Eran Barzilay

Uncoupling of brefeldin a-mediated coatomer protein complex-I dissociation from Golgi redistribution.

The Golgi complex functions in transport of molecules from the endoplasmic reticulum (ER) to the plasma membrane and other distal organelles as well as in retrograde transport to the ER. The fungal metabolite brefeldin A (BFA) promotes dissociation of ADP‐ribosylation‐factor‐1 (ARF1) and the coatomer protein complex‐I (COP‐I) from Golgi membranes, followed by Golgi tubulation and fusion with the ER. Here we demonstrate that the cationic ionophore monensin inhibited the BFA‐mediated Golgi redistribution to the ER without interfering with ARF1 and COP‐I dissociation. Preservation of a perinuclear Golgi despite COP‐I and ARF1 dissociation enables addressing the involvement of these proteins in anterograde ER to Golgi transport. The thermo‐reversible folding mutant of vesicular stomatitis virus G protein (VSVGtsO45) was retained in the ER in the presence of both monensin and BFA, thus supporting ARF1/COP‐I participation in ER‐exit processes. Live‐cell imaging revealed that BFA‐induced Golgi tubulation persisted longer in the presence of monensin, suggesting that monensin inhibits tubule fusion with the ER. Moreover, monensin also augmented Golgi‐derived tubules that contained the ER‐Golgi‐intermediate compartment marker, p58, in the absence of BFA, signifying the generality of this effect. Taken together, we propose that monensin inhibits membrane fusion processes in the presence or absence of BFA.

GnRH agonist vs. hCG for triggering of ovulation--differential effects on gene expression in human granulosa cells.

Objective To investigate the mRNA expression of genes related to steroidogenesis and OHSS in granulosa cells (GCs) of patients triggered with GnRH agonist compared to patients triggered with hCG. Design Mural GCs were obtained at the time of oocyte retrieval and gene expression was analyzed using quantitative real time RT-PCR. Settings Single center, case control study. Patient(s) 24 women who were treated with GnRH agonist or hCG for triggering of ovulation. Interventions GC collection. Main Outcome Measure(s) The expression of genes related to steroidogenesis and OHSS in mural GCs Results The fertilization rate was similar in the two groups. The mRNA expression of CYP19A1 (0.50 vs 1, arbitrary unit), CYP11A1 (0.6 vs. 1) and 3 beta hydroxysteroid-dehydrogenase (0.39 vs 1) was significantly lower in the GnRH group. The expression of VEGF (0.74 vs. 1) and inhibin β B (0.38 vs 1) was lower in the GnRH analog triggered group. Conclusion Expression of genes related to steroidogenesis is lower at the time of oocyte retrieval in patients triggered with GnRH agonist. The decreased expression of VEGF and inhibin β B in the GnRH agonist group can explain the mechanism of early OHSS prevention.

Erythropoietin Induced Tumour Mass Reduction in Murine Lymphoproliferative Models

leukocyte (PBL) counts begin to increase a few weeks following injection, exceeding 20 ! 10 6 cells/ml. Mice were injected (subcutaneously) with 10 4 MOPC-315 tumour cells in the abdomen area as described [7] . The rHuEPO (epoetin alfa, Eprex®; Janssen-Cilag, Baar, Switzerland) treatment (rHuEPO 30 U) was administered daily for 10 consecutive days, followed by three times per week for an additional 2–3 weeks to the MOPC-315-bearing mice as described [7] . The linear mixed effect (repeated measures) model was used to determine the effect of EPO on tumour size. Variance among mice was taken into account in the analysis by considering the mice to be randomly selected from a larger population. Starting 9 days after tumour cell injection, 28 MOPC315-bearing mice were injected with rHuEPO or albumin (control). The survival curve, shown in fi gure 1 d, demonstrates approximately 50 and 15% survival of the EPOtreated and control mice, respectively, which is in line with our previously documented observations [7] . Tumour growth kinetics in each of the EPO-treated and albumin-treated mice are presented in fi gures 1 a and b, respectively. Statistical analysis of the tumour size of 7 EPO-treated and 11 control progressor mice, which displayed only one localized tumour, using the linear mixed effect (repeated measures) model, yielded an R value of 0.875 ( fi g. 1 c). The predicted mean rate of tumour growth was 0.539 and 1.238 mm/day for EPO and albumin-treatRecombinant erythropoietin (rHuEPO) is widely used in clinical practice in the treatment of several types of anaemia [1–4] . We observed that patients with end-stage multiple myeloma (MM) treated with EPO live longer than expected, despite their original poor prognostic features [5, 6] . BALB/c mice in which MM was induced by transplantation of mineral oil-induced plasmacytoma cells (MOPC-315) showed complete T cell-mediated tumour regression in 30–60% of the animals after treatment with EPO [7] , suggesting that EPO may also act as an antitumour immunotherapeutic agent. Here, we raised a question regarding the effect of EPO on tumour load, rather than on the ultimate survival, by studying two lymphoproliferative murine models, MOPC-315 MM [8] and B cell leukaemia/lymphoma (BCL1) [9] . BCL1 is a B-cell leukaemia/lymphoma that developed spontaneously in a 2-year-old female BALB/c mouse [10] ; its advantage as a model is due to its analogies to human chronic lymphocytic leukaemia/lymphoma. In both models, we focused on tumour-bearing mice that did not achieve complete tumour regression after EPO treatment (‘progressors’). Female inbred BALB/c mice, aged 6–8 weeks, were obtained from the Tel-Aviv University Breeding Center. BCL1 tumour cells (10 4 cells), derived from spleens of tumour-bearing BALB/c mice, were injected intra-peritoneally into syngeneic mice. Typically, peripheral blood Received: January 18, 2005 Accepted after revision: May 9, 2005

Establishment and validation of a model for non-luteinized human mural granulosa cell culture.

Cell culture techniques of human mural granulosa cells (MGCs) serve as a major in vitro tool. However, the use of luteinized MGCs has major limitations due to their luteinized state. Our aim was to establish a standardized protocol for the culture of MGCs as a model for different stages of folliculogenesis. We showed that early-non-luteinized, preovulatory-non-luteinized and luteal-MGCs have distinct gene expression pattern. After 4 days of incubation of luteinized-MGCs, ovulatory genes mRNAs achieve expression levels similar to the early non-luteinized follicles. FSH stimulation for 48 h of these 4 days cultured MGCs showed ovulatory genes mRNAs expression similar to the pre-ovulatory non-luteinized follicles. These FSH-stimulated cells responded to hCG stimulation in a pattern similar to the response of pre-ovulatory follicles. This novel model may provide a standardized research tool for delineation of the molecular processes occurring during the latter stages of follicular development in the human ovary.

Umbilical Cord Hernias Prenatal Diagnosis and Natural History

The purpose of this study was to describe the characteristics and outcomes of umbilical cord hernias diagnosed prenatally.

Progesterone antagonist, RU486, represses LHCGR expression and LH/hCG signaling in cultured luteinized human mural granulosa cells

Abstract Progesterone, the main steroid synthesized by the corpus luteum (CL), prepares the uterus for implantation, maintains the CL survival, and induces progesterone auto-secretion. However, the molecular mechanisms involving the progesterone auto-secretion pathways at the luteal phase are not fully understood, especially in humans. We aim to study the molecular mechanism of the progesterone pathway in human granulosa cells. Our model system consists of luteinized human-mural-granulosa-cells (hmGCs) obtained from follicles aspirated during in vitro fertilization (IVF) procedures. hmGCs were seeded in culture and were subjected to different hormonal treatments. mRNA levels were analyzed by quantitative real-time PCR (qRT-PCR). Progesterone levels were measured by enzyme immunoassay (EIA). We show that exposure of luteinized hmGCs to the progesterone receptor antagonist, RU486 (mifepristone), resulted in inhibition of LHCGR, LH/hCG target genes and progesterone secretion. Exposure of hmGCs to medium that was incubated with hmGCs for 4 d – conditioned medium (CM), which contain 150 ± 7.5 nM progesterone, resulted in induction of LHCGR and LH/hCG target genes, which was blocked by RU486. In addition, RU486 inhibited some of the progesterone biosynthesis pathway genes. Our results revealed a novel mechanism of the progesterone antagonist pathway in the luteal granulosa cells and emphasis the fundamental role of progesterone in the early luteal phase.

Perinatal outcome after fetal reduction from twin to singleton: to reduce or not to reduce?

OBJECTIVE To determine whether reduction of twin gestation to singleton pregnancy is associated with improved perinatal outcome. DESIGN A retrospective cohort study. SETTING Single tertiary care medical center. PATIENT(S) A cohort of 63 singleton pregnancies after reduction from dichorionic-diamniotic twins gestation and 62 dichorionic-diamniotic nonreduced twins. INTERVENTION(S) Fetal reduction between 11 and 14 weeks of gestation. MAIN OUTCOME MEASURE(S) Obstetric outcome. RESULT(S) The rates of preterm delivery at <34 weeks (1.6% in pregnancies after reduction vs. 11.7% in nonreduced twins) and at <37 weeks of gestation (9.5% vs. 56.7%) were significantly lower in patients whose pregnancies were reduced to singletons. The rates of miscarriage of one twin (0% vs. 4.8%) and early pregnancy loss before 24 weeks of gestation as well as the rates of gestational diabetes (11.1% vs. 10%), hypertensive diseases of pregnancy (6.3% vs. 15%), and intrauterine growth restriction (0% vs. 3.3%) were similar in both groups. CONCLUSION(S) Fetal reduction of twins to singleton is associated with a lower risk of prematurity and superior perinatal outcome compared with nonreduced twins. Therefore, the option of fetal reduction should be considered in certain cases of twin pregnancies, where the risk for adverse outcome seems exceptionally high.

Neurodevelopmental outcome of isolated ventriculomegaly: a prospective cohort study

Data regarding the neurodevelopmental outcome of children diagnosed in utero with isolated ventriculomegaly (IVM) are limited and principally founded on ultrasound‐based studies. Here, we endeavored to assess the outcome of such cases in a large‐scale, magnetic resonance imaging (MRI)‐based study.

Mode of delivery of twin gestation with very low birthweight: is vaginal delivery safe?

OBJECTIVE The purpose of this study was to determine whether planned vaginal delivery is associated with increased risk of perinatal death and morbidity in twin pregnancies that are complicated by a very low birthweight of the second twin. STUDY DESIGN We conducted a retrospective cohort study of twin pregnancies in which the second twins birthweight was ≤1500 g. One hundred ninety-three twin gestations met the study criteria; patients were classified into 2 groups according to the planned mode of delivery: (1) cesarean delivery (n = 142) and (2) vaginal delivery (n = 51). In the vaginal delivery group, 21 pairs were in cephalic-cephalic presentation at the time of delivery; 28 pairs were cephalic-noncephalic, and 2 pairs were noncephalic-noncephalic. Composite adverse neonatal outcome was defined as the presence of neonatal death, respiratory distress syndrome, sepsis, necrotizing enterocolitis, or intraventricular hemorrhage grade 3-4. RESULTS Trial of vaginal delivery was successful for both twins in 90.5% of cephalic-cephalic twins and 96.4% in cephalic-noncephalic twins. The rate of intraventricular hemorrhage was significantly higher in the vaginal delivery group (29.4% vs 8.5%, respectively; P = .013; adjusted odds ratio [OR], 3.65; 95% confidence interval [CI], 1.32-10.1). The increased risk of intraventricular hemorrhage in the vaginal delivery groups was evident in both twin A (17.6% vs 7.0%; P = .029) and twin B (15.7% vs 4.9%; P = .014); however, these differences were not significant after adjustment for possible confounders (twin A: adjusted OR, 1.79; 95% CI, 0.58-5.55; twin B: adjusted OR, 2.13; 95% CI, 0.63-7.25). In addition, subgroup analysis revealed that both cephalic-cephalic and cephalic-noncephalic twins who were delivered vaginally had increased risk for intraventricular hemorrhage. There were no significant differences between the cesarean and vaginal delivery groups in the rates of Apgar score <7 at 5 minutes, arterial cord pH <7.1, composite adverse neonatal outcome, and neonatal mortality rate. However, the rate of respiratory distress syndrome was significantly lower in the vaginal delivery group (66.7% vs 69%; P = .042; OR, 0.34; 95% CI, 0.12-0.96). CONCLUSION Vaginal delivery of very low birthweight twins is associated with an increased risk of intraventricular hemorrhage, regardless of presentation. Because of the small sample size and the retrospective cohort design, large prospective randomized studies are needed.

Safety of labor induction with prostaglandin E2 in grandmultiparous women

Objective: The aim of this study was to assess the safety of labor induction with vaginal prostaglandin E2 (PGE2) in grandmultiparous women. Methods: We conducted a retrospective cohort study of 1376 grandmultiparous women who underwent induction of labor with low dose PGE2. The primary outcome was uterine rupture and secondary outcomes included mode of delivery, postpartum hemorrhage and five minutes Apgar score. Results: One case was diagnosed with uterine rupture (0.07%). Vaginal delivery was achieved in 1329 (96.6%) patients, whereas 47 (3.4%) patients had emergent cesarean delivery. Five minutes Apgar score ≤7 was recorded in three cases (0.2%). There was no correlation between parity and cesarean delivery rate or low Apgar score. There were no significant differences between the grandmultiparous and great-grandmultiparous patients regarding cesarean delivery rate (3.1 vs. 5%, P = 0.12), operative vaginal delivery rate (2 vs. 2.3%, P = 0.74) or postpartum hemorrhage rate (0.8 vs. 1.1%, P = 0.6). Conclusions: Low dose PGE2 is a safe and efficient method for induction of labor in grandmultiparous and great-grandmultiparous women.