Donor cell engineering with GSK3 inhibitor-loaded nanoparticles enhances engraftment after in utero transplantation.

Host cell competition is a major barrier to engraftment after in utero hematopoietic cell transplantation (IUHCT). Here we describe a cell-engineering strategy using glycogen synthase kinase-3 (GSK3) inhibitor-loaded nanoparticles conjugated to the surface of donor hematopoietic cells to enhance their proliferation kinetics and ability to compete against their fetal host equivalents. With this approach, we achieved remarkable levels of stable, long-term hematopoietic engraftment for up to 24 weeks post-IUHCT. We also show that the salutary effects of the nanoparticle-released GSK3 inhibitor are specific to donor progenitor/stem cells and achieved by a pseudoautocrine mechanism. These results establish that IUHCT of hematopoietic cells decorated with GSK3 inhibitor-loaded nanoparticles can produce therapeutic levels of long-term engraftment and could therefore allow single-step prenatal treatment of congenital hematological disorders.

Influence of Lactobacillus crispatus, Lactobacillus iners and Gardnerella vaginalis on bacterial vaginal composition in pregnant women.

PURPOSE: To investigate associations between bacterial species in the vagina in mid-trimester pregnant women from Brazil. METHODS: The vaginal microbiome in 613 subjects was identified by analysis of the V1-V3 region of bacterial 16S ribosomal RNA and the relative prevalence of individual bacteria were determined. RESULTS: The bacterial species present in the greatest number of women were Lactobacillus crispatus (306 women), L. iners (298 women) and Gardnerella vaginalis (179 women). When present in the vagina, L. crispatus was the most abundant bacterium more than 85% of the time. In contrast, L. iners and G. vaginalis were most abundant in 63% and 41% of women who were positive for these microorganisms, respectively (p < 0.0001 vs. L. crispatus). The proportion of L. crispatus was negatively associated with the proportions of L. iners, L. jensenii, L. gasseri, G. vaginalis, Megasphaera, Atopobium vaginae and Prevotella (p < 0.0001). In contrast, the proportion of G. vaginalis was positively associated with levels of Megasphaera, A. vaginae and Prevotella (p < 0.0001) while L. iners proportion was unrelated to the proportion of L. jensenii, G. vaginalis, Megasphaera, A. vaginae or Prevotella. CONCLUSION: The composition of the vaginal microbiota in mid-trimester pregnant women is influenced by the relative concentrations of L. crispatus, L. iners and G. vaginalis.

Screening of Congenital Heart Diseases by Three-Dimensional Ultrasound Using Spatiotemporal Image Correlation: Influence of Professional Experience.

OBJECTIVE: To evaluate the feasibility of the use of spatiotemporal image correlation (STIC) as a screening program for congenital heart disease and the influence of professional experience in those examinations. METHODS: This prospective cross-sectional study included 64 pregnant women at gestational age between 20 and 34 weeks, and 12 physician participants who were divided into two groups: group 1-"STIC specialist"; and group 2-"STIC nonspecialist." Volumes were analyzed to obtain the five axial views for optimal fetal heart screening: abdominal situs, four-chamber view (4CV), outflow tract views (OTV), and three vessels and trachea view (3VT). The chi-square test (χ(2) ) was used to compare the group's results and kappa coefficient to evaluate inter- and intra-observer reproducibility. RESULTS: Spatiotemporal image correlation volume acquisition was successful in 97.3% of cases in which it was attempted (group 1: 100%; group 2:95%). A total of 197 STIC volumes were used in this study. In 71%, it was possible to demonstrate 4CV and OTV (group 1:88.9%; group 2:58.6%). 4CV, OTV, and 3VT were visualized in 55.3% of the volume dataset (group 1:74.1% and group 2:42.2%). In 49% of volumes, all five views for optimal fetal heart screening were seen (group 1:67%; group 2: 36%). A good inter-observer agreement was found in all cardiac views and a good intra-observer agreement in most of views except in OTV. CONCLUSION: We believe that STIC can be used as a tool to improve the cardiac screening examination of the fetus. Professional experience was the most important influence in the image quality of the STIC volume.

Characterization of Torquetenovirus in amniotic fluid at the time of in utero fetal surgery: correlation with early premature delivery and respiratory distress.

Torquetenovirus (TTV) is a commensal virus present in many healthy individuals. Although considered to be non-pathogenic, its presence and titer have been shown to be indicative of altered immune status in individuals with chronic infections or following allogeneic transplantations. We evaluated if TTV was present in amniotic fluid (AF) at the time of in utero surgery to correct a fetal neurological defect, and whether its detection was predictive of adverse post-surgical parameters. AF was collected from 27 women by needle aspiration prior to a uterine incision. TTV titer in the AF was measured by isolation of viral DNA followed by gene amplification and analysis. The TTV genomes were further characterized and sequenced by metagenomics. Pregnancy outcome parameters were subsequently obtained by chart review. Three of the AFs (11.1%) were positive for TTV at 3.36, 4.16, and 4.19 log10 copies/mL. Analysis of their genomes revealed DNA sequences similar to previously identified TTV isolates. Mean gestational age at delivery was >2 weeks earlier (32.5 vs. 34.6 weeks) and the prevalence of respiratory distress was greater (100% vs. 20.8%) in the TTV-positive pregnancies. TTV detection in AF prior to intrauterine surgery may indicate elevated post-surgical risk for earlier delivery and newborn respiratory distress.

The vaginal Torquetenovirus titer varies with vaginal microbiota composition in pregnant women.

Torquetenovirus (TTV) is a nonpathogenic endogenous virus whose abundance varies with the extent of immune system activation. We determined if the TTV titer in the vagina of pregnant women was associated with vaginal microbiota composition and levels of compounds in vaginal secretions. Vaginal TTV and microbiota composition in 494 second trimester pregnant women were identified by gene amplification and analysis. Vaginal matrix metalloproteinases (MMPs), tissue inhibitors of MMP (TIMP) and lactic acid isomers were measured by ELISA. Dominance was defined as the relative abundance of a specific bacterium or species at >50% of the total number of bacteria identified. Clinical data were obtained by chart review. The median log10 TTV titer was lowest when Lactobacillus species other than L. iners were dominant (<1.0) as compared to when L. iners (4.1, p = 0.0001), bacteria other than lactobacilli (4.5, p = 0.0016) or no bacterium (4.7, p = 0.0009) dominated. The TTV titer was inversely proportional to L. crispatus abundance (p<0.0001) and directly proportional to levels of G. vaginalis (p = 0.0008) and L. iners (p = 0.0010). The TTV titer was proportional to TIMP-1, TIMP-2, MMP-8 and MMP-9 abundance (p≤0.0002) and inversely proportional to the level of D-lactic acid (p = 0.0024). We conclude that the association between variations in the TTV titer and the relative abundance of specific bacterial species and vaginal compounds indicates that local changes in immune status likely influence vaginal fluid composition.

Histological response and expression of collagen, metalloproteinases MMP-1 and MMP-9 and tissue inhibitors of metalloproteinases TIMP-1 and TIMP-2 in fetal membranes following open intrauterine surgery: an experimental study.

OBJECTIVE: To characterize aspects of the repair process by evaluating the tissue collagen density, metalloproteinases and tissue inhibitor of matrix metalloproteinases in the fetal membranes following open fetal surgery for myelomeningocele (MMC). DESIGN: Experimental. SETTING: Two Brazilian hospitals in 2013-2014. POPULATION: 30 fetal membranes collected after elective cesarean deliveries, in patients who underwent open fetal surgery for MMC intrauterine repair. METHODS: Regions within and surrounding the scar area and regions distant from the surgical site were evaluated for collagen concentration and expression of MMP-1, MMP-9, TIMP-1 and TIMP-2. RESULTS: Collagen was increased in regions of scar formation (14.4 ± 2.7%) as compared to unaffected regions (8.0 ± 1.9%) (p < .001). The mean score of MMP-9 in the area of both the scar and suture was also increased above that observed in normal regions (p < .05). Conversely, MMP-1 was reduced in the scar when compared to the normal region and the area adjacent to the scar (suture region) (p < .05). TIMP-1 was increased in the suture region compared to the normal region (p < .05) while TIMP-2 was reduced in the scar region when compared to the other two regions (p < .05). The membrane repair process was also influenced by the number of previous pregnancies and gestational age at the time of surgery. CONCLUSION: Reparative activity of the fetal membrane after open fetal surgery involves up-regulation of collagen production and differential involvement of MMPs and TIMPs.

An exploratory study of associations with spontaneous preterm birth in primigravid pregnant women with a normal cervical length.

BACKGROUND: Predictors of spontaneous preterm birth in primigravid women remain undetermined. AIM: We evaluated whether biomarkers in vaginal secretions and/or differences in the dominant bacterium in the vaginal microbiome predicted the risk for spontaneous preterm birth in primigravid women with a cervical length >25mm. STUDY DESIGN: In a prospective study, 146 second trimester pregnant women with their first conception and a cervix >25mm were enrolled. The vaginal microbiome composition was characterized by analysis of 16S ribosomal RNA gene sequences. The concentrations of d- and l-lactic acid, matrix metalloproteinase (MMP) 2, 8 and 9 and tissue inhibitor of metalloproteinase (TIMP) 1 and 2 in vaginal secretions were measured by ELISA. Cervical length was determined by vaginal ultrasonography. Pregnancy outcome data were subsequently collected. There was a spontaneous preterm birth (SPTB) in 13 women (8.9%) while in an additional 8 women (5.5%) preterm delivery was medically indicated. Lactobacillus iners was the dominant vaginal bacterium in 61.5% of women with a SPTB but only in 31.2% of those who delivered at term (p = .0354). The vaginal concentration of TIMP-1 (p = .0419) and L-lactic acid (p = .0495) was higher in women with a SPTB as compared to those who delivered at term. Lactobacillus iners dominance was associated with elevated levels of TIMP-1 (p = .0434) and TIMP-2 (p = .0161) and lower levels of D-lactic acid (p < .0001) compared to when L. crispatus was dominant. CONCLUSION: In this exploratory study of primigravid women, elevations in vaginal TIMP-1 and L- lactic acid and L. iners dominance in the vaginal microbiome are associated with an increased occurrence of SPTB.

Reproducibility of fetal heart volume by 3D-sonography using the XI VOCAL method.

BACKGROUND: To assess the reliability of fetal heart volume measurement by three-dimensional sonography (3DUS) using the eXtended Imaging Virtual Organ Computer-aided AnaLysis (XI VOCAL) method. METHODS: This reliability study enrolled 30 pregnant women with singleton healthy pregnancies between 19 and 34 weeks of gestation. All volume acquirements were performed with a convex volumetric transducer (C3-7ED) coupled to an Accuvix XQ sonography device (Medison, Korea). The XI VOCAL 10 planes was the method of choice for volumetric measurement. 3D datasets were analyzed by two observers (EQSB and HJFM); fetal heart volume was measured twice by the first and once by the second observer to calculate intra and interobserver reproducibility. Statistical analysis used pareated Student's t test (p) and calculated Intraclass correlation coefficients (ICC). Bland-Altman plots were also constructed. RESULTS: We observed an excellent intra- and interobserver reliability for fetal cardiac volume assessed by XI VOCAL. For the intraobserver the ICC was 0.998 (95% CI: 0.997; 0.999), with mean of differences of 0.12 cm3 (95% limits of agreement: -0.84; +0.84; p = 0.130). For interobserver the ICC was 0.899 (95%CI: 0.996; 0.998), mean of differences 0.05 cm3 (95% limits of agreement: -0.84; +0.84; p = 0.175). CONCLUSION: Fetal cardiac volume assessed by 3DUS using XI VOCAL method is highly reproducible between 19 to 34 gestational weeks.

Transvaginal grey scale histogram of the cervix at 20-25 weeks of pregnancy.

AIM: To assess the echogenicity of the cervical stroma and area surrounding the cervical canal (cervical gland area - CGA) using grey scale histogram (GSH) in pregnancies between 20 and 25 weeks. METHODS: Cross-sectional study involving 149 pregnancies in the second trimester. Transvaginal sonography was performed to measure cervical length (biometric evaluation), assess the CGA and detect cervical funnelling (morphologic evaluation). The GSH was used for the objective assessment of the cervical stroma and CGA in the mid portion of the cervix. Mean, minimum, maximum and standard deviation measurements were obtained on the GSH and the CGA/stroma ratio was calculated. Intra-observer and inter-observer reproducibility were assessed using the intraclass correlation coefficient (ICC). RESULTS: Mean cervical length was 42.37 ± 7.05 mm (range 12-54 mm). Funnelling was detected in four women (2.7%) and one of these also had absent CGA. The mean GSH stroma value was 103 ± 25 (53-160) and the mean CGA value was 64.5 ± 23 (13-167). The mean CGA/stroma ratio was 0.62 ± 0.14 (0.24-1.29). Intra- and inter-observer reproducibility was excellent with ICC of 0.975 and 0.922 respectively. CONCLUSION: There are large differences in the texture of the cervical stroma and CGA on GSH allowing the objective differentiation of the two areas. The CGA/stroma ratio allows the objective assessment of the absence or presence of the CGA.

Contribution of Epithelial Cells to Defense Mechanisms in the Human Vagina.

PURPOSE OF REVIEW: The vaginal milieu in women differs from that of other mammals, including non-human primates, in composition of secretions, the endogenous microbiota, and level of acidity. These changes apparently reflect evolutionary variations that maximized productive responses to a uniquely human vaginal environment. This review will highlight recent findings on properties of human vaginal epithelial cells that contribute to maintenance of a healthy vaginal environment. RECENT FINDINGS: Vaginal epithelial cells are responsive to the composition of the vaginal microbiome even in women who are in apparently good health and do not exhibit any adverse physical symptoms. This is especially important during pregnancy when immune defenses are modified and an effective epithelial cell-derived anti-microbial activity is essential to prevent the migration to the uterus of bacteria potentially harmful to pregnancy progression. When Lactobacillus crispatus numerically predominates in the vagina, epithelial cell activity is low. Conversely, predominance of Lactobacillus iners, Gardnerella vaginalis, or other non-Lactobacilli evokes production and release of a large variety of compounds to minimize the potentially negative consequences of an altered microbiome. The extent of autophagy in vaginal epithelial cells, a basic process that functions to maintain intracellular homeostasis and engulf microbial invaders, is also sensitive to the external microbial environment Vaginal epithelial cells bind and release norepinephrine and upregulate their anti-microbial activity in response to external stress. Vaginal epithelial cells in women are responsive to local conditions that are unique to humans and, thereby, contribute to maintenance of a healthy milieu.

Vaginal Biomarkers That Predict Cervical Length and Dominant Bacteria in the Vaginal Microbiomes of Pregnant Women.

In many impoverished regions of the world, it may not be possible to assess two major risk factors for preterm birth: a short cervical length and the depletion of vaginal lactobacilli. We determined whether measuring specific compounds in vaginal fluid might be a simple, noninvasive, and cost-effective way to predict the bacteria that dominate the vaginal microbiome and indicate the presence of a shortened cervix (<25 mm). Vaginal fluid samples were prospectively collected from mid-trimester pregnant women, and the concentrations of d- and l-lactic acid, tissue inhibitor of matrix metalloproteinases TIMP-1 and TIMP-2, matrix metalloproteinases MMP-2 and MMP-8, the 70-kDa heat shock protein, a2 isoform of vacuolar ATPase, and sequestrome-1 were quantified by an enzyme-linked immunosorbent assay (ELISA). The compositions of vaginal microbiomes were assessed by analysis of the V1-V3 regions of 16S rRNA genes, while cervical length was determined by transvaginal ultrasonography. The vaginal microbiomes could be clustered into five community state types (CSTs), four of which were dominated by a single Lactobacillus species. The dominance of Lactobacillus crispatus or Lactobacillus jensenii in the vaginal microbiome predicted the level of d-lactic acid present. Several of the biomarkers, especially TIMP-1, in combination with the subject's age and race, were significantly associated with cervical length. Using piecewise structural equation modeling, we established a causal network that links CST to cervical length via biomarkers. We concluded that measuring levels of TIMP-1 and d-lactic acid in vaginal secretions might be a straightforward way to assess the risk for preterm birth due to a short cervix and microbiome composition.IMPORTANCE Premature birth and its complications are the largest contributors to infant death in the United States and globally. A short cervical length and the depletion of Lactobacillus species are known risk factors for preterm birth. However, in many resource-poor areas of the world, the technology to test for their occurrence is unavailable, and pregnant women with these risk factors are neither identified nor treated. In this study, we used path analysis to gain an unprecedented understanding of interactions between vaginal microbiome composition, the concentrations of various compounds in vaginal secretions, and cervical length. We identified low-cost point-of-care measures that might be used to identify pregnant women at risk for preterm birth. The use of these measures coupled with appropriate preventative or treatment strategies could reduce the incidence of preterm births in poor areas of the world that lack access to more sophisticated diagnostic methods.

Development of Secondary Microcephaly After Delivery: Possible Consequence of Mother-Baby Transmission of Zika Virus in Breast Milk.

BACKGROUND The Zika virus is an arbovirus that has as main source of transmission the bite of infected insects of the genus Aedes and has been associated with cases of congenital malformation and microcephaly in neonates. However, other sources of transmission have been identified since the emergence of this virus in the world population, such as vertical transmission by semen and possibly other body fluids such as vaginal secretion and breast milk. CASE REPORT An infant, born to a mother whose previous delivery was a baby with severe microcephaly, was normal and was negative for Zika virus at birth but developed secondary microcephaly 1 month later, that persisted. The baby was exclusively breast-fed and Zika virus was present in the mother's milk. CONCLUSIONS We report the detection of Zika virus exclusively in the breast milk of a woman after her second delivery of an infant, who later developed microcephaly. This case is consistent with possible vertical transmission.

Ophthalmic artery Doppler velocimetry in healthy pregnancy.

OBJECTIVES: To establish normative data for ophthalmic artery Doppler parameters throughout a healthy pregnancy, to investigate the possible correlation between these Doppler measurements and gestational age, and to test intraobserver variability. METHODS: A prospective cross-sectional study was performed to assess various Doppler parameters in 276 healthy women between 20 and 39 weeks of gestation. The following measurements were obtained: pulsatility and resistance indices (PI, RI), peak systolic velocity (PSV), peak diastolic velocity (PDV), end diastolic flow velocity (EDFV), and peak ratio (PR). Quantile regression was used to estimate reference values throughout pregnancy and P<0.05 was considered significant. The intraclass correlation coefficient (Intra-CC) was used to calculate intraobserver variability. RESULTS: The 5th and 95th prediction intervals for each gestational age are presented. No significant correlation was detected between Doppler indices and gestational age. Intra-CC was excellent (0.996). CONCLUSIONS: Reference values were generated for ophthalmic artery Doppler measurements, and they did not vary significantly during healthy pregnancy.

Antenatal corticosteroid treatment for the prevention of peri-intraventricular haemorrhage in preterm newborns: a retrospective cohort study using transfontanelle ultrasonography.

OBJECTIVE: The objective of this study was to assess the correlation between antenatal corticosteroids and peri-intraventricular haemorrhage (PIVH) using transfontanelle ultrasonography, as well as to evaluate the risk factors for its incidence. METHODS: We performed a retrospective cohort study using medical records of preterm newborns. The protocol for maternal corticoid administration for foetal lung maturation included dexamethasone 4 mg (intramuscular) 8/8 hours per 48 hours, with one cycle per week. The diagnosis of periintraventricular haemorrhage was based on transfontanelle ultrasonography, using the Papile's classification. The following risk factors for peri-intraventricular haemorrhage were assessed: birth weight, gestational age at delivery, type of delivery, newborn's sex, surfactant administration, premature rupture of membranes and previous history of infection during the current pregnancy. The student's t-test and chi-square test were used for statistical analysis. RESULTS: Our sample population included 184 preterm newborns. Transfontanelle ultrasonography revealed peri-intraventricular haemorrhage in 32 (74.4%) and periventricular leukomalacia in 11 (25.6%) newborns. Grade I haemorrhage was found in 20 (62.5%), grade II in five (15.6%), and grade III in seven (21.8%) newborns, as in accordance with Papile's classification. Vaginal delivery (p = 0.010), birth weight <1500 g (p = 0.024), gestational age at delivery ≤32 weeks (p = 0.018), and previous history of infection during pregnancy (p = 0.013) were considered risk factors for peri-intraventricular haemorrhage in preterm newborns. CONCLUSION: Maternal corticoid administration for foetal lung maturation showed a protective effect against peri-intraventricular haemorrhage in preterm newborns. The risk factors for peri-intraventricular haemorrhage were determined.

Histological evidence of reparative activity in chorioamniotic membrane following open fetal surgery for myelomeningocele.

An increased understanding of the reparative process in fetal membrane following surgical techniques may be helpful to decrease the risks to mother and fetus and avoid adverse pregnancy outcomes. The present study discusses histological evaluation of the fetal membrane following open fetal surgery. Chorioamniotic membranes (n=10) were obtained following birth from pregnancies that underwent open fetal surgery for myelomeningocele. The collagen distribution was quantified using picrosirius-polarization method comparing the suture site with non-suture site. The differences between the collagen fiber percentages at the two sites was evaluated by the paired t-test with P<0.05. The mean gestational age of fetal surgery was 26.09±0.3 and 33.81±0.82 weeks at birth. The picrosirius red sign was more intense at the suture site, primarily associated with collagen type 1. Collagen observed in the surgical area was significantly increased (13.22±2.84%) compared with the non-surgical area (6.16±1.09%; P<0.0001). It was observed that the reparative activity at the suture site of the fetal membrane was characterized by a significant increase in collagen fibers. The findings suggest nascent collagen synthesis, tissue remodeling and repair of suture site, a mechanism likely to prevent the amniotic fluid leakage and maintain pregnancy following open fetal surgery.

Maternal anxiety and fetal movement patterns in late pregnancy.

OBJECTIVE: To determine whether maternal anxiety affects fetal movement patterns in the third trimester of pregnancy. METHODS: The inclusion criteria were a state of good health and a singleton pregnancy between 36 and 40 weeks. Thirty healthy pregnant women were included. The Beck Anxiety Inventory (BA) questionnaire with 21 self-reported items validated for the Brazilian population was applied. The women were asked to record the number of minutes taken to perceive 10 fetal movements once a day for one week. Anxiety symptoms were rated as moderate or severe according to the BAI total score. RESULTS: The mean BAI score was 20.8 (SD = 10.2) and the mean time to count 10 fetal movements was 24.3 min (SD = 6.6 min). The BAI items significantly associated with moderate or severe maternal anxiety were numbness or tingling, fear of the worst happening, terrified, feeling of choking, fear of losing control and fear of dying. There was a statistically significant negative correlation between the total BAI score and the mean time of 10 perceived fetal movements (p < 0.0001; rho = -0.70; 95% CI for rho -0.84 to -0.45). CONCLUSIONS: Maternal anxiety seems to affect fetal movement patterns in late pregnancy and is associated with the mother's increased perception of fetal activity.

Antenatal diagnosis of renal duplication by ultrasonography: report on four cases at a referral center.

Duplication of the renal collecting system is the commonest major congenital malformation of the urinary tract, with an incidence of 1% among live births. Antenatal diagnosing of renal duplication and an associated ureterocele is infrequent. We report four cases of prenatally diagnosed unilateral duplication of the renal collecting system. In two of them, the renal duplication was associated with an ectopic ureterocele.

Lipidomic assessment of plasma and placenta of women with early-onset preeclampsia.

INTRODUCTION: Adipose tissue is responsible for triggering chronic systemic inflammatory response and these changes may be involved in the pathophysiology of preeclampsia. OBJECTIVE: To characterize the lipid profile in the placenta and plasma of patients with preeclampsia. METHODOLOGY: Samples were collected from placenta and plasma of 10 pregnant women with preeclampsia and 10 controls. Lipids were extracted using the Bligh-Dyer protocol and were analysed by MALDI TOF-TOF mass spectrometry. RESULTS: Approximately 200 lipid signals were quantified. The most prevalent lipid present in plasma of patients with preeclampsia was the main class Glycerophosphoserines-GP03 (PS) representing 52.30% of the total lipid composition, followed by the main classes Glycerophosphoethanolamines-GP02 (PEt), Glycerophosphocholines-GP01 (PC) and Flavanoids-PK12 (FLV), with 24.03%, 9.47% and 8.39% respectively. When compared to the control group, plasma samples of patients with preeclampsia showed an increase of PS (p<0.0001), PC (p<0.0001) and FLV (p<0.0001). Placental analysis of patients with preeclampsia, revealed the PS as the most prevalent lipid representing 56.28%, followed by the main class Macrolides/polyketides-PK04 with 32.77%, both with increased levels when compared with patients control group, PS (p<0.0001) and PK04 (p<0.0001). CONCLUSION: Lipids found in placenta and plasma from patients with preeclampsia differ from those of pregnant women in the control group. Further studies are needed to clarify if these changes are specific and a cause or consequence of preeclampsia.