Effect of season on bovine seminal plasma proteins in Thailand.

Although season has been shown to affect bull sperm quality and fertility in some studies, the effect of season on seminal plasma proteins has not been examined. In the present study, seminal plasma proteins were analysed by Fast Protein Liquid Chromatography (FPLC), to separate the phosphorylcholine-binding proteins and heparin-binding proteins from the other proteins. Semen samples were collected from bulls in three seasons: winter, summer and the rainy season. Sperm quality was analysed by flow cytometry and computer assisted sperm analysis, and further aliquots of semen were used to prepare the seminal plasma for FPLC. Meteorological data were available from a location close to the bull station. There were slight differences in sperm kinematics between seasons, but other parameters of sperm quality were not different. Minor differences in the phosphorylcholine-binding proteins were detected according to season, being lower in summer than in winter or in the rainy season, although there were no changes in the heparin-binding proteins. Temperature, humidity and rainfall differed between winter and the rainy season, but no differences were observed between summer and the rainy season except in the temperature humidity index (THI). However, the THI was above the threshold indicative of heat stress in all seasons, which could explain why few seasonal differences in protein composition were detected in this study. Alternatively, the bulls could have been well-adapted to heat stress. In conclusion, there were only slight differences in bull sperm quality and seminal plasma proteins between seasons during this study.

Effects of di(2-etilhexil) phthalate on human umbilical artery.

Di(2-etilhexil) phthalate (DEHP) is a compound used in plastic materials, which has endocrine disrupting properties. The human DEHP exposure depend on the use of plastics in toys, medical devices and food and beverage containers. The DEHP effects were studied in some physiological systems; nevertheless, the actions in human arteries were never described. We analysed the DEHP effect on endothelium denuded human umbilical artery (HUA), an important artery to ensure gases and nutrients exchange with fetus. We assessed DEHP short-term effects on contractility, occurring few minutes after DEHP is in contact with HUA in the organ bath receptacles. The long-term effects on HUA, observed after 24 h in presence of DEHP, were assessed in the organ bath system, and also through the analysis of receptors expression (5-HT2A and H1) and of cellular viability, by using HUA smooth muscle cells. DEHP (1 nM-100 µM) induced a short-term relaxing effect on HUA contracted by 5-HT, histamine or KCl. DEHP long-term exposure of arteries (1 nM, 10 µM and 100 µM) reduced its own relaxant effect on HUA contracted by 5-HT and histamine and, precisely, 24 h exposure to DEHP 1 nM reverted the relaxant effect on 5-HT contractility. Long-term exposure at more than 10 nM of DEHP decreased 5HT2A receptors expression. In conclusion, DEHP short-term exposition elicit vasodilation of HUA contracted by different agents. DEHP long-term exposition reduced the expression of 5HT2A receptors. The DEHP long-term exposition decrease the short-term relaxant effect and, at low concentrations can increase the contractile effect of 5-HT.

Effect of freezing bull semen in two non-egg yolk extenders on post-thaw sperm quality.

Traditionally, extenders for bull semen included egg yolk or milk, but recently there has been a move to avoid material of animal origin. The aim of this study was to evaluate the effects of two commercial extenders (based on soya lecithin and liposomes) on bull sperm quality after cryopreservation. Post-thaw sperm quality was evaluated by computer-assisted sperm analysis and flow cytometric assessment of membrane integrity, chromatin integrity, mitochondrial membrane potential, production of reactive oxygen species and tyrosine phosphorylation. Furthermore, an artificial insemination (AI) trial was conducted, and 56-day non-return rates were evaluated. Semen frozen in the liposome-based extender showed similar membrane integrity and higher mitochondrial membrane potential compared to those in the soya lecithin-based extender. Chromatin integrity and production of live H2 O2 + reactive oxygen species were similar in both extenders. Less superoxide was produced in the samples extended with liposome-based extender, with or without menadione stimulation. Chromatin integrity and tyrosine phosphorylation were not affected by either type of extender. No differences in 56-day non-return rate between extenders containing soya lecithin and liposomes were observed in the AI trial (66% ± 0.8 and 65% ± 0.8, respectively). In conclusion, the sperm quality of bull semen frozen in the two extenders that do not contain material of animal origin was similar, although the semen frozen in the liposome-based extender had higher mitochondrial membrane potential. Either extender could be used in situations where extenders containing material of animal origin are to be avoided.

Effect of oxygen on the microbial activities of thermophilic anaerobic biomass.

Low oxygen levels (µgO2L(-1)) in anaerobic reactors are quite common and no relevant consequences are expected. On the contrary, higher concentrations could affect the process. This work aimed to study the influence of oxygen (4.3 and 8.8mgO2L(-1), respectively) on the different microbial activities (hydrolytic, acidogenic and methanogenic) of thermophilic anaerobic biomass and on the methanogenic community structure. Batch tests in presence of oxygen were conducted using specific substrates for each biological activity and a blank (with minimum oxygen) was included. No effect of oxygen was observed on the hydrolytic and acidogenic activities. In contrast, the methane production rate decreased by 40% in all oxygenated batches and the development of active archaeal community was slower in presence of 8.8mgO2L(-1). However, despite this sensitivity of methanogens to oxygen at saturation levels, the inhibition was reversible.

Expression of angiotensin II receptors in the caprine ovary and improvement of follicular viability in vitro.

This study aimed to evaluate mRNA levels of angiotensin II (ANG II) receptors (AGTR1 and AGTR2) in caprine follicles and to investigate the influence of ANG II on the viability and in vitro growth of preantral follicles. Real-time polymerase chain reaction (PCR) was used to quantify AGTR1 and AGTR2 mRNA levels in the different follicular stages. For culture, caprine ovaries were collected, cut into 13 fragments and then either directly fixed for histological and ultrastructural analysis (fresh control) or placed in culture for 1 or 7 days in α-minumum essential medium plus (α-MEM+) with 0, 1, 5, 10, 50 or 100 ng/ml ANG II. Then, the fragments were destined to morphological, viability and ultrastructural analysis. The results showed that primordial follicles had higher levels of AGTR1 and AGTR2 mRNA than secondary follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA than their respective cumulus-oocyte complex (COCs). After 7 days of culture, ANG II (10 or 50 ng/ml) maintained the percentages of normal follicles compared with α-MEM+. Fluorescence and ultrastructural microscopy confirmed follicular integrity in ANG II (10 ng/ml). In conclusion, a high expression of AGTR1 and AGTR2 is observed in primordial follicles. Granulosa/theca cells from antral follicles had higher levels of AGTR1 mRNA. Finally, 10 ng/ml ANG II maintained the viability of caprine preantral follicles after in vitro culture.

Diabetes in the hospitalized patients with urological diseases.

BACKGROUND: Diabetes and urological diseases are widespread health problems, whose incidence increases with age. The aim of this observational, retrospective study was to analyze the particularities of urinary disorders, which appeared in patients with diabetes, admitted in a urology ward. MATERIAL AND METHODS: A total of 6910 patients admitted in "Th. Burghele" Hospital from January 2013 to July 2014 were analyzed. Only admissions in urology wards and the first hospitalization of the patient were elements that were taken into account. Data was taken from the Hipocrate medical information system and Easy Medical Pro laboratory medical software. Study variables were age and sex of patients, the main discharge diagnosis, the number of days of hospitalization and the laboratory analyses collected on the day of admission. The data of the whole lot was analyzed and then an analysis on subgroups of patients was done. RESULTS: There were 16.52% (n=1142) patients with diabetes in the total group of analyzed patients. Urinary stones were the most frequent cause of hospitalization, both in patients with diabetes and in patients without diabetes (28.5%, respectively 37.5%). The average age was 60.01 years and the mean duration of hospitalization was 6.52 days. Patients with diabetes hospitalized for urinary stones, renal cancer, and infectious pathology were significantly older than patients without diabetes. The presence of diabetes prolonged hospitalization in the case of patients with kidney stones, kidney cancer and in those with infectious pathology. The most common malignancy was bladder cancer in both groups of patients. Malignancies were more common in diabetics (19.08% vs. 15.98%) and diabetes was a risk factor for malignancy in our study. In particular, patients with diabetes had a significantly increased risk of bladder cancer. In the analyzed group, diabetes was positively associated with prostate adenoma, genital infections, and prostate infections. CONCLUSIONS: Diabetes increased the risk for certain urological diseases (bladder cancer, prostate adenoma, prostate and genital infections), it prolonged hospitalizations, and it was associated with certain features of laboratory analysis (leukocytosis, decreased glomerular filtration rate).

Sphingosine 1-phosphate promotes activation of aprine preantral follicle in vitro / Esfingosina 1-fosfato promove ativação de folículos pré-antrais de caprinos in vitro

This study describes the effect of sphingosine 1-phosphate (S1P) for development of preantral follicle, therefore the activation and follicular viability of caprine follicles cultured in vitro. Ovarian fragments were cultured for 1 or 7 days in Minimum Essential Medium with different S1P concentrations (0, 1, 10, 50, 100 or 200ng/mL). All ovarian fragments were processed for histological analysis in optical microscopy, transmission electron microscopy and fluorescence analysis. The treatment using 1ng/mL of S1P was able to maintain the percentage of normal follicles with the progression of the culture from day 1 to 7. At end of the 7-day culture period there was a significant reduction (P<0.05) in the percentage of primordial follicles in all groups treated with S1P, compared with fresh control (FC) and Control Culture (CC), which was followed by an increase of activated follicles (intermediary, primary and secondary). In addition, the culture for 7 days with media supplemented with S1P with 1ng/mL preserved the ultrastructure of organelles and kept the preantral follicular viability when evaluated by fluorescence microscopy. In conclusion, after 7 days of culture, the 1ng/mL of S1P activates the development of preantral caprine follicles, cultured in situ and maintains the oocitary and follicular viability...

Este estudo descreve o efeito da esfingosina 1-fosfato (S1P) no desenvolvimento de folículos pré-antrais, portanto da ativação e viabilidade de folículos caprinos cultivados in vitro. Fragmentos de ovários foram cultivados por um ou sete dias em meio essencial mínimo com diferentes concentrações de S1P (0, 1, 10, 50, 100 ou 200ng/mL). Os fragmentos de ovário foram processados para análise histológica em microscopia óptica, microscopia eletrônica e microscopia de fluorescência. O tratamento usando 1ng/mL de S1P foi capaz de manter a porcentagem de folículos normais durante o período de cultivo de sete dias. Ao final do período de cultivo, houve uma redução significativa (p<0,05) na porcentagem de folículos primordiais em todos os grupos tratados com S1P, comparados com os grupos controle (FC e CC), seguida por um aumento do número de folículos ativados (intermediários, primários e secundários). Adicionalmente, na cultura por sete dias com meio suplementado com S1P (1ng/mL), houve preservação da ultraestrutura das organelas e manteve-se a viabilidade dos folículos pré-antrais avaliados por microscopia de fluorescência. Em conclusão, após sete dias de cultura, o meio suplementado com 1ng/mL de S1P ativa o desenvolvimento de folículos pré-antrais de caprino, cultivados in situ e mantém as viabilidades oocitária e folicular...

Evaluation of anti-nociceptive and anti-inflammatory activity of low-level laser therapy on temporomandibular joint inflammation in rodents.

The aim of this study was to investigate the analgesic and anti-inflammatory activity of low-level laser therapy (LLLT) on the nociceptive behavioral as well as histomorphological aspects induced by injection of formalin and carrageenan into the rat temporomandibular joint. The 2.5% formalin injection (FRG group) induced behavioral responses characterized by rubbing the orofacial region and flinching the head quickly, which were quantified for 45 min. The pretreatment with systemic administration of diclofenac sodium-DFN group (10 mg/kg i.p.) as well as the irradiation with LLLT infrared (LST group, 780 nm, 70 mW, 30 s, 2.1 J, 52.5 J/cm(2), GaAlAs) significantly reduced the formalin-induced nociceptive responses. The 1% carrageenan injection (CRG group) induced inflammatory responses over the time-course of the study (24 h, and 3 and 7 days) characterized by the presence of intense inflammatory infiltrate rich in neutrophils, scanty areas of liquefactive necrosis and intense interstitial edema, extensive hemorrhagic areas, and enlargement of the joint space on the region. The DFN and LST groups showed an intensity of inflammatory response that was significantly lower than in CRG group over the time-course of the study, especially in the LST group, which showed exuberant granulation tissue with intense vascularization, and deposition of newly formed collagen fibers (3 and 7 days). It was concluded that the LLLT presented an anti-nociceptive and anti-inflammatory response on the inflammation induced in the temporomandibular joint of rodents.

Interaction between melatonin and follicle-stimulating hormone promotes in vitro development of caprine preantral follicles.

The aim of this study was to investigate the effects of melatonin and follicle-stimulating hormone (FSH) on the in vitro culture of goat preantral follicles. Ovarian fragments were cultured for 7 d in α-minimum essential medium (α-MEM(+)) containing melatonin (100, 250, 500, or 1,000 pM), FSH (50 ng/mL), or a combination of the 2 hormones and further analyzed by histology and transmission electron and fluorescent microscopy. The results showed that after 7 d of culture, tissues cultured in α-MEM(+) alone or supplemented with FSH alone, melatonin (500 and 1,000 pM), or the combination of FSH and melatonin (1,000 pM) maintained percentages of normal preantral follicles similar to the fresh control. In contrast to the noncultured tissues, the percentage of developing follicles was increased under all culture conditions after 7 d (P < 0.05). The addition of 1,000 pM melatonin associated with FSH to the culture medium increased follicular and oocyte diameters compared with α-MEM(+) alone after 7 d of culture (P < 0.05). Ultrastructural and fluorescent analyses confirmed the integrity of follicles cultured with 1,000 pM of melatonin plus FSH for 7 d. In conclusion, this study demonstrated that the interaction between melatonin and FSH maintains ultrastructural integrity and stimulates further growth of cultured caprine preantral follicles.

Frequency and mechanism of ischemic stroke associated with malignancy: a retrospective series.

BACKGROUND: Cerebrovascular disease is the second most common non-metastatic neurological disorder in oncological patients. The aims of this study were to determine the prevalence of cancer in stroke patients and to compare mechanisms of stroke between patients with and without malignancy. PATIENTS AND METHODS: This retrospective study involved patients with ischemic stroke admitted over 1 year to a Neurology Department. Demographic and clinical data, cardiovascular risk factors, and laboratory findings were recorded. The etiology was classified following the TOAST criteria. Participants were classified as stroke and cancer patients (diagnosis of malignancy before stroke or cancer diagnosed after admission) or as control stroke patients. RESULTS: Over 1 year, 291 stroke patients (268 ischemic) were admitted. Sixteen ischemic patients had a malignancy (5.9%), and in 12 patients, malignancy was the only cause found for the stroke; in 4 patients, malignancy was a coexisting condition. Coexistent stroke and cancer was significantly related to higher median levels of fibrinogen, D-dimers, and erythrocyte sedimentation rate (ESR). Logistic regression analysis showed that D-dimer and ESR levels were independently related to cancer, with odds ratios of 1.004 (95% CI 1.001-1.007, p=0.02) and 1.075 (95% CI 1.031-1.121, p=0.001), respectively. CONCLUSIONS: Almost 6% of ischemic stroke patients had a concomitant malignancy. In most of them, cancer was the only cause of stroke found. Patients with stroke and malignancy exhibited raised D-dimer, fibrinogen, and ESR levels, suggesting the prothrombotic state was a principal mechanism underlying stroke. D-dimer and ESR assessments may be useful in identifying underlying disorders in ischemic stroke.

Índices numéricos Ebrí-metacarpofalángico y carpiano para el cálculo de la edad ósea y predicción de talla adulta / Metacarpophalangeal and carpal numeric indices to calculate bone age and predict adult size

Introducción: En este trabajo se presenta unos nuevos métodos numéricos a partir de los índices metacarpofalángico y carpiano para el cálculo de la edad ósea, así como poder predecir la talla adulta por medio de ecuaciones multiregresión. Material y métodos: La casuística longitudinal comprende a 160 niños zaragozanos sanos de ambos sexos, con edades comprendidas desde los 6meses hasta los 20años, estudiados anualmente, incluido el estudio radiológico. Para la labor estadística se ha utilizado el paquete estadístico «Statistix», así como el programa Excel. Resultados: Los nuevos índices se encuentran íntimamente correlacionados con la edad cronológica, dando por ello lugar a ecuaciones predictivas para el cálculo de la edad ósea de niños hasta 20 años, presentándose además ecuaciones propias hasta los cuatro, a efectos de poder afinar más el diagnóstico a estas cortas edades. Las edades óseas calculadas pueden ser llevadas tanto a tablas numéricas de desviación típica, como a una figura de equivalencias que nos dan el diagnóstico osificativo directamente. Las ecuaciones predictivas de talla adulta permiten una estimación fiable de la talla futura del niño a estudio. Estas estimaciones analizadas por el test de Student no han dado diferencias significativas respecto a la talla adulta que los niños de la casuística al final alcanzaron. Los resultados pueden obtenerse con una calculadora o a través de un programa informático a disposición gratuita del lector. Conclusiones: Son presentadas por vez primera, por métodos propios no foráneos, estándares de edad ósea, así como ecuaciones de predicción de talla adulta para el estudio de niños. Se invita al clínico a utilizar estos métodos metacarpofalángico y carpiano a fin de conseguir la experiencia necesaria para su idónea aplicación en población sana y con diversa patología(AU)

Introduction: This work presents new numerical methods from the meta-carpal-phalangeal and carpal indexes, for calculating bone age. In addition, these new methods enable the adult height to be predicted using multiple regression equations. Materials and Methods: The longitudinal case series studied included 160 healthy children from Zaragoza, of both genders, aged between 6months and 20 years, and studied annually, including the radiological study. For the statistical analysis the statistical package "Statistix", as well as the Excel program, was used. Results: The new indexes are closely co-related to the chronological age, thus leading to predictive equations for the calculation of the bone age of children up to 20 years of age. In addition, it presents particular equations for up to 4years of age, in order to optimise the diagnosis at these early ages. The resulting bones ages can be applied to numerical standard deviation tables, as well as to an equivalences chart, which directly gives us the ossification diagnosis. The predictive equations of adult height allow a reliable forecast of the future height of the studied child. These forecasts, analysed by the Student test did not show significant differences as regards the adult height that children of the case series finally achieved. The results can be obtained with a pocket calculator or through free software available for the reader. Conclusions: For the first time, and using a centre-developed and non-foreign methods, bones age standards and adult height predictive equations for the study of children, are presented. We invite the practitioner to use these meta-carpal-phalangeal and carpal methods in order to achieve the necessary experience to apply it to a healthy population and those with different disorders(AU)

[Metacarpophalangeal and carpal numeric indices to calculate bone age and predict adult size]. / Índices numéricos Ebrí-metacarpofalángico y carpiano para el cálculo de la edad ósea y predicción de talla adulta.

INTRODUCTION: This work presents new numerical methods from the meta-carpal-phalangeal and carpal indexes, for calculating bone age. In addition, these new methods enable the adult height to be predicted using multiple regression equations. MATERIALS AND METHODS: The longitudinal case series studied included 160 healthy children from Zaragoza, of both genders, aged between 6 months and 20 years, and studied annually, including the radiological study. For the statistical analysis the statistical package "Statistix", as well as the Excel program, was used. RESULTS: The new indexes are closely co-related to the chronological age, thus leading to predictive equations for the calculation of the bone age of children up to 20 years of age. In addition, it presents particular equations for up to 4 years of age, in order to optimise the diagnosis at these early ages. The resulting bones ages can be applied to numerical standard deviation tables, as well as to an equivalences chart, which directly gives us the ossification diagnosis. The predictive equations of adult height allow a reliable forecast of the future height of the studied child. These forecasts, analysed by the Student test did not show significant differences as regards the adult height that children of the case series finally achieved. The results can be obtained with a pocket calculator or through free software available for the reader. CONCLUSIONS: For the first time, and using a centre-developed and non-foreign methods, bones age standards and adult height predictive equations for the study of children, are presented. We invite the practitioner to use these meta-carpal-phalangeal and carpal methods in order to achieve the necessary experience to apply it to a healthy population and those with different disorders.

Improvement of dermal burn healing by combining sodium alginate/chitosan-based films and low level laser therapy.

This paper aimed to evaluate the improvement of burn wounds healing by sodium alginate/chitosan-based films and laser therapy. Natural polymers with different biological activities are widely used as film dressings to improve wound healing. Lasers arrays accelerate the healing repair of soft tissue injuries. Burn procedures were performed on the backs of 60 male rats assigned into six groups: untreated (CTR), dressed with cellulose films (CL), dressed with sodium alginate/chitosan-based films (SC), laser-irradiated undressed wounds (LT), laser-irradiated wounds with cellulose (CLLT) and sodium alginate/chitosan-based films (SCLT). Laser therapy was applied for 7 days. Animals of each group were euthanised 8 and 14 days after the burn procedures. The inflammatory reaction was significantly more intense in the CTR group than in the irradiated groups after 8 and 14 days. Laser therapy stimulated myofibroblastic differentiation in 8 days, with or without dressing films. Combined laser therapy and both dressings improved epithelisation, blood vessels formation and collagenization, promoted rapid replacement of type III for type I collagen and favored the better arrangement of the newly formed collagen fibres. The combination of laser therapy and sodium alginate/chitosan-based dressing improves burn healing, apparently by modulating the epithelisation, blood vessels formation and collagenization processes.

Steady-state level of bone morphogenetic protein-15 in goat ovaries and its influence on in vitro development and survival of preantral follicles.

This study investigates steady-state level of bone morphogenetic protein-15 (BMP-15) mRNA in caprine follicles, and the effects of BMP-15 on in vitro development of preantral follicles. Ovarian fragments were cultured for one or seven days in Minimal Essential Medium (MEM(+)) with BMP-15 (0, 1, 10, 50, 100 or 200 ng/mL), and further analyzed by histology, transmission electron and fluorescent microscopy. BMP-15 mRNA in secondary follicles was higher than in primordial and primary follicles. After seven days, 10, 50 or 100 ng/mL of BMP-15 maintained the percentage of normal follicles similar to the control (non-cultured), and increased the oocyte and follicle diameters when compared to the control and MEM(+). BMP-15 at 100 ng/mL increased the secondary follicles and maintained their ultrastructural integrity. In conclusion, the BMP-15 mRNAs were detected in all follicular categories. BMP-15 (100 ng/mL) maintained the integrity and promoted the growth of caprine preantral follicles cultured for seven days.

Expression of follicle-stimulating hormone receptor (FSHR) in goat ovarian follicles and the impact of sequential culture medium on in vitro development of caprine preantral follicles.

This study evaluated the expression of FSH receptors (FSHR) in the different stages of goat follicle development and investigated whether the addition of increasing concentrations of FSH throughout the culture period influences the survival, growth and antral formation of in vitro-cultured caprine preantral follicles. The expression of FSHR was analysed before and after culturing follicles using real-time RT-PCR. For the culture, preantral follicles (≥150 µm) were isolated from ovarian fragments and cultured for 18 days in α-MEM+ alone or associated with recombinant FSH (rFSH: 100 or 1000 ng/ml), or in α-MEM+ supplemented with increasing concentrations of FSH throughout culture periods as follows: (a) sequential medium 1: FSH 100 ng/ml (from day 0 to 6), FSH 500 ng/ml (from day 6 to 12) and FSH 1000 ng/ml (from day 12 to 18); and (b) sequential medium 2: FSH 500 ng/ml (from day 0 to 9) and 1000 ng/ml (from day 9 to 18). Follicle development was evaluated on the basis of antral cavity formation, follicular and oocyte growth, and cumulus-oocyte complex health. The expression of FSHR in isolated caprine follicles increased from the preantral to antral phase. Regarding the culture, after 18 days, sequential medium 1 promoted follicular survival, antrum formation and a reduction in oocyte extrusion. Both sequential media promoted a higher rate of meiotic resumption compared with the other treatments. In conclusion, the addition of increased concentrations of FSH (sequential medium) has a significant impact on the in vitro development of caprine preantral follicles.

In vitro development of primordial follicles after long-term culture of goat ovarian tissue.

This study aims to investigate the effects of follicle stimulating hormone (FSH) and fibroblast growth factor-2 (FGF-2) on the survival and growth of caprine preantral follicles. Ovarian tissues were cultured for 1, 7, 14, 21 or 28 days in medium supplemented with FSH (FSH-2d or FSH-7d, i.e., with replacement of the culture medium every 2 or 7 days, respectively) or FSH+FGF-2 (replacement of the medium every 2 days). Non-cultured (control) and cultured ovarian fragments were processed for histological and ultrastructural analysis. After 28 days of culture, the media supplemented with FSH-2d was the most effective in maintaining the percentage of normal follicles and in promoting follicular growth. Furthermore, both treatments with FSH increased the percentage of the primary follicles. However, ultrastructural studies did not confirm follicular integrity from 14 days of culture onward. In conclusion, culturing tissue for up to 7 days in medium containing FSH alone or combined with FGF-2 maintains caprine preantral follicle integrity and promotes their growth in vitro.

Testosterone and cholesterol vasodilation of rat aorta involves L-type calcium channel inhibition.

Testosterone has rapid nongenomic vasodilator effects which could be involved in protective cardiovascular actions. Several authors suggested specific mechanisms to explain this effect, but this matter was not clarified yet. We studied the actions of testosterone and cholesterol on endothelium-denuded rat aorta and their effects on the L-type Ca(2+) current (I(Ca,L)) and potassium current (I(K)). Testosterone (1-100 µM) totally relaxed, in a rapid and concentration-dependent way, the aortic rings contracted by KCl or by (-)-Bay K8644 (BAY). Cholesterol also fully relaxed the contractions induced by KCl. None of the potassium channel antagonists tested (glibenclamide, tetraethylammonium and 4-aminopyridine) modified significantly the relaxant effect of testosterone. The antagonist of classic testosterone receptors, flutamide, did not modify the vasorelaxant effect of testosterone. Furthermore, testosterone and cholesterol inhibited either basal and BAY-stimulated I(Ca,L) in A7r5 cells and they have no effects on I(K). In summary, our results demonstrate that cholesterol and testosterone relax rat aorta by inhibiting LTCC. This effect of testosterone is not mediated by the classic hormone receptor or by potassium channel activation. These results suggest that the vasodilator mechanism of cholesterol and testosterone is the same.

Fibroblast growth factor-10 maintains the survival and promotes the growth of cultured goat preantral follicles.

The aim of the present study was to investigate the effects of fibroblast growth factor-10 (FGF-10) on the survival, activation (transition from primordial to primary follicles), and growth of goat preantral follicles cultured in vitro. Pieces of ovarian cortex were cultured for 1 and 7 d in the absence or presence of FGF-10 (0, 1, 10, 50, 100, and 200 ng/mL). Noncultured and cultured tissues were processed and analyzed by histology, transmission electron microscopy, and viability testing. Results showed that after 7 d, a greater percentage (79.9%) of morphologically normal follicles (containing an oocyte with regular shape and uniform cytoplasm, and organized layers of granulosa cells without a pyknotic nucleus) was observed when cultured with 50 ng/mL of FGF-10 when compared with other concentrations of FGF-10 (0 ng/mL, 67.3%; 1 ng/mL, 68.2%; 10 ng/mL, 63.3%; 100 ng/mL, 64.4%; 200 ng/mL, 52.7%). Ultrastructural analyses and viability testing using fluorescent markers confirmed the follicular integrity of FGF-10 (50 ng/mL)-treated fragments after 7 d of culture. After 7 d, all FGF-10 concentrations reduced the percentage of primordial follicles and increased the percentage of developing follicles. In the presence of 50 ng/mL of FGF-10, follicles increased in diameter after 7 d of culture when compared with other concentrations tested. In conclusion, this study demonstrates that FGF-10 maintains the morphological integrity of goat preantral follicles and stimulates the growth of activated follicles in culture. The culture conditions identified here contribute to the understanding of the factors involved in goat early follicular development.