Estrogen- and Progesterone-Induced Variation in Corneal Parameters According to Hormonal Status.

OBJECTIVES: To determine the effects of estrogen and progesterone on corneal thickness, curvature, and biomechanics in healthy corneas according to hormonal status. METHODS: The study included four groups of females: group A (menstruating, not pregnant, not lactating, and not menopausal; n=100), group B (pregnant; n=50), group C (lactating; n=50), and group D (menopausal; n=50). Group A was subdivided according to age, as subgroup A15-25 (age 15-25 years) and subgroup A>25 (age >25 years). Blood estradiol and progesterone levels were measured in each participant. All the participants underwent a full ophthalmologic examination, including corneal thickness and corneal topography measurement, and evaluation of corneal biomechanical properties. RESULTS: The corneal resistance factor and anterior corneal flat keratometry values were significantly higher in group D (P=0.040 and P=0.026, respectively) than in the other three groups. Posterior corneal steep keratometry values were significantly higher in subgroup A>25 during the preovulatory phase than ovulatory and postovulatory phases (P=0.012). In group B, there was a significant negative correlation between gestational week and intraocular pressure (IOP) (r=-0.322, P=0.024). Corneal volume was significantly higher during the early postpartum period than the late postpartum period in group C (P=0.028). Intraocular pressure, Goldman-correlated IOP, and corneal-compensated IOP differed significantly between the groups (P<0.05). CONCLUSIONS: Blood levels of estrogen and progesterone were associated with variations in IOP, but estrogen and progesterone did not have a consistent effect on topographic parameters or biomechanical properties in healthy corneas.

The lipase cofactor CGI58 controls placental lipolysis.

In eutherians, the placenta plays a critical role in the uptake, storage, and metabolism of lipids. These processes govern the availability of fatty acids to the developing fetus, where inadequate supply has been associated with substandard fetal growth. Whereas lipid droplets are essential for the storage of neutral lipids in the placenta and many other tissues, the processes that regulate placental lipid droplet lipolysis remain largely unknown. To assess the role of triglyceride lipases and their cofactors in determining placental lipid droplet and lipid accumulation, we assessed the role of patatin like phospholipase domain containing 2 (PNPLA2) and comparative gene identification-58 (CGI58) in lipid droplet dynamics in the human and mouse placenta. While both proteins are expressed in the placenta, the absence of CGI58, not PNPLA2, markedly increased placental lipid and lipid droplet accumulation. These changes were reversed upon restoration of CGI58 levels selectively in the CGI58-deficient mouse placenta. Using co-immunoprecipitation, we found that, in addition to PNPLA2, PNPLA9 interacts with CGI58. PNPLA9 was dispensable for lipolysis in the mouse placenta yet contributed to lipolysis in human placental trophoblasts. Our findings establish a crucial role for CGI58 in placental lipid droplet dynamics and, by extension, in nutrient supply to the developing fetus.

Administration of steroids after 34 weeks of gestation enhances fetal lung maturity profiles.

OBJECTIVE: To estimate the effect of antenatal glucocorticoid administration on fetal lung maturity in pregnancies with known fetal lung immaturity between the 34th and 37th weeks of gestation. STUDY DESIGN: Pregnancies between 34(0/7) and 36(6/7) weeks undergoing amniocentesis to determine fetal lung maturity were targeted. Women with negative results (TDx-FLM-II <45 mg/g) were randomly assigned to intramuscular glucocorticoid injection or no treatment. A repeat TDx-FLM-II test was obtained 1 week after enrollment. RESULTS: Thirty-two women who met inclusion criteria were randomly assigned. Seven women delivered within a week of testing for fetal lung maturity, and were excluded from the analysis. Ten received glucocorticoid and 15 did not. Women assigned to glucocorticoids had a mean increase TDx-FLM-II in 1 week of 28.37 mg/g. Women assigned to no-treatment had an increase of 9.76 mg/g (P < .002). CONCLUSION: A single course of intramuscular glucocorticoids after 34 weeks in pregnancies with documented fetal lung immaturity significantly increases TDx- FLM-II.

Intrauterine Cataract Diagnosis and Follow-up

In this article, we report a 21-gestational-week fetus diagnosed with congenital cataract by ultrasonography. The parents decided to terminate the pregnancy and asked for examination of the fetus. An amniocentesis was performed for fetal karyotyping. After termination of the pregnancy, fetal autopsy was conducted. Whole exome sequencing (Trio-WES) analysis of the mother and father was done from peripheral blood samples. In the pathologic autopsy report, bilateral anterior and posterior subcapsular cataracts were confirmed. Whole exome sequencing analysis revealed a previously unreported class 3 variant of uncertain significance (c755A>G [P.Lys252Arg]) of the CRYBB1 gene, which is associated with congenital cataract, that was homozygous in the fetus and heterozygous in the parents. The obtained result is consistent with a genetic diagnosis of isolated autosomal recessive cataract.

Recanalization of Totally Occluded Stent of Superficial Femoral Artery with Multiple Direct Stent Puncture.

In-stent restenosis (ISR) is a frequent complication of endovascular stents implantation, especially in the superficial femoral artery (SFA). Beyond the standard interventions, direct stent puncture (DSP) to the totally occluded SFA increases the success rate of the endovascular procedures. Multiple attempts are required to treat total occlusions most of time. DSP useful and safe technique and provide good angiographic results. Beside the classical DSP, in this case we discussed recanalization of totally occluded stent of superficial femoral artery with bidirectional stent puncture.

PLIN2 Is Essential for Trophoblastic Lipid Droplet Accumulation and Cell Survival During Hypoxia.

Trophoblast hypoxia and injury, key components of placental dysfunction, are associated with fetal growth restriction and other complications of pregnancy. Accumulation of lipid droplets has been found in hypoxic nonplacental cells. Unique to pregnancy, lipid accumulation in the placenta might perturb lipid transport to the fetus. We tested the hypothesis that hypoxia leads to accumulation of lipid droplets in human trophoblasts and that trophoblastic PLIN proteins play a key role in this process. We found that hypoxia promotes the accumulation of lipid droplets in primary human trophoblasts. A similar accretion of lipid droplets was found in placental villi in vivo from pregnancies complicated by fetal growth restriction. In both situations, these changes were associated with an increased level of cellular triglycerides. Exposure of trophoblasts to hypoxia led to reduced fatty acid efflux and oxidation with no change in fatty acid uptake or synthesis. We further found that hypoxia markedly stimulated PLIN2 mRNA synthesis and protein expression, which colocalized to lipid droplets. Knockdown of PLIN2, but not PLIN3, enhanced trophoblast apoptotic death, and overexpression of PLIN2 promoted cell viability. Collectively, our data indicate that hypoxia enhances trophoblastic lipid retention in the form of lipid droplets and that PLIN2 plays a key role in this process and in trophoblast defense against apoptotic death. These findings also imply that this protective mechanism may lead to diminished trafficking of lipids to the developing fetus.

Hypoxia regulates the expression of fatty acid-binding proteins in primary term human trophoblasts.

OBJECTIVE: Fatty acids (FAs) are essential for fetal development. Cellular FA uptake is modulated by fatty acid-binding proteins (FABPs). We hypothesized that hypoxia regulates the expression of FABPs in human trophoblasts. STUDY DESIGN: Primary term human trophoblasts were cultured for 72 hours in either standard (O2 = 20%) or hypoxic (O2 < 1%) conditions. FABP expression was interrogated using polymerase chain reaction and Western immunoblotting. Trophoblast lipid droplets were examined using dipyrromethene boron difluoride 493/503 staining. RESULTS: We detected the expression of FABP1, -3, -4, -5, and pm but not FABP2 or FABP6-9 subtypes in trophoblasts. Exposure to hypoxia markedly increased lipid droplet accumulation in trophoblasts. Consistent with this observation, hypoxia enhanced the expression of FABP1, -3, and -4. Lastly, agonists of peroxisome proliferator-activated receptor-gamma enhanced the expression of FABP1 and -4 in trophoblasts. CONCLUSION: Hypoxia enhances the expression of FABP1, -3, and -4 in term human trophoblasts, suggesting that FABPs support fat accumulation in the hypoxic placenta.

Peroxisome proliferator-activated receptor-gamma and retinoid X receptor signaling regulate fatty acid uptake by primary human placental trophoblasts.

CONTEXT: Transplacental transfer of fatty acids from the maternal to the fetal circulation is essential for fetal development. The nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) regulates fatty acid transport and storage in adipocytes and other cell types. OBJECTIVE: This study tested the hypothesis that PPARgamma and its heterodimeric nuclear receptor partner, retinoid X receptor (RXR), regulate fatty acid uptake by human trophoblasts. DESIGN: Prospective basic laboratory in vitro research was conducted using primary term human trophoblasts. SETTING: The study was performed in the perinatal biology laboratory of an academic medical center. PATIENTS OR OTHER PARTICIPANTS: Study materials were obtained from healthy pregnant women at a gestational age of 37-41 wk. INTERVENTIONS: There were no interventions. MAIN OUTCOME MEASURES: Fat uptake and accumulation in human placental trophoblasts were measured. RESULTS: We initially demonstrated that activation of PPARgamma and/or RXR with selective agonists increased the accumulation of neutral lipids in trophoblasts as well as uptake of free fatty acids. Furthermore, activation of PPARgamma and RXR enhanced the expression of the fat droplet-associated protein adipophilin along with fatty acid transport protein (FATP)4, whereas expression of FATP2 was decreased by activation of RXR. Finally, we found that inhibition of p38 MAPK, which diminishes the activity of PPARgamma in trophoblasts, inhibited fatty acid uptake and blocked the PPARgamma- and RXR-dependent increases in adipophilin and FATP4 expression, yet stimulated the expression of FATP1, FATP2, and FATP3. CONCLUSIONS: These data support a role for PPARgamma and RXR in regulation of fatty acid transport and storage in human placental trophoblasts.

The mean weekly increment of amniotic fluid TDx-FLM II ratio is constant during the latter part of pregnancy.

OBJECTIVE: The purpose of this study was to determine the mean weekly increment in amniotic fluid TDx-FLM II ratio during the latter part of pregnancy. STUDY DESIGN: All women who underwent > 1 amniocentesis for the determination of fetal lung maturity between 1998 and 2004 were identified retrospectively. Clinical information and TDx-FLM II ratios were collected from the participant's chart and analyzed. RESULTS: The gestational age of all participants at the first TDx-FLM II test was 31.2 to 37.5 weeks (mean, 34.7 +/- 1.4 weeks of gestation). The median interval between the 2 tests was 7 days (range, 5-36 days). We found that the mean weekly increment of TDx-FLM II was 14.4 +/- 9.9 mg/g (surfactant to albumin) and remained constant across the gestational ages. CONCLUSION: The mean weekly increment of TDx-FLM II is 14.4 +/- 9.9 mg/g and is constant during the latter part of pregnancy. This information, combined with the gestational age, should be useful in treating women with an initial immature test.

Insulin and fatty acids regulate the expression of the fat droplet-associated protein adipophilin in primary human trophoblasts.

OBJECTIVE: This study was undertaken to test the hypothesis that insulin and fatty acids regulate adipophilin expression in cultured human trophoblasts. STUDY DESIGN: Cytotrophoblasts isolated from term human placentas were cultured in the absence or presence of insulin (10 nmol/L), and a mix of oleic and linoleic acid in serum-free medium. The expression of adipophilin as well as the fatty acid transport proteins (FATP) 2, 3, 4 and 6 was examined. Fat accumulation was quantified by BODIPY staining and fat uptake determined using [3H]-oleic acid. RESULTS: A combination of insulin and fatty acids enhanced the expression of adipophilin (2.3-fold, P < .05). In contrast, the expression of FATPs was unchanged. Furthermore, insulin and fatty acids increased the accumulation of fat droplets in trophoblasts by 4- to 5-fold (P < .05), but had no effect on oleic acid uptake. CONCLUSION: Insulin and fatty acids enhance the expression of adipophilin and the formation of fatty acid droplets in term human trophoblasts.

The lipid droplet-associated protein adipophilin is expressed in human trophoblasts and is regulated by peroxisomal proliferator-activated receptor-gamma/retinoid X receptor.

Uptake and transplacental transfer of fatty acids is essential for fetal development. Human adipophilin and its murine ortholog adipocyte differentiation-related protein are lipid droplet-associated proteins that are implicated in cellular fatty acid uptake in adipocytes. The nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARgamma) stimulates lipid uptake by adipocytes and enhances differentiation of placental trophoblasts. We therefore hypothesized that adipophilin is expressed in human trophoblasts and that its expression is regulated by PPARgamma. We initially determined that adipophilin is expressed in human villous trophoblasts and that adipophilin expression is enhanced during differentiation of cultured primary term human trophoblasts. We also found that exposure of cultured human trophoblasts to the PPARgamma ligand troglitazone resulted in a concentration-dependent increase in adipophilin expression. We observed a similar increase with LG268, a ligand for retinoid X receptor (RXR), the heterodimeric partner of PPARgamma. Lastly, we demonstrated that ligand-activated PPARgamma and RXR stimulated the transcriptional activity of adipophilin promoter in CV-1 cells and in the placental JEG3 cell line. We conclude that the expression of adipophilin is enhanced during trophoblast differentiation and is up-regulated by ligand-activated PPARgamma/RXR. Enhanced adipophilin expression may contribute to fatty acid uptake by the placenta.

Effects of melatonin on noncardiogenic pulmonary edema secondary to adnexial ischemia-reperfusion in guinea pig.

OBJECTIVES: Pulmonary edema has been shown to occur following ischemia-reperfusion injury in a variety of organs and effects of several pharmacological agents on ischemia-reperfusion-induced damage has been investigated previously. However, there are only a few studies in the literature about pulmonary injury following adnexial ischemia-reperfusion. In this study we aimed at investigating pulmonary changes following adnexial ischemia-reperfusion and the effects of melatonin on noncardiogenic pulmonary edema secondary to adnexial ischemia-reperfusion. METHODS: A total of 32 Dunkin-Harley guinea pigs were randomly divided into four groups. In group I, oopherectomy was performed following adnexial torsion of three hours (ischemia). Adnexial torsion of three hours followed by a 3 hours detorsion period (reperfusion) and then oopherectomy was performed in remaining three groups. No therapy was given in group one and two while isotonic saline and melatonin was applied intraperitoneally in groups three and four, respectively. Serum malondialdehyde (MDA) levels of operated ovaries and lungs and polimorphonuclear leucocyte infiltration of lungs were determined. RESULTS: MDA levels in serum, ovary and lungs were higher in detorsion groups (groups II, III) than torsion group (group I) (p<0.01). Melatonin administration significantly decreased the polimorphonuclear leukocytes infiltration of lung parenchime and MDA levels in serum, ovaries and lungs (groups II and IV; groups III and IV; p<0.01, p<0.01). MDA levels and lung tissue PNL infiltration levels of melatonin administered detorsioned group was similar to those levels of only salphingo-oopherectomy performed group (Group I and IV, p>0.05). CONCLUSION: Pathophysiology of ischemia-reperfusion must be considered in the cases of adnexial torsion where detorsion is thought. As an antioxidant, melatonin administration might be helpful in decreasing post-operative morbidity by decreasing reperfusion injury of lungs.

Fetal heart rate accelerations and the risk of cerebral lesions and poor neurodevelopmental outcome in very low birthweight neonates.

The risk of intraventricular hemorrhage and periventricular leukomalacia correlates with fetal brain immaturity. Given that the appearance of fetal heart rate (FHR) accelerations is associated with brain maturation, we tested the hypothesis that neonatal cerebral lesions and developmental delay in very low birthweight newborns are associated with absent reactivity of the FHR tracing prior to delivery. We analyzed the FHR tracing of 97 fetuses with birthweight < 1200 g who underwent head ultrasound at day 3 and Bayley Scales of Infant Development testing at age 1 year. We used multivariate analysis to adjust for confounding variables. We found that the absence of two FHR accelerations of 10 beats per minute (bpm) for 10 seconds twice in a 20-minute window 1 hour before delivery was associated with intraventricular hemorrhage and/or periventricular leukomalacia ( P < 0.01) and a significant risk for mental and psychomotor delays by Bayley testing ( P < 0.001). The absence of accelerations of 15 bpm for 15 seconds was not associated with these abnormalities. The absence of FHR accelerations before delivery suggests a greater risk for cerebral injury and developmental delay in the very premature neonate.

Removal of hydrosalpinges increases endometrial leukaemia inhibitory factor (LIF) expression at the time of the implantation window.

BACKGROUND: The presence of hydrosalpinges is associated with lower implantation and pregnancy rates in women undergoing IVF-embryo transfer, while salpingectomy improves these parameters. Although the mechanism by which hydrosalpinges affects fertility is not entirely understood, an adverse effect on endometrial receptivity has been postulated. In this study, we hypothesized that the adverse effects of hydrosalpinges on fertility may be in part mediated by inappropriate endometrial expression of leukaemia inhibitory factor (LIF), a cytokine implicated in implantation. METHODS: In order to test our hypothesis, we prospectively examined the expression of LIF during the window of implantation in the endometrium of infertile women (n = 10) with hydrosalpinges prior to and following salpingectomy and of fertile controls (n = 10) by Western blotting and immunohistochemistry. RESULTS: LIF expression was significantly lower in infertile women with hydrosalpinges compared with fertile controls (P < 0.05). Salpingectomy resulted in an increase in LIF expression in eight out of 10 women with hydrosalpinges. LIF levels were increased by 231 +/-49% (mean +/- SEM) following salpingectomy. Immunohistochemical analysis confirmed the Western blot findings. The increased LIF immunoreactivity was predominantly localized to luminal and glandular epithelial cells. CONCLUSIONS: Our findings suggest that observed benefit from salpingectomy in infertile women with hydrosalpinges may be in part mediated by the up-regulation of endometrial LIF expression.