Ascorbic acid supports ex vivo generation of plasmacytoid dendritic cells from circulating hematopoietic stem cells.

Plasmacytoid dendritic cells (pDCs) constitute a rare type of immune cell with multifaceted functions, but their potential use as a cell-based immunotherapy is challenged by the scarce cell numbers that can be extracted from blood. Here, we systematically investigate culture parameters for generating pDCs from hematopoietic stem and progenitor cells (HSPCs). Using optimized conditions combined with implementation of HSPC pre-expansion, we generate an average of 465 million HSPC-derived pDCs (HSPC-pDCs) starting from 100,000 cord blood-derived HSPCs. Furthermore, we demonstrate that such protocol allows HSPC-pDC generation from whole-blood HSPCs, and these cells display a pDC phenotype and function. Using GMP-compliant medium, we observe a remarkable loss of TLR7/9 responses, which is rescued by ascorbic acid supplementation. Ascorbic acid induces transcriptional signatures associated with pDC-specific innate immune pathways, suggesting an undescribed role of ascorbic acid for pDC functionality. This constitutes the first protocol for generating pDCs from whole blood and lays the foundation for investigating HSPC-pDCs for cell-based immunotherapy.

Vitamin D insufficiency among Danish pregnant women-Prevalence and association with adverse obstetric outcomes and placental vitamin D metabolism.

INTRODUCTION: In pregnancy, vitamin D deficiency is associated with increased risk of fetal growth restriction and preeclampsia. The underlying mechanisms are not known, but placental dysfunction is believed to play a role. In a Danish population, where health authorities recommend a 10 µg/day vitamin D supplement during pregnancy, we explored current use of vitamin D supplements and vitamin D status. In term placentas, alterations in vitamin D metabolism and placental growth, evaluated by the key placental growth factor pregnancy-associated plasma protein-A (PAPP-A), and their relation to vitamin D insufficiency were investigated. MATERIAL AND METHODS: We included 225 randomly selected pregnant women attending a nuchal translucency scan at gestational weeks 11-14. Information on use of vitamin D supplements and body mass index (BMI) at inclusion was obtained using self-reported questionnaires. Plasma 25-hydroxyvitamin D was measured at inclusion and correlated with pregnancy outcomes and placental biology, as judged by expression of PAPP-A and enzymes involved in vitamin D metabolism (CYP24A1, CYP27B1) in term placentas. RESULTS: Vitamin D supplements were used by 92% of the women, but 42% were vitamin D insufficient (plasma 25-hydroxyvitamin D <75 nmol/L). Eleven women with singleton pregnancies developed fetal growth restriction or preeclampsia. In this small subset, first-trimester mean plasma 25-hydroxyvitamin D was lower in women who developed fetal growth restriction (43 ± 33nmol/L; n = 3; P = .006) and there was a tendency towards lower plasma 25-hydroxyvitamin D among women who developed preeclampsia (65 ± 19 nmol/L; n = 8; P = .08) in third trimester compared with uncomplicated pregnancies (79 ± 22 nmol/L; n = 187). In term placentas, PAPP-A expression was lower among participants with first-trimester vitamin D insufficiency (P = .009; n = 30) but no correlation was found between plasma 25-hydroxyvitamin D and mRNA expression of CYP24A1 (P = .67) and CYP27B1 (P = .34). BMI was negatively correlated with plasma 25-hydroxyvitamin D (P = .03) and positively correlated with placental mRNA expression of CYP24A1 (P = .003; n = 30). CONCLUSIONS: Despite high compliance with official guidelines regarding vitamin D supplements, vitamin D insufficiency was frequent and the findings indicate that vitamin D insufficiency may affect placental growth. High BMI was associated with vitamin D insufficiency and increased placental vitamin D turnover, but further investigations are needed.

Effects of antenatal hypnosis on maternal salivary cortisol during childbirth and six weeks postpartum-A randomized controlled trial.

BACKGROUND: Cortisol has been used to capture psychophysiological stress during childbirth and postpartum wellbeing. We explored the effect of a brief antenatal training course in self-hypnosis on salivary cortisol during childbirth and 6 weeks postpartum. METHODS: In a randomized, controlled trial conducted at Aarhus University Hospital Skejby Denmark during the period January 2010 until October 2010, a total of 349 healthy nulliparous women were included. They were randomly allocated to a hypnosis group (n = 136) receiving three one-hour lessons in self-hypnosis with additional audio-recordings, a relaxation group (n = 134) receiving three one-hour lessons in various relaxation methods with audio-recordings for additional training, and a usual care group (n = 79) receiving ordinary antenatal care only. Salivary cortisol samples were collected during childbirth (at the beginning of the pushing state, 30 minutes, and 2 hours after childbirth), and 6 weeks postpartum (at wake up, 30 minutes after wake up, and evening). Cortisol concentrations were compared using a linear mixed-effects model. Correlations between cortisol concentrations and length of birth, experienced pain and calmness during birth were examined by a Spearman rank correlation test. FINDINGS: During childbirth, week correlations were found between cortisol concentrations 30 minutes after childbirth and length of birth. In the beginning of the pushing state and 2 hours after childbirth, we found a tendency towards higher cortisol concentrations in the hypnosis group compared to the other two groups (hypnosis versus relaxation p = 0.02 and 0.03, hypnosis versus usual care p = 0.08 and 0.05). No differences were observed in cortisol concentrations between the groups 30 minutes after childbirth (hypnosis versus relaxation p = 0.08, hypnosis versus usual care 0.10) or 6 weeks postpartum (hypnosis versus relaxation: p = 0.85, 0.51, and 0.68, hypnosis versus usual care: p = 0.85, 0.93, and 0.96). CONCLUSION: Antenatal hypnosis training may increase the release of cortisol during childbirth with no long-term consequences. Further research is needed to help interpret these findings.

Use of alternative medicine, ginger and licorice among Danish pregnant women - a prospective cohort study.

BACKGROUND: The use of alternative medicines and dietary supplements is constantly changing, as are dietary habits. One example of this phenomenon is the current popularity of ginger products as an everyday health boost. Ginger and licorice has also been shown to ameliorate nausea a common complaint in early pregnancy. Alternative medicines are often regarded as safe. However, they might affect fetal development, such as through alterations of hormone metabolism and cytochrome P450 function. Health care professionals may be unaware of the supplementation habits of pregnant women, which may allow adverse exposures to go unnoticed, especially if the rates of use in pregnancy are not known. We therefore investigated the use of alternative medicines and licorice among pregnant Danish women. METHODS: A total of 225 pregnant women were included in a prospective cohort when attending the national prenatal screening program at gestational weeks 10-16. Participants were asked to complete a questionnaire regarding their socio-economic status and lifestyle habits, including their intake of alternative medicine and licorice. RESULTS: We found that 22.7% of women reported taking alternative medicines, with 14.7% reporting daily consumption. Ginger supplements were consumed by 11.1%, mainly as health boost and 87.1% reported consumption of licorice. Regular or daily licorice consumption was reported by 38.2 and 7.1%, respectively. Notably, the use of licorice was reflected by an increase in blood pressure of the pregnant women. CONCLUSIONS: The use of licorice and alternative medicines appears to be common in pregnant Danish women, supporting the need for further investigations into the safety of alternative medicine use during pregnancy and the importance of up-to-date personalized counseling regarding popular health trends and lifestyle habits.

Women's Preferences for Birthing Hospital in Denmark: A Discrete Choice Experiment.

BACKGROUND: Free choice of hospital has been introduced in many healthcare systems to accommodate patient preferences and incentivize hospitals to compete; however, little is known about what patients actually prefer. OBJECTIVES: This study assessed women's preferences for birthing hospital in Denmark by quantifying the utility and trade-offs of hospital attributes. METHODS: We conducted a discrete-choice experiment survey with 12 hypothetical scenarios in which women had to choose between three hospitals characterized by five attributes: continuity of midwifery care, availability of a neonatal intensive care unit (NICU), hospital services offered, level of specialization to handle rare events, and travel time. A random parameter logit model was used to estimate the utility and marginal willingness to travel (WTT) for improvements in other hospital attributes. RESULTS: A total of 517 women completed the survey. Significant preferences were expressed for all attributes (p < 0.01), with the availability of a NICU being the most important driver of women's preferences; women were willing to travel 30 more minutes (95% confidence interval 28-32) to reach a hospital with a highly specialized NICU. The subgroup analyses revealed differences in WTT, with substantial heterogeneity due to prior experience with giving birth and regarding risk attitude and health literacy. CONCLUSION: A high specialization level was the most influential factor for women without previous birth experience and for risk-averse individuals but not for women with a high health literacy score. Hence, more information about the woman's risk profile and services required could play a role in affecting hospital choice.

Pregnant women's choice of birthing hospital: A qualitative study on individuals' preferences.

OBJECTIVE: To investigate pregnant women's decision making in relation to their choice of birthing hospital and, in particular, their priorities regarding hospital characteristics. METHODS: The focus of this study was the choice of birthing hospital among pregnant women. A qualitative interview design was used and women were recruited during their first pregnancy-related visit to a general practitioner. The interviews were conducted using a semi-structured interview guide, and a thematic analysis of the data was carried out. RESULTS: Women made their hospital choice decision independently and they relied extensively on their own or peers' experiences. Travel distance played a role, but some women were willing to incur longer travel times to give birth at a specialized hospital in order to try to reduce the risks (in case of unexpected events). The women associated the presence of specialized services and staff that were more qualified and experienced with increased safety. Other priorities included continuity of care (i.e., being seen by the same midwife) as well as service availability, which in this case referred to the possibility of a water birth and postnatal hoteling services. CONCLUSIONS: The choice of hospital provider appears to be strongly influenced by experience, whether personal experience or the experience of peers. However, there appears to be room for more information to be provided on safety and service attributes as an instrument for making an informed decision.

[Use of complementary and herbal medicine in the general population and among pregnant women].

Studies have reported a widespread use of herbal medicine in the general population (6-48%) and among pregnant women (4-69%) with great geographic and socio-economic variations in the extent of utilization and compounds used. The use of herbal medicine in Denmark remains relatively undescribed. Equivalent to conventional drugs, herbal medicine has side effects, interactions and contraindications. Thus, especially pregnant women should be careful as the safety profile remains unclear. Many patients do not report their use of herbal medicine to healthcare practitioners if they are not asked directly.

Antenatal hypnosis training and childbirth experience: a randomized controlled trial.

BACKGROUND: Childbirth is a demanding event in a woman's life. The aim of this study was to explore whether a brief intervention in the form of an antenatal course in self-hypnosis to ease childbirth could improve the childbirth experience. METHOD: In a randomized, controlled, single-blinded trial, 1,222 healthy nulliparous women were allocated to one of three groups during pregnancy: A hypnosis group participating in three 1-hour sessions teaching self-hypnosis to ease childbirth, a relaxation group receiving three 1-hour lessons in various relaxation methods and Mindfulness, and a usual care group receiving ordinary antenatal care only. Wijmas Delivery Expectancy/Experience Questionnaire (W-DEQ) was used to measure the childbirth experience 6 weeks postpartum. RESULTS: The intention-to-treat analysis indicated that women in the hypnosis group experienced their childbirth as better compared with the other two groups (mean W-DEQ score of 42.9 in the Hypnosis group, 47.2 in the Relaxation group, and 47.5 in the Care as usual group (p = 0.01)). The tendency toward a better childbirth experience in the hypnosis group was also seen in subgroup analyses for mode of delivery and for levels of fear. CONCLUSION: In this large randomized controlled trial, a brief course in self-hypnosis improved the women's childbirth experience.

Quantitative sonoelastography of the uterine cervix by interposition of a synthetic reference material.

OBJECTIVE: To develop a reference material that allows quantitative elastography of the uterine cervix using the calculation of the approximate tissue stiffness expressed as Young's modulus (N/mm(2) ). Further, to test the elastography equipment on phantoms from a clinical perspective regarding the distance dependence and the influence of a heterogeneous material. DESIGN: Methodological study. SETTING: Aarhus University Hospital, Denmark. POPULATION: Six mid- and five full-term pregnant women. METHODS: Reference caps and phantoms with Young's moduli between 0.07 and 0.40 N/mm(2) were made of silicone and oil. By using reference caps, the approximate Young's moduli of the cervixes were calculated from strain ratios obtained by elastography. MAIN OUTCOME MEASURES: Approximate Young's modulus of the cervix. RESULTS: The recordings of the phantoms revealed that the calculation of the approximate Young's moduli became unreliable at distances above 10-15 mm from the transducer. This was further increased for a phantom which included a soft layer imitating the cervical canal. The approximate Young's modulus obtained from the anterior cervical lip was 0.08 N/mm(2) in mid-term and 0.03 N/mm(2) in full-term pregnant women (Wilcoxon rank-sum test, p = 0.01). CONCLUSION: The reference cap constitutes a promising tool for quantitative elastography of the anterior cervical lip. Figures obtained from the posterior cervical lip are less plausible due to the distance from the transducer and the heterogeneity introduced by the cervical canal. The method has the potential to be used to supplement cervical length assessment when evaluating women at risk of preterm delivery and when planning induction of labor.

Effect of self-hypnosis on duration of labor and maternal and neonatal outcomes: a randomized controlled trial.

OBJECTIVE: To examine the effect of a brief course in self-hypnosis for childbirth on duration of the labor and other birth outcomes. DESIGN: A randomized, controlled, single-blind trial. SETTING: Aarhus University Hospital Skejby, Denmark. POPULATION: A total of 1222 healthy nulliparous women. METHODS: A hypnosis group receiving three 1-h lessons in self-hypnosis with additional audio-recordings to ease childbirth, a relaxation group receiving three 1-h lessons in various relaxation methods and mindfulness with audio-recordings for additional training, and a usual-care group receiving only the usual antenatal care were compared. MAIN OUTCOME MEASURES: Duration of labor, birth complications, lactation success, caring for the child, and preferred future mode of delivery. RESULTS: No differences were found across the three groups on duration from arriving at the birth department until the expulsive phase of second stage of labor, the duration of the expulsive phase, or other birth outcomes. Fewer emergency and more elective cesarean sections occurred in the hypnosis group. No difference was seen across the groups for lactation success or caring for the child but fewer women in the hypnosis group preferred a cesarean section in future pregnancies because of fear of childbirth and negative birth experiences. CONCLUSIONS: Learning self-hypnosis to ease childbirth taught as a brief course failed to show any effects on duration of childbirth and other birth outcomes.

Identification of circulating fetal cell markers by microarray analysis.

OBJECTIVE: Different fetal cell types have been found in the maternal blood during pregnancy in the past, but fetal cells are scarce, and the proportions of the different cell types are unclear. The objective of the present study was to identify specific fetal cell markers from fetal cells found in the maternal blood circulation at the end of the first trimester. METHOD: Twenty-three fetal cells were isolated from maternal blood by removing the red blood cells by lysis or combining this with removal of large proportions of maternal white blood cells by magnetic-activated cell sorting. Fetal cells identified by XY fluorescence in situ hybridization and confirmed by reverse-color fluorescence in situ hybridization were shot off microscope slides by laser capture microdissection. The expression pattern of a subset of expressed genes was compared between fetal cells and maternal blood cells using stem cell microarray analysis. RESULTS: Twenty-eight genes were identified as fetal cell marker candidates. CONCLUSION: Of the 28 fetal marker candidate genes, five coded for proteins, which are located on the outer surface of the cell membrane and not expressed in blood. The protein product of these five genes, MMP14, MCAM, KCNQ4, CLDN6, and F3, may be used as markers for fetal cell enrichment.