Regular physical activity during pregnancy provides both maternal and infant health benefits. The complexity of measuring physical activity during pregnancy hampers the determination of the optimal dose of habitual physical activity for pregnant women and has led to broad physical activity guidelines for pregnant women. Subjectively-determined physical activity levels by means of questionnaires may have contributed to these broad guidelines. However the ActiHeart®, a dual heart rate monitor and accelerometer, is an accurate and reliable measurement tool to determine physical activity levels during pregnancy. Maternal physical activity tends to decrease during pregnancy and may lead to various health risks, including excessive weight gain, risk for gestational diabetes, lower back pain and adverse foetal outcomes. Determining the influence of physical activity on foetal growth is confounded by various variables, therefore objectively-measured habitual physical activity is essential. This study aims to objectively determine habitual physical activity patterns of pregnant women and the relationship between habitual physical activity and foetal growth parameters. In a longitudinal, observational, cohort study design, 60 pregnant women were measured at four stages in their pregnancy: the first trimester (9 – 12 weeks), second trimester (20 – 22 weeks), third trimester (28 – 32 weeks) and three months postpartum. Demographic information was collected by means of a questionnaire specifically compiled for this study, followed by anthropometric measurements (height and weight). Assessment of the participants resting blood pressure, heart rate (Microlife® Semi-Automatic blood pressure and heart rate monitor) and metabolic rate (FitmateTM, Cosmed) was obtained. Thereafter, a step-test was performed for individualised calibration of the ActiHeart® device for assessment of habitual physical activity patterns over a 7-day period. Foetal growth parameters that included birth weight (kg), birth length (cm), abdominal circumference (cm) and head circumference (cm), were collected from medical records and from the mother post-partum. Habitual physical activity, presented as average Activity Energy Expenditure (AEE), physical Activity Level (PAL), activity counts and minutes spent in activity, declined from the first to the third trimester of pregnancy. The AEE during the first trimester averaged 803 ± 34 kCal/day and declined statistically significant to 592 ± 383 kCal/day in the third trimester. Minutes spent per week doing moderate activity declined from 103 ± 83 min/week in the first trimester to 55 ± 66 min/week in the third trimester. Average pregnancy AEE indicated a non-significant negative relationship with all foetal growth measurements - birth weight (r = - 0.39, p = 0.45), birth length (r = - 0.16, p = 0.77), Ponderal Index (r = - 0.34, p = 0.51) - and a non-significant positive relationship with head circumference at birth (r = 0.14, p = 0.79). In conclusion, the objectively-determined, habitual physical activity levels of the participants did not meet the stated guidelines for pregnant women. During the progression of pregnancy, the activity levels declined significantly at the third trimester. The habitual activity levels indicate no effect on the foetal growth parameters. / MSc (Biokinetics), North-West University, Potchefstroom Campus, 2014

Regular physical activity during pregnancy provides both maternal and infant health benefits. The complexity of measuring physical activity during pregnancy hampers the determination of the optimal dose of habitual physical activity for pregnant women and has led to broad physical activity guidelines for pregnant women. Subjectively-determined physical activity levels by means of questionnaires may have contributed to these broad guidelines. However the ActiHeart®, a dual heart rate monitor and accelerometer, is an accurate and reliable measurement tool to determine physical activity levels during pregnancy. Maternal physical activity tends to decrease during pregnancy and may lead to various health risks, including excessive weight gain, risk for gestational diabetes, lower back pain and adverse foetal outcomes. Determining the influence of physical activity on foetal growth is confounded by various variables, therefore objectively-measured habitual physical activity is essential. This study aims to objectively determine habitual physical activity patterns of pregnant women and the relationship between habitual physical activity and foetal growth parameters. In a longitudinal, observational, cohort study design, 60 pregnant women were measured at four stages in their pregnancy: the first trimester (9 – 12 weeks), second trimester (20 – 22 weeks), third trimester (28 – 32 weeks) and three months postpartum. Demographic information was collected by means of a questionnaire specifically compiled for this study, followed by anthropometric measurements (height and weight). Assessment of the participants resting blood pressure, heart rate (Microlife® Semi-Automatic blood pressure and heart rate monitor) and metabolic rate (FitmateTM, Cosmed) was obtained. Thereafter, a step-test was performed for individualised calibration of the ActiHeart® device for assessment of habitual physical activity patterns over a 7-day period. Foetal growth parameters that included birth weight (kg), birth length (cm), abdominal circumference (cm) and head circumference (cm), were collected from medical records and from the mother post-partum. Habitual physical activity, presented as average Activity Energy Expenditure (AEE), physical Activity Level (PAL), activity counts and minutes spent in activity, declined from the first to the third trimester of pregnancy. The AEE during the first trimester averaged 803 ± 34 kCal/day and declined statistically significant to 592 ± 383 kCal/day in the third trimester. Minutes spent per week doing moderate activity declined from 103 ± 83 min/week in the first trimester to 55 ± 66 min/week in the third trimester. Average pregnancy AEE indicated a non-significant negative relationship with all foetal growth measurements - birth weight (r = - 0.39, p = 0.45), birth length (r = - 0.16, p = 0.77), Ponderal Index (r = - 0.34, p = 0.51) - and a non-significant positive relationship with head circumference at birth (r = 0.14, p = 0.79). In conclusion, the objectively-determined, habitual physical activity levels of the participants did not meet the stated guidelines for pregnant women. During the progression of pregnancy, the activity levels declined significantly at the third trimester. The habitual activity levels indicate no effect on the foetal growth parameters. / MSc (Biokinetics), North-West University, Potchefstroom Campus, 2014

The imprinted expression of the IGF2 and H19 genes is controlled by the imprinting control region 1 (ICR1) located at chromosome 11p15.5. This methylation-sensitive chromatin insulator works by binding the zinc-finger protein CTCF in a parent-specific manner. CTCF binds the unmethylated maternal allele and is required for preventing de novo methylation at ICR1. DNA methylation defects involving the ICR1 IGF2/H19 domain result in two growth disorders with opposite phenotypes: an overgrowth disorder, the Beckwith-Wiedemann syndrome (ICR1 gain of methylation in 10% of BWS cases) and a growth retardation disorder, the Silver-Russell syndrome (ICR1 loss of methylation in 60% of SRS cases). Little information is available regarding the mechanism of ICR1 DNA methylation defects. Several deletions removing part of ICR1 (1.4 to 2.2 kb) have been described in a few familial BWS cases with dominant maternal transmission. In order to evaluate precisely the incidence of ICR1 mutations, we investigated, by long range PCR and sequencing, 21 BWS patients (including two brothers) with ICR1 gain of methylation and 16 SRS patients with ICR1 loss of methylation. No mutation of the seven CTCF binding sites was detected in the familial BWS cases. Two additional cases of constitutional genetic lesions were identified in BWS patients with apparently-sporadic forms. One patient was identified with a 8 bp deletion within the B3 repeat, 116 bases 3' of the CTCF binding site 4. Another patient was identified with a 1.8 kb deletion which eliminates CTCF binding sites 2 and 3. A single-nucleotide variation was identified in a SRS patient. Our data showed that ICR1 deletions, including new small deletions, account for apparently sporadic forms of BWS with ICR1 gain of methylation. ICR1 deletions are associated with a high incidence of Wilms' tumour, making their molecular diagnosis particularly important for genetic counseling and tumor surveillance.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第22640号 / 医博第4623号 / 新制||医||1044(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 近藤 玄, 教授 斎藤 通紀, 教授 篠原 隆司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Contexte. La chloration est le traitement le plus utilisé pour désinfecter l’eau distribuée à la population. Lors du traitement, des sous-produits de chloration (SPCs) se forment par réaction de la matière organique naturelle présente dans l’eau avec le chlore. Les produits les plus présents sont les trihalométhanes (THMs), les acides haloacétiques (HAAs). Des études toxicologiques (animales et in vitro) ont montré la génotoxicité et/ou carcinogénicité et la reprotoxicité de certains SPCs. Nous avons mené deux études évaluant les effets sur la santé humaine de l’exposition aux SPCs, dans le domaine du cancer et de la reproduction. Association entre exposition aux THMs et risque de cancer de la vessie : une analyse poolée de 3 études cas-témoins européennes. Cette analyse a inclus 2381 cas et 3086 témoins issus de 3 études cas-témoins (France, Espagne, Finlande). L’exposition environnementale aux SPCs a été mesurée par la concentration en THMs estimée rétrospectivement dans les réseaux de distribution d’eau au cours des 40 années de la fenêtre d’exposition. Les usages de l’eau connus sont l’ingestion, les douches et les bains, la fréquentation de piscine (étude espagnole seulement). Une relation croissante a été observée entre le niveau environnemental de THMs, la durée d’exposition à une eau de surface chlorée et le risque de cancer de la vessie, chez les hommes uniquement. Aucune association n’a été observée avec l’exposition via l’ingestion d’eau du robinet. L’exposition via les douches, les bains et la piscine est apparue liée au risque de cancer de la vessie. Trois études cas-témoins nord-américaines ont été intégrées dans une méta-régression. Aucune spécificité européenne de la relation dose-réponse n’a été mise en évidence. Une relation dose-réponse globale incluant 4351 cas et 7055 témoins a été estimée. Association entre exposition aux SPCs et risque de prématurité et de retard de croissance intra-utérin (RCIU). Cette étude est issue de la cohorte bretonne Pélagie (3400 femmes enceintes recrutées en début de grossesse entre 2002 et 2006). L’exposition pendant la grossesse a été mesurée à l’aide de 2 indicateurs : la concentration en THMs de l’eau distribuée dans les réseaux et le dosage d’un biomarqueur urinaire (acide trichloroacétique) pour un sous-échantillon de femmes (étude cas-témoins nichée). Les niveaux dans les réseaux proviennent de la base de données réglementaire SISE-Eaux. Les usages de l’eau du robinet par les femmes pendant la grossesse ont été collectés par questionnaire (quantité d’eau du robinet bue, fréquentation de la piscine, fréquence et durée des douches et des bains). Notre étude suggère une association entre l’exposition prénatale aux SPCs et le risque de RCIU. Aucune association n’est observée avec le risque de prématurité. / Background. Chlorine is the most widely used disinfectant for drinking water treatment. During treatment process, the natural organic matter present in water reacts with chlorine to form disinfection by-products (DBPs), such as trihalomethanes (THMs) and haloacetic acids (HAAs), the most occurring ones. Several identified DBPs were recognized carcinogens or fetotoxic in animals. We conducted two epidemiological studies to assess the health effects of human exposure to DBPs regarding cancer and reproduction. Association between THM exposure and the risk of bladder cancer: a pooled analysis of 3 European case-control studies. The study included 2381 cases and 3086 controls from 3 case-control studies (France, Finland, Spain). Environmental exposure to SPCs was measured by THM concentrations in water distribution systems, retrospectively estimated through the 40-year exposure window. Information about water uses (tap water ingestion, showers and baths, swimming-pool attendance) was available. An increased risk of bladder cancer was observed, in men only, with increasing THM concentrations in water systems and duration of exposure to chlorinated surface water. No association was observed with exposure through tap water ingestion. Exposure through showers, baths and swimming in pools (available in Spanish study only) was associated with an increased risk of bladder cancer. Three North-American case-control studies were joined to the three European ones into a meta-regression. No specific dose-response relationship was identified for European studies. A global dose-response relationship was consequently estimated, including 4351 cases and 7055 controls. Association between DBP exposure and the risk of preterm birth and Intra-Uterine Growth Retardation (IUGR). This study was conducted within the PELAGIE cohort (3400 pregnant women included in early pregnancy, in Brittany, France, between 2002 and 2006). Exposure during pregnancy was measured by 2 indicators: THM concentrations in water distribution systems and a biomarker of HAA ingestion, the level of trichloroacetic acid in maternal urine at inclusion (in a nested case-control study). The regulatory database SISE-Eaux was used to estimate THMs concentrations in the water distribution systems. Water uses during pregnancy (ingested tap water, swimming-pool attendance, showers and bath frequencies and duration) were collected with questionnaires. Our results suggest an association between prenatal exposure to DBPs (THMs and HAAs) and the risk of IUGR. No association was found with the risk of preterm birth.

Tesis por compendio / [ES] Para evaluar los efectos del proceso de selección en una línea paterna de conejos, se compararon los rasgos de crecimiento y rendimiento reproductivo de la descendencia actual (generación R36) en similar ambiente con una población control rederivada de embriones de una generación anterior (R18). Para reducir/evitar el efecto del proceso de criopreservación sobre los rasgos fenotípicos se rederivaron embriones de la generación actual (R36) y obtener una 3ra población (R37V). Capítulo 1, se compara la R37V y la descendencia de la R36 nacida por inseminación artificial (R37). Hubo diferencias en rasgos de crecimiento postnatal en las 3 generaciones evaluadas, pero no en el crecimiento fetal, componentes del tamaño de la camada y rasgos reproductivos. Así, los procesos de criopreservación y transferencia de embriones causan cambios en rasgos de crecimiento de poblaciones reconstituidas que influyen en las siguientes generaciones, sin producir cambios en los reproductivos. En los siguientes capítulos, para reducir/evitar los efectos del proceso de rederivación, la deriva genética y factores ambientales sobre los rasgos fenotípicos, se usó sólo poblaciones rederivadas para comparar los rendimientos reproductivos y de crecimiento. Capítulo 2, al ser los machos usados para producir dosis seminales en centros de inseminación y granjas, se evaluó si el programa de selección modificaba los rasgos seminales, el proteoma del plasma seminal y de espermatozoides, y la fertilidad del semen al usarla en inseminación artificial. Se analizó el proteoma del plasma seminal y espermatozoides de machos maduros de c/grupo y se prepararon dosis seminales para inseminación. Sólo el porcentaje de espermatozoides anormales mostró diferencias, presentando los R21 menos espermatozoides anormales que los R39. El análisis discriminante (DA-PLS) mostró efecto de la generación para el proteoma plasmático y espermático. En plasma seminal, se reportaron 64 proteínas diferencialmente expresadas y 56 sobreexpresadas en R39 (87,5%). Del proteoma de espermatozoides 132 diferencialmente abundantes y 89 sobreexpresadas en R39 (67,4%). A pesar de las diferencias en importantes proteínas relacionadas con la capacitación, la motilidad, la inmunoprotección de espermatozoides y la fecundación, no hubo diferencias en fertilidad y prolificidad con las dosis seminales para inseminación. Capítulo 3, se analizó el efecto de la selección por ganancia media diaria de peso (GMD) post-destete después de 37 generaciones. Tras 2 generaciones post rederivación (R21 vs. R39), todos los caracteres evaluados mostraron progreso resultado de la selección, y no afecta a los parámetros estimados de la curva de crecimiento de Gompertz. Los resultados demuestran que el programa de selección mejoró la GMD sin variar el peso corporal adulto, pero tras 37 generaciones de selección, este carácter parece agotado. Capítulo 4, comparamos los parámetros reproductivos de conejas entre las poblaciones rederivadas y control. Los rasgos de desarrollo prenatal y los componentes del tamaño de camada se midieron en la 2da generación post rederivación (R20 y R38). Así la selección por GMD no tiene efectos adversos sobre los componentes del tamaño de la camada, y el área del saco fetal al día 12 de gestación, área de la placenta fetal y la longitud cráneo-rabadilla del feto al 19 de gestación fueron mayores en R38. Resultados muestran que la selección por GMD no afecta negativamente el rendimiento reproductivo. Conclusión, el estudio demuestra que los efectos de la criopreservación sobre los rasgos de crecimiento persisten dos generaciones post rederivación. Además, la línea muestra signos de agotamiento del progreso genético quizás por el bajo rendimiento reproductivo y elevada mortalidad postnatal. La selección por GMD influyó en cambios del crecimiento fetal y del proteoma del eyaculado, sin afectar al rendimiento reproductivo de hembras ni la fertilidad y prolificidad de las dosis seminales de machos. / [CA] Per avaluar els efectes del procés de selección en una línia paterna de conills, es van comparar els trets de creixement i el rendiment reproductiu de la descendència de la generació actual (R36) sota el mateix entorn amb una població control derivada dels embrions emmagatzemats d'una generació anterior (R18). Per reduir/evitar l'efecte del procés de criopreservació embrions de la generació actual (R36) es van criopreservar i transferir (rederivar) per obtenir una 3a població (R37V). Capítol 1, es compara la generació R37V i la descendència de la 36a generació nascuda per inseminació artificial (R37). Hi ha diferències en els trets de creixement postnatal a les tres generacions avaluades. Tot i que el creixement fetal, els components de la mida de la ventrada i els trets reproductius no van mostrar diferències. La rederivació provoquen canvis en els trets de creixement de les poblacions reconstituïdes que influeixen en les generacions següents, sense canvis en els trets reproductius. En els capítols següents, per reduir/evitar els efectes del procés de rederivació, la deriva genètica i els factors ambientals sobre els trets fenotípics, es van utilitzar les poblacions rederivades per comparar els rendiments reproductius i de creixement. Capítol 2, ja que els mascles s'utilitzaven per produir dosis de semen en centres d'inseminació i granges, es va estudiar si un programa de selecció per guany mitja de pes diari pot canviar els trets seminals, el proteoma del plasma i dels espermatozoides i la fertilitat del semen en la inseminació artificial. Només el percentatge d'espermatozoides anormals mostra diferències significatives, R21 presentant menys espermatozoides anormals que R39. L'anàlisi discriminant (DA-PLS) mostra efecte de la generació al proteoma del plasma i dels espermatozoides. En plasma seminal, 64 proteïnes es van expressar de manera diferent, 56 sobre-expressades en R39 (87,5%). El proteoma de l'esperma 132 proteïnes diferencialment abundants, 89 sobre-expressades en R39 (67,4%). Tot i observar diferències en proteïnes importants relacionades amb la capacitat, la motilitat o la immunoprotecció dels espermatozoides i la fecundació, La fertilitat i la prolificitat es van detectar quan es van utilitzar dosis seminals comercials per a la inseminació. Capítol 3, vam avaluar l'efecte d'una selecció a llarg termini per a l'augment de pes mitjà diari (ADG). Després de dues generacions d'ambdues poblacions rederivades (R21 vs. R39), tots els trets avaluats mostra algun progrés. Aquesta resposta no sembla afectar els paràmetres estimats de la corba de creixement de Gompertz. Resultats demostra que el programa de selecció havia millorat l'ADG sense variacions en pes corporal adult, però després de 37 generacions de selecció, aquest tret sembla esgotat. Capítol 4, compara els trets reproductius entre poblacions femenins entre ambdues poblacions (rederivades i control). El desenvolupament fetal i els components de la mida de la ventrada es mesura els trets a la segona generació després de la rederivació (R20 i R38). Resultats suggereix que la selecció per ADG no té cap efecte advers sobre els components de la mida de la ventrada i la zona del sac fetal el dia 12 de gestació i la zona de la placenta fetal i la longitud de la corona-gropa del fetus el dia 19 de gestació eren més grans a la R38. La selecció per ADG no afecta negativament els components de la mida de la ventrada, el creixement fetal i el rendiment reproductiu. Nostre estudi proporciona més proves dels efectes de la criopreservació sobre els trets de creixement que persisteixen dues generacions després de la rederivació. La línia va mostrar signes d'esgotament del progrés genètic per baix rendiment reproductiu i l'alta mortalitat postnatal. La selecció per ADG va influir en els canvis en el creixement fetal i en el proteoma ejaculat, però no va afectar el rendiment reproductiu de les femelles ni la fertilitat i la prolificitat de les dosis seminals dels mascles. / [EN] To evaluate the effects of the selection process in a paternal line of rabbits, growth traits and reproductive performance from the offspring of the current generation (R36) were compared under the same environment with a control population rederived from embryos stored of a previous generation (R18). To reduce or avoid the effect of the cryopreservation process on phenotypic traits embryos of current generation (R36) were cryopreserved and transferred to obtain a third population (R37V). In chapter 1, R37V generation and offspring of 36th generation born by artificial insemination (generation R37) were compared. Differences in postnatal growth traits were observed in the three generations assessed. Although foetal growth, litter size components and reproductive traits did not show significant differences. In conclusion, cryopreservation and embryo transfer processes cause changes in growth traits of reconstituted populations that influence the following generations, without changes in reproductive traits. In the following chapters, to reduce or avoid the effects of the rederivation process, genetic drift and environmental factors on phenotypic traits, only the rederived populations were used to compare reproductive and growth performances. In chapter 2, considering that males were used to produce semen doses at insemination centres and farms, we studied whether a programme of selection by daily gain changed the seminal traits, plasma and sperm proteome and the fertility of semen when used in artificial insemination. Seminal plasma and sperm proteome from mature males of each group were analysed and semen doses were used to inseminate females. Only the percentage of abnormal sperm showed significant differences, R21 presented fewer abnormal sperm than R39. The discriminant analysis (DA-PLS) showed an effect of the generation for plasma and sperm proteome. In seminal plasma, 64 proteins were differentially expressed, 56 were overexpressed in R39 (87.5%). Sperm proteome reported 132 differentially abundant proteins, 89 were overexpressed in R39 (67.4%). Despite differences in important proteins related to capacitation, sperm motility or immunoprotection and to the fertilization process, no differences in fertility and prolificacy were detected when commercial seminal doses were used for insemination. In chapter 3, we evaluated the effect of a long-term selection for post-weaning average daily weight gain (ADG) over 37 generations. After two generations of both rederived populations (R21 vs. R39 generations), all evaluated traits showed some progress as a result of the selection. This response does not seem to affect the estimated Gompertz growth curve parameters. Results demonstrated that the selection programme had improved ADG without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. In chapter 4, we compared female reproductive traits between both rabbit populations (rederived and control). Foetal growth and litter size traits were measured in the second generation after rederivation (R20 and R38 generations). Our study suggests that selection for growth rate has no adverse effect on litter size components and the foetal sac area at day 12 of gestation, and foetal placenta area and crown-rump length of the foetus at day 19 of gestation were higher in the R38 generation. These results show that selection for growth rate does not adversely affect on reproductive performance. In conclusion, our study provides further evidence of the effects of cryopreservation on growth traits persisting two generations after rederivation. Moreover, the paternal line showed signs of genetic progress exhaustion due to low reproductive performance and high postnatal mortality. Selection by daily weight gain influenced changes in foetal growth and ejaculate proteome, but did not affect the reproductive performance of females or the fertility of seminal doses of males. / This research was supported by AGL2017-85162-C2-1-R research project funded by Ministerio de Economía, Industria y Competitividad (MICINN, Spain). Funding for open access charge: CRUE- Universitat Politècnica de València. / Juárez Moreno, JD. (2022). Embryo cryopreservation and transfer to rederive a paternal rabbit line after 18 generations. Evaluation of growth and reproductive traits [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/185071 / TESIS / Compendio