Impact of prolonged oocyte incubation time before vitrification on oocyte survival, embryo formation, and embryo quality in mice.

Oocyte incubation time before freezing is one of the factors affecting oocyte vitrification. In the assisted reproductive technology (ART) clinics, it is sometimes decided to perform oocyte vitrification after a long period of incubation time due to various conditions, such as inability to collect semen samples, unsuccessful urological interventions (PESA, TESE, etc.), or unexpected conditions. A time factor of up to 6 h has been studied in the available reports. Therefore, this study was designed to evaluate oocyte incubation time before freezing at 0, 6, 12, 18, and 24 h after retrieval. Metaphase II (MII) oocytes were obtained from NMRI female mice after being randomly divided into the five groups of 0, 6, 12, 18, and 24 h of freezing via hormonal stimulation following retrieval and entered into the vitrification-warming process. The thawed oocytes were evaluated according to the survival criteria and then inseminated with the sperms of male mice for in vitro fertilization. The next day, the embryo formation rate and embryo quality were assessed. Our results demonstrated that even after 24 h of incubation, the survival rate of oocytes was 51.35% with the embryo formation rate of 73.21%. However, the survival and embryo formation rates significantly decreased within 12, 18, and 24 h after retrieval compared to the groups vitrified at 0 h. The embryo quality was significantly reduced by vitrification at 0 to 24 h after retrieval. According to our data, although a prolonged incubation time before freezing reduced the survival rate, there was still a chance for oocytes to stay alive with acceptable embryo formation and quality rates after vitrification warming of oocytes.

The Effects of Testosterone on Oxidative Stress Markers in Mice with Spinal Cord Injuries.

BACKGROUND: Spinal cord injury (SCI) causes infertility in male patients through erectile dysfunction, ejaculatory dysfunction, semen and hormone abnormalities. Oxidative stress (OS) is involved in poor semen quality and subsequent infertility in males with SCI. The aim of this study is to examine the effects of SCI on the level of testosterone hormone. MATERIALS AND METHODS: In this experimental study, we evaluated the effects of exogenous testosterone on the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) as well as the levels of malondialdehyde (MDA) and protein carbonylation (PCO), as markers of OS, in 10 groups of SCI mice. Total antioxidant capacity (TAC) was determined using the 2,29-azinobis-(3-ethylbenzothiazoline- 6-sulfonic acid) (ABTS) radical cation assay. RESULTS: Exogenous testosterone administration in mice with SCI significantly reduced SOD and GPx enzyme activities and MDA level. There was no significant decrease in PCO content. In addition, TAC remarkably increased in the sham and SCI groups not treated with testosterone but remained unchanged in all other experimental groups. Exogenous testosterone also reduced serum testosterone levels in all groups except the positive control group. CONCLUSION: Our cumulative data indicated that SCI could cause sterility by disturbing the plasmatic testosterone balance. The normal level of endogenous testosterone was not completely restored by exogenous testosterone administration.

The effect of time and temperature on viability and performance of Langerhans islets separated from Balb/c mouse after death.

BACKGROUND: Tissue transplantation plays a pivotal role in the treatment of diseases. Pancreatic beta cell transplantation is the best way to obtain normal blood glucose in patients with diabetes type 1. However, it is not clear how long endocrine pancreas cells can be used for transplantation after the donor's death. The present study was conducted to analyze the performance and viability of pancreatic islet cells after death. MATERIALS AND METHODS: Pancreas was separated from Balb/c mice at different times (0, 1, 4, 6, 12, and 24 h after death) at temperatures of 4°C and 23°C, and was cultured in Roswell_Park_Memorial_Institute (RPMI) 1640. Insulin shock, MTT assay, aldehyde fuchsin staining, dithizone staining, and florescence microscopy methods were applied to analyze the performance of beta cells, cell viability, islets' diagnosis, islet cells' diagnosis, and viable and necrotic cells diagnosis, respectively. RESULTS: Islets of Langerhans and beta cells were diagnosed. By increasing the temperature and time, the viability and performance of beta cells decreased significantly (P < 0.05). CONCLUSION: The best condition for keeping the islets of Langerhans in terms of viability and performance is 4 h after death at temperature of 4°C.

Role of phosphodiesterase inhibitor Ibudilast in morphine-induced hippocampal injury.

BACKGROUND: Opioid drugs are used in the treatment of acute post-surgical pain and chronic pain, such as those associated with cancer. Opioid used is associated with complications such as analgesic tolerance, dependence and opioid abuse. The molecular mechanisms of unwanted opioid responses are varied but recent advances have highlighted elevations in pro-inflammatory cytokines and pro-inflammatory glial following chronic administration of morphine. In this study we investigated the neurodegenerative effects of morphine through its effects on Toll-Like Receptor 4 (TLR4) in the male rat hippocampus and evaluated the level of Interleukin-1 beta (IL-1ß). Then we compared the difference between inhibitory effects on mu opioid receptors (by ß-Funaltrexamine, ß-FNA) and TLR4 (by Ibudilast). Subsequently, we assessed the amount of IL-1ß and the number of granular cells in male rat hippocampus. METHODS: Adult male rats (n=24) were treated with sucrose, morphine, Ibudilast (7.5 mg/kg) and ß-FNA (20 mg/kg) for 30 days. Their brains were isolated and hemisected with one hippocampus for granular cell and the other used for IL-1 ß immunoblotting. RESULTS: Data showed that Ibudilast suppresses IL-1 ß expression significantly more than ß-FNA. The granular cell count displayed significant differences. CONCLUSIONS: Our results suggested that Ibudilast can be used for controlling and treatment of morphine-induced CNS inflammations or traumatic conditions.

Contemporary staining of arterial system with cadaver fixation: an experimental study / Tinción contemporánea del sistema arterial con fijación cadavérica: un estudio experimental

Although many methods are investigated, cadavers have unique role for education of gross anatomy. This study was performed to make cadaveric based gross anatomy more enjoyable and decrease the time needed for achievement of arterial system for teachers and learners. The adult rabbits in control (n=3) and experimental groups (n=9) sacrificed following deep anesthesia. The samples were fixed by injection of fixative solution to the left ventricle and the corps dropped to the traditional fixative liquid for at least 1 month. The injected fixative solution of experimental group contained stained human red blood cells. The red blood cells stained by eosin (0.25 percent in saline) for 30 minutes and injected between two injections of fixative. After passing the period of fixation, the samples were dissected and arterial system in the Neck, Trunk and extremities were recognized. The degree of staining of the arteries categorized as low, medium and high. The data of staining of arteries were analyzed by chi square using SPSS 16.0 for Windows XP (SPSS Inc., Chicago, IL). P value< 0.05 was considered significant. In almost all of the experimental samples, axillary and femoral arteries were stained (P= 0.005). The arteries of the neck (carotids), the wall of thorax (Intercostals), arch of aorta, abdominal arteries (renal, mesenteric) and iliac arteries stained (p=0.000). This study presents inexpensive and readily used protocol for making easier learning and teaching of gross anatomy by differentiating arteries from their associated nerves and veins in cadavers...

Aunque muchos métodos de enseñanaza se han investigado, los cadáveres tienen un papel único para la enseñanza de anatomía macroscópica. Este estudio se realizó para hacer de la anatomía basada en el cadáver más agradable y reducir el tiempo necesario para el aprendizaje del sistema arterial de profesores y alumnos. Dos grupos de conejos adultos, uno control (n = 3) y otro experimental (n = 9) fueron sacrificados después de una anestesia profunda. Las muestras fueron fijadas mediante la inyección de solución en el ventrículo izquierdo manteniéndose los cuerpos en líquido fijador durante al menos 1 mes. La solución fijadorea inyectada en el grupo experimental contenía glóbulos rojos (GR)humanos teñidos. Los GR se tiñeron con eosina (0,25 por ciento en solución salina) durante 30 minutos y se inyectaron entre dos inyecciones de fijador. Después del período de fijación, las muestras fueron disecadas, y se reconoció el sistema arterial en el cuello, tronco y miembros. El grado de tinción de las arterias fue clasificado como bajo, medio y alto. Fue utilizado el test de Chi cuadrado con el programa SPSS 16.0 para Windows XP (SPSS Inc., Chicago, IL). Un valor P < de 0,05 fue considerado significativo. En casi la totalidad de las muestras experimentales, se tiñeron las arterias axilar y femoral (P = 0,005). Se tiñeron las arterias del cuello (carótidas), de la pared del tórax (intercostales), arco aórtico, arterias abdominales (renal y mesentérica) y arterias ilíacas (p = 0,000). Este estudio presenta un protocolo realizable de bajo costo que permite hacer más fácil el aprendizaje y la enseñanza de la anatomía macroscópica, permitiendo diferenciar las arterias de los nervios y venas asociadas...

Growth pattern in 7-12 years old Arak children (central Iran) in comparison with other ethnic subgroups of Iran.

BACKGROUND: growth is a remarkable index of health and can be influenced by genetic and environment conditions. The pattern of growth is unique for every nation and worldwide studies have demonstrated separate national standards. In the case of Iran, there is not enough information in this field. METHODS: This study was undertaken on 7-12 years old children from Arak. The data for each individual such as age, height, weight and body mass index (BMI) were recorded. Differences in the data between two sexes were tested by means of the paired sample t- test and the mean BMI was compared with sex- and age-specific reference values from the National Center for Health Statistics of the Centers for Disease Control (CDC) 2000 growth chart using independent sample t-tests. Levels of P < 0.05, P < 0.01 and P < 0.001 were recorded as significant. RESULTS: In this study, males were significantly taller and heavier than females at the same age (P < or = 0.01) except for length at age 10 and weight at age 12. The BMI curves were between 50th and 25th percentiles of CDC. CONCLUSIONS: The present study shows the effect of socioeconomic background that had been also considered in other studies in Iran. There is a need for ethnic specific growth charts and BMI cut-off points for underweight, overweight and obesity in children of each ethnic subgroup of Iranian population.

Age at menarche and related factors in girlsof urban areas of Markazi (Central) province of Iran / Edad de la menarquia y factores relacionados en las niñas de zonas urbanas de la provincia Markazi (Central) de Irán

The age at menarche is an indicator of racial, geographical and nutritional patterns of different societies. This cross-sectional study conducted on 1223 girls aged 14-20 in the Markazi (Central) Province, Iran in 2010. In this research, the age at menarche, as the main variable, was determined for each city and compared between different cities. The statistical analyses included c2 test for determining statistically significant differences, variance analysis for determining the difference between groups, and Pearson correlation coefficient for determining the relationship between variables. The average age of first menstruation was 13.21+/-1.33 years in the Markazi Province, with no significant difference among the cities of the province. This age is inversely related to body mass index (BMI), nutritional status and weight. The pattern of distribution of BMI is similar for all the cities in the province. Although, the average age at menarche in the Markazi (central) Province, a cold area, is higher compared to warmer regions in Iran, it may be more influenced by race than by the climate. Considering the lower age at menarche in Markazi Province compared to previous similar studies, it may be stated that social well-being and nutritional standards have improved in the province.

La edad de la menarquia es un indicador de patrones raciales, geográficos y nutricionales de distintas sociedades. Este estudio transversal fue realizado el 2010 en 1223 niñas entre 14 y 20 años en la Provincia Markazi (Central), Irán. En esta investigación, la edad de la menarquia, como la variable principal, se determinaron para cada ciudad y en comparación entre las diferentes ciudades. Los análisis estadísticos incluyeron la prueba c2 para determinar diferencias estadísticamente significativas, el análisis de varianza para determinar la diferencia entre los grupos, y el coeficiente de correlación de Pearson para determinar la relación entre las variables. El promedio de edad de la primera menstruación fue 13,21+/-1,33 años en la provincia de Markazi, sin diferencias significativas entre las ciudades de la provincia. Esta edad es inversamente proporcional al índice de masa corporal (IMC), el estado nutricional y el peso. El patrón de distribución del IMC es similar para todas las ciudades de la provincia. Aunque la edad media de la menarquia en la Provincia Markazi (central), una zona de clima frío, es mayor en comparación con las regiones más cálidas en Irán, puede estar más influenciadas por la raza que por el clima. Teniendo en cuenta la baja edad de menarquia en la Provincia Markazi en comparación con estudios similares anteriores, se puede afirmar que el bienestar social y los niveles de nutrición han mejorado en la provincia.

The enhancing effects of alcoholic extract of Nigella sativa seed on fertility potential, plasma gonadotropins and testosterone in male rats.

BACKGROUND: The task force on plants for fertility regulation in men continued with its program to identify novel prototypes in plants alleged to have fertility regulating properties. Nigella Sativa seeds are frequently used in folk medicine in the Middle East and some Asian countries for the promotion of good health and treatment of many ailments. OBJECTIVE: To evaluated the role of alcoholic extract of Nigella sativa on fertility potential, Pituitary-testicular axis hormones and Testosterone in male rats. MATERIALS AND METHODS: 24 male rats were randomly divided into 3 groups; control, group A and group B, each group comprising of 8 rats. Animals in control group received 1 ml of normal saline and treatment groups (A and B) received (gavage) graded doses of 200 and 400 mg/kg body weight of alcoholic extract of Nigella sativa seeds on a daily basis for 60 days. At the end of treatment period, fertility parameters such as body and reproductive organs weight, sperm motility, viability and count, epididymal sperm reserve (ESR), daily sperm production (DSP), blood testosterone concentration, Gonadotropins levels and fertility index were measured. RESULTS: There was a significant difference in testes and epididymidis weight, sperm count, ESR, DSP, blood testosterone concentration, LH and fertility index in both the lower dose group and the higher group as compared to the control group. CONCLUSION: The results of this study showed that alcoholic extract of Nigella sativa seed especially in higher doses could increase fertility potential, LH and testosterone concentration in male rats.

Effect of exposure to extremely low electro-magnetic field during prenatal period on mice spleen.

Total body weight of newborns, the volume of spleen, and the number of megakaryocytes decreased following the exposure to ELF-MF (6 x 10(-3) T and 50 Hz) at 1-5, 6-10, 11-15, and 16-20 days of pregnancy of mice. The complete period of gestation was sensitive to ELF-MF exposure; the initial days were more prone to exposure. The results suggest that the use of ELF-MF producing instruments should be limited during gestation.

Mind map teaching of gross anatomy is sex dependent / Mapa conceptual en la enseñanza de la anatomía macroscópica es dependiente del sexo

The aim of this study was to evaluate the effectiveness of using Mind Maps in teaching anatomy and compare it with traditional based slide. This cross-sectional research was carried out in 2009 on 79 second year medical undergraduate students at Ahvaz Jondishapour University of Medical sciences. Four class lectures on the topics of head and neck osteology were taught. The lectures were slide based designed that were chosen from the chapters in the anatomy textbooks and presented by means of a video projection power point system. The students divided randomly into two groups; one group educated by traditional slides while the other by mind map based designed ones. At the end of the term, a multiple-choice involved forty test was undertaken from two groups. The data was investigated using analysis of variance (ANOVA) and paired T test (p<0.05). The mean scores of females in mind map group were higher than that for traditional one (p<0.04), while there were no significant differences in the mean scores of the men of two groups. The mean scores of females were significantly higher than the males in mind map group (p<0.02) and although the mean scores of females in traditional group was higher it was not significant. This paper has shown the efficacy of using mind maps for teaching gross anatomy as a study aid, even when use has been limited to a single exposure. The study found sex-dependent of learning, suggesting that sex may determine the optimal neural network for designing teaching protocols. Indeed, for females, that have obtained better scores, the use of bilateral neural network seems to facilitate the learning process by mind map teaching of anatomy.

El objetivo de este estudio fue evaluar la efectividad del uso de mapas conceptuales en la enseñanza de la anatomía y compararla con la tradicional basada en diapositivas. Esta investigación de corte transversal se llevó a cabo en el año 2009 en 79 estudiantes de pregrado del Segundo Año de Medicina en la Universidad de Ciencias Médicas Ahvaz Jondishapour. Fueron analizadas cuatro clases conferenciales en los temas de cabeza y cuello óseo. Las conferencias diseñadas se basaron en el uso de diapositivas las que fueron seleccionadas desde los capítulos de libros de Anatomía y se presentaron por medio de proyección de videos utilizando el sistema power point. Los estudiantes fueron divididos aleatoriamente en dos grupos: un grupo educado por diapositivas tradicionales mientras que el otro se basó en el diseño de mapas mentales. Al final del ciclo de conferencias, una prueba de opción múltiple de cuarenta preguntas se aplicó en ambos grupos. Los datos fueron investigados mediante el análisis de varianza (ANOVA) y prueba de la t de Student (p<0,05). Las puntuaciones medias de las mujeres en el grupo de mapa mental fueron mayores que las del grupo diapositivas (p<0,04), mientras que no hubo diferencias significativas en las puntuaciones medias de los hombres de ambos grupos. Las puntuaciones medias de las mujeres fueron significativamente mayores que los hombres en el grupo de mapa mental (p <0,02) y aunque las puntuaciones medias de las mujeres en el grupo diapositivas fue más alto, no fue estadísticamente significativo. En este trabajo se ha demostrado la eficacia del uso de mapas mentales para la enseñanza de anatomía como una ayuda al estudio, incluso cuando el uso se ha limitado a una sola exposición. El estudio encontró que el aprendizaje depende del sexo, lo que sugiere que el sexo puede determinar una red neuronal para el diseño de protocolos óptimos de enseñanza. En efecto, para las mujeres, que tienen mejores calificaciones, el uso de redes neuronales bi...

Sildenafil Effect on Nitric Oxide Secretion by Normal Human Endometrial Epithelial Cells Cultured In vitro.

BACKGROUND: Sildenafil is a selective inhibitor of cyclic-guanosine monphosphat-specific phosphodiesterase type 5. It increases intracellular nitric oxide (NO) production in some cells. There are reports on its positive effect on uterine circulation, endometrial thickness, and infertility improvement. Endometrial epithelial cells (EEC) play an important role in embryo attachment and implantation. The present work investigates the effect of sildenafil on human EEC and their NO secretion in vitro. MATERIALS AND METHODS: In this experimental in vitro study, endometrial biopsies (n=10) were washed in a phosphate buffered solution (PBS) and digested with collagenase I (2 mg/ml in DMEM/ F12 medium) at 37°C for 90 minutes. Epithelial glands were collected by sequential filtration through nylon meshes (70 and 40 µm pores), respectively. Epithelial glands were then treated with trypsin to obtain individual cells. The cells were counted and divided into four groups: control and 1, 10, and 20 µM sildenafil concentrations. Cells were cultured for 15 days at 37ºC and 5% CO2; the media were changed every 3 days, and their supernatants were collected for the NO assay. NO was measured by standard Greiss methods. Data were analyzed by one way ANOVA. RESULTS: There was no significant difference between groups in cell count and NO secretion, but the level of NO increased slightly in the experimental groups. The 10 µM dose showed the highest cell count. EEC morphology changed into long spindle cells in the case groups. CONCLUSION: Sildenafil (1, 10, and 20 µM) showed a mild proliferative effect on human EEC numbers, but no significant change was seen in NO production.