Amniotic Fluid Concentrations Research Articles

Association of Fetal Catecholamines With Neonatal Hypoglycemia

Perinatal stress and fetal growth restriction increase the risk of neonatal hypoglycemia. The underlying pathomechanism is poorly understood. In a sheep model, elevated catecholamine concentrations were found to suppress intrauterine insulin secretion, followed by hyperresponsive insulin secretion once the adrenergic stimulus subsided. To determine whether neonates with risk factors for hypoglycemia have higher catecholamine concentrations in umbilical cord blood (UCB) and/or amniotic fluid (AF) and whether catecholamines are correlated with postnatal glycemia. In a prospective cohort study of 328 neonates at a tertiary perinatal center from September 2020 through May 2022 in which AF and UCB were collected immediately during and after delivery, catecholamines and metanephrines were analyzed using liquid chromatography with tandem mass spectrometry. Participants received postnatal blood glucose (BG) screenings. Risk factor for neonatal hypoglycemia. Comparison of catecholamine and metanephrine concentrations between at-risk neonates and control participants, and correlation of concentrations of catecholamines and metanephrines with the number and severity of postnatal hypoglycemic episodes. In this study of 328 neonates (234 in the risk group: median [IQR] gestational age, 270 [261-277] days; and 94 in the control group: median [IQR] gestational age, 273 [270-278] days), growth-restricted neonates showed increased UCB median (IQR) concentrations of norepinephrine (21.10 [9.15-42.33] vs 10.88 [5.78-18.03] nmol/L; P < .001), metanephrine (0.37 [0.13-1.36] vs 0.12 [0.08-0.28] nmol/L; P < .001), and 3-methoxytyramine (0.149 [0.098-0.208] vs 0.091 [0.063-0.149] nmol/L; P = .001). Neonates with perinatal stress had increased UCB median (IQR) concentrations of norepinephrine (22.55 [8.99-131.66] vs 10.88 [5.78-18.03] nmol/L; P = .001), normetanephrine (1.75 [1.16-4.93] vs 1.25 [0.86-2.56] nmol/L; P = .004), and 3-methoxytyramine (0.120 [0.085-0.228] vs 0.091 [0.063-0.149] nmol/L; P = .008) (P < .0083 was considered statistically significant). Concentrations of UCB norepinephrine, metanephrine, and 3-methoxytyramine were negatively correlated with AF C-peptide concentration (rs = -0.212, P = .005; rs = -0.182, P = .016; and rs = -0.183, P = .016, respectively [P < .017 was considered statistically significant]). Concentrations of UCB norepinephrine, metanephrine, and 3-methoxytyramine were positively correlated with the number of hypoglycemic episodes (BG concentration of 30-45 mg/dL) (rs = 0.146, P = .01; rs = 0.151, P = .009; and rs = 0.180, P = .002, respectively). Concentrations of UCB metanephrine and 3-methoxytyramine were negatively correlated with the lowest measured BG concentration (rs = -0.149, P = .01; and rs = -0.153, P = .008, respectively). Neonates at risk for hypoglycemia displayed increased catecholamine and metanephrine concentrations that were correlated with postnatal hypoglycemic episodes and lower BG levels; these results are consistent with findings in a sheep model that fetal catecholamines are associated with neonatal β-cell physiology and that perinatal stress or growth restriction is associated with subsequent neonatal hyperinsulinemic hypoglycemia. Improving the pathomechanistic understanding of neonatal hypoglycemia may help to guide management of newborns at risk for hypoglycemia.

Clinical application of amino-terminal pro-brain natriuretic peptide concentration in amniotic fluid for the prediction of preterm birth in symptomatic women

BackgroundPreterm birth accounts for 60-80% of neonatal mortality. Approximately one-third of preterm births are caused by the spontaneous onset of preterm labor. Nevertheless, 70%-90% of women diagnosed with preterm labor will not deliver within seven days. Thus, many women will be unnecessarily treated by preterm labor with risk medications. Better tools are needed to categorize women in preterm labor into high or low risk of preterm delivery. ObjectiveTo evaluate the amino-terminal pro-brain natriuretic peptide concentration in the amniotic fluid as a prognostic test to predict the risk of delivery within 48 h or 7 days and before 34+0 or 37+0 weeks in women in preterm labor. Study Design102 pregnant women presenting signs and symptoms of spontaneous preterm birth (22+0 to 34+0 weeks of gestation) were included. Amniotic fluid was obtained by amniocentesis, and amino-terminal pro-brain natriuretic peptide concentration was measured. Below normal concentration was defined as < 0.5 multiples of the median of the standard curve according to gestational age. We estimated the risk of preterm delivery risk according to normal or lower-than-normal concentrations of amino-terminal pro-brain natriuretic peptide. The predictive capacity of the test (below normal concentration of amino-terminal pro-brain natriuretic peptide) was evaluated to identify spontaneous preterm birth at 48 h or 7 days from amniocentesis, and less than 34+0 or 37+0 weeks at delivery. ResultsFor the outcome delivery within 48 h, lower than normal amino-terminal pro-brain natriuretic peptide concentration had 94.6% sensitivity, 73.8% specificity, 96% negative predictive value, and positive and negative likelihood ratios 3.6 and 0.07, respectively. For the outcome delivery within 7 days, the test had 93.9% sensitivity, 88.7% specificity, 94% negative predictive value, and positive and negative likelihood ratios 8.3 and 0.07, respectively. For the outcome of spontaneous preterm birth before 34+0 and 37+0 weeks, below normal amino-terminal pro-brain natriuretic peptide had 80% sensitivity, 83% specificity, 78% negative predictive value, positive and negative likelihood ratio 4.7 and 0.24 and 64.1% sensitivity, 91.7% specificity, 44% negative predictive value, positive and negative likelihood ratio 7.7 and 0.39 respectively. ConclusionAmong patients in spontaneous preterm labor, the detection of lower-than-normal concentrations of amino-terminal pro-brain natriuretic peptide (< 0.5 multiples of the median) in amniotic fluid has an excellent predictive capacity to identify those patients at low risk of preterm delivery within 48 h or 7 days.

Adiponectin and visfatin expression profile in extra-embryonic annexes and role during embryo development in layer and broiler chickens

Some evidence showed differences between layer and broiler embryo development. We recently showed that two adipokines, adiponectin and visfatin are expressed in the extra embryonic membranes and fluids. However, their role in the embryo development is unknown. Thus, our objectives were 1. to compare the expression of AdipoQ and its receptors AdipoR1 and AdipoR2 and visfatin in extra-embryonic annexes in broiler and layer breeders during the embryo development and 2. to investigate the role of two adipokines in embryo development in both broiler and layer breed after in ovo injection of blocking antibodies against chicken adiponectin or visfatin. We found that adiponectin, AdipoR1, AdipoR2 and visfatin were mainly more expressed in the allantoic that in amniotic membranes. In addition, these expressions increased according the stage of embryo development. We observed a higher expression in layer than in broiler of AdipoQ in allantoic membranes at ED14 and ED18, of AdipoR1 and AdipoR2 in both allantoic and amniotic membranes at ED7 and ED14 and of visfatin only in allantoic membrane from ED7 to ED18. AdipoQ and visfatin were absent in amniotic fluid at ED7 but present at ED14 or ED18 where higher concentrations were detected in layer than in broiler. Interestingly, we showed a strong positive correlation between Adipo and visfatin concentration in amniotic fluid and the body weight of embryo in both breeds. However, after in ovo injection of Adipo antibodies we did not observe any effect on the embryo mortality whereas injection of visfatin antibodies increased in a dose dependent manner the embryo mortality in both breeds. Taken together, Adipo and visfatin are higher expressed in layer than broiler in extra-embryonic membranes and amniotic fluid. Our data suggest also that visfatin could be a main regulator of embryo development.

Spatial profiling of the placental chorioamniotic membranes reveals upregulation of immune checkpoint proteins during Group B Streptococcus infection in a nonhuman primate model.

Preterm birth is a leading cause of neonatal mortality, which is often complicated by intrauterine infection and inflammation. We have established a nonhuman primate model of Group B Streptococcus (GBS, Streptococcus agalactiae) infection-associated preterm birth. Immune checkpoints are modulators of the immune response by activating or suppressing leukocyte function and are understudied in preterm birth. The objective of this study was to spatially profile changes in immune protein expression at the maternal-fetal interface during a GBS infection with a focus on immune checkpoints. Twelve nonhuman primates (pigtail macaques, Macaca nemestrina) received a choriodecidual inoculation of either: 1) 1-5 X 108 colony forming units (CFU) of hyperhemolytic/hypervirulent GBS (GBSΔcovR, N=4); 2) an isogenic/nonpigmented strain (GBS ΔcovRΔcylE, N=4); or, 3) saline (N=4). A Cesarean section was performed at preterm labor or 3 days after GBS infection or 7 days after saline inoculation. Nanostring GeoMx® Digital Spatial Profiling technology was used to segment protein expression within the amnion, chorion, and maternal decidua at the inoculation site using an immuno-oncology panel targeting 56 immunoproteins enriched in stimulatory and inhibitory immune checkpoint proteins or their protein ligands. Statistical analysis included R studio, Kruskal-Wallis, Pearson and Spearman tests. Both inhibitory and stimulatory immune checkpoint proteins were significantly upregulated within the chorioamniotic membranes and decidua (VISTA, LAG3, PD-1, CD40, GITR), as well as their ligands (PD-L1, PD-L2, CD40L; all p<0.05). Immunostaining for VISTA revealed positive (VISTA+) cells, predominantly in the chorion and decidua. There were strong correlations between VISTA and amniotic fluid concentrations of IL-1β, IL-6, IL-8, and TNF-α (all p<0.05), as well as maternal placental histopathology scores (p<0.05). Differential regulation of multiple immune checkpoint proteins in the decidua at the site of a GBS infection indicates a major perturbation in immunologic homeostasis that could benefit the host by restricting immune-driven pathologies or the pathogen by limiting immune surveillance. Protein expression of VISTA, an inhibitory immune checkpoint, was upregulated in the chorion and decidua after GBS infection. Investigating the impact of innate immune cell expression of inhibitory immune checkpoints may reveal new insights into placental host-pathogen interactions at the maternal-fetal interface.

Development of a Generic Fetal Physiologically Based Pharmacokinetic Model and Prediction of Human Maternal and Fetal Organ Concentrations of Cefuroxime.

Physiologically based pharmacokinetic (PBPK) models for pregnant women have recently been successfully used to predict maternal and umbilical cord pharmacokinetics (PK). Because there is very limited opportunity for conducting clinical and PK investigations for fetal drug exposure, PBPK models may provide further insights. The objectives of this study were to extend a whole-body pregnancy PBPK model by multiple compartments representing fetal organs, and to predict the PK of cefuroxime in the maternal and fetal plasma, the amniotic fluid, and several fetal organs. To this end, a previously developed pregnancy PBPK model for cefuroxime was updated using the open-source software Open Systems Pharmacology (PK-Sim®/MoBi®). Multiple compartments were implemented to represent fetal organs including brain, heart, liver, lungs, kidneys, the gastrointestinal tract (GI), muscles, and fat tissue, as well as another compartment lumping organs and tissues not explicitly represented. This novel PBPK model successfully predicted cefuroxime concentrations in maternal blood, umbilical cord, amniotic fluid, and several fetal organs including heart, liver, and lungs. Further model validation with additional clinical PK data is needed to build confidence in the model. Being developed with an open-source software, the presented generic model can be freely re-used and tailored to address specific questions at hand, e.g., to assist the design of clinical studies in the context of drug research or to predict fetal organ concentrations of chemicals in the context of fetal health risk assessment.

Comparative analysis of the use of selective serotonin reuptake inhibitors during pregnancy and the postpartum period. Safety of sertraline

Depressive disorders and anxiety are the most common mental disorders in the perinatal period, occurring in 14–23% of women. Antidepressants from the selective serotonin reuptake inhibitor (SSRI) group are considered the drugs of choice for the treatment of these disorders. Although SSRIs are the best-studied antidepressants used in pregnant women, there are still conflicting opinions about their effect on pregnancy course and outcomes of pregnancy. At the same time, the risks associated with their use in pregnant women are often exaggerated, while the risks of untreated mental disorders are underestimated. SSRI use during pregnancy has been associated with a modestly increased risk of adverse events, including postpartum haemorrhage, miscarriage, preterm birth, cesarean delivery, fetuses small for their gestational age and low birth weight, low Apgar scores, and increased rates of neonatal hospitalization to intensive care units. However, depressive and anxiety disorders in the mothers themselves also contribute to similar outcomes, so it is not always possible to distinguish the contribution of medication and illness. Comparative data on the effects of different SSRI medications on outcome for the mother and fetus during pregnancy are limited, but paroxetine and fluoxetine are reported to pose the greatest risk to the fetus/neonate. Information on the safety of citalopram and escitalopram during pregnancy and lactation is limited. Citalopram has the highest concentrations in amniotic fluid of all SSRIs. According to current data, sertraline has the most favourable safety profile during pregnancy and lactation, which is explained by its low penetration through the placenta. Sertraline concentrations in the fetus are approximately 1/3 of those in maternal plasma. Sertraline has the lowest concentrations in breast milk of all SSRIs and is associated with a low risk of adverse effects in a baby, making it the drug of choice for nursing mothers. In conclusion, sertraline is one of the best studied SSRIs and has a favourable safety profile for both the mother and the fetus/neonate.

Association of chromosomal abnormalities with prenatal exposure to heavy metals: A nested case-control study in high-risk pregnant women in China

Prenatal exposure to heavy metals causes multiple hazards to fetal growth and development. Epidemiological studies on the association between heavy metals and fetal chromosomal abnormalities (CAs) are lacking. We conducted a nested case-control study in a cohort of high-risk pregnant women in China from September 2018 to June 2021. A total of 387 participants were diagnosed with fetal CAs in the case group and 699 were diagnosed with a normal karyotype in the control group. Amniotic fluid concentrations of 10 metals (barium, cobalt, antimony, manganese, ferrum, copper, selenium, strontium, vanadium, and chromium) were measured using inductively coupled plasma-mass spectrometry. We applied quantile g-computation and weighted quantile sum regression to assess the overall effect of metal mixtures and identify metals with significant weight. Logistic and Poisson regression analyses were used to estimate the effects of metals on CAs and CAs subtypes. Our results showed that the metal mixture concentrations were positively associated with the risk of fetal CAs. In adjusted logistic models, Sb was associated with fetal CAs (OR=1.15, 95% CI: 1.02–1.30), and revealed a linear dose-response relationship between Sb level and the risk of fetal CAs. Additionally, the exploratory analysis revealed that Sb levels were associated with Klinefelter syndrome (OR=1.452, 95% CI: 1.063–1.984) and Turner syndrome (OR=1.698; 95% CI,1.048–2.751). Our study revealed that metal mixtures are associated with a higher risk of fetal CAs and that this association may be driven primarily by Sb. Moreover, we provide a genetic perspective on the effects of heavy metals on sexual development in humans.

Amniotic fluid rubidium concentration association with newborn birthweight: a maternal-neonatal pilot study

Although most biological systems, including human tissues, contain rubidium, its biogeochemical functions and possible role in neonatal birthweight are largely unknown. An animal study indicated a correlation between rubidium deficiency in the maternal diet and lower newborn birthweight. This pilot study measured rubidium concentrations in amniotic fluid during the second trimester of (low-risk) pregnancy and investigated potential correlations between rubidium levels and third-trimester newborn birthweight-small for gestational age, appropriate for gestational age, and large for gestational age-and between preterm birth and term birth in uncomplicated pregnancies. This prospective, single-center study investigated a possible relationship between rubidium concentration in second-trimester amniotic fluid and third-trimester birthweight percentile. Amniotic fluid (at a median gestational age of 19 weeks) was sampled to determine rubidium concentration. Maternal and newborn characteristics were obtained from participant and delivery records. After screening 173 pregnant women, 99 amniotic fluid samples were evaluated. Midpregnancy median rubidium concentrations were significantly lower among newborns that were classified as small for gestational age than among newborns that were classified as appropriate for gestational age (106 vs 136 μg/L; P<.01). Based on a logistic regression random forest model, amniotic fluid rubidium was identified as a significant contributing factor to appropriate-for-gestational-age birthweight with 54% of the total contribution. Amniotic fluid rubidium concentration seems to be a strong predictor of appropriate-for-gestational-age birthweight and a potential marker for newborn birthweight classifications. In particular, low rubidium concentrations in amniotic fluid during midpregnancy are linked to third-trimester lower birthweight percentile. These findings could potentially serve as a valuable tool for early identification of pregnancy outcomes. Further investigation is necessary to fully explore the effect of rubidium on fetal development.

Cotinine concentrations in maternal serum and amniotic fluid during pregnancy and risk of testicular germ cell cancer in the offspring: A prospective nested case-control study.

Maternal smoking in pregnancy may increase the risk of testicular germ cell cancer (TGCC) in offspring, but current evidence remains inconclusive. We performed a nested case-control study using cotinine measurements in maternal serum and amniotic fluid as a biomarker for tobacco exposure during pregnancy. A total of 654 males with maternal serum (n = 359, ncases/controls = 71/288) and/or amniotic fluid (n = 295, ncases/controls = 66/229) samples were included. Data on TGCC diagnoses and relevant covariates were derived from nationwide Danish health registries. Cotinine was quantified by liquid chromatography tandem mass spectrometry. An adapted cox regression model estimated the risk of TGCC considering active and inactive tobacco use defined according to cotinine concentrations of <, ≥15 ng/ml. Overall, the concentrations of cotinine were comparable in maternal serum and amniotic fluid (medianserum/amniotic fluid : 2.1/2.6 ng/ml). A strong statistically significant correlation was detected in 14 paired samples (Spearman rho: 0.85). Based on maternal serum cotinine concentrations, exposure to active tobacco use was not associated with risk of TGCC in offspring (HR 0.88, 95% CI 0.51; 1.52). Similarly, based on amniotic fluid cotinine concentrations, exposure to active tobacco use was not associated with risk of TGCC (HR 1.11, 95% CI 0.64; 1.95). However, different risks were observed for seminomas and nonseminomas in both matrices, but none were statistically significant. Our findings did not provide convincing evidence supporting that exposure to tobacco during pregnancy is associated with TGCC.

Quantification of sertraline maternal/fetal ratio and amniotic fluid concentration using a pregnancy physiologically based pharmacokinetic model.

Selective serotonin reuptake inhibitors (SSRIs) are indicated for a variety of psychiatric conditions which may require treatment during pregnancy. Knowledge of appropriate SSRI dosages that maintain maternal therapeutic benefit and minimize fetal risk are needed. Assessment of fetal exposure to drugs is challenging since sampling is often limited to a single concentration from the umbilical cord at delivery. Physiologically based pharmacokinetic (PBPK) modelling provides a non-invasive approach to quantify exposure in pregnancy. We incorporated sertraline clearances mediated by passive diffusion, placental efflux transporters P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP) into our previously published pregnancy PBPK model for sertraline. Simulations were performed for various sertraline doses (25-200 mg) at 40 weeks gestational age to predict the minimum (Cmin ), maximum (Cmax ) and average (Cavg ) sertraline maternal and fetal plasma concentrations and evaluated them against observed maternal and cord concentrations obtained at delivery from five clinical studies. The accuracy of the PBPK predictions as indicated by the average fold error (AFE) value for Cmax , Cmin and Cavg for maternal plasma sertraline concentrations at delivery was 1.7, 1.2 and 1.4, respectively. The AFE for the Cmax , Cmin and Cavg for cord blood sertraline concentration at delivery was 1.2, 1 and 1.1, respectively. The AFE for cord-maternal sertraline concentration ratio at delivery for Cmax , Cmin and Cavg was 0.7, 0.9 and 0.8, respectively. The PBPK model we developed may serve as a guide for maternal sertraline dose adjustment during pregnancy considering changes in exposures for both mother and fetus.

Amniotic fluid metabolomics identifies impairment of glycerophospholipid and amino acid metabolism during congenital Zika syndrome development.

Our main goal is to identify the alterations in the amniotic fluid (AF) metabolome in Zika virus (ZIKV)-infected patients and their relation to congenital Zika syndrome (CZS) progression. We applied an untargeted metabolomics strategy to analyze seven AF of pregnant women: healthy women and ZIKV-infected women bearing non-microcephalic and microcephalic fetuses. Infected patients were characterized by glycerophospholipid metabolism impairment, which is accentuated in microcephalic phenotypes. Glycerophospholipid decreased concentration in AF can be a consequence of intracellular transport of lipids to the placental or fetal tissues under development. The increased intracellular concentration of lipids can lead to mitochondrial dysfunction and neurodegeneration caused by lipid droplet accumulation. Furthermore, the dysregulation of amino acid metabolism was a molecular fingerprint of microcephalic phenotypes, specifically serine, and proline metabolisms. Both amino acid deficiencies were related to neurodegenerative disorders, intrauterine growth retardation, and placental abnormalities. This study enhances our understanding of the development of CZS pathology and sheds light on dysregulated pathways that could be relevant for future studies.

Effects of firocoxib, trimethoprim sulfamethoxazole and altrenogest on inflammation and foal survival after administration to mares with experimentally induced placentitis

Bacterial placentitis is a leading cause of pregnancy loss in mares. Infection, inflammation and prostaglandin production induce preterm delivery leading to poor foal survival. Treatment strategies for placentitis that control infection and inhibit inflammation are vital. The goals of this study were: 1. Assess the anti-inflammatory effects of firocoxib, trimethoprim sulfamethoxazole (TMS) and altrenogest administered to mares with placentitis; 2. Determine the efficacy of firocoxib, TMS and altrenogest for improving foal outcome in mares with placentitis. We hypothesized that this treatment would significantly decrease pro-inflammatory mediators in fetal fluids and tissues when compared to untreated, infected mares and would also improve foal survival. Placentitis was induced in 13 pregnant mares (270-300 days gestation; via intracervical inoculum of Streptococcus zooepidemicus). Mares were randomly assigned to: infected/untreated (UNTREATED; n=6); or infected/treated groups (TREATED; n=7; administered firocoxib, TMS, and altrenogest from the onset of clinical signs of placentitis until abortion/delivery). Fetal fluids, membranes and tissue samples were obtained at abortion/delivery for quantification of TNF-α, IL-1β, IL-10, IL-6, IL-8, PGE2, PGF2α, and matrix metalloproteinases (MMP) -1, -3, -9 using ELISA (fluids) and real-time PCR (tissues). P < 0.05 was considered significant. Gestation length and foal survival were significantly higher in TREATED (mean days post-infection = 40; 7/7 live foals) than UNTREATED (mean days post-infection = 14; 2/6 live foals). Administration of firocoxib, TMS andaltrenogest resulted in significantly lower amniotic fluid concentrations of IL-1β, IL-6, PGE2 and PGF2α in TREATED than UNTREATED. Amniotic fluid IL-10 concentration tended (P < 0.1) to be lower in TREATED than UNTREATED. Cytokine and prostaglandin concentrations in allantoic fluid were not different between groups except PGE2 which tended (P < 0.1) to be lower in TREATED than UNTREATED. The relative quantity of cytokine and MMP mRNA expression in fetal membranes and foal tissues (non-survivors) was log-transformed and estimated marginal means determined. Fetal membranes from TREATED had significantly lower mRNA expression for IL-8 and TNFα compared to membranes from UNTREATED. mRNA expression of IL-1β, IL-6 and MMP3 tended (P < 0.1) to be lower in membranes from TREATED than UNTREATED. mRNA expression of IL-1β, IL-8, IL-10, TNFα, MMP1, 3 and 9 were significantly different between fetal membrane sites. In non-surviving foals, IL-8 mRNA expression was significantly lower in liver than lung while MMP3 expression was significantly higher in liver than lung or spleen. These data provide physiologic evidence that administration of firocoxib, TMS and altrenogest to mares with placentitis has broad anti-inflammatory effects and may improve foal survival. Research funding: Grayson Jockey Club Research Foundation; Merial Animal Health; Kentucky Thoroughbred Breeders Association, University of Florida, University of Georgia.

Amniotic fluid CD36 in pregnancies complicated by spontaneous preterm delivery: a retrospective cohort study

Objective The aim of this study was to evaluate CD36 concentrations in amniotic fluid in pregnancies complicated by spontaneous delivery with intact fetal membranes (preterm labor, PTL) and preterm prelabor rupture of membranes (PPROM) with respect to the presence of the intra-amniotic infection. Methods A total of 80 women with PPROM and 71 with PTL were included in the study. Amniotic fluid samples were obtained by transabdominal amniocentesis. Amniotic fluid CD36 concentrations were assessed by enzyme-linked immunosorbent assay. Microbial colonization of the amniotic cavity (MIAC) was determined by the cultivation and non-cultivation approach. Intra-amniotic inflammation (IAI) was defined as an amniotic fluid bedside interleukin-6 concentration ≥3000 pg/mL. Intra-amniotic infection was characterized by the presence of both MIAC and IAI. Results Women with PPROM with intra-amniotic infection had higher amniotic fluid CD36 concentrations than women without infection (with infection: median 346 pg/mL, IQR 262–384 vs. without infection: median 242 pg/mL, IQR 199–304; p = .006) A positive correlation between amniotic fluid CD36 concentrations and interleukin-6 concentrations was found (rho = 0.48; p < .0001). In PTL pregnancies, no statistically significant difference was found in the amniotic fluid level of CD36 between intra-amniotic infection, sterile IAI, and negative amniotic fluid. Conclusions The presence of intra-amniotic infection is characterized by higher amniotic fluid CD36 concentrations in pregnancies complicated by PPROM. An amniotic fluid CD36 cutoff value of 252.5 pg/mL was found to be optimal for the prediction of intra-amniotic infection. In PTL pregnancies, no statistically significant change in CD36 concentration was found with respect to the presence of intra-amniotic infection.

Is There a Correlation between Apelin and Insulin Concentrations in Early Second Trimester Amniotic Fluid with Fetal Growth Disorders?

Fetal growth disturbances place fetuses at increased risk for perinatal morbidity and mortality. As yet, little is known about the basic pathogenetic mechanisms underlying deranged fetal growth. Apelin is an adipokine with several biological activities. Over the past decade, it has been investigated for its possible role in fetal growth restriction. Most studies have examined apelin concentrations in maternal serum and amniotic fluid in the third trimester or during neonatal life. In this study, apelin concentrations were examined for the first time in early second-trimester fetuses. Another major regulator of tissue growth and metabolism is insulin. This was a prospective observational cohort study. We measured apelin and insulin concentrations in the amniotic fluid of 80 pregnant women who underwent amniocentesis in the early second trimester. Amniotic fluid samples were stored in appropriate conditions until delivery. The study groups were then defined, i.e., gestations with different fetal growth patterns (SGA, AGA, and LGA). Measurements were made using ELISA kits. Apelin and insulin levels were measured in all 80 samples. The analysis revealed statistically significant differences in apelin concentrations among groups (p = 0.007). Apelin concentrations in large for gestational age (LGA) fetuses were significantly lower compared to those in AGA and SGA fetuses. Insulin concentrations did not differ significantly among groups. A clear trend towards decreasing apelin concentrations as birthweight progressively increased was identified. Amniotic fluid apelin concentrations in the early second trimester may be useful as a predictive factor for determining the risk of a fetus being born LGA. Future studies are expected/needed to corroborate the present findings and should ideally focus on the potential interplay of apelin with other known intrauterine metabolic factors.

Transplacental Transport of Artificial Sweeteners

The prevalence of obesity is increasing, and the origins of obesity and metabolic dysfunction may be traced back to fetal life. Currently, overweight pregnant women are advised to substitute sugar-sweetened beverages with diet drinks containing artificial sweeteners. Recent evidence suggests that the consumption of artificial sweeteners during pregnancy increases the risk of obesity in the child, but the mechanism is unknown. We hypothesized the transportation of artificial sweeteners across the placenta into the fetal circulation and the amniotic fluid. We included 19 pregnant women who were given an oral dose of acesulfame, cyclamate, saccharin, and sucralose immediately before a planned caesarean section. Nine women were included as controls, and they refrained from an intake of artificial sweeteners. The maternal and fetal blood and amniotic fluid were collected during the caesarean section, and concentrations of artificial sweeteners were measured using mass spectrometry. We found a linear relationship between the fetal plasma concentrations of artificial sweeteners and the maternal plasma concentrations, with adjusted coefficients of 0.49 (95% CI: 0.28–0.70) for acesulfame, 0.72 (95% CI: 0.48–0.95) for cyclamate, 0.51 (95% CI: 0.38–0.67) for saccharin, and 0.44 (95% CI: 0.33–0.55) for sucralose. We found no linear relationship between amniotic fluid and fetal plasma concentrations, but there were positive ratios for all four sweeteners. In conclusion, the four sweeteners investigated all crossed the placenta and were present in the fetal circulation and amniotic fluid.

Evidence for the participation of CHCHD2/MNRR1, a mitochondrial protein, in spontaneous labor at term and in preterm labor with intra-amniotic infection

Objective Intra-amniotic inflammation (IAI), associated with either microbe (infection) or danger signals (sterile), plays a major role in the pathophysiology of preterm labor and delivery. Coiled-Coil-Helix-Coiled-Coil-Helix Domain Containing 2 (CHCHD2) [also known as Mitochondrial Nuclear Retrograde Regulator 1 (MNRR1)], a mitochondrial protein involved in oxidative phosphorylation and cell survival, is capable of sensing tissue hypoxia and inflammatory signaling. The ability to maintain an appropriate energy balance at the cellular level while adapting to environmental stress is essential for the survival of an organism. Mitochondrial dysfunction has been observed in acute systemic inflammatory conditions, such as sepsis, and is proposed to be involved in sepsis-induced multi-organ failure. The purpose of this study was to determine the amniotic fluid concentrations of CHCHD2/MNRR1 in pregnant women, women at term in labor, and those in preterm labor (PTL) with and without IAI. Methods This cross-sectional study comprised patients allocated to the following groups: (1) mid-trimester (n = 16); (2) term in labor (n = 37); (3) term not in labor (n = 22); (4) PTL without IAI who delivered at term (n = 25); (5) PTL without IAI who delivered preterm (n = 47); and (6) PTL with IAI who delivered preterm (n = 53). Diagnosis of IAI (amniotic fluid interleukin-6 concentration ≥2.6 ng/mL) included cases associated with microbial invasion of the amniotic cavity and those of sterile nature (absence of detectable bacteria, using culture and molecular microbiology techniques). Amniotic fluid and maternal plasma CHCHD2/MNRR1 concentrations were determined with a validated and sensitive immunoassay. Results (1) CHCHD2/MNRR1 was detectable in all amniotic fluid samples and women at term without labor had a higher amniotic fluid CHCHD2/MNRR1 concentration than those in the mid-trimester (p = 0.003); (2) the amniotic fluid concentration of CHCHD2/MNRR1 in women at term in labor was higher than that in women at term without labor (p = 0.01); (3) women with PTL and IAI had a higher amniotic fluid CHCHD2/MNRR1 concentration than those without IAI, either with preterm (p < 0.001) or term delivery (p = 0.01); (4) women with microbial-associated IAI had a higher amniotic fluid CHCHD2/MNRR1 concentration than those with sterile IAI (p < 0.001); (5) among women with PTL and IAI, the amniotic fluid concentration of CHCHD2/MNRR1 correlated with that of interleukin-6 (Spearman’s Rho = 0.7; p < 0.001); and (6) no correlation was observed between amniotic fluid and maternal plasma CHCHD2/MNRR1 concentrations among women with PTL. Conclusion CHCHD2/MNRR1 is a physiological constituent of human amniotic fluid in normal pregnancy, and the amniotic concentration of this mitochondrial protein increases during pregnancy, labor at term, and preterm labor with intra-amniotic infection. Hence, CHCHD2/MNRR1 may be released into the amniotic cavity by dysfunctional mitochondria during microbial-associated IAI.

Maternal-fetal biodisposition of buprenorphine and its metabolites in opioid dependent pregnant individuals

To determine the concentration of buprenorphine (BUP) and its metabolites in maternal and fetal samples and investigate the relationship between BUP concentrations and neonatal opioid withdrawal syndrome (NOWS). Matched maternal and fetal samples were collected from 12 individuals maintained on BUP for opioid use disorder during pregnancy. All delivered at term (≥37 weeks gestation) via cesarean section. Samples were collected at delivery and included maternal plasma (PL), amniotic fluid (AF), and umbilical cord blood (CB). Concentrations of BUP, its biologically active metabolite norbuprenorphine (NBUP) and its inactive metabolites glucuronide forms of buprenorphine (BUP-Gluc) and norbuprenorphine (NBUP-Gluc) were determined by liquid chromatography-mass spectrometry. The concentrations of BUP and its metabolites were compared between neonates who developed NOWS, defined as withdrawal symptoms requiring pharmacologic treatment, and those who did not. Five (42%) of neonates born to mothers in the study developed NOWS. There was no difference in gestational age, birthweight or other baseline characteristics between the two groups. All individuals in the study were receiving 16 mg daily of BUP at delivery. BUP concentrations were higher in cord blood (0.65 v 0.31ng/mL, p=0.01) and amniotic fluid (0.64 v 0.0 ng/mL, p=0.04) in neonates who developed NOWS, compared to those who did not. The concentrations of NBUP were also higher in neonates who developed NOWS (2.01 v 0.0ng/mL, p=0.03). There were no differences in maternal plasma concentrations between the two groups. Despite similar maternal dosing and plasma concentrations, NBUP concentrations in AF and BUP concentrations in AF and CB were higher for infants later diagnosed with NOWS, suggesting accumulation of the drug in the fetal compartment. Further studies are needed to determine the mechanisms and whether analysis of these specimens at delivery may help predict NOWS in buprenorphine-exposed neonates.

Comparative Study on Umbilical Cord Serum Prolactin Levels of Normal Preterm Neonates and Preterm Neonates with Respiratory Distress Syndrome

Introduction: Respiratory distress syndrome (RDS) is the most common cause of respiratory distress, morbidity, and mortality associated with premature delivery. Prolactin is present in very high concentrations in amniotic fluid, and prolactin receptors are present in lung preparations. Prolactin acts as a direct trigger of lecithin synthesis or in a “chain reaction” along the hypothalamic–pituitary–adrenal axis. Thus, prolactin seems to have a role in lung maturation along with many other factors through a complex mechanism. Aim: The aim of this study was to evaluate and compare the umbilical cord serum prolactin levels of preterm infants with and without RDS. Materials and Methods: This was a hospital-based comparative observational study carried out at neonatal units of the department of pediatric medicine in a tertiary care center from April 2019 to May 2020. Out of a total of 80 preterm infants, 40 developed RDS and other 40 served as controls. Umbilical cord blood prolactin levels in neonates were measured in both groups and compared. Chi-square test and an unpaired Student’s t-test were used for statistical analysis. Probability was considered significant if &lt;0.05. Results: The mean weight of babies was 1312.50 ± 134.30 g in the normal preterm group and 1285.80 ± 314.76 g in the preterm RDS group (P = 0.622). The mean gestational age was 32.1 ± 1.39 weeks in the normal preterm group and 31.77 ± 2.10 weeks in the preterm RDS group (P = 0.418). The mean serum prolactin levels were slightly high in the female preterm RDS group (P = 0.533). The mean cord prolactin levels in all the gestational ages were higher in normal preterm as compared to preterm with RDS (P &lt; 0.05). Conclusions: Prolactin level was significantly associated with gestational age and birth weight. Low prolactin levels were significantly associated with the development of RDS in preterm babies. The easily available cord blood can be used to measure prolactin levels at birth to predict RDS.

Short-term toxicity studies of thallium (I) sulfate administered in drinking water to Sprague Dawley rats and B6C3F1/N mice.

Thallium is a heavy metal that is known to induce a broad spectrum of adverse health effects in humans including alopecia, neurotoxicity, and mortality following high dose acute poisoning events. Widespread human exposure to thallium may occur via consumption of contaminated drinking water; limited toxicity data are available to evaluate the corresponding public health risk. To address this data gap, the Division of Translational Toxicology conducted short-term toxicity studies of a monovalent thallium salt, thallium (I) sulfate. Thallium (I) sulfate was administered via dosed drinking water to time-mated Sprague Dawley (Hsd:Sprague Dawley® SD®) rats (F0 dams) and their offspring (F1) from gestation day (GD) 6 until up to postnatal day (PND) 28 at concentrations of 0, 3.13, 6.25, 12.5, 25, or 50mg/L, and adult male and female B6C3F1/N mice for up to 2 weeks at concentrations of 0, 6.25, 12.5, 25, 50, or 100mg/L. Rat dams in the 50mg/L exposure group were removed during gestation, and dams and offspring in the 25mg/L exposure group were removed on or before PND 0 due to overt toxicity. Exposure to thallium (I) sulfate at concentrations ≤12.5mg/L did not impact F0 dam body weights, maintenance of pregnancy, littering parameters, or F1 survival (PND 4-28). However, in F1 pups, exposure to 12.5mg/L thallium (I) sulfate resulted in decreased body weight gains relative to control rats and onset of whole-body alopecia. Measurement of thallium concentrations in dam plasma, amniotic fluid, fetuses (GD 18), and pup plasma (PND 4) indicated marked maternal transfer of thallium to offspring during gestation and lactation. Mice exposed to 100mg/L thallium (I) sulfate were removed early due to overt toxicity, and mice exposed to ≥25mg/L exhibited exposure concentration-related decreases in body weight. Lowest-observed-effect levels of 12.5mg/L (rats) and 25mg/L (mice) were determined based on the increased incidence of clinical signs of alopecia in F1 rat pups and significantly decreased body weights for both rats and mice.

Amniotic fluid soluble CD93 is elevated in the presence of intra-amniotic inflammation in preterm prelabor rupture of the fetal membranes.

To determine the soluble form of CD93 (sCD93) concentration in amniotic fluid from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on the presence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation. A total of 144 women with a singleton pregnancy complicated by PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. MIAC was determined by the combination of cultivation and non-cultivation techniques. Intra-amniotic inflammation was characterized as a concentration of interleukin-6 3,000 pg/mL in amniotic fluid. Women were categorized in the following groups: i) intra-amniotic infection (both MIAC and intra-amniotic inflammation), ii) sterile intra-amniotic inflammation (intra-amniotic inflammation per se), iii) colonization of the amniotic cavity (MIAC per se), and iv) negative amniotic fluid (without both MIAC and intra-amniotic inflammation). Levels of sCD93 in amniotic fluid were assessed by ELISA. A difference in the levels of sCD93 in amniotic fluid was found among the groups of women with intra-amniotic infection, sterile intra-amniotic inflammation, colonization of the amniotic cavity, and negative amniotic fluid (intra-amniotic infection: median 22.3 ng/mL, sterile intra-amniotic inflammation: median 21.0 ng/mL, colonization of the amniotic cavity: 8.7 ng/mL, negative: median 8.7 ng/mL; P < 0.0001). Intra-amniotic inflammation in PPROM, irrespectively of the presence or absence of MIAC, is associated with the elevation of the level of sCD93 in amniotic fluid.