In vivo and in vitro maturation of rabbit oocyte affects gene expression, mitochondrial distribution and apoptosis

In vivo-matured cumulus–oocyte complexes are valuable models in which to assess potential biomarkers of rabbit oocyte quality that contribute to enhanced IVM systems. In the present study we compared some gene markers of oocytes and cumulus cells (CCs) from immature, in vivo-matured and IVM oocytes. Moreover, apoptosis in CCs, nuclear maturation, mitochondrial reallocation and the developmental potential of oocytes after IVF were assessed. In relation to cumulus expansion, gene expression of gap junction protein, alpha 1, 43 kDa (Gja1) and prostaglandin-endoperoxide synthase 2 (Ptgs2) was significantly lower in CCs after in vivo maturation than IVM. In addition, there were differences in gene expression after in vivo maturation versus IVM in both oocytes and CCs for genes related to cell cycle regulation and apoptosis (V-Akt murine thymoma viral oncogene homologue 1 (Akt1), tumour protein 53 (Tp53), caspase 3, apoptosis-related cysteine protease (Casp3)), oxidative response (superoxide dismutase 2, mitochondrial (Sod2)) and metabolism (glucose-6-phosphate dehydrogenase (G6pd), glyceraldehyde-3-phosphate dehydrogenase (Gapdh)). In vivo-matured CCs had a lower apoptosis rate than IVM and immature CCs. Meiotic progression, mitochondrial migration to the periphery and developmental competence were higher for in vivo-matured than IVM oocytes. In conclusion, differences in oocyte developmental capacity after IVM or in vivo maturation are accompanied by significant changes in transcript abundance in oocytes and their surrounding CCs, meiotic rate, mitochondrial distribution and apoptotic index. Some of the genes investigated, such as Gja1, could be potential biomarkers for oocyte developmental competence in the rabbit model, helping improve in vitro culture systems in these species

Estudio de diferentes protocolos de restricción alimentaria sobre el desarrollo fetoplacentario en la coneja

Two feed strategies (ad libitum throughout pregnancy and restriction during the first 20 days), combined with oestrus synchronization with eCG or not in nulliparous rabbit does, were studied

La suplementación de los piensos de las conejas con EPA y DHA mejora el perfil insaturado de los ácidos grasos de la leche y sus parámetros reproductivos

Se ha valorado la influencia del enriquecimiento de las dietas de conejas reproductoras con ácidos grasos poliinsaturados de cadena larga (AGPI) de origen animal (EPA y DHA) durante 2 ciclos sobre sus parámetros reproductivos y la composición de su leche. Un total de 124 conejas se alimentaron desde la recría hasta el segundo destete con dos dietas isofibrosas, isoenergéticas e isoproteicas formuladas con dos fuentes de grasa distintas. El grupo control (C;n=62) recibió un pienso con un 3% de grasa mezcla mientras que el del grupo experimental (P;n=62) contenía un 6% de un suplemento con un 50% de extracto etéreo concentrado en DHA y EPA a partir de aceite de salmón atlántico (Optomega-50, Optivite International Ltd., Barcelona, España)

Alpha-tocopherol affects gene expression patterns of rabbit cumulus complexes and reduces apoptosis rate during in vitro maturation

Oxidative stress compromises oocyte developmental competence during in vitro maturation (IVM). Antioxidants such as vitamin E may avoid this imbalance. The aim of this study was to investigate the effect of a-Tocopherol (α-TocOH) on the relative mRNA abundance of genes involved in cumulus expansion (GJA1, PTGS2), cell cycle and viability (AKT1), cell cycle regulation and apoptosis (Tp53, CASP3) and antioxidant response (SOD2, GPX1, CAT) in rabbit cumulus oocyte complexes (COCs) in vitro matured. The apoptosis index in cumulus cells (CCs) and the hydrogen peroxide (H2O2) released by the COCs in maturation media were also assessed. For these purposes, COCs from follicles ≥1mm were recovered, selected and in vitro-matured for 16h (38ºC, 5% CO2) in a medium containing TCM-199 (Sigma, Madrid, Spain) with 0.3% bovine serum albumin (Sigma, Madrid, Spain) and 10 ng/mL Epidermal Growth Factor (EGF) (Sigma, Madrid, Spain) supplemented with 0, 100, 200 or 400 μM α- TocOH (Sigma, Madrid, Spain), named as 0E, 100E, 200E and 400E groups, respectivel

La posición fetal intrauterina afecta al desarrollo de las estructuras feto-placentarias de la coneja

Un total de 129 fetos de conejas multíparas fueron estudiados según su posición intrauterina. Los más próximos al ovario presentaron mejores valores morfométricos que los más alejados a esta posición, asociándose asimismo un mayor peso placentario a un mayor peso fetal. Estas diferencias fueron mantenidas en la valoración de órganos fetales, como el cerebro, hígado y aparato digestivo, mostrándose un mayor desarrollo en los fetos adyacentes al ovario. A su vez se observó una correlación positiva entre el peso placentario y el peso de estos órganos. Las diferencias de peso dentro de la misma camada podrían estar asociadas a un mayor desarrollo placentario y por consiguiente mayor disponibilidad de nutrientes

Design and experimental validation of a magnetic device for stem cell culture

Cell culture of bone and tendon tissues requires mechanical stimulation of the cells in order to mimic their physiological state. In the present work, a device has been conceived and developed to generate a controlled magnetic field with a homogeneous gradient in the working space. The design requirement was to maximize the magnetic flux gradient, assuring a minimum magnetizing value in a 15 mm × 15 mm working area, which highly increases the normal operating range of this sort of devices. The objective is to use the machine for two types of biological tests: magnetic irradiation of biological samples and force generation on paramagnetic particles embedded in scaffolds for cell culture. The device has been manufactured and experimentally validated by evaluating the force exerted on magnetic particles in a viscous fluid. Apart from the magnetic validation, the device has been tested for irradiating biological samples. In this case, viability of human dental pulp stem cells has been studied in vitro after electromagnetic field exposition using the designed device. After three days of irradiation treatment, cellular microtissues showed a 59% increase in the viable cell number. Irradiated cells did not show morphological differences when compared with control cells

The effects of sildenafil citrate on the fetoplacental development and hemodynamics in a rabbit model of intrauterina growth restriction

The present study evaluated the effectiveness of sildenafil citrate (SC) to improve placental and fetal growth in a diet-induced rabbit model of intrauterine growth restriction (IUGR). Pregnant rabbits were fed either ad libitum (Group C) or restricted to 50% of dietary requirements (Group R) or restricted and treated with SC (Group SC). The treatment with SC improved placental development by increasing vascularity and vessel hypertrophy in the decidua. The assessment of feto–placental haemodynamics showed higher resistance and pulsatility indices at the middle cerebral artery (MCA) in fetuses treated with SC when compared with Group R, which had increased systolic peak and time-averaged mean velocities at the MCA. Furthermore, fetuses in the SC group had significantly higher biparietal and thoracic diameters and longer crown–rump lengths than fetuses in Group R. Hence, the SC group had a reduced IUGR rate and a higher kit size at birth compared with Group R. In conclusion, SC may provide potential benefits in pregnancies with placental insufficiency and IUGR, partially counteracting the negative effects of food restriction on placental development and fetal growth. However, the present study also found evidence of a possible blood overflow in the brain that warrants further investigation

Point-of-care lung ultrasound assessment for risk stratification and therapy guiding in COVID-19 patients. A prospective non-interventional study.

Background Lung ultrasound is feasible for assessing lung injury caused by coronavirus disease 2019 (COVID-19). However, the prognostic meaning and time-line changes of lung injury assessed by lung ultrasound in COVID-19 hospitalised patients are unknown. Methods Prospective cohort study designed to analyse prognostic value of lung ultrasound in COVID-19 patients by using a quantitative scale (lung ultrasound Zaragoza (LUZ)-score) during the first 72 h after admission. The primary end-point was in-hospital death and/or admission to the intensive care unit. Total length of hospital stay, increase of oxygen flow and escalation of medical treatment during the first 72 h were secondary end-points. Results 130 patients were included in the final analysis; mean±sd age was 56.7±13.5 years. Median (interquartile range) time from the beginning of symptoms to admission was 6 (4–9) days. Lung injury assessed by LUZ-score did not differ during the first 72 h (21 (16–26) points at admission versus 20 (16–27) points at 72 h; p=0.183). In univariable logistic regression analysis, estimated arterial oxygen tension/inspiratory oxygen fraction ratio (PAFI) (hazard ratio 0.99, 95% CI 0.98–0.99; p=0.027) and LUZ-score >22 points (5.45, 1.42–20.90; p=0.013) were predictors for the primary end-point. Conclusions LUZ-score is an easy, simple and fast point-of-care ultrasound tool to identify patients with severe lung injury due to COVID-19, upon admission. Baseline score is predictive of severity along the whole period of hospitalisation. The score facilitates early implementation or intensification of treatment for COVID-19 infection. LUZ-score may be combined with clinical variables (as estimated by PAFI) to further refine risk stratification

Multiple approaches at admission based on lung ultrasound and biomarkers improves risk identification in COVID-19 patients

Background: Risk stratification of COVID-19 patients is fundamental to improving prognosis and selecting the right treatment. We hypothesized that a combination of lung ultrasound (LUZ-score), biomarkers (sST2), and clinical models (PANDEMYC score) could be useful to improve risk stratification. Methods: This was a prospective cohort study designed to analyze the prognostic value of lung ultrasound, sST2, and PANDEMYC score in COVID-19 patients. The primary endpoint was in-hospital death and/or admission to the intensive care unit. The total length of hospital stay, increase of oxygen flow, or escalated medical treatment during the first 72 h were secondary endpoints. Results: a total of 144 patients were included; the mean age was 57.5 ± 12.78 years. The median PANDEMYC score was 243 (52), the median LUZ-score was 21 (10), and the median sST2 was 53.1 ng/mL (30.9). Soluble ST2 showed the best predictive capacity for the primary endpoint (AUC = 0.764 (0.658–0.871); p = 0.001), towards the PANDEMYC score (AUC = 0.762 (0.655–0.870); p = 0.001) and LUZ-score (AUC = 0.749 (0.596–0.901); p = 0.002). Taken together, these three tools significantly improved the risk capacity (AUC = 0.840 (0.727–0.953); p = 0.001). Conclusions: The PANDEMYC score, lung ultrasound, and sST2 concentrations upon admission for COVID-19 are independent predictors of intra-hospital death and/or the need for admission to the ICU for mechanical ventilation. The combination of these predictive tools improves the predictive power compared to each one separately. The use of decision trees, based on multivariate models, could be useful in clinical practice. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/)