RESUMO
The aim of this study was to assess the addition of 2% sodium caseinate in a commercial egg yolk-based medium in frozen ovine semen. Eight Dorper males were used for the study. The ejaculate was divided into two portions and frozen without (G1) or with the addition of 2% sodium caseinate (G2). Kinetic parameters were evaluated using CASA (computer-assisted sperm analysis), and membrane and acrosome integrity as well as oxidative stress were assessed using flow cytometry. After thawing, a thermoresistance test was conducted at time points T0 and T90. For the fertility test, 100 ewes were inseminated with semen from two rams selected based on in vitro parameters, one with good post-thaw quality (+70% total motility) and the other with low post-thaw quality (-55% total motility). For the fertility test, the females were divided into 4 groups for insemination: low-quality ram without caseinate (GBS = 25) and with caseinate (GBC = 25), and high-quality ram without caseinate (GAS = 25) and with caseinate (GAC = 25). Regarding the results of sperm kinetics, there was a statistically significant difference in the parameters of average path velocity (VAP) and curvilinear velocity (VCL) between the group frozen with BotuBov and the group with added caseinate. At time point T90, straight-line velocity maintained a trend (p < .06), with BotuBov® (BB group) being superior to caseinate this time, and in the linearity parameter, caseinate was superior to BotuBov®. Flow cytometry analysis showed no difference between any of the evaluated tests. In the fertility test, there was no statistically significant difference in the pregnancy rate between the BotuBOV® group (23%, 11/48) and the sodium caseinate group (BC group) (33%, 17/52), and no differences were observed in the male versus diluent interaction (p = .70). In conclusion, sodium caseinate supplementation did not influence sperm kinetic parameters and the fertility of sheep.
Assuntos
Caseínas , Criopreservação , Inseminação Artificial , Análise do Sêmen , Preservação do Sêmen , Motilidade dos Espermatozoides , Animais , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Masculino , Feminino , Criopreservação/veterinária , Criopreservação/métodos , Inseminação Artificial/veterinária , Caseínas/farmacologia , Análise do Sêmen/veterinária , Gravidez , Motilidade dos Espermatozoides/efeitos dos fármacos , Espermatozoides/efeitos dos fármacos , Espermatozoides/fisiologia , Crioprotetores/farmacologia , Sêmen/efeitos dos fármacos , Fertilidade/efeitos dos fármacos , Ovinos , Carneiro DomésticoRESUMO
This study aimed to assess the suitability of egg yolk (EY) supplementation to a tris-citric acid-based extender on cryosurvival of guinea pig (Cavia porcellus) epididymal spermatozoa. Two synthetic-based extenders, tris-citric acid-glucose plus 20% EY (TCG-EY) and tris-citric acid-fructose (TCF) both with 5% glycerol, were compared. Thirty-two epididymides were recovered from 16 adult guinea pig males by gonadectomy, and then the sperm samples were retrieved by retrograde flushing using TCG-EY and TCF extenders for left or right epididymis, respectively. TCG-EY and TCF sperm samples were frozen in static liquid nitrogen vapors through a two-step cooling procedure. Before freezing, the percentage of progressive sperm motility and sperm with intact plasma and acrosome membranes from TCG-EY sperm samples were higher (p < 0.05) than those diluted with TCF. Post-thaw sperm kinematic variables and membrane integrity were drastically reduced (p < 0.001) compared with prefreezing samples, regardless of extender type. The post-thaw plasmatic and acrosome membrane integrity from TCG-EY sperm samples was higher (p < 0.05) than those from TCF samples. Except for the length, the morphometric head dimensions of sperm diluted with TCG-EY or TCF did not vary (p > 0.05) after the freezing-thawing process compared with the prefreezing samples. In conclusion, despite greater cell cryoinjury with both extenders, the EY supplementation exerted greater cell membrane protection before and after the freezing-thawing process. This research shows an in-depth analysis of guinea pig sperm cryopreservation; however, more studies are recommended.
Assuntos
Gema de Ovo , Preservação do Sêmen , Masculino , Cobaias , Animais , Suínos , Epididimo , Ácido Cítrico/farmacologia , Motilidade dos Espermatozoides , Sêmen , Espermatozoides , Criopreservação/métodos , Preservação do Sêmen/métodos , Crioprotetores/farmacologiaRESUMO
OBJECTIVE: To evaluate the correlation between yolk sac diameter at 6 to 12 weeks of gestation measured via transvaginal ultrasound and adverse pregnancy outcomes. METHODS: This prospective cohort study was conducted at the Ain Shams University Maternity Hospital from July 1, 2019 to January 30, 2020. It included 120 pregnant women attending the outpatient clinic at 6 to 12 weeks of gestation. Transvaginal ultrasound was performed to measure inner yolk sac diameter. Normal diameter was considered to be 2-5 mm. Cases were followed up in routine antenatal care until the 16th week of gestation. RESULTS: Significant associations were found between maternal age and yolk sac diameter; yolk sac diameter and early miscarriage; a high percentage of cases of positive fetal life occurred when a normal yolk sac diameter (2-5 mm) was present (p<0.001); in yolk sac diameters <2mm positive fetal life was 0.0% and negative fetal life was 42.9%; in yolk sac diameters of 2-5mm positive fetal life was 81.1% and the negative fetal life was 7.1%; and in yolk sac diameters >5mm positive fetal life was 18.9% and negative fetal life was 50.0% (p<0.001), x2 60.094; and the best cutoff value for yolk sac diameter was >0.56, with a sensitivity of 78.6%, a specificity of 84.3%. CONCLUSIONS: We found a highly significant correlation between yolk sac diameter and early pregnancy loss.
Assuntos
Aborto Espontâneo , Resultado da Gravidez , Feminino , Gravidez , Humanos , Resultado da Gravidez/epidemiologia , Saco Vitelino/diagnóstico por imagem , Estudos Prospectivos , Ultrassonografia Pré-Natal , UltrassonografiaRESUMO
BACKGROUND: Hystricomorpha rodents display a similar placentation model to humans. The present study was carried out considering the scarcity of information concerning the placental development in agouti. OBJECTIVE: Describe the microscopy of the placenta, subplacenta and yolk sac of agoutis in early pregnancy and report on the inversion of the yolk sac. METHODS: Fifteen females between the 14th-32nd day of gestation were used following euthanasia. Gestational buttons were collected, fixed, processed, stained to optical microscopy or immunohistochemistry. RESULTS: Chorioallantoic placenta (CP) ranged from conical to a half-sphere, as follows: from the 14th to 17th day, the CP displays an inverted "V" shape, predominantly formed by cytotrophoblasts; from 20 to 22 days, formed almost entirely by cytotrophoblasts; at 28 days, a half sphere, with distinct lobes and interlobular area, numerous maternal gaps delimited by syncytiotrophoblasts and trophoblast giant cells; at 32 days, globose and undergoing the maturation process. Subplacenta, located between decidua and CP, initially presents septa consisting of simple columnar epithelium and after 17 days, comprising stratified epithelium. Visceral yolk sac (VYS) is attached to two CP projections between 14 and 17 days, formed by a simple cubic epithelium and inverted. Between 20 and 22 days, the epithelium displays apical villous projections with cytoplasmic vacuoles and a vascularized mesoderm. After the 24th day, the VYS near the placenta is pleated, very vascularized and villous, with decreased villi sizes further away from the placenta. CONCLUSION: The agouti CP displays similar characteristics to other hystricomorpha, including placenta lobulation, a subplacenta and an inverted vitelline placenta.
Assuntos
Dasyproctidae , Placentação , Gravidez , Feminino , Animais , Humanos , Placenta , Roedores , Saco VitelinoRESUMO
The use of assisted reproductive techniques, such as chilled semen, contributes to the maintenance and genetic improvement of canine breeding. The INRA-96 extender is a commercially available, chemically defined medium that was initially developed for the preservation of equine semen and exhibits preservation potential in the canine species. This research aims to evaluate the INRA-96 extender as an alternative for the short-term preservation of canine semen in terms of sperm quality parameters such as motility and kinetic parameters, integrity and functionality of the plasma membrane in fresh and chilled-rewarmed samples, as well as the sperm-binding ability using the perivitelline membrane of the chicken egg as an indicator of the fertilizing capacity of the preserved semen. A total of 18 ejaculates from 9 French bulldogs (two ejaculates per dog) were collected and divided into two aliquots that were diluted in Tris-egg yolk 20% (control) or INRA-96 to a final concentration of 100 × 106 sperm/mL. Samples were refrigerated in a biological incubator at 5°C and evaluated at 0, 24 and 48 h time points. Comparing the two treatments after 48 h of refrigeration, both extenders showed similar values (p < .5) for the majority of kinetic parameters, with the INRA-96 group promoting a total motility of 88.1 ± 2.9%. In addition, the morphology, integrity and functionality of the plasma membrane were preserved above 70% in this group. Dilution with INRA-96 also provided a significantly higher amount of sperm bound (256.2 ± 21.1) to the perivitelline membrane of the egg yolk compared to the sperm-binding rates (p < .05) achieved at the use of Tris-egg yolk (215.2 ± 21 bound spermatozoa) at 48 h. Our study proved similar functional properties of dog sperm cells treated with INRA-96 in comparison to commonly used home-made Tris-based extender during short-time storage.
Assuntos
Preservação do Sêmen , Sêmen , Animais , Cães , Masculino , Cavalos , Gema de Ovo/química , Benchmarking , Motilidade dos Espermatozoides , Preservação do Sêmen/veterinária , Preservação do Sêmen/métodos , Espermatozoides , Criopreservação/veterinária , Criopreservação/métodos , Crioprotetores/farmacologiaRESUMO
Temperature sex determination (TSD) in reptiles has been studied to elucidate the mechanisms by which temperature is transformed into a biological signal that determines the sex of the embryo. Temperature is thought to trigger signals that alter gene expression and hormone metabolism, which will determine the development of female or male gonads. In this review, we focus on collecting and discussing important and recent information on the role of maternal steroid hormones in sex determination in oviparous reptiles such as crocodiles, turtles, and lizards that possess TSD. In particular, we focus on maternal androgens and estrogens deposited in the egg yolk and their metabolites that could also influence the sex of offspring. Finally, we suggest guidelines for future research to help clarify the link between maternal steroid hormones and offspring sex.
Assuntos
Lagartos , Tartarugas , Animais , Masculino , Feminino , Estrogênios , Androgênios , Temperatura , Processos de Determinação Sexual , Tartarugas/genética , Esteroides , Diferenciação SexualRESUMO
Se presenta el caso clínico de una adolescente de 15 años de edad, quien fue asistida en el Hospital Provincial Pediátrico Universitario José Luis Miranda de Villa Clara, remitida desde su área de salud, por presentar dolor pélvico intenso desde hacía 3 días, náuseas y fiebre de 38,5 °C. Luego de realizados el examen clínico y los estudios complementarios pertinentes, se decidió practicar la resección completa del tumor. Durante el procedimiento se tomó muestra para estudio histológico que confirmó la existencia de un tumor del seno endodérmico ovárico, por lo cual fue reintervenida para extirpar el ovario contralateral y el epiplón infiltrados. Posteriormente se indicó poliquimioterapia según el protocolo y la evolución postratamiento fue satisfactoria.
The case report of a 15-years-old adolescent is presented, who was assisted at José Luis Miranda University Pediatric Provincial Hospital from Villa Clara, referred from her health area due to an intense pelvic pain for 3 days, nausea and fever of 38.5 °C. After carrying out the clinical exam and the pertinent laboratory tests, it was decided to practice the complete tumor resection. During the procedure a sample for histologic study was taken that confirmed the existence of an ovarian yolk sac tumor, reason why she was operated again to extirpate the contralateral ovary and the infiltrated omentum. Later on polychemotherapy was indicated according to the protocol and the post-treatment clinical course was satisfactory.
Assuntos
Neoplasias OvarianasRESUMO
Hydroxychloroquine sulfate (HCQ) and chloroquine diphosphate (CQ) have been used at increased rates to treat COVID-19 but can constitute a potential environmental risk. The objective was to evaluate the toxicity of sublethal concentrations of HCQ and CQ in zebrafish embryos/larvae. The 50% lethal concentrations (LC50) of HCQ and CQ at 96 h post-fertilization (hpf) were calculated by testing various concentrations on 2,160 embryos. The LC50 obtained were 560 and 800 µM for HCQ and CQ, respectively. Next, the embryotoxicity assay was performed, where 1,200 embryos were subjected to sublethal concentrations of HCQ and CQ. The hatching and heart rates were recorded. After euthanasia, photomicrographs of all larvae were taken to measure the total length, pericardial and yolk sac areas. The embryos exposed to sublethal concentrations of HCQ and CQ showed delayed hatching at 72 hpf, as well as an increase in the heart rate, larger pericardial and yolk sac areas, and body malformations at 96 hpf. The findings show that HCQ and CQ are toxic to fish in the early development phases. Understanding the mechanisms of toxicity will help extrapolate the effects of 4-aminoquinoline derivatives when they reach the aquatic environment in the context of the COVID-19 pandemic.
Assuntos
COVID-19 , Hidroxicloroquina , Animais , Humanos , Hidroxicloroquina/farmacologia , Peixe-Zebra , Pandemias , Tratamento Farmacológico da COVID-19 , LarvaRESUMO
Lipid droplets (LDs) are intracellular organelles found in most cell types from adipocytes to cancer cells. Although recent investigations have implicated LDs in numerous diseases, the current available methods to monitor them in vertebrate models rely on static imaging using fluorescent dyes, limiting the investigation of their rapid in vivo dynamics. Here, we report a fluorophore chemistry approach to enable in vivo LD dynamic monitoring using a Nernstian partitioning mechanism. Interestingly, the effect of atorvastatin and osmotic treatments toward LDs revealed an unprecedented dynamic enhancement. Then, using a designed molecular probe with an optimized response to hydration and LD dynamics applied to Zebrafish developing pericardial and yolk-sac edema, which represents a tractable model of a human cardiovascular disease, we also provide a unique dual method to detect disease evolution and recovery.
Assuntos
Corantes Fluorescentes , Gotículas Lipídicas , Animais , Humanos , Gotículas Lipídicas/química , Gotículas Lipídicas/metabolismo , Corantes Fluorescentes/química , Peixe-Zebra , Permeabilidade , Edema/metabolismoRESUMO
The aim of this study was to determine the influence of dietary phytase in breeder hens on yolk nutrients, hatchability, chick quality, and growth rate of their progeny, and their subsequent performance to 42 d post-hatch when fed diets with the same phytase concentrations. Breeder hens (n = 216) were divided into 3 groups receiving nutrient-adequate diets with reduced calcium (Ca) and phosphorus (P) (by 0.16% and 0.15%, respectively), supplemented with either 500, 1500, or 4500 FTU/kg phytase from 27 to 50 weeks of age. Eggs were collected at 38 weeks of age and incubated. On the day of hatch, the chick quality and hatchability were determined, and 18 chicks/group were euthanized for yolk sac collection and the determination of inositol and glycerol concentrations. The remaining chicks were divided into three groups, receiving different diets with reduced Ca and P (by 0.16% and 0.15%, respectively), supplemented with 0, 500, or 1500 FTU/kg phytase to 42 d post-hatch. Increasing the phytase concentration in the breeder hen diet linearly (p < 0.05) increased the number of early embryo deaths and decreased the number of late deaths and pips. The inositol concentration in the yolk sac at day of hatch increased (quadratic; p < 0.05) as the phytase dose increased in the breeder hen diet. The breeder hen diet (p < 0.05) influenced the body weight (BW), feed intake (FI), and feed conversion ratio (FCR) up to 21 days of age. The supplementation of breeder hen diets with 1500 FTU/kg phytase increased the concentration of sodium (Na), magnesium (Mg), potassium (K), manganese (Mn), and zinc (Zn) in the yolk sac. The inclusion of phytase doses up to 4500 FTU/kg appeared to influence embryo mortality, chick feed intake, and BW gain to 21 days and the FCR throughout the entire production phase.