Conséquences du stress en période périnatale sur la réponse émotionnelle chez l'enfant prématuré

Problématique: Les naissances prématurées sont des situations cliniques à risque pour l'enfant. Celui-ci arrive au monde avec une capacité amoindrie à être autonome, que ce soit, par exemple, au niveau respiratoire ou de la régulation thermique. Lors de leur séjour aux soins intensifs, les enfants prématurés subissent des interventions qui peuvent être douloureuses, ils sont souvent manipulés et exposés à des bruits et des lumières intenses; ceci peut induire un stress important pour l'enfant qui est déjà vulnérable du fait de sa prématurité. Lorsque confronté à une situation stressante, l'organisme humain réagit par l'activation de l'axe hypothalamo-hypophysio-surrénalien (HHS). Une des hormones principales sécrétées par l'axe HHS est le cortisol. Plusieurs études suggèrent que le cortisol, lorsque sécrété en quantités importantes, risque d'altérer la matière grise, en induisant une diminution de volume, notamment celui de l'hippocampe (Plotsky & Meaney, 1993; Sapolsky, 2000; McEwen, 1994; 2000; Sandman et al., 1994; Meaney et al., 1991;1996). Ces altérations auraient comme effet un dérèglement de l'axe HHS (Heim & Nemeroff, 1999) avec comme conséquence une variation de la réponse au stress mais aussi de la mémoire et de la régulation émotionnelle (Stam et al., 2000 et Siegel, 1998). Objectifs: Nous allons nous intéresser aux grands prématurés nés à moins de 32 semaines de gestations selon leur exposition à des interventions stressantes et douloureuses ou des complications postnatales. Nous allons étudier l'impact qu'un stress modéré ou sévère sur un prématuré a sur la réponse de l'axe HHS et sur sa régulation émotionnelle lorsque celui-ci est confronté à des situations de stress modéré ultérieurement, plus précisément à six mois d'age corrigé. Méthodologie: Dans le cadre de l'étude «Stress néonatal et réactivité au stress ultérieur: effets préventifs d'une intervention précoce» (SNF 3200 BO-104230) menée au SUPEA en collaboration avec le service de néonatologie du DMCP, le Lab-TAB (Laboratory Temperament Assessment Battery, prelocomotor version 3.1, Goldsmith & Rothbart, 1999) a été utilisé comme outil pour analyser le tempérament d'un enfant par l'analyse de ses réactions émotionnelles à certaines situations. L'enfant, qui était filmé, a été confronté à des mises en scène conçues pour susciter diverses émotions comme la peur, la colère, la frustration, la curiosité ou le plaisir. Un système de codage dérivé de celui du Lab-TAB sera utilisé par deux codeurs différents pour analyser les réactions émotionnelles des enfants. Les taux de cortisol sécrétés ont été mesurés dans la salive de l'enfant. La mesure du cortisol libre dans la salive montre une bonne corrélation avec le cortisol plasmatique ; plusieurs prélèvements de salive ont été faits avant, pendant et après la situation de stress. Les situations filmées n'ont pas encore été codés, ni les taux analysés par l'équipe de la SUPEA. Nous allons observer 40 vidéos, 20 d'enfants nés prématurément et 20 d'enfants nés à terme comme groupe contrôle, et examiner la réponse endocrinienne au stress ainsi que leur régulation émotionnelle au niveau de leur comportement comme l'excitation, les pleurs, l'évitement... Hypothèse : L'intensité de l'exposition au stress postnatal chez l'enfant prématuré induirait des variations de la réponse de l'axe HHS mais aussi de la régulation émotionnelle. Il n'est pas clair selon la littérature actuelle si les variations des réponses de l'axe HHS dues à l'exposition à un stress précoce vont dans les sens de l'hypo- ou l'hyperréactivité, et si cette réponse corrèle avec la régulation émotionnelle au niveau des comportements. C'est ce que l'analyse des données nous permettra de déterminer.

Triplet pregnancies: perinatal outcome evolution

PURPOSE: To evaluate the obstetric and perinatal outcomes evolution of triplet pregnancies.METHODS: A prospective observational study was conducted in triplet pregnancies delivered over 16 years in a tertiary obstetric center with differentiated perinatal support. Evaluation of demographic factors, obstetric complications, gestational age at delivery, mode of delivery, birth weight and immediate newborn outcome were done over a 16 years period. A global characterization of the sample was performed considering the listed parameters. Variables were categorized in three groups according to year of occurrence: 1996-2000, 2001-2006, 2007-2011, and all parameters were compared.RESULTS: Of the 33 triplets included, 72.7% resulted from induced pregnancies. All except one patient received prenatal corticosteroids and five received tocolytics. All women delivered prenatally and no significant differences were seen in the mean gestational age at delivery or birth weight towards time. There were three intrauterine fetal deaths. Neonatal immediate outcomes were not significantly different over the years.CONCLUSION: Despite remarkable progresses in perinatal and neonatal cares, no noticeable impact in triplet gestations' outcomes was seen, sustaining that triplets should be avoided due to their great risk of prematurity and neonatal morbidities, either by limiting the numbers of embryos transferred or by fetal reduction.

Do induced twin pregnancies influence the obstetric and neonatal results of multiple births born before 32 weeks? Comparison to spontaneous gestation

PURPOSE: To compare obstetric outcomes of induced preterm twin births (under 32 weeks gestation) with those spontaneously conceived. METHODS: Prospective study of twin pregnancies (25 induced and 157 spontaneously conceived) developed over a period of 16 years in a tertiary obstetric center. Demographic factors, obstetric complications, gestational age at delivery, mode of delivery, birth weight and immediate newborn outcome were compared. RESULTS: The analysis of obstetrical complications concerning urinary or other infections, hypertensive disorders of pregnancy, gestational diabetes, fetal malformations, intrauterine fetal death, intrauterine growth restriction and intrauterine discordant growth reveal no significant statistical differences between the two groups. First trimester bleeding was higher in the induced group (24 versus 8.3%, p=0.029). The cesarean delivery rate was 52.2% in spontaneous gestations and 64% in induced gestations. Gestational age at delivery, birth weight, Apgar scores at first and fifth minutes, admissions to Neonatal Intensive Care Unit and puerperal complications show no statistically significant differences between the two groups. These results were independent of chorionicity and induction method. CONCLUSION: The mode of conception did not influence obstetric and neonatal outcomes. Although induced pregnancies have higher risk of first trimester bleeding, significant differences were not observed regarding other obstetric and puerperal complications and neonatal results.

Plasma amino acids in pregnancy, placental intervillous space and preterm newborn infants

Plasma amino acid levels have never been studied in the placental intervillous space of preterm gestations. Our objective was to determine the possible relationship between plasma amino acids of maternal venous blood (M), of the placental intervillous space (PIVS) and of the umbilical vein (UV) of preterm newborn infants. Plasma amino acid levels were analyzed by ion-exchange chromatography in M from 14 parturients and in the PIVS and UV of their preterm newborn infants. Mean gestational age was 34 ± 2 weeks, weight = 1827 ± 510 g, and all newborns were considered adequate for gestational age. The mean Apgar score was 8 and 9 at the first and fifth minutes. Plasma amino acid values were significantly lower in M than in PIVS (166%), except for aminobutyric acid. On average, plasma amino acid levels were significantly higher in UV than in M (107%) and were closer to PIVS than to M values, except for cystine and aminobutyric acid (P < 0.05). Comparison of the mean plasma amino acid concentrations in the UV of preterm to those of term newborn infants previously studied by our group showed no significant difference, except for proline (P < 0.05), preterm > term. These data suggest that the mechanisms of active amino acid transport are centralized in the syncytiotrophoblast, with their passage to the fetus being an active bidirectional process with asymmetric efflux. PIVS could be a reserve amino acid space for the protection of the fetal compartment from inadequate maternal amino acid variations.

Morphological and functional characterization of placenta during gestation in bovine clones derived by somatic nuclear transfer

La technique de clonage par transfert nucléaire de cellules somatiques (SCNT) présente une page importante dans les annales scientifiques, mais son application pratique demeure incertaine dû à son faible taux de succès. Les anomalies placentaires et de développement fœtal se traduisent par des pertes importantes de gestation et des mortalités néonatales. Dans un premier temps, la présente étude a caractérisé les changements morphologiques des membranes fœtales durant la gestation clonée en les comparant à des gestations contrôles obtenues à partir de l’insémination artificielle. Les différentes anomalies morphologiques des placentomes telles que l’œdème chorioallantoique, la présence de zones hyperéchoiques et irrégulières dans la membrane amniotique et la présence de cellules inflammatoires dégénérées compromettent le développement fœtal normal de la gestation clonée. L’examen ultrasonographique représente une technique diagnostique importante pour faire le suivi d’une gestation et de caractériser les changements placentaires dans le cadre d’évaluation globale du bien-être fœtal. Le profil hormonal de trois stéroïdes (progestérone (P4), estrone sulfate (E1S), et œstradiol (E2)) et de la protéine B spécifique de gestation (PSPB) dans le sérum des vaches porteuses de clones SCNT a été déterminé et associé aux anomalies de gestations clonées. Une diminution de la P4 sérique au jour 80, une élévation du niveau de la concentration de la PSPB au jour 150, et une augmentation de la concentration d’E2 sérique durant le deuxième et troisième tiers de la gestation clonée coïncident avec les anomalies de gestation déjà reportées. Ces changements du profil hormonal associés aux anomalies phénotypiques du placenta compromettent le déroulement normal de la gestation clonée et gênent le développement et le bien-être fœtal. Sur la base des observations faites sur le placenta de gestation clonée, le mécanisme moléculaire pouvant expliquer la disparition de l’épithélium du placenta (l’interface entre le tissue maternel et le placenta) a été étudié. L’étude a identifié des changements dans l’expression de deux protéines d’adhérence (E-cadhérin et β-catenin) de cellules épithéliales pouvant être associées aux anomalies du placenta chez les gestations clonées. Le tissu de cotylédons provenant de gestations clonées et contrôles a été analysé par Western blot, RT-PCR quantitatif, et par immunohistochimie. Les résultats présentaient une diminution significative (p<0.05) de l’expression des dites protéines dans les cellules trophoblastiques chez les gestations clonées. Le RT-PCR quantitatif démontrait que les gènes CCND1, CLDN1 et MSX1 ciblés par la voie de signalisation de la Wnt/β-catenin étaient significativement sous exprimés. La diminution de l’expression des protéines E-cadherin et β-catenin avec une réduction de l’activation de la protéine β-catenin durant le période d’attachement de l’embryon peut potentiellement expliquer l’absence totale ou partielle de l’attachement des membranes fœtales au tissu maternel et éventuellement, l’insuffisance placentaire caractéristique des gestations clonées chez la vache. La caractérisation morphologique et fonctionnelle du placenta durant les gestations clonées à haut risque est essentielle pour évaluer le statut de la gestation. Les résultats de la présente étude permettront de prédire le développement et le bien-être fœtal de façon critique à travers un protocole standardisé et permettre des interventions médicales pour améliorer le taux de succès des gestations clonées chez les bovins.

Évaluation ultrasonographique du bien-être foetal bovin en fin de gestation

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Placentation and maternal investment in mammals

The mammalian placenta exhibits striking interspecific morphological variation, yet the implications of such diversity for reproductive strategies and fetal development remain obscure. More invasive hemochorial placentas, in which fetal tissues directly contact the maternal blood supply, are believed to facilitate nutrient transfer, resulting in higher fetal growth rates, and to be a state of relative fetal advantage in the evolution of maternal-offspring conflict. The extent of interdigitation between maternal and fetal tissues has received less attention than invasiveness but is also potentially important because it influences the surface area for exchange. We show that although increased placental invasiveness and interdigitation are both associated with shorter gestations, interdigitation is the key variable. Gestation times associated with highly interdigitated labyrinthine placentas are 44% of those associated with less interdigitated villous and trabecular placentas. There is, however, no relationship between placental traits and neonatal body and brain size. Hence, species with more interdigitated placentas produce neonates of similar body and brain size but in less than half the time. We suggest that the effects of placental interdigitation on growth rates and the way that these are traded off against gestation length may be promising avenues for understanding the evolutionary dynamics of parentoffspring conflict. Keywords: placenta, parent-offspring conflict, life history, brain evolution, reproductive strategies, gestation.

Avaliação do papel da ultra-sonografia obstétrica de rotina, no rastreamento pré-natal das anormalidades cardíacas

A ultra-sonografia obstétrica é um método diagnóstico tradicionalmente utilizado na rotina do atendimento pré-natal, tendo sido estudados de forma ampla suas vantagens e limitações. O advento do diagnóstico intra-uterino de cardiopatias congênitas e de arritmias através da ecocardiografia fetal modificou completamente o prognóstico perinatal dessas afecções, por permitir planejar o adequado manejo cardiológico no período neonatal imediato e, em algumas situações, o tratamento e sua resolução in utero. Sendo muito elevada a prevalência de cardiopatias congênitas durante a vida fetal, sua detecção torna-se fundamental. Considerando a inviabilidade operacional de realizar rotineiramente ecocardiografia fetal em todas as gestações, levando-se em conta as condições locais do sistema de saúde, o encaminhamento para exame por especialista passa a ser otimizado com a possibilidade da suspeita de alterações estruturais ou funcionais do coração e do sistema circulatório durante o exame ultra-sonográfico obstétrico de rotina. Não são conhecidos, em nosso meio, dados que avaliem de forma sistemática a acurácia da ultra-sonografia obstétrica no que se refere à suspeita pré-natal de cardiopatias. A partir deste questionamento, este trabalho foi delineado com o objetivo de avaliar o papel da ultra-sonografia obstétrica de rotina na suspeita pré-natal de cardiopatias congênitas ou arritmias graves e os fatores envolvidos na sua efetividade. A amostra foi constituída de 77 neonatos ou lactentes internados no Instituto de Cardiologia do Rio Grande do Sul / Fundação Universitária de Cardiologia (IC/FUC) no período de maio a outubro de 2000, com diagnóstico pós-natal confirmado de cardiopatia estrutural ou arritmia grave, que tenham sido submetidos, durante a vida fetal, a pelo menos uma ultra-sonografia obstétrica após a 18a semana de gestação. Para a coleta de dados, foi utilizado um questionário padronizado, respondido pelos pais ou responsáveis, após consentimento informado. As variáveis categóricas foram comparadas pelo teste do qui-quadrado ou pelo teste de Fisher, com um alfa crítico de 0,05. Um modelo de regressão logística foi utilizado para determinar variáveis independentes eventualmente envolvidas na suspeita pré-natal de cardiopatia. Em 19 pacientes (24,7%), a ultra-sonografia obstétrica foi capaz de levantar suspeita de anormalidades estruturais ou de arritmias. Ao serem consideradas apenas as cardiopatias congênitas, esta prevalência foi de 19,2% (14/73). Em 73,7% destes, as cardiopatias suspeitadas eram acessíveis ao corte de 4-câmaras isolado. Observou-se que 26,3% das crianças com suspeita pré-natal de cardiopatia apresentaram arritmias durante o estudo ecográfico, enquanto apenas 3,4% dos pacientes sem suspeita pré-natal apresentaram alterações do ritmo (P=0,009). Constituiram-se em fatores comparativos significantes entre o grupo com suspeita pré-natal e o sem suspeita a paridade (P=0,029), o parto cesáreo (P=0,006), a internação em unidade de tratamento intensivo (P=0,046) e a escolaridade paterna (P=0,014). Não se mostraram significativos o número de gestações, a história de abortos prévios, o estado civil, o sexo dos pacientes, o tipo de serviço e a localidade em que foram realizados o pré-natal e a ultra-sonografia obstétrica, a indicação da ecografia, o número de ultra-sonografias realizadas, a renda familiar e a escolaridade materna. À análise multivariada, apenas a presença de alteração do ritmo cardíaco durante a ultra-sonografia obstétrica mostrou-se como variável independente associada à suspeita pré-natal de anormalidade cardíaca. Este trabalho demonstra que a ultra-sonografia obstétrica de rotina ainda tem sido subutilizada no rastreamento pré-natal de cardiopatias congênitas, levantando a suspeita de anormalidades estruturais em apenas um quinto dos casos. Considerando a importância prognóstica do diagnóstico intra-uterino de cardiopatias congênitas e arritmias graves, todos os esforços devem ser mobilizados no sentido de aumentar a eficácia da ecografia obstétrica de rotina para a suspeita de anormalidades cardíacas fetais. O treinamento dirigido dos ultra-sonografistas e a conscientização do meio obstétrico e da própria população são instrumentos para esta ação.

Aberrant expression of E-cadherin and beta-catenin proteins in placenta of bovine embryos derived from somatic cell nuclear transfer

Abnormal placental development is common in the bovine somatic cell nuclear transfer (SCNT)-derived fetus. In the present study, we characterised the expression of E-cadherin and beta-catenin, structural proteins of adherens junctions, in SCNT gestations as a model for impaired placentation. Cotyledonary tissues were separated from pregnant uteri of SCNT (n - 6) and control pregnancies (n - 8) obtained by artificial insemination. Samples were analysed by western blot, quantitative RT-PCR (qRT-PCR) and immunohistochemistry. Bovine trophectoderm cell lines derived from SCNT and control embryos were analysed to compare with the in utero condition. Although no differences in E-cadherin or beta-catenin mRNA abundance were observed in fetal tissues between the two groups, proteins encoded by these genes were markedly under-expressed in SCNT trophoblast cells. Immunohistochemistry revealed a different pattern of E-cadherin and total beta-catenin localisation in SCNT placentas compared with controls. No difference was observed in subcellular localisation of dephosphorylated active-beta-catenin protein in SCNT tissues compared with controls. However, qRT-PCR confirmed that the wingless (WNT)/beta-catenin signalling pathway target genes CCND1, CLDN1 and MSX1 were downregulated in SCNT placentas. No differences were detected between two groups of bovine trophectoderm cell lines. Our results suggest that impaired expression of E-cadherin and beta-catenin proteins, along with defective beta-catenin signalling during embryo attachment, specifically during placentation, is a molecular mechanism explaining insufficient placentation in the bovine SCNT-derived fetus.

Somatic cell nuclear transfer is associated with altered expression of angiogenic factor systems in bovine placentomes at term

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of age at first calving, gestation length and dry period on milk yield in a Gyr herd

Relationships of several reproductve traits and milk yield were studied in 716 Gyr cows at Sant'Ana da Serra farm, Mococa, State of São Paulo, a humid tropical climate region. Mean age at first calving was 49.8 +/- 0.4 months, with a coeficient of cariation (C. V.) of 20.8%. Only year of parturition significantly affected age at first calving (P < 0.01). Heritability, estimated from paternal half-sib correlations, was 0.91 +/- 0.20, a value considered unrealistically high. Overall mean gestation length was 287.7 +/- 0.5 days (C. V. = 3.3%) for 419 observations, with month and year of parturition having significant effects. Heritability estimate was 0.30 +/- 0.14. Overall mean dry period for 1.276 observations was 238 +/- 3 days (C. V. = 48%); repeatability estimate was 0.19 +/- 0.06. Estimated annual genetic trend for dry period was -0.6 days, phenotypic trend was 11.5 days, and environmental trend, 12.1 days. Month and year of parturition and cow age had no significant effect on dry period. Highest milk yield was obtained at fourth lactation. The gross correlation between milk yield and gestation period was 0.11, and between previous dry period and subsequent milk yield, 0.16. Normal gestations of 285 to 290 days were associated with higher milk yields. Milk yield increased as dry period advanced from 30 to 390 days, and declined as dry period continued beyond 390 days. Delays in first mating and a more extensive dry period decreased reproductive efficiency in the herd studied.

Uterine evaluation and gestation diagnosis in owl monkey (Aotus azarai infulatus) using the B mode ultrasound

Background Gynecological and obstetrical ultrasonography has become an indispensable tool in the routine management, health evaluation and research on captive non-human primates.Methods Ultrasound was used to evaluate the uterus and estimate the gestation of owl monkeys. Twelve couples were selected, where five were primiparous and seven multiparous females from the National Primate Center reproductive colony, Ananindeua-PA, Brazil. The procedures were carried out using the GE (R) Logiq 100 MP, equipped with a 7.5 MHz linear probe.Results the females showed a simple uterus, of elongated shape, regular outline and homogeneous echogenic texture. In the uterine measurements craniocaudal diameter, dorsoventral diameter and uterine volume (UV), significant differences were identified (P < 0.05) between ultrasound examinations of primiparous and multiparous females. The UV showed a positive correlation with the number of births. The gestational sac and the embryonic echo were visible between 28 and 38 days after mating. Between 48 and 68 days after mating, embryonic death was identified in all the gestations.Conclusions the chemical (use of tranquilizers) and husbandry factors (capture stress) may be related to the prenatal death. The establishing methods of conditioning the female to the ultrasonographic exam may offer a solution to this problem.

Caracterização da fusão caruncular em gestações naturais e de conceptos bovinos clonados

Barreto R.S.N., Miglino M.A., Meirelles F.V., Visintin J.A., Silva S.M., Burioli K.C., Fonseca R., Bertan C., Assis Neto A.C. & Pereira F.T.V. 2009. [Characterization of the caruncular fusion in gestations of natural and cloned bovine conceptuses.] Caracterizacao da fusao caruncular ern gestacoes naturais e de conceptos bovinos clonados. Pesquisa Veterinaria Brasileira 29(10):779-787. Laboratorio de Morfofisiologia da Placenta e Embriao, Faculdade de Zootecnia, Universidade Estadual Paulista, SP294 Km 651, Dracena, SP 17900-000, Brazil. E-mail: fverechia@dracena.unesp.brThe objective of the study was to compare the characteristics of the caruncular fusion in gestations of non-cloned and cloned conceptuses. The non-cloned conceptuses were divided according to the gestation period: Group 1 (2 to 3 months; n=9),II (4 to 6; n=9); III (7 to 8; n=10) and IV (9 n=7). The cloned conceptuses formed the Group V: 9 months; n=4. The caruncles were observed macroscopically (number and dimensions: length, width and height), microscopically and submitted to statistical analysis (5% of significance). We observed three types of macroscopic caruncular fusions: oval (morphologically normal); two united adjacent caruncles and the lobulated type, characterized by regions with several united caruncles presenting a false fusion or deformation of the caruncular parenchyma. The length of the caruncles was 1.55 +/- 0.57; 2.45 +/- 0.55; 4.66 +/- 2.0 and 5.72 +/- 1.90cm for the groups 1, 11, 111, IV respectively. As for the height, the caruncles presented a lineal growth during the gestation: 0.40 +/- 0.15; 0.57 +/- 0.21; 1.0 +/- 0.48 and 1.80 +/- 0.91cm, for the respective groups 1, 11, 111 and IV. The width of the caruncles was similar between the groups I and 11 (0.97 +/- 0.30 e 1.42 +/- 0.71 cm) and the groups III and IV (2.68 +/- 1.22 and 3.52 +/- 1.16cm). When the group V was compared to the IV, the caruncles of the group V presented a larger length (5.72 +/- 1.90 vs. 7.88 +/-.13cm) and width (3.52 +/- 1.16 vs. 4.93 +/- 1.46cm), however they were similar in height (1.80 +/- 0.91 and 2.25 +/- 0.67cm). We verified that in gestations of cloned conceptuses the caruncles presented a larger development than in gestations of non-cloned conceptuses. The fusioned caruncles presented measurements statistically similar to the isolated ones in all the parameters and groups. Under light microscopy, we observed the formation of a stromal axis from the basis of the caruncle to the apex of the fusional fissure, with the histological constitution similar to the endometrial stroma. Three microscopic shapes were also unpublished defined: true fusion with a single axis evident below the fusional fissure; pseudofusion with a double axis in H shape and false fusion with absence of the axis. The first two formats were associated to the oval and lobulated caruncles and the last one to the false fusion with deformation of the caruncle parenchyma. The fusional axis increased in size along the gestation among the groups I, II, III and IV. The group V presented a larger length and width of the axis when compared to the group IV. Thus, in gestations of cloned conceptuses a destruction of the lateral epithelium of the caruncles is associated to an incompetence in the maternal-fetal interdigitation, that compromises the cotyledonary fusion. We suggest that, in gestations derived of cloned conceptuses, the increase of the size of the caruncular fusions is possibly associated to a compensatory mechanism for the metabolic exchanges between mother and fetus, in reason of the smallest number of isolated caruncles.

Incidência de malformações congênitas em crianças concebidas através de injeção intracitoplasmática de espermatozóides

Purpose: to evaluate the incidence and types of major congenital malformations (MCM) in liveborn children conceived by intracytoplasmic sperm injection (ICSI). Methods: a total of 680 liveborn children resulted from 511 couples submitted to ICSI from January, 1999 to December, 2002. Data collection of the children was performed through standardized questionnaire and clinical examination. Of the 511 couples, 366 had been contacted for a sampling of 371 gestations. Of the 680 liveborn, 520 had been evaluated, 250 of them (48.1%) through questionnaire and 270 (51.9%) through questionnaire and physical examination. Two hundred and fifty children were from singleton pregnancies and 270 from multiple pregnancies. Malformations were classified according to the 10th revision of the International Statistical Classification of Diseases and Related Health. Only MCM were analyzed in this study. The incidence of MCM was compared with that of the general population obtained by the Latin American Collaborative Study of Congenital Malformations. The statistical analysis was performed by the χ 2 test (level of significance p<0.05). Results: of the 520 children, 15 presented MCM, resulting in an incidence of 2.9%. There was no difference in relation to the control group (p>0.05), which showed 2.6% incidence of MCM. The most frequent malformations were of cardiac origin (four isolated and two associated), corresponding to 40% of the total. The other types of MCM were: renal (three), neural tube (two), skull (one), cleft lip (one), genital (one), Down syndrome (associated with cardiac malformations) (two), and musculoskeletal (one). Six MCM occurred in children from singleton pregnancies and nine in children from multiple pregnancies. Conclusion: the liveborn children conceived by ICSI presented incidence of major congenital malformations (2.9%) near to the expected for the general population (2.6%). However, to establish the risks of MCM with precision it is necessary to continue the evaluation of the children conceived by ICSI.

Reproductive biology of the guitarfish Rhinobatos percellens (Chondrichthyes, Rhinobatidae) from the São Paulo Coast, Brazil, western South Atlantic Ocean

The reproductive biology of the guitarfish Rhinobatos percellens was studied from 751 specimens caught by bottom pair trawlers off the coast of São Paulo, Brazil, between c. 24° 00′ S; 45° 15′ W and c. 25° 10′ S; 47° 52′ W, from September 2007 to August 2009. The total length (LT) and total mass (MT) relationship for males and females combined was MT = 1·29E-06 LT 3·15 (r = 0·99, n = 751). The mean LT of sexually mature specimens was 548 mm for males and 583 mm for females. Clasper growth was allometric and showed three distinct phases. Most claspers were calcified in specimens of c. 550 mm LT. The mean diameter of the largest oocyte was 29·8 mm, the mean ovarian fecundity was seven oocytes and ovulation occurred between August and November. Uterine fecundity ranged from two to 13 embryos (mean of five embryos). Larger females had higher litter sizes and larger embryos; the size-at-birth was c. 200 mm LT. The hepato-somatic index oscillated seasonally for males and females; the gonado-somatic index had little variation in males, but varied seasonally in females. The presence of many non-pregnant adult females and of encapsulated eggs during two consecutive seasons suggests a resting period between gestations and the possibility of diapause. © 2012 The Authors. Journal of Fish Biology © 2012 The Fisheries Society of the British Isles.