Justificativa: As técnicas de reprodução assistida (TRA), usadas para o tratamento de infertilidade/subfertilidade, incluem manipulação in vitro de oócitos e esperma, ou embriões, com o objetivo de alcançar gravidez e nascimentos vivos. O recrutamento de múltiplos folículos é fundamental para o aumento das taxas de gravidez, e isso é alcançado por meio do estímulo ovariano controlado (EOC). A monitorização do EOC é realizada por contagem de folículos ovarianos e medidas ultrassonográficas, associadas ou não à dosagem hormonal. Justificase monitorar a fase folicular para decisões a respeito da dose de gonadotrofinas administradas, detecção do risco de ocorrência da síndrome de hiperestímulo ovariano (SHO) e do planejamento do triggering da maturação final; porém a necessidade da monitorização intensiva da EOC é controversa, pois a combinação dos métodos consome mais tempo, recursos e está associada com maior desconforto para a paciente. Objetivo: Avaliar a eficácia e segurança da monitorização da EOC em ciclos de reprodução assistida, usando somente ultrassonografia (US). Métodos de busca: As buscas por estudos randomizados foram realizadas nos principais bancos de dados eletrônicos. Além disso, foram examinadas, manualmente, as listas de referências dos estudos incluídos em revisões semelhantes. A última busca eletrônica foi realizada em 12 de março de 2015. Critérios de Seleção: Apenas estudos verdadeiramente randomizados, que comparassem a monitorização do EOC por US associado à dosagem hormonal e US isoladamente, monitorização do EOC por US2D e US3D, bem como US2D e telemonitorização endovaginal operada pela própria paciente (SOET), foram considerados elegíveis. Os estudos que permitiam a inclusão de uma mesma paciente duas vezes foram incluídos apenas se os dados do primeiro ciclo estivessem disponíveis. Coleta e Análise de Dados: Dois revisores avaliaram, independentemente, a elegibilidade, extração de dados e os riscos de viéses dos estudos incluídos. Quaisquer discordâncias foram resolvidas em consulta com um terceiro revisor. Quando necessário, os autores dos estudos incluídos foram contatados para maiores informações. Resultados: Foram selecionados 1717 registros, 10 dos quais eram elegíveis. Nenhum estudo relatou nascidos vivos. Seis estudos compararam a monitorização do EOC por US isolada com US associada à dosagem hormonal. Os intervalos de confiança (IC) foram extensos e não permitiram concluir a existência de benefício nem prejuízo associado ao uso de US isolada, em relação aos desfechos SHO (odds ratio - OR=1.03, IC95% 0.48 a 2.18, p=0.95) e abortamento (risco relativo - RR=0.37, IC95% 0.07 a 1.79, p=0.21). Para gravidez clínica, o IC foi compatível com pequeno benefício a pequeno prejuízo (RR=0.96, IC95% 0.80 a 1.16, p=0.70). Para número de oócitos captados, o IC foi compatível com apreciável benefício a não efeito (Diferença média MD=0.92 oócitos captados, CI95% -0.19 a 2.04, p=0.70). Dois estudos compararam US3D e US2D e os IC foram extensos e não permitiram concluir pela existência de benefício nem prejuízo associado à monitorização por US3D para os desfechos: gravidez clínica (RR=1.00, IC95% 0.58 a 1.73) e número de oócitos captados (MD= -0.37 oócitos, IC95% -3.63 a Resumo 2.89). Apenas um estudo comparou monitorização por US2D convencional com SOET, e o IC observado foi amplo e não permitiu concluir pela existência de benefício nem prejuízo associado à SOET, considerando se gravidez clínica (RR=0.95, IC 95% 0.52 a 1.75) e número de oócitos captados (MD=0.50, CI 95% - 2.13 a 3.13). Conclusão: No que concerne à eficácia, as evidências atuais sugerem que monitorizar o EOC apenas com US não deva alterar, substancialmente, as chances de se alcançar gravidez clínica. O número de oócitos captados é similar ao se comparar com a monitorização por US associada à dosagem hormonal. Quanto à segurança, também não houve aumento no risco de desenvolvimento de SHO. Contudo, a interpretação dos resultados deve ser realizada com cautela, já que para todos os desfechos e todas as comparações, os dados disponíveis são inconclusivos, pois a qualidade de evidência foi comprometida por imprecisão e falha dos estudos em relatar a metodologia aplicada. Por isso acredita-se que serão necessários mais estudos avaliando o procedimento ideal para monitorização da EOC / Background: The assisted reproductive techniques (ART) for the treatment of infertility/subfertility, include in vitro handling of both human oocytes and sperm or of embryos with the objective of achieving pregnancy and live birth. The recruitment of multiple follicles is often necessary for better results in pregnancy rates and it\'s achieved by performing controlled ovarian stimulation (COS). COS monitoring is performed by ovarian follicle counting and ultrasonography measurements and / or hormones dosage. It is appropriate to monitor the follicular phase for decisions regarding administered of gonadotropin dose, to assess the risk of ovarian hyperstimulation syndrome (OHSS), to determine the best time to trigger final follicular maturation. However, the need for intensive COS monitoring is controversial: the combination of the methods adds costs and discomfort for the woman who is undergoing ART and requires additional time. Objectives: To evaluate the efficacy and safety of monitoring controlled ovarian stimulation by ultrasound in assisted reproduced tecniques. Search Methods: The searches for randomized controlled trials (RCT) were performed in the main electronic databases; in addition, we hand searched the reference lists of included studies and similar reviews. We performed the last electronic search on March 29, 2015. Selection Criteria: Only truly randomized controlled trials comparing COS monitoring by ultrasonography and/or hormonal assessment, as studies comparing COS monitoring by 2DUS and 3DUS were considered eligible. We included studies that permitted the inclusion of the same participant more than once (cross-over or \'per cycle\' trials) only if data regarding the first treatment of each participant were available. Data Collection and Analysis: Two reviewers independently performed study eligibility, data extraction, and assessment of the risk of bias and we solved disagreements by consulting a third reviewer. We corresponded with study investigators in order to resolve any queries, as required. Results: The search retrieved 1717 records; ten studies were eligible. No study reported live birth. Six studies compared US only vs. US + Hormones. The confidence intervals (CI) were large and did not allow us conclude benefit or harm associated with the US Only for both OHSS (Odds ratio - OR=1.03, 95%CI 0.48 to 2.18, P=0.95), and miscarriage (relative risk - RR=0.37, 95%CI 0.07 to 1.79, P=0.21). For clinical pregnancy, the CI was compatible with small benefit to small harm (RR=0.96, 95%CI 0.80 to 1.16, P=0.70). For the number of oocytes retrieved, the CI was compatible with appreciable benefit to no effect (Mean difference - MD=0.92 oocytes, 95%CI -0.19 to 2.04, P=0.70). Two studies compared 3DUS vs. 2DUS: the confidence intervals (CI) were large and did not allow us conclude benefit or harm associated with 3DUS regarding clinical pregnancy (RR=1.00, CI95% 0.58 to 1.73) and number of oocytes retrieved (MD= -0.37 oocytes, 95%CI -3.63 to 2.89). One study compared 2DUS vs. SOET, the CI was large and did not allow us conclude benefit or harm associated with SOET regarding clinical pregnancy (RR=0.95, 95%CI 0.52 a 1.75) and number of oocytes retrieved (MD=0.50, 95%CI -2.13 a 3.13). Authors\' Conclusions: Regarding effectiveness, current evidence suggests that monitoring COS only by US only should not change substantially the chances of achieving clinical pregnancy. The number of retrieved oocytes is similar to compare with the monitoring by US associated with hormonal assessment. security also seems not to increase the risk of developing OHSS. However the interpretation of results should be performed with caution, since for all outcomes and comparisons, the available data are inconclusive because the quality of evidence was compromised by inaccuracy and poor reporting of study methodology. So we believe that further studies evaluating the ideal procedure for monitoring the COS are needed

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No. of bitstreams: 1 kallinecarvalhogoncalveseler.pdf: 1185811 bytes, checksum: 4ba1b7ea9d9aae62dffb895f2c49c11a (MD5) Previous issue date: 2015-05-08 / O presente trabalho busca uma melhor interpretação das novas tecnologias reprodutivas contrapondo-se ao viés cientificista dos projetos de lei em tramitação no país que primam pela defesa dos interesses dos profissionais, em especial clínicas e laboratórios, desconsiderando os direitos do novo ser gerado. A relevância social e científica do tema está em se refletir acerca da necessidade de atenção e cautela no implemento das novas tecnologias destinadas à reprodução. Hoje, o avanço tecnológico está intimamente vinculado aos meios de aquisição de poder e carece de construções valorativas. Isso justifica a necessidade crescente de um maior fortalecimento da proteção jurídica do embrião extracorporal a fim de que o Princípio da Dignidade da Pessoa Humana seja efetivamente concretizado. À luz da Constituição, a pessoa humana não é categorizada como sujeito que contrata, que constitui formalmente uma família e que tem um patrimônio. A proteção constitucional é dirigida à dignidade da pessoa, considerada em todas as suas emanações. O objetivo precípuo deste trabalho está em buscar um enquadramento ético-jurídico para o embrião oriundo das técnicas de reprodução assistida com o intuito de sustentar seu status pessoal. Para persecução deste fim, adota-se como metodologia a análise de conteúdo, tomando-se por marco teórico o conceito de consciência presente na fenomenologia de Husserl e as ideias semelhantes do grupo denominado substancialista que sustenta um conceito onto-axiológico de pessoa. Em um segundo momento, a partir do pressuposto de que o embrião extra corporal é pessoa e, portanto, sujeito de direitos personalíssimos, intenta-se demonstrar a inconstitucionalidade da produção dos embriões excedentários e propor a substituição dessa prática pela técnica vitrificação de ovócitos – técnica promissora para o tratamento de infertilidade da reprodução assistida. A pesquisa proposta alinha-se, assim, à vertente das pesquisas jurídico-compreensivas e jurídico-propositivas. / This paper pursues a better interpretation of the new reproductive technologies by making a counterpoint to the scientistic bias of bills processing in the country which excels in defending the interests of professionals, especially in clinics and laboratories, disregarding the new person generated. The social and scientific relevance of the subject is to reflect on the necessity of attention and caution in the implementation of the new technologies for breeding. Today, technological advancement is closely tied to the means of acquiring power and lacks evaluative constructs. It justifies the growing need for a further strengthening of the legal protection of the extracorporeal embryo so that the Principle of Human Dignity is effectively implemented. In Constitution’s perspective, the human person is not categorized as a subject who hires, who formally constitutes a family and who has a heritage. The constitutional protection is directed to the dignity of the person, considered in all its emanations. The primary objective of this research is to seek an ethical-legal framework for the embryo arising from assisted reproduction techniques in order to sustain the personal status. To attain this end, it will be adopted the content analysis methodology, taking as theoretical framework the consciousness in Husserl phenomenological meaning and the similar ideas of the group called substantialistic which sustain an onto-axiological concept of person. In a second moment, starting from the assumption that the extra-corporeal embryo is a person and, therefore, subject of personal rights; intends to demonstrate the unconstitutionality of the production of surplus embryos and to propose the replace of this practice by the cryopreservation of oocytes by vitrification - a promising new technique for assisted human reproduction. The legal proposed research is both comprehensive and purposeful.

<p>Infertility is the disease of the reproductive system and is estimated to affect more than 10% of the people of reproductive age. Assisted reproductive technologies (ART) are methods designed to alleviate infertility problems. <i>In vitro </i>embryo production is part of most infertility treatments and the efficiency of ART is low due to the lack of reliable methods to measure embryo viability. In order to improve the success rate of ART procedures, the current study was designed to investigate the use of an optical analyzer technology known as the Biodynamic Imaging (BDI) system for viability assessment. BDI is a novel approach that is able to measure intracellular dynamic processes that are directly related to functional events. During a series of experiments, 13 different biomarkers of oocytes and embryos were monitored by the BDI microscope and used for machine learning and evaluation of BDI sensitivity. We monitored cellular mechanisms essential for proper embryo development such as (1) extrusion of first and second polar body (2) energy status and mitochondrial activity, and (3) viability of embryos with different cellular composition. We were able to identify several biomarkers that have the potential to indicate viability: (1) slope, (2) NSD, (3) Knee (4) Floor, and (5) R² could consistently differentiate between oocytes and embryos of different viability. In addition, the BDI microscope could successfully predict the energy status of embryos by identifying 4 biomarkers (Slope, Knee, Floor, and Dy). Finally, a lipidomic analysis was done to evaluate the lipid composition of oocytes with different cytoplasm integrities. This analysis demonstrated that there is a difference in lipid subclasses among oocytes with dark vs. light cytoplasm. The results indicate that the BDI is useful for viability assessment of oocytes and embryos and may be helpful for the improvement of the efficiency of assisted reproductive technologies.</p>

This work "Is assisted reproduction a therapy for infertility?" deals with methods of assisted reproduction. The aim is not only to elucidate its techniques but also to open some difficult ethical questions. This work shows why we cannot regard assisted reproduction as a legitimite treatment for infertility and suggests alternative procedures. The first chapter is about the individual human life seen from the perspective of various sciences. The second chapter is about causes of infertility and about possibilities of its treatment. The third chapter deals with various methods of artificial reproduction. The following chapter is about ethical challenges related to these methods. The fifth chapter presents the standpoint of the magisterium of the Catholic Church. The final chapter tries to see fertility and parenthood from a larger perspective: the principal alternatives are substitute family care and spiritual parenthood. Keywords methods of assisted reproduction, infertility, parenthood, substitute family care, spiritual fertility

The main subject of this master thesis is the issue of surrogacy in the Czech Republic and the Federal Republic of Germany. At the beginning, this thesis aspires to define the term surrogacy in general as well as other connected terminology, and subsequently to specify its categories and describe its major historic milestones. Afterwards, it deals with the rather brief Czech legislation concerning this institute and the consequences this has inevitably led to. It also concentrates on the methods of the assisted reproduction which help to put the surrogate motherhood into practice. This thesis does include ethical problems of surrogacy and some of the psychological and sociological aspects as well. Consequently, it summarizes sanctions which may arose as a result of surrogacy. After that, it explains the legislation related to this issue in the Federal Republic of Germany, its history and legal limits. It deals with the German sanctions which the realization as well as mere arrangement may be subjected to. It examines the standpoint of the German legislator regarding the right of a child to know his or her origin including its consequences, such as non-anonymous sperm donation and obligation of the legal parents to undergo a DNA test. It describes most recent demands of the society regarding the new...

The diploma thesis conceptualizes an experience of women with assisted reproduction and services offered in fertility clinics. The assisted reproduction is a medical method treating infertility. The scientific progress and the usage of new reproductive technologies bring up broad possibilities in reproduction. Regarding the biological explanation of human's life, the biomedicine discourse represents a dominant approach in reproduction and claims control over a conception of a new life. Women undergoing the assisted reproduction, which is offered in case of infertility, face new challenges related to social, economic, ethical, and legal issues. The major goal of this thesis is the analysis of an experience of women with assisted reproduction in state-funded and private fertility clinics while applying a feminist critical reading. Qualitative research is based on semi-structured interviews with women who have turned to medical experts for help and underwent the assisted reproduction or other reproductive methods to increase a chance of conception. Key words: assisted reproduction, reproduction, women's experience, reproductive medicine, fertility, gender, poststructuralism

This diploma thesis aims to describe policy design of assisted human reproduction in the Czech Republic in the frame of two examined groups: 1) women undergoing this procedure, 2) service providers. The goals are also to identify the policy framework of assisted human reproduction in the political environment. The theory background is supported by the theory of social construction of the target audience written by the authors Ingram and Schneider and the analysis of the framework by Rein and Schon. The legislation of assisted human reproduction is covered mostly in the period of 2011 - 2021. The analytical part of this work is based on systematic expert surveys, questionnaire surveys and analysis of stenographic records. The benefits and drawbacks arising from the legislations, political power and social constructions in relation to both examined groups are formulated in the paper's results. Furthermore, two competing policy frameworks are identified during the debate of amendments to the Act on Specific health Services and the Act on Public Health Insurance.

Η ενδοκυττάρια έγχυση σπέρματος (ICSI) εισήχθη ως μέθοδος υποβοηθούμενης αναπαραγωγής (ΑRT) κυρίως για την αντιμετώπιση της ανδρικής στειρότητας. Όμως, λόγω των υψηλών ποσοστών επιτυχίας, και παρά τις αυξανόμενες ανησυχίες για τους κινδύνους από τη σημαντική ανθρώπινη παρεμβολή στο γονιδίωμα, το επιγονιδίωμα και την ανάπτυξη των παιδιών, προτιμάται ακόμα και σε περιπτώσεις όπου δεν απαιτείται ιατρικά. Από τις λίγες σήμερα συστηματικές μελέτες παιδιών από ART, καταγράφεται αυξημένο ποσοστό προδιάθεσης τους σε ασθένειες που σχετίζονται με κακό καρδιομεταβολικό πρότυπο στην ενήλικη ζωή. Στόχος της εργασίας ήταν η διερεύνηση της δυνατότητας χρήσης της μεταβολομικής ανάλυσης για τον πρώϊμο και έγκυρο προσδιορισμό σχετικών διαταραχών σε δείγματα πλάσματος προεφηβικών κοριτσιών και αγοριών από ΙCSI, που επιλέχτηκαν από ένα συστηματικά χαρακτηρισμένο σύνολο παιδιών μελέτης της Α’ Παιδιατρικής Κλινικής, Νοσοκομείου «Αγία Σοφία», Ιατρικής Σχολής, ΕΚΠΑ. ΥΛΙΚΑ ΚΑΙ ΜΕΘΟΔΟΙ: Τα μεταβολικά πρότυπα πλάσματος (α) 10 κοριτσιών από ΙCSI και 10 από φυσιολογική γονιμοποίηση (NC) και (β) 16 αγοριών από ΙCSI και 16 από NC ποσοτικοποιήθηκαν με χρωματογραφία αερίων – φασματομετρία μάζας (GC-MS). Μετά από την ταυτοποίηση κορυφών και την κατάλληλη κανονικοποίηση των προτύπων, 86 πρότυπα 70 μεταβολιτών στα κορίτσια και 92 πρότυπα 80 μεταβολιτών στα αγόρια αναλύθηκαν ξεχωριστά, και συγκριτικά με αλγορίθμους πολυπαραμετρικής στατιστικής ανάλυσης των λογισμικών TM4-MeV (v.4.9.0), και ΧLSTAT (v.2013.4.03). Οι διαφορές στο πρότυπο σύστασης του πλάσματος σε μικρού μεγέθους μεταβολίτες μεταξύ των ΙCSI και ΝC ομάδων σε κορίτσια και αγόρια και μεταξύ των δύο φύλων οπτικοποιήθηκαν σε κατάλληλα ανακατασκευασμένο από τη βιβλιογραφία και σχετικές βάσεις δεδομένων μεταβολικό δίκτυο πολλών ιστών. ΑΠΟΤΕΛΕΣΜΑΤΑ: Στα κορίτσια, ο αλγόριθμος μερικών ελαχίστων τετραγώνων-διακριτής ανάλυσης (PLS-DA) κατέδειξε σαφή διαχωρισμό των μεταβολικών πρoτύπων μεταξύ των ομάδων ΙCSI και NC. Ο διαχωρισμός αυξάνεται με το συνυπολογισμό των βιοχημικών μετρήσεων. Στα αγόρια, η PLS-DA των μεταβολικών ή και βιοχημικών προτύπων κατέδειξε επίσης διαχωρισμό, αν και μικρότερο, σε σχέση με κορίτσια. Η ανάλυση σημαντικότητας για μικροσυστοιχίες (SAM), που ενδείκνυται για την ανάλυση ομικών δεδομένων, ανέδειξε 37 από τους 70 μεταβολίτες που αναλύθηκαν στα κορίτσια με σημαντικά διαφορετική συγκέντρωση μεταξύ των ΙCSI και ΝC ομάδων, με 34 από αυτούς να αυξάνονται στην ICSI ομάδα. Οι 34 μεταβολίτες αφορούν κύρια σε σάκχαρα, αλκοόλες και οξέα σακχάρων, οργανικά οξέα και λιπίδια, που έχουν συνδεθεί με αντίσταση στην ινσουλίνη, μεταβολικό σύνδρομο, ή/και την παχυσαρκία. Η ίδια ανάλυση στα αγόρια ανέδειξε 25 από τους 80 μεταβολίτες που αναλύθηκαν με χαρακτηριστική διαφορά μεταξύ των ομάδων ICSI και NC, εκ των οποίων 9 με σημαντικά μικρότερη συγκέντρωση στην ομάδα ΙCSI. Χαρακτηριστικά αναφέρεται ότι στους 9 μεταβολίτες ανήκουν οι 4 με την πλέον διαφορετική συγκέντρωση μεταξύ των ICSI και NC ομάδων, που είναι η σορβιτόλη, και τα αρωματικά αμινοξέα τρυπτοφάνη, φαινυλαλανίνη και τυροσίνη. Συγκριτική ανάλυση του μεταβολικού προτύπου των δύο φύλων στην NC ομάδα κατέδειξε μια σαφή διαφοροποίηση, η οποία φαίνεται να αποτελεί κύρια αιτία της παρατηρούμενης φυλο-ειδικής μεταβολικής διαφοροποίησης μεταξύ των ομάδων ICSI και ΝC. ΣΥΜΠΕΡΑΣΜΑΤΑ: Η πολυπαραμετρική ανάλυση της σύστασης του πλάσματος σε μικρού μοριακού βάρους μεταβολίτες επέτρεψε τον προσδιορισμό μεταβολικών διαφορών μεταξύ των ομάδων ICSI και NC, που υποστηρίζουν την προδιάθεση των παιδιών από ICSI σε αντίσταση στην ινσουλίνη, με διακριτούς όμως μεταβολικούς και βιοχημικούς δείκτες μεταξύ των δύο φύλων. Τα ευρήματα αυτά πρέπει να επιβεβαιωθούν σε ένα ευρύτερο σύνολο παιδιών και των δύο φύλων. Καταδεικνύουν όμως την αξία της μεταβολομικής να παρέχει μία υψηλής ευκρίνειας προοπτική της μεταβολικής κατάστασης, οδηγώντας στον προσδιορισμό χαρακτηριστικών μεταβολικών προτύπων ακόμα και σε πολύπλοκες καταστάσεις φυσιολογίας. / The intracytoplasmic sperm injection (ICSI) method was introduced in artificial reproduction technology (ART) mainly to treat male infertility. However, due to its high success rates and despite the growing concerns concerning the risk that the significant human intervention associated with this method may have to the genome, epigenome and development of the offspring, the use of ICSI has gradually increased in the recent years, even when it is not medically required. Based on the few currently available systematic studies of ART conceived children, the latter are considered of higher risk for cardio-metabolic diseases as adults. The goal of the present study is to investigate whether metabolomic analysis of the blood plasma could contribute to the early and accurate determination of relevant predisposition in ICSI conceived prepubertal girls and boys, specifically selected from a systematically characterized group of children, participated in a study of the First Department of Pediatrics of the “Agia Sophia” Hospital, Medical School, University of Athens. MATERIALS AND METHODS: The blood plasma metabolic profiles of (a) 10 ICSI- and 10 naturally conceived (NC) girls and (b) 16 ICSI and 16 NC boys were acquired using gas chromatography-mass spectrometry. After peak identification and appropriate normalization, 86 profiles of 70 metabolites in girls and 92 profiles of 80 metabolites in boys were analyzed separately and comparatively using multivariate statistical analysis algorithms of TM4-MeV (v.4.9.0) and XLSTAT (v.2013.4.03) software. The differences in the plasma metabolite concentration profiles between the ICSI and NC groups in girls and boys were visualized in an inter-tissue metabolic network that was reconstructed based on relevant literature and metabolic databases. RESULTS: For the girls, the algorithm of partial least squares-discriminant analysis (PLS-DA) indicated a clear differentiation of the metabolic profiles between the ICSI and NC groups. The discrimination is more pronounced, when biochemical data are also considered. For the boys also, PLS-DA indicated separation between the metabolomic profiles of the two groups analyzed individually or in combination with the biochemical data, but not as explicit as in girls. Significance analysis for microarrays (SAM) determined 37 out of the 70 analyzed metabolites in the plasma profiles of the girls with significantly different concentration between the ICSI and the NC groups; 34 of these were of higher concentration in the ICSI group. The 34 metabolites include mainly sugars, sugar alcohols and acids, organic acids and lipids that have been associated with insulin resistance, metabolic syndrome and/or obesity. The same analysis in the plasma profiles of the boys determined 25 out of the 80 analyzed metabolites with significant difference between the ICSI and NC groups; nine of these were of significantly lower concentration in the ICSI group. It is underlined that the four most discriminatory metabolites between the ICSI and NC groups, i.e. sorbitol and the aromatic amino acids tryptophan, phenylalanine and tyrosine, are among the nine negatively significant. Comparative analysis of the metabolic profiles between the two sexes within the NC group indicated an unequivocal differentiation, which is considered to be the main cause of the observed sex-specific metabolic differences between the ICSI and NC groups. CONCLUSIONS: The multivariate statistical analysis of blood plasma metabolite profiles enabled the determination of sex-specific metabolic differences between the ICSI and NC groups; these differences support increased predisposition to insulin resistance for the ICSI offspring, with clearly different, however, metabolic and biochemical markers in the two sexes. These findings need to be confirmed in a wider group of children. They demonstrate, however, the value of metabolomics to provide a high-resolution perspective of the metabolic state, leading to the determination of characteristic metabolic profiles even in complex physiological conditions.

Les marqueurs épigénétiques, en particulier la méthylation de l’ADN des gènes soumis à empreinte parentale, sont sensibles aux changements environnementaux. Les techniques de l’aide médicalisée à la procréation (AMP) nécessitant la manipulation des gamètes et des embryons in vitro et dans la plupart des cas la stimulation hormonale de l’ovulation des patientes, peuvent interférer avec la reprogrammation et/ou le maintien de la méthylation des gènes soumis à empreinte. Afin d’évaluer ce risque nous avons analysé le profil de méthylation de KvDMR1, qui régule l’expression de KCNQ1OT1, dans des ovocytes humains mûris in vivo ou in vitro, provenant de patientes stimulées ou non. Nos résultats montrent que la mise en place de la méthylation au niveau de KvDMR1 se poursuit au cours de la maturation de l’ovocyte après reprise de la méiose, in vivo et in vitro et que la superovulation des patientes en AMP génère des ovocytes épigénétiquement immatures. Par ailleurs, l’étude de la méthylation de KvDMR1 et de H19 DMR (qui régule l’expression d’Igf2 et H19) dans des embryons issus d’ICSI, évolutifs ou présentant un défaut de développement, n’établit pas de lien entre les dérégulations de l’empreinte et l’arrêt du développement embryonnaire au stade blastocyste. / Epigenetic modifications, particularly DNA methylation of imprinted genes are sensible to environment. Techniques of assisted reproduction require in vitro manipulation of gamete and embryos and currently superovulation of patients. These technologies may interfere with eprogramming and maintenance of methylation at imprinted genes. To evaluate such a risk, we have determined the methylation profile of KvDMR1, the region that regulates KCNQ1OT1 imprinted gene, in human oocytes retrieved from stimulated or unstimulated cycles, at different phases of their maturation in vivo or in vitro. Our results show that the timing of establishment of the methylation profile of KvDMR1 covers the maturation phase of 199 oocyte growth, in vivo and in vitro, and that hyperstimulation likely recruits young follicles epigenetically immature. Analysis of the methylation profile of KvDMR1 and H19DMR (DMR of IGF2/H19) in ICSI embryos suggests that imprinting disorders are not responsible of embryo developmental failure prior the blastocyst stage

<p>Uvod: Vi&scaron;eplodne trudnoće se javljaju u 1,5% svih trudnoća nakon spontane koncepcije, dok nakon postupaka vantelesne oplodnje ovaj postotak u Evropi iznosi preko 20% uz velike varijacije među zemljama. U na&scaron;oj sredini, stopa vi&scaron;eplodnih trudnoća nakon postupaka vantelesne oplodnje iznosi daleko iznad 30%. Pojava hipertenzivnog sindroma u trudnoći, gestacijskog dijabetesa, operativnog zavr&scaron;avanja trudnoće, prevremenog porođaja, male porođajne telesne mase, neurolo&scaron;kih sekvela kod rođene dece i gotovo svih drugih komplikacija po majku i plod, kao i celokupno opterećenje zdravstvenog sistema vi&scaron;estruko su veći kod vi&scaron;eplodnih u odnosu na jednoplodne trudnoće i udeo navednih komplikacija raste sa brojem plodova. Sa druge strane deca iz postupaka vantelesne oplodnje čine i do 4,5% sve živorođene dece u pojedinim zemljama, &scaron;to uz činjenicu da infertilitet pogađa 16-18% parova u na&scaron;oj sredini daje ovoj pojavi posebnu dimenziju i činije i dru&scaron;tvenim problemom. Perinatalni ishodi trudnoća iz postupaka vantelesne oplodnje su u velikoj meri kompromitovani visokom stopom multiplih trudnoća, koje se danas smatraju komplikacijom, a ne uspehom postupaka vantelesne oplodnje. Jednoplodne trudnoće iz postupaka vantelesne oplodnje u većim studijama pokazuju diskretno slabije perinatalne ishode u odnosu na one spontano začete, dok kod vi&scaron;eplodnih trudnoća ova korelacija nije jasno izražena i dokumentovana, uz prisutnu dilemu da li je vi&scaron;eplodnost sama po sebi ili način koncepcije glavni problem u zapaženoj pojavi. Cilj rada: Uporediti perinatalne ishode vi&scaron;eplodnih trudnoća nastalih postupcima vantelesne oplodnje i spontano začetih kao i perinatalne ishode jednoplodnih i vi&scaron;eplodnih trudnoća iz postupaka vantelesne oplodnje. Pored navdenog cilj rada je i ukazati sveobuhvatnost navedenog problema i na moguća re&scaron;enja za smanjenje njihove učestalosti. Materijal i metode: Kombinacijom retrospektivne opservacione studije i prospektivne longitudinalne kohortne studije u periodu analizom perinatalnih ishoda pacijentkinja porođenih na Klinici za ginekologiju i aku&scaron;erstvo Kliničkog centra Vojvodine u periodu od od 01.01.2008. do 31.12.2010. godine, studija je analizirala i poredila perinatalne ishode kod 174 spontano začete vi&scaron;eplodne trudnoće, 163 vi&scaron;eplodne trudnoće nastale postupkom vantelesne oplodnje, kao i 155 jednoplodnih trudnoća začete postupkom vantelesne oplodnje. Analizirani parametric bili su telesna masa novorođenčeta, dostignuta gestacijska starost, vrednosti Apgar skora, učestalost hipertenzivnog sindroma kod majke i brojni drugi parametri perinatalnog ishoda. Uzeti od strane obučenih kliničara i uno&scaron;eni u posebno dizajniranu bazu podataka, rezultati su statistički analizirani u program JMP ver 9.0 (SAS publisher) uz kori&scaron;ćenje ANOVA analize za testiranje statističke značajnosti između srednjih vrednosti kontinuiranih varijabli, dok je statistička značajnost razlike učestalosti kategorijskih varijabli je određivana Pearsonovim &chi;2 testom. Rezultati: Jednoplodne ART trudnoće uz prosečnu starost od 33,5 godine, prosečnu gestacijsku starost na porođaju od 38,26 gn, udeo prevremenih porođaja od 12,9%, prosečnu telesnu masu od 3258 g, AS u prvom minutu od 8,35 i u petom minutu od 9,2, stopu carskog reza od 65,81%, udeo GDM-a od 7,1%, anemije od 41,94% i preeklampsije od 4,52%, ima sve relevantne parametre perinatalnog ishoda statistički značajno (p&lt;0.0001) superiornije od kako ART tako i non ART blizanačkih trudnoća. ART blizanačke trudnoće pokazale su prosečnu starost majke od 32,9 godina, prosečnu gestacijsku starost na porođaju od 35,6 gn, udeo prevremenih porođaja od 58,27%, prosečnu telesnu masu od 2374 g, AS u prvom minutu od 7,45 i u petom minutu od 8,65, stopu carskog reza od 83,7%, udeo GDM-a od 15,11%, anemije od 78,42% i preeklampsije od 12,23%, dok su non ART blizanačke trudnoće pokazale prosečnu starost majke od 28,8 godina, prosečnu gestacijsku starost na porođaju od 36,08 gn, udeo prevremenih porođaja od 49,71%, prosečnu telesnu masu od 2433 g, AS u prvom minutu od 7,75 i u petom minutu od 8,75, stopu carskog reza od 58,33%, udeo GDM-a od 7,02%, anemije od 67,84% i preeklampsije od 11,11%. Pored godina majke i udela carskog reza koji su bili vi&scaron;i u ART blizanačkim trudnoćama (&lt;0.0001), kao i blago veće pojavi poremećaja količine plodove vode (p=0,033), gotovo svi ostali pokazatelji toka i ishoda trudnoće bili su komparabilni u navedenim grupama. Diskusija i zaključak: Studija je pokazala da su tok i ishod vi&scaron;eplodnih trudnoća nastalih spontano i postupcima vantelesne oplodnje ekvivalentni u gotovo svim pokazateljima uz sličnu prosečnu telesnu masu i gestacijsku starost novorođenčadi, kao i da su svi navedeni parametri ovih vi&scaron;eplodnih trudnoća bez obzira na način koncepcije upadljivo i podjednako lo&scaron;iji u poređenju sa jednoplodnim trudnoćama iz postupka vantelesne oplodnje. Izuzimajući vi&scaron;eplodnost kao factor rizika deca iz postupaka vantelesne oplodnje su generalno zdrava. Sama vi&scaron;eplodnost, a ne način koncepcije predstavljaju problem, koje se sa pravom smatra najvećom komplikacijom vantelesne oplodnje. Dodatna analiza iskustava drugih zdravstvenih sistema ukazuje da jedino &scaron;iroka i sveobuhvatna implementacija strategije vraćanja samo jednog embriona (Single embryo transfer &ndash; SET) može da dovede do smanjivanje stope multiplih trudnoća nakon postupaka vantelesne oplodnje, i sledstvenih komplikacija, a bez ugrožavanja samog uspeha vantelesne oplodnje. Iskustva drugih zdravstvenih sistema ukazuju da je uspe&scaron;na implementacija SET-a jedino moguća uz angažovanje celog dru&scaron;tva, zajedno sa brojnim legislativnim merama iz domena nadzora, kontrole i finansiranja postupaka vantelesne oplodnje. Obim i način finansiranja postupaka vantelesne oplodnje od strane države (uz vi&scaron;e besplatnih poku&scaron;aja za infertilne parove) uz obaveznu upotrebu SET-a, i sistema krioprezervacije na osnovu primera iz prakse predstavlja ključ u borbi za smanjenje problema vi&scaron;eplodnih trudnoća nakon postupaka vantelesne oplodnje.</p> / <p>Introduction: Multiple pregnancies occur in 1.5% of all pregnancies after spontaneous conception and in more than 20 % of all pregnancies concieved after assisted reproductive technologies in Europe, with large variations between countries. In our setting, the rate of multiple pregnancies after the ART is well above 30%. The occurrence of hypertensive syndrome in pregnancy, gestational diabetes, operative delivery, premature birth, low birth weight, neurological and developmental impairment in children, and almost all the other complications for the mother and fetus, as well as the entire burden of the health system are several times higher in multiple pregnancies compared with singleton pregnancies. Incidence of&nbsp; forementioned complications rises with number of fetuses. On the other hand, children from in vitro fertilization procedures make up 4.5% of all live births in some countries, which together with the fact that infertility affects aproximately 16-18% of couples in our country gives an extra dimension to this phenomenon and makes it not just medical but wider social problem. Perinatal outcomes of pregnancies after assisted reproductive technologies (ART) are greatly compromised by the high rate of multiple pregnancies, which are now considered to be a complication rather than success of ART procedures. ART Singleton pregnancies have, in larger studies, show discretely lower perinatal outcomes compared with those conceived spontaneously, while for the multiple pregnancies, this correlation is not clearly expressed and documented. There remains dilemma whether multiplicity itself or the way of conception (ART vs. non ART) constitutes a major problem in the observed differences regarding perinatal outcome of ART pregnancies. Objective: To compare the perinatal outcomes of multiple pregnancies conceived by In vitro fertilization (IVF) and spontaneously and perinatal outcomes of IVF conceived singleton and multiple pregnancies. Additional aim of this thesis is to point out the complexity of this problem and offer possible solutions. Materials and Methods: Design of a study was a combination of retrospective and prospective observational longitudinal cohort study. Analysis included pregnancies which had delivery at the Department of Gynecology and Obstetrics, Clinical Center of Vojvodina in the period from 1.01.2008. to 31.12.2010. The study analyzed and compared the perinatal outcomes in 174 spontaneous conceived multiple pregnancies, 163 multiple pregnancies resulting from IVF procedures, and 155 singleton pregnancies conceived by IVF procedure. Analyzed parameters were newborns birth weight, gestational age at delivery, the value of the Apgar score, occurrence of hypertensive syndrome in pregnancy, gestational diabetes, as well as numerous parameters of perinatal outcome. Taken by trained clinicians and were entered into a specially designed database, the results were statistically analyzed in JMP ver 9.0 software (SAS publisher) using ANOVA analysis to test the statistical significance between the mean values of continuous variables, while the statistical significance of the difference in frequency of categorical variables was assessed by Pearsons &chi;2 test. Results: ART singleton pregnancies had an average mothers age of 33.5 years, the average gestational age at birth of 38.26 gestational weeks (gw), preterm delivery rate of 12.9%, average birth weight 3258 g, Apgar score (AS) in the first minute 8.35, and in the fifth minute 9.2, cesarean section rate 65.81%, Gestational diabetes (GDM) in 7.1% pregnancies, anemia occurred in 41.94% of pregnancies, while preeclampsia was observed in 4.52% of all pregnancies. All relevant parameters of perinatal outcome were significantly (p&lt;0.0001) superior to both ART and non-ART twin pregnancies. ART twin pregnancy showed the average mothers age of 32.9 years, the average gestational age at birth of 35.6 gw, the preterm delivery rate 58.27%, the average body weight newborns 2374 g, AS in the first minute of 7.45, and in the fifth minute of 8.65, the cesarean section rate of 83.7%, GDM in 15.11% of all pregnancies, anemia occurred in 78.42% and preeclampsia in 12.23% of pregnancies, while the non-ART twin pregnancy showed an average mothers age of 28.8 years, the average gestational age at birth of 36.08 gw, the preterm delivery rate of 49.71%, the average body weight of 2433 g, AS in the first minute of 7.75 in the fifth minute 8.75, the caesarian section rate of 58.33%, GDM-a occurred in 7.02%, anemia in 67.84% and preeclampsia in 11.11% of pregnancies. Except for maternal age and the caesarean section rate, which were significantly higher in ART twin pregnancies (p&lt;0.0001), as well as small increase in proportion of amniotic fluid volume disorders (p = 0.033), almost all other parameters of perinatal outcome of were comparable in these groups. Discussion and Conclusion: The study showed that the course and outcome of multiple pregnancies conceived spontaneous and after IVF procedures are equivalent in almost all parameters with similar average body weight and gestational age at birth, and that all these parameters of multiple pregnancies regardless of the conception mode are equally worse compared with singleton pregnancies from IVF procedures. With the exception of multiplicity as a risk factor children from in vitro fertilization procedures are generally healthy. Multiplicity itself and not the mode of conception presented a problem, which is rightly considered the major complication of IVF today. Additional analysis of the experiences of other health system indicates that only a broad and comprehensive implementation of strategy to return only one embryo (SET&ndash;single embryo transfer) can lead to a reduction of the rate of multiple pregnancies after IVF procedures, and the accompanying complications, without compromising IVF success. The experience of other health systems indicate that a successful implementation of SET is only possible with the involvement of the whole society, along with a number of legislative measures in the field of monitoring, control and reimbursement of assisted reproduction procedures. The scope and funding of an IVF procedures (with more free attempts for infertile couples, reimbursed by public health) with mandatory use of SET, and good cryopreservation programs are, based on examples in other countries who had successfully dealt with his problem, is the key in reducing the problem of multiple pregnancies after IVF procedures.</p>