Allele-Specific Expression of the MAOA Gene and X Chromosome Inactivation in In Vitro Produced Bovine Embryos

During embryogenesis, one of the two X chromosomes is inactivated in embryos. The production of embryos in vitro may affect epigenetic mechanisms that could alter the expression of genes related to embryo development and X chromosome inactivation (XCI). The aim of this study was to understand XCI during in vitro, pre-implantation bovine embryo development by characterizing the allele-specific expression pattern of the X chromosome-linked gene, monoamine oxidase A (MAOA). Two pools of ten embryos, comprised of the 4-, 8- to 16-cell, morula, blastocyst, and expanded blastocyst stages, were collected. Total RNA from embryos was isolated, and the RT-PCR-RFLP technique was used to observe expression of the MAOA gene. The DNA amplicons were also sequenced using the dideoxy sequencing method. MAOA mRNA was detected, and allele-specific expression was identified in each pool of embryos. We showed the presence of both the maternal and paternal alleles in the 4-, 8-to 16-cell, blastocyst and expanded blastocyst embryos, but only the maternal allele was present in the morula stage. Therefore, we can affirm that the paternal X chromosome is totally inactivated at the morula stage and reactivated at the blastocyst stage. To our knowledge, this is the first report of allele-specific expression of an X-linked gene that is subject to XCI in in vitro bovine embryos from the 4-cell to expanded blastocyst stages. We have established a pattern of XCI in our in vitro embryo production system that can be useful as a marker to assist the development of new protocols for in vitro embryo production. Mol. Reprod. Dev. MoL Reprod. Dev. 77: 615-621, 2010. (C) 2010 Wiley-Liss, Inc.

Increased basal FSH levels as predictors of low-quality follicles in infertile women with endometriosis

Objective: To determine whether basal levels of follicle-stimulating hormone (FSH) and anti-mullerian hormone (AMH), antral follicle count (AFC), and the numbers of dominant follicles, oocytes, and mature oocytes retrieved after ovarian stimulation differed between infertile women with endometriosis and healthy women undergoing assisted reproduction techniques (ART). Method: Of 77 consecutive ART candidates, 27 were infertile and had endometriosis. A male factor caused the infertility of the other 50, who acted as controls. Results: The AMH and AFC levels were similar in the 2 groups. The FSH levels were higher (8.28 mIU/ML [range, 5.25-24.1 mIU/ML] vs 5.91 mIU/mL [range, 2.47-18.7 mIU/ML]; P<0.01) in the study group. And the numbers of retrieved (n = 5 [range, 0-12] vs n = 9 [range, 0-27]; P<0.05) and mature oocytes (n = 4 [range, 0-11] vs n = 5 [range, 0-16]; P<0.05) were less in the study group. Conclusion: Because AMH levels were unchanged, endometriosis seems not to damage the primordial pool of follicles and oocytes, but to lessen the quality of the ovarian response to the hCG injection. Basal FSH levels may be of value in predicting ART success in women with the disease. (C) 2010 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.

Supplemented tissue culture medium 199 is a better medium for in vitro maturation of oocytes from women with polycystic ovary syndrome women than human tubal fluid

Objective: To compare oocyte maturation, fertilization and cleavage rates, and embryonic developmental quality after culture of human immature oocytes from polycystic ovary syndrome (PCOS) patients in human tubal fluid (HTF) or tissue culture medium (TCM) 199. Design: Prospective, randomized, controlled trial. Setting: University hospital. Patient(s): Thirteen women undergoing 23 in vitro maturation cycles, from whom 119 oocytes were retrieved. Intervention(s): Cumulus-enclosed germinal vesicle-stage oocytes matured in TCM-199-supplemented or HTF-supplemented media. Main Outcome Measure(s): Oocyte maturation and fertilization rates, embryonic developmental quality. Result(s): Significant differences were observed between TCM 199 and HTF regarding maturation rate (82% vs. 56.9%), fertilization rate (70% vs. 39.4%), and embryo quality (81.3% vs. 41.7%). Conclusion(S): Human tubal fluid medium, although widely used for embryo fertilization and maintenance in IVF techniques, is not,in appropriate medium for the maturation of oocytes obtained from PCOS patients in nonstimulated cycles. (Fertil Steril(R) 2009;91:509-13. (C)2009 by American Society for Reproductive Medicine.)

Tracking the Ovarian Cycle in Black-and-Gold Howlers (Alouatta caraya) by Measuring Fecal Steroids and Observing Vaginal Bleeding

A better understanding of a species` reproductive physiology can help conservation programs to manage primates in the wild and develop assisted reproductive technologies in captivity. We investigated whether measurements of fecal progestin and estrogen metabolites obtained by a radioimmunoassay could be used to monitor the ovarian cycle of Alouatta caraya. We also compared the occurrence of vaginal bleeding with the hormone profiles. We collected fecal samples from 3 adult and 1 subadult captive female over 5 mo and performed vaginal cytology for the adults. The interval between fecal progestin surges in the adult females was 19.11 +/- 2.14 d (n = 18 cycles). Fecal progestin concentrations remained at basal values for 9.83 +/- 2.21 d (n = 18) and rose to elevated values for 9.47 +/- 0.72 d (n = 19). The subadult female showed basal levels of fecal estrogen and progestin concentrations throughout the study, suggesting that our hormone measurements are valid to monitor the ovarian cycle. Bleeding periods coincided with basal levels of fecal estrogens and progestin at intervals of 19.8 +/- 0.9 d and lasted for 4.1 +/- 1.0 d. Although we obtained these data from only 3 individuals, the results indicate that this species likely has a menstrual-type ovarian cycle. These data provide the first endocrine profile for the Alouatta caraya ovarian cycle and are similar to results obtained for other howler species. This similarity is important for comparative studies of howlers, allowing for a better understanding of their reproductive physiology and contributing to a critical information base for managing Alouatta species.

Birth of Koalas Phascolarctos cinereus at Lone Pine Koala Sanctuary following artificial insemination

This paper documents the successful development of an artificial insemination (AI) programme for the Koala Phascolurctos cinereus. The protocols for trials involving two methods to induce ovulation and two insemination techniques are described. In Trial 1, interrupted coitus using a 'teaser'♂ successfully induced ovulation in nine Koalas. Five ♀♀ were inseminated while conscious using a modified 'foley catheter' (Cook insemination catheter) resulting in the births of two offspring. The other four ♀♀ were anaesthetized and inseminated using a technique which allowed visualization of the most cranial portion of the urogenital sinus, where semen was deposited using a 3.5 Fr. 'Tom-cat catheter' (urogen-itoscopic insemination). Three of the four ♀♀ inseminated by this technique produced pouch young. Microsatellite analysis of DNA from the pouch young excluded the teaser ♀♀ as possible sires, confirming that all offspring were sired by donor sperm. In Trial 2, eight ♀♀ were induced to ovulate by injecting them with 250 International Units of human chorionic gonadotrophin (hCG). A luteal phase was confirmed in all eight ♀♀ but only one gave birth following urogenitoscopic insemination. The Koala pouch young in this study are the first of any marsupial to be conceived and born following A1 procedures. Details of the A1 procedures used are presented and the significance of A1 to the conservation biology of P. cinereus discussed.

As tecnologias de reprodução assistida e as representações sociais de filho biológico

Foram investigadas as representações sociais de filho biológico em casais que se submeteram às tecnologias reprodutivas. Nossos objetivos foram analisar a trajetória de vida do casal a partir do momento em que foi constatada a infertilidade e identificar as representações sociais de filho biológico. Cinco casais da Grande Vitória, ES foram entrevistados e nenhum deles havia obtido êxito com relação à gravidez após o tratamento. As entrevistas semi-estruturadas foram realizadas individualmente. O roteiro seguiu o mesmo padrão para ambos os cônjuges, incluindo itens sobre a constatação da infertilidade, o tratamento realizado, o significado da maternidade, paternidade, casamento e filho biológico. A análise dos resultados apontou para um fortalecimento do vínculo no casamento após o diagnóstico de infertilidade e evidenciou a importância que os casais atribuem ao filho biológico. Os elementos de representações sociais que apareceram fortemente foram: "sangue do meu sangue", descendência, semelhança física e pressão social.

An ecosystem of products and systems for ambient intelligence – the AAL4ALL users perspective

Developed societies are currently facing severe demographic changes: the world is getting older at an unprecedented rate. In 2000, about 420 million people, or approximately 7 percent of the world population, were aged 65 or older. By 2050, that number will be nearly 1.5 billion people, about 16 percent of the world population. This demographic trend will be also followed by an increase of people with physical limitations. New challenges will be raised to the traditional health care systems, not only in Portugal, but also in all other European states. There is an urgent need to find solutions that allow extending the time people can live in their preferred environment by increasing their autonomy, self-confidence and mobility. AAL4ALL presents an idea for an answer through the development of an ecosystem of products and services for Ambient Assisted Living (AAL) associated to a business model and validated through large scale trial. This paper presents the results of the first survey developed within the AAL4ALL project: the users’ survey targeted at the Portuguese seniors and pre-seniors. This paper is, thus, about the lives of the Portuguese population aged 50 and over.

Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing

INTRODUCTION: Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests. METHODS: After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen. RESULTS: In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions. CONCLUSIONS: Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

Le mariage à l'épreuve: les défis du partenariat de même sexe et de la procréation médicalement assistée en Suisse

Afin de rendre compte des luttes de?finitionnelles qui visent a? transformer ou a? pre?server le sens du mariage en tant qu?institution de l?he?te?rosexualite?, cet article examine deux de?bats cle?s qui ont re?cemment mis a? l?e?preuve l?institution du mariage en Suisse : l?institutionnalisation juridique des couples de me?me sexe avec l?adoption de la loi fe?de?rale sur le partenariat et la pre?servation du sche?ma familial he?te?rosexuel avec la loi fe?de?rale sur la procre?ation me?dicalement assiste?e. Ce texte de?fend que pour saisir l?e?volution du sens politique accorde? au mariage et a? la famille dans les de?bats sur le partenariat homosexuel, il convient d?e?tudier le processus d?institutionnalisation des couples homosexuels dans une temporalite? longue prenant place sur diffe?rentes sce?nes le?gislatives. This article examines how the heteronormative norm embedded in the institution of marriage has been put into question through the legal recognition of same sex couples and the access to assisted reproductive technologies in Switzerland. The paper shows that assessing the transformation of the meaning scheme attributed to the marriage and the family in the ongoing political debates on same-sex couples is enhanced by a diachronic analytical perspective focused on the different regulatory scenes, where the debate has taken place.

Trends in the prevalence, risk and pregnancy outcome of multiple births with congenital anomaly: a registry-based study in 14 European countries 1984-2007.

OBJECTIVE: To assess the public health consequences of the rise in multiple births with respect to congenital anomalies. DESIGN: Descriptive epidemiological analysis of data from population-based congenital anomaly registries. SETTING: Fourteen European countries. POPULATION: A total of 5.4 million births 1984-2007, of which 3% were multiple births. METHODS: Cases of congenital anomaly included live births, fetal deaths from 20 weeks of gestation and terminations of pregnancy for fetal anomaly. MAIN OUTCOME MEASURES: Prevalence rates per 10,000 births and relative risk of congenital anomaly in multiple versus singleton births (1984-2007); proportion prenatally diagnosed, proportion by pregnancy outcome (2000-07). Proportion of pairs where both co-twins were cases. RESULTS: Prevalence of congenital anomalies from multiple births increased from 5.9 (1984-87) to 10.7 per 10,000 births (2004-07). Relative risk of nonchromosomal anomaly in multiple births was 1.35 (95% CI 1.31-1.39), increasing over time, and of chromosomal anomalies was 0.72 (95% CI 0.65-0.80), decreasing over time. In 11.4% of affected twin pairs both babies had congenital anomalies (2000-07). The prenatal diagnosis rate was similar for multiple and singleton pregnancies. Cases from multiple pregnancies were less likely to be terminations of pregnancy for fetal anomaly, odds ratio 0.41 (95% CI 0.35-0.48) and more likely to be stillbirths and neonatal deaths. CONCLUSIONS: The increase in babies who are both from a multiple pregnancy and affected by a congenital anomaly has implications for prenatal and postnatal service provision. The contribution of assisted reproductive technologies to the increase in risk needs further research. The deficit of chromosomal anomalies among multiple births has relevance for prenatal risk counselling.

"El elemento volitivo como determinante de la filiación derivada de las técnicas de reproducción asistida"

Los cambio provocados por las Técnicas de Reproducción Asistida en materia de filiación nos llevan a pensar que los criterios decisivos para la disciplina de la filiación (por naturaleza) han cambiado a compás de las mutaciones de la conciencia social. En este trabajo procuraré demostrar cómo la voluntad ha pasado a ser el elemento decisivo en la determinación de la maternidad y/o paternidad a partir del surgimiento y utilización de las Técnicas de Reproducción Asistida.

Documento sobre congelación de ovocitos para la reproducción humana

Con este documento se pretende informar sobre la congelación de ovocitos para la reproducción humana y abordar el problema desde distintos punto de vista, con el fin de proporcionar argumentos y participar en el debate generado sobre el uso y aplicación de las técnicas de reproducción asistida y fomentar la necesaria actualización de una normativa que, si bien fue pionera en su momento, presenta hoy las carencias y contradicciones que el avance científico y el devenir social han ocasionado, y que requieren su puesta al día.

Documento sobre selección de sexo

El grup ha analitzat la possibilitat de seleccionar el sexe dels futurs fills, qüestió que avui dia suscita controvèrsia ja que la prohibició legal generalitzada que actualment hi ha s¿està posant en dubte.

Reproducción humana asistida: los problemas que suscita desde la bioética y el derecho

El artículo trata de los problemas éticos y jurídicos que surgen a raíz de la puesta en práctica de técnicas de reproducción asistida, al querer determinar la paternidad o poder elegir el sexo de un embrión viable, entre otros.

Documento sobre selección de sexo

El grup ha analitzat la possibilitat de seleccionar el sexe dels futurs fills, qüestió que avui dia suscita controvèrsia ja que la prohibició legal generalitzada que actualment hi ha s¿està posant en dubte.