Mitochondrial Content and Hepcidin are Increased in Obese Pregnant Mothers

OBJECTIVE: Maternal obesity is characterized by systemic low-grade inflammation and oxidative stress (OxS) with the contribution of fetal sex dimorphism. We recently described increased mitochondrial content (mtDNA) in placentas of obese pregnancies. Here, we quantify mtDNA and hepcidin as indexes of OxS and systemic inflammation in the obese maternal circulation. METHODS: Forty-one pregnant women were enrolled at elective cesarean section: 16 were normal weight (NW) and 25 were obese (OB). Obese women were further classified according to the presence/absence of maternal gestational diabetes mellitus (GDM); [OB/GDM(-)]: n\u2009=\u200915, [OB/GDM(+)]: n\u2009=\u200910. mtDNA and hepcidin were evaluated in blood (real-time PCR) and plasma (ELISA). RESULTS: mtDNA and hepcidin levels were significantly increased in OB/GDM(-) versus NW, significantly correlating with pregestational BMI. Male/female (M/F) ratio was equal in study groups, and overall F-carrying pregnancies showed significantly higher mtDNA and hepcidin levels than M-carrying pregnancies both in obese and normal weight mothers. CONCLUSIONS: Our results indicate a potential compensatory mechanism to increased obesity-related OxS and inflammation, indicated by the higher hepcidin levels found in obese mothers. Increased placental mitochondrial biogenesis, due to lipotoxic environment, may account for the greater mtDNA amount released in maternal circulation. This increase is namely related to F-carrying pregnancies, suggesting a gender-specific placental response

Impact of Obesity and Hyperglycemia on Placental Mitochondria

A lipotoxic placental environment is recognized in maternal obesity, with increased inflammation and oxidative stress. These changes might alter mitochondrial function, with excessive production of reactive oxygen species, in a vicious cycle leading to placental dysfunction and impaired pregnancy outcomes. Here, we hypothesize that maternal pregestational body mass index (BMI) and glycemic levels can alter placental mitochondria. We measured mitochondrial DNA (mtDNA, real-time PCR) and morphology (electron microscopy) in placentas of forty-seven singleton pregnancies at elective cesarean section. Thirty-seven women were normoglycemic: twenty-one normal-weight women, NW, and sixteen obese women, OB/GDM(-). Ten obese women had gestational diabetes mellitus, OB/GDM(+). OB/GDM(-) presented higher mtDNA levels versus NW, suggesting increased mitochondrial biogenesis in the normoglycemic obese group. These mitochondria showed similar morphology to NW. On the contrary, in OB/GDM(+), mtDNA was not significantly increased versus NW. Nevertheless, mitochondria showed morphological abnormalities, indicating impaired functionality. The metabolic response of the placenta to impairment in obese pregnancies can possibly vary depending on several parameters, resulting in opposite strains acting when insulin resistance of GDM occurs in the obese environment, characterized by inflammation and oxidative stress. Therefore, mitochondrial alterations represent a feature of obese pregnancies with changes in placental energetics that possibly can affect pregnancy outcomes

Placental ERRγ-CYP19 expressions and circulating 17-Beta Estradiol in IUGR pregnancies

Introduction: Sex steroids are regulating factors for intrauterine growth. 17- f Estradiol (E2) is particularly critical to a physiological pregnancy, as increased maternal E2 was correlated to lower birth weight. The placenta itself is a primary source of estrogens, synthetized from cholesterol precursors. Cytochrome P450 aromatase (encoded by CYP19 gene) is a rate-limiting enzyme for E2 biosynthesis. CYP19 transcription is supported by Estrogen Related-Receptor Gamma (ERR\u3b3), thus having an indirect role in placental steroidogenesis. Here we investigated maternal E2 levels and placental CYP19 and ERR expressions in pregnancies with intrauterine growth restriction (IUGR). Methods: Singleton pregnancies were studied. E2 was measured in maternal plasma by electrochemiluminescence in 16 term controls and 11 IUGR (classified by umbilical artery doppler Pulsatility Index) at elective cesarean section, and also in 13 controls during pregnancy at a gestational age comparable to IUGR. CYP19 and ERR\u3b3 expressions were analyzed in placental tissue. Maternal/fetal characteristics, placental and molecular data were compared among study groups and tested for correlations. Results: Maternal E2 plasma concentrations were significantly decreased in IUGR compared to controls at delivery. When analyzing normal pregnancies at the same IUGR gestational age, E2 levels were not different to IUGR. However, E2 levels at delivery positively correlated with placental efficiency. Placental CYP19 was significantly higher in IUGR placental tissue versus controls, with levels specifically increased in female IUGR placentas. ERR\u3b3 expression was not different among groups. Discussion: We report a positive correlation between 17- f Estradiol levels and placental efficiency, that might indicate a disrupted steroidogenesis in IUGR pregnancies. Moreover, we showed alterations of CYP19 in IUGR placentas, possibly indicating a compensatory effect to the adverse IUGR intrauterine environment, also depending on fetal sex. Further studies are needed to deeper investigate IUGR alterations in the complex interaction among molecules involved in placental steroidogenesis

Sex specific adaptations in placental biometry of overweight and obese women

Introduction: Placental biometry at birth has been shown to predict chronic disease in later life. We hypothesized that maternal overweight/obesity, a state of low-grade inflammation and risk factor for adverse pregnancy outcome, could negatively influence placental development and that differences would be sex-specific. Methods: 696 women (537 normal-weight, NW; 112 overweight, OW; 47 obese, OB) with singleton uncomplicated pregnancies were prospectively enrolled at term delivery. Gestational age, maternal (age, height, pre-pregnancy BMI, gestational weight gain -GWG, hemoglobin, hematocrit and glycemia), fetal (weight, length, ponderal index, cranial circumference) and placental (weight, diameters) data were collected. Placental area, thickness and efficiency (fetal/placental weight ratio, F/P) were calculated. Results: GWG was within standard recommendations in OB, while OW exceeded it. Placental weight was significantly higher in OW versus NW, but not in OB, leading to significantly higher placental thickness and lower F/P in this group. In the total population, a significant interaction effect between maternal BMI and fetal sex on placental weight and efficiency was found. Indeed, differences in placental parameters were present only in female offspring. Discussion: In our population of OW and OB uncomplicated pregnancies only OW women, presenting GWG over standard recommendations, had thicker and less efficient placentas. We also reported different placental adaptation depending on fetal sex, with significant changes only in female fetuses. This may be part of a female-specific strategy aiming to ensure survival if another adverse event occurs. Customized counseling according to maternal BMI and fetal sex should be evaluated in clinical care

17-Beta Estradiol in obese pregnancies

INTRODUCTION Maternal obesity (MO) impacts on pregnancy and fetal outcomes, possibly altering intrauterine programming leading to adulthood diseases. Its energetic imbalance results in increased circulating fatty acids and consequent inflammation and oxidative stress. MO has been associated to both systemic and hormonal changes, but the metabolic impact of excessive fatty acids on pregnancy is not fully understood. Estrogens physiologically regulating pregnancy-related insulin resistance may also exacerbate obesity-related inflammation. During pregnancy the fetal-placental unit becomes a primary source of estrogens, particularly of 17-Beta Estradiol (E2). An obesity-related impairment of placental steroidogenesis has been reported. We measured maternal plasma E2 in relation to pregestational BMI and gestational diabetes mellitus (GDM). METHODS Venous blood was collected at elective cesarean section from 24 normal-weight (NW) and 23 obese (OB) women, 8 with GDM [OB/GDM(+)] (75 gr-OGTT; FIGO guidelines). EDTA samples were centrifuged at 1,500 rpm x 15 min and plasma selected excluding hemolyzed, icteric and lipemic. Samples were diluted 1:10 and run in duplicate on Cobas e411 to measure E2 concentration by an electrochemiluminescence immunoassay. Clinical and molecular data were analyzed with t-test and Pearson correlation. RESULTS When comparing to NW, maternal BMI was significantly different in OB (p 64 0.001), while basal glycaemia only in OB/GDM(+) (p 64 0.001) following inclusion criteria. Placental weight and thickness were significantly higher in both OB groups vs NW (p < 0.01), while efficiency (fetal/placental weight) was decreased in OB [6.68 \ub1 1.07] (p < 0.01) vs NW [8.01 \ub1 2.03]. E2 concentration [pg/mL] resulted significantly lower in OB [17,593.2 \ub1 5,493.6] vs NW [23,049.8 \ub1 11,810.1] (p 64 0.05). When considering the presence of GDM, OB/GDM(+) [19,701.9 \ub1 4,583.9] showed no differences compared to either OB/GDM(-) or NW, while OB/GDM(-) [16,468.5 \ub1 5,746.6] confirmed significantly lower E2 plasma concentration vs NW (p < 0.05). E2 levels correlated negatively with maternal BMI (p = 0.04, r = -0.30) and positively with placental efficiency (p = 0.01, r = +0.36) (Fig. 1). CONCLUSIONS Our preliminary analyses support evidences linking excessive BMI to decreased plasma E2, possibly impacting pregnancy outcomes. Indeed, E2 exerts a protective role against oxidative-stress, and obese lipotoxic environment can lead to decreased placental efficiency. GDM metabolic impairments related to insulin-resistance might represent an additional-opposing factor to the obese context, leading to increased E2 levels. Experiments on placental Estrogen Receptors (ER) will investigate a causal link to plasma E2 variation. Exploring the obesity-related effect on placental estrogen pathways could open future therapeutic features

Influence of Maternal Stress during Pregnancy in Females Born Small on F2 Male Metabolic and Cardiovascular Outcomes.

BACKGROUND We have previously shown that IntraUterine Growth Restriction (IUGR), is characterized by higher placental mitochondrial (mt) DNA levels than controls (C), and that this increase is negatively related with umbilical venous oxygenation. Here, we evaluate mt oxygen (O2) consumption and respiratory chain complexes (RCC) gene expression, in trophoblast cells isolated from IUGR and preeclamptic (PE) placentas. METHODS Cytotrophoblast cells were isolated from 16 placentas (8 IUGR, 3 PE alone, 5 C) of non-smoking women at elective caesarean section, and characterized by cytofluorimetry using cytokeratin-7 and anti-vimentin antibodies. mRNA levels of NDUFA9 (CI), ETFDH (CII), UQCRC1 (CIII) and COX4I1 (CIV) were quantified by Real Time (RT) PCR. O2 consumption was evaluated by High Resolution Respirometry (HRR), by administration of substrates and inhibitors of different RCC, thus allowing the measure of the global cell and of single complexes activity. Data were normalized by mtDNA content. RESULTS IUGR presented significantly lower UQCRC1 and COX4I1 mRNA levels than C (p<0.05). On the contrary, both raw (figure 1A) and normalized data (figure 1B) in IUGR with or without PE (but not in PE without IUGR) showed significantly higher O2 consumption levels of RCC, altogether and singularly, vs C (p<0.05 and p<0.01), particularly for CIV. In the entire population most of HRR data negatively related with ga and with placental and fetal weights. DISCUSSION Our data show lower mRNA levels of CIII and CIV subunits in trophoblast cells from insufficient placentas of IUGR fetuses, in contrast with the RCC higher functionality, suggesting a compensatory mechanism to their lower expression. These results shed new light into placental oxygenation in IUGR, suggesting that increased placental oxygen utilization may represent a limiting step in fetal growth restriction

The Role of Staphylococcus aureus in Mastitis: A Multidisciplinary Working Group Experience

Background: Breastfeeding women are at risk of developing mastitis during the lactation period. Staphylococcus aureus has emerged as the community-acquired pathogen responsible for virulence (methicillin resistance and Panton-Valentine leukocidin toxin producing). Research aim: The aim was to compare the microorganisms responsible for mastitis and breast abscesses during breastfeeding. Methods: This observational study was conducted with a sample of women (N = 60) admitted to our hospital between 2016 and 2018. Participants affected by mastitis and breast abscess were studied and cared for by a multidisciplinary working group. A diagnostic breast ultrasound identified the pathology. Results: Twenty-six participants (43.3%) were affected by mastitis and 34 (56.7%) by breast abscess. The most common microorganism identified was Staphylococcus aureus (S. aureus; mastitis, n = 13; abscesses, n = 24). Methicillin resistance was identified in 21 (44.7%) S. aureus strains: 17 (80.9%) cases of abscess and four (19.1%) cases of mastitis. The median number of months of breastfeeding was smaller in the methicillin-resistant S. aureus (MRSA) cases (median = 3, range = 1\u201320 months) than in the methicillin-sensitive S. aureus (MSSA) cases (median = 6.5, range = 3\u201321 months). The Panton-Valentine leukocidin toxin gene was detected in 12 (25.5%) cases (MRSA, n = 8, 66.7%; MSSA, n = 4, 33.3%). Hospitalization was required more frequently in MRSA (n = 8, 38%; five Panton-Valentine leukocidin positive) than in MSSA cases (n = 5, 19%; one Panton-Valentine leukocidin positive). Four women out of the eight MRSA cases (50%) that were Panton-Valentine leukocidin positive stopped breastfeeding during mammary pathologies, three (37.5%) participants continued breastfeeding until the follow-up recall, and one case was lost at follow-up. Conclusion: Clinical severity was probably complicated by the presence of the Panton-Valentine leukocidin toxin, which required hospitalization more frequently