Placental Research Articles

The Fetal Effect of Maternal Caffeine Consumption During Pregnancy—A Review

Caffeine is commonly used to excess by the general public, and most pregnant women drink caffeine on a daily basis, which can become a habit. Maternal caffeine intake during pregnancy is associated with severe gestational outcomes. Due to its lipophilic nature, caffeine can cross the blood–brain barrier, placental barrier, and even amniotic fluid. It can be found in substantive amounts in breast milk and semen. There has been a reported drop in neonatal anthropometric measurements with increased caffeine consumption in some cohort studies. This narrative review using literature titles and abstracts from the electronic databases of PubMed, Embase, and Scopus investigates the data linking maternal caffeine use to unfavorable pregnancy outcomes. It also evaluates the validity of the recommendations made by health professionals on caffeine consumption by mothers from the available literature. The results of our comprehensive literature search of case–control studies, cohort studies, randomized control trials, and meta-analyses, imply that caffeine use during pregnancy is linked to miscarriage, stillbirth, low birth weight, and babies that are small for gestational age. It was also found that there may be effects on the neurodevelopment of the child and links to obesity and acute leukemia. These effects can even be seen at doses well below the daily advised limit of 200 mg. The genetic variations in caffeine metabolism and epigenetic changes may play a role in the differential response to caffeine doses. It is crucial that women obtain solid, evidence-based guidance regarding the possible risks associated with caffeine.

A comprehensive review of the roles of T-cell immunity in preeclampsia

Preeclampsia (PE) is an obstetrical disorder that occurs after the 20th week of gestation. It is recognized as one of the “Great Obstetrical Syndromes” and principally contributes to maternal morbidity and mortality. PE has been associated with a range of immune disorders, including a preponderance of T helper (Th) 1 over Th2 cells and imbalanced levels of Th17 and T regulatory cells (Tregs). During pregnancy, T cells safeguard the placenta against immune rejection and aid embryo implantation while involved in pregnancy complications, such as PE. Promoting alloantigen-specific Treg cells is a potential preventive and therapeutic strategy for PE. However, ensuring the safety of mothers and infants is of the utmost importance since the risk-benefit ratio of reproductive and obstetric conditions differs significantly from that of immune diseases that pose a life-threatening risk. In this review, we systematically summarize the roles of T-cell immunity in the peripheral blood, reproductive tissues, and at the maternal-fetal interface of PE patients. Furthermore, the recent therapeutic approaches centered on targeting T cell immunity in PE are critically appraised.

Carbonic anhydrase IV deficiency causes intrauterine embryonic loss in mice.

Members of the carbonic anhydrase gene family, responsible for the reversible hydration of carbon dioxide, participate in several important biological processes including processes involved in fertilization. CAIV has been shown to play a role in sperm cell capacitation and regulation of sperm motility and is present in mature murine placentae. The present study specifically analyses the distribution of CAIV in female reproductive organs and during early placenta development. Immunostaining for CAIV was performed on female reproductive organs (ovary, fallopian tube, uterus, vagina) of non-pregnant mice and on implantation sites of early pregnancy between 4.5 and 9.5 days post coitum (dpc). Sex typing of embryos was performed by PCR using three separated gene combinations for X- and Y-chromosomes, respectively. Additionally, reproductive outcome of CAIV-deficient mice was determined. CAIV is largely absent in the female reproductive organs of non-pregnant mice. Immunostaining for CAIV was present in the blastocyst and in consecutive stages of the developing embryo. In the endometrial epithelium distant from the implantation chamber, CAIV is induced from 8.5 dpc onwards. Moreover, the yolk sac epithelium, the trophoblast giant cells and the labyrinthine compartment of the developing hemochorial placenta shows a strong immunostaining for CAIV. In heterozygous mating, the number of CAIV knockout pups is significantly reduced than was to be expected according to the mendelian rules, while homozygous mating of CAIV knockout mice results in a significant reduction of litter size which is mainly due to a reduced number of female mice born. Since at 9.5 dpc the number of female embryos is rather higher than that of males, the observed reduction of female offspring appears to be due to a defect in placentation after 9.5 dpc. Thus, CAIV seems to be involved in the signaling network of embryo development, implantation, and placentation.

Upregulation of Autophagy During the Differentiation of Primary Human Term Cytotrophoblast Cells into Syncytial Cells: Ultrastructural Analysis

The villous trophoblast cells are of fundamental importance because they fulfill a variety of functions that are vital for the growth of the fetus and the maintenance of pregnancy. A simple in vitro villous trophoblast cell model that grows on standard tissue culture plates has been utilized for various functional studies on villous trophoblast cells. Despite the potential value of incorporating electron microscopy analysis in reports on functional analysis of primary human trophoblast cells, electron microscopy analysis is exclusively ancillary to functional analysis in previous publications. In the context of autophagy research of villous trophoblast cells using primary trophoblast cells, a detailed ultrastructural analysis of autophagy flux using electron microscopy is imperative; however, it has not been conducted to date. In this study, we isolated term villous trophoblast cells (i.e., cytotrophoblast cells, CTB cells) using the most up-to-date isolation method for isolating pure CTB cells from human term placenta and investigated the ultrastructural dynamic process of autophagy of cultured CTB cells by means of transmission electron microscopy. The initial 6 h culture resulted in CTB cell aggregation; however, the majority of CTB cells did not differentiate into syncytial cells. In contrast, after 72 h, CTB cells exhibited a promotion of differentiation into syncytial cells. The electron microscopy analysis revealed the upregulation of autophagy and visualized unique autophagic profiles during differentiation into syncytial cells, which exhibited perinuclear accumulation of extremely large autophagosomes/autolysosomes. This study provides novel insights into the reproductive biology of primary trophoblast cells, thereby demonstrating the substantial value of primary trophoblast cells as research resources.

Placental Adaptation to Hypoxia: The Case of High-Altitude Pregnancies

Even in the highest inhabited regions of the world, well above 2500 m altitude, women become pregnant and give birth to healthy children. The underlying adaptation to hypobaric hypoxia provides interesting insights into the physio(patho)logy of the human placenta. Although increasing altitude is regularly associated with fetal growth restriction (FGR), oxygen deficiency does not appear to be a direct cause. Rather, placental oxygen consumption is reduced to maintain the oxygen supply to the fetus. This comes at the expense of placental synthesis and transport functions, resulting in inappropriate nutrient supply. The hypoxia-inducible factor (HIF-1α), which modulates the mitochondrial electron transport chain to protect placental tissue from reactive oxygen species, plays a key role here. Reduced oxygen consumption also reflects decreased placental vascularization and perfusion, which is accompanied by an increased risk of maternal pre-eclampsia at high altitude. In native highlanders, the latter seems to be attenuated, partly due to a lower release of HIF-1α. In addition, metabolic peculiarities have been described in indigenous people that enhance glucose availability and thus reduce the extent of FGR. This review attempts to revisit the (albeit incomplete) knowledge in this area to draw the clinical reader’s attention to the crucial role of the placenta in defending the fetus against hypoxia.

Ferritin as a biomarker of multiple organ pathology: anti-inflammatory and antioxidant peptides of standardized human placenta hydrolysate

Ferritin as a biomarker of multiple organ pathology: anti-inflammatory and antioxidant peptides of standardized human placenta hydrolysate

A novel virulent core genome multilocus sequence type CT 11424 of Listeria monocytogenes isolate causing stillbirth in Bangladesh

BackgroundListeria monocytogenes is a foodborne pathogen that can lead to severe pregnancy outcomes. This study reports the clinical and genomic characteristics of a Listeria-mediated stillbirth identified in January 2022 through the Child Health and Mortality Prevention Surveillance (CHAMPS) project in Bangladesh. The Lm-BD-CHAMPS-01 isolate was recovered from the blood and cerebrospinal fluid (CSF) of a male stillborn. Maternal history, clinical, and demographic data were collected by the CHAMPS surveillance platform. An expert panel evaluated all reports to determine the role of L. monocytogenes infection in the causal chain of stillbirth. Genomic characterization included multilocus sequence typing (MLST), core genome MLST (cgMLST), serotyping, and the presence or absence of virulence genes. Genetic divergence and phylogenetic analyses were conducted to determine the relationship with other reported isolates globally.ResultsThe isolate Lm-BD-CHAMPS-01 was identified as a novel cgMLST CT11424. It belonged to ST 308, Serotype 4b, Clonal Complex 1, and Phylogenetic Lineage 1. Key L. monocytogenes virulence genes facilitating the crossing of the placental barrier, including full-length inlA, LIPI-1, and LIPI-3, were detected. The isolate was closely related to clinical L. monocytogenes isolates, as determined by GrapeTree based on cgMLST. SNP-based phylogenetic analysis found Lm-BD-CHAMPS-01 to be the most distant from other CC1 isolates in the database. Possible sources of infection included the consumption of contaminated raw vegetables or exposure to pigeons.ConclusionsThis is the first genome sequence of clinical L. monocytogenes from Bangladesh, which also caused stillbirth. Rural healthcare professionals should be aware of L. monocytogenes infection risks during pregnancy. Pregnant women should be counseled on the dangers of exposure to animals or birds and consumption of potentially contaminated raw food to prevent adverse pregnancy outcomes due to L. monocytogenes infection.

Preventive vs. therapeutic effects of Shoutai Wan: Maintaining an acidic microenvironment at the maternal-fetal interface to promote angiogenesis and minimize pregnancy loss in RSA mice.

The classic TCM prescription, Shoutai Wan (STW), is extensively used in clinical settings to manage threatened miscarriage and Recurrent spontaneous abortion (RSA). The complexity of pregnancy physiology, coupled with diverse etiologies, and the specificity of energy metabolism for normal embryo attachment and development, pose challenges to clinical diagnosis and treatment. The specific molecular mechanisms of how STW regulates these biological processes and contributes to the treatment of RSA remain to be elucidated. This study aims to investigate the causes of early pregnancy loss in RSA mice and explore how STW mitigates this loss. An RSA mouse model will be established and treated with STW and Dydrogesterone (DYD). Embryo loss will be quantified on the 14th day of pregnancy, and embryos will be collected on the 6th and 10th days to observe the embryonic condition and assess pathological changes. The study will analyze aerobic glycolysis and angiogenesis at the maternal-fetal interface (MFI). Additionally, STW on a knockdown LDHA mouse model and Human Endometrial Microvascular Endothelial Cells (HEMECs) in vitro will also be examined to verify the mechanism. Compared with the control group, the RSA group exhibited significant embryo loss, and reduced levels of aerobic glycolysis at the MFI, the precarious acidic microenvironment (AME), and the PI3K/AKT/mTOR signaling axis downregulated, leading to impaired angiogenesis, which ameliorated following STW treatment. STW treatment enhanced key aerobic glycolysis enzymes-HK2, PKM2, LDHA-and lactate levels, thereby maintaining the AME and upregulating the PI3K/AKT/mTOR axis. This, in turn, promoted the expression of angiogenesis-related factors (VEGFA and VEGFR2) at the MFI, thereby improving angiogenesis, and the same was seen in sh-LDHA mice. In vitro studies confirmed that STW could counteract the glycolysis decline caused by increased oxygen levels, a recovery that was impaired after LDHA knockdown or PI3K inhibition. In RSA mice, disturbances in aerobic glycolysis at the MFI prevent the maintenance of a stable AME, thus impairing angiogenesis and leading to embryo loss, and STW effectively improve early pregnancy outcomes, and laying the foundation for uterine spiral artery remodeling.

Aneurysm clipping on WEB device: A feasibility study using a human ex-vivo aneurysm model.

The Woven EndoBridge (WEB) device is a breakthrough in brain aneurysm treatment, widely used for intracranial wide neck and complex aneurysms, especially at bifurcations. Although effective, aneurysm recurrence can occur post-treatment and the impact of clipping angle on the WEB mesh remains unclear. In this study, using a human placenta-based ex-vivo aneurysm model, we simulated the application of aneurysm clipping on WEB-treated aneurysms to elucidate the technical aspects and nuances, particularly the influence of the angle of clip application on WEB collapse. Human placentas were used to create aneurysms by suture-closing of the placenta vessels. The WEB devices were positioned in the aneurysms under fluoroscopic guidance, and different clip positions/angles on the WEB device were tested. Imaging was used to monitor the effects of clipping on the WEB device and aneurysm. All clipping positions achieved complete aneurysm occlusion, collapsing the WEB mesh with no flow observed into the aneurysm. Despite the WEB device's displacement within the sac, no significant morphological changes to the aneurysm were identified, and the presence of a neck remnant did not affect the adequacy of blade apposition. Microsurgical clipping reliably collapses the WEB device, regardless of its placement or orientation. Our model suggests that clipping can effectively address in-vivo recurrences and persistent aneurysm filling of aneurysms treated with the WEB device. The model does not assess the influence of WEB healing within the aneurysm. Operators should be aware of possible WEB shape changes when clipping its mesh.

Extracellular vesicles in ZIKV infection: Carriers and facilitators of viral pathogenesis?

Zika virus (ZIKV) is a flavivirus of significant epidemiological importance, utilizing various transmission strategies and infecting "immune privileged tissues" during both the pre- and postnatal periods. One such transmission method may involve extracellular vesicles (EVs). EVs can travel long distances without degrading, carrying complex messages that trigger different responses in recipient cells. They can easily cross specialized tissue barriers, such as the placental barrier and the blood-brain barrier, which protects the central nervous system. It is known that some viruses can hijack and exploit the EVs biogenesis machinery to package regulatory elements, viral segments, and even complete viral genomes. This allows them to evade the immune system, amplify their tropism, and enhance their spread. ZIKV likely uses EVs produced by infected cells to insert its genomic RNA or parts of it. This mechanism can ensure viral entry and infection of the nervous tissue, partly explaining its broad viral tropism and silent persistence in various tissues and organs for months. This narrative review summarizes the main features of ZIKV and EVs, highlighting the most recent evidence on the involvement and effects of EVs during ZIKV infection. It also discusses the possibility of EVs acting as carriers of ZIKV through the nervous tissue.

Deletion of BMP4 impairs trophoblast function and decidual macrophage polarization via autophagy leading to recurrent spontaneous abortion.

Bone morphogenetic protein 4 (BMP4) is widely involved in the regulation of cell proliferation and differentiation, but its role in Recurrent Spontaneous Abortion (RSA) remains unclear. RSA is a disease that affects roughly 1-2% of partner pairs, but its pathogenesis is still unclear. In recent years, many studies have focused on the role of decidual macrophages in RSA. In this study, we found decreased expression levels of BMP4 in villous tissues of RSA patients and found that low expression of RUNX2 leads to down-regulation of BMP4, which impairs trophoblast function. More importantly, we found in both co-culture system and human recombinant BMP4 protein models that BMP4 overexpression polarizes THP-1-derived macrophages toward M2, and down-regulation of BMP4 leads to macrophage polarization toward M1. Mechanically, we found that BMP4 promotes macrophage polarization via regulating autophagy level. The recovery experiment was further confirmed that 3-MA (autophagy inhibitor) inhibit THP-1-derived macrophage polarization toward M2 induced by BMP4 overexpression and exogenous addition of rBMP4, and rapamycin (autophagy agonists) inhibit macrophages polarization toward M1 from down-regulation of BMP4. Our study further reveals the mechanism of maternal-fetal interface cell interactions in RSA, which can help in the diagnosis and treatment of RSA.

Compromised PPARγ Mediated Impaired Placental Glucose Transport via the PI3K/AKT Signaling Pathway is Associated with FGR.

Fetal growth restriction (FGR) is a condition in which a fetus cannot grow to its full potential during pregnancy. It is a leading cause of perinatal mortality and morbidity. However, the underlying etiology remains elusive. Here, we report that peroxisome proliferator-activated receptor γ (PPARγ) is inactivated in the trophoblasts of human placenta of FGR-complicated pregnancies. In the FGR placentas, p-PI3KTyr458 and p-AKTSer473 levels were also lowered. Additionally, there was a reduction in GLUT3 and GLUT4 levels in the cell membrane. Consistently, FGR patients showed decreased glucose concentrations in both the placenta and umbilical cord blood compared to that in normal pregnancy. In mouse models, deletion of PPARγ in trophoblasts and RUPP surgery successfully induced FGR and replicated these changes. Modulating PPARγ activity using rosiglitazone or GW9662 in BeWo cells resulted in the activation or inhibition of the PI3K/AKT signaling pathway, as well as the promotion or reduction of membrane translocation of GLUT3 and GLUT4, ultimately affecting glucose uptake in trophoblast cells. MK-2206 blocked these regulatory effects of rosiglitazone in BeWo cells. Furthermore, the administration of rosiglitazone encapsulated in placenta-targeting nanoparticles improved the growth and development of fetal mice in the RUPP group. In summary, PPARγ in trophoblast cells orchestrates the translocation of GLUT3 and GLUT4 to the cellular membrane via the PI3K/AKT signaling pathway, thereby regulating cellular glucose uptake and transport. Dysfunctions in this mechanism are strongly associated with FGR. Therefore, targeted activation of PPARγ in the placenta may be a potentially efficacious intrauterine intervention for FGR.

Role of calcium overload-mediated disruption of mitochondrial dynamics in offspring neurotoxicity due to methylmercury exposure during pregnancy and lactation.

Methylmercury (MeHg) is a potent neurotoxicant with neurodevelopmental toxicity that is widely ingested into the body through drinking water and food. MeHg crosses the placental barrier and accumulates in the brain of the fetus, affecting the growth and development of the central nervous system. Although it has been demonstrated that MeHg induces neuronal calcium overload in the rat cerebral cortex, the role of calcium overload in MeHg-induced neurodevelopmental toxicity remains unclear. Here, we used ICR-pregnant mice and their resulting offspring and administered the BAPTA-AM calcium antagonist to investigate the molecular mechanisms by which MeHg exposure during gestation and lactation affects neurodevelopment. We found that exposure to MeHg during gestation and lactation resulted in developmental arrest and neurobehavioral dysfunction in the offspring, with calcium overload, disturbed mitochondrial dynamics, and apoptosis. However, the calcium overload inhibitor BAPTA-AM rescued MeHg-induced neurodevelopmental damage, attenuated the onset of calcium overload, reduced mitochondrial kinetic disturbances and apoptosis. Meanwhile, the activation of the CaM/CaMKII/DRP1 signaling pathway induced by calcium overload was inhibited, and the interaction between DRP1 and BAX was attenuated, which alleviated apoptosis to a certain extent. In summary, our study suggests that MeHg-induced calcium overload may induce disturbed mitochondrial dynamics through activation of the CaM/CaMKII/DRP1 signaling pathway, resulting in neuronal apoptosis.

Revealing mercury species-specific transfer and toxicity mechanisms in placental trophoblasts

Environmental mercury (Hg) follows a biogeochemical cycle resulting in a variety of Hg species. Therefore, human exposure to the three Hg species inorganic Hg via crops and air, methyl Hg through fish consumption and ethyl Hg due to the use as antiseptic agent in medical applications is a rising concern. Especially pregnant women and their developing fetus present a vulnerable population. However, little is known about its transfer and toxicity in placental barrier building cells. Here, Hg species-specific transfer and toxicity in placental trophoblasts, which are the main cell type involved in nutrient transfer, were investigated by using the established BeWo b30 in vitro model. The transfer of inorganic Hg was much lower compared to the organic Hg species and all three species were able to perturb barrier integrity. This was accompanied by a less pronounced cytotoxicity of HgCl2 compared to the two organic species. The energy charge value indicated an increase for inorganic Hg and a decrease for organic Hg compounds. Regarding antioxidative defense, inorganic Hg elevated GSSG levels, while organic Hg decreased GSH. Activity of antioxidative defense related enzymes showed a decrease upon Hg species treatment and all three species induced both apoptotic and necrotic cell death.

Metabolic Profiles of Pregnancy with Polycystic Ovary Syndrome: Insights into Maternal-Fetal Metabolic Communication.

PCOS pregnancies are linked to metabolic disorders affecting maternal and fetal outcomes, with maternal metabolites differing from those in normal pregnancies. To investigate the metabolic communication at the maternal-fetal interface in PCOS pregnancies. Placenta and umbilical cord serum were analyzed using gas chromatography-mass spectrometry. In-depth analysis was performed with clinical characteristics. Placenta and umbilical cord serum were analyzed using gas chromatography-mass spectrometry, alongside clinical characteristics. 45 uncomplicated PCOS pregnancies and 50 normal pregnancies. None. The metabolic characteristics at the maternal-fetal interface in PCOS pregnancies and the underlying mechanisms. A total of 79 metabolites in the placenta and 25 in umbilical cord serum showed significant differences between polycystic ovary syndrome (PCOS) and normal pregnancies. The 10 most significant placental metabolites were identified through receiver operating characteristic (ROC) analysis, 9 of which correlated significantly with maternal serum testosterone levels. Lasso regression analysis identified 4 key placental metabolite combinations: gamma-aminobutyric acid, proline, glycine, and isoleucine, achieving an area under the curve (AUC) of 93.24%. In umbilical cord serum, 6 metabolites differed significantly between PCOS and normal pregnancies, with the highest AUC reaching 76.07%, and 5 of these metabolites showed significant correlations with maternal serum testosterone levels. Nine differential metabolites were shared between the placenta and umbilical cord serum, which also shared metabolic pathways, including ABC transporters and aminoacyl-tRNA biosynthesis, potentially influencing maternal-fetal interactions. This study identifies the metabolomic profile and key pathways in maternal-fetal communication during PCOS pregnancies.

Distinct TYRO3 and PROS1 expression levels contribute to preeclampsia pathogenesis.

Preeclampsia (PE) is a severe placental complication occurring after the 20thweek of pregnancy. PE is associated with inflammation and an increased immune reaction against the fetus. TYRO3 and PROS1 suppress inflammation by clearing apoptotic cells. Disruptions in TYRO3/PROS1 signaling may increase the risk of PE. This study investigated the role of TYRO3/PROS1 signaling in the development of PE using healthy placentae (HP) and preeclamptic placentae (PP) of six pregnant women each. Tissue morphology using hematoxylin and eosin (H&E), TYRO3, MERTK, PROS1, and GAS6 mRNA levels using qPCR and localization and expression levels of TYRO3 and PROS1 using immunohistochemical staining (IHC) were evaluated. The study results show that the levels of TYRO3, MERTK, PROS1 and GAS6 mRNA, as well as TYRO3 protein, increased in PE. TYRO3 expression was observed in extravillous trophoblast (EVTs) and syncytiotrophoblast cells (SCTs). PROS1 was observed in HP fetal vessels through IHC while absent in PP. The reduced presence of PROS1 in the cytotrophoblast layer in PE may indicate a compromised blood-placental barrier. The absence of PROS1 in fetal vessels may suggest potential complement activation and thrombosis. TYRO3, MERTK, PROS1 and GAS6 may help balance impaired inflammation, apoptosis, thrombosis, complement activation and the blood-placental barrier in PE.

Alginate-polylysine-alginate (APA) microencapsulated transgenic human amniotic epithelial cells ameliorate fibrosis in hypertrophic scars.

Hypertrophic scar (HS) is a severe skin fibrosis. Transplanting stem cells carrying anti-fibrotic cytokine genes, like interferon-gamma (IFN-γ), is a novel therapeutic strategy. Human amniotic epithelial cells (hAECs) are ideal seed cells and gene vectors. Microencapsulation creates a favorable environment for transplanted cells. This study investigates the effect of alginate-polylysine-alginate (APA)-microencapsulated hAECs modified with IFN-γ on HS fibrosis. hAECs were isolated from human placentas and characterized. The full-length IFN-γ gene was cloned into the pcDNA3.1 vector to create the recombinant plasmid IFN-γ-pcDNA3.1. This plasmid was then transfected into hAECs, resulting in the generation of IFN-γ-modified hAECs (IFN-γ-hAECs). Subsequently, these IFN-γ-hAECs were microencapsulated with APA to produce APA-IFN-γ-hAECs. In vitro, the release of IFN-γ, as well as the cellular and metabolic activities, growth, proliferation, migration, apoptosis, and trans-differentiation were assessed using HS-derived fibroblasts. In vivo, the weight loss of HS xenografts, collagen fiber arrangement, tissue oxidative stress, and inflammatory response were evaluated using a nude mouse model that had been transplanted with human HS tissues. In vitro, APA-IFN-γ-hAECs exhibited significantly sustained and enhanced IFN-γ release, increased cellular vitality, and inhibited fibroblast growth, proliferation, migration, and trans-differentiation into myofibroblasts. APA-IFN-γ-hAECs also remarkably downregulated extracellular matrix (ECM) components and promoted apoptosis. In vivo, they significantly accelerated the weight reduction of HS xenografts, improved collagen fiber arrangement, and mitigated oxidative stress and inflammation. This study suggests that APA-microencapsulated IFN-γ-hAECs may have potential in alleviating HS fibrosis, offering a new direction for exploring effective clinical HS management strategies.

Paracetamol, its metabolites, and their transfer between maternal circulation and fetal brain in mono- and combination therapies.

Due to its availability and perceived safety, paracetamol is recommended even during pregnancy and for neonates. It is used frequently alone or in combination with other drugs required for the treatment of various chronic conditions. The aim of this study was to investigate potential effects of drug interactions on paracetamol metabolism and its placental transfer and entry into the developing brain. Sprague Dawley rats at postnatal day P4, pregnant embryonic day E19 dams, and non-pregnant adult females were administered paracetamol (15mg/kg) either as monotherapy or in combination with one of seven other drugs: cimetidine, digoxin, fluvoxamine, lamotrigine, lithium, olanzapine, valproate. Concentrations of parent paracetamol and its metabolites (paracetamol-glucuronide, paracetamol-glutathione, and paracetamol-sulfate) in plasma, cerebrospinal fluid (CSF) and brain were measured by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) and their entry into the brain, CSF and transfer across the placenta were estimated. In monotherapy, concentration of parent paracetamol in plasma, CSF, and brain remained similar and at all ages brain entry was unrestricted. In combination therapies, CSF entry of paracetamol increased following co-treatment with olanzapine. Placental transfer of parent paracetamol remained unchanged, however, transfer of paracetamol-sulfate increased with lamotrigine co-administration. Acutely administered paracetamol was more extensively metabolized in adults compared to younger ages resulting in increased concentration of its metabolites with age. Developmental changes in the apparent brain and CSF entry of paracetamol appear to be determined more by its metabolism, rather than by cellular control of its transfer across brain and placental barriers.

Hydrogen sulfide sustains mitochondria functions via targeting mitochondria fission regulator 1 like protein to restore human cytotrophoblast invasion and migration.

Hydrogen sulfide (H2S) is bioactive in mammals. Reduced H2S was observe in pregnancy complications, pre-eclampsia (PE). Our previous data demonstrated that low dose of H2S enhanced cytotrophoblast (CTB) invasion and migration via mitochondria dynamics without knowing the mechanisms. This study was designed to explore the functional regulation of CTB by mitochondrial fission regulator 1 like (MTFR1L) and the mechanisms. By studying human placenta samples and HTR-8/SVneo cell line, MTFR1L was found expressed in CTB. While MTFR1L expression was lower in PE placenta and CTB comparing with Normal pregnancy. Knockdown of MTFR1L decreased CTB invasion and migration, as well as the ATP production, while increased the mitochondria fragmentation, ROS production and mitochondria membrane potential indicating MTFR1L was key regulator of mitochondria. The posttranslational modulation analysis showed enhanced persulfidation of MTFR1L on cystine 222 and 230 by H2S. Mutations of MTFR1LC222/C230 suppressed ATP production, CTB invasion, migration, and increased mitochondria fragmentation, ROS production and mitochondria membrane potential. The present study showed the functional MTFR1L received endogenous CBS/H2S regulation. MTFR1LC222/230 persulfidation by H2S maintained mitochondria morphology and functions thus restored CTB invasion and migration. These findings established a new regulatory pathway for CTB invasion and migration, and provided new targets for PE treatment.

The Impact of Inflammatory Cytokines on Recurrent Pregnancy Loss: A Preliminary Investigation.

Recurrent pregnancy loss (RPL), defined as two or more consecutive miscarriages before 20weeks of gestation, affects 1-2% of couples worldwide. Pro-inflammatory cytokines, such as TNF-α, IL-1β and IL-6 play critical roles in early pregnancy, while anti-inflammatory cytokines like TGF-β and IL-10 promote immune tolerance to prevent harmful inflammatory responses that play important role in placental and fetal development. This aim of the study is to analyse the levels of inflammatory cytokines in blood serum from RPL patients and healthy women (control). The measured cytokines included TNF-α, IL-6, IL-10, TGF-β, CRP, ferritin, IL-1β and IL-4, IFN-γ and IL-17. Using an unpaired t-test and Pearson correlation, significant difference observed between the groups. The results had significantly elevated CRP levels with decreased levels of TGF-β and ferritin (p < 0.05), whereas, IL-1β and IL-4 also found raised indicating a link between systemic inflammation and recurrent miscarriages. IL-4 and CRP increase further suggest potential oxidative stress role in RPL cases. However, no significant differences observed in IL-10, IL-6, or TNF-α level between the groups. This study highlights immune dysregulation as possible contributors to early pregnancy loss, with significant increases in CRP, IL-1β, and IL-4 levels indicating an imbalanced immune response at the maternal-fetal interface. These cytokine elevations may disrupt immune tolerance, suggesting the need for further exploration into cytokine interactions in pregnancy and their potential as an investigatory biomarker and therapeutic target in RPL.