Fetal Cord Serum Research Articles

Expression of sirtuin 2 and 7 in placenta accreta spectrum.

This study aimed to investigate the expression levels of sirtuin 2 and sirtuin 7 in the placenta accreta spectrum to reveal their role in its pathogenesis. A total of 30 placenta accreta spectrum, 20 placenta previa, and 30 controls were experienced. The sirtuin 2 and sirtuin 7 expression levels in the placentas of these groups were determined by Western blot. sirtuin 2 and sirtuin 7 serum levels in the maternal and fetal cord blood were examined by enzyme-linked immunosorbent assay. It was found that sirtuin 7 in placenta accreta spectrum was significantly lower in the placenta compared to the control and placenta previa groups (p<0.05). However, a significant difference was not observed between the sirtuin 2 and sirtuin 7 levels in the maternal and fetal cord serum samples of those three groups (p>0.05). Sirtuin 7 may play an important role in the formation of placenta accreta spectrum. The effect of decreased expression of sirtuin 7 might be tissue-dependent in the placenta accreta spectrum and needs to be investigated further.

2129. Maternal Transmission of SARS-CoV-2-specific Antibodies, but not Cytokines/Chemokines to Neonates Following Infection and Vaccination During Pregnancy

Abstract Background Despite extensive studies of human immune responses to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and coronavirus disease 2019 (COVID-19) vaccination, research examining protective correlates of vertical transmission following maternal exposure in pregnancy remain limited. Here, we characterized antibody and cytokine responses in maternal and cord blood following infection or vaccination at various timepoints during gestation. Methods Spike S1 protein-specific binding antibodies and antibodies capable of blocking the interaction between the receptor binding domain (RBD) and the angiotensin converting enzyme 2 (ACE2) were measured in maternal and cord blood by ELISA. Serum concentrations of 74 cytokines/chemokines were measured by multiplex assay. Humoral responses and cytokine levels from matched maternal and fetal cord sera were compared and examined for potential correlations. Results We observed a highly significant correlation between Spike S1-specific antibody titer and RBD-ACE2 blocking antibody activity between maternal and fetal cord serum (p &amp;lt; 2.2e-16, R &amp;gt; 0.90). Blocking antibody activity was significantly higher for mothers infected during the 3rd trimester compared to earlier trimesters; however, vaccinated mothers developed and transferred higher antibody titers with greater RBD-ACE2 blocking antibody activity to their neonates than infected mothers. Furthermore, vaccine-induced Spike S1 IgG transfer ratios (fetal cord/maternal) were significantly higher than those induced by infection (p = 0.002). Multiplex assay showed significantly elevated levels of 33 cytokines/chemokines, mainly pro-inflammatory in infected maternal serum samples, while the paired fetal cord samples exhibited an anti-inflammatory cytokine predominance. Conclusion Our data support selective vertical transmission of potentially protective humoral responses against SARS-CoV-2, especially following vaccination in the 3rd trimester. The anti-inflammatory cytokine predominance in cord blood that persists despite maternal SARS-CoV-2 infection may offset the adverse outcomes of inflammation in pregnancy for the neonate. Disclosures All Authors: No reported disclosures.

Fetal macrosomia, fetal insulin, and insulin-like growth factor- 1 among neonates in Lagos, Nigeria: A case-control study.

Fetal macrosomia is associated with perinatal injuries. The purpose of this study was to assess the relationship between fetal insulin, insulin-like Growth factor-1(IGF-1), and macrosomia in a resource-limited setting. This was a case-control study at tertiary and secondary health facilities in Lagos, Nigeria. One hundred and fifty mother-neonate pairs were recruited, and their socio-demographic and obstetric history was recorded. Fetal cord venous blood was collected at birth, and neonatal anthropometry was measured within 24hrs of life. Insulin and IGF-1 assay were measured with Enzyme-Linked Immunosorbent Assay (ELISA). Pearson's Chi-square was used to assess the association between categorical variables and macrosomia. Spearman's rank correlation of insulin, IGF-1, and fetal anthropometry was performed. Multivariable logistic regression was used to evaluate the association of insulin and IGF-1 with fetal birth weight. A statistically significant level was set at P-value < 0.05. Macrosomic neonates had mean fetal weight, fetal length, and occipitofrontal circumference (OFC) of 4.15±0.26kg, 50.85±2.09cm and 36.35± 1.22cm respectively. The median Insulin (P = 0.023) and IGF-1 (P < 0.0001) were significantly higher among macrosomic neonates as compared to normal weight babies. Maternal BMI at birth (p = 0.003), neonate's gender (p < 0.001), fetal cord serum IGF-1 (p < 0.001) and insulin assay (P-value = 0.027) were significant predictors of fetal macrosomia. There was positive correlation between cord blood IGF-1 and birth weight (r = 0.47, P-value < 0.001), fetal length (r = 0.30, P-value = 0.0002) and OFC (r = 0.37, P-value < 0.001). Among participating mother-neonate dyad, maternal BMI at birth, neonate's gender, and fetal cord serum IGF-1 and serum insulin are significantly associated with fetal macrosomia.

Fetal and maternal inflammatory response in the setting of maternal intrapartum fever with and without clinical and histologic chorioamnionitis

Both infectious and noninfectious causes of maternal fever have been linked to adverse neonatal outcomes including low Apg0ar scores, respiratory distress, hypotonia, and neonatal seizures. Even in the absence of infection, the occurrence of intrapartum fever is a strong risk factor for poor long-term neonatal developmental outcomes, including encephalopathy, cerebral palsy, and neonatal death. The primary objective of this study was to compare intrapartum and postpartum maternal and fetal umbilical cord serum levels of cytokines RANTES, interferon-ɣ, interleukin-1β, interleukin-2, interleukin-4, interleukin-6, interleukin-8, interleukin-10, interleukin-13, and tumor necrosis factor-α among nonfebrile patients, febrile patients without clinical chorioamnionitis, and febrile patient with clinical chorioamnionitis. This study was conducted at the Richmond University Medical Center from May 15, 2020 to July 16, 2019. During this time, we recruited 30 nonfebrile patients at >36 gestational weeks who were in labor and collected umbilical cord and pre- and postdelivery maternal serum samples to evaluate the cytokine levels. Placentas were collected for pathologic review and to evaluate the histopathologic findings. These results were compared with 121 patients who developed a fever of >38°C during labor. The febrile patients were further divided based on the presence or absence of clinical chorioamnionitis. A secondary analysis was performed based on the presence of absence of histologic chorioamnionitis. Statistical analysis was performed using IBM Statistical Package for the Social Sciences version 25.0. For the 3 group comparisons, a P value of <.017 was considered statistically significant after application of a Bonferroni correction. A total of 151 patients were included in the study; 30 were nonfebrile patients, 46 were febrile patients with a diagnosis of clinical chorioamnionitis, and 75 were febrile patients without clinical chorioamnionitis. Compared with nonfebrile patients, umbilical cord serum interferon-ɣ, interleukin-1β, interleukin-6, interleukin-8, RANTES, and tumor necrosis factor-α levels were elevated in the presence of maternal hyperthermia irrespective of the diagnosis of clinical chorioamnionitis. Interleukin-6 umbilical cord levels were more than doubled from 63.60 pg/mL (6.09-1769.03 pg/mL) in febrile patients with no clinical chorioamnionitis to 135.77 pg/mL (1.86-6004.78 pg/mL) in febrile patients with clinical chorioamnionitis, making it the only cytokine that was significantly different between these 2 groups. When comparing the intrapartum maternal serum, we found a significant elevation in the interleukin-10, RANTES, and tumor necrosis factor-α levels in the febrile group irrespective of the presence of clinical chorioamnionitis when compared with the nonfebrile group. In the postpartum maternal blood evaluations, tumor necrosis factor-α was the only cytokine that was significantly higher in febrile patients than in nonfebrile controls. In the setting of intrapartum fever, maternal cytokine profiles were similar irrespective of the diagnosis of clinical chorioamnionitis. Even in the absence of clinical or histologic chorioamnionitis, maternal hyperthermia induced elevations in fetal cytokines.

Is There A Relationship Between The Umbilical Cord Coiling Index And Oxidative Stress Markers And SGA Fetuses?

Objective: We aimed to investigate the relationship between umblical coiling index and thiol/disulfide balance in SGA newborns to compare with AGA newborns.&#x0D; Material and Methods: Fetal umbilical cord serum samples were collected during labour and the thiol/disulfide homeostasis was measured. After birth, umbilical cords were collected and cord parameters were examined including, length, number of coils, umbilical cord index.&#x0D; Results: Significant decrease in native thiol, total thiol, and a significant increase in disulfide, disulfide/native thiol ratio and disulfide/total thiol ratios were observed in the study group while native thiol/total thiol ratio levels of the two groups did not reach to a statistical significance.&#x0D; Conclusion: The results of this study showed that SGA babies had altered thiol homeostasis in favor of oxidative stress.

Altered cord serum 25-hydroxyvitamin D signaling and placental inflammation is associated with pre-term birth.

Vitamin D is well-known for having anti-inflammatory and immunomodulatory properties. Impaired maternal vitamin D status has been known to increase the risk of adverse pregnancy outcomes like pre-term birth. The present study aims to evaluate the impact of fetal cord serum 25-hydroxyvitamin D-mediated signaling in mediating inflammatory responses in placenta during pre-term birth. For the above purpose, cord serum 25 hydroxyvitamin D 25(OH)D were measured in term (n=20) and pre-term (n=20) born babies using ELISA. Vitamin D downstream signaling has also been checked in placenta (VDR, CYP27B1, cathelicidin LL37) along with expression of inflammatory markers (S100A8, HMGB1, TLR2, p-NF-kappaB) using Western blotting and immunohistochemistry. Pearson correlation model was used to do correlation study. Compared with term born babies (59.31±3.476), decline in cord serum 25(OH)D levels is observed in pre-term born babies (22.26±1.083, P=<0.0001) that showed strong positive correlation with gestational age (r=.9368***) and birthweight (r=.9559***). On the other hand, vitamin D signaling markers were found to be downregulated and inflammatory markers were upregulated in placental tissue of pre-term born babies. Thus, our study demonstrated that insufficient cord 25(OH)D levels may disturb the homeostasis of inflammation in placenta. Altered cord serum 25(OH)D mediated anti-inflammatory signaling may be acting as trigger signals in modulating inflammatory responses in placenta and eliciting premature activation of spontaneous labor in pre-term birth.

High maternal selenium levels are associated with increased risk of congenital heart defects in the offspring.

To examine whether maternal hair or cord blood selenium levels are associated with infant congenital heart defects (CHDs). A hospital-based case-control study was performed with samples collected from 2010 to 2014. Selenium levels in maternal hair and fetal cord serum were measured using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Data from medical records of infants with CHDs were collected. Logistic regression analysis was employed to examine the independent association between maternal selenium exposure and fetal CHD. In total, 888 pregnant women were included in this study. Median (interquartile range) selenium levels in of maternal hair was similar in the control and CHD group. A significant association was detected between increased maternal hair selenium concentration and CHDs in offspring (adjusted odds ratio [aOR] 3.57, 95 %CI, 1.90-6.70). The association was present in all CHD groups, including septal defects, conotruncal defects, left ventricular outflow tract obstruction, right ventricular outflow tract obstruction, and anomalous pulmonary venous return. High maternal selenium level is significantly associated with the occurrence of fetal CHD.

Role of vitamin D in cell-cell interaction of fetal endothelial progenitor cells and umbilical cord endothelial cells in a preeclampsia-like model.

Maternal endothelial dysfunction is a cental feature of preeclampsia (PE), a hypertensive disorder of pregnancy. Factors in the maternal circulation are thought to contribute to this endothelial dysfunction. Although understudied, factors in the fetal circulation may influence fetal endothelial cell interactions with endothelial progenitor cells as critical steps in placental angiogenesis. We hypothesize that cell-cell interactions that are important for pregnancy health are impaired by fetal serum from PE pregnancies and that 1,25(OH)2-vitamin D3 attenuates the negative effects of this serum on cell function. We tested the ability of fetal cord blood-derived endothelial progenitor cells [endothelial colony-forming cells (ECFCs)] to invade into established monolayers and capillary tubule-like structures of human fetal umbilical venous endothelial cells (HUVECs), while in the presence/absence of fetal cord serum from uncomplicated or PE pregnancies, and tested the ability of 1,25(OH)2-vitamin D3 to modulate the serum-mediated effects. PE cord serum reduced the invasion of fetal ECFCs into HUVEC monolayers or tubule networks. Vitamin D attenuated these effects of PE fetal serum on endothelial functional properties. Immunocytochemical studies revealed involvement of VE-cadherin contacts in interactions between ECFCs and mature fetal endothelial cells. PE cord serum reduces the ability of fetal endothelial progenitor cells to incorporate into fetal endothelial cell networks. Physiologic concentrations of vitamin D reverse these PE serum-mediated effects. These data appear consistent with lines of evidence that vitamin D has antipreeclampsia effects.

Evaluation of fetal serum thiol/disulphide homeostasis in deliveries complicated by nuchal cord

Aim: To investigate the serum thiol/disulphide homeostasis in deliveries complicated by nuchal cord (NC) and to compare the results with healthy deliveries (without NC).Methods: This prospective controlled study included 48 pregnant women complicated by NC and 48 similar gestational aged healthy pregnant women during labor. Fetal umbilical cord serum samples were collected during labor and the thiol/disulphide homeostasis was measured by using an automated assay method. The patients were followed up until end of the delivery and perinatal outcomes were recorded.Results: Fetal umbilical cord native thiol, total thiol, and disulphide levels as well as disulphide/native thiol and disulphide/total thiol ratios are impaired in labor with the presence of NC. There were no statistically significant differences in terms of maternal and gestational age at delivery and maternal number of gravida and parity, fetal gender, fifth Apgar scores <7, mode of delivery and fetal birth weight between groups. The group of patients with NC had higher emergency C/S numbers indicated for fetal distress and lower first Apgar scores <7. There were no neonatal intensive care unit admissions among these babies.Conclusions: Maternal serum thiol/disulphide homeostasis reflect transient effects of NC during labor regardless of labor type. Vaginal delivery can be safely and successfully performed in pregnancies complicated with NC.

Fetal cord serum prolactin (PRL) levels in normal and abnormal pregnancies and its relation to development of RDS

Fetal cord serum prolactin (PRL) levels in normal and abnormal pregnancies and its relation to development of RDS

339: Maternal and fetal umbilical cord levels of chemokines in normal and PreE pregnancies - A prospective trial

Preeclampsia (PreE) is the de novo development of hypertension and proteinuria after 20 weeks of gestation affecting 3-8% of pregnancies, and has a link to alterations of feto-placental stress that pass to the offspring causing detrimental effects. We aimed at comparing the clinical outcomes and chemokines from the maternal and cord blood between patients with and without PreE. We recruited 20 normal pregnant (NP) and PreE patients in an IRB approved prospective cohort at our institution. We evaluated maternal age, body mass index (BMI), blood pressure (BP), and gestational age (GA) at delivery along with measured serum levels of six chemokines in maternal-fetal paired samples using ELISA test. The markers are; Human CCL4/MIP-1beta, Human CXCL10/IP-10, Eotaxin/CCL11, RANTES (CCL5), Human MCP1, Human M-CSF. Maternal: The systolic blood pressure (SBP) and diastolic blood pressure (DBP) were higher in PreE (SBP 162 ± 17; DBP 97 ± 12) compared to NP (SBP 121 ± 3; DBP 75 ± 9; p <0.05). GA at delivery in weeks was lower in PreE (35.1 ± 3.8) compared to NP (39.2 ± 0.3; p <0.05), as well as birth weight in PrE (2556 ± 977) compared to NP (3385 ± 328; p <0.011) indicating a link to intrauterine growth restriction (IUGR). We did not find a difference in maternal BMI between the two groups. Biomarkers: For all six chemokines CCL4, CXCL10, CCL11, CCL5, MCP1, and M-CSF, maternal and fetal cord serum chemokines levels differ between subjects with and without PreE (p < 0.001). There is also a difference between the maternal and fetal levels for the six chemokines (<0.01). The fetal levels were compared to associated maternal levels for each of the six chemokines using t test and Mann-Whitney U test. Women with PreE had higher BP, preterm deliveries, and fetal IUGR. However, in this cohort the women were of similar age, size, and parity prior to the onset of pregnancy. The six chemokines in the samples from maternal and fetal circulation are greater in the PreE group, and each of the six chemokines samples from the fetuses had values greater than maternal samples which could implicate placental cells as the source. PreE alters the intrauterine environment and activates the detrimental signaling that is transferred to the fetus resulting in the aforementioned complications.

The Tissue Culture Laboratory of Dr. George Otto Gey 60yrs ago as recalled by a former student.

George Otto Gey was a pioneer in tissue culture, having introduced the roller drum, the HeLa cell line, and the use of human fetal cord serum and beef embryo extract. During his career (1920s-1960s), the field of tissue culture was in its infancy and not yet dependent upon commercial biological supply houses. While the early techniques of cell culture have been greatly improved upon, of historical interest may be personal observations of the Gey Tissue Culture Laboratory, Johns Hopkins Medical School, as recalled by a medical student working there in the 1950s. Dr. Gey served as a founding member and executive of the Tissue Culture Commission (TCC) and became the first president of the Tissue Culture Association (TCA).

Peroxiredoxins prevent oxidative stress during human sperm capacitation.

Do peroxiredoxins (PRDXs) control reactive oxygen species (ROS) levels during human sperm capacitation? PRDXs are necessary to control the levels of ROS generated during capacitation allowing spermatozoa to achieve fertilizing ability. Sperm capacitation is an oxidative event that requires low and controlled amounts of ROS to trigger phosphorylation events. PRDXs are antioxidant enzymes that not only act as scavengers but also control ROS action in somatic cells. Spermatozoa from infertile men have lower levels of PRDXs (particularly of PRDX6), which are thiol-oxidized and therefore inactive. Semen samples were obtained from a cohort of 20 healthy nonsmoker volunteers aged 22-30 years old over a period of 1 year. Sperm from healthy donors was capacitated with fetal cord serum ultrafiltrate (FCSu) in the absence or presence of thiostrepton (TSP), inhibitor of 2-Cys PRDXs or 1-Hexadecyl-3-(trifluoroethyl)-sn-glycero-2-phosphomethanol lithium (MJ33), inhibitor of calcium independent-phospholipase A2 (Ca2+-iPLA2) activity of PRDX6, added at different times of incubation. Capacitation was also induced by the dibutyryl cAMP+3-isobuty1-1-methylxanthine system. Sperm viability and motility were determined by the hypo-osmotic swelling test and computer-assisted semen analysis system, respectively. Capacitation was determined by the ability of spermatozoa to undergo the acrosome reaction triggered by lysophosphatidylcholine. Percentages of acrosome reaction were obtained using the FITC-conjugated Pisum sativum agglutinin assay. Phosphorylation of tyrosine residues and of protein kinase A (PKA) substrates were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis immunoblotting with specific antibodies. Actin polymerization was determined by phalloidin labeling. TSP and MJ33 prevented sperm capacitation and its associated actin polymerization in spermatozoa incubated with 10% FCSu (capacitation inducer) compared to non-capacitated controls (P < 0.05) without altering sperm viability. PKA substrates and tyrosine phosphorylations were prevented in FCSu-treated spermatozoa in a differential fashion depending on the type and the time of addition of the inhibitor used compared to non-capacitated controls (P < 0.05). TSP and MJ33 promoted an increase of lipid peroxidation in spermatozoa (P < 0.01) and these levels were higher in those spermatozoa incubated with the inhibitors and FCSu compared to those capacitated spermatozoa incubated without the inhibitors (P < 0.0001). Inhibition of 2-Cys PRDXs by TSP generated an oxidative stress in spermatozoa, affecting their viability compared to controls (P < 0.05). This oxidative stress was prevented by nuclephile D-penicillamine (PEN). MJ33 also promoted an increase of lipid peroxidation and impaired sperm viability compared to non-treated controls (P < 0.05) but its effect was not circumvented by PEN, suggesting that not only peroxidase but also Ca2+-iPLA2 activity of PRDX6 are necessary to guarantee viability in human spermatozoa. Not applicable. We focused on the global effect of PRDXs inhibitors on human sperm capacitation and in two of its associated phosphorylation events. Thus, other phosphorylation events and mechanisms necessary for capacitation may also be affected. PRDXs are the major antioxidant system in ejaculated spermatozoa and are necessary to allow spermatozoon to achieve fertilizing ability (capacitation and acrosome reaction). This research was supported by Canadian Institutes of Health Research (MOP 133661) and the Fonds de Recherché en Santé Quebec (FRSQS #22151) to C.O. The authors have nothing to disclose.

Antenatal factors modulate hearing screen failure risk in preterm infants

ObjectiveThe objective of this study was to characterise the effects of antenatal inflammatory factors and maternal therapies on neonatal hearing screen outcomes in very low birthweight infants.MethodsWe conducted a retrospective...

Role of Fetal Serum Prolactin in Pregnancy

&lt;p&gt;In our study we have tried to to study the comparison of fetal cord serum prolactin level in normal and abnormal pregnancies and correlate the cord serum prolactin level with occurrence of HMD and to establish the fact that If cord serum prolactin is less, then there might be the chances of neonate developing HMD .study was conducted in Department of Obstetrics and Gynaecology, Datta Meghe Institute Of Medical Sciences, Sawangi (Meghe), Wardha (1st September 2005 to 31st August 2007: on 160 randomly selected booked and unbooked women GROUP A 80 women with normal pregnancies GROUP B 80 women with abnormal pregnancies.&lt;/p&gt;&lt;p&gt;&lt;br /&gt;In a prospective study umbilical cord serum prolactin level was found to be lower in women with abnormal pregnancy.&lt;/p&gt;

Neonatal Genetic Variation in Steroid Metabolism and Key Respiratory Function Genes and Perinatal Outcomes in Single and Multiple Courses of Corticosteroids.

The aim of the study is to evaluate the association of steroid metabolism and respiratory gene polymorphisms in neonates exposed to antenatal corticosteroids (ACS) with respiratory outcomes, small for gestational age (SGA), and response to repeat ACS. This candidate gene study is a secondary analysis of women enrolled in a randomized controlled trial of single versus weekly courses of ACS. Nineteen single nucleotide polymorphisms (SNPs) in 13 steroid metabolism and respiratory function genes were evaluated. DNA was extracted from placenta or fetal cord serum and analyzed with TaqMan genotyping. Each SNP was evaluated for association via logistic regression with respiratory distress syndrome (RDS), continuous positive airway pressure (CPAP)/ventilator use (CPV), and SGA. CRHBP, CRH, and CRHR1 minor alleles were associated with an increased risk of SGA. HSD11B1 and SCNN1B minor alleles were associated with an increased likelihood of RDS. Carriage of minor alleles in SerpinA6 was associated with an increased risk of CPV. CRH and CRHR1 minor alleles were associated with a decreased likelihood of CPV. Steroid metabolism and respiratory gene SNPs are associated with respiratory outcomes and SGA in patients exposed to ACS. Risks for respiratory outcomes are affected by minor allele carriage as well as by treatment with multiple ACS.

BPA and BPA metabolites in umbilical cord serum during mid-gestation in a northern California cohort

Background: Bisphenol A (BPA) is a chemical widely used in numerous consumer products, resulting in nearly universal exposure among the U.S. population. BPA is an endocrine modulator and prenatal exposure is associated with numerous reproductive and developmental effects in animals. However, little is known about human fetal exposure or the ability of the fetus to metabolize BPA during mid-gestation. Objective: To measure fetal exposure to BPA and its metabolites during the 2nd trimester of pregnancy. Methods: We used targeted liquid chromatography-tandem mass spectrometry (LC/MS-MS) to measure unconjugated BPA and the two predominant BPA metabolic conjugates-BPA glucuronide and BPA sulfate- in fetal umbilical cord serum and maternal urine collected from voluntary 2nd trimester pregnancy terminations. Results: We detected unconjugated BPA (GM 0.16 ng/ml; range &lt;LOD-52.26 ng/ml), BPA glucuronide (0.14 ng/mL; range &lt;LOD-5.41 ng/mL) and BPA sulfate (GM 0.32 ng/mL; range &lt;LOD-12.65 ng/mL) in all 85 cord serum samples examined. The conjugated to unconjugated ratio ranged from &lt; 0.01 to greater than 300. Conjugated levels exceeded unconjugated levels in about three-quarters of the samples, but unconjugated levels were higher in samples with total BPA above the GM. In a subset of 29 matched maternal urine samples we found positive correlations between BPA conjugates in urine and cord blood (Spearman rho correlation 0.38 and 0.34 for BPA sulfate and BPA glucuronide respectively (p-value 0.07 and 0.04)). Total BPA levels were not correlated. Conclusion: We found universal fetal exposure to BPA with the novel finding of detectable BPA sulfate during mid-gestation among our study cohort. A small subset of fetuses had unconjugated BPA levels that exceeded what was measured in paired maternal urine samples.

Human Sperm Chromatin Undergoes Physiological Remodeling During In Vitro Capacitation and Acrosome Reaction

Capacitation (CAP) and acrosome reaction (AR) are sequential processes of sperm activation. Beside the known ionic, membrane, and transduction events and final release of proteolytic enzymes that help sperm movement toward the egg, chromatin changes, such as a physiological remodeling, are also possible. Our aims were to ascertain that CAP and AR do not induce DNA damage and to evaluate changes occurring in the human sperm head during these physiological processes using cytochemical stains. Percollpurified spermatozoa from normal donors were incubated in Biggers, Whitten, and Whittingham medium ± fetal cord serum ultrafiltrate (CAP inducer) and then with lysophosphatidylcholine (AR inducer). CAP and AR were associated with increases in aniline blue (AB, for histones; ∼70%) and toluidine blue (TB, for chromatin compaction; ∼40%) staining but had no influence on that of chromomycin A3 (for protamines). The increase (∼40%) in iodoacetamide-fluorescein (IAF, for sulfhydryl groups) staining observed during CAP was absent after AR. CAP and AR did not damage DNA (percentage of DNA fragmentation index remained low) nor affect histone content. CAP, and even more AR, primed sperm heads to decondense (∼80% and ∼140% increases, respectively) when challenged with sodium dodecyl sulfate + dithiothreitol. Interestingly, induced decondensation correlated with all other tests (CAP, AB, TB, and IAF). Therefore, the data strongly support a physiological remodeling of nondamaged human sperm chromatin during CAP and AR, and modifications are probably interlinked and help prepare chromatin for postfertilization events.

2098 REMODELING OF HUMAN SPERM CHROMATIN DURING IN VITRO CAPACITATION AND ACROSOME REACTION: EVIDENCE FROM CYTOCHEMICAL TESTS AND CORRELATIONS BETWEEN ASSAYS

You have accessJournal of UrologyInfertility: Physiology, Pathophysiology, Basic Research1 Apr 20122098 REMODELING OF HUMAN SPERM CHROMATIN DURING IN VITRO CAPACITATION AND ACROSOME REACTION: EVIDENCE FROM CYTOCHEMICAL TESTS AND CORRELATIONS BETWEEN ASSAYS Eve de Lamirande, Maria San Gabriel, Naif Alhathal, and Armand Zini Eve de LamirandeEve de Lamirande Montreal, Canada More articles by this author , Maria San GabrielMaria San Gabriel Montreal, Canada More articles by this author , Naif AlhathalNaif Alhathal Montreal, Canada More articles by this author , and Armand ZiniArmand Zini Montreal, Canada More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.2265AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Capacitation (CAP) and acrosome reaction (AR), are sequential processes of sperm activation. Beside the known ionic, membrane and transduction events and final release of proteolytic enzymes to help sperm movement towards the egg, changes to chromatin are also expected. These should involve remodeling rather than DNA damage. Our aims were evaluate the changes in human sperm head using cytochemical stains and to confirm that CAP/AR do not include DNA damage. METHODS Percoll-washed spermatozoa were incubated in BWW +/− fetal cord serum ultrafiltrate (10%;3.5h;CAP) and, subsequently, with lysophosphatidylcholine (2.5 μM; 30 min, AR). Then, ethanol-fixed spermatozoa were used for: PSA-FITC (for AR), aniline blue (AB, for histones), chromomycin A3 (CMA3, for protamines), toluidine blue (TB, for chromatin compaction), iodoacetamide-fluorescein (IAF, for sulfhydryl groups), induced decondensation, sperm chromatin structure assay (DNA fragmentation index, DFI) and immunoblotting (for histone, H2B). RESULTS CAP/AR was associated with similar increases in staining for AB (∼75%) and TB (∼50%) but had no inflence on CMA3. The increase (∼55%) in IAF staining observed during CAP was abscent after AR. CAP/AR did not damage DNA (DFI remained low) nor affect histone content. CAP, and even more AR, primed sperm heads to decondense (∼100% and ∼160% increases, respectively) when challenged with SDS (1%) + DTT (0.1 mM). Interestingly, induced decondensation correlated with all other tests CAP, AB, TB, IAF; also CAP was related to AB and TB stains. CONCLUSIONS The data strongly support human sperm chromatin remodeling during CAP/AR, these modifications are interlinked and probably help prepare chromatin for after-fertilization events. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e845-e846 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Eve de Lamirande Montreal, Canada More articles by this author Maria San Gabriel Montreal, Canada More articles by this author Naif Alhathal Montreal, Canada More articles by this author Armand Zini Montreal, Canada More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

2099 IMPAIRED CLEARANCE OF APOPTOTIC GERM CELLS CAUSES IMPAIRED SPERMATOGENESIS ASSOCIATED WITH LEYDIG CELL HYPERPLASIA AND INCREASED TESTOSTERONE LEVELS

changes in human sperm head using cytochemical stains and to confirm that CAP/AR do not include DNA damage. METHODS: Percoll-washed spermatozoa were incubated in BWW / fetal cord serum ultrafiltrate (10%;3.5h;CAP) and, subsequently, with lysophosphatidylcholine (2.5 M; 30 min, AR). Then, ethanol-fixed spermatozoa were used for: PSA-FITC (for AR), aniline blue (AB, for histones), chromomycin A3 (CMA3, for protamines), toluidine blue (TB, for chromatin compaction), iodoacetamide-fluorescein (IAF, for sulfhydryl groups), induced decondensation, sperm chromatin structure assay (DNA fragmentation index, DFI) and immunoblotting (for histone, H2B). RESULTS: CAP/AR was associated with similar increases in staining for AB ( 75%) and TB ( 50%) but had no inflence on CMA3. The increase ( 55%) in IAF staining observed during CAP was abscent after AR. CAP/AR did not damage DNA (DFI remained low) nor affect histone content. CAP, and even more AR, primed sperm heads to decondense ( 100% and 160% increases, respectively) when challenged with SDS (1%) DTT (0.1 mM). Interestingly, induced decondensation correlated with all other tests CAP, AB, TB, IAF; also CAP was related to AB and TB stains. CONCLUSIONS: The data strongly support human sperm chromatin remodeling during CAP/AR, these modifications are interlinked and probably help prepare chromatin for after-fertilization events.