Pregnancy diagnosis by transrectal ultrasonography (using a 5 MHZ linear probe) presented the maximum sensitivity and negative predictive values at day 26 and day 29 after estrus in heifers and cows, respectively. Palpation per rectum using the fetal membrane slip for pregnancy diagnosis did not increase embryo/fetus mortality when compared with a positive control group of non-palpated females. The use of a controlled randomized block design was a useful approach to study this problem. Blocking for category and number of embryos allowed us to remove these confounding factors. Factors that affected pregnancy loss during the first four months of pregnancy were: period of pregnancy, age of the animal, number of previous lactations and number of embryos. Pregnancy loss was higher during the embryonic than fetal periods. Spontaneous embryo/fetal mortality increased with the age of the animal and lactation number. The risk of spontaneous embryo/fetus mortality was higher in twin than in single pregnancies. Two types of embryo/fetus mortality were noted: Type I and Type II. Type I was characterized by presence of positive fetal membrane slip by palpation per rectum, signs of degeneration by transrectal ultrasonography and persistence of a functional corpus luteum. The uterus took approximately 3 weeks to be noted clean by transrectal ultrasonography and the animals showed estrus one month after the conceptus was diagnosed dead. Type II was characterized by absence of positive signs of pregnancy by palpation per rectum, absence of signs of degeneration by transrectal ultrasonography and absence of a functional corpus luteum. Pregnancy loss in nuclear transfer derived embryos was higher compared to in vivo derived embryos produced by artificial insemination. Pregnancy loss occurred mainly during the transition from the embryonic to the fetal period. Embryo/fetus mortality detected was Type I. Progesterone produced by the corpus luteum was noted at pregnancy levels for approximately two weeks after embryo/fetus death. Protein B, a hormonal placental marker, was maintained at pregnancy levels for approximately 3 weeks after embryo/fetus death. No differences in the levels of the two hormones were noted when comparing females with dead or live conceptuses.

Homozygous α0-thalassemia is a serious autosomal recessive disorder with poor fetal outcome and severe maternal complications. Conventionally, prenatal diagnosis is performed by an invasive test. A non-invasive approach using serial ultrasonography can effectively reduce the need for invasive tests in unaffected pregnancies. For two-dimensional ultrasound prediction, a total of 777 at-risk fetuses were studied from 12 to 20 weeks between 1995 and 2006. At 12–15 weeks’ gestation, the highest sensitivity (98.3%) was achieved by the combination of fetal cardiothoracic ratio (CTR) and/or middle cerebral artery peak systolic velocity (MCA-PSV) at a false-positive rate of 15.8%. At 16–20 weeks’ gestation, the sensitivity of CTR was 100.0%, but the false-positive rate was 5.2%. In contrast, the false-positive rate of MCA-PSV alone was 1.4% and that of the combination of CTR and MCA-PSV was 0%, although their sensitivities were less than 65%. In a cross-sectional retrospective study of 546 samples at-risk and control (268 fetal and 278 neonatal cord blood), the degree of anemia was only mild in 27.5% of the affected fetuses (see chapter 3 for definition of mild anemia). Because MCA-PSV is not very predictive of mild anemia, this may be one of the reasons why MCA-PSV is not very sensitive in predicting an affected pregnancy. A total of 832 at-risk pregnancies were studied using same noninvasive approach at Maternal and Neonatal Hospital of Guangzhou (MNH) and Tsan Yuk Hospital (TYH). The overall sensitivity and specificity of the noninvasive approach was 100% and 95.6% respectively. At MNH, the need for an invasive test was reduced by 78.6%, and all the affected pregnancies were diagnosed before 24 weeks’ gestation. After adequate training and monitoring the quality of the subsequent ultrasound examinations, the results achieved at MNH were comparable to TYH, with at-risk pregnancies including the affected ones being seen at a more advanced gestation at MNH. In a retrospective review of 361 women at risk of carrying an affected fetus, 311 (86.2%) opted for the non-invasive approach using CTR and/or placenta. The cost saving of this non-invasive approach was relatively small (HK$ 2,651) in comparison to the cost of the whole prenatal screening program. On the other hand, the non-invasive approach was more expensive than the direct invasive approach for low MCV couples, as well as couples discordant for α-thalassemia and β-thalassemia. ages. These results support the adoption of non-invasive approach in which routine invasive test or karyotyping is no longer performed. A total of 106 at-risk pregnancies and normal controls were prospectively studied using three-dimensional ultrasonography. Placental volume (PV) at 11-14 weeks, and PV/CRL quotient at 9-14 weeks’ gestation of affected pregnancies were significantly greater than unaffected pregnancies (P<0.05). Using a cut-off point of 1.2ml/mm for PV/CRL quotient to predict an affected pregnancy, the sensitivity, and specificity was 96.2%, and 100.0% respectively. / published_or_final_version / Obstetrics and Gynaecology / Master / Doctor of Medicine

There has been an increase in rates of chromosomal abnormalities in newborns as a result of reproductive aging. For the past decades, a lot of effort has been placed on identifying pregnancies at risk of genetic defects. Conventional prenatal genetic diagnosis is achieved by invasive procedures that have been associated with an increased risk of pregnancy loss. This has led the researchers to explore the use of non-/minimally invasive techniques for prenatal diagnosis. Trophoblasts are known to be shed from regressing chorionic villi into the lower uterine pole of pregnant women during the first trimester. These cells are trapped within cervical mucus, which can be retrieved with a cytobrush. By using human leukocyte antigen G (HLA-G) and cytokeratin-7 (CK7) as trophoblast markers, this study aims to investigate the possibility of isolating individual fetal trophoblast from transcervical samples for genetic diagnosis. 195 healthy pregnant women requesting for legal termination of pregnancy (TOP) were recruited in this study. Transcervical cells were collected from them with the use of a cytobrush before TOP. HLA-G+ or CK7+ cells were then isolated by a combination of mucolytic action, fluorescent immunohistochemistry, and micromanipulation. The origin of these cells was subsequently investigated by either fluorescent in situ hybridization (FISH) or allelic profiling by quantitative fluorescent polymerase chain reaction (QF-PCR) based on chromosome 16, chromosome X, amelogenin gene and sex determining region Y (SRY) gene. This study first demonstrated the presence of fetal cells in transcervical samples based on the detection of chromosome Y signal by ordinary PCR. Cells expressing HLA-G and CK7 were also identified among transcervical cells. Immunopositive cells were isolated by micromanipulation under fluorescent microscopy. One isolated cell expressing CK7 was shown to inherit paternal allele at a locus on chromosome 16, suggesting the possible fetal origin of this cell. However, this study was still hampered by a number of technical factors. Further optimization of the protocol is required before transcervical trophoblasts can be retrieved in a reliable manner. / published_or_final_version / Obstetrics and Gynaecology / Master / Master of Philosophy

published_or_final_version / abstract / toc / Physiology / Master / Master of Philosophy

The effects of maternal dietary glucose restriction on reproductive performance were investigated by feeding pregnant rats isocaloric diets containing graded levels of dietary glucose (0, 12, 24 and 60%) during pregnancy and during pregnancy and lactation, and by measuring the effects of glucose restriction on (1) maternal, fetal and neonatal metabolism, on (2) growth and composition of the mammary glands and placentas, and (3) on milk composition. Carbohydrate restriction induced maternal metabolic adaptations that were proportional to the severity of the glucose restriction. Placental growth and composition as well as mammary gland composition were not affected by dietary glucose restriction, whereas fetal growth and development and milk composition were significantly impaired when glucose was limited in the maternal diet. This suggests that the effects of dietary glucose on the fetus and on milk composition were not mediated by changes in placenta and mammary gland DNA, protein or glycogen concentrations. Complete dietary glucose restriction significantly depressed fetal liver, lung and heart glycogen concentrations; repletion of the maternal diets with 12 and 24% glucose restored cardiac glycogen to normal but not fetal lung glycogen and liver glycogen. Pups born to dams fed a glucose-free diet failed to survive longer than 24 h postpartum and that was associated with the low levels of tissue glycogen at birth in these pups. At birth, lung and liver glycogen concentration of pups of the 12 and 24% glucose diets was similar to pups of the control diet despite the fact that these reserves were depressed in utero; and these pups efficiently corrected the transient hypoglycemia observed following parturition. The effects of glucose restriction on fetal liver glycogen were not reflected by similar changes in fetal plasma insulin, glucagon and glucose levels or in glycogen synthase and phosphorylase activities. Maternal dietary glucose was an important determinant

The study objectives were to: (1) assess fetal exposure to oxidative stress by measuring amniotic fluid concentrations of nitric oxide (NO), thiobarbituric acid--reactive substances (TBARS), and ferric reducing antioxidant power (FRAP) and (2) establish whether these concentrations were associated with infant birth weight, gestational age, or oxidative stress-related conditions arising during pregnancy. Frozen amniotic fluid samples were obtained from 654 mothers undergoing amniocentesis for genetic testing during second trimester in Montreal, QC, Canada. Maternal and neonatal characteristics were collected from medical charts and questionnaires and exclusion criteria were applied. ANOVAs and multivariate regression analyses showed that NO, which differed among pre-term, term, and post-term groups, was a positive predictor of gestational age. TBARS were highly correlated with sample storage and were not associated with pregnancy outcome parameters. FRAP positively predicted gender-corrected birth-weight-for-gestational-age. Our study shows that markers of oxidative and nitrative stress in-utero are associated with pregnancy outcomes.

The study objective was to determine whether early 2 nd trimester amniotic fluid (AF) erythropoietin (EPO) was associated with and predictive of (a) development of maternal gestational diabetes (GDM) and (b) the infant outcome parameters of (i) gestational age at birth (GAAB) assessed exclusively among spontaneous vaginal deliveries or (ii) birth weight (measured in grams and percentiles). Enzyme-linked-immunosorbent assay was used to determine the EPO concentration of 170 biobanked AF samples. Student's t-test revealed no difference between GDM and non-GDM subjects. AF EPO was not predictive of GAAB despite being significantly greater among preterm infants compared to post-term infants. In contrast, AF EPO was significantly higher among the smallest infants using both birth weight classification schemes. However, following inclusion of known covariates AF EPO was predictive of gram birth weight only. Early 2nd trimester AF EPO may emerge as a useful biomarker of fetal nutritional status and/or growth.

No description available.

The objectives were (1) to establish a profile of total fatty acids and cholesterol in amniotic fluid (AF) as well as (2) to determine possible associations between AT fatty acids (micromolar and relative proportion) with gestational age and birth weight. A total of 208 AF samples collected between 12 and 22 weeks of gestation during routine amniocentesis were analyzed using tandem column gas chromatography (GC). Smoking increased AF polyunsaturated fatty acid (PUFAs) levels while developmental stage and storage time decreased AF fatty acid quantities. AF trans fatty acids (TFAs) were negatively associated with both birth outcomes, whereas specific fatty acids including stearic acid (C18:0) and gondoic acid (C20:1n-9) were identified as negative predictors for gestational age and birth weight respectively. This study demonstrated novel relationships between fatty acids and fetal growth and gestational age in early midgestation AF, suggesting a possible role of AF fatty acids in predicting birth outcomes.

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