Neonatal Cord Research Articles

DNA methylation and expression profiles of placenta and umbilical cord blood reveal the characteristics of gestational diabetes mellitus patients and offspring

BackgroundGestational diabetes mellitus (GDM) is a common pregnancy-specific disease and is growing at an alarming rate worldwide, which can negatively affect the health of pregnant women and fetuses. However, most studies are limited to one tissue, placenta or umbilical cord blood, usually with one omics assay. It is thus difficult to systematically reveal the molecular mechanism of GDM and the key influencing factors on pregnant women and offspring.ResultsWe recruited a group of 21 pregnant women with GDM and 20 controls without GDM. For each pregnant woman, reduced representation bisulfite sequencing and RNA-seq were performed using the placenta and paired neonatal umbilical cord blood specimens. Differentially methylated regions (DMRs) and differentially expressed genes (DEGs) were identified with body mass index as a covariate. Through the comparison of GDM and control samples, 2779 and 141 DMRs, 1442 and 488 DEGs were identified from placenta and umbilical cord blood, respectively. Functional enrichment analysis showed that the placenta methylation and expression profiles of GDM women mirrored the molecular characteristics of “type II diabetes” and “insulin resistance.” Methylation-altered genes in umbilical cord blood were associated with pathways “type II diabetes” and “cholesterol metabolism.” Remarkably, both DMRs and DEGs illustrated significant overlaps among placenta and umbilical cord blood samples. The overlapping DMRs were associated with “cholesterol metabolism.” The top-ranking pathways enriched in the shared DEGs include “growth hormone synthesis, secretion and action” and “type II diabetes mellitus.”ConclusionsOur research demonstrated the epigenetic and transcriptomic alternations of GDM women and offspring. Our findings emphasized the importance of epigenetic modifications in the communication between pregnant women with GDM and offspring, and provided a reference for the prevention, control, treatment, and intervention of perinatal deleterious events of GDM and neonatal complications.

The relationship between clinically significant neonatal hypoglycemia and cord blood c-peptide levels in neonates of mothers with type 1 diabetes

The relationship between clinically significant neonatal hypoglycemia and cord blood c-peptide levels in neonates of mothers with type 1 diabetes

Neurotransmitter phenotype switching by spinal excitatory interneurons regulates locomotor recovery after spinal cord injury.

Severe spinal cord injury in adults leads to irreversible paralysis below the lesion. However, adult rodents that received a complete thoracic lesion just after birth demonstrate proficient hindlimb locomotion without input from the brain. How the spinal cord achieves such striking plasticity remains unknown. In this study, we found that adult spinal cord injury prompts neurotransmitter switching of spatially defined excitatory interneurons to an inhibitory phenotype, promoting inhibition at synapses contacting motor neurons. In contrast, neonatal spinal cord injury maintains the excitatory phenotype of glutamatergic interneurons and causes synaptic sprouting to facilitate excitation. Furthermore, genetic manipulation to mimic the inhibitory phenotype observed in excitatory interneurons after adult spinal cord injury abrogates autonomous locomotor functionality in neonatally injured mice. In comparison, attenuating this inhibitory phenotype improves locomotor capacity after adult injury. Together, these data demonstrate that neurotransmitter phenotype of defined excitatory interneurons steers locomotor recovery after spinal cord injury.

Role of the Dopamine Subtype 1 Receptor (D1R) Modulation of the Ih Current in Rhythmic Spinal Mammalian Motor Networks

Restless Legs Syndrome (RLS) is a chronic sensorimotor disorder that has been recently associated with decreased expression of inhibitory Gi‐coupled adenosine A1 receptors (A1Rs), leading to increased dopamine release (hyperdopaminergic signaling) within the central nervous system. Previous studies have shown that A1Rs specifically inhibit the Gs‐coupled dopamine D1 receptor (D1R) subtype through a functional A1R‐D1R heteromer receptor complex. Furthermore, D1Rs control spinal motor neuron (MN) excitability by increasing cyclic adenosine monophosphate (cAMP) concentration and opening hyperpolarization‐activated cyclic nucleotide‐gated non‐selective cation channel (HCN). This finding raises the possibility of increased HCN activity underlying some of the motor dysfunctions associated with periodic leg movements (PLMS) during sleep in RLS. Thus, this study aims to elucidate the importance of the D1 receptor subtype on modulating the hyperpolarization‐activated cation current (Ih) in spinal mammalian motor networks. I hypothesized that blocking Ih in the presence of a D1R agonist (enhanced dopaminergic modulation) would slow locomotor activity and disrupt MN bursting stability. Neonatal (P0‐P5; n = 7) mouse spinal cord preparations were used to measure MN bursting activity patterns including amplitude, duration, and cycle period of extracellular ventral root (L2 and L5) recordings via suction electrodes. Treatment conditions include serotonin (9‐15 μM 5‐HT) and N‐methyl‐D‐aspartate (6 μM NMDA) with and without a D1R agonist (10 μM SKF38393) to assess the effect of D1R on rhythmic bursting. Subsequently, an Ih blocker (1 μM ZD7288) was added to observe the importance of Ih in modulating MN burst activity. For both L2 and L5 roots, blocking the Ih current led to increases in burst amplitude, duration, and cycle period in respect to the control 5‐HT/NMDA condition. Our results propose that D1Rs play a significant role in modulating Ih by increasing cAMP levels and opening HCN1 channels at a more depolarized membrane potential. Future experiments involve: 1) using TM5 transmembrane disrupting peptide to disrupt A1R‐D1R heteromers in spinal MNs (and use TM7 as a negative control) to confirm if dopamine modulation of Ih activity is linked to the A1R‐D1R heteromer complexes and 2) applying adenyl cyclase activators and blockers to verify that the modulation of the Ih current is through the D1R‐mediated Gs second messenger pathway.

The effects of temperature changes on the mammalian spinal locomotor CPG network and the role of TRP channels in functional compensation

Preliminary data suggests that spinal cord neural networks that contain intact sensory pathways, which include dorsal root ganglia (DRG) and thermal transient receptor potential (TRP) channels are more resilient to gradual temperature changes than isolated preparations. Moreover, how acute temperature changes affect mammalian CPG‐driven motor activity remains uncertain. We studied how acute and gradual temperature changes impact the stability of motor networks with or without sensory pathways. More specifically, we aim to investigate: 1. the difference between how acute temperature changes versus gradual temperature changes can have an impact on the stability and robustness of motor networks; 2. how sensory inputs and TRP channels affect the adaptation of spinal central pattern generator (CPG) networks to changes in temperature. We compared changes in motor neuron bursting activity between ventral L2 and L5 roots of neonatal mouse spinal cord preparations under acute (from 20℃ directly to 29℃) and gradual (gradually from 20℃, to 23℃, to 26℃, and to 29℃) temperature change conditions. Spinal cord preparations both with and without DRGs were compared. We found that the locomotor activities were increased as temperature increase under both chronic and acute changes; however, spinal cord preparations without intact DRG showed activities with less stable phasing and more overall motor bursting variability compared to those preparations with intact DRGs (on L3, L4, and L5 spinal levels). Our results suggest that the lumbar spinal locomotor CPG network activity shows more susceptibility and less adaptation to temperature changes in the absence of sensory feedback provided by DRG sensory afferents. Further studies will be conducted in the presence of TRP channel blockers to preparations with DRGs to confirm the potential compensatory role that sensory feedback afferents convey to spinal CPG networks.

Assessment of Neonatal Cord Blood SARS-CoV-2 Antibodies after COVID-19 Vaccination in Pregnancy: A Prospective Cohort Study.

Introduction Maternally derived antibodies are a key element of neonatal immunity. So far, limited data has shown transplacental transmission of antibodies after coronavirus disease 2019 (COVID-19) vaccination with BNT162b2 in the third trimester. Our aim was to detect vertically transferred immunity after COVID-19 vaccination with BNT162b2 (Comirnaty, BioNTech-Pfizer) or mRNA-1273 (Spikevax, Moderna) in the first, second or third trimester of pregnancy, and investigate the impact of maternal characteristics on umbilical cord antibody titre in newborns after delivery. Study Design Women who gave birth in our department and were vaccinated against COVID-19 during pregnancy were enrolled in CRONOS Satellite, a subproject of the German COVID-19-Related Obstetric and Neonatal Outcome Study. The titre of immunoglobulin G (IgG) antibodies to the receptor-binding domain of the SARS-CoV-2 spike protein was quantified in umbilical cord blood using the SARS-CoV-2 IgG II Quant immunoassay. Correlations between antibody titre and variables, including week of pregnancy when vaccinated, interval between vaccination and delivery, age and body mass index (BMI) were assessed with Spearmanʼs rank correlation. A follow-up was conducted by phone interview 4 – 6 weeks after delivery. Results The study cohort consisted of 70 women and their 74 newborns. Vaccine-generated antibodies were present in all samples, irrespective of the vaccination type or time of vaccination. None of the parameters of interest showed a meaningful correlation with cord blood antibody concentrations (rho values < 0.5). No adverse outcomes (including foetal malformation) were reported, even after vaccination in the first trimester. Conclusions Transplacental passage of SARS-CoV-2 antibodies from mother to child was demonstrated in all cases in the present study. It can therefore be assumed that the newborns of mothers vaccinated at any time during pregnancy receive antibodies via the placenta which potentially provide them with protection against COVID-19. This is an additional argument when counselling pregnant women about vaccination in pregnancy.

Effect of maternal anaemia on status of iron stores in neonates and anthropometry

Objective: To evaluate the effect of maternal IDA on neonatal hemoglobin and iron stores and also to assess its correlation with neonatal anthropometric characteristics. Study Design: This was a cross-sectional study conducted at a tertiary care hospital for a period of 18 months. The study enrolled total 112 pregnant women who were diagnosed with IDA, and also included babies of enrolled women for correlation of neonatal iron status and anthropometric variables. Maternal prenatal blood samples, during the first stage of labour, and neonatal cord blood samples were collected for hemoglobin and serum ferritin analysis. Results: Mean age of enrolled pregnant women was 25.81 ± 4.67 years. Among all, 65.17% women had moderate to severe anemia. The mean hemoglobin and serum ferritin levels of pregnant women with IDA were 8.92 ± 1.46 g/dL and 12.63 ± 1.36 µg/L, respectively. The mean cord blood hemoglobin and serum ferritin levels were 15.24 ± 1.04 g/dL and 27.79 ± 4.37 µg/L, respectively. Only 15.18% neonates born from mother with IDA were anemic (Hb &lt;14 g/dL).

Effect of warming different intravenous fluids on maternal and neonatal outcomes during cesarean section - comparison of crystalloids and colloids

Background/Aim: Fluid warming is a useful method to prevent maternal and neonatal hypothermia. Because colloids stay in the intravascular space longer than crystalloids, their protective effect against hypothermia may be more emphasized. The aim of this study is to compare the effects of using warmed colloids or crystalloids on maternal and neonatal core temperatures, neonatal blood gas values, and maternal thermal comfort scores. Methods: After ethical approval, 220 ASA I or II pregnant women, scheduled for cesarean section with spinal anesthesia were enrolled in the study. Patients were assigned to receive hydroxyethyl starch (group HES) or Ringer’s lactate solution (group RL) throughout the intraoperative study period. Once the patient entered the room, fluids were actively warmed with fluid warmer to 41 °C in both groups. Measurement of maternal core temperature (MT) and thermal comfort score (TCS) were performed before starting intravenous fluid administration (control), at the time of delivery of the neonate (delivery), 15th, 30th minutes, and at the end of the surgery. Tympanic temperature of the neonates was measured 1 minute after delivery. Blood gas samples from the umbilical artery of the neonates at the 1st minute and Apgar scores at 1st and 5th minutes after delivery were evaluated. Results: Maternal tympanic temperatures, maternal thermal comfort scores were higher at all measurement values other than control measurement in group HES. Neonatal tympanic temperatures (P=0.051), neonatal umbilical artery cord pH (P

Relationship between umbilical cord practices and neonatal cord outcomes among neonates attending Nakuru Level Five County and Referral Hospital, Nakuru, Kenya

Background: Globally neonatal mortality rate is 22 per 1000 live births, 98% of these deaths are from developing countries and umbilical infections contribute significantly to the etiology of these deaths. Methods: A hospital based unmatched case control study design with study population comprising of neonates 14 days and older. Cases were neonates with umbilical granuloma, delayed cord separation and omphalitis. Controls were neonates without the aforementioned conditions. Collection of data was done using interviewer administered questionnaire and analyses done using SPSS Version 26. Probability outcomes relative to the predictors were predicted using logistic regression. Results: Prevalence of adverse cord outcomes was 56.60%. 14.72% umbilical granuloma, 17.07% delayed cord separation and 29.52% omphalitis. Use of chlorhexidine digluconate for cord care was statistically significant in protecting an adverse cord outcome (OR=0.070, p=0.000) while use of other substances for cord care was statistically significant in predisposing the newborn to an adverse cord outcome (OR=11.813, p=0.000). Multiparity (OR=0.666, p=0.353) and sponge bathing the baby (OR=0.618, p=0.264) were protective while primiparity (OR=1.666, p= 0.241) and bathing the baby by immersion (OR=1.573, p=0.297) were risk factors. Conclusions: Prevalence of adverse cord outcomes (umbilical granuloma, delayed cord separation, omphalitis) was high compared to the global prevalence thus a major public health concern. Adoption of WHO evidenced based chlorhexidine digluconate policy should be encouraged as a universally accepted method of cord care. The practice of home based post natal visits by community health workers should be strengthened as a way to link facility recommendations to the home environment

Effect of Maternal Catalase Supplementation on Reproductive Performance, Antioxidant Activity and Mineral Transport in Sows and Piglets.

This experiment was conducted to investigate the effects of maternal catalase (CAT) supplementation on reproductive performance, antioxidant enzyme activities, mineral transport, and mRNA expression of related genes in sows and offspring. A total of 40 pregnant sows at 95 days of gestation with similar parity (3-5 parities) and back-fat thickness were assigned randomly and equally into the control (CON) group (fed a basal diet) and CAT group (fed a basal diet supplemented with 660 mg/kg CAT; CAT activity, 280 U/g). The reproductive performance was recorded, and the placenta and blood samples of sows and neonatal piglets, as well as the jejunum and ileum samples from neonatal boars (eight replicates per group), were collected. Results showed that dietary supplementation with CAT significantly decreased the intrauterine growth restriction (IUGR) rate and increased the activity of serum CAT in neonatal piglets and umbilical cords (p &lt; 0.05). In addition, CAT supplementation tended to improve total antioxidant capacity (T-AOC) levels in the maternal serum (p = 0.089) and umbilical cords of piglets (p = 0.051). The serum calcium (Ca), manganese (Mn), and zinc (Zn) of farrowing sows and Mn concentration in the umbilical cord, and serum Ca, magnesium (Mg), copper (Cu), and Mn of neonatal piglets were significantly increased (p &lt; 0.05) in the CAT group. CAT supplementation downregulated mRNA expression of TRPV6 and CTR1 (p &lt; 0.05), Cu/Zn SOD (p = 0.086) in the placenta and tended to increase the mRNA expression of the glutathione peroxidase 1 (GPX1) (p = 0.084), glutathione peroxidase 4 (GPX4) (p = 0.063), and CAT (p = 0.052) genes in the ileum of piglets. These results showed that the maternal CAT supplementation improved fetal growth by decreasing the IUGR rate, and modulated antioxidant activity, as well as mineral elements in the pregnant sows and their piglets.

Disentangling the effects of restriction and exchange with diffusion exchange spectroscopy.

Diffusion exchange spectroscopy (DEXSY) is a multidimensional NMR technique that can reveal how water molecules exchange between compartments within heterogeneous media, such as biological tissue. Data from DEXSY experiments is typically processed using numerical inverse Laplace transforms (ILTs) to produce a diffusion-diffusion spectrum. A tacit assumption of this ILT approach is that the signal behavior is Gaussian - i.e., the spin echo intensity decays exponentially with the degree of diffusion weighting. The assumptions that underlie Gaussian signal behavior may be violated, however, depending on the gradient strength applied and the sample under study. We argue that non-Gaussian signal behavior due to restrictions is to be expected in the study of biological tissue using diffusion NMR. Further, we argue that this signal behavior can produce confounding features in the diffusion-diffusion spectra obtained from numerical ILTs of DEXSY data - entangling the effects of restriction and exchange. Specifically, restricted signal behavior can result in broadening of peaks and in the appearance of illusory exchanging compartments with distributed diffusivities, which pearl into multiple peaks if not highly regularized. We demonstrate these effects on simulated data. That said, we suggest the use of features in the signal acquisition domain that can be used to rapidly probe exchange without employing an ILT. We also propose a means to characterize the non-Gaussian signal behavior due to restrictions within a sample using DEXSY measurements with a near zero mixing time or storage interval. We propose a combined acquisition scheme to independently characterize restriction and exchange with various DEXSY measurements, which we term Restriction and Exchange from Equally-weighted Double and Single Diffusion Encodings (REEDS-DE). We test this method on ex vivo neonatal mouse spinal cord - a sample consisting primarily of gray matter - using a low-field, static gradient NMR system. In sum, we highlight critical shortcomings of prevailing DEXSY analysis methods that conflate the effects of restriction and exchange, and suggest a viable experimental approach to disentangle them.

Toll-like receptors 2 and 4 differentially regulate the self-renewal and differentiation of spinal cord neural precursor cells

BackgroundToll-like receptors (TLRs) represent critical effectors in the host defense response against various pathogens; however, their known function during development has also highlighted a potential role in cell fate determination and neural differentiation. While glial cells and neural precursor cells (NPCs) of the spinal cord express both TLR2 and TLR4, their influence on self-renewal and cell differentiation remains incompletely described.MethodsTLR2, TLR4 knock-out and the wild type mice were employed for spinal cord tissue analysis and NPCs isolation at early post-natal stage. Sox2, FoxJ1 and Ki67 expression among others served to identify the undifferentiated and proliferative NPCs; GFAP, Olig2 and β-III-tubulin markers served to identify astrocytes, oligodendrocytes and neurons respectively after NPC spontaneous differentiation. Multiple comparisons were analyzed using one-way ANOVA, with appropriate corrections such as Tukey's post hoc tests used for comparisons.ResultsWe discovered that the deletion of TLR2 or TLR4 significantly reduced the number of Sox2-expressing NPCs in the neonatal mouse spinal cord. While TLR2-knockout NPCs displayed enhanced self-renewal, increased proliferation and apoptosis, and delayed neural differentiation, the absence of TLR4 promoted the neural differentiation of NPCs without affecting proliferation, producing long projecting neurons. TLR4 knock-out NPCs showed significantly higher expression of Neurogenin1, that would be involved in the activation of this neurogenic program by a ligand and microenvironment-independent mechanism. Interestingly, the absence of both TLR2 and TLR4, which induces also a significant reduction in the expression of TLR1, in NPCs impeded oligodendrocyte precursor cell maturation to a similar degree.ConclusionsOur data suggest that Toll-like receptors are needed to maintain Sox2 positive neural progenitors in the spinal cord, however possess distinct regulatory roles in mouse neonatal spinal cord NPCs—while TLR2 and TLR4 play a similar role in oligodendrocytic differentiation, they differentially influence neural differentiation.

RHD exon 5, 7 and 10 targeted non-invasive prenatal screening of fetal Rhesus-D (RhD) in selected RhD negative pregnant women in Ethiopia.

BackgroundA majority of non-invasive prenatal screening studies determining fetal RhD status have been tested on Caucasian and Asian populations, but limited or no studies have been conducted on the Ethiopian population. In the current study, we carried non-invasive prenatal screening of fetal RHD genotype in selected RhD negative Ethiopian pregnant women.MethodsCell-free DNA was extracted from the plasma samples of 117 RhD pregnant women between 9 and 38 weeks of gestation. Fetal RHD genotypes were detected by targeting exons 5, 7 and 10 of the RHD gene by using real-time PCR assay. RHD genotypic results were confirmed by neonatal cord blood serology.ResultsFetal RHD genotyping was conclusive in all 117 subjects. RHD genotype was correctly predicted in 115 of 117 cases, thus the test yielded 98.3% accuracy (95%CI: 97.3–99.1%). Among 115 cases, 105 were genotyped as RHD positive and 12 were genotyped as RHD negative. The sensitivity and specificity of the test were 99.1% (95% CI: 94.8–99.9%) and 91.7% (95%CI: 61.5–99.7%) respectively. The negative and positive predictive values were 99.9% (95%CI: 99.2–99.9%) and 54.0% (95% CI: 15.2–88.4%) respectively. SRY genotyping results were in complete concordance with fetal sex.ConclusionMulti exon targeted non-invasive prenatal screening test for fetal RhD determination exhibited high accuracy and sensitivity. A confirmatory study with a bigger size of study subjects is warranted before enabling clinical implementation.

Fetal sex and maternal fasting glucose affect neonatal cord blood-derived endothelial progenitor cells

BackgroundMaternal cardiovascular risk factors (CVRF) in pregnancy, i.e., obesity and hyperglycemia, transmit to the fetus and affect placental and fetal endothelial function. Moreover, a sex dimorphism in endothelial function and susceptibility towards CVRF exists already in utero. Endothelial colony-forming cells (ECFC) are circulating endothelial progenitors highly present in neonatal cord blood and sensitive to CVRF. This study investigated whether fetal sex or subtle maternal metabolic changes within healthy range alter fetal ECFC outgrowth.MethodsOutgrowth of ECFC from cord blood of male (n = 31) and female (n = 26) neonates was analyzed after healthy pregnancies and related to fetal sex and maternal metabolic parameters.ResultsMale ECFC grew out earlier (−20.57% days; p = 0.031) than female. Although all women were non-diabetic, higher levels of fasting plasma glucose (FPG) at midpregnancy increased the time required for colony outgrowth (OR: 1.019; p = 0.030), which, after stratifying for fetal sex, was significant only in the males. Gestational weight gain and BMI did not affect outgrowth. Colony number was unchanged by all parameters.ConclusionsFetal sex and maternal FPG within normal range alter ECFC function in utero. A role of ECFC in postnatal angiogenesis and vasculogenesis has been suggested, which may be affected by altered outgrowth dynamics.ImpactThis study is the first to report that a sexual dimorphism exists in ECFC function, as cells of female progeny require a longer period of time until colony outgrowth than ECFC of male progeny.Our data show that ECFC function is highly sensitive and affected by maternal glucose levels even in a normal, non-diabetic range.Our data raise the question of whether maternal plasma glucose in pregnancy should be considered to play a critical role even in the non-diabetic setting.

Comparison of phenylephrine and norepinephrine for treatment of spinal hypotension during elective cesarean delivery- A randomised, double-blind study.

Hypotension following subarachnoid block for cesarean delivery (CD) is common. We compared the effect of bolus administration of norepinephrine and phenylephrine on umbilical artery pH (primary objective) and their efficacy for the treatment of maternal hypotension (secondary objective) in term parturients undergoing elective CD under spinal anesthesia. In a randomized, double-blinded study, parturients received 1 mL boluses of either phenylephrine 100 μg/mL (group phenylephrine; n = 45) or norepinephrine 7.5 μg/mL (group norepinephrine; n = 45) whenever maternal systolic blood pressure decreased to ≤80% of baseline. Maternal hemodynamic changes, vasopressor, and atropine requirement and neonatal outcome (umbilical cord blood gas analysis, Apgar scores, neonatal neurobehavioral response) were assessed. The Apgar scores and umbilical cord blood gas analysis were comparable between groups. The neurobehavioral scale score was significantly higher in group NE compared with that in group PE at 24 h and 48 h; P = 0.007 and 0.002, respectively. The number of vasopressor doses and time to the first vasopressor requirement for maintaining systolic pressure >80% of baseline was comparable in both groups. Incidence of bradycardia (P = 0.009), reactive hypertension (P = 0.003), and dose requirement of atropine (P = 0.005) was higher in group PE compared with group NE. In term normotensive parturients who received bolus norepinephrine 7.5 μg or phenylephrine 100 μg for the treatment of post-spinal hypotension during CD, neonatal umbilical cord blood gas analysis and Apgar scores were comparable. Norepinephrine use was associated with a lower incidence of maternal bradycardia and reactive hypertension compared with phenylephrine.

Maternal caffeine intake and DNA methylation in newborn cord blood

Epigenetic mechanisms may underlie associations between maternal caffeine consumption and adverse childhood metabolic outcomes. However, limited studies have examined neonate DNA methylation (DNAm) patterns in the context of preconception or prenatal exposure to caffeine metabolites. We examined preconception and pregnancy caffeine exposure with DNAm alterations in neonate cord blood (n=378). In a secondary analysis of the Effects of Aspirin in Gestation and Reproduction Trial (EAGeR), we measured maternal caffeine, paraxanthine, and theobromine concentrations from stored serum collected preconception (on average 2 months before pregnancy) and at 8 weeks of gestation. In parallel, self-reported caffeinated beverage intake was captured via administration of questionnaires and daily diaries. We profiled DNAm from the cord blood buffy coat of singletons using the MethylationEPIC BeadChip. We assessed associations of maternal caffeine exposure and methylation β values using multivariable robust linear regression. A false discovery rate (FDR) correction was applied using the Benjamini-Hochberg method. In preconception, the majority of women reported consuming 1 or fewer servings/day of caffeine on average, and caffeine and paraxanthine metabolite levels were 88 and 36µmol/L, respectively. Preconception serum caffeine metabolites were not associated with individual cytosine-guanine (CpG) sites (FDR>5%), though pregnancy theobromine was associated with DNAm at cg09460369 near RAB2A (β = 0.028; SE=0.005; FDR P=0.012). Preconception self-reported caffeinated beverage intake compared to no intake was associated with DNAm at cg09002832 near GLIS3 (β = -0.013; SE=0.002; FDR P=0.036). No associations with self-reported intake during pregnancy were found. Few effects of maternal caffeine exposure on neonate methylation differences in leukocytes were identified in this population with relatively low caffeine consumption.

DDAH1 SNP rs480414 that protects against the development of pulmonary hypertension in bronchopulmonary dysplasia results in lower nitric oxide production in neonatal cord blood-derived lymphoblastoid cell lines.

Bronchopulmonary dysplasia (BPD) is chronic lung disease of prematurity and pulmonary hypertension (PH) is a major contributor to morbidity and mortality in BPD patients. Nitric oxide (NO) is a vasodilator and apoptotic mediator made by NO synthase (NOS). NOS is inhibited by asymmetric dimethylarginine (ADMA), and dimethylarginine dimethylaminohydrolase (DDAH) hydrolyzes ADMA. Previously, in a BPD patient cohort, we identified single nucleotide polymorphism (SNP) DDAH1 rs480414 (G > A) that was protective against developing PH. This study aims to determine functional consequences of the DDAH1 SNP in lymphoblastoid cell lines (LCLs) derived from neonatal cord blood. We tested the hypothesis that DDAH1 SNP (AA) results in DDAH1 gain of function, leading to greater NO-mediated apoptosis compared to DDAH1 wild-type (GG) in LCLs. LCLs were analyzed by Western blot (DDAH1, cleaved and total caspase-3 and -8, and β-actin), and RT-PCR (DDAH1, iNOS). Cell media assayed for nitrites with chemiluminescence NO analyzer, and conversion of ADMA to L-citrulline was measured by spectrophotometry. LCLs with DDAH1 SNP had similar levels of DDAH1 protein and mRNA expression, as well as DDAH activity, compared to DDAH1 WT LCLs. There were also no changes in cleaved caspase-3 and -8 protein levels. LCLs with DDAH1 SNP had similar iNOS mRNA expression. Nitrite levels in media were lower for DDAH1 SNP LCLs compared to DDAH1 WT LCLs (p < 0.05). Contrary to our hypothesis, we found that NO production was lower in DDAH1 SNP LCLs, indicative of a loss of function phenotype.

Lipoxins in the Nervous System: Brighter Prospects for Neuroprotection.

Lipoxins (LXs) are generated from arachidonic acid and are involved in the resolution of inflammation and confer protection in a variety of pathological processes. In the nervous system, LXs exert an array of protective effects against neurological diseases, including ischemic or hemorrhagic stroke, neonatal hypoxia-ischemia encephalopathy, brain and spinal cord injury, Alzheimer’s disease, multiple sclerosis, and neuropathic pain. Lipoxin administration is a potential therapeutic strategy in neurological diseases due to its notable efficiency and unique superiority regarding safety. Here, we provide an overview of LXs in terms of their synthesis, signaling pathways and neuroprotective evidence. Overall, we believe that, along with advances in lipoxin-related drug design, LXs will bring brighter prospects for neuroprotection.

Mapping the Cord Blood Transcriptome of Pregnancies Affected by Early Maternal Anemia to Identify Signatures of Fetal Programming.

ContextAnemia during early pregnancy (EP) is common in developing countries and is associated with adverse health consequences for both mothers and children. Offspring of women with EP anemia often have low birth weight, which increases risk for cardiometabolic diseases, including type 2 diabetes (T2D), later in life.ObjectiveWe aimed to elucidate mechanisms underlying developmental programming of adult cardiometabolic disease, including epigenetic and transcriptional alterations potentially detectable in umbilical cord blood (UCB) at time of birth.MethodsWe leveraged global transcriptome- and accompanying epigenome-wide changes in 48 UCB from newborns of EP anemic Tanzanian mothers and 50 controls to identify differentially expressed genes (DEGs) in UCB exposed to maternal EP anemia. DEGs were assessed for association with neonatal anthropometry and cord insulin levels. These genes were further studied in expression data from human fetal pancreas and adult islets to understand their role in beta-cell development and/or function.ResultsThe expression of 137 genes was altered in UCB of newborns exposed to maternal EP anemia. These putative signatures of fetal programming, which included the birth weight locus LCORL, were potentially mediated by epigenetic changes in 27 genes and associated with neonatal anthropometry. Among the DEGs were P2RX7, PIK3C2B, and NUMBL, which potentially influence beta-cell development. Insulin levels were lower in EP anemia–exposed UCB, supporting the notion of developmental programming of pancreatic beta-cell dysfunction and subsequently increased risk of T2D in offspring of mothers with EP anemia.ConclusionsOur data provide proof-of-concept on distinct transcriptional and epigenetic changes detectable in UCB from newborns exposed to maternal EP anemia.

Human menstrual blood-derived stem cells combined with a new 3D bioprinted composite scaffold for spinal cord injury treatment

Spinal cord injury (SCI) regeneration and repair is a worldwide medical issue, and stem cell therapy is an emerging treatment strategy. Previously, our research group found that transplanting menstrual blood-derived mesenchymal stem cells (MenSCs) can promote axon regeneration in rats SCI. However, the SCI pathological microenvironment inhibits stem cell survival after transplantation. The acellular spinal cord is the most similar to the spinal cord in terms of three-dimensional structure and natural biological activity, and it can efficiently regulate axon growth and promote synapse formation. Based on recent progress in the understanding of scar-free spinal cord repair in neonatal mice, it was postulated that in the acellular spinal cord microenvironment of neonatal rats, it would be more active ingredients to promote axon regeneration and inhibit glial scars, than in the adult rat. The hypothesis was formulated that a new 3D bioprinted scaffold with neonatal acellular spinal cord/GelMA hydrogel/MenSCs would efficiently repair SCI. The neonatal acellular spinal cord can provide a dual bionic microenvironment for axon regeneration, optimizing bioactive composition and microstructure, and the GelMA hydrogel guarantees the scaffold's mechanical properties. Then, using 3D bioprinting technology, the pore size of the scaffold can be precisely controlled, and the spinal cord conduction tract path may be stimulated. Moreover, a tissue-engineered spinal cord can be constructed by planting MenSCs in the pores. Overall, our hypothesis suggests that 3D bioprinted neonatal acellular spinal cord/GelMA hydrogel composite scaffold coupled with MenSCs might have more beneficial therapeutic effects on spinal cord injury rehabilitation.