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Cultured human, ampullary, epithelial cells obtained from fertile women undergoing hysterectomy were evaluated for the support of human embryonic cleavage and growth in vitro. Twelve patients provided 23 embryos for co-culture with subcultured ampullary cells grown in T6 + 15% patient's serum and 18 embryos for growth in T6 + 15% patient's serum alone (controls). Of embryos co-cultured with ampullary cells, 78% cleaved to the compacted embryo stage and 69% cavitated as compared with 50 and 33% respectively for controls (P less than 0.01). Only 30% of co-cultured embryos reached the expanded blastocyst and 26% underwent hatching as compared with 28% for both stages in controls. At the 2 - 4- and 6 - 8-cell stages, 91 and 87% of co-cultured embryos showed an absence or slight fragmentation as compared with 72 and 61% respectively for embryos grown in medium alone (P less than 0.01). None of the co-cultured embryos showed unequal-sized blastomeres while 22% of controls showed unequal cleavage. Embryos grown with ampullary cells cleaved slightly faster than controls. Scanning electron micrographs showed that ampullary cells collected from co-cultures were all of the secretory type with several microvilli and apical protrusions. It is clear that subcultured human ampullary cells support human embryonic cleavage and yield a reasonable number of good quality embryos up to the cavitation stage. Development past the expanded blastocyst and hatching stages seems to involve another critical phase with its own specific requirements. [ABSTRACT FROM AUTHOR]

Objectives: To investigate multi-dose and timings of COVID-19 vaccines in preventing antenatal infection., Design: Prospective observational study investigating primary vaccinations, boosters, antenatal COVID-19 infections, neutralizing antibody (Nab) durability, and cross-reactivity to Delta and Omicron variants of concern (VOCs)., Results: Ninety-eight patients completed primary vaccination prepregnancy (29.6%) and antenatally (63.3%), 24.2% of whom had antenatal COVID-19, while 7.1% were unvaccinated (28.6% had antenatal COVID-19). None had severe COVID-19. Prepregnancy vaccination resulted in vaccination-to-infection delay of 23.3 weeks, which extended to 45.2 weeks with a booster, compared to 16.9 weeks following antenatal vaccination (P < 0.001). Infections occurred at 26.2 weeks gestation in women vaccinated prepregnancy compared to 36.2 weeks gestation in those vaccinated during pregnancy (P < 0.007). The risk of COVID-19 infection was higher without antenatal vaccination (hazard ratio [HR] 14.6, P = 0.05) and after prepregnancy vaccination without a booster (HR 10.4, P = 0.002). Antenatal vaccinations initially led to high Nab levels, with mild waning but subsequent rebound. Significant Nab enhancement occurred with a third-trimester booster. Maternal-neonatal Nab transfer was efficient (transfer ratio >1), and cross-reactivity to VOCs was observed., Conclusion: Completing vaccination during any trimester delays COVID-19 infection and maintains effective neutralizing activity throughout pregnancy, with robust cross-reactivity to VOCs and efficient maternal-neonatal transfer., Competing Interests: Declarations of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Competing Interests: Declaration of interests We declare no competing interests.

Introduction: Lactogenesis II (LaII) failure can be prevented in at-risk mothers with simple proactive interventions. In a randomised trial, we investigated the efficacy of early and regular breast milk expression in establishing LaII, using an electric double-breast pump., Methods: Mothers with uncomplicated singleton deliveries were randomised to intervention (n = 31) or control (n = 29) groups. The former commenced breast milk expression with an electric pump within one hour of delivery and maintained regular expression with direct breastfeeding. Control mothers directly breastfed without regular pump expression. Expressed milk volumes were analysed for citrate, lactose, sodium and protein., Results: Median time of LaII was Day 3 (interquartile range [IQR] 1 day) with intervention and on Day 4 (IQR 1 day) among controls (p = 0.03). Biochemical steady-state concentrations were achieved around early Day 4 (sodium, total protein) and Days 4-5 (citrate, lactose). Sodium, protein and lactose levels were similar in both groups over seven days, at 5.80 mM, 0.68 mM and -13.38 mM, respectively. Mean daily milk volume with intervention was 73.9 mL on Day 3 and 225.2 mL on Day 7, greater than controls (25.4 mL on Day 3 and 69.2 mL on Day 7; p < 0.2). Mean infant weights were similar on Day 8 at 3,477 g with intervention and 3,479 g among controls., Conclusion: LaII is established by postnatal Day 3 with early initiation of regular breast milk expression, a useful intervention for mothers at risk of early-onset breastfeeding failure., (Copyright: © Singapore Medical Association.)

Introduction: In 2006, Singapore adopted the universal hepatitis B immunoglobulin (HBIg) policy. Since then, all infants of hepatitis B surface antigen (HBsAg)-positive mothers receive HBIg, irrespective of maternal hepatitis B e antigen (HBeAg) status. However, the benefits of HBIg for infants of HBeAg-negative mothers are unclear. We compared the vertical transmission rates among children of HBeAg-negative mothers who were given HBIg versus a retrospective cohort who were not given HBIg, to determine its protective effect., Methods: This observational study involved pregnant HBsAg-positive women seen at National University Hospital, Singapore, between June 2009 and December 2013. If the infants of these mothers completed the recommended vaccination schedule, they were recruited into the study, along with their older siblings. Serological testing for the children was performed three months after completion of the last dose of vaccine, and hepatitis B virus (HBV) surface gene sequencing was carried out if HBV DNA was detected., Results: A total of 111 infants and 47 siblings were recruited. 2 (1.5%) children were found to have vertical transmission despite receiving HBIg, while no incidences of vertical transmission were found among the historical controls who did not receive HBIg (p = 1.00)., Conclusion: The overall effectiveness of the hepatitis B vaccination programme for children of HBsAg-positive mothers was high, regardless of HBIg administration. The addition of HBIg did not appear to confer additional benefits, in terms of vertical transmission rate, among infants born to HBeAg-negative mothers., (Copyright: © Singapore Medical Association)

Hepatitis B virus (HBV) infection is usually vertically transmitted from the mother to child during birth in Asian countries. Despite immunization, immunoprophylaxis failure is well-documented. The aim of the study was to study immunoprophylaxis failure rate in the cohort of infants delivered by chronic HBV-infected mothers and to determine risk factors for failure. This was an observational study involving chronic hepatitis B infected mothers seen at a tertiary care center in Singapore between June 2009 and December 2013. Infants born to these mothers were recruited after they had completed the recommended vaccination schedule. Serological testing for the children was performed 3 months after completion of the last dose of vaccine. HBV surface gene sequencing was carried out if HBV DNA was detectable in the children. Among the 161 mothers enrolled, most were HBeAg negative. HBeAg positive mothers were younger and had a significantly higher viral load (6.5 log) as compared to HBeAg negative mothers (1.35 log) (P < 0.001). Four children (2.6%) were found to have immunoprophylaxis failure. Two occurred in children delivered by mothers with extremely high viral load of more than 5 × 10(7)  IU/ml. HBV surface gene mutations were detected in most children (3 out of 4) with immunoprophylaxis failure. The overall effectiveness of the hepatitis B vaccination program was high. High maternal viral load and presence of surface gene mutants may be potential contributors., (© 2015 Wiley Periodicals, Inc.)

Introduction: Breast milk fatty acids play a major role in infant development. However, no data have compared the breast milk composition of different ethnic groups living in the same environment. We aimed to (i) investigate breast milk fatty acid composition of three ethnic groups in Singapore and (ii) determine dietary fatty acid patterns in these groups and any association with breast milk fatty acid composition., Materials and Methods: This was a prospective study conducted at a tertiary hospital in Singapore. Healthy pregnant women with the intention to breastfeed were recruited. Diet profile was studied using a standard validated 3-day food diary. Breast milk was collected from mothers at 1 to 2 weeks and 6 to 8 weeks postnatally. Agilent gas chromatograph (6870N) equipped with a mass spectrometer (5975) and an automatic liquid sampler (ALS) system with a split mode was used for analysis., Results: Seventy-two breast milk samples were obtained from 52 subjects. Analysis showed that breast milk ETA (Eicosatetraenoic acid) and ETA:EA (Eicosatrienoic acid) ratio were significantly different among the races (P = 0.031 and P = 0.020), with ETA being the highest among Indians and the lowest among Malays. Docosahexaenoic acid was significantly higher among Chinese compared to Indians and Malays. No difference was demonstrated in n3 and n6 levels in the food diet analysis among the 3 ethnic groups., Conclusions: Differences exist in breast milk fatty acid composition in different ethnic groups in the same region, although no difference was demonstrated in the diet analysis. Factors other than maternal diet may play a role in breast milk fatty acid composition.

Introduction: We aimed to develop a rapid quantitative-fluorescence polymerase chain reaction (QF-PCR) to detect common foetal aneuploidies in the Singapore population within 48 hours of sample collection in order to alleviate parental anxiety., Methods: DNA from 1,000 foetal samples (978 amniotic fluids, 14 chorion villi and eight foetal blood samples) was analysed using a QF-PCR of 19 microsatellite markers located on chromosomes 13, 18, 21, X and Y. A total of 523 samples were archived before the QF-PCR analysis (archived), while QF-PCR was performed and the results obtained within 48 hours of sample collection in the remaining 477 samples (live). The results were confirmed with their respective karyotypes., Results: In total, 47 autosomal trisomies (T) were found: 30 among the archived (three T13, 12 T18, 15 T21) and 17 among the live (four T18, 13 T21) samples. The QF-PCR results were verified with their respective karyotypes. We achieved 100 percent sensitivity (lower 95 percent confidence interval [CI], 92.8 percent) and specificity (lower 95 percent CI, 99.5 percent), and the time taken from sample collection to the obtaining of results for the 477 live samples was less than 48 hours., Conclusion: Prenatal diagnostic results of common chromosomal abnormalities can be released within 48 hours of sample collection using QF-PCR. Parental anxiety is alleviated and clinical management is enhanced with this short waiting time.

Objective: Prenatal diagnosis of alpha-thalassaemia requires invasive testing associated with a risk of miscarriage. Cell-free foetal DNA in maternal plasma presents an alternative source of foetal genetic material for noninvasive prenatal diagnosis. We aimed to exclude HbBart's noninvasively by detection of unaffected paternal alleles in maternal plasma using quantitative fluorescence PCR (QF-PCR)., Method: Microsatellite markers (16PTEL05, 16PTEL06) within the breakpoint regions of -(SEA), -(FIL) and -(THAI) deletions were analysed using QF-PCR of maternal plasma from 30 families. In this blinded study, genotypes were confirmed using conventional PCR. Maternal plasma from two known cases of HbBart's were also analysed., Results: HbBart's was excluded in 10 out of 30 (33.3%, 95% CI, 17.3-52.8%) mothers by identifying the presence of nondeleted paternally inherited fetal alleles; either only 16PTEL05 (n = 1) or only 16PTEL06 (n = 4), or both (n = 5), and confirmed through direct analysis of fetal DNA. Paternally inherited foetal alleles of 16PTEL05 and 16PTEL06 were not detected in maternal plasma of the two known HbBarts cases. False negatives were excluded with the detection of paternally inherited fetal control marker, D21S1270 in maternal plasma., Conclusion: We show proof-of-principle that such a test can accurately exclude HbBart's in the foetus by identifying the nondeleted paternally inherited fetal alleles in maternal plasma in one out of three pregnancies, avoiding invasive testing in these pregnancies., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

Objective: Prevention of postpartum haemorrhage is essential in the pursuit of improved health care for women. However, limited literature is available for comparing the use of oxytocin agonist carbetocin with syntometrine in women undergoing vaginal deliveries. We aimed to compare intramuscular carbetocin with intramuscular syntometrine for the routine prevention of postpartum haemorrhage in women who deliver vaginally., Design: Prospective double-blind randomised controlled trial., Setting: Tertiary referral centre., Population: Pregnant women with no contraindication for vaginal delivery recruited from January 2005 to April 2008., Methods: Participants were randomised to receive either syntometrine or carbetocin during the third stage of labour., Main Outcome Measures: Primary outcome measure was postpartum haemorrhage requiring additional uterotonics. Secondary outcome measures were the incidence of postpartum haemorrhage (> or =500 ml), severe postpartum haemorrhage (> or =1000 ml) and adverse effects profile., Results: Women in the carbetocin group (13.5%) and in the syntometrine group (16.8%) had postpartum haemorrhage requiring additional uterotonics (P = 0.384). 1.6% of women in each group had postpartum haemorrhage (P = 1.0) and the estimated blood loss during the third stage of labour was similar between the two groups (P = 0.294). Women who had syntometrine were four times more likely to experience nausea (RR = 4.2; 95% CI 2.2-7.8) and vomiting (RR = 4.3; 95% CI 1.9-9.5) compared with women who had carbetocin. Tremor, sweating, retching and uterine pain were also more likely in the syntometrine group compared with the carbetocin group (P < 0.05)., Conclusions: Carbetocin has an efficacy similar to syntometrine for prevention of postpartum haemorrhage, but is associated with less adverse effects.

Objective: To investigate whether antenatal breast feeding education alone or postnatal lactation support alone improves rates of exclusive breast feeding compared with routine hospital care., Design: Randomised controlled trial., Setting: A tertiary hospital in Singapore., Participants: 450 women with uncomplicated pregnancies., Main Outcome Measures: Primary outcomes were rates of exclusive breast feeding at discharge from hospital and two weeks, six weeks, three months, and six months after delivery. Secondary outcomes were rates of any breast feeding., Results: Compared with women who received routine care, women in the postnatal support group were more likely to breastfeed exclusively at two weeks (relative risk 1.82, 95% confidence interval 1.14 to 2.90), six weeks (1.85, 1.11 to 3.09), three months (1.87, 1.03 to 3.41), and six months (2.12, 1.03 to 4.37) postnatally. Women receiving antenatal education were more likely to breast feed exclusively at six weeks (1.73, 1.04 to 2.90), three months (1.92, 1.07 to 3.48), and six months (2.16, 1.05 to 4.43) postnatally. The numbers needed to treat to achieve one woman exclusively breast feeding at six months were 11 (6 to 80) for postnatal support and 10 (6 to 60) for antenatal education. Women who received postnatal support were more likely to exclusively or predominantly breast feed two weeks after delivery compared with women who received antenatal education (1.53, 1.01 to 2.31). The rate of any breastfeeding six weeks after delivery was also higher in the postnatal support group compared with women who received routine care (1.16, 1.02 to 1.31)., Conclusions: Antenatal breast feeding education and postnatal lactation support, as single interventions based in hospital both significantly improve rates of exclusive breast feeding up to six months after delivery. Postnatal support was marginally more effective than antenatal education., Trial Registration: Clinical Trials NCT00270920 [ClinicalTrials.gov].

Cell membrane proteome analysis is limited by inherent membrane hydrophobicity. Conventional membrane protein extraction techniques use detergents, chaotropes and organic acids that require sample clean-up or pH adjustment, and are associated with significant sample loss. We extracted membrane proteins from red blood cells (RBCs) using methanol (MeOH), trifluoroethanol (TFE) and urea, and identified membrane proteins using 2-D LC coupled with MALDI-TOF/TOF-MS. We show that organic solvents MeOH- and TFE-based methods have membrane protein analysis efficiencies comparable to urea, and are complementary for the recovery of both hydrophilic and hydrophobic peptides. The mean grand average of hydropathicity (GRAVY) value of identified peptides from the TFE-based method (-0.107) was significantly higher than that of the MeOH-based method (-0.465) (p<0.001). Sequential and adjunctive use of the organic solvents MeOH and TFE increases the number of proteins identified, and the confidence of their identification. We show that this strategy is effective for shotgun membrane proteome analysis.

Background: We sought to develop a rapid prenatal diagnostic test for simultaneous detection of HbBarts hydrops fetalis and exclusion of maternal contamination., Methods: We developed a multiplex quantitative fluorescent PCR (QF-PCR) test that detects the presence/ absence of 2 microsatellite markers (16PTEL05/16PTEL06) located within breakpoints of the Southeast Asia ((-SEA)) deletion. HbBarts hydrops fetalis ((-SEA/-SEA)) is diagnosed by absence of both markers, and maternal contamination of fetal DNA is excluded by absence of noninherited maternal alleles. Fetal and parental DNA samples from 50 families were analyzed in a blinded clinical validation study, and QF-PCR results were compared with their respective molecular genotypes., Results: The multiplex QF-PCR results included correct diagnoses of HbBarts hydrops fetalis in 11 of the fetuses tested, correct verification as unaffected in 20 fetuses, and correct identification as either carriers (alphaalpha/(-SEA)) or unaffected homozygotes in 18. Misidentification as unaffected occurred for 1 carrier. Sensitivity for diagnosis of HbBarts hydrops fetalis was 100% [lower 95% confidence interval, 76.2%], and specificity was 100% (lower 95% confidence interval, 92.6%). None of the samples tested showed any traces of noninherited maternal alleles; thus false-positives because of maternal contamination were eliminated., Conclusions: In this QF-PCR method, detection of maternally and paternally inherited fetal alleles allowed diagnosis of the double-deletion syndrome, and the ability to differentiate between these alleles allowed simultaneous exclusion of maternal contamination of the fetal genetic material. This novel strategy using cell-free fetal DNA in maternal plasma could form the basis for noninvasive testing for HbBarts hydrops fetalis.

Background: Mast cells are well established effectors of IgE-triggered allergic reactions and immune responses to parasitic infections. Recent studies indicate that mast cells may play roles in adaptive and innate immunity, suggesting an innovative view of the regulation of immune responses. Here, we profiled the transcriptome of human mast cells sensitized with IgE alone, or stimulated by FcepsilonRI aggregation., Results: Our data show that among 8,793 genes examined, 559 genes are differentially regulated in stimulated mast cells when compared with resting/unstimulated mast cells. The major functional categories of upregulated genes include cytokines, chemokines, and other genes involved in innate and adaptive immune-responses. We observed the increased expression of over 63 gene-transcripts following IgE-sensitization alone. Our data was validated using Real-Time-PCR; ELISA and western blot. We confirmed that IgE alone does not trigger mast cell-immediate responses, such as calcium signals, degranulation or protein-phosphorylation., Conclusion: This report represents a substantial advance in our understanding of the genome wide effects triggered by "passive sensitization" or active stimulation of human mast cells, supporting mast cells' potential involvement in a wide range of inflammatory responses.

Objectives: To evaluate the fetal cardiac time intervals from the longitudinal analysis of noninvasive fetal electrocardiography (fECG) in normal pregnancies., Methods: One hundred singleton pregnancies were examined in this longitudinal study. Cardiac time intervals were derived from fetal electrocardiograms obtained noninvasively using three electrodes placed on the maternal abdomen. The variables measured included the durations of the P wave, PR interval, QRS complex, QT interval and T wave., Results: Success rates for detecting the P, QRS and T waves were 74.6, 91.0 and 79.3%, respectively. Cardiac time intervals were significantly influenced by fetal age. The mean P-wave duration increased from 43.9 (18--22 weeks) to 52.9 ms (>/=37 weeks) (p < 0.001). PR intervals were 102.1 and 110.1 ms, for fetuses at 18 to 22 and >/=37 weeks (p < 0.001), respectively. QRS intervals were 47.2 and 52.6 ms (p < 0.001), while QT intervals were 224.0 and 242.7 ms (p < 0.001), at 18 to 22 and >/=37 weeks respectively. From 18 to 22 weeks to >/=37 weeks, QT(c) values increased from 343.8 to 367.7 ms (p < 0.001), while T-wave durations increased from 123.8 to 152.4 ms (p < 0.001)., Conclusions: Serial noninvasive fECG of normal fetuses from 18 to 41 weeks of gestation show good success rates of fECG detection. Cardiac time intervals generally increased with increasing gestational age., (Copyright (c) 2005 John Wiley & Sons, Ltd.)

The 75 and 50g OGTT (Oral Glucose Tolerance Test) responses were studied in 86 normal healthy pregnant women (mean age 28.7 +/- 0.4 (SEM) years) at 28 and 32 weeks respectively. Of these were 50 Chinese, 20 Malays and 16 Indians. Mean glucose responses at fasting, 1 and 2h post glucose load were 78.3 +/- 0.7, 132.2 +/- 2.8 and 116.2 +/- 2.1 mg/dl respectively for the 75g OGTT and 78.5 +/- 0.7, 130.5 +/- 2.5 and 106.7 +/- 1.8 mg/dl respectively for the 50g OGTT. Except for the 2h responses, corresponding responses between both OGTTs were not significantly different. All races showed a similar OGTT response. Using a set criteria for diagnosis of abnormals resulted in gross inconsistency in the number of abnormals diagnosed for both OGTTs. However, the use of 95th percentile shows a closer agreement in the diagnosis of abnormal cases for both OGTTs. Also, the 2h OGTT response for the 75g OGTT is higher than that of WHO's criteria for impaired glucose tolerance. This emphasizes the need to establish our own reference range.

Serum fructosamine levels in women at pregnancy (28 weeks' gestation) were determined. The women (n = 99) also participated in an oral glucose tolerance test (OGTT) with a 75g liquid glucose load, to determine their glucose tolerance. At 32 weeks' gestation, seventy-nine of them repeated the oral glucose tolerance test and fructosamine measurement. Results showed that fructosamine levels in pregnant women with normal glucose tolerance (2.20 +/- 0.19 mmol/l, n = 76), were not statistically different from those with glucose intolerance (gestational diabetes: 2.19 +/- 0.22 mmol/l, n = 23) at 28 weeks' and also 32 weeks' gestation. However, serum fructosamine levels in pregnant women were lower than those in non-pregnant subjects. Serum fructosamine measurement is not a sufficiently sensitive test for diagnosis of gestational diabetes.

24-hour short term culture of trophoblastic villi provided rapid and accurate karyotypes in spontaneous first trimester abortions. Good quality metaphases were obtained in 60.0% of villous cultures if these were set up within 8 hours of uterine evacuation, in contrast to the 10.3% success rate with intervals longer than 8 hours. 21% (4 of 19) of first trimester pregnancy losses were chromosomally abnormal, 3 of which were examples of autosomal trisomy; in 2 cases, the mothers were more than 37 years of age. The results of successful karyotyping reported here compare well with previous reports of long term culture of minced fetal material. Further, the risks of infected cultures, maternal contamination and pseudomosaicism are minimal with the short-term culture technique described here. Chorionic villi isolated from spontaneously aborted material are a simple and practicable source of fetal material for rapid and accurate cytogenic diagnosis in early spontaneous abortions.

With national policies aimed at reaching zero population growth in the shortest possible time, many countries have introduced restrictive legislation and disincentive programs in an attempt to decrease family size norms to 2 or 3 children. As a result, younger women of lower parity are being sterilized, with consequences that will be seen in the years to come. Although female sterilization is usually associated with minimal complications and side effects and is highly effective without continuing motivation or promotion, it has the disadvantage of causing permanent, essentially irreversible sterility. Therefore, it will not be readily acceptable to a large portion of the population. For many women, alternatives must be sought. Reversible methods of female or male sterilization, longacting systemic contraceptives, longacting implants, and immunization against implantation or sperm antigens are potential alternatives, but all are still in experimental stages of development. Intrauterine devices and injectables are the 2 most effective alternatives now available. The use of intrauterine devices with abortion as a backup in case of contraceptive failure ranks high as an alternative to female sterilization.

Epithelial cells from the ampulla of healthy oviducts from 16 women aged 33-41 years and at different phases of their menstrual cycles were used to establish primary cultures and the continuation of a cell line. The morphology and behaviour of these cells in vitro were evaluated using Nomarski's inverted optics, scanning and transmission electron microscopy. Cells from all patients produced confluent monolayers in 6-7 days with no significant relationship of cell growth with stage of cycle. Fourteen primary cultures were of the epithelioid type while two showed mixed epithelioid and fibroblast-like growth. Two distinct cell types (ciliated and secretory) were observed in primary culture. Secretory cells showed several microvilli of different lengths and distribution. Secretory cells predominated over ciliated cells in all patients, but maximum ciliation occurred around the time of ovulation. Structural features of the cells in vitro were remarkably similar to those described in vivo. Ampullary cells could be maintained in vitro through four to six passages with 3-4 days of growth between passages. Sub-cultured cells were all secretory and were of two types (I and II) based on ultrastructure. Secretory vesicles containing electron-dense material and lipids were observed in these cells. The method described allows for the use of ampullary cells as feeder layers for IVF and support of cleaving human embryos and the evaluation of the biochemical events surrounding fertilization and ectopic pregnancies.