Neutrophils are often exclusively considered as a first-line innate immune defence, able to rapidly kill or trap pathogens and causing in case of over-activation tissue damage. In the female reproductive tract, however, the presence and activity of neutrophils seems to be tightly regulated. Major players in orchestrating this regulation are cyclical steroid sex hormones present during the menstrual cycle and pregnancy. This review describes the role of sex hormones in regulating directly or indirectly the functionality of neutrophils, the role of neutrophils during fertilization and pregnancy and in controlling viral, fungal and bacterial infection. This review also discusses the consequence of overt neutrophil activation in pregnancy pathologies.

IntroductionThe dysregulation of maternal-fetal immune tolerance is one of the proposed mechanisms leading to preeclampsia. Galectins are key regulator proteins of the immune response in vertebrates and maternal-fetal immune tolerance in eutherian mammals. Previously we found that three genes in a Chr19 cluster encoding for human placental galectin-13 (PP13), galectin-14 and galectin-16 emerged during primate evolution and may confer immune tolerance to the semi-allogeneic fetus.Materials and MethodsThis study involved various methodologies for gene and protein expression profiling, genomic DNA methylation analyses, functional assays on differentiating trophoblasts including gene silencing, luciferase reporter and methylation assays. These methods were applied on placental specimens, umbilical cord blood cells, primary trophoblasts and BeWo cells. Genomic DNA sequences were analyzed for transposable elements, transcription factor binding sites and evolutionary conservation.Results and DiscussionThe villous trophoblastic expression of Chr19 cluster galectin genes is developmentally regulated by DNA methylation and induced by key transcription factors of villous placental development during trophoblast fusion and differentiation. This latter mechanism arose via the co-option of binding sites for these transcription factors through promoter evolution and the insertion of an anthropoid-specific L1PREC2 transposable element into the 5’ untranslated region of an ancestral gene followed by gene duplication events. Among placental Chr19 cluster galectin genes, the expression of LGALS13 and LGALS14 is down-regulated in preterm severe preeclampsia associated with SGA. We reveal that this phenomenon is partly originated from the dysregulated expression of key transcription factors controlling trophoblastic functions and galectin gene expression. In addition, the differential DNA methylation of these genes was also observed in preterm preeclampsia irrespective of SGA.ConclusionsThese findings reveal the evolutionary origins of the placental expression of Chr19 cluster galectins. The complex dysregulation of these genes in preeclampsia may alter immune tolerance mechanisms at the maternal-fetal interface.

Workshops are an important part of the IFPA annual meeting as they allow for discussion of specialized topics. At IFPA meeting 2012 there were twelve themed workshops, four of which are summarized in this report. These workshops related to various aspects of placental biology: 1) epigenetics and imprinting in the placenta; 2) growth factors and villous trophoblast differentiation; 3) role of the placenta in regulating fetal exposure to xenobiotics during pregnancy; 4) infection and the placenta.

Introduction: We found isolated or clustered trophoblasts in the chorionic connective tissue of the extraplacental membranes, and defined this novel histologic feature as the "trophoblast islands of the chorionic connective tissue" (TICCT). This study was conducted to determine the clinical significance of TICCT.Methods: Immunohistochemistry for cytokeratin-7 was performed on the chorioamniotic membranes (N = 2155) obtained from singleton pregnancies of 1199 uncomplicated term and 956 preterm deliveries. The study groups comprised 1236 African American and 919 Hispanic women. Gestational age ranged from 24(+0) weeks to 41(+6) weeks. Multiple logistic regression analysis was performed to investigate the magnitude of association between patient characteristics and the presence of TICCT.Results: The likelihood of TICCT was significantly associated with advancing gestational age both in term (OR: 1.29,95% CI: 1.16-1.45, p < 0.001) and preterm deliveries (OR: 1.19, 95% CI: 1.07-1.32, p = 0.001). Hispanic women were less likely than African American women to have TICCT across gestation in term (OR: 0.23, 95% CI: 0.18-031, p < 0.001) and preterm pregnancies (OR: 0.41, 95% CI: 0.29-0.58, p < 0.001). Women with a female fetus were significantly more likely to have TICCT than women with a male fetus, in both term (OR: 1.64, 95% CI: 128-2.11, p < 0.001) and preterm gestations (OR: 2.04, 95% CI: 1.46-2.85, p < 0.001). TICCT was 40% less frequent in the presence of chronic placental inflammation [term (OR: 0.60, 95% CI: 0.45-0.81, p = 0.001) and preterm gestations (OR: 0.58, 95% CI: 0.40-0.84, p = 0.003)] and in parous women at term (OR: 0.60, 95% CI: 0.44-0.81, p = 0.001).Conclusions: Our findings suggest that the duration of pregnancy, fetal sex, and parity may influence the behavior of extravillous trophoblast and placental mesenchymal cells. Published by Elsevier Ltd.

Background: The involvement of the placenta in the pathogenesis of preeclampsia and HELLP syndrome is well established, and placental lesions are also similar in these two syndromes. Here we aimed to examine the placental transcriptome and to identify candidate biomarkers in early-onset preeclampsia and HELLP syndrome.Methods: Placental specimens were obtained at C-sections from women with early-onset preeclampsia and HELLP syndrome, and from controls who delivered preterm or at term. After histopathological examination, fresh-frozen placental specimens were used for microarray profiling and validation by qRT-PCR. Differential expression was analysed using log linear models while adjusting for gestational age. Gene ontology and pathway analyses were used to interpret gene expression changes. Tissue microarrays were constructed from paraffin-embedded placental specimens and immunostained.Results: Placental gene expression was gestational age-dependent among preterm and term controls. Out of the 350 differentially expressed genes in preeclampsia and 554 genes in HELLP syndrome, 224 genes (including LEP, CGB, LHB, INHA, SIGLEC6, PAPPA2, TREM1, and FLT1) changed in the same direction (elevated or reduced) in both syndromes. Many of these encode proteins that have been implicated as biomarkers for preeclampsia. Enrichment analyses revealed similar biological processes, cellular compartments and biological pathways enriched in early-onset preeclampsia and HELLP syndrome; however, some processes and pathways (e.g., cytokine cytokine receptor interaction) were over-represented only in HELLP syndrome.Conclusion: High-throughput transcriptional and tissue microarray expression profiling revealed that placental transcriptomes of early-onset preeclampsia and HELLP syndrome largely overlap, underlying a potential common cause and pathophysiologic processes in these syndromes. However, gene expression changes may also suggest a more severe placental pathology and pronounced inflammatory response in HELLP syndrome than in preeclampsia. (C) 2010 Elsevier Ltd. All rights reserved.

To examine the relationship between abnormalities in uterine (UtA) and/or umbilical artery (UA) Doppler velocimetry and maternal plasma concentrations of soluble endoglin (sEng) in patients with pre‐eclampsia (PE).

Pre-eclampsia (PE) is thought to be causally related to an “anti-angiogenic state” characterized by increased concentrations of soluble endoglin (Nature Medicine 2006; 12: 642–9) and soluble VEGFR-1 (NEJM 2004; 350 : 672–83). This “anti-angiogenic state” may be reflected by changes in the impendence to flow in the maternal and fetal circulations. The objective of this study was to examine the relationship between abnormalities in uterine (UT) and/or umbilical artery (UA) Doppler velocimetry and maternal plasma concentration of the powerful anti-angiogenic factor, endoglin, in patients with PE. A cross-sectional study was conducted in normal pregnant women (n = 135) and patients with PE (n = 69). Patients with PE were sub-classified into four groups according to the results of UT and UA artery Doppler velocimetry. Plasma concentrations of soluble endoglin (sEng) were determined by ELISA. 1) Patients with an abnormal UT and UA Doppler velocimetry had the highest median plasma concentration of sEng than any other diagnostic group (Kruskal Wallis P < 0.05, see table); 2) Women with PE and normal Doppler velocimetry in both vessels had the lowest median plasma concentration of sEng; 3) Plasma sEng concentrations increased as a function of gestational age (GA) (Spearman's rho = 0.4; P < 0.001). Yet, the patients with the highest median sEng concentration were those with abnormal maternal and fetal Doppler velocimetry and the lowest GA (P < 0.05). Abnormalities of impedance to blood flow in the uterine and umbilical circulation are associated with an excess of sEng in the circulation of mothers with PE. These findings suggest that the severity of the anti-angiogenic state in PE is expressed by abnormalities of Doppler velocimetry.

Utero-placental ischemia has been implicated as one of the mechanisms of disease in SGA. Recently, an anti-angiogenic state (reflected by elevation of soluble endoglin concentrations in maternal blood) has been described in mothers with SGA fetuses without preeclampsia and detected as early as the first trimester of pregnancy. The objective of this study was to determine if mothers of SGA fetuses with and without Doppler abnormalities have different profiles of the anti-angiogenic factor, soluble endoglin (sEng). A cross-sectional study was conducted in 46 pregnancies which led to the birth of SGA neonates. SGA were sub-classified according to the results of uterine (UT) and umbilical (UA) artery Doppler velocimetry. The control group consisted of normal pregnant women who delivered appropriate for gestational age neonates (n = 135). Plasma concentrations of sEng were determined by a sensitive and specific immunoassay. 1) Mothers with SGA fetuses with an abnormal Doppler velocimetry (in the UT or the UA) had a higher median plasma concentration of sEng than those without these abnormalities (both P < 0.05; see Table); 2) Mothers with SGA fetuses and abnormal UA Doppler velocimetry (utero-placental insufficiency) had higher sEng concentrations than both mothers with SGA fetuses with normal Doppler velocimetry and the control group (P < 0.05 for both). An increased impedance to blood flow in the umbilical circulation and in the uterine circulation is strongly associated with an excess availability of the anti-angiogenic factor in mothers with SGA fetuses.

Understanding the mechanisms of disease responsible for the syndrome of preeclampsia as well as early risk assessment is still a major challenge. The concentrations of circulating proteins in maternal blood such as placental growth factor, soluble vascular endothelial growth factor receptor-1 and soluble endoglin are altered weeks before the onset of clinical symptoms of the syndrome. Recently, other proteins in maternal serum, such as activin A, inhibin A, PAPP-A, and PP13 have been suggested to be of value in first trimester risk assessment. Since preeclampsia is a syndrome, it seems unlikely that a single test will predict all forms of preeclampsia. This realization has led to the formulation of a new conceptual framework suggesting that a combination of markers (biochemical and/or biophysical) may be required to conduct comprehensive risk assessment for the syndrome.

The workshop focused on proteins of embryonic, placental or maternal origin involved in unique networks of interactions in establishing and maintaining healthy pregnancy. Special attention was given to various proteins throughout the full-time course of pregnancy, starting from the preimplantation period until labor. Structural and functional data on different polypeptides were shown, intending to evaluate their possible clinical role in healthy or complicated pregnancies and the associated negative consequences. Workshop presentations given by the participants are summarized.

Objectives To determine the clinical significance of the presence of amniotic fluid (AF) 'sludge' among asymptomatic patients at high risk for spontaneous preterm delivery.Methods This retrospective case-control study included 281 patients with (n = 66) or without (n = 215) AF 'sludge', who underwent transvaginal ultrasound examination between 13 and 29 completed weeks of gestation. Patients with threatened preterm labor, multiple gestation, fetal anomalies, placenta previa or uterine contractions were excluded.Results The prevalence of AF 'sludge' in the study population was 23.5% (66/281). The rates of spontaneous preterm delivery at < 28 weeks, < 32 weeks, < 35 weeks and < 37 weeks of gestation were 14.7% (29/197), 21.3% (46/216), 28.7% (62/216) and 42.1% (91/216), respectively. Patients with 'sludge' had: (1) a higher rate of spontaneous preterm delivery at < 28 weeks (46.5% (20/43) vs. 5.8% (9/154); P < 0.001), < 32 weeks (55.6% (25/45) vs. 12.3% (21/171); P < 0.001) and < 35 weeks (62.2% (28/45) vs. 19.9% (34/171); P < 0.001); (2) a higher frequency of clinical chorioamnionitis (15.2% (10/66) vs. 5.1% (11/215); P = 0.007), histologic chorioamnionitis (61.5% (40/65) vs. 28% (54/193); P < 0.001) and funisitis (32.3% (21/65) vs. 19.2% (37/193); P = 0.03); (3) a higher frequency of preterm prelabor rupture of membranes (PROM) (39.4% (26/66) vs. 13.5% (29/215); P < 0.001), lower gestational age at preterm PROM (median 24.7 (interquartile range (IQR), 22.3-28.1) weeks vs. 32.3 (IQR, 27.7-34.8) weeks; P < 0.001); and (4) shorter median ultrasound-to-delivery interval ('sludge' positive 127 days (95% CI, 120-134 days) vs. 'sludge' negative 161 days (95% CI, 153-169 days); P < 0.001) and ultrasound-to-preterm PROM interval ('sludge' positive 23 days (95% CI, 7-39 days) vs. 'sludge' negative 57 days (95% CI, 38-77 days); P = 0.003) than those without 'sludge'. AF 'sludge' was an independent explanatory variable for the occurrence of spontaneous preterm delivery at < 28 weeks, < 32 weeks and < 35 weeks, preterm PROM, microbial invasion of the amniotic cavity (MIAC) and histologic chorioamnionitis. Moreover, the combination of a cervical length < 25 mm and 'sludge' conferred an odds ratio of 14.8 and 9.9 for spontaneous preterm delivery at < 28 weeks and < 32 weeks, respectively.Conclusions AF 'sludge' is an independent risk factor for spontaneous preterm delivery, preterm PROM, MIAC and histologic chorioamnionitis in asymptomatic patients at high risk for spontaneous preterm delivery. Furthermore, the combination of 'sludge' and a short cervix confers a higher risk for spontaneous preterm delivery at < 28 weeks and < 32 weeks than a short cervix alone. Copyright (C) 2007 ISUOG. Published by John Wiley & Sons, Ltd.

High‐dimensional biology (HDB) refers to the simultaneous study of the genetic variants (DNA variation), transcription (messenger RNA [mRNA]), peptides and proteins, and metabolites of an organ, tissue, or an organism in health and disease. The fundamental premise is that the evolutionary complexity of biological systems renders them difficult to comprehensively understand using only a reductionist approach. Such complexity can become tractable with the use of ‘omics’ research. This term refers to the study of entities in aggregate. The current nomenclature of ‘omics’ sciences includes genomics for DNA variants, transcriptomics for mRNA, proteomics for proteins, and metabolomics for intermediate products of metabolism. Another discipline relevant to medicine is pharmacogenomics. The two major advances that have made HDB possible are technological breakthroughs that allow simultaneous examination of thousands of genes, transcripts, and proteins, etc., with high‐throughput techniques and analytical tools to extract information. What is conventionally considered hypothesis‐driven research and discovery‐driven research (through ‘omic’ methodologies) are complementary and synergistic. Here we review data which have been derived from: 1) genomics to examine predisposing factors for preterm birth; 2) transcriptomics to determine changes in mRNA in reproductive tissues associated with preterm labour and preterm prelabour rupture of membranes; 3) proteomics to identify differentially expressed proteins in amniotic fluid of women with preterm labour; and 4) metabolomics to identify the metabolic footprints of women with preterm labour likely to deliver preterm and those who will deliver at term. The complementary nature of discovery science and HDB is emphasised.

Chronic hyperglycemia, early complications of pregnancy, and developmental abnormalities may be linkened by a disruption of normal development and function of the yolk sac. In primates, the yolk sac is immersed in the extra-embryonic celomic fluid (EECF). Our goal was to conduct a controlled study using an established timed-pregnant baboon model to determine if transitory exposures of the primate yolk sac and embryo to supra-physiologic concentrations of glucose were lethal. Ultrasound-guided celocentesis were performed in 7 baboons (3-control and 4-experimental) at 38–42 days from conception (term ∼182 days). EECF was partially replaced with a non-hyperglycemic solution in control and with a solution containing different glucose concentrations in study group (table). Final glucose concentration in EECF was calculated by the formula (EECF volume after aspiration × baseline celomic glucose concentration + volume of injected solution × glucose concentration)/final EECF volume. In baboons, baseline EECF glucose concentration at this developmental stage is 0.6 mg/mL. Estimated EECF volume was calculated by subtracting the volume of the amniotic sac and yolk sac from that determined for the gestational sac (each extra-embryonic structure was measured ultrasonographically and volume calculated by the formula = 4/3πr3). All animals delivered at term healthy newborns without developmental abnormalities. The table summarizes technical data at celocentesis. Final glucose concentration and pregnancy outcome are also shown in this table. This study has important clinical implications since it suggests that: 1) the primate yolk sac and embryo can tolerate transitory exposures to very high glucose concentrations without pregnancy complications; 2) chronic exposure to hyperglycemic states may be required to induce lethality or developmental complications.

The concentrations of pregnancy protein 1 (SP1), placental-specific tissue protein 10 (PP10), placental-specific tissue protein 12 (PP12) and alpha1-Fetoprotein (AFP) were analyzed in serum samples of 83 patients with bronchial carcinoma at stages II-IV Protein levels were determined by means of single radial immunodiffusion, rocket immuno-electrophoresis, radioimmunoassay and enzyme- immunoassay. PP12 and AFP serum concentrations were significantly increased in the cancer group compared with the control group. PP12 (control group: x=54.08 microg/l; s=61. 70; tumor group: x = 122.52 microg/l; s = 131.16); AFP (control group: x=3.05 microg/l; s=3. 76; tumor group: x = 8.29 microg/l; s = 17.75). SPI was found in only 22 cases of the tumor group. PP10 (control group: x = 2.25 microg/l; s = 0.866; tumor group: x = 2.503 microg/l; s=1.508). Given that at least two of the tested parameters were determined to be in the pathological range, the sensitivity amounted to 0.64 and the specificity to 0.92.