Allotransplantation of transgenic mouse ovaries expressing enhanced green fluorescent protein under the control of the murine phosphoglycerate kinase 1 promoter.

The aim of this study was to generate a transgenic mouse that ubiquitously expressed enhanced green fluorescent protein (EGFP) under the control of the murine phosphoglycerate kinase 1 promoter by allotransplantation of transgenic mouse ovaries. The EGFP transgenic mice expressed green fluorescence in many organs, and the fluorescence was detected as early as the embryonic stage. Ovaries from the EGFP transgenic mice were allotransplanted into recipients and these mice were mated with normal male mice. Histological sections of EGFP-allotransplanted ovaries from the recipient mice showed the well development and formation at follicles and corpora lutea. The green fluorescence was clearly detectable at the allotransplanted section of the ovaries, which had fused with the normal ovary. The average size of the first litter from these mice was 6.8 ± 1.2 pups per recipient, and 17.8% of the pups expressed EGFP. These results demonstrated that allotransplantation of transgenic ovaries can restore a normal reproductive lifespan and can be used to generate a ubiquitously expressing EGFP animal model.

Vitamin D deficiency and supplementation during pregnancy.

OBJECTIVE: Vitamin D is essential for skeletal health and prolonged deficiency results in infantile rickets and adult osteomalacia. The aim of this study is to determine the vitamin D status in pregnancy and to evaluate the effects of daily and of single-dose vitamin D supplementation. DESIGN: A prospective randomized study at St Mary's Hospital London. PATIENTS: A total of 180 women (Indian Asian, Middle Eastern, Black and Caucasian) were recruited at 27 weeks gestation and randomized into three treatment groups: a single oral dose of 200,000 IU vitamin D, a daily supplement of 800 IU vitamin D from 27 weeks until delivery and a no treatment group. MEASUREMENTS: Vitamin D (25-hydroxyvitamin D), PTH and corrected calcium levels in mothers at 27 weeks and at delivery and cord 25-hydroxyvitamin D and corrected calcium levels. RESULTS: The final maternal 25-hydroxyvitamin D levels were significantly higher in the supplemented group [daily dose (median) 42 (IQR 31-76) nmol/l, stat dose (median) 34 (IQR 30-46) nmol/l vs. median 27 (IQR 27-39) nmol/l in the no treatment; P < 0.0001] and significantly fewer women with secondary hyperparathyroidism in the supplemented group (10% in daily dose vs. 12% in stat dose vs. 27% in the no treatment; P < 0.05). Cord 25-hydroxyvitamin D levels were significantly higher with supplementation [daily dose median 26 (IQR 17-45) nmol/l, stat dose median 25 (IQR 18-34) nmol/l vs. median 17 (IQR 14-22) nmol/l in no treatment; P = 0.001]. CONCLUSION: Single or daily dose improved 25-hydroxyvitamin D levels significantly. However, even with supplementation, only a small percentage of women and babies were vitamin D sufficient. Further research is required to determine the optimal timing and dosing of vitamin D in pregnancy.

Outcome of pregnancy in sickle cell disease patients attending a combined obstetric and haematology clinic.

This study aims to determine the pregnancy outcomes in women attending the combined obstetric sickle cell clinic at King's College Hospital, London from June 2000 to July 2006. There were 71 pregnancies in 65 women with sickle cell disease. Sickle crisis requiring admission occurred in 47% of the antenatal patients. The first admission occurred most frequently in the third trimester (23 vs 6 and 5 admissions in the second and first trimester; p < 0.001). There were no maternal deaths; other complications include anaemia requiring blood transfusion (32%), proteinuric hypertension (9%), infections (28%) and emergency caesarean section (30%). There was one fetal demise due to abruption. Pre-term delivery before 34 weeks occurred in 8% and 24% before 37 weeks. A total of 18% of infants had reduced growth velocity with the measurements crossing below the 10th centile.

Prediction of pre-eclampsia by uterine artery Doppler imaging: relationship to gestational age at delivery and small-for-gestational age.

OBJECTIVES: To determine the relationship between pre-eclampsia, small-for-gestational age (SGA) and gestational age at delivery, and the effect of this relationship on the prediction of pre-eclampsia by uterine artery Doppler imaging. METHODS: This was a multicenter prospective Doppler study of the uterine artery at 22-24 weeks of gestation in unselected women with singleton pregnancies. RESULTS: In the 30,639 pregnancies examined, the median uterine artery pulsatility index (PI) was 1.0 and the 95(th) centile was 1.58. In 614 (2%) cases the woman developed pre-eclampsia and in this group there was an inverse significant association between the gestational age at delivery and prevalence of SGA (r = - 0.99, P < 0.0001), and between the gestational at delivery and mean uterine artery PI (r = - 0.51, P < 0.0001) and prevalence of mean uterine artery PI above the 95(th) centile (r = - 0.99, P < 0.0001). The mean uterine artery PI was above the 95(th) centile in 77.2% of women who developed pre-eclampsia requiring delivery before 34 weeks, in 35.9% of those delivering at 34-37 weeks and in 21.9% of those delivering after 37 weeks. The respective percentages were 82.3%, 46.9% and 28.8% for those with pre-eclampsia and SGA, and 43.8%, 21.2% and 8.4% for those with SGA but without pre-eclampsia. CONCLUSIONS: Pre-eclampsia requiring early delivery is more likely to be associated with SGA than less severe pre-eclampsia in women who deliver at term. Doppler ultrasound assessment of the uterine arteries is more effective in identifying pre-eclampsia requiring preterm than term delivery.

Prediction of pre-eclampsia by a combination of maternal history, uterine artery Doppler and mean arterial pressure.

OBJECTIVES: To determine the value of combined screening for pre-eclampsia by maternal history, and mid-trimester uterine artery (UtA) Doppler imaging and maternal blood pressure. METHODS: In 3529 singleton pregnancies attending for routine care at 22-24 weeks' gestation we recorded maternal variables, and made UtA Doppler and mean arterial pressure (MAP) measurements. Multiple regression analysis was used to determine the significant predictors of pre-eclampsia, gestational hypertension and small-for-gestational age (SGA) among maternal characteristics, UtA pulsatility index (PI) and MAP. RESULTS: Complete pregnancy outcomes were available in 3359/3529 (95.2%) cases. Pre-eclampsia developed in 101 (3.0%) pregnancies, including 23 (0.7%) in which delivery was before 34 weeks (early pre-eclampsia) and 78 (2.3%) with delivery at 34 weeks or more (late pre-eclampsia); 74 (2.2%) developed gestational hypertension, 366 (10.9%) delivered SGA newborns with no hypertensive disorders, and 2806 (83.8%) were unaffected by pre-eclampsia, gestational hypertension or SGA. Multiple regression analysis demonstrated that maternal characteristics, UtA-PI and MAP provided a significant independent contribution in the prediction of pre-eclampsia, gestational hypertension and SGA. For a false-positive rate of 10%, the estimated detection rates of early and late pre-eclampsia were 100% and 56.4%, respectively. CONCLUSIONS: The combination of maternal demographic characteristics, and UtA Doppler and maternal blood pressure measurements is an effective screening tool for the prediction of pre-eclampsia.

Cerebrospinal fluid cytokines in enterovirus 71 brain stem encephalitis and echovirus meningitis infections of varying severity.

Taiwan has experienced several outbreaks of enterovirus 71 (EV71) infections since 1998. This study examined the quantitative relationship between specific cytokines in the cerebrospinal fluid (CSF) and the severity of EV71 brain stem encephalitis (BE), and investigated whether the CSF cytokine response differed from that to uncomplicated echovirus meningitis (EM). The study included 57 children with EV71 BE, of whom 24 had isolated BE, 24 had autonomic nervous system (ANS) dysregulation, and nine had pulmonary oedema (PE), and 15 children with EM. All were confirmed by virus culture. Mean CSF glucose, total protein and lactate levels were increased significantly in association with the severity of EV71 BE. The mean CSF concentration of interleukin (IL)-1beta in children with EV71 PE was significantly higher than in those with isolated BE. IL-6 and interferon (IFN)-gamma levels were significantly higher for EV71 PE and ANS dysregulation than for isolated BE. In contrast, EM was associated with high levels of IL-1beta and low levels of IFN-gamma. Cytokines in the central nervous system, as well as in blood, appear to be involved in the pathogenesis of EV71 BE.

Remifentanil preconditioning confers delayed cardioprotection in the rat.

BACKGROUND: Preconditioning with remifentanil (RPC) provides immediate cardioprotection in rats via all three types of opioid (OP) receptor. This study sought to investigate whether remifentanil also confers delayed cardioprotection via OP receptors. METHODS: Male rats received preconditioning either by ischaemia (IPC; 5 min occlusion, 5 min reperfusion x 3) or with remifentanil (RPC; 1, 5, 10, and 20 microg kg(-1) min(-1), 20 min infusion). After 24 h, all animals were subjected to 30 min occlusion of the left coronary artery and 2 h of reperfusion. Subsequently, the time-course effect of RPC (10 microg kg(-1) min(-1), 20 min infusion) was determined at 12, 16, 24, 32, 36, and 48 h intervals, using the same experimental procedure. The effect of RPC (10 microg kg(-1) min(-1), 20 min infusion) and IPC in the presence of selective OP receptor antagonists was evaluated at the 24 h interval. Infarct size (IS), as a percentage of the area at risk (AAR), was determined. RESULTS: Pre-treatment with remifentanil at 1, 5, 10, and 20 microg kg(-1) min(-1) significantly reduced the IS/AAR at 24 h with the maximum effect at 10 microg kg(-1) min(-1). Remifentanil at 10 microg kg(-1) min(-1) significantly reduced the IS at 12 h [32.5 (sd 9.1)%]; 16 h [26.1 (2.8)%]; 24 h [19.5 (5.0)%]; 32 h [31.2 (9.1)%]; and 36 h [36.4 (9.4)%] after drug administration. The maximal reduction in IS was seen at 24 h and the effect completely disappeared at 48 h [36.4 (9.4)%]. The protective effect of RPC was abolished or significantly attenuated by blockade of any of the three OP receptors with selective antagonists. CONCLUSIONS: Like IPC, remifentanil produces delayed cardioprotection in anaesthetized rats 12-36 h after administration. The protective effect is mediated via all three OP receptors.

A non-biotin polymerized horseradish-peroxidase method for the immunohistochemical diagnosis of canine distemper.

This report describes a modified non-biotin polymerized horseradish peroxidase (HRP) immunohistochemical method for the diagnosis of canine distemper virus (CDV) infection from formalin-fixed, paraffin wax-embedded tissues. This method confirmed infection in seven of eight (87.5%) suspected cases. Labelled CDV antigen was observed in the following sites: cerebrum, cerebellum, meninges, glial cells, neurons, vascular endothelium, periventricular areas and pericytes, and choroid plexus; grey and white matter and central canal of the spinal cord; renal pelvis and tubular epithelium, and urinary bladder epithelium; macrophages and lymphocytes in splenic white pulp and lymph nodes; skin epidermis; bronchiolar epithelium and alveolar macrophages; hepatic Kupffer cells, and gastric and intestinal mucosal epithelium; stratified squamous epithelium of the tongue and oesophagus. With the non-biotin HRP detection system, pretreatment by autoclaving followed by microwave heating gave better labelling results than did microwave pretreatment alone. No obvious difference was noted between the labelling results produced by the non-biotin HRP detection system and the Super Sensitive Link-Label IHC detection system.

Mitotic delay and chromosomal aberrations induced by X rays in synchronized Chinese hamster cells in vitro.

Synchronized Chinese hamster cell populations in vitro were X-irradiated at different stages of the cell cycle and examined at the next metaphase. Mitotic delay varied according to the stage of the cell cycle at the time of irradiation. It was greatest for cells irradiated in middle to late S, moderate for cells irradiated in G2, and least for cells irradiated in G1. It increased approximately linearly with dose (0.7 hr./100 rad for S cells and 0.3 hr./100 rad for G1 cells). Chromosomal aberration rates also depended on the stage of the cell cycle at which the cells were irradiated. Chromatid aberrations were induced mostly during G2 and S (more in early S than late S), while chromosome aberrations were induced mainly in G1. Total breaks after 500 or 750 R showed a minimum frequency in late S and a maximum frequency in G2. This form of response correlates, but only in a general qualitative way, with the dependence of survival on cell cycle stage previously reported. Mitotic delay has a dependence on cell cycle stage quite different from either chromosomal aberrations or survival.

Attitudes towards cancer survivors: a small survey.

INTRODUCTION: The National Cancer Survivors Day Foundation defines a cancer "survivor" as anyone living with a history of cancer--from the moment of diagnosis through the remainder of life. Little is known about the size and make-up of this population or about the medical care experience of and social implications for patients who have had a diagnosis of cancer in Singapore. An opportunistic survey was undertaken to understand how members of the public believe about this population. METHODS: A sample of the general public was undertaken during the "CancerVive" event in 2004. Questionnaires regarding employment as well as attitudes towards cancer and cancer survivorship were distributed. RESULTS: Members of the public held certain misconceptions about cancer survivors. They also have certain negative attitudes toward cancer survivors. Beliefs and attitudes about cancer are similar for cancer survivors and the general public. Although members of the public had positive attitudes towards working with cancer survivors, the majority felt that cancer survivors should not be given equal opportunities at work, by not employing cancer survivors if they were in the position to hire. CONCLUSION: Further research with larger and more representative samples needs to be undertaken to extend the understanding into cancer survivorship issues.

Second-trimester sex hormone-binding globulin and subsequent development of pre-eclampsia.

OBJECTIVE: To investigate whether maternal plasma sex hormone-binding globulin (SHBG) concentrations are reduced in women who subsequently develop pre-eclampsia. METHODS: This was a cross-sectional study, carried out at antenatal clinics in seven hospitals in and around London. Healthy women underwent uterine artery Doppler velocimetry as a screening method for pre-eclampsia at 22-24 weeks of gestation. The first group (408 women) had normal uterine artery Doppler waveforms (mean uterine artery pulsatility index (PI) below 1.6). The second group (274 women) had increased impedance to flow in the uterine arteries (mean PI above the 95th centile, 1.6). Maternal plasma SHBG concentrations were measured retrospectively using a competitive chemiluminescent immunoassay. Pre-eclampsia was as defined by the International Society for the Study of Hypertension in Pregnancy. RESULTS: Plasma SHBG concentrations in the 80 (11.7%) women who subsequently developed pre-eclampsia were significantly lower than in the 585 (85.8%) women with normal pregnancy outcomes (median 336, range 142-674 nmol/l vs. median 336, range 142-674 nmol/l, p = 0.001). There was a strong correlation between SHBG concentrations and body mass index (r =-0.232246, p < 0.0001). There were no significant differences in maternal plasma SHBG concentrations in women with abnormal uterine artery Doppler (n = 274) compared with controls (n = 408) (median 324, range 101-635 nmol/l vs. median 336, range 142-674 nmol/l, p = 0.09). CONCLUSION: Maternal plasma SHBG concentrations are reduced in women who subsequently develop pre-eclampsia.

Uterine artery Doppler and mid-trimester maternal plasma homocysteine in subsequent pre-eclampsia.

OBJECTIVE: To investigate whether mid-trimester maternal plasma homocysteine concentration is elevated in women who develop pre-eclampsia and in those women identified at high risk by abnormal uterine artery Doppler examination. METHODS: This was a multicenter study involving healthy women undergoing screening for pre-eclampsia by uterine artery Doppler velocimetry at 22-24 weeks' gestation. Abnormal uterine artery blood flow was defined as a mean pulsatility index (PI) above the 95th centile (1.6). Controls (mean PI < 1.6) were matched for gestational age and date of blood sample collection. Maternal plasma homocysteine concentration was measured retrospectively using a chemiluminescent immunoassay. RESULTS: In total, 683 women were recruited. Maternal plasma homocysteine concentration did not vary with gestation. Maternal plasma homocysteine concentration in women who subsequently developed pre-eclampsia (n = 80, 12%) was not significantly different from women with uncomplicated pregnancies (n = 536, 78%) (median 5.1, range 2.7-14.1 micromol/l vs. median 5.5, range 1.9-27.9 micromol/l, p = 0.44). There were no significant differences in the maternal plasma homocysteine concentration in women with abnormal uterine artery Doppler findings (n = 275) compared with controls (n = 408), (median 5.6, range 2.6-17.7 micromol/l vs. median 5.4, range 1.9-27.9 micromol/l, p = 0.13). CONCLUSION: Mid-trimester maternal plasma homocysteine concentration is not elevated in women who developed pre-eclampsia even in those at high risk defined by abnormal uterine artery Doppler velocimetry.

Second-trimester uterine artery Doppler screening in unselected populations: a review.

OBJECTIVE: Doppler ultrasound provides a non-invasive method for the study of the uteroplacental circulation. In normal pregnancy, impedance to flow in the uterine arteries decreases with gestation, which may be the consequence of trophoblastic invasion of the spiral arteries and their conversion into low-resistance vessels. Pre-eclampsia and fetal growth restriction are associated with failure of trophoblastic invasion of spiral arteries, and Doppler studies, in these conditions, have shown that impedance to flow in the uterine arteries is increased. A series of screening studies involving assessment of impedance to flow in the uterine arteries have examined the potential value of Doppler in identifying pregnancies at risk of the complications of impaired placentation. This review examines the findings of Doppler studies in unselected populations. METHODS: Searches of a computerized medical database were performed to identify relevant studies. Only those studies that provided sufficient data to allow calculation of the performance of the test were included in the analysis. Likelihood ratios were calculated for each study and are reported for pre-eclampsia, fetal growth restriction and perinatal death as well as for more severe forms of pre-eclampsia and fetal growth restriction. RESULTS: The literature search identified 19 relevant studies, four of which were excluded from the further analysis. The main characteristics and results of the 15 remaining studies provided discrepant results, which may be the consequence of differences in Doppler technique for sampling, the definition of abnormal flow velocity waveform, differences in the populations examined, the gestational age at which women were studied and different criteria for the diagnosis of pre-eclampsia and fetal growth restriction. Nevertheless, the studies provided evidence that increased impedance to flow in the uterine arteries is associated with increased risk for subsequent development of pre-eclampsia, fetal growth restriction and perinatal death. In addition, women with normal impedance to flow in the uterine arteries constituted a group that have a low risk of developing obstetric complications related to uteroplacental insufficiency. CONCLUSIONS: The review suggests that increased impedance to flow in the uterine arteries in pregnancies attending for routine antenatal care identifies about 40% of those who subsequently develop pre-eclampsia and about 20% of those who develop fetal growth restriction. Following a positive test, the likelihood of these complications is increased by about 6 and 3.5 times, respectively.

Maternal serum vitamin D levels at 11-13 weeks of gestation in preeclampsia.

This study is aimed to determine whether the maternal serum levels of vitamin D in the first trimester of pregnancy are altered in cases that develop preeclampsia (PE) and whether the levels are related to biochemical and biophysical markers of impaired placental perfusion and function. Maternal total serum vitamin D, pregnancy-associated plasma protein-A (PAPP-A), uterine artery pulsatility index (PI) and mean arterial pressure (MAP) were measured at 11-13 week gestation in 90 cases that developed PE, including 30 that required delivery before 34 weeks (early PE) and 1000 unaffected controls. The median values of vitamin D, PAPP-A, uterine artery PI and MAP expressed as a multiple of the unaffected median (MoM), in the patients developing early PE and late PE were compared with the controls. There was no significant difference in the median serum vitamin D MoM or raw values within the outcome groups (P=141 and P=0.231, respectively) whereas the median PAPP-A MoM, uterine PI MoM and MAP MoM were significantly different (P=0.031, P=0.001 and P<0.0001, respectively). Serum PAPP-A was decreased in both early PE and late PE (0.54 and 0.88 versus 1.03 MoM, P<0.0001 and P=0.010, respectively), MAP was increased in both early PE and late PE (1.09 and 1.06 versus 0.99 MoM, P<0.0001 and P<0.0001, respectively) and uterine artery PI was increased in early PE but not in late PE (1.32 and 1.12 versus 1.01 MoM, P<0.0001 and P=0.083, respectively). In pregnancies that subsequently develop PE maternal serum total vitamin D levels at 11-13 weeks are not altered.

Obesity in pregnancy.

Overweight and obesity are common findings in women of reproductive age in the UK; as 32% of 35- to 64-year-old women are overweight and 21% obese. Obesity causes major changes in many features of maternal intermediary metabolism. Insulin resistance appears to be central to these changes and may also be involved in increased energy accumulation by the fetus. Maternal obesity is associated with many risks to the pregnancy, with increased risk of miscarriage (three-fold) and operative delivery (20.7 versus 33.8% in the obese and 47.4% in the morbidly obese group). Other risks to the mother include an increased risk of pre-eclampsia (3.9 versus 13.5% in the obese group) and thromboembolism (0.05 versus 0.12% in the obese group). There are risks to the fetus with increased perinatal mortality (1.4 per 1000 versus 5.7 per 1000 in the obese group) and macrosomia (>90th centile; 9 versus 17.5% in the obese group). Maternal obesity is associated with an increased risk of obesity in the long term. Obese woman should try to lose weight before pregnancy but probably not during pregnancy. There is no real evidence base for the management of maternal obesity but some practical suggestions are made.

Relationship between second-trimester uterine artery Doppler and spontaneous early preterm delivery.

OBJECTIVE: To evaluate the relationship between impedance to flow in the uterine arteries at 22-24 weeks and subsequent spontaneous delivery before 33 weeks. METHODS: The pulsatility index (PI) in the uterine arteries was measured by transvaginal sonography at 22-24 weeks in 33,629 women with singleton pregnancies attending for routine antenatal care. The distribution of PI in the 237 patients with live births before 33 weeks, after spontaneous onset of labor, was compared to that in 31,633 patients with live births at or after 33 weeks. RESULTS: The median uterine artery mean PI was significantly higher in those women delivering before 33 weeks than in those delivering at or after 33 weeks. The mean PI was 1.57, which is the 95th centile, in 1525 (4.8%) of those delivering at or after 33 weeks and in 19 (8.0%) of the deliveries before 33 weeks. Multiple regression analysis demonstrated that the significant predictors of spontaneous delivery before 33 weeks were ethnic origin, smoking status, previous obstetric history and uterine artery mean PI. However, the prediction of spontaneous early preterm delivery by a combination of maternal characteristics and uterine artery Doppler was not significantly higher than maternal characteristics alone (area under the receiver-operating characteristics curve: 0.704, 95% CI 0.669-0.740 vs. 0.684, 95% CI 0.647-0.722; P = 0.062). CONCLUSIONS: Uterine artery PI in spontaneous deliveries before 33 weeks is higher than in those women delivering at or after 33 weeks. However, uterine artery Doppler does not provide a significant improvement in the prediction of spontaneous early delivery provided by maternal demographic characteristics and previous obstetric history.

Prediction of patient-specific risk of early preterm delivery using maternal history and sonographic measurement of cervical length: a population-based prospective study.

OBJECTIVE: To develop a model for calculating the patient-specific risk of spontaneous early preterm delivery by combining maternal factors and the transvaginal sonographic measurement of cervical length at 22 + 0 to 24 + 6 weeks, and to compare the detection rate of this method to that achieved from screening by cervical length or maternal characteristics alone. METHODS: This was a population-based prospective multicenter study involving 40,995 unselected women with singleton pregnancies attending for routine hospital antenatal care in London, UK. Complete follow-up was obtained from 39,284 (95.8%) cases. The main outcomes were detection rate, false-positive rate and accuracy of predicting spontaneous delivery before 32 weeks' gestation. RESULTS: Spontaneous delivery before 32 weeks occurred in 235 (0.6%) cases. The detection rate of screening for early preterm delivery, at a fixed false-positive rate of 10%, was 38% for maternal factors, 55% for cervical length and 69% for combined testing. There was good agreement between the model estimates and the observed probabilities of preterm delivery. CONCLUSIONS: This study provides a model that can give an accurate patient-specific risk of preterm delivery. The detection rate of screening by a combination of maternal factors and the measurement of cervical length was substantially higher than that of screening by each method alone.