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1.
Cell ; 168(3): 473-486.e15, 2017 01 26.
Artigo em Inglês | MEDLINE | ID: mdl-28129541

RESUMO

Interspecies blastocyst complementation enables organ-specific enrichment of xenogenic pluripotent stem cell (PSC) derivatives. Here, we establish a versatile blastocyst complementation platform based on CRISPR-Cas9-mediated zygote genome editing and show enrichment of rat PSC-derivatives in several tissues of gene-edited organogenesis-disabled mice. Besides gaining insights into species evolution, embryogenesis, and human disease, interspecies blastocyst complementation might allow human organ generation in animals whose organ size, anatomy, and physiology are closer to humans. To date, however, whether human PSCs (hPSCs) can contribute to chimera formation in non-rodent species remains unknown. We systematically evaluate the chimeric competency of several types of hPSCs using a more diversified clade of mammals, the ungulates. We find that naïve hPSCs robustly engraft in both pig and cattle pre-implantation blastocysts but show limited contribution to post-implantation pig embryos. Instead, an intermediate hPSC type exhibits higher degree of chimerism and is able to generate differentiated progenies in post-implantation pig embryos.


Assuntos
Quimerismo , Edição de Genes , Mamíferos/embriologia , Animais , Blastocisto , Sistemas CRISPR-Cas , Bovinos , Embrião de Mamíferos/citologia , Feminino , Humanos , Masculino , Mamíferos/classificação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos ICR , Células-Tronco Pluripotentes , Ratos , Ratos Sprague-Dawley , Sus scrofa
2.
Reprod Domest Anim ; 57 Suppl 5: 86-89, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35467050

RESUMO

Seminal plasma (SP) affects reproduction, inducing cell and molecular changes in the female genital tract. A main active component in SP is the modulatory transforming growth factor-ß (TGF-ß), particularly its TGF-ß1 isoform, which affects the synthesis of other cytokines as granulocyte-macrophage colony-stimulating factor, relevant for embryo development and pregnancy. This study evaluated the effect of pooled frozen-thawed SP and commercial TGF-ß1 infused during oestrus in sows post-cervically inseminated with liquid extended semen, containing ~4 ml of residual SP, on their fertility and prolificacy. For this, 250 sows in their post-weaning oestrus were used. Sows were randomly assigned to one of the following groups to be post-cervically treated 30 min before insemination: (i) SP group: infused with 40 ml of SP (N = 57); ii) Group TFGß1 : infused with 40 ml of BTS extender containing 3 ng/ml of porcine TGF-ß1 (N = 64); iii) BTS group: infused with 40 ml of BTS extender (N = 60); and iv) Control Group: sows catheterized but not infused prior to AI (N = 69). Farrowing rates (range: 86.7% to 91.3%) and numbers of live-born piglets (range: range: 12.8 ± 2.9 to 13.4 ± 3.1) were not affected by any treatment compared with Controls, indicating that neither pre-infusions of SP nor TGF-ß1 30 min before AI influenced subsequent fertility and prolificacy.


Assuntos
Preservação do Sêmen , Sêmen , Animais , Criopreservação/veterinária , Citocinas , Feminino , Fertilidade , Fator Estimulador de Colônias de Granulócitos e Macrófagos , Inseminação Artificial/veterinária , Masculino , Gravidez , Preservação do Sêmen/veterinária , Espermatozoides , Suínos , Fator de Crescimento Transformador beta1/farmacologia , Fatores de Crescimento Transformadores
3.
Reprod Domest Anim ; 57 Suppl 5: 4-13, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35500162

RESUMO

The establishment of a successful pregnancy can only occur through a concerted functioning of the entire female reproductive system, allowing for fertilization, subsequent embryo development and implantation of the conceptus. In this context, the uterine immunological responses responsible for rejection or tolerance of the conceptus are of critical importance. The aim of the present review is to summarize our current knowledge about those cellular and molecular immunological events occurring at the uterine level during pre-implantation and implantation stages of pregnancy in the pig. Advancing our understanding of the immune mechanisms involved in the success or failure of pregnancy will provide cues to develop novel strategies augmenting endometrial receptivity, finally increasing the efficiency of assisted reproductive technologies in pigs.


Assuntos
Implantação do Embrião , Útero , Animais , Implantação do Embrião/fisiologia , Embrião de Mamíferos , Desenvolvimento Embrionário , Endométrio , Feminino , Gravidez , Suínos , Útero/fisiologia
4.
Reprod Domest Anim ; 57 Suppl 5: 58-63, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-35567517

RESUMO

The aims of this study were to investigate the effects of different equilibration times with cryoprotectants on viability and metaphase plate morphology of vitrified-warmed porcine mature oocytes (Experiment 1) and to evaluate the effects of supplementation with 10-9 M melatonin during in vitro maturation on these parameters (Experiment 2). In Experiment 1, 2,392 mature oocytes were vitrified using different equilibration times of oocytes with cryoprotectants (3, 10, 15, 20, 30, 40, 60 and 80 min). Fresh oocytes matured in vitro for 44 hr (n = 509) were used as controls. In Experiment 2, a total of 573 COCs were used. COCs were matured with 10-9 M melatonin supplementation or without melatonin (control). Some oocytes from each group were vitrified with a 60-min equilibration time with cryoprotectants according to the results of Experiment 1. The remaining oocytes from each maturation group were used as fresh control groups. In both experiments, oocytes were stained with 2',7'-dichlorodihydrofuorescein diacetate and Hoechst 33342 to assess viability and metaphase plate morphology, respectively. Vitrification and warming affected (p < .01) oocyte viability compared with controls, which were all viable after 44 hr of IVM. In Experiment 1, the longer the equilibration time with cryoprotectants, the higher the viability. Oocytes equilibrated for 60 and 80 min had the highest (p < .05) viability and similar metaphase plate characteristics to the fresh control oocytes. In Experiment 2, supplementation with melatonin during in vitro maturation had no effect on oocyte viability or metaphase plate morphology of vitrified-warmed oocytes. In conclusion, under our experimental conditions, vitrified porcine mature oocytes equilibrated with cryoprotectants for 60 or 80 min exhibited the highest viability and similar metaphase plate characteristics to fresh controls. Furthermore, supplementation with 10-9 M melatonin during in vitro maturation had no effect on these parameters.


Assuntos
Melatonina , Animais , Criopreservação/métodos , Criopreservação/veterinária , Crioprotetores/farmacologia , Suplementos Nutricionais , Melatonina/farmacologia , Metáfase , Oócitos , Suínos , Vitrificação
5.
Reprod Domest Anim ; 54 Suppl 4: 72-77, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31625244

RESUMO

The coenzyme Q10 (CoQ10) is a potent antioxidant with critical protection role against cell oxidative stress, caused by the mitochondrial dysfunction. This study evaluated the effects of CoQ10 supplementation to in vitro maturation (IVM) or embryo culture media on the maturation, fertilization and subsequent embryonic development of pig oocytes and embryos. Maturation (Experiment 1) or embryo culture (Experiment 2) media were supplemented with 0 (control), 10, 25, 50 and 100 µM CoQ10. The addition of 10-50 µM CoQ10 to the IVM medium did not affect the percentage of MII oocytes nor the fertilization or the parameters of subsequent embryonic development. Exogenous CoQ10 in the culture medium neither did affect the development to the 2-4-cell stage nor rates of blastocyst formation. Moreover, the highest concentration of CoQ10 (100 µM) in the maturation medium negatively affected blastocyst rates. In conclusion, exogenous CoQ10 supplementation of maturation or embryo culture media failed to improve the outcomes of our in vitro embryo production system and its use as an exogenous antioxidant should not be encouraged.


Assuntos
Técnicas de Cultura Embrionária/veterinária , Técnicas de Maturação in Vitro de Oócitos/veterinária , Ubiquinona/análogos & derivados , Animais , Antioxidantes/farmacologia , Meios de Cultura/química , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário/efeitos dos fármacos , Feminino , Fertilização in vitro/veterinária , Técnicas de Maturação in Vitro de Oócitos/métodos , Masculino , Oócitos/efeitos dos fármacos , Suínos , Ubiquinona/efeitos adversos , Ubiquinona/farmacologia
6.
Reprod Domest Anim ; 54 Suppl 4: 4-13, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31625238

RESUMO

Commercial embryo transfer (ET) has unprecedented productive and economic implications for the pig sector. However, pig ET has been considered utopian for decades mainly because of the requirements of surgical techniques for embryo collection and embryo deposition into recipients, alongside challenges to preserve embryos. This situation has drastically changed in the last decade since the current technology allows non-surgical ET and short- and long-term embryo preservation. Here, we provide a brief review of the improvements in porcine ET achieved by our laboratory in the past 20 years. This review includes several aspects of non-surgical ET technology and different issues affecting ET programmes and embryo preservation systems. The future perspectives of ET technology are also considered. We will refer only to embryos produced in vivo since they are the only type of embryos with possible short-term use in pig production.


Assuntos
Transferência Embrionária/veterinária , Suínos/embriologia , Animais , Criopreservação/métodos , Criopreservação/veterinária , Técnicas de Cultura Embrionária/veterinária , Transferência Embrionária/métodos , Embrião de Mamíferos , Feminino
7.
BMC Genomics ; 15: 293, 2014 May 21.
Artigo em Inglês | MEDLINE | ID: mdl-24886317

RESUMO

BACKGROUND: Sex allocation of offspring in mammals is usually considered as a matter of chance, being dependent on whether an X- or a Y-chromosome-bearing spermatozoon reaches the oocyte first. Here we investigated the alternative possibility, namely that the oviducts can recognise X- and Y- spermatozoa, and may thus be able to bias the offspring sex ratio. RESULTS: By introducing X- or Y-sperm populations into the two separate oviducts of single female pigs using bilateral laparoscopic insemination we found that the spermatozoa did indeed elicit sex-specific transcriptomic responses. Microarray analysis revealed that 501 were consistently altered (P-value < 0.05) in the oviduct in the presence of Y-chromosome-bearing spermatozoa compared to the presence of X-chromosome-bearing spermatozoa. From these 501 transcripts, 271 transcripts (54.1%) were down-regulated and 230 transcripts (45.9%) were up-regulated when the Y- chromosome-bearing spermatozoa was present in the oviduct. Our data showed that local immune responses specific to each sperm type were elicited within the oviduct. In addition, either type of spermatozoa elicits sex-specific signal transduction signalling by oviductal cells. CONCLUSIONS: Our data suggest that the oviduct functions as a biological sensor that screens the spermatozoon, and then responds by modifying the oviductal environment. We hypothesize that there might exist a gender biasing mechanism controlled by the female.


Assuntos
Oviductos/fisiologia , Processos de Determinação Sexual , Espermatozoides/metabolismo , Transcriptoma , Cromossomo X , Cromossomo Y , Animais , Feminino , Masculino , Suínos
8.
J Reprod Dev ; 60(5): 371-6, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25030061

RESUMO

This study aimed to evaluate the effect of recipient-donor estrous cycle synchrony on recipient reproductive performance after nonsurgical deep-uterine (NsDU) embryo transfer (ET). The transfers (N=132) were conducted in recipients sows that started estrus 24 h before (-24 h; N=9) or 0 h (synchronous; N=31), 24 h (+24 h; N=74) or 48 h (+48 h; N=18) after the donors. A total of 30 day 5 morulae or day 6 blastocysts (day 0=onset of estrus) were transferred per recipient. The highest farrowing rates (FRs) were achieved when estrus appeared in recipients 24 h later than that in the donors (81.1%), regardless of the embryonic stage used for the transfers. The FR notably decreased (P<0.05) when recipients were -24 h asynchronous (0%), synchronous (61.3%) or +48 h asynchronous (50%) relative to the donors. No differences in litter size (LS) and piglet birth weights were observed among the synchronous and +24 h or +48 h asynchronous groups. While a +24 h asynchronous recipient was suitable for transfers performed with either morulae (FR, 74.3%; LS, 9.2 ± 0.6 piglets) or blastocysts (FR, 84.6%; LS, 9.8 ± 0.6 piglets), a + 48 h asynchronous recipient was adequate for blastocysts (FR, 87.5%; LS, 10.4 ± 0.7 piglets) but not for morulae (FR, 30.0%; LS, 7.3 ± 2.3 piglets). In conclusion, this study confirms the effectiveness of the NsDU-ET technology and shows that porcine embryos tolerate better a less advanced uterine environment if they are nonsurgically transferred deep into the uterine horn.


Assuntos
Transferência Embrionária/veterinária , Desenvolvimento Embrionário/fisiologia , Estro/fisiologia , Sus scrofa , Útero/fisiologia , Animais , Blastocisto/fisiologia , Transferência Embrionária/métodos , Sincronização do Estro/fisiologia , Feminino , Mórula/fisiologia , Gravidez
9.
Mol Reprod Dev ; 79(9): 651-63, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22777700

RESUMO

Hoechst 33342 (H342), in combination with ultraviolet (UV) irradiation, is frequently used to aid or confirm the enucleation of porcine oocytes in somatic cell nuclear transfer programs. The exposure of oocytes to H342 and UV irradiation has a deleterious effect on the development of in vitro-fertilized porcine oocytes, with increasing exposure to UV irradiation (up to 30 sec) having more drastic effects. It has been hypothesized that this decrease in embryonic development could be due to damage to the mitochondrial DNA (mtDNA). To investigate this hypothesis, we analyzed the mitochondrial distribution and DNA copy number of in vitro-matured porcine oocytes exposed to H342/UV and the subsequent embryonic development compared with the mitochondrial distribution and DNA copy number of in vivo-derived oocytes and embryos. Using quantitative, real-time polymerase chain reaction (qPCR) protocols to analyze mtDNA and confocal laser scanning microscopy with MitoTracker Deep Red to determine mitochondrial distribution, we demonstrated that the simultaneous exposure of in vitro-matured porcine oocytes to H342 staining and UV irradiation is associated with reduced oocyte developmental competence and abnormal mitochondrial distribution in the resulting cleaved embryos. In addition, 2- to 4-cell embryos derived from oocytes exposed to H342/UV showed a significant decrease in mtDNA copy number. These results should be considered when H342/UV procedure is used during nuclear transfer in recipient porcine oocytes.


Assuntos
Benzimidazóis/efeitos adversos , Blastocisto/metabolismo , Variações do Número de Cópias de DNA , DNA Mitocondrial/metabolismo , Corantes Fluorescentes/efeitos adversos , Mitocôndrias/metabolismo , Oócitos/metabolismo , Raios Ultravioleta/efeitos adversos , Animais , Benzimidazóis/farmacologia , Blastocisto/patologia , Variações do Número de Cópias de DNA/efeitos dos fármacos , Variações do Número de Cópias de DNA/efeitos da radiação , Feminino , Corantes Fluorescentes/farmacologia , Mitocôndrias/patologia , Oócitos/patologia , Reação em Cadeia da Polimerase em Tempo Real , Suínos , Fatores de Tempo
10.
Anim Reprod ; 17(3): e20200022, 2020 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-33029213

RESUMO

Seminal plasma (SP) supports not only sperm function but also the ability of spermatozoa to withstand biotechnological procedures as artificial insemination, freezing or sex sorting. Moreover, evidence has been provided that SP contains identifiable molecules which can act as fertility biomarkers, and even improve the output of assisted reproductive technologies by acting as modulators of endometrial and embryonic changes of gene expression, thus affecting embryo development and fertility beyond the sperm horizon. In this overview, we discuss current knowledge of the composition of SP, mainly proteins and cytokines, and their influence on semen basic procedures, such as liquid storage or cryopreservation. The role of SP as modulator of endometrial and embryonic molecular changes that lead to successful pregnancy will also be discussed.

11.
Rev. Finlay ; 11(1): 51-58, ene.-mar. 2021. tab, graf
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1250791

RESUMO

RESUMEN Fundamento: los defectos congénitos se ubican entre las primeras causas de muerte en menores de un año. Constituyen un problema para el individuo, la familia, y la sociedad. En Cuba constituyenlaprimera causa de muerte en menores de un año y en Cienfuegos ocupaneltercer lugar. Objetivo: describir los defectos congénitos más frecuentes presentados en la provincia Cienfuegos entre los años 2011 y 2018 y determinar la prevalencia al nacer, frecuencia ajustada yla tendencia. Métodos: se realizó un estudio descriptivo, de corte transversal de los defectos congénitos más frecuentes en la provincia Cienfuegos, en el período 2011- 2018. El universo quedó constituido por330 niños nacidos vivos con diagnóstico de defectos congénitos y 286 terminaciones voluntarias de la gestación. Los resultados se presentaron en tablas o gráficos según fue más factible mostrar la información. Las variables analizadas fueron: prevalencia al nacer por año de la ocurrencia de defectos congénitos, frecuencia ajustada y anomalías mayores más frecuentes en la provincia.El análisis de los datos se realizó mediante la recogida de información en una base de datos para lo que se utilizó el modelo del Registro Cubano de Malformaciones Congénitas y el Registro Cubano Prenatal de Malformaciones Congénitas. Se determinó la prevalencia al nacer y la frecuencia ajustada, así como la tendencia en la ocurrencia de algunos defectos congénitos en la provincia. Los resultados se presentaron en gráficos y tablassegún fuera más factible mostrar la información Resultados: el síndrome de Down, la hidrocefalia y los defectos de abiertos de pared anterior fueron los que con más frecuencia se presentaron en la población cienfueguera, con una tendencia al decrecimiento en la aparición de anomalías como: atresia esofágica, transposición de grandes vasos,entre otros. Conclusiones: el síndrome de Down, la hidrocefalia y los defectos de abiertos de pared anterior fueron los defectos congénitos que con más frecuencia se presentaron en la población pediátrica en Cienfuegos.


ABSTRACT Background: congenital defects are among the leading causes of death in children under one year of age. They constitute a problem for the individual, the family, and society. In Cuba they are the leading cause of death in children under one year of age and in Cienfuegos they rank third. Objective: to describe the most frequent congenital defects presented in the Cienfuegos province between the years 2011 and 2018 and to determine the prevalence at birth, adjusted frequency and trend. Methods: a descriptive, cross-sectional study of the most frequent congenital defects in the Cienfuegos province was carried out, in the period 2011-2018. The universe of 330 live-born children with congenital defects diagnosis and 286 voluntary terminations of pregnancy. The results were presented in data tables or graphs as it was more feasible to display the information. The variables analyzed were: prevalence at birth per year of the occurrence of congenital defects, adjusted frequency, and the most frequent major anomalies in the province. Data analysis was carried out by collecting information in a database for which the model of the congenital malformations Cuban registry and the Cuban congenital malformations prenatal registry was used. The prevalence at birth and the adjusted frequency were determined, as well as the trend in the occurrence of some congenital defects in the province. The results were presented in graphs and data tables as it was more feasible to show the information. Results: down syndrome, hydrocephalus and open anterior wall defects were the ones that most frequently occurred in the Cienfuegos population, with a decreasing trend in the appearance of anomalies such as: esophageal atresia, transposition of the great vessels, among others. Conclusions: down syndrome, hydrocephalus and open anterior wall defects were the congenital defects that most frequently occurred in the pediatric population in Cienfuegos.

12.
Theriogenology ; 63(2): 536-47, 2005 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-15626415

RESUMO

The use of AI in pigs has dramatically expanded in the last few years. New methodological advances in AI are required to serve the requirements of new sperm technologies, such as the use of low dose AI, because the use of cervical AI has a very low efficiency leading to low fertility results. One of the strategies devised to meet these requirements is the deposition of semen near the site of fertilization in the oviduct. Using deep intrauterine insemination with a specially designed catheter, a 20-fold reduction in the number of freshly and diluted inseminated spermatozoa can be achieved without decreasing farrowing rates. Moreover, an advantage of deep intrauterine insemination is the possibility of using processed, 'weaker' spermatozoa such as those that have been frozen-thawed or sex-sorted. Although deep intrauterine insemination should be of benefit to the pig industry, more investigations are needed to understand the mechanisms related to sperm colonization of the oviducts and identify the minimal sperm numbers needed to obtain maximal fertility results for processed and unprocessed boar spermatozoa.


Assuntos
Inseminação Artificial/veterinária , Espermatozoides/fisiologia , Suínos , Útero , Animais , Feminino , Fertilidade , Fertilização , Histeroscopia/veterinária , Inseminação Artificial/métodos , Tamanho da Ninhada de Vivíparos , Masculino , Gravidez , Contagem de Espermatozoides , Útero/anatomia & histologia
13.
Theriogenology ; 59(7): 1605-14, 2003 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-12559465

RESUMO

The present study was carried out to determine the pregnancy rates, farrowing rates and litter size in sows with either induced or spontaneous ovulation inseminated with flow cytometric sorted spermatozoa using deep intrauterine insemination technology. Spermatozoa were stained with Hoechst 33342 and sorted by flow cytometry/cell sorting but not separated into separate X and Y populations. In Experiment 1, sows (n=200) were weaned and treated for estrus/ovulation induction with eCG/hCG. Inseminations with either sorted (70 or 140 million) or non-sorted (70 or 140 million) spermatozoa were done using a specially designed flexible catheter. Farrowing rates were 39.1 and 78.7% for 70 million of sorted and non-sorted, respectively, and 46.6 and 85.7% for 140 million of sorted and non-sorted, respectively (P<0.05). The litter size in sows inseminated with sorted spermatozoa showed a tendency to be lower than when non-sorted spermatozoa were inseminated. In Experiment 2, sows (n=140) were inseminated as in Experiment 1 except that natural estrus was used. The ovaries of these sows were evaluated by transrectal ultrasonography. Farrowing rates were 25 and 77.2% for 70 million of sorted and non-sorted, respectively, and 32 and 80.9% for 140 million of sorted and non-sorted, respectively (P<0.05). These results show that the Deep Intrauterine Insemination technology can be successfully used to produce piglets from sorted spermatozoa when sows are hormonally treated to induce synchronous post weaning oestrus and ovulation.


Assuntos
Inseminação Artificial/veterinária , Tamanho da Ninhada de Vivíparos , Espermatozoides/citologia , Suínos/fisiologia , Animais , Cateterismo/veterinária , Sincronização do Estro , Feminino , Citometria de Fluxo/veterinária , Inseminação Artificial/métodos , Masculino , Gravidez , Resultado da Gravidez/veterinária , Taxa de Gravidez , Contagem de Espermatozoides/veterinária
14.
Theriogenology ; 60(4): 767-76, 2003 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-12832024

RESUMO

The present study was designed to determine the effect of the volume of medium (VM) and the number of oocytes (NOOC) during in vitro fertilization (IVF) on embryo development in pigs. Groups of 15, 30 and 50 in vitro matured oocytes were transferred to 2, 1 and 0.1 ml of modified Tris-buffered medium (mTBM) and inseminated with frozen-thawed spermatozoa (2000 spermatozoa/oocyte) in a 3 x 3 factorial experiment. A total of 2739 oocytes from four replicates were exposed to spermatozoa for 6 h and then cultured in embryo culture medium for 6 h (pronuclear formation) or 7 days (blastocyst formation: BF). The efficiency of fertilization (EF: number of monospermic oocytes/total number of inseminated oocytes) and BF decreased (P<0.03) as the VM increased (EF: 45.9+/-2.2, 43.8+/-2.6 and 36.9+/-1.6% and BF: 29.4+/-2.7, 23.2+/-1.8 and 19.9+/-2.1% for VM 0.1, 1 and 2 ml, respectively). The BF, but not EF, was also affected (P<0.04) by NOOC (19.8+/-1.6, 28.1+/-2.3 and 24.6+/-2.9% for groups of 15, 30 and 50 oocytes, respectively). The effect of the interaction VM x NOOC on EF and BF was not significant. These results indicate that when 2000 spermatozoa/oocyte were used, a low volume of IVF medium (0.1 ml) and the number of oocytes during IVF (30-50) can improve the in vitro embryo production in pigs.


Assuntos
Desenvolvimento Embrionário e Fetal , Fertilização in vitro/veterinária , Oócitos , Suínos/embriologia , Animais , Blastocisto/fisiologia , Contagem de Células , Meios de Cultura , Técnicas de Cultura , Feminino , Trometamina
15.
PLoS One ; 9(8): e104696, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25118944

RESUMO

Excellent fertility and prolificacy have been reported after non-surgical deep uterine transfers of fresh in vivo-derived porcine embryos. Unfortunately, when this technology is used with vitrified embryos, the reproductive performance of recipients is low. For this reason and because the embryos must be stored until they are transferred to the recipient farms, we evaluated the potential application of non-surgical deep uterine transfers with in vivo-derived morulae cultured for 24 h in liquid stage. In Experiment 1, two temperatures (25 °C and 37 °C) and two media (one fully defined and one semi-defined) were assessed. Morulae cultured in culture medium supplemented with bovine serum albumin and fetal calf serum at 38.5 °C in 5% CO2 in air were used as controls. Irrespective of medium, the embryo viability after 24 h of culture was negatively affected (P<0.05) at 25 °C but not at 37 °C compared with the controls. Embryo development was delayed in all experimental groups compared with the control group (P<0.001). Most of the embryos (95.7%) cultured at 37 °C achieved the full or expanded blastocyst stage, and unlike the controls, none of them hatched at the end of culture. In Experiment 2, 785 morulae were cultured in the defined medium at 37 °C for 24 h, and the resulting blastocysts were transferred to the recipients (n = 24). Uncultured embryos collected at the blastocyst stage (n = 750) were directly transferred to the recipients and used as controls (n = 25). No differences in farrowing rates (91.7% and 92.0%) or litter sizes (9.0 ± 0.6 and 9.4 ± 0.8) were observed between the groups. This study demonstrated, for the first time, that high reproductive performance can be achieved after non-surgical deep uterine transfers with short-term cultured morulae in a defined medium, which opens new possibilities for the sanitary, safe national and international trade of porcine embryos and the commercial use of embryo transfer in pigs.


Assuntos
Cruzamento/métodos , Meios de Cultura/química , Técnicas de Cultura Embrionária/veterinária , Transferência Embrionária/veterinária , Mórula/fisiologia , Suínos/fisiologia , Análise de Variância , Animais , Técnicas de Cultura Embrionária/métodos , Transferência Embrionária/métodos , Feminino , Gravidez , Temperatura
16.
Rev. cuba. endocrinol ; 22(3): 210-224, sep.-dic. 2011.
Artigo em Espanhol | LILACS, CUMED | ID: lil-615046

RESUMO

Introducción: las complicaciones metabólicas más importantes para la morbilidad y mortalidad que se derivan de la obesidad tienen como común denominador la resistencia a la insulina. Objetivo: determinar la existencia de resistencia a la insulina e historia familiar de diabetes en un grupo de niños y adolescentes obesos con y sin acantosis nigricans. Métodos: se estudiaron 46 sujetos obesos con y sin acantosis nigricans (21 varones y 25 hembras), con edades entre 4 y 16 años, procedentes de la consulta de endocrinología del Hospital Pediátrico Docente William Soler, en el período de noviembre de 2006 a febrero de 2007. Se les realizó, además de examen físico y anamnesis, prueba de tolerancia a la glucosa con determinación de glucemia e insulinemia en ayunas y a las 2 h. Se utilizó como criterio de resistencia a la insulina el índice HOMA. Resultados: la mayoría de los pacientes (36 sujetos) presentaron resistencia a la insulina, independientemente de la presencia o no de acantosis nigricans, que no estuvo relacionada con el grado de obesidad ni con el pliegue tricipital de grasa, pero sí con la historia familiar de diabetes tipo 2. La presencia de acantosis nigricans estuvo relacionada con el grado de obesidad. Se encontró un 13 por ciento de pacientes con criterios de prediabetes. Conclusiones: la obesidad y la historia familiar de diabetes tipo 2 en los niños y adolescentes se relacionan con la presencia de resistencia a la insulina, independientemente de la presencia de acantosis nigricans(AU)


Introduction: the more important metabolic complications for morbidity and mortality derived from obesity have in common the insulin resistance. Objective: to determine the insulin resistance and the family history of diabetes in a group of obese children and adolescents with and without acanthosis nigricans. Methods: forty six obese subjects with and without acanthosis nigricans (21 boys and 25 girls) aged between 4 and 16, from the William Soler Teaching Children Hospital were studied from November, 2006 to February, 2007. Also, they underwent a physical examination and anamnesis, a test of glucose tolerance with determination of fasting glycemia and insulinemia and at 12 hours. As criterion of insulin resistance the HOMA index was used. Results: most of patients (36 subjects) had insulin resistance, independently of the acanthosis nigricans presence, which was neither related to the obesity degree nor the fat tricipital fold, but yes to the family history of type 2 diabetes. The presence of acanthosis nigricans was related to the obesity degree. There was a 13 percent of patients with pre-diabetes criteria. Conclusions: the obesity and the family history of type 2 diabetes in children and adolescents were related to presence of insulin resistance, independently of presence of acanthosis nigricans(AU)


Assuntos
Humanos , Criança , Adolescente , Resistência à Insulina/fisiologia , Acantose Nigricans/etiologia , Obesidade/metabolismo , Diabetes Mellitus/genética , Teste de Tolerância a Glucose/métodos , Anamnese
17.
Rev. cuba. pediatr ; 80(2)abr.-jun. 2008. tab
Artigo em Espanhol | LILACS | ID: lil-505482

RESUMO

Se presentan 5 pacientes menores de 12 años, atendidos en el Servicio de Endocrinología del Hospital Pediátrico Docente ®William Soler¼ a quienes se les realizó un estudio imagenológico con tomografía axial computada y resonancia magnética nuclear, para confirmar diagnóstico de endocrinopatía. Tras el estudio se diagnosticó síndrome de la silla turca vacía. Existen diferencias clínicas entre niños y adultos con silla turca vacía; las más relevantes radican en que en los niños no hay predominio de sexo ni asociación con la obesidad, y en ellos se observa la presencia de afecciones endocrinas como expresión de alteraciones hipotálamo-hipofisarias.


The cases of 5 patients under 12 that received attention at the Endocrinology Service of William Soler Pediatric Teaching Hospital and that underwent an imaging study with computerized axial tomography and nuclear magnetic imaging to confirm endocrinopathy were presented. The empty sella syndrome was diagnosed after concluding the study. Clinical differences were observed between children and adults with empty sella syndrome. The most significant were found in children, among whom there was neither predominance of sex nor association with obesity. The presence of endocrine affections as expression of hypothalamus-hypophyseal alterations was observed in them.


Assuntos
Humanos , Criança , Adolescente , Imageamento por Ressonância Magnética/métodos , Síndrome da Sela Vazia/epidemiologia , Relatos de Casos
18.
Rev. cuba. obstet. ginecol ; 27(3): 191-198, sept.-dic. 2001. tab
Artigo em Espanhol | LILACS | ID: lil-327257

RESUMO

Se sabe que la ginecología infanto-juvenil es parte de la práctica diaria del ginecólogo. Se comenzó en Cienfuegos, luego de la capacitación de varios especialistas en Ciudad de La Habana, bajo la dirección del Dr. Jorge Peláez. En este trabajo se analizaron los resultados alcanzados en el trienio 1997-1999. Se utilizó el método descriptivo correlacional para determinar el número de pacientes atendidas en la Consulta de ginecología infanto-juvenil. Se observó incremento del número de casos vistos (+ 121) en el año 1998 el cual decreció ligeramente en 1999 con mayor número de consultas ofertadas. Se precisaron los principales motivos de consulta: leucorrea y vulvovaginitis, en primer lugar (26,8 porciento) trastornos menstruales (el 15,1 porciento) en segundo lugar y las aglutinaciones labiales, en tercero (12,3 porciento). Se descartaron las malformaciones genitales y la planificación familiar. Se analizaron los métodos contraceptivos utilizados por nuestros adolescentes: los de barrera, las tabletas contraceptivas y los dispositivos intrauterinos fueron los más usados. Se mostró además, el trabajo relacionado con la interrupción de la gestación diferenciada a adolescentes y se halló reducción sustancial de las mismas, pero se incrementaron las regulaciones menstruales lo que opaca en parte la labor preventiva con el aborto


Assuntos
Humanos , Adolescente , Feminino , Gravidez , Adolescente , Criança , Consultórios Médicos/estatística & dados numéricos , Anticoncepcionais Femininos , Doenças dos Genitais Femininos , Unidade Hospitalar de Ginecologia e Obstetrícia/estatística & dados numéricos
19.
Rev. cuba. oftalmol ; 14(1): 42-46, ene.-jun. 2001. ilus
Artigo em Espanhol | LILACS | ID: lil-331403

RESUMO

Presentamos un caso clínico de displasia oculoauriculovertebral o síndrome de Goldenhar en una paciente femenina de 26 años con múltiples anomalías congénitas y la presencia de quiste branquial y glaucoma primario de ángulo abierto. Mediante un estudio multidisciplinario, clínico, genético, imagenológico y con pruebas psicofísicas, se ponen en evidencia diversas anomalías craneofaciales, óseas y de vísceras toracoabdominales presentes en una misma paciente. Se señalan las diferentes deformidades que conforman actualmente un amplio espectro de anomalías, según la literatura revisada, así como las características clínicas, tratamiento y su evolución. El quiste branquial resulta un hallazgo novedoso que bien puede explicarse en este tipo de alteraciones morfogenéticas, sin embargo, el glaucoma parece ser una entidad casual. Se descartan otras anomalías oculares propias del Goldenhar que pueden ocasionar glaucoma. Se comenta la importancia del diagnóstico y tratamiento multidisciplinario, así como el riesgo quirúrgico de estos enfermos


Assuntos
Humanos , Adulto , Feminino , Anormalidades Múltiplas , Branquioma , Manifestações Oculares , Glaucoma de Ângulo Aberto , Síndrome de Goldenhar/diagnóstico , Síndrome de Goldenhar/genética
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