Enhancement of anaerobic biohydrogen/methane production from cellulose using heat-treated activated sludge.

Anaerobic digestion is an effective technology to convert cellulosic wastes to methane and hydrogen. Heat-treatment is a well known method to inhibit hydrogen-consuming bacteria in using anaerobic mixed cultures for seeding. This study aims to investigate the effects of heat-treatment temperature and time on activated sludge for fermentative hydrogen production from alpha-cellulose by response surface methodology. Hydrogen and methane production was evaluated based on the production rate and yield (the ability of converting cellulose into hydrogen and methane) with heat-treated sludge as the seed at various temperatures (60-97 degrees C) and times (20-60 min). Batch experiments were conducted at 55 degrees C and initial pH of 8.0. The results indicate that hydrogen and methane production yields peaked at 4.3 mmol H2/g cellulose and 11.6 mmol CH4/g cellulose using the seed activated sludge that was thermally treated at 60 degrees C for 40 min. These parameter values are higher than those of no-treatment seed (HY 3.6 mmol H2/g cellulose and MY 10.4 mmol CH4/g cellulose). The maximum hydrogen production rate of 26.0 mmol H2/L/d and methane production rate of 23.2 mmol CH4/L/d were obtained for the seed activated sludge that was thermally treated at 70 degrees C for 50 min and 60 degrees C for 40 min, respectively.

Bax, Bak and mitochondrial oxidants are involved in hypoxia-reoxygenation-induced apoptosis in human placenta.

Although apoptosis is prominent in placental cells in pregnancy complications such as preeclampsia, the cause is unknown. We surmised that hypoxia-reoxygenation (HR) is the mechanism and hypothesized that mitochondrial oxidants and Bcl-2 proteins cause HR-induced placental apoptosis. Our goal was studying expression of five Bcl-2 proteins--Bcl-2, Bcl-xL, Bax, Bak, Bad--and testing effects of diazoxide and cyclosporine A on oxidative stress and apoptosis in villous tissues subjected to HR. Term human placentas were obtained from normal pregnancies following elective caesarean deliveries. Villous tissues were subjected to "repetitive HR" (one hour at 2% O(2) then one hour at 8% O(2), alternatively, for a total of 6h) or "prolonged HR" (3h at 2% O(2) then 3h of 8% O(2)). Samples maintained at 2% and 8% O(2) served as hypoxic and normoxic controls, respectively. Prolonged HR caused the most severe villous apoptotic changes, increased the expression of Bax and Bak mRNA and protein and reduced the expression of Bcl-2 mRNA. Pre-administration of diazoxide and cyclosporine A reduced TUNEL-positive nuclei and levels of nitrotyrosine and 4-hydroxy-2-nonenol after prolonged HR. Thus, duration of hypoxia and reoxygenation is important in determining severity of HR-induced apoptosis in placenta. These apoptotic changes are closely associated with Bax and Bak effects and oxidative stress in mitochondria.

Tumour necrosis factor-alpha converting enzyme in human gestational tissues from pregnancies complicated by chorioamnionitis.

Chorioamnionitis increases the risk of preterm labour and is associated with adverse neonatal outcomes including cerebral palsy. Tumour necrosis factor-alpha (TNF-alpha) derived from the gestational tissues (placenta, fetal membranes and maternal decidua) is thought to play a pivotal role in the induction of cytokine response in chorioamnionitis. Tumour necrosis factor-alpha converting enzyme (TACE) is essential for the release of TNF-alpha. Our aim was to determine whether the expression of TACE is increased in human gestational tissues from pregnancies complicated by chorioamnionitis, and whether lipopolysaccharide (LPS) causes increased expression of TACE in the human gestational tissues in vitro. The immunostaining of TACE was generally more intense, in particular in the syncytiotrophoblast and stromal cells, in villous samples from pregnancies complicated by chorioamnionitis than those from normal pregnancies. Increased immunoreactivity of TACE was also noted in the amnion and choriodecidua. In parallel, there was an increased infiltration of monocytes/macrophages within the villous stroma and choriodecidua. As a complement to our in vivo findings, LPS significantly increased the levels of mRNA and protein of TACE in a dose-dependent response in villous and fetal membrane explant cultures. Together, our results imply a potential role of TACE in the pathogenesis of chorioamnionitis.

Detection of aflatoxin B1-DNA adducts in human placenta and cord blood.

Human placenta and cord blood are readily available specimens that respond to maternal environmental insult and are being used to investigate metabolism, bioactivation, and transplacental transfer of procarcinogens. Enzyme-linked immunosorbent assay was used to quantitate 120 placentas and 56 cord bloods from term, uncomplicated pregnancies at Taipei Chang Gung Memorial Hospital, Taiwan, for the presence of the imidazole ring-opened form of aflatoxin B1-DNA (AFB1-DNA) adducts. Of the 120 samples of placentas, 69 (57.5%) contained AFB1-DNA adducts in levels from 0.6 to 6.3 mumol/mol DNA. Of the 56 samples of cord bloods, 5 (8.9%) contained AFB1-DNA adducts in levels from 1.4 to 2.7 mumol/mol DNA. A higher positive rate was found in samples collected in the summer than in the winter. These results indicate that a significant number of individuals in an area of high liver cancer risk have been exposed to AFB1, and it is possible to transfer AFB1 and its metabolites to the progeny through the transplacental unit. Thus, monitoring adduct levels in human specimens may provide information not only on carcinogen exposure but also on the relationship among infection with hepatitis B/C virus, dietary exposure to AFB1, and liver cancer.

Outcome of congenital cystic adenomatoid malformation of the lung after antenatal diagnosis.

OBJECTIVE: We evaluated the outcome of fetuses diagnosed with having congenital cystic adenomatoid malformation (CCAM) on ultrasonographic examination and managed conservatively. METHODS: A retrospective study of 19 cases of CCAM diagnosed antenatally in our hospital was conducted between 1990 and 2001. Complete clinical information was available for all patients, with a mean follow-up of 62 months. RESULTS: The median gestational age at which CCAM was diagnosed was 23 weeks and there were eight live births. With conservative postnatal management, seven neonates had no major complications and one developed bronchopneumonia. CONCLUSION: Taken together, the findings of the present study and a review of the literature strongly support the conservative management of selected neonates with CCAM.

Differential expression of interleukin-1 beta and interleukin-6 in human fetal serum and meconium-stained amniotic fluid.

The study was designed to investigate the expression of the inflammatory cytokines interleukin-1 beta and interleukin-6 in meconium-stained amniotic fluid and in fetal cord serum. Amniotic fluid and fetal cord serum specimens were collected from 10 and 9 women with meconium-stained and clear amniotic fluid, respectively, during Caesarean operation at labor The mean concentrations of interleukin-1 beta found in clear and meconium-stained amniotic fluid were 10.0 and 54.5 pg/ml, respectively, and the difference was not statistically significant. On the other hand, the concentrations of interleukin-6 in meconium-stained amniotic fluid (774 pg/ml) was significantly higher than that found in clear amniotic fluid (149 pg/ml) (P = 0.0036). The differences of levels of both interleukin-1 beta and interleukin-6 in fetal cord serum specimens were not significant between neonates born to mothers with either clear or meconium-stained amniotic fluid (P = 0.8702 and 0.2987, respectively). The results of this study suggest that the production of at least one of the inflammatory cytokines, interleukin-6, is associated with the meconium found in amniotic fluid.

Hepatitis C virus in peripheral blood mononuclear cells.

RNA extracted from plasma and peripheral blood mononuclear cells of patients with chronic hepatitis C were used as the template for reverse transcription followed by double in vitro enzymatic amplification with nested primers. Hepatitis C virus was detected in 14 of 15 (93.3%) plasma specimens and in 8 of 15 (53.3%) peripheral blood mononuclear cell specimens obtained from patients with chronic hepatitis C and abnormal liver functions. The results suggest that hepatitis C virus could be found frequently in peripheral blood mononuclear cells of patients with chronic hepatitis C. Whether the presence of hepatitis C virus in peripheral blood mononuclear cells plays any role in the pathogenesis of diseases associated with hepatitis C virus infection remains to be determined.

Identification of human papillomavirus DNA sequences in peripheral blood mononuclear cells.

Polymerase chain reaction (PCR) was used to amplify and identify the presence of the DNA of human papillomavirus (HPV) types 6, 11, 16, and 18 in peripheral blood mononuclear cells (PBMCs) of women with and without urogenital HPV infections. HPV DNA of various types was found in PBMCs of 13 of 25 (52.0%) patients with urogenital HPV infections and in none of the 19 control subjects who are free of urogenital HPV infections. The presence of HPV DNA in PBMCs may impair the immunologic functions of the lymphocytes and play a role in the epidemiology of HPV infections and the pathogenesis of HPV-induced diseases.

Presence of cells of fetal origin in maternal circulation of pregnant women.

Fetal cells can be identified by using the polymerase chain reaction to test for the presence of human Y-chromosome-specific ZFY and SRY gene DNA sequences in maternal peripheral blood of women who bear a male fetus. Thirty-one pregnant women were studied in the first trimester to determine when fetal cells become detectable in the maternal circulation. Among the 19 women whose peripheral blood samples were positive for Y-chromosome-specific DNA sequences, the presence of fetal cells was quite case-variable from the 6th to 12th gestational weeks. Twenty-eight women who had given birth to their first male babies were studied postpartum to determine when fetal cells disappear from the maternal circulation. Fetal cells can still be detected in maternal blood 10 months postpartum in some cases. These results suggest that identification of fetal cells in the maternal circulation is possible. Nevertheless, interpretation of fetal cells in maternal circulation should be handled very carefully with respect to when these fetal cells first became detectable and potential interference from previous pregnancies.

Down syndrome screening in an Asian population using alpha-fetoprotein and free beta-hCG: a report of the Taiwan Down Syndrome Screening Group.

OBJECTIVE: To evaluate whether the strategy of maternal serum screening for Down syndrome, using alpha-fetoprotein (AFP) and free beta-hCG in combination with maternal age, a technique developed in western countries, is applicable to an Asian population. METHODS: Alpha-fetoprotein and beta-hCG were measured in serum samples from 23 Down syndrome pregnancies and 1748 unaffected singleton Taiwanese (ethnically Chinese) pregnancies at 14-22 weeks' gestation. Gestational age-specific medians and a maternal weight correction formula were established for our own population. Likelihood ratio for Down syndrome pregnancies in relation to multiples of the median (MoM) levels of these analytes were derived from the overlapping gaussian frequency distribution curves for Down syndrome and unaffected pregnancies. RESULTS: The serum AFP and free beta-hCG median MoM values of Down syndrome pregnancies were significantly abnormal in Asian subjects (0.77 and 2.91, respectively), and similar to those of affected pregnancies in white women. The median value of free beta-hCG:AFP MoM ratio (2.97) in Down syndrome pregnancies was significantly higher than that of unaffected pregnancies (1.09). The mean maternal weight during the second trimester in pregnant Asian women (55.2 kg) was markedly lighter than that of white women. At a 5.8% false-positive rate, free beta-hCG identified 47.8% of Down syndrome pregnancies (likelihood ratio 8.2), AFP detected only 13% of the cases (likelihood ratio 2.2), and free beta-hCG:AFP MoM ratio detected 43.5% of the cases (likelihood ratio 7.4). By using a multivariate risk algorithm involving the combination of AFP, free beta-hCG, and maternal age, 56.5% of Down syndrome cases could be detected with a 5.3% false-positive rate (likelihood ratio 10.7). CONCLUSION: Maternal serum screening strategy using AFP and free beta-hCG in combination with maternal age is feasible in the detection of fetal Down syndrome among Asian women.

Prediction of adverse perinatal outcome by maternal serum screening for Down syndrome in an Asian population.

OBJECTIVE: To investigate the association between adverse perinatal outcomes and abnormal elevations of serum marker levels (alpha-fetoprotein [AFP] and free beta-hCG) or a false-positive screen for Down syndrome. METHODS: Pregnancy outcome information was available for 5885 Taiwanese women under 35 years of age who had second-trimester maternal serum screening for Down syndrome, using AFP and free beta-hCG, and delivered a chromosomally normal fetus. Those with AFP at least 2.0 multiples of the median (MoM), free beta-hCG at least 2.5 MoM, or a false-positive screen (risk ratio at least 1:270) were identified, and the risk for adverse perinatal outcome was assessed. RESULTS: A serum AFP level at least 2.0 MoM (n = 176, 3.0%) was significantly associated with the occurrence of preterm delivery, low Apgar scores, small-for-gestational-age infants, low birth weight or very low birth weight, fetal death, premature rupture of membranes, oligohydramnios, and a higher incidence of perinatal mortality. A serum free beta-hCG level at least 2.5 MoM (n = 416, 7.1%) was significantly associated with low birth weight, an abnormally adherent placenta, and the occurrence of meconium-stained amniotic fluid. A higher incidence of fetal structural anomalies other than neural tube or abdominal wall defects, large-for-gestational-age infants, and postpartum hemorrhage was observed for a calculated risk of at least 1:270 (n = 311, 5.3%) independent of the other biochemical markers. CONCLUSION: Asian women with unexplained elevations of serum AFP or free beta-hCG, or a false-positive screen for Down syndrome are at increased risk for various adverse perinatal outcomes. Careful fetal ultrasound examination and thoughtful strategy for perinatal management are warranted for these patients.

Urine free beta-hCG and total estriol for Down syndrome screening during the second trimester in an Asian population.

OBJECTIVE: To evaluate second-trimester free beta-hCG and total estriol (E3) in the maternal urine as markers for Down syndrome screening in an Asian population. METHODS: Free beta-hCG and total E3 were measured in the urine samples of 28 Taiwanese Down syndrome pregnancies and 268 unaffected singleton pregnancies at 14-25 weeks. Results were normalized to urine creatinine concentrations and converted to multiples of the median (MoM) levels. Gestational ages were estimated by ultrasound measurements. RESULTS: Median values of free beta-hCG, total E3, free beta-hCG to total E3 ratio, and the free beta-hCG to total E3 MoM ratio in Down syndrome pregnancies were 4.75 MoM, 0.66 MoM, 8.99 MoM, and 9.51, respectively. At a 5% false-positive rate, the observed detection rates were 36% (ten of 28) with total E3, 71% (20 of 28) with free beta-hCG, 68% (19 of 28) with free beta-hCG/total E3, and 71% (20 of 28) with free beta-hCG/total E3 MoM. When combined with maternal age, the expected detection rates were 65% with total E3, 71% with free beta-hCG, 76% with free beta-hCG/total E3, 80% with free beta-hCG/total E3 MoM, and 89% when combining free beta-hCG, total E3, and maternal age. CONCLUSION: Urine free beta-hCG and total E3 are useful markers for Down syndrome screening during the second trimester in Taiwanese women.

Risk factors for placenta accreta.

OBJECTIVE: To identify risk factors associated with placenta accreta in a large cohort study. METHODS: Data for this study came from the Taiwan Down Syndrome Screening Group, an ongoing project on feasibility of serum screening in an Asian population. Women who had serum screening for Down syndrome at 14-22 weeks' gestation using alpha-fetoprotein (AFP) and free beta-hCG between January 1994 and June 1997, and delivered in the same institution, were included (n = 10,672). Those who had multiple gestations (n = 200), overt diabetes (n = 11), or fetal malformations (n = 101) were excluded. If a woman was involved more than once, one randomly selected pregnancy was included in the analysis (n = 9349). Twenty-eight pregnancies were complicated by placenta accreta, diagnosed by clinical presentation (n = 26) or histologic confirmation (n = 2). Multiple logistic regression with adjustment for potentially confounding variables was used to identify independent risk factors for placenta accreta. RESULTS: Women who had placenta previa (odds ratio [OR] 54.2; 95% confidence interval [CI] 17.8, 165.5) and second-trimester serum levels of AFP and free beta-hCG greater than 2.5 multiples of the median (OR 8.3; 95% CI 1.8, 39.3 and OR 3.9; 95% CI 1.5, 9.9, respectively), and were 35 years and older (OR 3.2; 95% CI 1.1, 9.4) were at increased risk of having placenta accreta. CONCLUSION: Risk factors for placenta accreta include placenta previa, abnormally elevated second-trimester AFP and free beta-hCG levels, and advanced maternal age.

Use of the antral follicle count to predict the outcome of assisted reproductive technologies.

OBJECTIVE: To evaluate the predictive value of the antral follicle count in patients undergoing assisted reproductive technologies (ARTs). DESIGN: Prospective study. SETTING: Tertiary care institutional hospital. PATIENT(S): Consecutively seen patients undergoing ARTs such as IVF-ET, gamete intrafallopian transfer, and tubal embryo transfer (TET). INTERVENTION(S): The ovarian antral follicle number was determined by transvaginal ultrasonography on the first or second menstrual day, before the administration of gonadotropins, in patients undergoing ARTs. MAIN OUTCOME MEASURE(S): Ovulation induction was accomplished with the use of GnRH agonist down-regulation combined with FSH and menotropin stimulation. Gamete intrafallopian transfer or TET was performed in patients with patent fallopian tubes, and IVF-ET was undertaken in the remaining patients. Analysis of variance and Mantel-Haenszel monotonic test for trends were used for data analysis. RESULT(S): A total of 149 treatment cycles for 130 couples were performed during the study period. The procedures performed included 89 ETs, 26 gamete intrafallopian transfers, 13 TET cycles, and 21 incomplete cycles (9 poor responders, 6 failed retrievals, and 6 nonfertilization cycles). All treatment cycles were divided into three groups according to the number of antral follicles (i.e., < or = 3, 4-10, and > or = 11) to evaluate the influence of various factors. The antral follicle count correlated significantly with patient age, day 3 serum FSH level, use of gonadotropins, serum estradiol concentration, number of oocytes retrieved, and, later, number of oocytes or embryos transferred. The group of patients who had a lower antral follicle count also had a significantly higher rate of cycle cancellation compared with the other two groups (68.8% vs. 5.3% and 0, respectively). No pregnancies occurred in the low antral follicle count group, whereas there was a trend toward an increasing number of pregnancies per attempt as the number of antral follicles increased (0, 23.7%, and 36.8%, respectively). CONCLUSION(S): It is easy to determine the number of antral follicles with a diameter of 2-5 mm on the first or second day of menstruation, or just before the administration of exogenous gonadotropins. We were able to predict the ovarian response and pregnancy results of patients undergoing ARTs with the use of this simple procedure.

Hepatitis B virus DNA in cervicovaginal cells.

Hepatitis B virus DNA sequences were detected in seven (12.1%) of 58 cervicovaginal cell specimens that were obtained from pregnant women by polymerase DNA amplification assay. The presence of hepatitis B virus DNA in these cells raises the possibility that infected cervicovaginal cells may be a source through which hepatitis B virus can be transmitted from infected mothers to their newborns and between heterosexual partners.

Risk factors for pre-eclampsia in an Asian population.

OBJECTIVE: To identify the risk factors for pre-eclampsia in an Asian population. METHOD: We conducted a retrospective cohort study involving 29375 Taiwanese women who delivered between July 1990 and September 1998, excluding pregnancies complicated by chronic hypertension or fetal malformations. RESULT: Four hundred and fifteen women had pre-eclampsia (1.4%). Women who had a history of pre-eclampsia (OR 6.3, 95% CI 4.4, 9.2), multiple gestation (OR 3.6, 95% CI 2.4, 5.5), a prepregnancy BMI > 24.2 kg/m(2) (OR 2.4, 95% CI 1. 8, 3.1), were > 34 years of age (OR 1.8, 95% CI 1.4, 2.4), nulliparous (OR 1.3, 95% CI 1.2, 1.5), had urinary tract infection (OR 4.8, 95% CI 1.5, 15.8), or worked during pregnancy (OR 1.9, 95% CI 1.4, 2.4) were at increased risk of pre-eclampsia. CONCLUSION: Some of the risk factors for pre-eclampsia among Asian women are the same as those of other ethnic groups, whereas some of the risk factors are different.

Maternal renal artery Doppler flow-velocity waveform in preeclampsia. A preliminary report.

By use of duplex Doppler ultrasonography, we preliminarily studied 30 normal pregnancies (control group) and 30 patients with preeclampsia (study group) before treatment. The peak systolic flow height to end diastolic flow height (S/D) ratio of the maternal right and left renal arteries of both groups were compared and analyzed. The results showed that there were no significant differences between the right and left renal arteries in either group, but unexpectedly the S/D ratio of the renal artery on both sides in preeclamptic patients was significantly lower than in normal pregnant women (mean +/- SD, 2.07 +/- .12 vs. 2.41 +/- .12, respectively, P < .001). The lower S/D ratio of the renal artery may reflect a decrease in renovascular resistance and an increase in renal perfusion. These findings were the reverse of what might be expected from an elevated vascular resistance and hypoperfusion model.

Endocardial cushion defect in a fetus: with intrauterine cytomegalovirus infection and growth retardation: report of an autopsy case.

We describe one case of endocardial cushion defect during the second trimester when a consultative fetal echocardiography was undertaken for fetal intrauterine growth retardation with oligohydramnios. Positive cytomegalovirus IgM in cord blood and cytomegalovirus DNA particles in the amniotic fluid were found incidentally. The relationship between possible intrauterine cytomegalovirus infection and the endocardial cushion defect in this case is not clear.

Birth weight by gestational age in twin pregnancies: analysis of 661 pairs.

The mortality of twin infants is four to five times higher than that of singletons, and one-half to two-thirds of all twins weigh < 2,500 g at birth. The appropriate interpretation of fetal growth throughout pregnancy is dependent upon the availability of adequate standards. We reviewed 661 pairs of live twin infants born at Chang Gung Memorial Hospital from 1979 to 1990. The frequency of twin births was 1.17% (1:86), and the ratio of males to females was 1.03. The frequency of preterm births (< 37 weeks) was 36.9%, the frequency of low birth weight (< 2,500 g) was 47.9% and very low birth weight (< 1,500 g) was 6.7%. A fetus grows most rapidly from the 32nd to the 35th week of gestation (200 g per week). The growth was 145 g per week from the 28th to the 32nd week and from the 35th to the 38th week of gestation. After the 38th week, the mean birth weight increased by only 35 g per week. Compared with a singleton birth, the mean birth weight of twins was about 100 g lighter during the 28th to the 32nd week, then the difference increased gradually to about 500 g at term.

In vitro perfusion study of the human placenta at term: preliminary results.

Dual perfusion of the human placental lobule in vitro is a useful method for studying the transfer of molecules across the placenta, including the transfer of endogenous substances and xenobiotics. To establish the first model of in vitro placental dynamic study in our country, we used a dual recirculating perfusion system modified from that described by Miller et al. A placental lobule without tears or gross lesions was chosen. Both the fetal and maternal sides of the placenta were perfused with Medium 199 in addition to heparin, glucose, dextran and antibiotics. Perfusate samples were obtained periodically and analyzed for blood gas, glucose, lactate, human placental lactogen (hPL) and human chorionic gonadotropin (hCG). The stability of this human placental lobule preparation during 10 hours of perfusion was reflected by the stability of the arterial pressure and by the constant volume in the fetal compartment. A constant rate of oxygen was delivered by the maternal circulation system, and a steady rate of oxygen was gained by the fetal circulation system. Neither oxygen nor glucose consumption by the tissue was significantly reduced during the period of perfusion. The releasing rates of hCG and hPL did not change significantly during perfusion. The development of this dual perfusion system for the human placenta can provide for the study of placental hemodynamics and transplacental transport in perinatal medicine in our country.