Role of endothelin 1 in follicular and luteal function in the sheep

Three experiments were carried out in order to examine the role of endothelin in follicular steroid secretion in vitro and in spontaneous and PGF2alpha-induced luteolysis in sheep.;In experiment 1, it was hypothesized that endothelin 1 (END1) acting through its receptors ENDRA and/or ENDRB, on granulosal cells of ovine preovulatory follicles would inhibit steroid production, and therefore, prevent the premature luteinization of granulosal cells of those follicles. Data from estrogen-inactive 20 follicles collected from 13 ewes were analyzed. Accumulation of E 2 and P4 by cultured granulosal cells was not affected by either LH or END1 or the END receptor blockers. Granulosal cells from follicles with the greatest concentration of E2 in FF produced on the average significantly more E2 and P4 than follicles with lower concentrations of E2 in FF. In conclusion, END1 did not affect steroid production by granulosal cells from preovulatory follicles in sheep, and therefore the working hypothesis was not supported.;In the second experiment, the ENDRA antagonist BQ-610 was delivered into the CL via an osmotic minipump to examine the expression of genes related with P4 production and structural luteolysis at 6 and 24 h after exogenous PGF2alpha. It was hypothesized that sustained blockade of ENDRA would prevent upregulation of genes stimulated by PGF2alpha during structural luteolysis. Data for luteal weight, and concentrations of serum and luteal P4, mostly at 24 h after PGF2alpha, indicated that in 9 of 10 treated ewes, BQ-610 did not prevent luteolysis, and the pattern of gene expression confirmed this finding. Exogenous PGF2alpha downregulated expression of StAR, 3betaHSD, VEGF, TIMP-1 and eNOS (P ≤ 0.02), whereas it upregulated 20alphaHSD, MMP-14 Bcl-2 (P ≤ 0.03). Expression of Bcl-2 and Bax were reduced and Fas-R was increased from 6 to 24 h after PGF2alpha. In BQ-610-treated ewes, mRNA expression was upregulated more for 20alphaHSD, and caspases 3 and 8 than in vehicle-treated and control ewes. In conclusion, PGF2alpha induced luteolysis in both vehicle-treated and BQ-610-treated ewes.;In the third experiment, it was hypothesized that chronic delivery of BQ-610 into the CL would prevent luteal regression induced by endogenous PGF 2alpha, thereby increasing length of the estrous cycle and maintaining functional and structural characteristics of the CL. Ewes were assigned receive an osmotic mini-pump containing 2 mg of BQ-610 (n = 12) or vehicle (n = 9), implanted surgically on d 9 of the estrous cycle. Corpora lutea were collected 12 h after onset of estrus, or on the afternoon of d 21 in ewes that had not returned to estrus, and from untreated ewes on d 10 to 12 of the estrous cycle (mid-phase CL). Three of 12 BQ-610-treated ewes did not show estrus before d 21 compared to 0 of 9 vehicle-treated ewes (P = 0.33); CL of the remaining nine ewes treated with BQ-610 were excluded from further analysis. In conclusion, chronic infusion of BQ-610 blocked luteolysis and lengthened the estrous cycle in 3 of 12 ewes. Furthermore, functional features of CL of those 3 ewes were similar to mid-phase CL. Overall, these results supported a role for END1 in luteal regression in sheep. (Abstract shortened by UMI.).;Keywords. luteolysis, progesterone, END1, endothelin receptor type A, BQ-610, gene expression, sheep

Características morfológicas e histológicas de los ovarios de dos grupos genéticos de cuyes (Cavia porcellus) de América del Sur

This study aimed to describe morphological and histological characteristics of ovaries of two genetic groups of guinea pigs from Latin America. Ovaries from 20 improved and 20 native guinea pigs were collected after slaughtering. One ovary from each animal was weighed, measured, counted for visible follicles on the ovarian surface, and used for subsequent oocyte collection by the slicing method. Contralateral ovaries were used to prepare histological sections and quantify follicles. Body and ovary weight and ovary length were significantly greater in improved than in native guinea pigs (p<0.01). Ovarian weight was greater in diestrus than in proestrus (p=0.0632) only in improved animals. The number of primordial, primary, secondary, and total follicles did not differ between genetic groups. The number of antral follicles was significantly greater in improved than in native guinea pigs. The thickness of zona pellucida and oocyte diameter did not differ between groups. The thickness of the zona pellucida was significantly greater in oocytes of category A than B and C in both groups of guinea pigs. In conclusion, ovaries from improved guinea pigs were heavier and longer than those from native animals. The number of antral follicles was greater in improved than native guinea pigs. Zona pellucida thickness and oocyte diameter were similar in both groups of guinea pigs.El estudio tuvo como objetivo describir las características morfológicas e histológicas de los ovarios de dos grupos genéticos de cuyes de América Latina. Se recolectaron ovarios de 20 cuyes mejorados y 20 nativos después del sacrificio. Se pesó y midió un ovario de cada animal, se contaron los folículos visibles en la superficie ovárica y se colectaron los ovocitos mediante cortes seriados de la corteza ovárica. Los ovarios contralaterales se utilizaron para preparar cortes histológicos y cuantificar los folículos. El peso corporal y la longitud y peso de los ovarios fueron significativamente mayores en los cuyes mejorados que en los nativos (p<0.01). El peso de los ovarios fue mayor en diestro que en proestro (p=0.0632) en los animales mejorados. El número de folículos primordiales, primarios, secundarios y totales no difirió entre los grupos genéticos. El número de folículos antrales fue significativamente mayor en los cuyes mejorados que en los nativos. El grosor de la zona pelúcida y el diámetro de los ovocitos no difirieron entre los grupos. El grosor de la zona pelúcida fue significativamente mayor en los ovocitos de la categoría A que en los de la B y C en ambos grupos de cuyes. En conclusión, los ovarios de los cuyes mejorados fueron más pesados y largos que los de los animales nativos. El número de folículos antrales fue mayor en los cutes mejorados que en los nativos. El grosor de la zona pelúcida y el diámetro de los ovocitos fueron similares en ambos grupos de cuyes

Efecto de la adición de melatonina en el medio de maduración y/o vitrificación de ovocitos sobre la producción in vitro de embriones bovinos

The effect of the addition of melatonin (Mt) on the maturation and / or vitrification of bovine oocytes on cleavage and subsequent embryonic development was evaluated. Oocyte-cumulus cell complexes (COCs) were obtained from creole cows by ovum pick up (OPU) and by follicular aspiration from abattoir ovaries (AO). From the pool of oocytes obtained from both sources, those with homogenous cytoplasm and three or more compact layers of cluster cells were selected. The selected COCs were randomly assigned to five treatments: T1, matured with Mt and vitrified without Mt; T2, matured and vitrified with Mt; T3, matured without Mt and vitrified with Mt; T4 (control) matured and vitrified without Mt; T5, matured without Mt and not vitrified. The concentration of Mt in the maturation and vitrification media was 0.01 μM (10-9 M). The oocytes were matured, vitrified, fertilized and the presumed zygotes cultured until day 7 post-fertilization in vitro. The data were analysed by logistic regression. Regardless of the origin of the oocytes, cleavage rate (C) and embryo production (EP) was similar between treatments. The C in the OPU oocytes was greater in T4, and the EP was similar between treatments. In AO, the results did not vary between treatment in C and EP. In conclusion, Mt reduced the C in oocytes collected by OPU, while it did not affect EP. In the AO oocyte source, the addition of Mt did not affect the C nor the EP.Se evaluó el efecto de la adición de melatonina (Mt) en el medio de maduración y/o de vitrificación de ovocitos bovinos sobre clivaje y posterior desarrollo embrionario. Los complejos ovocito-células del cúmulo (COCs) fueron obtenidos de vacas criollas mediante la técnica de aspiración transvaginal guiada por ultrasonografía (OPU) y de ovarios de matadero (OM). Del pool de ovocitos obtenidos de ambas fuentes se seleccionaron los que tenían citoplasma homogéneo y tres o más capas compactas de células del cúmulo. Los COCs seleccionados fueron asignados aleatoriamente a cinco tratamientos: T1, madurados con Mt y vitrificados sin Mt; T2, madurados y vitrificados con Mt; T3, madurados sin Mt y vitrificados con Mt; T4 (control) madurados y vitrificados sin Mt; T5, madurados sin Mt y no vitrificados. La concentración de Mt en los medios de maduración y vitrificación fue de 0.01 μM (10-9 M). Los ovocitos fueron madurados, vitrificados, fecundados y los presuntos cigotos cultivados hasta el día 7 pos-fecundación in vitro. Los datos fueron analizados por regresión logística. Independientemente del origen de los ovocitos, el porcentaje de clivaje (PC) y de producción de embriones (PIV) fue similar entre tratamientos. El PC en los ovocitos de OPU fue mayor en T4, y la PIV fue similar entre tratamientos. En los de OM, los resultados no variaron entre tratamiento en PC y PIV. En conclusión, la Mt redujo el PC en ovocitos colectados por OPU, mientras que no afectó la PIV. En los colectados de OM la adición de Mt no afectó el PC ni la PI

Influencia del tamaño, la variedad y la proporción de sexos en la producción de huevos de tilapia (Oreochromis spp) en un sistema tropical intensivo al aire libre

The effect of the variety, size of the females and sex ratio in the production of eggs of Chitralada (CH: Oreochromis niloticus) and Taiwanese red (TR: O. mossambicus x O. niloticus) in outdoor concrete tanks were evaluated in a fish production unit located in the Andean foothills of the state of Trujillo, Venezuela. A 2x2x3 factorial arrangement design was used: two varieties (CH and TR), two sizes (large and small) and three female male proportions (1:1, 2:1 and 3:1). CH females produced greater volume and mass of eggs, spawned 2.4-fold more and were more fertile than TR (p<0.01). Large tilapia had 1.6-fold fewer spawning (p<0.05) and produced fewer eggs per 100 females than small ones (p<0.01). In general, the F:M ratio did not affect the reproductive variables. The small females of both varieties had a higher number of spawning than the large ones, and the large CH tilapia spawned 3.9 times more than the TR of the same size (p<0.01), while in the small ones this difference was only 1.7 times (p<0.05). The large CH tilapias were reproductively more efficient than the females of the same category of the TR variety. In conclusion, the CH tilapia and the smaller size and weight females were reproductively more efficient than those of the TR variety and the larger ones. Regardless of the variety, the reproductive activity was not affected by the proportions between females and males.Se evaluó el efecto de la variedad, tamaño de las hembras y relación hembra macho en la producción de huevos de tilapias Chitralada (CH: Oreochromis niloticus) y Taiwanesa roja (TR: O. mossambicus x O. niloticus), en tanques de concreto al aire libre, en una unidad de producción piscícola localizada en el Piedemonte Andino del estado Trujillo, Venezuela. Se utilizó un diseño en arreglo factorial 2x2x3: dos variedades (CH y TR), dos tamaños (grades y pequeñas) y tres proporciones hembra macho (1:1, 2:1 y 3:1). Las hembras CH produjeron mayor volumen y masa de huevos, desovaron 2.4 veces más y fueron más fecundas que las TR (p<0.01). Las tilapias grandes tuvieron 1.6 veces menos desoves (p<0.05) y produjeron menos huevos por cada 100 hembras que las pequeñas (p<0.01). En general, la relación H:M no afectó las variables reproductivas. Las hembras pequeñas de ambas variedades tuvieron un mayor número de desoves que las grandes, y las tilapias CH grandes desovaron 3.9 veces más que las TR del mismo tamaño (p<0.01), mientras que en las pequeñas esta diferencia fue de tan solo 1.7 veces (p<0.05). Las tilapias CH grandes fueron reproductivamente más eficientes que las hembras de la misma categoría de la variedad TR. En conclusión, las tilapias CH y las hembras de menor talla y peso fueron reproductivamente más eficientes que las de la variedad TR y las de mayor tamaño. Independientemente de la variedad, la actividad reproductiva no se vio afectada por las proporciones entre hembras y machos

Análisis morfométrico y funcional de ovocitos bovinos obtenidos de ovarios de matadero y por aspiración folicular transvaginal en vacas criollas del altiplano ecuatoriano

This study assessed the morphometric and functional characteristics of oocytes from 10 native cows to the Ecuadorian Andes, collected by transvaginal follicular aspiration (OPU) and oocytes collected post mortem (PM) from abattoir ovaries. For this purpose, 1157 cumulus-oocyte complexes (CCOs) of the OPU group (n=271) and PM group (n=886) were recovered in 10 repetitions per group by follicular aspiration at 90 mm Hg pressure and classified as A, B or C, according to the characteristics of the cytoplasm and cumulus cells. The activity of the enzyme glucose 6 phosphate dehydrogenase (G6PD) was determined as an indicator of follicular growth and maturation and the integrity of the plasma membrane by staining the CCOs with cresyl brilliant blue (BCB +) and trypan blue, respectively. The size (diameter and volume) of the oocytes was evaluated using CaptaVisio® software v. 5.1. The data were analysed by logistic regression and by the SAS general linear model, and the means were compared by the least square method. The results showed that the oocytes of the PM group had a larger diameter (126.0 ± 0.48 vs. 122.7 ± 0.79 µm; p<0.01) and integrity of the plasma membrane (86.0 vs. 76.8%; p<0.05) than the OPU group. However, quality A and B oocytes did not show significant differences between groups with respect to plasma membrane integrity and BCB + values. The results suggest that, although the fact that the size and viability were more affected in the oocytes aspirated by OPU, those of quality A and B from both sources showed similar values regarding the integrity of the plasma membrane and metabolic condition to continue. maturation.El estudio analizó las características morfométricas y funcionales de ovocitos de 10 vacas nativas de los Andes ecuatorianos, colectados por aspiración folicular transvaginal (OPU) y de ovocitos colectados post mortem (PM) de ovarios de matadero. Para este propósito, 1157 complejos cúmulo-ovocito (CCOs) del grupo OPU (n=271) y PM (n=886) fueron recuperados en 10 repeticiones por grupo por aspiración folicular a 90 mm Hg de presión y clasificados en calidad A, B o C, de acuerdo con las características del citoplasma y células del cúmulo. Se determinó la actividad de la enzima glucosa 6 fosfato deshidrogenasa (G6PD) como indicador de crecimiento folicular y maduración finalizado y la integridad de la membrana plasmática tiñendo los CCOs con azul brillante de cresilo (BCB+) y azul de tripán, respectivamente. Se evaluó el tamaño (diámetro y volumen) de los ovocitos usando un software CaptaVisionÒ (v. 5.1). Los datos se analizaron por regresión logística y por el modelo lineal general del SAS, y las medias se compararon por el método de los mínimos cuadrados. Los resultados mostraron que los ovocitos del grupo PM presentaron mayor diámetro (126.0±0.48 vs 122.7±0.79 µm; p<0.01) e integridad de la membrana plasmática (86.0 vs 76.8%; p<0.05) que el grupo OPU. Sin embargo, los ovocitos de calidad A y B no registraron diferencias significativas entre grupos con respecto a la integridad de la membrana plasmática y valores de BCB+. Los resultados sugieren que, a pesar de que el tamaño y la viabilidad fue más afectada en los ovocitos aspirados por OPU, los de calidad A y B provenientes de ambas fuentes mostraron valores similares con respecto a la integridad de la membrana plasmática y condición metabólica para proseguir la maduración

Acupuncture and rehabilitation of the painful shoulder: study protocol of an ongoing multicentre randomised controlled clinical trial [ISRCTN28687220]

BACKGROUND: Although the painful shoulder is one of the most common dysfunctions of the locomotor apparatus, and is frequently treated both at primary healthcare centres and by specialists, little evidence has been reported to support or refute the effectiveness of the treatments most commonly applied. According to the bibliography reviewed, physiotherapy, which is the most common action taken to alleviate this problem, has not yet been proven to be effective, because of the small size of sample groups and the lack of methodological rigor in the papers published on the subject. No reviews have been made to assess the effectiveness of acupuncture in treating this complaint, but in recent years controlled randomised studies have been made and these demonstrate an increasing use of acupuncture to treat pathologies of the soft tissues of the shoulder. In this study, we seek to evaluate the effectiveness of physiotherapy applied jointly with acupuncture, compared with physiotherapy applied with a TENS-placebo, in the treatment of painful shoulder caused by subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). METHODS/DESIGN: Randomised controlled multicentre study with blind evaluation by an independent observer and blind, independent analysis. A study will be made of 465 patients referred to the rehabilitation services at participating healthcare centres, belonging to the regional public health systems of Andalusia and Murcia, these patients presenting symptoms of painful shoulder and a diagnosis of subacromial syndrome (rotator cuff tendinitis and subacromial bursitis). The patients will be randomised into two groups: 1) experimental (acupuncture + physiotherapy); 2) control (TENS-placebo + physiotherapy); the administration of rescue medication will also be allowed. The treatment period will have a duration of three weeks. The main result variable will be the change produced on Constant's Shoulder Function Assessment (SFA) Scale; as secondary variables, we will record the changes in diurnal pain intensity on a visual analogue scale (VAS), nocturnal pain intensity on the VAS, doses of non-steroid anti-inflammatory drugs (NSAIDs) taken during the study period, credibility scale for the treatment, degree of improvement perceived by the patient and degree of improvement perceived by the evaluator. A follow up examination will be made at 3, 6 and 12 months after the study period has ended. Two types of population will be considered for analysis: per protocol and per intention to treat. DISCUSSION: The discussion will take into account the limitations of the study, together with considerations such as the choice of a simple, safe method to treat this shoulder complaint, the choice of the control group, and the blinding of the patients, evaluators and those responsible for carrying out the final analysis

Reproductive failure, possible maternal infanticide, and cannibalism in wild moustached tamarins, Saguinus mystax

Maternal infanticide in wild non-human primates has only been reported twice. In this paper, we report a possible new case of infanticide and cannibalism within a series of four successive reproductive failures in wild moustached tamarins, Saguinus mystax. Necropsy and genetic analyses of the corpses enabled us to rule out any pathology, and to determine paternity. The mother was seen biting and then eating the head of its own infant during a period when another female was pregnant and gave birth just 1 month later. Before that, the perpetrator had given birth to twins three times successfully when four to five adult and subadult males were present in the group. Although we do not know for certain that the infant was alive when the mother started biting it, our field observations preceding the event suggest it probably was. The possible infanticide case and the two cases of births and early death of the infants occurred while only two to three adult males were present in the group. This could be the second case of maternal infanticide reported in the genus Saguinus and the similar circumstances suggest a common pattern. We discuss these events in the light of the different functional explanations of infanticide and conclude that parental manipulation was the most likely: the mother could have terminated the investment in offspring that had low chances of survival in a group with low availability of helpers

TRY plant trait database – enhanced coverage and open access

Plant traits - the morphological, anatomical, physiological, biochemical and phenological characteristics of plants - determine how plants respond to environmental factors, affect other trophic levels, and influence ecosystem properties and their benefits and detriments to people. Plant trait data thus represent the basis for a vast area of research spanning from evolutionary biology, community and functional ecology, to biodiversity conservation, ecosystem and landscape management, restoration, biogeography and earth system modelling. Since its foundation in 2007, the TRY database of plant traits has grown continuously. It now provides unprecedented data coverage under an open access data policy and is the main plant trait database used by the research community worldwide. Increasingly, the TRY database also supports new frontiers of trait‐based plant research, including the identification of data gaps and the subsequent mobilization or measurement of new data. To support this development, in this article we evaluate the extent of the trait data compiled in TRY and analyse emerging patterns of data coverage and representativeness. Best species coverage is achieved for categorical traits - almost complete coverage for ‘plant growth form’. However, most traits relevant for ecology and vegetation modelling are characterized by continuous intraspecific variation and trait–environmental relationships. These traits have to be measured on individual plants in their respective environment. Despite unprecedented data coverage, we observe a humbling lack of completeness and representativeness of these continuous traits in many aspects. We, therefore, conclude that reducing data gaps and biases in the TRY database remains a key challenge and requires a coordinated approach to data mobilization and trait measurements. This can only be achieved in collaboration with other initiatives