Comparisons of urine protein-to-creatinine ratios and their dynamic change patterns during labor at term between normal pregnant women and women with pregnancy induced hypertension.

Introduction: To evaluate patterns of change in the urine protein-to-creatinine ratios (uPCRs) during labor at term between normal and women with pregnancy-induced hypertension (PIH). Methods: This is an observational study in tertiary referral hospital, recruiting 269 women at term delivery in Taiwan from April 19, 2019 to April 18, 2021. uPCRs in four phases (latent, active, recovery and early postpartum) and related clinical data at delivery were collected. Multivariate analyses with a linear regression model were performed to analyze continuous variables after adjusting for clinical data between two groups. Results: Based on exclusion criteria, 68 normal and 24 pregnant women with PIH were included. There were no differences in the uPCR or the proportion cases of uPCRs ≥ 300 mg/g between normal and PIH group in the four phases. There was a statistically significant tendency for the proportion of uPCRs ≥ 300 mg/g to increase from the latent to the early postpartum phase in both groups. The proportion of uPCRs ≥ 300 mg/g significantly increased from the active to the recovery phase and then declined from the recovery to the early postpartum phase in the normal group. Thus no differences in uPCRs cases change between any two phases in women with PIH, except the duration above stated. Conclusion: This is the first study to demonstrate that uPCRs data are not different between normal pregnant and PIH groups during the course of labor, but it did show different dynamic change patterns throughout the labor phases.

Quality Assessment of Meta-Analyses on Cervical Cancer.

Meta-analysis (MA) quantitatively summarizes the findings of independent studies and is considered the highest quality of evidence for evidence-based medicine. However, issues in reporting and methodological rigor of MA hamper reproducibility and create the potential for bias. By applying PRISMA reporting guideline and AMSTAR2 execution guidelines on 40 cervical cancer MA samples covering topics such as interventions and risk factors, we determined the extent to which MA execution adhered to best practice guidelines. The results show that the elements with least adherence include "review methods established before MA" and "principal summary measures defined" (each 32.5% per PRISMA) and "characteristics of included studies" (31.3% per AMSTAR2) which undermine reproducibility and increase the risk of bias. This initial work presents common pitfalls in MA and is intended to improve awareness of these issues for clinicians who are interested in conducting MA and to pave the way toward quality improvement via informatics approaches.

T-complex protein 1 subunit zeta-2 (CCT6B) deficiency induces murine teratospermia.

BACKGROUND: The CCT complex is an important mediator of microtubule assembly and intracellular protein folding. Owing to its high expression in spermatids, CCT knockdown can disrupt spermatogenesis. In the present report, we therefore evaluated the in vivo functionality of the testis-specific CCT complex component CCT6B using a murine knockout model system. METHODS: A CRISPR/Cas9 approach was used to generate Cct6b-/- mice, after which candidate gene expression in these animals was evaluated via qPCR and Western blotting. Testicular and epididymal phenotypes were assessed through histological and immunofluorescent staining assays, while a computer-assisted sperm analyzer was employed to assess semen quality. RESULTS: Cct6b-/- mice were successfully generated, and exhibited no differences in development, fertility, appearance, testis weight, or sperm counts relative to control littermates. In addition, no differences in spermatogenesis were detected when comparingCct6b+/+ and Cct6b-/- testes. However, when progressive motility was analyzed, the ratio of normal sperm was significantly decreased in Cct6b-/- male mice, with nuclear base bending being the primary detected abnormality. In addition, slight decreases in Cct4 and Cct7 expression were detected. CONCLUSION: These data indicated that CCT6B is an important regulator of murine spermatogenesis, with the loss of this protein resulting in CCT complex dysfunction, providing a foundation for further studies.

A joint method for the screening of pharmacological chaperones for phenylalanine hydroxylase.

Phenylalanine hydroxylase (PAH) deficiency (PAHD) is an autosomal recessive disorder that causes severe injury to the nervous system, the treatment of which mainly depends on dietary therapy. The limited treatment options for PAHD are an incentive to develop new methods to identify more efficient therapeutic drugs, such as agonists which could improve PAH activity. In this study, we aimed to establish a rapid and convenient method for the screening and verification of PAH agonists. We compared fluorospectrophotometry and tandem mass spectrometry for detection of enzymatic formation of tyrosine, finding that the latter was a more sensitive method. We optimized immunoprecipitation purification conditions and measurement conditions of PAH activity. The optimal ratio between PAH protein and magnetic beads was 500 µg protein per 20 µL beads, and the optimized conditions for the detection of PAH enzymatic activity included the presence of 75 µM coenzyme ((6R)-l-erythro-5,6,7,8-tetrahydrobiopterin) and 30 min reaction time. Based on virtual screening, we screened ten candidate agonists from the FDA drug library. Three of these (nefopam, fluocinonide, and risperidone) were found to activate the enzyme in a dose-dependent manner (0.1-10 µM) by the joint method. We tested the efficacy of the three agonists on three PAH mutations (p.I65T, p.H107R, and p.D101N) that influence enzyme activity, and found that risperidone could specifically activate D101N-mutated enzyme. In conclusion, we established a joint method that is highly reliable, cost-effective, labor-saving, and time-saving. And we also found a specific agonist for D101N-mutated PAH by this joint method which may assist the development of clinical treatment for PAHD patients with different enzyme deficiencies.

Examining Reproducibility of Literature Search in Meta-Analysis.

Meta-analysis, a systematic retrieval from literature databases is an essential and prevailing method for combining data from multiple studies. Unfortunately, few studies have examined its rigor, which affects its reproducibility of results. We identified 22 meta-analyses on cervical cancer in PubMed for examining the parameters defined by PRISMA, relating to the rigor of literature retrieval. We found that 16 literature databases were used, and EMBASE was a leading resource, accounting for the highest frequency (81.82%). About half (45.45%) of the meta-analyses presented a complete, reproducible search strategy for at least one database. The ratio of included to retrieved articles after redundancy removal was only 6.58%, indicating low precision due to unclear or unreported processes. Our work serves as an initial step to examine the planning and execution of meta-analysis. Future efforts need to enhance reliability on literature retrieval in meta-analysis and compliance to PRISMA.

Prenatal diagnosis of radial ray defects by ultrasound: A report of 6 cases.

OBJECTIVE: All of the medical records of fetuses with the sonographic finding of radial ray defects (RRDs) between 2008 and 2015 were retrieved. The associated sonographic findings, cytogenetic results, and necropsy findings were correlated. CASE REPORT: There were 6 cases of RRD. Three cases were bilateral and the other 3 cases were unilateral. The gestational ages at diagnosis were between 12 and 24 weeks gestation. All women carrying fetuses with RRDs opted to terminate the pregnancy. There were 2 cases of trisomy 18, one case of thrombocytopenia-absent radius syndrome, and 2 cases of isolated RRD. Both cases of trisomy 18 had other sonographic abnormalities. CONCLUSION: RRD should be considered if a short radius and abnormal angulation of the wrist or thumb is noted. The use of 3-D ultrasound facilitates the diagnosis of RRD, even at early gestation, by providing a better surface appearance, panoramic views, and spatial orientation.

Comparison of the Effects of Daptomycin on Bacterial and Model Membranes.

Daptomycin is a phosphatidylglycerol specific, calcium-dependent membrane-active antibiotic that has been approved for the treatment of Gram-positive infections. A recent Bacillus subtilis study found that daptomycin clustered into fluid lipid domains of bacterial membranes and the membrane binding was correlated with dislocation of peripheral membrane proteins and depolarization of membrane potential. In particular, the study disproved the existence of daptomycin ion channels. Our purpose here is to study how daptomycin interacts with lipid bilayers to understand the observed phenomena on bacterial membranes. We performed new types of experiments using aspirated giant vesicles with an ion leakage indicator, making comparisons between daptomycin and ionomycin, performing vesicle-vesicle transfers, and measuring daptomycin binding to fluid phase versus gel phase bilayers and bilayers including cholesterol. Our findings are entirely consistent with the observations for bacterial membranes. In addition, daptomycin is found to cause ion leakage through the membrane only if its concentration in the membrane is over a certain threshold. The ion leakage caused by daptomycin is transient. It occurs only when daptomycin binds the membrane for the first time; afterward, they cease to induce ion leakage. The ion leakage effect of daptomycin cannot be transferred from one membrane to another. The level of membrane binding of daptomycin is reduced in the gel phase versus the fluid phase. Cholesterol also weakens the membrane binding of daptomycin. The combination of membrane concentration threshold and differential binding is significant. This could be a reason why daptomycin discriminates between eukaryotic and prokaryotic cell membranes.

Action of Antimicrobial Peptides on Bacterial and Lipid Membranes: A Direct Comparison.

The bacterial membrane represents an attractive target for the design of new antibiotics to combat widespread bacterial resistance. Understanding how antimicrobial peptides (AMPs) and other membrane-active agents attack membranes could facilitate the design of new, effective antimicrobials. Despite intense study of AMPs on model membranes, we do not know how well the mechanism of attack translates to real biological membranes. To that end, we have characterized the attack of AMPs on Escherichia coli cytoplasmic membranes and directly compared this action to model membranes. AMPs induce membrane permeability in E. coli spheroplasts or giant unilamellar vesicles (GUVs) under well-defined concentrations of AMPs and fluorescent molecules. The action of AMPs on spheroplasts is unique in producing an intracellular fluorescence intensity time curve that increases in a sigmoidal fashion to a steady state. This regular pattern is reproducible by melittin, LL37, and alamethicin but not by CCCP or daptomycin, agents known to cause ion leakage. Remarkably, a similar pattern was also reproduced in GUVs. Indeed the steady-state membrane permeability induced by AMPs is quantitatively the same in spheroplasts and GUVs. There are, however, interesting dissimilarities in details that reveal differences between bacterial and lipid membranes. Spheroplast membranes are permeabilized by a wide range of AMP concentrations to the same steady-state membrane permeability. In contrast, only a narrow range of AMP concentrations permeabilized GUVs to a steady state. Tension in GUVs also influences the action of AMPs, whereas the spheroplast membranes are tensionless. Despite these differences, our results provide a strong support for using model membranes to study the molecular interactions of AMPs with bacterial membranes. As far as we know, this is the first time the actions of AMPs, on bacterial membranes and on model membranes, have been directly and quantitatively compared.

Antenatal diagnosis of idiopathic dilatation of pulmonary artery with 3D power doppler imaging.

We present a case of antenatal diagnosis of idiopathic dilatation of the pulmonary artery by three-dimensional power Doppler imaging (HDliveFlow with silhouette mode). Two-dimensional sonography clearly demonstrated a 10.4-mm-diameter fetal pulmonary artery (PA), whereas the aorta (Ao) diameter was 5.04 mm. HDliveFlow clearly demonstrated the spatial relationships and different sizes of PA and Ao. The PA size returned to normal (8.0 mm) on the 11th day after birth (Ao: 9.0 mm). HDliveFlow may be an adjunctive tool to two-dimensional sonography to diagnose abnormalities of fetal great vessels. © 2016 Wiley Periodicals, Inc. J Clin Ultrasound 45:121-123, 2017.

Increased postpartum maternal complications after cesarean section compared with vaginal delivery in 225 304 Taiwanese women.

OBJECTIVE: To compare the short-term maternal postpartum complications associated with cesarean section (CS), vaginal delivery (VD), repeated CS and vaginal birth after cesarean section (VBAC) in a large national sample. METHODS: This was a population-based study of the Taiwan National Health Insurance Research Database (NHIRD). Outcomes include post-discharge (2 weeks) urinary tract infection (p-UTI), complications of obstetrical surgical wounds (p-wound) and postpartum hemorrhage (p-hemorrhage). A logistic regression model with generalized estimating equations were utilized, and adjustments were made for maternal and hospital characteristics. RESULTS: The incidence of p-UTI was 0.79%. CS was associated with a significantly higher risk of p-UTI compared with VD (odds ratio [OR] 1.14; 95% confidence interval [CI], 1.003-1.29). The incidence of p-wound was 4.07%. CS and repeated CS were associated with a higher risk of p-wound compared with VD (OR 1.68; 95% CI, 1.28-2.21 and OR 1.64; 95% CI, 1.22-2.20, respectively). Age, maternal diseases and hospital and obstetrician volumes were associated with patient outcomes. CONCLUSIONS: Women with a delivery mode of CS have a higher risk of p-UTI and p-wound than women with VD. Maternal characteristics and hospital and obstetrician volumes may also influence postpartum outcomes.

Prenatal diagnosis of parapagus diprosopus dibrachius dipus twins with spina bifida in the first trimester using two- and three-dimensional ultrasound.

OBJECTIVE: Here, we report a case of parapagus diprosopus twins with spina bifida diagnosed in the first trimester of pregnancy using two-dimensional (2D) and three-dimensional (3D) ultrasound. CASE REPORT: A 28-year-old Taiwanese woman, gravid 1, para 0, visited our hospital due to an abnormal fetal head shape discovered by 2D ultrasound at 11-weeks gestation. Parapagus diprosopus twins with spina bifida were diagnosed after ultrasound examination. The characteristics of parapagus diprosopus twins are more illustrative in 3D ultrasound than in 2D ultrasound. After counseling, termination of pregnancy was chosen by the couple. Although necropsy was declined, the gross appearance and radiograph of the abortus confirmed our diagnosis. CONCLUSION: With the help of 3D ultrasound, we made an early and definitive diagnosis of conjoined twins.

Three-dimensional ultrasonography measurement of fetal nasal bone length during the midtrimester in Taiwanese women.

OBJECTIVE: To evaluate the normal range of the fetal nasal bone length (NBL) in Taiwanese women using three-dimensional (3D) ultrasound, and compare the NBL of normal fetuses with Down syndrome to determine its significance in screening for trisomy 21. MATERIALS AND METHODS: A total of 102 consecutive fetuses and another 7 fetuses with trisomy 21, determined by karyotyping at 15-22 weeks' gestation, were evaluated with 3D ultrasound before amniocentesis at Changhua Christian Hospital between November 2003 and April 2004. RESULTS: The normal range for NBL in the second trimester in the Taiwanese population was investigated, and a linear relationship with gestational age was noted. The NBL increased with advancing gestational age (NBL in cm=0.0264×gestational age in weeks -0.042 (R(2)=0.2416). The median of the biparietal diameter/nasal bone length ratio had a stable value which tended to change minimally between 15 and 22 weeks of gestation. Chromosomally normal fetuses had statistically longer nasal bones than fetuses with Down syndrome (p=0.014). CONCLUSION: We present a reference range for 3D ultrasound measurement of the fetal NBL. A short nasal bone at 15 to 22 weeks is associated with a high risk of trisomy 21.

HepG2 cells infected with Klebsiella pneumoniae show DNA laddering at apoptotic and necrotic stages.

Virulent Klebasiella pneumoniae (KP) inflicts severe liver abscesses in infected patients. This study investigated how the bacterial infection affected cell survival at the molecular level, in a cultured cell model. A strain of KP highly virulent in mice was isolated from a patient with liver abscess, and was used to infect HepG2 cells. The infected cells were examined for their viability, DNA fragmentation, and proteins involved in apoptosis or necrosis. We found that the infection decreased the viability of HepG2 cells at 4 hours (h) to 12 h post infection (pi). DNA ladders appeared 6-16 h pi and flow cytometry analysis showed apoptosis at 3-5 h pi, secondary necrosis at 6-9 h pi and primary necrosis at 8-9 h pi. Cleavages of Caspase 7, Caspase 9, α-Fodrin, and PARP were evident at 2-4 h pi. At 7 h pi, we observed the following: increased nuclear AIF, the release from mitochondria of cytosolic Apaf-1 and Cyt c, increased DFF40 expression, decreased DFF45, decreased BcL-xL and the release of Endo G from mitochondria to nucleus. Cellular ATP concentration decreased at 4-8 h pi, accompanied by increased Calpain-2 expression. In summary, infected HepG2 cells underwent apoptosis early after infection and progressed to secondary necrosis and primary necrosis. Nuclear fragmentation corresponded to Caspase 7 activation and the appearance of Endo G and DFF40 in the nucleus, with a concomitant decrease in DFF45. Mitochondrial release of Cyt c together with activation of Caspase 9 and Apaf-1 in cytosol was also observed. Early-hour cleavage of poly(ATP-ribose) polymerase (PARP) followed by the later activation of PARP corresponded to the appearance of DNA laddering, and the depletion of cellular ATP was associated with the appearance of necrosis.

Prenatal diagnosis of persistent right umbilical vein using three-dimensional sonography with power Doppler.

OBJECTIVE: To investigate the incidence and the importance of isolated persistent right umbilical vein (PRUV) in our obstetric population and to determine the role of three-dimensional (3D) ultrasound in prenatal diagnosis of isolated PRUV. MATERIAL AND METHODS: A total of 1,302 women who received regular antenatal care by a sole obstetrician at our hospital were prospectively evaluated between July 2003 and April 2005. Detailed anatomical evaluation of the fetus was performed by one sonographer. When the diagnosis of PRUV was made, it was confirmed by a senior obstetrician. 3D ultrasound with power Doppler was applied to delineate local anatomy. Echocardiography was performed in all the newborns by pediatric cardiologists to confirm the prenatal diagnosis and to evaluate for the presence of associated anomalies. RESULTS: Six fetuses with PRUV were detected among the 1,302 study subjects. The incidence of PRUV in our population was 0.46% (1:217 live births). Vascular anatomy was easy to demonstrate using 3D power Doppler. The ductus venous (DV) was present in all six fetuses. An atrial septal defect was shown to exist in four newborns by neonatal echocardiography, but spontaneous closure had occurred in the follow-up scan. CONCLUSION: PRUV is a common vascular anomaly that is easy to be overlooked. Reconstruction of the portal system in the affected fetuses using 3D ultrasound facilitated the identification of the DV. If the DV is present, and other anomalies are excluded, the fetus with PRUV has a good outcome.

The detection of ureteral injuries after hysterectomy.

STUDY OBJECTIVE: To identify the incidence and outcomes of ureteral injuries in patients undergoing hysterectomy, and to evaluate the effect of intraoperative cystoscopy and early postoperative ureteral jet ultrasonography. DESIGN: Retrospective comparative study (Canadian Task Force classification II-2). SETTING: Tertiary medical center. PATIENTS: Fifteen patients who experienced ureteral injuries while undergoing hysterectomy out of 4950 total patients during a 6-year period. INTERVENTIONS: All patients underwent hysterectomy, and intraoperative cystoscopy or early postoperative ureteral jet ultrasonography were used to evaluate ureteral integrity. MEASUREMENTS AND MAIN RESULTS: The incidence of ureteral injury recognized after hysterectomy was 0.32%, and all ureteral injuries occurred during laparoscopic hysterectomy. The initial detection of ureteral injury resulted from intraoperative cystoscopy in five patients, early postoperative ureteral jet ultrasonography in two patients, and signs and symptoms in eight patients. The patients whose injuries were detected by either intraoperative cystoscopy or early postoperative ureteral jet ultrasonography were diagnosed earlier (1.7 and 19.9 postoperative days, respectively; p<.01) and tended toward more conservative treatment (p=.119; OR=10; 95% CI 0.78-128.78) than those who were diagnosed based on signs and symptoms alone. CONCLUSIONS: Use of intraoperative cystoscopy or early postoperative ureteral jet ultrasonography leads to earlier diagnosis of posthysterectomy ureteral injury, thereby allowing for more conservative treatment.