Relationship between follicular dynamics and oocyte maturation during in vitro culture as a non-invasive sign of caprine oocyte meiotic competence.

The search for non-invasive signs of oocyte meiotic competence is very important for the development of in vitro follicle culture (IVFC) systems. The aims of the present study were: (1) to investigate the effect of in vitro maturation (IVM) of in vivo grown goat COCs, in group or individually, on oocyte chromatin configuration (Experiment 1), and (2) the influence of IVFC period (12 vs. 18 days) on the ability of the oocyte to resume meiosis immediately after IVFC (before in vitro maturation; IVM), or after IVM (Experiment 2). In experiment 1, in vivo grown cumulus-oocyte complexes (COCs) were submitted to IVM in groups (10 COCs/100 µL-drop) or individually (1 COC/10 µL-drop), and chromatin configuration was assessed. In experiment 2, isolated follicles were individually cultured for 12 or 18 days, and submitted to individual IVM afterwards. The following end points were evaluated: follicular growth and morphology, oocyte diameter, viability and chromatin configuration, as well as individual follicular estradiol production. Similar maturation rates were obtained between in vivo grown COCs matured individually and in groups (66.7% vs. 63.6%, respectively) (Experiment 1). Only after 18 days of IVFC, oocytes were able to grow during IVM, reaching a mean oocyte diameter of 119 µm. Also, this treatment produced the highest rate of metaphase II oocytes (46.2% out of the total number of cultured follicles). Finally, it was observed that follicles with a daily growth rate >7.1 µm/day (fast-growing) and that reached at least 600 µm in diameter, were more likely (P < 0.05) to produce oocytes capable of attaining MII. In conclusion, caprine oocytes can be individually matured in vitro, as efficiently as in groups. This result was essential to pair in vitro follicle development and in vitro oocyte maturation with specific individual follicles. Using this approach, it was possible to establish non-invasive signs for the efficiency of IVFC based on follicle daily growth rate and diameter, and oocyte diameter: follicle daily growth >7 µm, follicle diameter of at least 600 µm, and oocyte diameter ≥120 µm. In addition, 18 days seems to be the most suitable culture time for caprine early antral follicles.

Refining insulin concentrations in culture medium containing growth factors BMP15 and GDF9: An in vitro study of the effects on follicle development of goats.

The aim of the present study was to investigate the influence of two insulin concentrations (10ng/mL and 10µg/mL) combined or in the absence of BMP15 and/or GDF9, on the in vitro survival and development of preantral follicles of goat ovarian tissue. Ovarian slices from the same goat ovary pair were randomly assigned to a non-cultured control treatment or to be in vitro cultured for 1 or 7days in α-MEM containing 10ng/mL (Low) or 10µg/mL (High) of insulin in the absence or presence of BMP15 and/or GDF9. With the low insulin treatment, there was a greater percentage of normal follicles than with the high insulin treatment. The addition of BMP15 alone or in association with GDF9 to the medium containing low insulin resulted in a lesser percentage of normal follicles (P<0.05). The addition of BMP15 and GDF9 separately or in combination with the high insulin concentration enhanced the percentage of normal follicles. On day 7 of culture, the use of medium containing low insulin alone or high insulin supplemented with BMP15 and BMP15+GDF9 resulted in a greater percentage of secondary follicles than the non-cultured control, although follicles cultured with low insulin were smaller than those from the control group and had greater rates of oxidative stress. In conclusion, in the presence of physiological concentrations of insulin (10ng/mL), medium supplementation with GDF9 and BMP15 alone or in combination is unnecessary for normal follicle development in vitro.

Caprine ovarian follicle requirements differ between preantral and early antral stages after IVC in medium supplemented with GH and VEGF alone or in combination.

The aim of the present study was to evaluate the effect of growth hormone (GH) and vascular endothelial growth factor (VEGF) added alone, sequentially or in combination, in the presence of insulin at physiological concentration (10 ng/mL) on the IVC of two different follicular categories: preantral (experiment 1; Exp.1) and early antral (experiment 2; Exp.2). Isolated follicles were individually cultured for 24 (Exp.1) and 18 days (Exp.2) in the following treatments: αMEM+ (Control), or Control medium supplemented with 50 ng/mL GH (GH), 100 ng/mL VEGF (VEGF), the combination of both (GH + VEGF), GH during the first 12 days and VEGF from Day 12 until the end of the culture (GH/VEGF) and vice versa (VEGF/GH). At the end of the culture, cumulus-oocyte complexes from in vitro-grown follicles were recovered and subjected to IVM. The following end points were evaluated: Follicle morphology, growth rates and antrum formation, production of estradiol, progesterone and testosterone, oocyte viability and meiotic stage, as well as relative expression of LHR, Amh, HAS2, PTGS2, CYP17, CYP19A1, and 3ßHSD. A considerable amount of viable fully grown oocytes were recovered after the IVC of early antral follicles in all treatments. Nevertheless, the GH treatment presented the highest percentage of fully grown oocytes (60%), mean oocyte diameter (117.74 ± 2.61 µm), and meiotic resumption (50%). Furthermore, GH treatment produced higher (P < 0.05) rates of metaphase II oocytes than all the other treatments, and similar LHR, Amh, and PTGS2 transcript levels to in vivo. Contrary to early antral follicles, preantral follicles were not affected by medium supplementation. In conclusion, the addition of GH to a culture medium containing physiological concentrations of insulin improves oocyte growth and maturation after the IVC of goat early antral follicles.

Evaluation of different doses of mashua (Tropaeolum tuberosum) on the reduction of sperm production, motility and morphology in adult male rats.

Mashua is an edible-tuber crop that grows in the Andean region. Folk medicine describes the use of mashua to reduce reproductive function in men. The present study aimed: (i) to determine whether different doses of mashua (0.01, 0.1, 1 and 2 g kg(-1)) produced a dose-response reduction on sperm production and quality; and, (ii) to determine whether these anti-reproductive effects of mashua can be reversible after cessation of treatment (12 and 24 days of recovery time). Mashua-treated rats showed lower values of daily sperm production, epididymal and vas deferens sperm count and sperm motility; meanwhile, mashua increased the percentage of abnormal sperm morphology and epididymal sperm transit rate. The following variables follow a dose-response effect: sperm number in vas deferens, sperm motility and sperm transit rate. In addition, it was demonstrated that the reduction in reproduction function in male rats treated with mashua was reversible after 24 days of recovery time. Finally, lower doses mashua reduces sperm number and quality (motility and morphology), and these adverse effects on male reproductive system may be reversible after 24 days after cessation of the treatment.

Effect of red maca (Lepidium meyenii) on prostate zinc levels in rats with testosterone-induced prostatic hyperplasia.

Lepidium meyenii (maca) is a plant that grows exclusively above 4000 m in the Peruvian central Andes. Red maca (RM) extract significantly reduced prostate size in rats with benign prostatic hyperplasia (BPH) induced by testosterone enanthate (TE). Zinc is an important regulator of prostate function. This study aimed to determine the effect of RM on prostate zinc levels in rats with BPH induced by TE. Also, the study attempted to determine the best marker for the effect of RM on sex accessory glands. Rats treated with RM extract from day 1 to day 14 reversed the effect of TE administration on prostate weight and zinc levels. However, RM administered from day 7 to day 14 did not reduce the effect of TE on all studied variables. Finasteride (FN) reduced prostate, seminal vesicle and preputial gland weights in rats treated with TE. Although RM and FN reduced prostate zinc levels, the greatest effect was observed in TE-treated rats with RM from day 1 to day 14. In addition, prostate weight and zinc levels showed the higher diagnosis values than preputial and seminal vesicle weights. In conclusion, RM administered from day 1 to day 14 reduced prostate size and zinc levels in rats where prostatic hyperplasia was induced with TE. Also, this experimental model could be used as accurately assay to determine the effect of maca obtained under different conditions and/or the effect of different products based on maca.

Vanadium recovery from oil fly ash by leaching, precipitation and solvent extraction processes.

In order to reduce the environmental impact due to land disposal of oil fly ash from power plants and to valorize this waste material, the removal of vanadium was investigated using leaching processes (acidic and alkaline treatments), followed by a second step of metal recovery from leachates involving either solvent extraction or selective precipitation. Despite a lower leaching efficiency (compared to sulfuric acid), sodium hydroxide was selected for vanadium leaching since it is more selective for vanadium (versus other transition metals). Precipitation was preferred to solvent extraction for the second step in the treatment since: (a) it is more selective; enabling complete recovery of vanadate from the leachate in the form of pure ammonium vanadate; and (b) stripping of the loaded organic phase (in the solvent extraction process) was not efficient. Precipitation was performed in a two-step procedure: (a) aluminum was first precipitated at pH 8; (b) then ammonium chloride was added at pH 5 to bring about vanadium precipitation.

[Validation of the cedars-sinai score in the prediction of rebleeding and mortality in non variceal upper gastrointestinal hemorrhage]. / Validación del puntaje de cedars-sinai en la predicción de resangrado y mortalidad en hemorragia digestiva alta no variceal.

INTRODUCTION: Upper gastrointestinal hemorrhage is a serious health problem with mortality rates that have remained unchanged for the last years in spite of improvements in management. OBJECTIVES: Validating the Cedars Sinai score for predicting rebleeding and mortality at the Arzobispo Loayza National Hospital. MATERIAL AND METHODS: Prospective longitudinal study in patients with upper gastrointestinal hemorrhage, collecting demographic, clinical and endoscopic data. The Cedars-Sinai score was applied in order to classify patients: low risk, medium risk and high risk. The final results were rebleeding and mortality. Calibration was done by X2, and the discrimination of the score was assessed by analysis of ROC curves. An area below the curve greater than 0.5 was considered good discrimination. RESULTS: We evaluated 143 patients, predominantly male (54.5%), the median age was 57.8. Melena (48.95%) and hematemesis (16.08) were the most frequent symptoms. Out of the total number of patients, 36.3% was taking some type of medication; NSAID's were taken by 55.7% and ACEI's by 25%. Associated comorbidity was found in 56.6%. The following diagnoses were made: gastric ulcer (32.15%), duodenal ulcer (30.75%), erosive hemorrhagic gastritis (20.3%), advanced gastric neoplasia (6.3%) and vascular lesions (4.9%). Rebleeding and mortality were 8.4% and 4.2%, respectively. The low risk group did not show any adverse events; the medium risk group, 7.25% and the high risk group, 13.2%. The areas under the curve were for rebleeding (0.79+/-0.13; p=0.01) and mortality (0.73+/-0.12; p=0.01) and considering both (0.66+/-0.12; p=0,034). CONCLUSIONS: This score is applicable in our environment and it is suitable for predicting rebleeding and mortality in patients with upper gastrointestinal hemorrhage, especially for mortality.

Validación del puntaje de Cedars-Sinai en la predicción de resangrado y mortalidad en la hemorragia digestiva alta no variceal / Validation of the score of Cedars-Sinai in the prediction of re-hemorrhage and mortality in the gigestive hemorrhage high nonvariceal

Introducción: La hemorragia digestiva alta es un serio problema de salud, con índices de mortalidad no modificados durante los últimos años, a pesar de mejoría en el manejo. Objetivo: Validar el puntaje de Cedars-Sinai, para predicción de resangrado y mortalidad en el Hospital Nacional Arzobispo Loayza. Material y Métodos: Estudio prospectivo longitudinal, en pacientes con hemorragia digestiva alta; recolectando datos demográficos, clínicos y endoscópicos. Se aplicó el puntaje de Cedars-Sinai para clasificar a los pacientes: de bajo riesgo, riesgo medio y riesgo alto. Los resultados finales considerados fueron resangrado y mortalidad. La calibración se efectuó mediante Chi cuadrado, y la discriminación de puntaje se evaluó por análisis de curvas operativas del receptor (ROC). Se consideró buena discriminación un área bajo la curva mayor de 0.5. Resultados: Se evaluaron 143 pacientes, predominando el sexo masculino (54.5 por ciento), la edad media fue 57.8 años. La melena (48.95 por ciento) y la hematemesis (16.08), fueron los síntomas más frecuentes. Del total, 36,3 por ciento tomaban algún medicamento; correspondiendo AINEs a 55.7 por ciento e Inhibidores de la enzima covertidora de la angiotensina (IECA), el 25 por ciento. El 56.6 por ciento presentó comorbilidad asociada. Se diagnosticó úlcera gástrica (32.15 por ciento), úlcera duodenal (30.75 por ciento), gastritis erosiva-hemorrágica (20.3 por ciento), neoplasia gástrica avanzada (6.3 por ciento) y lesiones vasculares (4.9 por ciento). El resangrado y la mortalidad fueron 8.4 por ciento y 4.2 por ciento, respectivamente. El grupo de bajo riesgo no presentó eventos adversos; el de riesgo medio 7.25 por ciento y el de riesgo elevado, 13.2 por ciento. Las áreas bajo la curva fueron para resangrado (0.79 más menos 0.13; p=0.01) y mortalidad (0.73 más menos 0.12; p=0.01) y considerando ambos (0.66 más menos 0.12; p=0,034). Conclusiones: Este puntaje es aplicable en nuestro medio; adecuado para...

Introduction: Upper gastrointestinal hemorrhage is a serious health problemwith mortality rates that have remained unchanged for the last years in spite ofimprovements in management. OBJECTIVES. Validating the Cedars Sinai score for predicting rebleeding and mortality at the Arzobispo Loayza National Hospital. MATERIAL AND METHODS. Prospective longitudinal study in patients with upper gastrointestinal hemorrhage, collecting demographic, clinical and endoscopie data. The Cedars-Sinai score was applied in order to classify patients: low risk, medium risk and high risk. The final results were rebleeding and mortality. Calibration was done by X2, and the discrimination of the score was assessed by analysis of ROC curves.An area below the curve greater than 0.5 was considered good discrimination.RESULTS: We evaluated 143 patients, predominantly male (54.5 percent), the median age was 57.8. Melena (48.95 percent) and hematemesis (16.08) were the most frequent symptoms.Out of the total number of patients, 36.3 percent was taking some type of medication; NSAID's were taken by 55.7 percent and ACEI's by 25 percent. Associated comorbidity was found in 56.6 percent. The following diagnoses were made: gastric ulcer (32.15 percent), duodenal ulcer (30.75 percent), erosive hemorrhagie gastritis (20.3 percent), advanced gastric neoplasia (6.3 percent) and vascular lesions (4.9 percent). Rebleeding and mortality were 8.4 percent and 4.2 percent, respectively. The low risk group did not show any adverse events; the medium risk group, 7.25 percent and the high risk group, 13.2 percent. The areas under the curve were for rebleeding (0.79:1:0.13;p=0.01) and mortality (0.73:1:0.12;p=0.01) and considering both (0.66:1:0.12; p=0,034). CONCLUSIONS. This score is applicable in our environment and it is suitable for predicting rebleeding and mortality in patients with upper gastrointestinal hemorrhage, especially for mortality.

Copper sorption by chitosan in the presence of citrate ions: influence of metal speciation on sorption mechanism and uptake capacities.

The presence of organic ligands in a solution containing metal ions modifies metal speciation, which in turn changes the sorption mechanism, optimum pH range and maximum sorption capacity. The present work investigates the sorption of copper by chitosan in the presence of citrate at different metal/ligand ratios. Copper uptake in acidic solution takes place through electrostatic attraction between the protonated amine groups of chitosan and anionic copper-citrate complexes (mainly Cu(OH)L2- but also a small fraction of CuL-). Sorption was negligible below pH 3 due to competition from dissociated anionic ligand and counter ions brought about by dissociation of the acid used for pH control. Actually, copper sorption begins to be significant when the fraction of anionic copper-complexes exceeds that of anionic copper-free ligand. So sorption capacity strongly increases up to pH 4.5-5.5. Above pH 5.5, the progressive decrease of amine protonation leads to a linear decrease in sorption capacity. An excess of ligand leads to an increase in the fraction of free dissociated (anionic) ligand that may compete for electrostatic attraction on protonated amine groups and therefore leads to a decrease in sorption capacities.