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ObjectiveThis study aimed to develop a novel method for encapsulating oocytes in sodium alginate hydrogel using microfluidics, then to vitrify these encapsulated oocytes in a single-step process with low concentrations of cryoprotectants. MethodsWe utilized a flow-focusing microfluidic chip to generate sodium alginate hydrogel microspheres. The influence of various parameters, including throat structure, cross-linking method, sodium alginate concentrations, and flow rate ratios on the stability diameter, and coefficient of variation of microspheres were examined. To further investigate the cold-resistance of these microspheres, we used cryomicroscopy to observe changes in volume and morphology of microspheres during cooling and warming processes. We used microfluidic chip to encapsulate oocytes in sodium alginate hydrogel microspheres, the empty rate of microspheres and loss rate of oocytes were determined. After releasing from microspheres and parthenogenetic activation with cytochalasin B and strontium chloride, the survival, cleavage and blastocyst rates were evaluated during in vitro maturation. Finally, oocytes encapsulated in sodium alginate microspheres were vitrified with low concentrations of cryoprotectants. We compared the survival and development capability of the oocytes with the Cryotop method. ResultsWhen the throat of the microfluidic chip measures 300 μm in length and 120 μm in width, microspheres can be uniformly formed at the throat of the chip. Sodium alginate generates microspheres with a wide size distribution when cross-linking outside the chip, while internal cross-linking within the chip results in more uniform microspheres. The stability of microsphere formation is significantly improved with the use of a three-channel internal cross-linking chip. At a flow rate of 2 μl/min and with 1% sodium alginate, the microfluidic chip can consistently and uniformly produce microspheres. Under flow rate ratios of 10, 15, and 20, the average microsphere diameters are 262.71 μm, 193.63 μm, and 156.63 μm, respectively. The sodium alginate hydrogel microspheres maintained their volume and structural integrity during the cooling and warming processes. Using a three-channel internal cross-linking microfluidic chip to encapsulate oocytes, at a flow rate ratio of 10, the empty rate is 32.28%, and the cell loss rate is 11.09%. After encapsulation and subsequent release, the oocyte survival rate (96.99%), cleavage rate (88.71%), and blastocyst formation rate (26.29%) showed no significant differences compared to the fresh group. After the microspheres were vitrified using a low concentration of cryoprotectant (10% DMSO+10% ehylene glycol (EG)+0.5 mol/L trehalose), the survival rate, cleavage rate, and blastocyst rate were 92.48%, 70.80%, and 20.42%, respectively. No significant difference was observed when compared to the Cryotop method using a higher concentration of cryoprotectant solution (15% DMSO+15% EG+0.5 mol/L trehalose). ConclusionWe designed and fabricated a microfluidic system with three-channel internal cross-linking chips used for oocyte vitrification preservation. The microfluidic system can generate oocytes-loaded sodium alginate hydrogel microspheres with uniform size, low empty rate, and good cold-resistance. The method successfully reduced the concentration of cryoprotectants in a single-step vitrification process, the developmental capability of oocytes during in vitro maturation were comparable with Cryotop method. Unlike the Cryotop method, the oocytes encapsulated in hydrogel does not come into contact with liquid nitrogen, eliminating the risk of cross-contamination. This study provides a novel approach to oocyte vitrification.
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O hormônio antimulleriano é secretado pelas células da granulosa dos folículos que estão em desenvolvimento no ovário. Por meio da sua dosagem, é possível avaliar a reserva ovariana. A mulher tem seu número máximo de oócitos no perío- do fetal, mas, conforme o tempo passa, existe uma queda do número de células germinativas. Desse modo, para mulheres que têm o desejo de engravidar, a dosa- gem de hormônios e a avaliação da reserva ovariana podem ajudar no processo. O objetivo do estudo foi encontrar evidências na literatura que comprovem que o hormônio antimulleriano é o melhor marcador da reserva ovariana. Para isso, foi realizada uma revisão integrativa, classificada como qualitativa; a busca de da- dos foi realizada no PubMed, utilizando a seguinte palavra-chave: "hormônio anti- mulleriano (HAM)". Foram encontrados oito artigos que abordavam diretamente o tema, e há evidências que corroboram a hipótese de que o hormônio antimulleria- no é um bom marcador da reserva ovariana, sendo necessários mais estudos para determinar a sua superioridade.
The anti-mullerian hormone is secreted by the granulosa cells of follicles that are developing in the ovary. Though its dosage is possible to evaluate the ovarian re- serve. Women have their maximum number of oocytes in the fetal period, but there is a decrease in the number of germinative cells as time goes by. Thus, women that desire to get pregnant can have hormones dosed and the ovarian reserve evalua- ted to help them with this process. The objective of this study was to find evidence in the literature that proves that the anti-mullerian hormone is the best marker of ovarian reserve. For this purpose, an integrative review was conducted, using the key word: "anti-mullerian hormone (AMH)". Eight articles were found on the subject and there is evidence that proves the hypothesis of the anti-mullerian hormone as a good marker, however more studies are needed to determine its superiority.
الموضوعات
Humans , Female , Pregnancy , Anti-Mullerian Hormone/chemistry , Ovarian Reserve/physiology , Oocytes , Cell Count/methods , Women's Health , Fertilityالملخص
Introducción La vitrificación de óvulos es un avance científico que ha cambiado la perspectiva reproductiva de la sociedad humana. Este procedimiento se ha ofrecido como alternativa a la postergación voluntaria del embarazo, confiriéndole a la mujer una nueva perspectiva en su autonomía reproductiva. El número de mujeres que consultan y luego optan por congelar ovocitos ha aumentado en forma casi exponencial en Chile y en todo el mundo. En nuestro país, hay poco conocimiento acerca de la motivación, experiencia y resultados de la criopreservación electiva de ovocitos en Chile. El objetivo fue conocer la motivación, experiencia y el deseo reproductivo futuro de este grupo de mujeres sometidas a esta técnica. Métodos Estudio descriptivo transversal, basado en un cuestionario enviado por correo electrónico en el que participaron mujeres que se habían sometido previamente a criopreservación electiva de ovocitos entre enero de 2011 y diciembre de 2019 en Clínica Alemana, Santiago de Chile. Resultados De 342 mujeres que habían completado un ciclo de criopreservación electiva de ovocitos, 193 aceptaron participar y de estas, 98 (51%) de las mujeres contestaron la encuesta en forma satisfactoria. Se establecieron criterios de exclusión a aquellas mujeres que se habían sometido a este procedimiento por indicación médica como la endometriosis, el cáncer y la baja reserva ovárica. El motivo más frecuente para realizarse el procedimiento fue la edad (44%). En relación al procedimiento; el 94% no se arrepiente de haberlo realizado y 74% de las mujeres cree que utilizará sus ovocitos en algún momento de su vida. Por último, desde que se realizaron la criopreservación de ovocitos a la fecha, el 11% de las mujeres encuestadas ha usado sus ovocitos vitrificados y 27% ha logrado embarazarse con estos. Conclusión Las mujeres que se someten a criopreservación electiva de ovocitos por razones sociales, son principalmente mujeres sin pareja que tiene como motivación principal su edad reproductiva y la gran mayoría de ellas no se arrepienten de haberlo realizado.
Introduction Oocyte vitrification is a scientific advance that has changed the reproductive perspective of human society. This procedure has been offered as an alternative to the voluntary postponement of pregnancy, giving women a new perspective on their reproductive autonomy. The number of women who consult and then choose to freeze oocytes has increased almost exponentially in Chile and throughout the world. There is little knowledge about the motivation, experience, and results of elective oocyte cryopreservation in Chile. The objective was to know the motivation, experience, and future reproductive desire of the women who underwent this technique. Methods Cross-sectional descriptive study based on a questionnaire sent by e-mail in which females who had previously undergone elective oocyte cryopreservation between January 2011 and December 2019 at Clínica Alemana, Santiago, Chile, participated. Results Of 342 women who had completed a cycle of elective oocyte cryopreservation, 193 agreed to participate, and of these, 98 (51%) answered the survey satisfactorily. Women who underwent this procedure for medical indication, including endometriosis, cancer, and low ovarian reserve, were excluded. The most frequent reason for the procedure was age (44%). Concerning the procedure: 94% do not regret having it done, and 74% of the women believe that they will use their oocytes at some point in their lives. Finally, from the time of oocyte cryopreservation to date, 11% of the surveyed women have used their vitrified oocytes, and 27% have become pregnant. Conclusions Women who undergo elective oocyte cryopreservation for social reasons are mainly women without a partner whose main motivation is their reproductive age. The vast majority do not regret doing so.
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OBJECTIVE@#To explore the effects of Zishen Yutai Pills (ZYPs) on the quality of oocytes and embryos, as well as pregnancy outcomes in patients with diminished ovarian reserve (DOR) receiving in vitro fertilization-embryo transfer (IVF-ET). The possible mechanisms, involving the regulation of bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9), were also investigated.@*METHODS@#A total of 120 patients with DOR who underwent their IVF-ET cycle were randomly allocated to 2 groups in a 1:1 ratio. The patients in the treatment group (60 cases) received ZYPs from the mid-luteal phase of the former menstrual cycle by using gonadotropin-releasing hormone (GnRH) antagonist protocol. The patients in the control group (60 cases) received the same protocol but without ZYPs. The primary outcomes were the number of oocytes retrieved and high-quality embryos. Secondary outcomes included other oocyte or embryo indices as well as pregnancy outcomes. Adverse events were assessed by comparison of the incidence of ectopic pregnancy, pregnancy complications, pregnancy loss, and preterm birth. Contents of BMP15 and GDF9 in the follicle fluids (FF) were also quantified with enzyme-linked immunosorbent assay.@*RESULTS@#Compared with the control group, the numbers of oocytes retrieved and high-quality embryos were significantly increased in the ZYPs group (both P<0.05). After treatment with ZYPs, a significant regulation of serum sex hormones was observed, including progesterone and estradiol. Both hormones were up-regulated compared with the control group (P=0.014 and 0.008), respectively. No significant differences were observed with regard to pregnancy outcomes including implantation rates, biochemical pregnancy rates, clinical pregnancy rates, live birth rates, and pregnancy loss rates (all P>0.05). The administration of ZYPs did not increase the incidence of adverse events. The expressions of BMP15 and GDF9 in the ZYPs group were significantly up-regulated compared with the control group (both P<0.05).@*CONCLUSIONS@#ZYPs exhibited beneficial effects in DOR patients undergoing IVF-ET, resulting in increments of oocytes and embryos, and up-regulation of BMP15 and GDF9 expressions in the FF. However, the effects of ZYPs on pregnancy outcomes should be assessed in clinical trials with larger sample sizes (Trial reqistration No. ChiCTR2100048441).
الموضوعات
Infant, Newborn , Pregnancy , Female , Humans , Fertilization in Vitro/methods , Ovarian Reserve , Prospective Studies , Premature Birth , Embryo Transfer/methods , Ovulation Induction/methods , Gonadotropin-Releasing Hormone/therapeutic useالملخص
ObjectiveTo observe the regulation of Qigongwan on the expression of proliferation and apoptosis-related factors programmed cell death 4 (PDCD4) and proliferating cell nuclear antigen (PCNA) in ovarian granulosa cells (GCs) in patients with polycystie ovarian syndrome (PCOS) infertility with phlegm-dampness syndrome, and to explore the effect of Qigongwan on the quality of oocytes and embryonic development potential. MethodSixty-six patients with PCOS with phlegm-dampness syndrome who underwent in vitro fertilization and embryo transfer (IVF-ET) were randomly selected and divided into an observation group (Qigongwan + western medicine) and a control group (western medicine), with 33 patients in each group. Antagonist regimen was used to promote ovulation in the two groups. The observation group was given Qigongwan one cycle before IVF based on the treatment of conventional western medicine, while the control group was not given Chinese medicine. The improvement of phlegm and dampness syndrome, the dosage and the number of days of using gonadotropins (Gn), the levels of luteinizing hormone (LH), estradiol (E2), and progesterone (P) on the day of human chorionic gonadotropin(HCG) injection, the 2PN fertilization rate, and the high-quality embryo rate of patients in the two groups were compared. Real-time polymerase chain reaction (Real-time PCR) and Western blot technology were used to detect the expression of PCNA and PDCD4 in GCs. ResultAs compared with groups before treatment, the score of phlegm-dampness syndrome in both groups was significantly lower (P<0.01). The score of phlegm and dampness syndrome in the observation group was significantly lower than that of the control group (P<0.01). As compared with the control group, the levels of LH, E2, and P in the observation group was higher, but only the difference in the level of E2 was statistically significant (P<0.01). The 2PN fertilization rate [82.25% (556/676) vs 69.92% (365/522), χ2=25.172, P<0.01] and high-quality embryo rate [44.19% (190/430) vs 34.23% (102/298), χ2=7.266, P<0.01] in the observation group were significantly higher than that of the control group (P<0.01). As compared with the control group, the mRNA and protein expression of PDCD4 in ovarian GCs was down-regulated in the observation group and that of PCNA was up-regulated (P<0.05). ConclusionBy down-regulating the expression of PDCD4 and up-regulating the expression of PCNA, Qigongwan may interfere with follicle development, adjust hormone levels, improve the symptomatic manifestations of patients with PCOS with phlegm-dampness syndrome, inhibit the apoptosis of GCs, and promote growth, thus improving the quality of oocytes and embryonic development potential.
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Objective @#To investigate the effects of 3 types of trihalomethanes ( THMs) , the by⁃products in treatment of drinking water disinfection , on the development and reproductive capacity of germ cells of Caenorhabditis elegans. @*Methods @#Tribromomethane ( TBM) , trichloromethane ( TCM) and triiodomethane ( TIM) , 3 types of THMs were selected and wild type Caenorhabditis elegans was used as a model to detect germ cell apoptosis , oocyte number and the final brood size after exposure to different concentrations of THMs.@*Results @#The results of germ cell apoptosis showed that the exposure to 50 μmol/L TCM and TIM caused elevated levels of germ cell apoptosis in a dose⁃dependent manner. Oocyte counting results showed that exposure to 50 μmol/L all three THMs significantly reduced the number of oocytes. The brood size results showed that only 50 μmol/L TIM exposure significantly de⁃ creased the brood size of nematodes.@*Conclusion @#THMs showed negative impacts on the reproductive system of Caenorhabditis elegans , and the toxicity is related to the halogenated elements of THMs.
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Aims: Reproductive parameters are important tools for the sustainable management of halieutic species in the water bodies of its exploitation. This study intends to characterize the sexual maturity scale of swimming crab Callinectes pallidus, perform testicular and ovarian structures in order to develop a specific maturity scale. Study Design: This study was carried out with macroscopic and microscopic observations of 394 specimens of crabs including 206 females and 188 males. Place and Duration of Study: The specimens used in this study were collected monthly from March to July 2018 from small scale fisheries of Lake Nokou� (Benin). Methodology: At the laboratory, each crab specimen is identified and its sex determined. Grids for describing each sexual maturity stage of the crab C. pallidus were developed based on the Zairon et al. (2015) scale. Histological sections of female and male gonads were analyzed in order to certify the different stages of sexual development noted from the macroscopic study. Results: Macroscopic analysis of external and internal anatomy and histological study of male and female gonads of C. pallidus allowed to establish a sexual maturity scale with 5 stages in females and 4 stages in males respectively. Macroscopic observations made with a binocular magnifying glass allowed to describe the characteristic features of the sexual development cycle of the species. The histological study confirmed the results of the macroscopic analysis. Significant difference was recorded between the mean oocytes diameters of the different oocyte development stages (p<0.05). Conclusion: The outcomes of this constitute an important database for the assessment of the reproductive parameters of the species in the water bodies of its exploitation. The stages IV and V are the mature steps in the female development whereas the stages III and IV are that of the male. It is recommended to allow C. pallidus reach the sexual maturity size (5,95 cm for female and 6,38 cm for male) before their exploitation to ensure the sustainable management of the species.
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Objective:To investigate the relationship between the level of anti Mullerian hormone (AMH) and antral follicle count (AFC) and oocytes number in patients with in vitro fertilization (IVF). Methods:372 patients with primary infertility who received IVF and embryo transfer (IVF-ET) treatment in the Affiliated Hospital of Guizhou Medical University from January 2020 to December 2020 were prospectively selected as the study subjects. According to the oocytes obtained, the patients were divided into low ovarian response group (LOR group, the number of oocytes obtained ≤5, n=37), normal group (NOR group, the number of oocytes obtained was 6-15, n=292) and high ovarian response group (HOR group, the number of oocytes obtained >15, n=43). The levels of AMH and AFC in the three groups were observed. The relationship between AMH, AFC with LOR and HOR was observed by multivariate logistic analysis. The sensitivity, specificity, optimal cut-off value and area under the curve (AUC) of AMH and AFC for predicting LOR and HOR were calculated by receiver operating characteristic (ROC) curve. Results:There were significant difference in AMH, AFC and oocyte number among the three groups (all P<0.05). The AMH, AFC and oocyte number in NOR group were higher than those in LOR group, and AMH, AFC and oocyte number in HOR group were higher than those in LOR and NOR group (all P<0.05). The results of binary multivariate analysis showed that AFC and AMH were protective factors of LOR (all P<0.05), while AFC and AMH were risk factors of HOR ( P<0.05). The sensitivity and specificity of AMH and AFC in predicting LOR were 83.8% and 75.7%, 65.7% and 84.2%, respectively. The sensitivity of AMH or AFC positive as the standard for predicting LOR was 94.6%, and the specificity of AMH and AFC was 91.6%. The sensitivity and specificity of positive AMH and AFC in predicting HOR were 69.8% and 74.4%, 69.6% and 83.6%, respectively. The sensitivity of AMH or AFC positive as the standard for predicting HOR was 93.0%, and the specificity of AMH and AFC positive as the standard for predicting HOR was 93.0%. Conclusions:AMH and AFC are significantly correlated with oocyte number in patients with primary infertility. Flexible application of AMH and AFC is beneficial to better predict the number of oocytes obtained, which provides a reference for clinical development of individualized ovarian stimulation program.
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The objective of this study was to quantify the number and frequency of monocyte (MnOF) and multi-oocyte (MtOF) follicles in ovaries of bitches subjected to ovary salpingohysterectomy (OSH). Right and left ovaries of 38 bitches were collected after OSH, prepared, and a histological analysis was carried out. The ovaries were subjected to surface and deep histological cuts; the follicles were classified, and the number of follicles and cumulus oophorus complexes (COC) per follicle were quantified for each histological cut. MnOF and MtOF were found in all ovaries, at different developmental stages; primary follicles were grouped in the ovarian cortex, and follicles at other follicular stages presented a random distribution. MtOF containing two, three, four, or more COC were found in the ovaries of bitches, with a decreasing frequency trend, according to the number of COC in the MtOF. The effect of the age, number of estrus, estrus interval, and number of progenies per delivery was not significant for the number and frequency of MtOF in the ovaries of the bitches, whereas the size, number of pregnancies, use and number of contraceptive applications had some effect on the number and frequency of MtOF in the ovaries of the bitches.(AU)
Objetivou-se, com este estudo, quantificar o número e a frequência de folículos monocitários (MOF) e polioocitários (POF) provenientes de ovários de cadelas submetidas à ovariossalpingo-histerectomia (OSH). Para tanto, coletaram-se os ovários (direito e esquerdo) de 38 cadelas após OSH, com posterior preparação e análise histológica. Cada ovário foi submetido a dois cortes histológicos (superficial e profundo) onde se quantificou o número e a classificação dos folículos, bem como o número de complexos cumulus oophorus (COCs) por folículo em cada corte histológico. Observaram-se MOF e POF em todos os ovários estudados, em diferentes estádios de desenvolvimento, sendo os folículos primários agrupados no córtex ovariano, frente a uma distribuição aleatória dos outros estádios foliculares. FOPs contendo dois, três, quatro ou mais COCs foram observados nos ovários de todas as fêmeas estudadas, e sua frequência tendeu a diminuir de acordo com o número de COC presente no POF. Não se observou influência da idade, do número e do intervalo de estros, assim como do número de filhotes por gestação sobre o número/frequência de FOP nos ovários das cadelas estudadas, enquanto o porte, o número de gestações, o uso e o número de contraceptivo apresentaram algum grau de influência sobre o número/frequência de FOP nos ovários das cadelas estudadas.(AU)
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Animals , Female , Cats , Oocytes/classification , Cumulus Cells/classification , Ovarian Follicle , Periodicity , Ovariectomy/veterinary , Hysterectomy/veterinaryالملخص
Abstract The process of ovulation involves multiple and iterrelated genetic, biochemical, and morphological events: cessation of the proliferation of granulosa cells, resumption of oocyte meiosis, expansion of cumulus cell-oocyte complexes, digestion of the follicle wall, and extrusion of the metaphase-II oocyte. The present narrative review examines these interrelated steps in detail. The combined or isolated roles of the folliclestimulating hormone (FSH) and luteinizing hormone (LH) are highlighted. Genes indiced by the FSH genes are relevant in the cumulus expansion, and LH-induced genes are critical for the resumption ofmeiosis and digestion of the follicle wall. A nonhuman model for follicle-wall digestion and oocyte release was provided.
Resumo O processo de ovulação envolve modificações genéticas, bioquímicas e morfológicas múltiplas e interrelacionadas: suspensão da proliferação das células da granulosa, reinício da meiose do oócito, expansão das células do complexo cumulus-oócito, digestão da parede folicular, e extrusão do oócito. Esta revisão narrativa examina em detalhes cada um desses eventos e os principais genes e proteínas envolvidos. Mais importante, a ação combinada ou isolada do hormônio folículo-estimulante (HFE) e do hormônio luteinizante (HL) é destacada. Detalha-se o papel do HFE na expansão do cumulus e do HL na digestão da parede folicular, permitindo a extrusão do oócito na superfície ovariana. Proveu-se um modelo não humano para explicar a digestão da parede folicular.
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Humans , Animals , Female , Ovulation/physiology , Luteinizing Hormone/physiology , Oocytes/growth & development , Ovulation/genetics , Luteinizing Hormone/genetics , Signal Transduction , Models, Animal , Cumulus Cells/physiology , Follicle Stimulating Hormone/physiology , Follicle Stimulating Hormone/genetics , Ovarian Follicle/growth & development , Granulosa Cells/physiology , Meiosis/physiology , Meiosis/geneticsالملخص
RESUMEN Se estudió la biología reproductiva del Bocachico, Prochilodus magdalenae, en el río San Jorge, Colombia. La especie es un pez con proporción sexual hembra: macho de 1,2:1, diferente a lo esperado. Presenta desarrollo ovocitario sincrónico en dos grupos, un desove anual que se extiende de abril a septiembre asociado al ciclo hidrológico del río San Jorge, talla media de madurez sexual estimada en 30,2 cm LT, ovocitos grandes de 950 μm y fecundidad promedio de 109 972 ovocitos, cuya ecuación fue F =7271,6 WO0,60, r =0,93, n =14.
ABSTRACT The reproductive biology of the Bocachico, Prochilodus magdalenae, in the San Jorge River, Colombia, was studied. The species is a fish with sexual proportion female: male of 1.2:1, different than expected. It presents synchronous ovocitary development in two groups, an annual spawning that extends from April to September associated to the hydrological cycle of the San Jorge River, length at first maturity estimated in 30.2 cm TL, large oocytes of 950 μm, and average fecundity of 109 972 oocytes, whose equation was F = 7271,6 OW0,60, r = 0,93, n = 14.
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@#<p>Conjoined oocytes are characterized by a follicle containing two oocytes surrounded by a single zona pellucida or their fusion in a zonal region. Gonadotropin stimulation, coupled with PCOS, predisposes to the occurrence of conjoined oocytes. Although a result of developmental accident, conjoined oocytes have the potential to develop into genetically normal embryos and babies. In this paper, the authors describe the aspiration of multiple conjoined oocytes in a single IVF cycle, leading to normal fertilization after selective intracytoplasmic sperm injection (ICSI). Development to blastocyst stage allowed trophectoderm biopsy resulting to a euploid embryo that was eventually transferred, and resulted to a live term birth.</p><p>Key words: conjoined oocytes, in vitro fertilization (IVF), intracytoplasmic sperm injection(ICSI)</p>
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Fertilization in Vitro , Sperm Injections, Intracytoplasmicالملخص
Parthenogenetic embryos, created by activation and diploidization of oocytes, arrest at mid-gestation for defective paternal imprints, which impair placental development. Also, viable offspring has not been obtained without genetic manipulation from parthenogenetic embryonic stem cells (pESCs) derived from parthenogenetic embryos, presumably attributable to their aberrant imprinting. We show that an unlimited number of oocytes can be derived from pESCs and produce healthy offspring. Moreover, normal expression of imprinted genes is found in the germ cells and the mice. pESCs exhibited imprinting consistent with exclusively maternal lineage, and higher X-chromosome activation compared to female ESCs derived from the same mouse genetic background. pESCs differentiated into primordial germ cell-like cells (PGCLCs) and formed oocytes following in vivo transplantation into kidney capsule that produced fertile pups and reconstituted ovarian endocrine function. The transcriptome and methylation of imprinted and X-linked genes in pESC-PGCLCs closely resembled those of in vivo produced PGCs, consistent with efficient reprogramming of methylation and genomic imprinting. These results demonstrate that amplification of germ cells through parthenogenesis faithfully maintains maternal imprinting, offering a promising route for deriving functional oocytes and having potential in rebuilding ovarian endocrine function.
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Animals , Female , Mice , Mice, Transgenic , Mouse Embryonic Stem Cells/metabolism , Oocytes/metabolism , Parthenogenesisالملخص
Background: Birth is often referred to as “the miracle of life” and very rightly so since successful pregnancy represents a complex, a highly coordinated succession of biological processes. In this regard, it is remarkable that pregnancy ever occurs at all. Infertility is a reproductive system disorder affecting 10.7% to 15.5% of couples. Highly heterogeneous in etiology. Investigating the cause is necessary for guiding treatment options. Aim of the study was to determine the percentage of immature oocytes retrieved at the metaphase 1 stage and germinal vesicle stage and its outcome.Methods: Total 30 cases were included for this retrospective analysis of data done at ARC fertility center, Saveetha Medical College. This study analyzed the data of patients with unexplained infertility who underwent ICSI centers (affiliated to Saveetha University) meeting inclusion criteria and willing to participate in the study.Results: The mean number of fertilized oocytes among those with more than 25% immature oocyte proportion was 1.18 while among those with less than 25% immature oocyte proportion was 4.13. The chance of fertilization of the oocyte increases when the percentage of immature oocyte was less than or equal to 25%. The association was found to be statistically significant with p-value <0.05.Conclusions: Thus, some intrinsic oocyte factor could be involved in causing maturation arrest in such couples and resulting in multiple failed IVF cycles.so this oocyte factor could be a major cause in many cases of unexplained infertility.
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Background: recently, it has been proved that copolymers with dextran cores and grafted polyacrylamide are effective in photodynamic and chemotherapy. However, further research is needed to define correct dosage and to assess the risks. Thus, animal studies are becoming more relevant to determine the effect of the treatment of such drug nano-systems on female reproductive function in particular.Methods: a technique for estimation of pre- and post-implantation death rates, in vitro meotic maturation of oocytes, double fluorescent vital assay and statistical analysis were used. The effects of a one-time treatment of different doses of dextran-polyacrylamide matrices and silver (Ag)-nanoparticles-dextran-polyacrylamide (AgNPs-D-PAA) on reproductive function, namely on 1) the number of oocytes isolated from one ovary and the meiotic maturation of such oocytes in vitro; 2) the indicators of cell viability of the cells of follicular environment of oocytes (FEO) and the cells of inguinal lymph nodes (ILN); 3) the pre- and post-implantation mortality rates and the number of live newborns (pups) were investigated in female mice.Results: no significant changes in the number of oocytes isolated from one ovary and meiotic maturation of such ovarian oocytes in vitro, the number of living cells of follicular environment of oocytes and the number of such cells with morphological signs of apoptosis and necrosis, pre- and post-implantation mortality rates of embryos and the number of live newborns (pups) have been established under conditions of one-time treatment with dextran-polyacrylamide at doses of 0.39 mg/kg and 3.90 mg/kg and Ag-nanoparticles-dextran-polyacrylamide at doses of 0.20 mg/kg and 2.00 mg/kg.Conclusions: branched polymer systems (dextran-polyacrylamide (D-PAA) polymer matrices) are promising materials for use in next-generation medicine.
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@#Ovarian aging has been associated with oxidative stress and loss of ovarian function. Tocotrienol has been proven to exert beneficial effects on the female reproductive system. However, the role of tocotrienol in affecting metabolism in the ovary and subsequently improving the quality of oocytes in aging mice remains unknown. Therefore, the relationship between metabolic changes in the ovary and the quality of oocytes in aging mice following tocotrienol-rich fraction (TRF) supplementation was investigated. Six-week-old female mice were used as the Young group. Six-month-old aged female mice were divided into four groups; the first group was given tocopherol-stripped corn oil (vehicle control) while the other three groups were supplemented with TRF at the dose of 90, 120, and 150 mg/kg, respectively. The treatment was given orally for two months. At the end of the treatment, mice from all groups were superovulated and then euthanised. Oocyte quality was assessed and non-targeted metabolomic analysis of the ovarian tissues was performed using liquid chromatography-tandem mass spectrometry of quadrupole time-of-flight (LC-MS Q-TOF). Percentages of normal oocytes were higher (p<0.001) while abnormal oocytes were lower (p<0.001) in TRF 150 mg/kg group compared to that of the control. Seventeen metabolites were identified to be significantly different in the ovarian tissue of the aging group when compared to the young group. 14 metabolites were identified to be significantly different in the ovarian tissue between the control and TRF supplemented groups. Pathway analysis revealed significantly altered metabolic pathways for fatty acid and amino acid metabolism that might influence the quality of oocytes. In conclusion, TRF supplementation causes metabolic changes in the ovary that delay the consequences of aging, thus improving the quality of oocytes in aging mice.
الملخص
Resumen El objetivo de este estudio fue evaluar el consumo de agua tratada proveniente de la producción petrolera en la calidad seminal del macho bovino reproductor, para ello se seleccionaron 16 toros entre 4 y 5 años distribuidos en dos grupos en los centros de investigaciones La Libertad de Agrosavia en Villavicencio y el Área de sostenibilidad agroenergetica, ASA en Acacias con disponibilidad de agua tratada de los campos petroleros, Apiay (A) y Castilla (C) respectivamente. Los animales seleccionados fueron distribuidos al azar para cada localidad en 4 tratamientos: 1) Consumo del 100 % agua de producción tratada; 2) consumo de mezcla 50 % agua de producción tratada y 50 % agua cruda; 3) consumo de mezcla 25 % agua de producción tratada y 75 % agua cruda; y 4) consumo 100 % agua cruda. Las variables consideradas fueron: calidad seminal para A y C determinadas por un sistema computarizado y de fertilidad in vitro para A evaluada por tinción fluorescente, Hoechst 33342. Los datos se analizaron estadística descriptiva, análisis de varianza (ANOVA) y pruebas de comparación de medias, Tukey, en un modelo de medidas repetidas en el tiempo. Los resultados indicaron, para calidad seminal, que los tratamientos que consumieron A y C en cada localidad presentaron diferencias (P < 0,05) para motilidad, 75,00±4,50, 69,24±4,13, espermatozoides motiles/100 células evaluadas, en el tratamiento dos respectivamente y en A para morfología, 76,67±2,06 espermatozoides normales/100 en el tratamiento cuatro, el índice de calidad espermática (ICE) no mostró diferencia (P > 0,05). Los resultados de unión espermatozoide - zona pelúcida mostraron diferencias entre tratamientos (P < 0,05), al igual que en fecundación in vitro, sin embargo, el comportamiento de estos cambios no indica asociación con el consumo de este tipo de agua. El estudio muestra que no se observan cambios contundentes o negativos que demuestren efecto en la fertilidad del toro por consumo de agua de producción tratada.
Abstract The objective of this study was to evaluate the consumption of treated water from oil production in the seminal quality of the breeding male; To this end, 16 bulls between 4 and 5 years old were selected, distributed in two groups at Agrosavia's La Libertad centers in Villavicencio and the Agroenergetic Sustainability Area, ASA in Acacias with availability of treated water from the oil fields, Apiay (A) and Castilla (C) respectively. The selected animals were distributed randomly to each location in 4 treatments: 1) Consumption of 100% treated production water; 2) 50% consumption of treated production water and 50% raw water consumption; 3) mixing consumption 25% treated production water and 75% raw water; and 4) 100% raw water consumption. The variables considered were: seminal quality for A and C determined by a computerized system, and in vitro fertility for A evaluated by fluorescent staining, Hoechst 33342. The data were analyzed descriptive statistics, analysis of variance (ANOVA) and means comparison tests, Tukey, in a model of measures repeated over time. The results indicated, for seminal quality, that the treatments that consumed A and C in each locality presented differences (P <0,05) for motility, 75,00 ± 4,50, 69,24 ± 4,13, motile sperm / 100 cells evaluated, in treatment two respectively and in A for morphology, 76,67 ± 2,06 normal sperm / 100 in treatment four, the sperm quality index (SQI) showed no difference (P> 0,05). The results of sperm-pellucid zone binding showed differences between treatments (P <0,05), as in in vitro fertilization, however, the behavior of these changes does not indicate an association with the consumption of this type of water. The study shows that there are no strong or negative changes that show an effect on the fertility of the bull due to the consumption of treated production water.
Resumo O objetivo deste estudo foi avaliar o consumo de água tratada da produção de óleo na qualidade seminal do macho reprodutor; Para tanto, foram selecionados 16 touros entre 4 e 5 anos, distribuídos em dois grupos nos centros de La Libertad de Agrosavia em Villavicencio e na Área de Sustentabilidade Agroenergética, ASA em Acacias com disponibilidade de água tratada dos campos de petróleo, Apiay (A ) e Castilla (C) respectivamente. Os animais selecionados foram distribuídos aleatoriamente para cada local em 4 tratamentos: 1) Consumo de água de produção tratada 100%; 2) 50% de consumo de água de produção tratada e 50% de consumo de água bruta; 3) consumo de mistura 25% de água de produção tratada e 75% de água bruta; e 4) 100% de consumo de água bruta. As variáveis consideradas foram: qualidade seminal para A e C, determinada por sistema computadorizado, e fertilidade in vitro para A, avaliada por coloração fluorescente, Hoechst 33342. Os resultados indicaram, para qualidade seminal, que os tratamentos que consumiram A e C em cada localidade apresentaram diferenças (P <0,05) para motilidade, 75,00 ± 4,50, 69,24 ± 4,13, espermatozóides móveis / 100 células avaliadas, no tratamento dois respectivamente e em A para morfologia, 76,67 ± 2,06 espermatozóides normais / 100 no tratamento quatro, o índice de qualidade espermática (IQE) não apresentou diferença (P> 0,05). Os resultados de ligação das zonas pelúcidas de espermatozóides mostraram diferenças entre os tratamentos (P <0,05), como na fertilização in vitro, entretanto, o comportamento dessas mudanças não indica uma associação com o consumo desse tipo de água. O estudo mostra que não há mudanças fortes ou negativas que mostram um efeito sobre a fertilidade do touro devido ao consumo de água de produção tratada.
الملخص
Abstract Galanthamine is an Amaryllidaceae-derived acetylcholinesterase inhibitor used to treat memory impairment in Alzheimer's disease and vascular dementia. There is evidence that galanthamine, in addition to its effects on acetylcholinesterase, may enhance or inhibit brain nicotinic acetylcholine receptors, which could increase or decrease the therapeutic efficacy of galanthamine, respectively. Here, we evaluated the effects of galanthamine and two others Amaryllidaceae acetylcholinesterase inhibitors (haemanthamine and tazettine) analyzed by gas chromatography-mass spectrometry and identified by comparing their mass fragmentation patterns with literature and database NIST vs.2.0 on the agonist responses of brain nicotinic acetylcholine receptors α7, α3β4, (α4)2(β2)3 and (α4)3(β2)2. Using nicotinic acetylcholine receptors expressed heterologously in Xenopus oocytes, in conjunction with two-electrode voltage clamping, we found that galanthamine inhibits the function of nicotinic acetylcholine receptors assayed through a mix competitive and non-competitevely. Nicotinic acetylcholine receptor α7 were significantly more sensitive to inhibition (17 ± 0.6 µM) than the heteromeric receptor, α3β4 (90 ± 3.4 µM). Neither haemanthamine nor tazettine were more potent than galanthamine.
الملخص
Background: CPEB is considered as an RNA-binding protein first identified in Xenopus oocytes. Although CPEB1 was involved in the growth of oocyte, its role in goat follicular granulosa cell has not been fully elucidated. To clarify the functions of this gene in goat follicular granulosa cells, CPEB1-overexpressing vector and interference vector were structured and transfected into follicular granulosa cells from Jiangsu native white goats of Nantong city, Jiangsu Province, China. The expression levels of differentiation-related genes including CDK1, Cyclin B1, and C-mos were determined 24 h after administration of CPEB1 by quantitative real-time polymerase chain reaction and Western blotting methods. Results: The expression levels of CDK1, Cyclin B1, and C-mos were significantly upregulated after overexpression and significantly downregulated after interference with CPEB1. Conclusions: The CPEB1 gene expression could affect the transcription of genes related to early cleavage divisions, which provided a reference for further research on its role in the growth and maturation of oocytes.
الموضوعات
Animals , Female , Oocytes , Transcription Factors/genetics , Goats/genetics , Transfection , Fertilization in Vitro , Gene Expression , Blotting, Western , Polymerase Chain Reaction/methods , RNA-Binding Proteins , Embryo Transfer , Livestock , Fluorescence , Granulosa Cellsالملخص
OBJECTIVE@#To explore the causes of oocyte vitrification and its application in assisted reproduction.@*METHODS@#We retrospectively analyzed the data of 26 patients with 27 cycles of oocyte vitrification cryopreservation undergoing intracytoplasmic sperm injection (ICSI) and embryo transfer between January, 2008 and October, 2018. The causes of oocyte vitrification and the outcomes of ICSI and clinical pregnancy were analyzed.@*RESULTS@#The causes of oocytes vitrification included mainly azoospermia or severe spermatogenesis disorder of the husband, failure to obtain sperms from the husband, failure of the husband to be present on the day of oocyte retrieval and acute diseases of the husband to not allow sperm collection. A total of 274 oocytes were frozen in 27 oocyte retrieval cycles, and 217 eggs were thawed in 19 cycles with a survival rate of 81.11% (176/217). The normal fertilization rate, cleavage rate and high-quality embryo rate was 74.81% (98/131), 89.80% (88/98) and 36.73% (36/98), respectively. Fifteen patients underwent embryo transfer, and the clinical pregnancy rate and live birth rate was 53.33% (8/15) and 33.33% (5/15), respectively. Compared with patients below 35 years of age, the patients aged above 35 years had significantly lower oocyte survival rate after thawing (82.76% 74.42%, =0.211), clinical pregnancy rate (77.78% 16.67%, =0.041) and live birth rate (55.56% 0, =0.044).@*CONCLUSIONS@#Oocytes vitrification can be used as a remedy for infertile couples who fail to provide sperms due to male factors on the day of oocyte retrieval. Vitrification of the oocytes does not significantly affect the fertilization rate or the clinical pregnancy rate. The survival rate of the thawed oocytes is related to the age of the wife, and an age younger than 35 years can be optimal for achieving favorable clinical pregnancy outcomes after oocyte vitrification.