Effect of amniotic fluid stem cell transplantation on the recovery of bladder dysfunction in spinal cord-injured rats.

The effects of human amniotic fluid stem cell (hAFSC) transplantation on bladder function and molecular changes in spinal cord-injured (SCI) rats were investigated. Four groups were studied: sham and SCI plus phosphate-buffered saline (SCI + PBS), human embryonic kidney 293 (HEK293) cells, and hAFSCs transplantation. In SCI + PBS rat bladders, cystometry showed increased peak voiding pressure, voiding volume, bladder capacity, residual volume, and number of non-voiding contractions, and the total elastin/collagen amount was increased but collagen concentration was decreased at days 7 and 28. Immunoreactivity and mRNA levels of IGF-1, TGF-ß1, and ß3-adrenoceptor were increased at days 7 and/or 28. M2 immunoreactivity and M3 mRNA levels of muscarinic receptor were increased at day 7. M2 immunoreactivity was increased, but M2/M3 mRNA and M3 immunoreactivity levels were decreased at day 28. Brain derived-neurotrophic factor mRNA was increased, but immunoreactivity was decreased at day 7. HEK293 cell transplantation caused no difference compared to SCI + PBS group. hAFSCs co-localized with neural cell markers and expressed BDNF, TGF-ß1, GFAP, and IL-6. The present results showed that SCI bladders released IGF-1 and TGF-ß1 to stimulate elastin and collagen for bladder wall remodelling, and hAFSC transplantation improved these changes, which involved the mechanisms of BDNF, muscarinic receptors, and ß3-adrenoceptor expression.

Amniotic Fluid Stem Cells Improve Rat Bladder Dysfunction After Pelvic Nerve Transection.

The effects of human amniotic fluid stem cells (hAFSCs) transplantation on bladder dysfunction after pelvic nerve transection (PNT) remain to be clarified. Five groups of female Sprague-Dawley rats were studied including sham operation, unilateral PNT alone or plus hAFSCs transplantation, and bilateral PNT alone or plus hAFSCs transplantation. hAFSCs were injected at the site of PNT. Cystometries, neurofilament density within bladder nerves, and the expressions of bladder protein gene-product 9.5 (PGP9.5), growth-associated protein 43 (GAP-43), nerve growth factor (NGF), p75 (NGF receptor), CXCL12, CCL7, and enkephalin were studied. Compared to sham-operation group, bladder weight increased and neurofilament density decreased at 10 and 28 days after unilateral and bilateral PNT, but all improved after hAFSCs transplantation. Unilateral PNT could increase bladder capacity, residual volume, and number of nonvoiding contractions but decrease peak voiding pressure and leak point pressure. Bilateral PNT caused overflow incontinence and increased the number of nonvoiding contractions. These cystometric parameters improved after hAFSCs transplantation. After PNT, bladder PGP9.5 mRNA and immunoreactivities decreased at 10 and 28 days, GAP-43 mRNA and immunoreactivities increased at 10 days and decreased at 28 days, both NGF and p75 mRNAs and immunoreactivities increased at 10 and/or 28 days, and enkephalin immunoreactivities decreased at 10 and 28 days, but these were all improved after hAFSCs transplantation. Our results showed that bladder dysfunction induced by PNT could be improved by hAFSCs transplantation, and PGP9.5, GAP-43, and neurotrophins could be involved in the mechanisms of nerve regeneration after hAFSCs transplantation.

Detection of fetal trisomy 9 mosaicism by noninvasive prenatal testing through maternal plasma DNA sequencing.

OBJECTIVE: Noninvasive prenatal testing (NIPT) is widely used as a powerful screening tool to detect common aneuploidies. However, its application for detection of rare chromosomal abnormalities remains inconclusive. CASE REPORT: A 38-year-old woman (gravida 2, para 0) requested NIPT as a primary screening test for fetal aneuploidies at 13 weeks and 1 day of gestation. An unexpected Trisomy 9 (T9) abnormality was highly suspected. Amniocentesis was arranged for further diagnosis at 18 weeks of gestation. Final karyotyping reported 47,XX,+9 [18]/46,XX [12], indicating 60% T9 mosaicism. CONCLUSION: This case shows strong evidence that NIPT can be a powerful screening tool to detect rare fetal trisomies at very early gestation.

Prenatal transplantation of human amniotic fluid stem cells for spinal muscular atrophy.

PURPOSE OF REVIEW: To review the current medical and stem-cell therapy for spinal muscular atrophy (SMA) and prenatal transplantation of amniotic fluid stem cells in the future. RECENT FINDINGS: SMA is an autosomal recessive inheritance of neurodegenerative disease, which is caused of the mutation in survival motor neuron. The severe-type SMA patients usually die from the respiratory failure within 2 years after birth. Recently, researchers had found that 3-methyladenine could inhibit the autophagy and had the capacity to reduce death of the neurons. The first food and drug administration-approved drug to treat SMA, Nusinersen, is a modified antisense oligonucleotide to target intronic splicing silencer N1 just recently launched. Not only medical therapy, but also stem cells including neural stem cells, embryonic stem cells, mesenchymal stem cells, and induced pluripotent stem cells could show the potential to repair the injured tissue and differentiate into neuron cells to rescue the SMA animal models. Human amniotic fluid stem cells (HAFSCs) share the potential of mesenchymal stem cells and could differentiate into tri-lineage-relative cells, which are also having the ability to restore the injured neuro-muscular function. In this review, we further demonstrate the therapeutic effect of using HAFSCs to treat type III SMA prenatally. HAFSCs, similar to other stem cells, could also help the improvement of SMA with even longer survival. SUMMARY: The concept of prenatal stem-cell therapy preserves the time window to treat disease in utero with much less cell number. Stem cell alone might not be enough to correct or cure the SMA but could be applied as the additional therapy combined with antisense oligonucleotide in the future.

Prenatal diagnosis of fetal glutaric aciduria type 1 with rare compound heterozygous mutations in GCDH gene.

OBJECTIVE: Glutaric aciduria type 1 is a rare disease, with the estimated prevalence about 1 in 100,000 newborns. GCDH gene mutation can lead to glutaric acid and 3- OH glutaric acid accumulation, with clinical manifestation of neuronal damage, brain atrophy, microencephalic macrocephaly, decreased coordination of swallowing, poor muscle coordination, spasticity, and severe dystonic movement disorder. CASE REPORT: A 22-year-old female, Gravida 4 Para 2, is pregnancy at 13 weeks of gestational age. Her first child is normal, however, the second child was diagnosed as glutaric aciduria type I after birth. She came to our hospital for prenatal genetic counselling of her fetus at 13 weeks of gestational age. We performed GCDH gene mutation analysis of maternal blood showed IVS 3 + 1 G > A heterozygous mutation, GCDH gene mutation analysis of paternal blood showed c. 1240 G > A heterozygous mutation, and the second child has compound heterozygous IVS 3 + 1 G > A and c. 1240 G > A mutations. Later, we performed amniocentesis at 16 weeks of gestational age for chromosome study and GCDH gene mutation analysis for the fetus. The fetal chromosome study showed normal karyotype, however, GCDH gene mutation analysis showed compound heterozygous IVS 3 + 1 G > A and c. 1240 G > A mutations. The couple decided to termination of pregnancy thereafter. CONCLUSION: Glutaric acidemia type 1 is an autosomal recessive disorder because of pathogenic mutations in the GCDH gene. Early diagnosis and therapy of glutaric acidemia type 1 can reduce the risk of neuronal damage and acute dystonia. We report a case of prenatal diagnosis of fetal glutaric aciduria type 1 with rare compound heterozygous GCDH gene mutation at IVS 3 + 1 G > A and c. 1240 G > A mutations, which provide better genetic counselling for the couples.

Improvement in bladder dysfunction after bladder transplantation of amniotic fluid stem cells in diabetic rats.

To examine the effects of human amniotic fluid stem cells (hAFSCs) transplantation on bladder function and molecular changes in diabetic rats, 60 female Sprague-Dawley rats were used for study. Three groups were assigned including sham control rats, streptozotocin (STZ, 60 mg/kg)-induced diabetic rats and STZ-induced diabetic rats plus bladder hAFSCs transplantation. Compared to controls, diabetic rats had decreased body weight but increased bladder weight. Cystometries showed non-voiding contraction, residual volume, voided volume and intercontraction interval increased significantly in diabetic rats at week 4 and 12 after DM induction, but improved after hAFSCs transplantation. The immunoreactivities and mRNAs of nerve growth factor (NGF) decreased significantly in diabetic bladder at week 4 and 12 after DM induction, but recovered after hAFSCs transplantation. The immunoreactivities and mRNAs of M2 and M3 muscarinic receptor increased significantly in diabetic bladder at week 4 after DM induction but recovered after hAFSCs transplantation. The immunoreactivity of 8-hydroxy-20-deoxyguanosine increased significantly in diabetic bladder at week 4 and 12 after DM induction but reduced after hAFSCs transplantation. The present study showed bladder dysfunction in STZ-induced diabetic rats could be improved by hAFSCs transplantation into bladder, which may be related to the recovery of bladder NGF and muscarinic receptors.

Amniotic fluid stem cells ameliorate bladder dysfunction induced by chronic bladder ischemia in rat.

AIMS: This study investigated the protective effect of human amniotic fluid-derived stem cells (hAFSCs) against bladder overactivity in rat model of atherosclerosis-induced chronic bladder ischemia. METHODS: Adult female Sprague-Dawley rats were divided into six groups: (1) Normal control with a regular diet for 8 weeks, (2) Sham-operation, (3) arterial balloon endothelial injury (AEI) of common iliac artery (AEI only), and post-AEI consecutive hAFSCs treatment for (4) 1 day, (5) 3 days, and (6) 7 days. Groups 2-6 were given 2% cholesterol diet for 8 weeks after operation (sham or AEI). Bladder functions were analyzed by cystometry at 8 weeks in controls and after operation in groups 2-6. Wall morphology of common iliac artery was examined by hematoxylin and eosin stain. Bladder oxidative stress and inflammatory markers were studied by immunohistochemistry of 8-hydroxy-2'-deoxyguanosine (8OHdG), malondialdehyde (MDA), and tumor necrosis factor-alpha (TNF-alpha). RESULTS: Bladder overactivity with decreased voided volumes and intercontraction intervals and increased residual volumes was seen in AEI only group, but improved after hAFSCs treatment for 1, 3, and 7 days. Compared with controls and shams, the wall thickness of iliac artery was increased in AEI only group, but improved after hAFSCs treatment for 3 and 7 days. The expressions of 8OHdG, MDA, and TNF-alpha were increased in AEI only group, but improved after hAFSCs treatment for 1, 3, and 7 days. CONCLUSIONS: Bladder overactivity caused by chronic bladder ischemia can be improved by hAFSCs treatment, probably by acting through down-regulation of oxidative stress and TNF-alpha expressions.

Corrigendum: In utero therapy for congenital disorders using amniotic fluid stem cells.

[This corrects the article on p. 270 in vol. 5, PMID: 25566071.].

The impact of Down syndrome screening on Taiwanese Down syndrome births: a nationwide retrospective study and a screening result from a single medical centre.

A retrospective analysis of the Taiwanese National Birth Defect Registration and Notification System was conducted in order to determine the live birth- and stillbirth rates in infants with Down syndrome, trisomy 18, trisomy 13 and Turner syndrome between 2001 and 2010. The objective was to investigate the impact of Down syndrome screening on the Taiwanese Down syndrome live birth rate. In addition, the results of first-trimester Down syndrome screening between 2006 and 2011, and of second-trimester quadruple testing between 2008 and 2011, were obtained from the National Taiwan University Hospital. All Taiwanese infants born between 2001 and 2010 were included in the first part of the analysis, and women receiving first-trimester Down syndrome screening or second-trimester quadruple testing from the National Taiwan University Hospital were included in the second part. The live birth rate of infants with Down syndrome, per 100 000 live births, decreased from 22.28 in 2001 to 7.79 in 2010. The ratio of liveborn DS to total DS was 48.74% in 2001, and then decreased to 25.88% in 2006, when first-trimester screening was widely introduced in Taiwan. This ratio dropped to 20.64% in 2008, when the second-trimester quadruple test was implemented. The overall positive rate in first-trimester screening in the National Taiwan University Hospital was 3.1%, with a Down syndrome detection rate of 100%; the quadruple test had values of 9.0% and 75%, respectively. The use of first-trimester screening and the second-trimester quadruple test may be responsible for the marked decrease in the Taiwanese Down syndrome live birth rate observed between 2001 and 2010.

Factors associated with the intention to recommend pertussis vaccination for postpartum women: a survey in Taiwan of obstetrician-gynecologists' knowledge, beliefs, and attitudes.

OBJECTIVE: To examine obstetrician-gynecologists' knowledge, beliefs, and attitudes associated with the intention to recommend adult tetanus, reduced diphtheria, and acellular pertussis (Tdap) vaccination to postpartum women. MATERIALS AND METHODS: A survey instrument was mailed to a total of 2686 members of the Taiwan Association of Obstetrics and Gynecology to assess provider demographic characteristics, occupational information, pertussis knowledge, and beliefs and attitudes about vaccination. The intention to recommend pertussis vaccination to postpartum women was evaluated. Trend chi-square statistics and multivariate logistic models were used to determine variables that were significantly associated with intention to recommend vaccination. RESULTS: Of the 676 surveys returned (25.2%), 510 participants were active in obstetric practice. A statistically significant difference was found in mean ± SD knowledge scores for pertussis disease and pertussis vaccination between obstetrician-gynecologists who intended to recommend and those who did not intend to recommend postpartum Tdap vaccination (disease: 2.99 ± 2.2 vs. 2.31 ± 1.9, respectively, p < .001; vaccination: 2.64 ± 2.2 vs. 1.36 ± 1.8, respectively, p < .001). Obstetrician-gynecologists who were in favor of postpartum Tdap vaccination were more likely to: (1) rate the change in pertussis incidence among adults as increased; (2) rate pertussis disease among newborn infants as highly severe; (3) rate pertussis as highly contagious; and (4) understand the current recommendation of important organizations for protecting infants against pertussis. CONCLUSION: Our study of obstetrician-gynecologists' attitudes and intentions concerning postpartum Tdap vaccination may be useful in guiding the design of effective perinatal Tdap immunization programs nationwide.

Clinical applications of prenatal and postnatal therapy using stem cells retrieved from amniotic fluid.

PURPOSE OF REVIEW: To review the potential of stem cells derived from amniotic fluid and applications in prenatal and postnatal therapy. RECENT FINDINGS: We have recently described that pluripotent stem cells can be isolated from amniotic fluid defined as amniotic fluid stem (AFS) cells by selection for expression of the membrane stem cell factor receptor c-Kit. AFS cells maintained for over 250 population doublings retained long telomeres and normal karyotype. Clonal human lines verified by retroviral marking were induced to differentiate into cell types representing each embryonic germ layer, including adipogenic, osteogenic, myogenic, endothelial, neuronal, and hepatic lineages. Rat AFS cells have been able to improve the repair of damaged smooth muscle in cryoinjury bladders. Furthermore, AFS cells could be differentiated toward cardiomyogenic lineages, when co-cultured with neonatal cardiomyocytes and have potential to generate hematopoietic lineages both in vitro and in vivo. These cells have been applied into fetal therapy, and widely used for tissue repair in animal models. Finally, we demonstrated a feasible way to do in-utero autologous AFS transplantation in sheep. SUMMARY: Stem cells derived from amniotic fluid are a relatively new source of cells that could have a therapeutic value in various diseases prenatally and/or postnatally.

Maternal homocysteine level and markers used in first-trimester screening for fetal Down syndrome.

This study examined possible relationships between homocysteine and markers used in first-trimester screening for Down syndrome. Pregnancies were categorized into 4 groups according to quartile ranking of maternal plasma homocysteine concentration. Of the 595 pregnancies, 147 were assigned to group 1 (homocysteine level 0.6-3.5 µmol/L), 156 to group 2 (homocysteine level 3.6-4.5 µmol/L), 142 to group 3 (homocysteine level 4.6-5.6 µmol/L), and 150 pregnancies to group 4 (homocysteine level 5.7-12.6 µmol/L). No significant difference in mean nuchal translucency and mean free ß-human chorionic gonadotropin (free-ßhCG) multiples of the median (MoM) levels were observed. However, the mean pregnancy-associated plasma protein A (PAPP-A) MoM levels were significantly decreased in inverse relationship with homocysteine level among all 4 groups (F = 31.127, P < .001). If homocysteine is assayed as part of the first-trimester maternal serum testing, it is important to adjust for homocysteine concentration when using PAPP-A serum level for calculating the risk of fetal aneuploidy.

Evaluation of fetal spine biometry between 11 and 14 weeks of gestation.

This study was designed to establish a fetal spine nomogram for age 11 through 14 weeks of gestation and to document relations among fetal spine length, distance and angle. These parameters were prospectively measured during the first trimester of singleton pregnancies, along with nuchal translucency, over a 3-year period. A total of 430 fetuses were included in the study. The regression equations among fetal spine parameters and gestational age were as follows: Spine length (mm) = 1.116 x gestational age (days) - 59.169; spine distance (mm) = 1.079 x gestational age (days) - 59.038; head-spine angle = 0.740 x gestational age (days) + 4.735; spine length:spine distance ratio = -0.002 x gestational age (days) + 1.234. Prenatal age-specific reference intervals for fetal spine biometry between 11 and 14 weeks of gestation may assist in evaluation of fetuses investigated for genetic abnormalities that can be expressed by deviation in spine length, distance, or angle.

Factors influencing women's decisions regarding pertussis vaccine: A decision-making study in the Postpartum Pertussis Immunization Program of a teaching hospital in Taiwan.

We conducted surveys to determine factors influencing women's decisions to accept or decline postpartum pertussis (Tdap) vaccination. Survey response rate among eligible individuals was 97%. Of respondents, 53% accepted and 47% declined postpartum Tdap. Women, who declined vaccination were more likely to rate maternal or infant risk of exposure to pertussis as low, report that they did not trust information about postpartum pertussis vaccination, and report being very concerned about the safety of the vaccine. Awareness about pertussis, its risk to infants, and prevention via vaccination need to be further increased among women of child-bearing age, particularly pregnant women.

Intracardiac echogenic focus and trisomy 21 in a population previously evaluated by first-trimester combined screening.

OBJECTIVE: To determine the relation between intracardiac echogenic focus (ICEF) and trisomy 21 in a population of fetuses previously evaluated by first-trimester combined screening. DESIGN: Prospective study to evaluate the prevalence of trisomy 21 for four groups of pregnancies: those with isolated ICEF, those with ICEF and positive first-trimester combined screening, those with ICEF and other ultrasound markers, and those with ICEF, positive first-trimester combined screening and other ultrasound markers. SETTING: Teaching hospital. POPULATION: A total of 7,118 fetuses received first-trimester combined screening and a second-trimester fetal anatomical scan. METHODS: Amniocentesis for karyotyping was offered for those with positive result of first-trimester combined screening, and for those with the diagnosis of ICEF or other fetal chromosomal aneuploidy ultrasound markers in mid-trimester detailed fetal anatomical scans. MAIN OUTCOME MEASURES: The prevalence of trisomy 21 in each group. RESULTS: For the entire study population, 25 trisomy 21 cases were diagnosed prenatally (prevalence, 0.35%). Among fetuses with positive first-trimester combined screening results, the likelihood ratio of trisomy 21 was significantly increased for those with, as compared to without, ICEF (3.5 vs. 0.80, p = 0.012). However, among fetuses with negative first-trimester combined screening results, the likelihood ratios of trisomy 21 did not differ significantly between those with and without ICEF. CONCLUSION: In pregnancies complicated by isolated ICEF but lacking additional anomalies on ultrasonographic examination and exhibiting negative first-trimester combined screening results, fetal karyotyping is not indicated.

Effect of maternal hepatitis B carrier status on first-trimester markers of Down syndrome.

Up to 10% of women of reproductive age in our country are carriers of hepatitis B virus. This study examined whether maternal hepatitis B carrier status has any effect on markers used in first-trimester screening for Down syndrome. Records for 2 major Taiwanese hospitals were retrospectively examined to identify women with singleton pregnancies resulting in normal live births from June 2002 through 2008. Maternal hepatitis B data were used to define 3 groups: seronegative women, inactive carrier women, and active carrier women. Women with active or inactive carrier status were significantly older than seronegative women. The results of the study show that maternal hepatitis B carrier status does not influence first-trimester levels of maternal serum free beta-human chorionic gonadotropin (free beta-hCG) multiples of the median (MoM), pregnancy-associated plasma protein A (PAPP-A) MoM, and median fetal nuchal translucency and screening false-positive rate; therefore, correction in the risk calculation algorithm for maternal hepatitis B carrier status is not necessary.

Difference in nuchal translucency between monozygotic and dizygotic spontaneously conceived twins.

OBJECTIVE: To assess nuchal translucency (NT) thickness and differences in inter-twin NT thickness among naturally conceived monozygotic and dizygotic twins. METHODS: Enrolled women prospectively underwent NT measurement for both twins between 11 and 14 weeks of gestational age. NT thickness and inter-twin NT difference were recorded and compared among dizygotic twins, monozygotic dichorionic twins, and monozygotic monochorionic twins. RESULTS: This study obtained analyzable data for 174 monozygotic twins and 107 dizygotic twins. Of the 174 monozygotic twins, 105 were monochorionic, and 69 were dichorionic. Monozygotic monochorionic twin pregnancies had a significantly higher mean fetal NT thickness (1.48 +/- 0.62 mm) than either dizygotic twin pregnancies (1.34 +/- 0.50 mm) or monozygotic dichorionic twin pregnancies (1.31 +/- 0.48 mm). Mean inter-twin NT difference was significantly lower in monozygotic dichorionic twin pregnancies (0.37 +/- 0.19 mm) than in dizygotic twin pregnancies (0.57 +/- 0.22 mm) or monochorionic twin pregnancies (0.65 +/- 0.43 mm).Irrespective of twin zygosity, NT thickness and inter-twin NT difference were larger in monochorionic twins than in dichorionic twins (1.48 +/- 0.62 mm versus 1.33 +/- 0.49 mm, and 0.65 +/- 0.43 mm versus 0.49 +/- 0.23 mm, respectively). CONCLUSION: During first-trimester ultrasonographic screening for Down syndrome, monochorionic twin pregnancies have the greatest NT thickness and monozygotic dichorionic twin pregnancies have the lowest inter-twin NT difference.

Gene gun administration of therapeutic HPV DNA vaccination restores the efficacy of prolonged defrosted viral based vaccine.

Freshly defrosted vaccines generate promising antitumor immunity by raising both robust CD8 and CD4 responses with a TC1/Th1-dominant cytokine profile. However, prolonged (overnight) defrosted Sindbis virus-E7/HSP70 priming and Vaccinia-E7/HSP70 booster in mouse model only elicited 20% long-term tumor-free survival in comparison with the fresh vaccines. The present study is to search the possible cause of its potency loss, and to evaluate the ability of pcDNA-E7/HSP70 DNA vaccination via gene gun in restoring the efficacy of E7-specific immune responses and antitumor properties. We used prolonged defrosted SINrep5-E7/HSP70 prime and defrosted Vac-E7/HSP70 boost subcutaneously, and administered intradermally cluster (3-day interval) gene gun plasmid E7-HSP70DNA vaccine twice, and evaluated its ability to generate antigen-specific cytotoxic CD8+ T-cell responses using flow cytometry as well as antitumor responses using animal positron-emission tomography (PET) imaging. The prolonged defrosted vaccines showed a significant reduction in the infectivity and a significant decrease of CD8+ and CD4+ T-cells immune responses. Administration of cluster gene gun plasmid E7-HSP70DNA twice was also found to lead to restoration of immunity that elicits a full recovery of the antitumor efficacy of the prolonged defrosted vaccines. Our study suggested that adding cluster gene gun plasmid E7-HSP70DNA vaccine twice offered a simple solution in restoring the efficacy of the prime-boost vaccination with viral vectors and has potentially significant clinical applications.