Characterization of waste cell biomass derived glutamate decarboxylase for in vitro γ-aminobutyric acid production and value-addition.

Waste biomass of Lactobacillus brevis obtained from in vivo γ-aminobutyric acid (GABA) production was used for value-addition. This study aims to extract glutamate decarboxylase (GAD) and characterize it for in vitro GABA production. Extracted GAD showed an excellent activity for in vitro GABA production. 52 W ultrasonic output was best in crude GAD extraction which was purified by Q HP anion-exchange column followed by Superdex-200 colloid separation column. The molecular weight of the purified GAD was determined to be ~53 kDa, and the Km value for L-glutamic acid was calculated ~7.65 mM. Pyridoxal 5'-phosphate (PLP) acted as the best cofactor for GAD. Optimum temperature and PLP dosing were deferring for crude and purified enzyme forms which respectively exhibited at 45°C, 55°C, 200 µmol and 20 µmol whereas optimum pH was the same at 4.5. GAD finds applications in food industries hence its detailed characterization would be promising for commercial exploitations.

Low plasma levels of decoy receptor 3 (DcR3) in the third trimester of pregnancy with preeclampsia.

OBJECTIVE: The pathophysiology of preeclampsia, a major threat during pregnancy characterized by excessive inflammatory status, remains unclear. Decoy receptor 3 (DcR3), a soluble member of the tumor necrosis factor receptor (TNFR) superfamily, is capable of inducing anti-apoptosis via binding with TL1A and anti-inflammation by driving Th2 immune reactions. DcR3 may, therefore, play a role in immune modulation during pregnancy. The purpose of this study is to explore the role of DcR3 in normal and preeclamptic pregnancies. MATERIALS AND METHODS: Plasma samples from 104 normal pregnant women (26, 42, and 36 in the first, second, and third trimester, respectively) and 10 patients with preeclampsia in the third trimester were collected. Plasma DcR3 levels were determined by using commercial ELISA kits. ANOVA and linear regression analysis were performed to analyze the relationship between gestational age and DcR3 levels. After adjusting for gestational days, the levels of plasma DcR3 in preeclamptic and non-preeclamptic women in the third trimester were compared. RESULTS: The plasma levels of DcR3 gradually decreased as the gestational days increased during pregnancy (p < 0.05). In the third trimester, pregnant women with preeclampsia had significantly lower plasma DcR3 levels compared to non-preeclamptic women (p < 0.05). CONCLUSIONS: We found that plasma DcR3 levels gradually decreased as gestation progressed. The levels of plasma DcR3 in preeclamptic women were significantly lower than those of normal pregnant women, suggesting that a potential involvement of DcR3 in normal pregnancy and decreased levels of DcR3 may be related to preeclampsia.

Dynamic alterations of the levels of tumor necrosis factor-α, interleukin-6, and interleukin-1ß in rat primary motor cortex during transhemispheric functional reorganization after contralateral seventh cervical spinal nerve root transfer following brachial plexus avulsion injuries.

The transfer of a contralateral healthy seventh cervical spinal nerve root (cC7) to the recipient nerve in the injured side is considered a reliable and effective procedure for restoration of the physiological functions of an injured hand after brachial plexus root avulsion injury (BPAI). Growing evidence shows that the transhemispheric cortical reorganization is induced after cC7 nerve transfer surgery. However, little is known about the underlying molecular mechanism. Proinflammatory cytokines reportedly play an important role in the neural plasticity. We hypothesize that proinflammatory cytokines are involved in the transhemispheric functional reorganization after cC7 transfer. In the present study, we investigated the level of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1ß (IL-1ß) in the rat primary motor cortex after cC7 transfer following BPAI by enzyme-linked immunosorbent assay. The results showed that, in the sham group, no statistical significance was observed between the level of TNF-α, IL-6, and IL-1ß at each time point after the operation compared with that at day 0, respectively. However, in the unrepaired and repaired groups, the level of TNF-α, IL-6, and IL-1ß changed dynamically. The study is the first to provide evidence for the involvement of proinflammatory cytokines in transhemispheric functional reorganization after cC7 transfer following BPAI, which are useful for understanding the underlying mechanism.

Sphingosine-1-phosphate improves endothelialization with reduction of thrombosis in recellularized human umbilical vein graft by inhibiting syndecan-1 shedding in vitro.

Sphingosine-1-phosphate (S1P) has been known to promote endothelial cell (EC) proliferation and protect Syndecan-1 (SDC1) from shedding, thereby maintaining this antithrombotic signal. In the present study, we investigated the effect of S1P in the construction of a functional tissue-engineered blood vessel by using human endothelial cells and decellularized human umbilical vein (DHUV) scaffolds. Both human umbilical vein endothelial cells (HUVEC) and human cord blood derived endothelial progenitor cells (EPC) were seeded onto the scaffold with or without the S1P treatment. The efficacy of re-cellularization was determined by using the fluorescent marker CellTracker CMFDA and anti-CD31 immunostaining. The antithrombotic effect of S1P was examined by the anti-aggregation tests measuring platelet adherence and clotting time. Finally, we altered the expression of SDC1, a major glycocalyx protein on the endothelial cell surface, using MMP-7 digestion to explore its role using platelet adhesion tests in vitro. The result showed that S1P enhanced the attachment of HUVEC and EPC. Based on the anti-aggregation tests, S1P-treated HUVEC recellularized vessels when grafted showed reduced thrombus formation compared to controls. Our results also identified reduced SDC1 shedding from HUVEC responsible for inhibition of platelet adherence. However, no significant antithrombogenic effect of S1P was observed on EPC. In conclusion, S1P is an effective agent capable of decreasing thrombotic risk in engineered blood vessel grafts. STATEMENT OF SIGNIFICANCE: Sphingosine-1phosphate (S1P) is a low molecular-weight phospholipid mediator that regulates diverse biological activities of endothelial cell, including survival, proliferation, cell barrier integrity, and also influences the development of the vascular system. Based on these characters, we the first time to use it as an additive during the process of a small caliber blood vessel construction by decellularized human umbilical vein and endothelial cell/endothelial progenitor. We further explored the function and mechanism of S1P in promoting revascularization and protection against thrombosis in this tissue engineered vascular grafts. The results showed that S1P could not only accelerate the generation but also reduce thrombus formation of small caliber blood vessel.

Gene set-based integrative analysis of ovarian clear cell carcinoma.

OBJECTIVE: The pathogenesis of ovarian clear cell carcinoma is still poorly understood; therefore, we conducted a gene set-based analysis by integrating datasets downloaded from publicly available microarray gene expression databases to investigate the pathogenesis of clear cell carcinoma, which was based on the regularity of functions defined by gene ontology or canonical pathway databases. MATERIALS AND METHODS: The gene expression profiles of 80 clear cell carcinomas and 136 normal ovarian controls were downloaded from the National Center for Biotechnology Information Gene Expression Omnibus database. The gene expression profiles were converted to the gene set regularity (GSR) indexes computed using the modified differential rank conservation, an algorithm measuring the degree of gene expression ranking change in a gene set. Then the differences of GSR indexes between clear cell carcinomas and normal ovarian controls were analyzed. RESULTS: Machine learning can accurately recognize and classify the patterns of functional regularities containing the GSR indexes between the clear cell carcinomas and normal controls with an accuracy of 99.3%. The significant aberrations included oxidoreductase activity, binding, transport, channel activity, cell adhesion, immune response, chromosome assembly, and the deregulated signaling molecules, such as guanyl nucleotide exchange factors, phosphoinositide 3-kinase-activating kinase, receptor tyrosine kinase B, and protein tyrosine kinase. CONCLUSION: Our pioneering works using the functionome, which was converted from microarray gene expression profiles for integrative analysis, showed a clear distinction of functional changes between the clear cell carcinomas and normal ovarian controls. This approach might provide a comprehensive view of the deregulated functions of clear cell carcinomas for further investigation.

Gene Set-Based Integrative Analysis Revealing Two Distinct Functional Regulation Patterns in Four Common Subtypes of Epithelial Ovarian Cancer.

Clear cell (CCC), endometrioid (EC), mucinous (MC) and high-grade serous carcinoma (SC) are the four most common subtypes of epithelial ovarian carcinoma (EOC). The widely accepted dualistic model of ovarian carcinogenesis divided EOCs into type I and II categories based on the molecular features. However, this hypothesis has not been experimentally demonstrated. We carried out a gene set-based analysis by integrating the microarray gene expression profiles downloaded from the publicly available databases. These quantified biological functions of EOCs were defined by 1454 Gene Ontology (GO) term and 674 Reactome pathway gene sets. The pathogenesis of the four EOC subtypes was investigated by hierarchical clustering and exploratory factor analysis. The patterns of functional regulation among the four subtypes containing 1316 cases could be accurately classified by machine learning. The results revealed that the ERBB and PI3K-related pathways played important roles in the carcinogenesis of CCC, EC and MC; while deregulation of cell cycle was more predominant in SC. The study revealed that two different functional regulation patterns exist among the four EOC subtypes, which were compatible with the type I and II classifications proposed by the dualistic model of ovarian carcinogenesis.

Gene Set-Based Functionome Analysis of Pathogenesis in Epithelial Ovarian Serous Carcinoma and the Molecular Features in Different FIGO Stages.

Serous carcinoma (SC) is the most common subtype of epithelial ovarian carcinoma and is divided into four stages by the Federation of Gynecologists and Obstetrics (FIGO) staging system. Currently, the molecular functions and biological processes of SC at different FIGO stages have not been quantified. Here, we conducted a whole-genome integrative analysis to investigate the functions of SC at different stages. The function, as defined by the GO term or canonical pathway gene set, was quantified by measuring the changes in the gene expressional order between cancerous and normal control states. The quantified function, i.e., the gene set regularity (GSR) index, was utilized to investigate the pathogenesis and functional regulation of SC at different FIGO stages. We showed that the informativeness of the GSR indices was sufficient for accurate pattern recognition and classification for machine learning. The function regularity presented by the GSR indices showed stepwise deterioration during SC progression from FIGO stage I to stage IV. The pathogenesis of SC was centered on cell cycle deregulation and accompanied with multiple functional aberrations as well as their interactions.

Quantitative analysis of normal fetal medulla oblongata volume and flow by three-dimensional power Doppler ultrasound.

OBJECTIVE: Assessment of the fetal medulla oblongata volume (MOV) and blood flow might be important in the evaluation of fetal brain growth. We used three-dimensional power Doppler ultrasound (3DPDUS) to assess the fetal MOV and blood flow index in normal gestation. The relationships between these parameters were further analyzed. METHODS: We assessed the total volume and blood flow index of the fetal MO in normal pregnancies using a 3DPDUS (Voluson 730 Expert). The true sagittal plane over the fetal occipital area was measured by a 3D transabdominal probe to scan the fetal MO under the power Doppler mode. Then, we quantitatively assessed the total volume of the fetal MOV, mean gray area (MG), vascularization index (VI), and flow index (FI). RESULTS: A total of 106 fetuses, ranging from 19 weeks to 39 weeks of gestation, were involved in our study. The volume of the fetal MO was highly positively correlated with gestational age [correlation coefficient (r) = 0.686, p < 0.0001]. The MG was negatively correlated with gestational age [r = -0.544, p < 0.0001). VI and FI showed no significant correlation with gestational age (p = 0.123 and p = 0.219, respectively). CONCLUSION: 3DPDUS can be used to assess the fetal MOV and blood flow development quantitatively. Our study indicated that fetal MOV and blood flow correlated significantly with the advancement of gestational age. This information may serve as reference data for further studies of the fetal brain and blood flow under abnormal conditions.

Vaginal birth after cesarean section: 10 years of experience in a tertiary medical center in Taiwan.

OBJECTIVE: Because of the increased risk of uterine rupture and other morbidities, instances of trial of labor after cesarean (TOLAC) have decreased in number each year. Nevertheless, under careful assessment and advanced medical care, TOLAC is still a safe option for delivery. The objective of this study is to find the factors that impact the success rate for TOLAC and to compare the results with Taiwan national registry data. MATERIALS AND METHODS: A longitudinal cohort study that includes a total of 254 cases of women receiving TOLAC in a tertiary medical center over a period of 10 years. RESULTS: A total of 254 participants who underwent TOLAC, which accounts for 1.67% of total labor instances (254/15,166), were enrolled for analysis. The success rate of TOLAC was found to be 80.70% (205/254), including 146 (57.5%) normal deliveries, 45 (17.7%) vacuum-assisted deliveries, and 14 (5.5%) forceps-assisted deliveries. The conversion rate to cesarean section was 19.3%. There were no uterine rupture cases in our study, and there were only two suspected cases, which turned out to have no actual rupture. When analyzing the factors affecting the results of TOLAC, we found that a successfully spontaneously delivered baby had a lower birth weight than the failed TOLAC cases that were converted to cesarean delivery (mean, 2989 g vs. 3379 g; p < 0.001). Among the patients who were converted to cesarean section, the most common reason was dysfunctional labor (79.6%), followed by fetal distress (14.3%). CONCLUSION: Under intensive care and observation, TOLAC section may still be a feasible choice. Nevertheless, the body weight of the baby has been shown to be a factor that can influence the success rate.

Delivery of double singleton pregnancies in a woman with a double uterus, double cervix, and complete septate vagina.

Uterine anomalies involving a double uterus, double cervix, also known as didelphys uterus, and complete septate vagina are rarely seen and have an associated fertility problem. However, artificial reproductive technology with embryo transfers can help solve this fertility challenge. Conception in the uterus in just one side is commonly seen for embryos, which are always transferred through the usually used (dilated) vagina. We here present a patient with the above uterine anomaly who conceived with the aid of in vitro fertilization and embryo transfer to both uterine cavities under general anesthesia, which resulted in successful double singleton pregnancies with one fetus in each uterus. With intensive prenatal care, the pregnancy course for each fetus was rather uneventful. Although both fetuses were in cephalic presentation, cesarean section was performed at the 39(th) week of gestation with good outcomes in order to preclude anticipated difficulties if the baby had been delivered through the rarely dilated vagina. However, order of birth between the two fetuses was a crucial decision during the operation.

Noninvasive prenatal testing for fetal trisomy in a mixed risk factors pregnancy population.

OBJECTIVE: This study assesses the performance of noninvasive prenatal testing (NIPT) for fetal aneuploidies in a mixed risk factors pregnancy population. MATERIALS AND METHODS: Data review of 169 pregnant women undergoing prenatal aneuploidy screening in a single tertiary medical center was conducted. Indications included maternal anxiety, advanced maternal age, abnormal nuchal translucency, and high/moderate risk of first trimester Down syndrome screening. Multifetal pregnancies and patients receiving in vitro fertilization were also enrolled for analysis. RESULTS: A total of 169 patients were enrolled in this study during a time period from July 2012 to June 2014. For patients' ≥ 34 years, anxiety about amniocentesis was the most common reason for patients selecting NIPT for fetal aneuploidy screening, with 107 (88.4%) patients choosing NIPT for this reason. Among the total patient population, two patients showed a positive result from NIPT. One patient displayed 47, XXY, which was confirmed to be a false-positive result. The other patient displayed trisomy 18, which was confirmed by an amniotic cell culture. The sensitivity for NIPT is 100% with the specificity 99.4%. CONCLUSIONS: NIPT for fetal aneuploidy in a mixed risk factors pregnancy population showed high accuracy. NIPT applied to the low risk population might reassure the anxious family.

Pregnancy outcome of systemic lupus erythematosus in relation to lupus activity before and during pregnancy.

BACKGROUND: To analyze the pregnancy complications and pregnancy outcome relating to the disease activity of systemic lupus erythematosus before conception and during pregnancy. METHODS: Seventy-two pregnancies were collected in a single tertiary medical center within a 5-year period. Twelve pregnancies were terminated due to various causes in the first half of pregnancy. Analysis of pregnancy complications and pregnancy outcome relating to the lupus activity before conception and during pregnancy was made among the remaining 60 pregnancies with gestational length > 26 weeks. The assessment of lupus activity was based on the routine monitoring, including urine routine, white blood cell count, hemoglobin and platelet count, erythrocyte sediment rate, serum titers of C3, C4, and double-stranded DNA. Monitoring of renal function with daily urinary protein loss and clearance rate of creatinine was needed when worsened nephropathy was suspected. RESULTS: The etiologies of the terminated pregnancies were deteriorated nephropathy (6 cases), involvement of central nervous system (1 case), unwanted pregnancy due to drug exposure (3 cases), and two early intrauterine fetal deaths (both during the 23(rd) week of gestation). Pregnancy complications were related to the lupus activity before conception [odds ratio = 0.238, 95% confidence interval (CI) = 0.073, 0.778, p = 0.014] and during pregnancy (odds ratio = 0.153, 95% CI = 0.031, 0.754, p = 0.012). Meanwhile, pregnancy outcome significantly related to the lupus activity of the preconception period and during pregnancy. The gestational length was significantly longer in the pregnancies with remitted lupus activity either before conception (38.2 ± 1.6 weeks vs. 36.3 ± 3.4 weeks, p = 0.011 and 95% CI = -3.454, -0.478) or during pregnancy (38.2 ± 1.6 weeks vs. 35.2 ± 3.8 weeks, p = 0.005 and with 95% CI = -4.988, -1.005). Significant relationships were also found between newborn birth weights and lupus activity preconceptionally (2940 ± 389 g vs. 2448 ± 674 g, p = 0.002 and 95% CI = -792, -192) and after having conceived (2960 ± 383 g vs. 2136 ± 585 g, p < 0.001 and 95% CI = -1081, -568). Multivariate analysis showed that lupus remission during pregnancy was correlated with a significantly longer gestation, since pregnant women with active lupus had a three-fold greater risk of preterm deliveries (hazard ratio = 3.022, 95% CI = 1.261, 7.242) compared with pregnant women without active lupus. CONCLUSION: In order to reduce the incidence of pregnancy complication, especially preterm delivery, and to gain good pregnancy outcome, good preparation before conception and good control of the disease during pregnancy are mandatory.

Measurement of fetal maxillary and mandibular angles for first-trimester prenatal screening among Taiwanese women.

BACKGROUND: Early fetal structure evaluation is crucial. Fetal abnormalities might indicate chromosomal anomalies or abnormal fetal growth. The aim of this study was to establish the appropriate reference range of maxillary and mandibular angles among the Taiwanese population at 11(+0)-13(+6) weeks of gestation in normal singleton pregnancy as reference values for prenatal ultrasonographic examinations. METHODS: Fetal ultrasonographic data on maxillary angles and mandibular angles at a gestational age ranging from 11(+0) weeks to 13(+6) weeks were recorded in this study. Maternal background and pregnancy outcome were obtained from hospital records. RESULTS: A total of 87 patients were included in this study. Maxillary and mandibular angles were successfully recorded in 87 (100%) and 84 (96.6%) patients, respectively. The mean maternal age was 31 (range, 19-41) years, with a corresponding gestational age of 12(+4) (range, 11(+0)-13(+6)) weeks. The maxillary and mandibular angles were 79.9° ± 15.6° and 71.0° ± 12.8°, respectively. First-degree correlation was not found to exist between gestational age and maxillary and mandibular angles. CONCLUSION: Normative data for ultrasonographic measurements of maxillary and mandibular angles among the Taiwanese population are presented. Our results may serve as reference values in congenital anomaly screening during prenatal examination.

Assessment of placental volume and vascularization at 11-14 weeks of gestation in a Taiwanese population using three-dimensional power Doppler ultrasound.

BACKGROUND: The placental volume and vascular indices are crucial in helping doctors to evaluate early fetal growth and development. Inadequate placental volume or vascularity might indicate poor fetal growth or gestational complications. This study aimed to evaluate the placental volume and vascular indices during the period of 11-14 weeks of gestation in a Taiwanese population. METHODS: From June 2006 to September 2009, three-dimensional power Doppler ultrasound was performed in 222 normal pregnancies from 11-14 weeks of gestation. Power Doppler ultrasound was applied to the placenta and the placental volume was obtained by a rotational technique (VOCAL). The three-dimensional power histogram was used to assess the placental vascular indices, including the mean gray value, the vascularization index, the flow index, and the vascularization flow index. The placental vascular indices were then plotted against gestational age (GA) and placental volume. RESULTS: Our results showed that the linear regression equation for placental volume using gestational week as the independent variable was placental volume = 18.852 × GA - 180.89 (r = 0.481, p < 0.05). All the placental vascular indices showed a constant distribution throughout the period 11-14 weeks of gestation. A tendency for a reduction in the placental mean gray value with gestational week was observed, but without statistical significance. CONCLUSION: All the placental vascular indices estimated by three-dimensional power Doppler ultrasonography showed a constant distribution throughout gestation.

Umbilical artery Doppler velocimetry in normal pregnancies from 11(+0) to 13(+6) gestational weeks: a Taiwanese study.

OBJECTIVE: The aim of this study is to investigate the relationship between umbilical artery flow and gestational age (GA) at 11-13(+6) weeks in normal pregnancy in the Taiwanese population. MATERIALS AND METHODS: Two hundred and fifty-three normal singleton pregnancies with GA ranging from 11 to 13(+6) weeks were included in this study. The velocity of systolic, diastolic, and pulsatility index (PI) of the umbilical artery, and the mean velocity of the umbilical vein were recorded. RESULTS: One hundred and eighty-seven participants fulfilling the inclusion and exclusion criteria were analyzed. The mean maternal age was 31 (range 19-45) years, with a corresponding GA of 12(+4) (range 11-13(+6)) weeks. The absence of end-diastolic velocity of the umbilical artery was observed in most of our cases (90.1%). No significant change was found in the vascular indices with GA for the mean velocity of the umbilical vein (mean velocity = 0.923 × GA - 1.594, r = 0.1497, p = 0.115). The systolic velocity of the umbilical artery and PI, however, behaved differently from the other variables. There was a significant increase in systolic velocity with GA [systolic wave (S wave) velocity = 0.237 × GA + 2.267; r = 0.149, p = 0.041]. By contrast, the PI showed a significant decrease in relation to the GA (PI = -0.016 × GA + 4.068; r = 0.196, p = 0.007). CONCLUSION: The S-wave velocity of the umbilical artery increased with GA. By contrast, the PI of the umbilical artery showed a decreasing trend with GA. This may ensure optimal placental perfusion, which is necessary to accommodate the increased blood flow to the developing fetus. Furthermore, an absence of end-diastolic velocity in the first trimester and early second trimester was usually seen.