en un 32,3 por ciento. El grupo Tibolona y Estriol presentó el menor porcentaje de cambios en la DxMx y fue similar al grupo control sin HTR. El mayor porcentaje de aumento en la DxMx se observó en los grupos que recibieron valerianato de estradiol (E2) en forma exclusiva o en esquema secuencial con MPA (66,7 por ciento) y 56,7 por ciento respectivamente). El grupo que recibió estrógenos conjugados en forma exclusiva mostró un aumento de la DxMx de un 43 por ciento y el grupo con estrógenos conjugados más MPA secuencial la DxMx aumentó en un 26,7 por ciento. El grupo que recibió esquema continuo combinado con valerianato de estradiol más MPA (2,5 mg) evidenció un aumento de la DxMx de un 30 por ciento. Estos resultados señalan la necesidad de agregar al informe Mx los cambios observados en la DxMx como consecuencia de la HTR ya que permite al clínico adecuar los esquemas terapéuticos a la respuesta individualizada de cada mujer

Efecto reductor de la lactona sesquiterpénica "Helenalina" y sus derivados metálicos en la formación del complejo estradiol receptor en tejido tumoral mamario / Reductive effect of sesquiterpenic lactone \"Helenaline\" and its metalic derivates in complex estradiol-receptor formation in breast tumoral tissue

Dentro de la terapia sistémica empleada para el tratamiento de cáncer mamario, ha sido extensamente utilizada la quimioterapia, la cual ha sido apoyada por muy diversos compuestos en cuanto a origen y composición química. Sin embargo, todos ellos, producen diversos efectos colaterales adeversos, dignos de tomarse en cuenta. Por este hecho, precisa estudiar nuevas posibilidades en donde el fármaco aplicado, actúa selectivamente sobre célula tumoral, sin lesionar tejido sano. Para su efecto, se estudió una gamma lactona llamada "Helenalina" y sus derivados metálicos He-Co, He-Hg y He-Cu, cuya composición química les permite reaccionar con residuos -SH presentes en el receptor de la célula tumoral, los cuales al intercalarse por una reacción previa, podría modificar su composición estructural y finalemente su afinidad por la hormona. Se investigó el efecto de inhibición para la formación del complejo estradiol-receptor en el citosol de tejido tumoral mamario empleando Helenalina a 12 n M y 126 n M, obteniéndose un efecto de inhibición de 14 por ciento y 56 por ciento respectivamente. Cuando se estudió He-Co, He-Hg y He-Cu este efecto se vió aumentado, obteniéndose 11 por ciento, 10.5 por ciento y 60 por ciento con 12 n m y 44.5 por ciento, 74.5 por ciento y 86 por ciento con 126 n M respectivamente

Increased sensitivity and accumulation os estradiol in the rat oviduct during early pregnancy

We have previously reported that a single injection of estradiol-17 beta (E2) given on day 3 of pregnancy (P3) is far more effective for accelerating oviductal transport in the rat, than treatment given on day 1 (P1). In order to quantify this change, dose-response curves were established for six different doses of E2 (range 0.031 to 1.00 micrograms per animal) given on P1, P2 or P3. In addition, a possible mechanism was explored by comparing the plasmatic and oviductal levelsof E2 between 30 and 180 min following treatment with E2 on P1 or P3. As the interval from ovulation to treatment was increased, the transport of a larger number of embryos was accelerated and a smaller dose was required. The minimal effective dose decreased 30-fold from P1 to P3, the oviducts accumulated 20 percent to 90 percent more E2 on P3 than on P1, tissuelevels were 6- to 48-fold higher than plasma levels and the latter did not differ between P1 and P3. It is concluded that the oviduct exhibits increased sensitivity and responsiveness to E2 on P3 and this isassociated with greater accumulation of the hormone in the organ, not attributable to higher E2 plasma levels