Alessandra Corradetti

The involvement of inflammatory cytokines in the pathogenesis of recurrent miscarriage.

OBJECTIVE To investigate the inflammatory cytokine expression pattern in trophoblastic tissue from women with unexplained recurrent miscarriage (RM). STUDY DESIGN Trophoblasts were obtained during uterine evacuation from 11 women with RM and from 20 healthy pregnant women undergoing elective termination of pregnancy, who served as controls. The array was performed using GEArray Q Series Human Inflammatory Cytokines & Receptors Gene Array HS-015 membranes. Data were confirmed by quantitative real-time PCR. The Mann-Whitney U test was performed for statistical analysis. RESULTS Microarray analysis identified three genes that were differentially expressed between RM patients and controls. We observed significant downregulation of Transforming Growth Factor beta 3 (TGF-β3) and Interleukin 25 (IL-25) (5-fold reduction and 2.5-fold reduction, respectively) and significant upregulation of CD-25, also known as Interleukin 2 receptor alpha (IL-2RA) (7-fold increase) in women with RM compared with controls. The median ΔC(t) of TGF-β3 was 8.2 (interquartile range, 7.67-8.9) in RM patients vs. 5.85 (interquartile range, 5.3-6.09) in controls; the median ΔC(t) of IL-25 was 5.18 (interquartile range, 4.46-5.76) in RM patients vs. 3.85 (interquartile range, 3.6-4.51) in controls, and the median ΔC(t) of CD-25 was 9.62 (interquartile range, 7.81-12.42) in RM patients vs. 12.44 (interquartile range, 11.02-13.86) in controls. DISCUSSION Our results suggest that the immunological and inflammatory regulation mechanisms of the placental environment play a key role in recurrent miscarriage. The observed trophoblast cytokine expression pattern at the maternal-fetal interface confirms the immunotrophic theory, as demonstrated by a switch from a T-helper-1 (Th1) profile to a T-helper-2 (Th2) profile in women who experience recurrent miscarriages.

Perinatal outcomes in oocyte donor pregnancies

Abstract Objective: To assess the obstetric outcomes of pregnancy following intracytoplasmic sperm injection (ICSI) using donor oocytes. Methods: Twenty-six deliveries from oocyte donor ICSI (d-ICSI) were compared to the next two consecutive deliveries from homologous ICSI (h-ICSI group) (n = 52) and with the two consecutive deliveries from women older than 40 years (Advanced Maternal Age: AMA) (n = 52). We evaluated the occurrence of gestational hypertension (GH), preeclampsia (PE), fetal growth restriction (IUGR), gestational diabetes mellitus (GDM), preterm premature rupture of membranes (pPROM), preterm birth, placental anomalies, mode of delivery, hemorrhage, gestational age at birth and birth weight. Results: d-ICSI had significantly more PE (d-ICSI 19.2%, h-ICSI 0%, AMA 0%, p < 0.001); higher rates of IUGR than AMA pregnancies (d-ICSI 19.2%, AMA 3.8%, p < 0.025). Placental accretism was found only in the d-ICSI group (15.4%, p < 0.043). No postpartum bleeding was observed. Conclusions: This is the first study that compares the obstetric outcomes of donor pregnancies to the outcomes of h-ICSI and AMA. Obstetricians who deal with pregnancies from oocyte donation need to be aware of the more severe obstetric outcomes, especially placenta accreta and preeclampsia. All women who conceive through oocyte donation should be counseled as early as the pre-conception period and referred to specific centers for high-risk pregnancies.

Plasma lipids and physicochemical properties of the erythrocyte plasma membrane throughout pregnancy

Background.  The physiopathological relevance of plasma lipid concentrations is supported by the observation that they might affect the physicochemical properties of the plasma membrane of circulating cells and might be crucial in the pathological conditions complicating pregnancy.

The role of p38α mitogen-activated protein kinase gene in the HELLP syndrome

Mitogen-activated protein kinase (MAPK) p38α was shown to be implicated in the organogenesis of the placenta, and such placental alteration is crucial for the development of hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome. We aimed to analyze for the first time human placental expression of MAPK p38α in pregnancies complicated by HELLP. The placental expression of MAPK p38α was investigated by semiquantitative polymerase chain reaction using cDNA extracted from placental tissue of 15 pregnancies with HELLP syndrome and 15 gestational age-matched controls. Seven patients with HELLP also had intrauterine fetal growth restriction (IUGR). In placenta from pregnancy complicated by HELLP, the expression of MAPK p38α is significantly decreased compared to the group with normal pregnancy (p < 0.001), while no difference was found between the HELLP and HELLP with IUGR subpopulations. Our study shows for the first time that MAPK p38α is expressed in the human placenta. Pregnancies with placental dysfunction and hypertensive complications are characterized by a significantly decreased expression of MAPK p38α. Our observations suggest that p38 MAPK signaling may be essential in placental angiogenesis and functioning.

Alpha-hemoglobin-stabilizing protein (AHSP) in hemolysis, elevated liver enzyme, and low platelet (HELLP) syndrome, intrauterine growth restriction (IUGR) and fetal death.

ObjectiveAlpha hemoglobin-stabilizing protein (AHSP) inhibits the production of reactive oxygen species in various cells, including erythrocytes. Reduced AHSP can mean reduced protection from stressors. Our objective was to investigate whether AHSP is involved in the response to stress in pregnancy.Study designPlacentas were collected from normal term pregnancies (n = 10) and pregnancies complicated by HELLP (n = 10), intrauterine growth restriction (IUGR; n = 10) or fetal death (IUFD; n = 6). AHSP messenger RNA (mRNA) and protein were determined using real time quantitative polymerase chain reaction (PCR) and Western blot, respectively. All statistical analyses were performed by using the GraphPad Prism Software. Differences were considered significant at p < 0.05.ResultsPlacental AHSP mRNA level in HELLP (4.16E10−4 ± 1.77) and IUFD (4.19E10−4 ± 3.37) were significantly decreased compared with controls (28.47E10−4 ± 14.86; p < 0.01), whereas levels in the IUGR group (7.55E10−4 ± 6.4) showed a trend toward being lower but the difference did not reach statistical significance. Western blot analysis results indicate a no significant increase of ASHP protein in the HELLP syndrome group and a significant decrease in the IUFD group compared with controls. There was no significant difference between the IUGR and control groups.ConclusionASHP mRNA expression in the placenta is decreased in complicated pregnancies, and it may be involved in the pathogenic mechanisms leading to the adverse pregnancy outcome.

Congenital Uterine Malformations Are Associated to Increased Blood Pressure in Pregnancy

Objective: To assess the relationship between congenital uterine malformations and blood pressure in pregnancy. Study Design: Twenty‐four‐hour automated ambulatory blood pressure monitoring (readings every 30 min) was performed in 16 normotensive, nonproteinuric, primigravidae with congenital uterine malformations (5 uterus septus, 9 uterus bicornis, 2 uterine didelphys) between 20 and 30 weeks. From the 24‐hr blood pressure report, we calculated 24‐hr mean, daytime and nighttime means. The results were compared with 16 primigravidae, matched for age and gestation, who were and remained normotensive throughout pregnancy, and tested for statistics with t‐test; significance assessed at p < 0.001. Results: Although they were within the normotensive range, all blood pressure measurements considered were significantly higher in pregnant women with congenital uterine malformations, compared to normal pregnant women. Namely, 24‐hr, daytime, and nighttime systolic (mean ± SD) were 121.1 ± 8.4, 124.4 ± 8.8, 114.0 ± 7.7 mmHg, respectively, in women with uterine malformations and 108.0 ± 7.4, 109.2 ± 7.3, 102.1 ± 8.5 mmHg, respectively, in normal pregnant women. Twenty‐four‐hour diastolic, daytime, and nighttime diastolic (mean ± SD) 74.1 ± 10.2, 77.1 ± 10.6, 68.1 ± 9.2 mmHg, in women with uterine malformations and 64.1 ± 5.7, 66.0 ± 5.7, 58.2 ± 6.3 mmHg, in normal pregnant women (all differences p < 0.001). Fifteen of the fetuses from women with congenital uterine malformations showed intrauterine growth retardation. No differences were found 6 months after delivery. Conclusions: Although the blood pressure levels remained within the normotensive range, pregnant women with congenital uterine anomalies have a higher blood pressure than normal women. Elevated blood pressure can result from altered uterine circulation and reduced blood supply to the placenta. This pathogenesis or the poor placentation may result in a foetal growth retardation.

PP035. Placental klotho protein in preeclampsia: A possible link to long term outcomes

INTRODUCTION An aging-suppressor gene, klotho, is a candidate factor for vascular disease because its deficiency leads to impaired endothelium-dependent vasodilation and impaired angiogenesis. Although klotho protein is predominantly expressed in the kidney, it is detected in a limited number of other tissues, such as the placenta, ovary, prostate gland, and small intestine. This protein is involved in several metabolic pathways such as calcium and phosphate homeostasis, the insulin-like growth factor 1 (IGF-1), apoptosis, angiotensin-II-induced events in the kidney and oxidative stress. OBJECTIVES The aim was to assess the expression of the klotho gene in the placenta from pregnancies affected by severe preeclampsia. METHODS Placentas were collected from normal pregnancies (n=12) and pregnancies complicated by preeclampsia (n=12), matched for gestational age. Klotho mRNA and protein were determined using real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. RESULTS Real-Time PCR analyses demonstrated a significant (p=0.005) 83% down-regulation of Klotho in patients with Preeclampsia versus Controls. Results of Western Blot agreed with those from Real-Time PCR. CONCLUSION Klotho mRNA expression in the placenta is decreased in preeclamptic pregnancies. Given its role in cardiovascular disease in aging, it may link preeclamptic mothers and their offsprings to long term cardiovascular outcomes.

Placental Cytokines in the Pathogenesis of Preeclampsia and Hellp Syndrome

Preeclampsia and HELLP syndrome are placenta-dependent disorders with both local and systemic anomalies that are responsible for neonatal and maternal morbidity. The cytokines, produced by the placenta in response to local ischemia/hypoxia, may be involved in endothelial activation and dysfunction. The adaptive immune system could play a key role in the etiology of preeclampsia or HELLP by generating a pro- inflammatory Th1 type immune reaction: the current pathophysiologic hypothesis of preeclampsia is focused on maladaptation of immune responses and defective trophoblast invasion. Also human decidual NK cells recruited at the site of embryonic implantation induce a number of cytokines with potential functions not yet clearly established. We reviewed recent studies on effect of pro-inflammatory cytokines in preeclampsia, as well as on the role of regulatory cytokines and chemokines and discuss evidence that cytokines continue to be part of a paracrine/autocrine regulatory network in the placenta and membranes throughout the different stages of gestation. In addition we reviewed the experimental basis for the possible role of the immune system and proposed the hypothesis that these conditions could be a placental inflammatory response which can lead to a systemic and endothelial dysfunction, resulting in hypertension, proteinuria and pathologies in many organ systems.

PP093 Maternal and fetal outcomes in oocyte donor pregnancies

INTRODUCTION The history of oocyte donation is relatively new in the framework of in vitro fertilization (IVF) techniques, and little has been discussed about the obstetric outcomes of such pregnancies. OBJECTIVES The aim of this study is to assess the obstetric outcomes of pregnancy following in vitro fertilization with embryo transfer (IVF-ET) using donor oocytes and compare them to the outcomes from autologous IVF-ET and to spontaneous pregnancy in women with advanced age (AMA) to identify possible criticalities and help in counseling women and their doctors. METHODS The study included a total of 70 delivered pregnancies. The study group included 14 oocyte donors IVF-ET (d-IVF-ET) from women aged 32-52years. The results from the study group were compared to the next two consecutive deliveries from the autologous IVF-ET (IVF-ET group) (n=28; age 30-46years) and with two more consecutive deliveries from women older than 40years (Advanced Maternal Age: AMA) (n=28, age 40-45years). We evaluated the occurrence of pregnancy-induced hypertension (PIH), preeclampsia (PE), fetal growth restriction (IUGR), the gestational age at birth, placental anomalies, the mode of delivery, birth weight and the neonatal Apgar score. The fetal weight was corrected with the gestational age at the time of delivery according to Gardosi. Statistical analysis was performed with the Chi-squared test. RESULTS Oocyte donor pregnancies had significantly higher rates of PE (d-IVF-ET 21.4%, IVF-ET 0%, AMA 0%, p<0.011). They also had higher rates of PIH and IUGR (d- IVF-ET 21.4%, IVF-ET 0%, AMA 3.6% p<0.011) (d- IVF-ET 21.4%, IVF-ET 7.1%, AMA 3.6% p<0.011 respectively). We found placental anomalies only in the d-IVF-ET group; the incidence of placental accretism was 28.6%, (p<0.003). There are not significant differences in the gestational age at birth, placental anomalies, the mode of delivery, birth weight and the neonatal Apgar score between the groups. CONCLUSION This is the first study that compares the obstetric outcomes of donor pregnancies to the outcomes of autologous IVF-ET pregnancies and to advanced maternal age. The advanced maternal age criterion assumes that most women requiring oocyte donation are older. Hypertensive disorders were surprisingly not related to maternal age or to the in vitro fertilization technique. Obstetricians that deal with pregnancies from oocyte donation need to be aware of the more severe obstetric outcomes, especially placenta accrete and pregnancy-related hypertensive disorders. This warrants close blood pressure monitoring and an accurate placenta ultrasound. All women who conceive through oocyte donation should be counselled as early as the pre-conception period and referred to specific centres for high-risk pregnancies.