Expression of extracellular matrix and adhesion proteins in pelvic organ prolapse.

Pelvic organ prolapse (POP) is a common disorder in women. It is characterized by the descent of the vaginal wall with consequent drop of pelvic organs. Pregnancy, labour and childbirth seem to be important events leading to the development of POP, since they are associated with prolonged stretch and mechanical stress of muscles, ligaments and connective tissue supporting pelvic organs. In pubocervical fascia, we explored the expression level of extracellular matrix and adhesion molecules. Tissue samples were obtained from twenty patients with POP who underwent cystocele repair, and from twenty control subjects during hysterectomy surgery. The PCR array analysis was performed and data were confirmed by Real-Time PCR and Western Blot.  Real-Time PCR results showed a significant upregulation for extracellular matrix protein 1 (ECM1) and integrin beta 3 (ITGB3) and a significant downregulation for FBLN5 in POP group. The decreased mRNA expression of FBLN5 in pathological samples was paralleled by a quantitative decrease in the corresponding protein, as Western Blot test highlighted. Our data provide an understanding of molecular mechanisms involved in POP-related pathophysiological processes and might represent an important tool to develop novel therapeutic agents for the treatment of this condition.

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-ß3) 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-ß3 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.

Placental Alpha Hemoglobin Stabilizing Protein (AHSP) and recurrent miscarriage.

AHSP inhibits cellular production of the reactive oxygen species. Reduced AHSP indicates reduced protection against oxidative stressors. Our objective was to investigate AHSP levels in recurrent miscarriage (RM). Trophoblast was collected from women of 10 weeks gestation: voluntary abortion controls (VA, n = 10); spontaneous first miscarriage with subsequent normal pregnancy (SMSN, n = 15) or with subsequent miscarriage (SMSM, n = 5); RM previously investigated (RMPS, n = 5) or not previously investigated (RM, n = 5). AHSP mRNA and protein were determined using real-time quantitative polymerase chain reaction (PCR) and Western blot, respectively. One-way ANOVA was performed to assess statistical significance (p < 0.05). ahsp mRNA levels were maximally reduced in RM and RMPS (8.0 x 10(-6) +/- 1.3 and 8.1 x 10(-6) +/- 0.7, respectively) compared with SMSN and VA (16.1 x 10(-6) +/- 2.3 and 26.1 x 10(-6) +/- 2.7, respectively). SMSM showed levels significantly reduced as well (9.0 x 10(-6) +/- 2.3). In RM, a reduced defense from oxidative stressors is evident at first miscarriage, identifying women at high risk for subsequent eventful pregnancy. Reduced AHSP may identify women at risk of experiencing further miscarriages.

Inflammatory cytokines patterns in the placenta of pregnancies complicated by HELLP (hemolysis, elevated liver enzyme, and low platelet) syndrome.

OBJECTIVE: We investigated the expression pattern and the role of inflammatory cytokines and their receptors in the placentas of pregnancy with HELLP syndrome. STUDY DESIGN: Placentas were collected after cesarean section, 10 from normal pregnancy and 10 from HELLP. The array was performed with GEArray Q Series Human Inflammatory Cytokines & Receptors Gene Array HS-015. The data were confirmed by quantitative real-time PCR. The Student's t test was used for statistical analysis. RESULTS: Macroarray analysis identified 14 cytokines differentially expressed. PCR confirmed that only IL-10, IL-6-receptor, and TGF-beta3 were increased, whilst CCL18, CXCL5, and IL-16 were significantly decreased, in HELLP. CONCLUSION: The regulation of cytokines involved in angiogenesis and adaptive immune responses may be critical for the placental vascular dysfunction. Our data support the hypothesis that HELLP syndrome could be a placental inflammatory response which leads to a systemic and endothelial dysfunction.

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.

Perinatal outcomes in oocyte donor pregnancies.

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.

The role of p38alpha mitogen-activated protein kinase gene in the HELLP syndrome.

Mitogen-activated protein kinase (MAPK) p38alpha 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 p38alpha in pregnancies complicated by HELLP. The placental expression of MAPK p38alpha 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 p38alpha 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 p38alpha is expressed in the human placenta. Pregnancies with placental dysfunction and hypertensive complications are characterized by a significantly decreased expression of MAPK p38alpha. 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.

OBJECTIVE: Alpha 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 DESIGN: Placentas 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. RESULTS: Placental 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. CONCLUSION: ASHP 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.

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. METHODS: Plasma and erythrocyte membrane lipid composition, membrane fluidity and function [membrane-bound enzyme sodium/potassium adenosine triphosphatase (Na+/K+-ATPase) activity] were studied in 24 healthy women in the nonpregnant condition and at 12, 24 and 38 weeks' gestation. RESULTS: The plasma showed an increase in total and high density lipoprotein (HDL) cholesterol, triglyceride (TG) and phospholipid (PL) levels. In the erythrocyte membrane we found the cholesterol/phospholipid (C/PL) ratio increased and the saturated to unsaturated fatty acid (FA) ratio (Sat/Unsat) significantly reduced; fluorescence polarization showed an initial increase during pregnancy and a progressive decrease afterwards; Na+/K+-ATPase activity was progressively reduced. CONCLUSIONS: A generalized alteration in the composition and function of the maternal erythrocyte membrane is present during a physiological pregnancy, and these modifications may involve a more complex physiopathogenetic mechanism.