Search

It was suggested in the 1980s that long-term pituitary down-regulation by a gonadotrophin-releasing hormone agonist, termed the ultra-long protocol, inducing a hypo-oestrogenic state, might improve reproductive outcomes in women with endometriosis. Subsequently, international guidelines strongly supported the long-term pituitary down-regulation protocol in women with endometriosis based on a Cochrane review from 2006. The recently published European Society for Human Reproduction and Embryology guideline, based on the updated Cochrane review from 2019 and newer evidence, has reversed this recommendation. This paper explores the past and current evidence that led to these recommendations and calls for a consideration of refinement of the international guidelines to include additional factors and evaluate whether a paradigm shift is needed in the approach to endometriosis-related infertility. We believe that this can optimize evidence-based patient-centred care and benefit women worldwide and improve the design of future studies.

Objective In a previous study, we demonstrated that perinatal hypoxia leads to premature activation and initiation of growth in dormant follicles, leading to diminished ovarian reserve. Other reports have also established a connection between intrauterine deprivation, premature delivery, small for gestational age newborns, and decreased ovarian reserve, attributed to premature recruitment and depletion of ovarian follicles. However, it cannot be dismissed that an indirect mechanism, such as the release of stress-related hormones, may influence ovarian follicle recruitment under hypoxic conditions. Therefore, we conducted this study to determine whether hypoxic ovarian damage results from increased follicle growth and "burnout" or from increased apoptosis and whether this damage is dependent on age. Subjects Animal studies were carried out using adult 6-week-old (n = 8) and one-day-old newborn (n = 20) ICR (CD-1) female mice. The mice were sacrificed, and ovaries harvested and immediately cultured in Leibovitz media supplemented with L-Glutamine and 10% Fetal Bovine Serum. Ovarian tissue from both dams and pups was subjected to hypoxic (1% O2 and 99% N2)- or normoxic (21% O2 and 5% CO2) conditions and cultured for three hours at 37oC. The tissues were subsequently fixed in 4% formaldehyde for further processing and analyses. Follicular counts were conducted on H&E-stained sections, while immunohistochemistry was performed on sections that were stained with Ki-67 (cell proliferation marker), anti-Caspase 3 and anti-FOXO3A (apoptosis markers). Results Exposure to hypoxia resulted in a significant reduction in the proportion of primordial follicles out of the total follicular pool as compared to normoxia in both adult dams and newborn pups (3.17 ± 2.75% vs. 17.89 ± 4.4%; p = 0.004 and 40.59 ± 14.88% vs. 81.92 ± 31.56%, p = 0.001, respectively). This decrease was concomitant with an increase in the proportion of growing- primary and secondary follicles. Notably, the impact was strikingly more pronounced in adult dams than in newborn pups (6-fold vs. 2-fold, respectively). Ki67 staining revealed higher scores of cell proliferation in follicular granulosa cells after exposure to hypoxia than normoxia. However, Caspase 3 and Foxo3A staining did not show any differences in these markers of apoptosis in oocytes, granulosa cells, theca cells, or stromal cells when exposed to hypoxia versus normoxia. Conclusion Our study demonstrates that direct tissue hypoxia leads to the premature activation and initiation of growth in dormant follicles leading to diminished ovarian reserve. This effect is associated with increased granulosa cell proliferation without concomitant changes in apoptosis. Hypoxic damage is age dependent, with adult ovaries exhibiting a more pronounced susceptibility than newborn ovaries. Collectively, these findings support the possibility of follicular "burn out" as a potential mechanism responsible for hypoxia-induced loss of ovarian reserve.

Background Optimal intensive luteal support in frozen embryo transfer (FET) cycles in which a corpus luteum is not present has not been determined and several treatment regimens are currently practiced. Previous reports suggest that vaginal progesterone alone is probably insufficient. The use of intramuscular progesterone is considered efficacious but much less tolerable. The objective of this study was to compare a more tolerable regimen using micronized vaginal progesterone (MVP) and oral dydrogesterone versus MVP and intramuscular progesterone (IM) for luteal support. Methods A retrospective study of patients undergoing single blastocyst FET in a tertiary center IVF unit. All patients (n = 100) received oral estradiol in the follicular phase and supplemented with MVP for luteal phase support. One group (n = 50) received additional dydrogesterone (10 mg TID) and the other (n = 50) intramuscular progesterone (100 mg every 3 days). Results The two groups were similar with regard to age; BMI; infertility etiology and duration; and basal hormonal levels. Biochemical pregnancy rates (34% vs 42%, p = 0.54) and clinical pregnancy rates (26% vs. 36%, p = 0.39) were lower in the MVP + oral dydrogesterone group as compared to the MVP + IM progesterone group. Nevertheless, this difference was not statistically significant. Abortion rates were similar (2%) and there were no ectopic pregnancies in both groups. Conclusion This retrospective cohort study demonstrates noninferiority of MVP and oral dydrogesterone as compared to MVP and intramuscular progesterone for luteal support in FET cycles. The better profile of adverse effects and tolerability makes it a suitable protocol for FET.

The human ovary contains 6-million follicles during the 20th week of embryonic development and 1 million at birth. Girls born at small for gestational age weight demonstrate higher FSH levels during infancy, an earlier onset of puberty, and menarche. In light of these observations, we hypothesized that exposure to hypoxia at the early neonatal period might impact the primordial follicular pool and lead to premature depletion of ovarian reserve.Ovarian development in the rat model at days 1-5 postpartum reflects its human counterpart in the late perinatal period. We exposed newborn rat pups (The percentage of primordial follicles out of total follicles in ovaries of pups exposed to hypoxia was lower compared to control (76 ± 8.2% and 90.33 ± 6.3% respectively,We provide evidence for the first time that perinatal hypoxia causes premature activation and growth initiation of dormant follicles. These changes were associated with decreased stromal cell proliferation, suggesting hypoxia-induced impairment of the support cell pool as a possible mechanism for accelerated follicular activation.

Maternal natural vaginal progesterone (nVP) administration has been shown to reduce the risk of preterm birth (PTB). The largest randomized trial of nVP for PTB (OPPTIMUM) noted a sonographic reduction in neonatal brain injury following nVP treatment. We investigated the neuroinflammatory protective effect of maternal nVP in a mouse model for maternal inflammation. Pregnant mice (n = 24) were randomized to nVP (1 mg/day) or vehicle from days 13–16 of gestation. At days 15 and 16, lipopolysaccharide (30 μg) or saline were administered. Mice were sacrificed 4 h following the last injection. Fetal brains and placentas were collected. Levels of NF-κB, nNOS, IL-6, and TNFα were determined by Western blot. Maternal lipopolysaccharide significantly increased fetal brain levels of IL-6 (0.33 ± 0.02 vs. 0.11 ± 0.01 u), TNFα (0.3 ± 0.02 vs. 0.10 ± 0.01 u), NF-κB (0.32 ± 0.01 vs. 0.17 ± 0.01 u), and nNOS (0.24 ± 0.04 vs. 0.08 ± 0.01 u), and reduced the total glutathione levels (0.014 ± 0.001 vs. 0.026 ± 0.001 pmol/μl; p

We evaluated the effect of in vitro fertilization (IVF) on sexual function in men, particularly for erectile dysfunction.A prospective case-control study at a tertiary medical center. The study group comprised men of infertile couples that required IVF to conceive. The control group comprised men of couples who conceived spontaneously. The effects of IVF on sexual and erectile function were assessed based on the International Index of Erectile Function (IIEF-15) and the Self-Esteem and Relationship (SEAR) questionnaires. Participants were followed up to 1 year postpartum.Compared to the control group (378), for the IVF group (356), mean IIEF-15 scores were significantly lower: prior to pregnancy (31.7±4.5 vs 64.4±7.2, p0.0001), at mid-pregnancy (37.3±5.1 vs 66.4±5.5, p0.0001) and up to one year postpartum (42.3±4.9 vs 68.6±4.3, p0.0001). Compared to the control group, in the IVF group, mean SEAR scores were significantly lower at these 3 respective time points (29.9±6.3 vs 66.5±8.3; 34.1±5.8 vs 66.9±7.2; and 40.9±6.7 vs 67.3±5.6; p0.0001). At the 3 time points, for the IVF compared to the control group, the median monthly sexual intercourse rate was lower; and both the use of phosphodiesterase-5 inhibitor and psychologist/sexologist care were higher.The prevalence of erectile dysfunction among men participating in IVF in order to conceive is significantly higher compared to couples that conceived spontaneously, thus leading to an extremely high rate of phosphodiesterase-5 inhibitor use.

Liver regeneration depends on sequential activation of pathways and cells involving the remaining organ in recovery of mass. Proliferation of parenchyma is dependent on angiogenesis. Understanding liver regeneration-associated neovascularization may be useful for development of clinical interventions. Myeloid-derived suppressor cells (MDSCs) promote tumor angiogenesis and play a role in developmental processes that necessitate rapid vascularization. We therefore hypothesized that the MDSCs could play a role in liver regeneration. Following partial hepatectomy, MDSCs were enriched within regenerating livers, and their depletion led to increased liver injury and postoperative mortality, reduced liver weights, decreased hepatic vascularization, reduced hepatocyte hypertrophy and proliferation, and aberrant liver function. Gene expression profiling of regenerating liver-derived MDSCs demonstrated a large-scale transcriptional response involving key pathways related to angiogenesis. Functionally, enhanced reactive oxygen species production and angiogenic capacities of regenerating liver-derived MDSCs were confirmed. A comparative analysis revealed that the transcriptional response of MDSCs during liver regeneration resembled that of peripheral blood MDSCs during progression of abdominal tumors, suggesting a common MDSC gene expression profile promoting angiogenesis. In summary, our study shows that MDSCs contribute to early stages of liver regeneration possibly by exerting proangiogenic functions using a unique transcriptional program.-Nachmany, I., Bogoch, Y., Sivan, A., Amar, O., Bondar, E., Zohar, N., Yakubovsky, O., Fainaru, O., Klausner, J. M., Pencovich, N. CD11b+Ly6G+ myeloid-derived suppressor cells promote liver regeneration in a murine model of major hepatectomy.

Abstract Progesterone has been shown to regulate immunity during pregnancy, and progesterone administration may reduce inflammation-induced preterm labor. We sought to determine the maternal brain immune response to LPS-induced inflammation in pregnant and non-pregnant mice and whether additional progesterone supplementation attenuates this response. Pregnant (P: n = 9) and non-pregnant mice (NP: n = 9) were randomized to pretreatment with vaginal progesterone/carrier (Replens), daily from days 13 to 16. On days 15 and 16, LPS/saline was administered by intraperitoneal injection (Replens + saline n = 3; Replens + LPS n = 3; progesterone + LPS n = 3). Mice were sacrificed on day 16 and maternal serum analyzed for IL-6 levels and brains analyzed for nNOS, NF-kB, IL-6 protein levels and for immature myeloid cells (IMCs) and microglial activity. LPS significantly increased brain nNOS, NF-kB, and IL-6 in both NP and P mice, with significantly greater responses in P mice. In both NP and P groups, progesterone significantly attenuated LPS-induced increase of nNOS and NF-kB, however with no effect on serum IL-6. In the NP brains, LPS significantly increased IMC population and progesterone reduced the IMC phenotype to levels similar to controls. In P mice, neither LPS nor LPS + progesterone altered the brain IMC population. LPS significantly increased the microglial activity in both NP and P groups, which was attenuated by progesterone. Progesterone attenuates brain inflammatory response to LPS in both NP and P mice although it has no effect on systemic inflammation. In NP mice, progesterone attenuated the increase in brain IMC following LPS administration. Our results suggest that endogenous progesterone during pregnancy may protect the brain from LPS-induced inflammation.

BACKGROUND Necrotizing enterocolitis (NEC) is a devastating gastrointestinal disease of neonates, especially premature neonates. To date, there is no prophylactic treatment against NEC, except breast milk and slow increase in enteral feeding, and there is no antenatal prophylaxis. AIMS To assess possible protective effects of antenatal N-Acetyl Cysteine (NAC) against the intestinal pathophysiological changes associated with NEC in a rat model of NEC and against its associated mortality. METHODS Newborn Sprague-Dawley rats were divided into 5 groups: control (n = 33); NEC (n = 32)-subjected to hypoxia and formula feeding for 4 days to induce NEC; NEC-NAC (n = 34)-with induced NEC and concomitant postnatal NAC administration; NAC-NEC (n = 33)-born to dams treated with NAC for the last 3 days of pregnancy starting at gestational age of 18 days, and then subjected to induced NEC after birth; NAC-NEC-NAC (n = 36)-subjected to induced NEC with both prenatal and postnatal NAC treatment. At day of life 5, weight and survival of pups in the different groups were examined, and pups were euthanized. Ileal TNF-α, IL-6, IL-1β, IL-10, NFkB p65, iNOS and cleaved caspase 3 protein levels (western blot) and mRNA expression (RT-PCR) were compared between groups. RESULTS Pup mortality was significantly reduced in the NAC-NEC-NAC group compared to NEC (11% vs. 34%, P

New drug approval requires a new drug to undergo rigorous clinical trials to determine its efficacy and safety. A drug is approved only for the population on which it was tested, i.e. those who meet the inclusion criteria of the trial. The aim of this study was to determine what percentage of 'real life' patients in our clinic meet the inclusion and exclusion criteria used in large-scale clinical trials required for drug registration in the field of assisted reproduction. All 265 consecutive patients with pertinent data treated in a tertiary centre IVF Unit during 2015 were surveyed. Their demographic and clinical parameters were compared with inclusion and exclusion criteria used in nine major clinical trials. Only 97 out of 265 (37%) patients met the consensus inclusion criteria as defined by the nine clinical trials. The number of oocytes retrieved was 9.10 ± 5.34 in the patients that met the inclusion criteria (n = 97) versus 6.90 ± 5.23 (P = 0.00122) in those that did not (n = 168). Most 'real life' patients who come for treatment at a tertiary IVF centre do not meet the consensus of inclusion and exclusion criteria used for major clinical trials.

Liver regeneration depends on sequential activation of pathways and cells involving the remaining organ in recovery of mass. Proliferation of parenchyma is dependent on angiogenesis. Understanding liver regeneration-associated neovascularization may be useful for development of clinical interventions. Myeloid-derived suppressor cells (MDSCs) promote tumor angiogenesis and play a role in developmental processes that necessitate rapid vascularization. We therefore hypothesized that the MDSCs could play a role in liver regeneration. Following partial hepatectomy, MDSCs were enriched within regenerating livers, and their depletion led to increased liver injury and postoperative mortality, reduced liver weights, decreased hepatic vascularization, reduced hepatocyte hypertrophy and proliferation, and aberrant liver function. Gene expression profiling of regenerating liver-derived MDSCs demonstrated a large-scale transcriptional response involving key pathways related to angiogenesis. Functionally, enhanced reactive oxygen species production and angiogenic capacities of regenerating liver-derived MDSCs were confirmed. A comparative analysis revealed that the transcriptional response of MDSCs during liver regeneration resembled that of peripheral blood MDSCs during progression of abdominal tumors, suggesting a common MDSC gene expression profile promoting angiogenesis. In summary, our study shows that MDSCs contribute to early stages of liver regeneration possibly by exerting proangiogenic functions using a unique transcriptional program.-Nachmany, I., Bogoch, Y., Sivan, A., Amar, O., Bondar, E., Zohar, N., Yakubovsky, O., Fainaru, O., Klausner, J. M., Pencovich, N. CD11b

Final oocyte maturation is a pivotal step in in vitro fertilization, traditionally achieved by a single bolus of human chorionic gonadotropin (hCG) administered 36 h before oocyte retrieval. The introduction of GnRH antagonists paved the way to trigger final oocyte maturation with GnRH agonists. The endogenous LH and FSH surges following GnRH agonist triggering, although shorter than their natural counterparts, are sufficient to yield mature oocytes. In most patients, GnRH agonist triggering is followed by a complete and irreversible luteolysis, hence almost complete prevention of ovarian hyperstimulation syndrome (OHSS), even following extreme ovarian responses. “Freeze-all” and IVF “segmentation” may serve to achieve OHSS-free environment. If “fresh” transfer is desirable, careful luteal-phase support is mandatory, for which numerous protocols have been suggested. The use of these protocols must be carefully tailored against the full scope of individual clinical parameters.

Myeloid derived suppressor cells (MDSCs) play key roles in cancer development. Accumulation of peripheral-blood MDSCs (PB-MDSCs) corresponds to the progression of various cancers, but provides only a crude indicator. We aimed toward identifying changes in the transcriptional profile of PB-MDSCs in response to tumor growth. CT26 colon cancer cells and B16 melanoma cells (106) were inoculated into peritoneal cavities of BALB/c mice and subcutaneously to C57-black mice, respectively. The circulating levels and global transcriptional patterns of PB CD11b+Ly6g+ MDSCs were assessed in control mice, and 4, 8, and 11 days following tumor cell inoculation. Although a significant accumulation of PB-MDSCs was demonstrated only 11 days following tumor induction, a pronounced transcriptional response was identified already on day 4 while the tumor was ~1 mm in size. Further transcriptional changes correlated with different stages of tumor growth. Key MDSC genes and canonical signaling pathways were activated along tumor progression. This phenomenon was demonstrated in both cancer models, and a consensus set of 817 genes, involved in myeloid cell recruitment and angiogenesis, was identified. The data suggest that the transcriptional signatures of PB-MDSC may serve as markers for tumor progression, as well as providing potential targets for future therapies.

Introduction immature-myeloid cells (IMCs) are proangiogenic bone marrow (BM)-derived cells that normally differentiate into inflammatory cells such as neutrophils, monocytes and dendritic cells (DCs). We characterized placental IMCs comparing their gene expression and subpopulations to tumor IMCs, and tested our hypothesis that progesterone that inhibits preterm labor, may affect their abundance and differentiation. Methods differences between IMC-subpopulations in subcutaneous tumors versus placentas in C57BL/6 or ICR (CD-1) mice were analyzed by flow cytometry and gene expression was detected by microarrays. BM- and placental cells were incubated with or without progesterone and IMC subpopulations were analyzed. For preterm labor induction pregnant mice pretreated or not with progesterone were or were not treated with Lipopolysaccharide (LPS). Results we detected enrichment of granulocytic-IMCs in placentas compared to tumors, paralleled by a decrease in monocytic-IMCs. mRNA expression of placenta- versus tumor IMCs revealed profound transcriptional alterations. Progesterone treated BM-CD11b+ cells ex-vivo induced enrichment of granulocytic-IMCs and a decrease in monocytic-IMCs and DCs. LPS treatment in-vivo led to an increase in BM-IMCs in both progesterone pretreated or non-pretreated mice. In the placenta LPS decreased the IMC population while progesterone led to complete abrogation of this effect. Discussion placental IMCs differ from tumor-IMCs in both subpopulations and gene expression. Progesterone enhances the proliferation of placenta-specific granulocytic IMCs ex-vivo and LPS induced labor is accompanied by a decrease in placental IMCs only in progesterone non-pretreated mice. We thus speculate that the protective effect of progesterone in preventing preterm labor may be explained at least in part by this specific anti-inflammatory effect.

Although escape from tumor dormancy has long been recognized as an important problem in the treatment of cancer, the molecular and cellular regulators underlying this transition remain poorly understood. The inability of the cancer cells to induce a complete and successful process of angiogenesis can result in tumor dormancy. In this case, the acquisition of sufficient angiogenic potential will result in the escape from indolence and in the initiation of tumor mass expansion. This stage in disease progression is known as the angiogenic switch. It is now becoming clear that the induction of the angiogenic switch is controlled by dynamic and complex biological processes involving the cancer cells, the associated stromal microenvironment and distant normal host cells, mostly from the bone marrow. Indeed, intricate tumor-host interactions are increasingly recognized as critical features of cancer. In particular, infiltrating cells of the immune system are crucial constituents of tumors and an important source of the growth stimulatory signals to the tumor cells. Tumor cells are surrounded by stromal cells, such as fibroblasts, lymphocytes, neutrophils, macrophages and mast cells, which communicate via a complex network of intercellular signaling pathways, mediated by surface adhesion molecules, cytokines and their receptors. However, the possible roles of these cells and molecules in the maintenance of micro-tumors in an occult state and in the induction of exit from the dormant state are not fully elucidated. In this review, we summarize recent findings and the current understanding of the role of bone marrow-derived cells, their recruitment into tumors and their interactive crosstalk with tumor cells, in leading to either the maintenance of, or exit from, tumor dormancy. Understanding the mechanisms of tumor growth and metastatic recurrence after periods of indolence is crucial for improving early detection, as well as increasing the cure rate for cancer patients.

Objective Ovarian hyperstimulation syndrome is associated with increased angiogenesis and vascular leakage. Immature myeloid cells (IMCs) and dendritic cells have been shown to be actively involved in angiogenesis in several disease models in mice and humans. Nevertheless, little is known about the role of these cells in the ovary. As such, this study sought to determine whether alterations in these ovarian myeloid cell populations are associated with gonadotropin stimulation in a mouse model. Study design Four-week-old pre-pubertal C57Bl/6 female mice were allocated into three groups: high-dose stimulation ( n = 4; pregnant mare serum gonadotropins (PMSG) 20 U for 2 days), low-dose stimulation ( n = 5; PMSG 5 U for 1 day) and sham-treated controls ( n = 4). Human chorionic gonadotropin 5 U was injected on Day 3, and the mice were killed on Day 5. Ovaries were analysed by flow cytometry, confocal microscopy and quantitative polymerase chain reaction. Results Gonadotropin stimulation increased the proportion of CD11b + Gr1 + IMCs among the ovarian myeloid cells: 22.6 ± 8.1% (high dose), 7.2 ± 1.6% (low dose) and 4.1 ± 0.3% (control) ( p = 0.02). Conversely, gonadotropin stimulation decreased the proportion of ovarian CD11c + MHCII + dendritic cells: 15.1 ± 1.9% (high dose), 20.7 ± 4.8% (low dose) and 27.3 ± 8.2% (control) ( p = 0.02). IMCs, unlike dendritic cells, were localized adjacent to PECAM1 + endothelial cells. Finally, gonadotropin stimulation was associated with increased expression of S100A8, S100A9, Vcan and Dmbt1, and decreased expression of MMP12. Conclusions Gonadotropin stimulation is associated with proangiogenic myeloid cell alterations, reflected by a dose-dependent increase in ovarian IMCs and a parallel decrease in dendritic cells. Recruited IMCs localize strategically at sites of angiogenesis. These changes are associated with differential expression of key proangiogenic genes.

Objective To investigate whether "proangiogenic" CD56 + CD16 − natural killer (NK) cells, which accumulate in follicular fluid (FF) of patients with a good response to ovarian stimulation, are also present in earlier stages of follicular development. Design Observational study. Setting Academic in vitro fertilization (IVF) unit. Patient(s) Patients of similar age and ovarian reserve, undergoing in vitro maturation (IVM; n=10) or IVF (n = 22) cycles. Intervention(s) None. Main Outcome Measure(s) FF was collected from the first follicle aspirated in each ovary, and flow cytometry was used to define CD56 + CD16 − "proangiogenic" or CD56 + CD16 + "cytotoxic" NK cells. Result(s) FF derived from antral follicles of patients undergoing IVM (maximum diameter 10 mm) showed a slightly higher abundance of "proangiogenic" NK cells compared with FF from preovulatory mature follicles (>18 mm) of patients undergoing IVF (5.4 ± 1.3% vs. 3.0 ± 1.1% of CD45 + CD3 − cells). Importantly, antral FF contained a significantly higher concentration of "cytotoxic" NK cells (11.4 ± 2.3% vs. 3.7 ± 0.9% of CD45 + CD3 − cells) compared with FF from mature follicles. Conclusion(s) "Proangiogenic" NK cells accumulate in ovarian follicles from as early as the antral follicular stage. Maturation of follicles is accompanied by a decrease in the population of "cytotoxic" NK cells that may have deleterious effects on follicular maturation.

CD56(bright)CD16(-) natural killer (NK) cells that express CXCR3 accumulate in the follicular fluid of patients with a good response to ovarian stimulation. It is conceivable that granulosa cells of good responders that express the chemokine CXCL10 recruit these NK cells to support follicular angiogenesis.

Dendritic cells (DCs)—immunomodulatory cells that initiate adaptive immune responses—have recently been shown to exert proangiogenic effects when infiltrating the tumor microenvironment. As tumors that escape immune surveillance inhibit DC maturation, we explored whether maturation status determines their ability to promote angiogenesis and whether angiogenesis depends on the presence of DCs. Using mouse xenograft models of human tumors, we show that fast-growing “angiogenic” tumors are infiltrated by a more immature DC population than respective dormant avascular tumors. Accordingly, supplementation of immature DCs, but not mature DCs, enhanced tumor growth. When DCs were mixed with Matrigel and injected subcutaneously into mice, only immature DCs promoted the ingrowth of patent blood vessels. Notably, depletion of DCs in a transgenic mouse model that allows for their conditional ablation completely abrogated basic fibroblast growth factor-induced angiogenesis in Matrigel plugs, and significantly inhibited tumor growth in these mice. Because immature DCs actively promote angiogenesis and tumor growth, whereas DC maturation or ablation suppresses this response, we conclude that angiogenesis is dependent on the presence of immature DCs. Thus, cancer immunotherapies that promote DC maturation may act by both augmenting the host immune response to the tumor and by suppressing tumor angiogenesis.—Fainaru, O., Almog, N., Yung, C. W., Nakai, K., Montoya-Zavala, M., Abollahi, A., D’Amato, R., Ingber, D. E. Tumor growth and angiogenesis are dependent on the presence of immature dendritic cells.

The endothelium lining blood vessels serves as a barrier against vascular hyperpermeability, and its maintenance is critical to organ health. Inflammatory mediators evoke tissue edema by disrupting the expression of membrane junctional proteins, which mediate binding between endothelial cell membranes. Endothelial cell-cell junctions form a diffusion barrier between the intravascular and interstitial space. To prevent the morbidity and mortality caused by exaggerated vascular permeability associated with pathological states (e.g., inflammatory and hypersensitivity disorders, pulmonary edema, traumatic lung injury, cerebral edema resulting from stroke, and others), it is important to develop therapeutic approaches to stabilize these interendothelial junctions. Vascular endothelial growth factor (VEGF), a potent proangiogenic cytokine, was first described as vascular permeability factor (VPF). Doxycycline, a tetracycline derivative, has been shown to inhibit angiogenesis in both humans and animal models. We now report that oral doxycycline prevents VPF/VEGF-induced vascular permeability, interleukin-2-induced pulmonary edema, and delayed-type hypersensitivity (DTH) in mice. Remarkably, doxycycline also inhibits tumor growth and tumor-associated vascular hyperpermeability. Finally, we show that doxycycline targets the adherens junction in vascular endothelial cells by inducing the total amount of VE-cadherin expression while decreasing the degree of its phosphorylation. The potential of doxycyline as a therapeutic inhibitor of vascular hyperpermeability in human clinical conditions is promising and warrants further studies.

Targeting angiogenesis, the formation of blood vessels, is an important modality for cancer therapy. TNP-470, a fumagillin analog, is among the most potent and broad-spectrum angiogenesis inhibitors. However, a major clinical limitation is its poor oral availability and short half-life, necessitating frequent, continuous parenteral administration. We have addressed these issues and report an oral formulation of TNP-470, named Lodamin. TNP-470 was conjugated to monomethoxy-polyethylene glycol–polylactic acid to form nanopolymeric micelles. This conjugate can be absorbed by the intestine and selectively accumulates in tumors. Lodamin significantly inhibits tumor growth, without causing neurological impairment in tumor-bearing mice. Using the oral route of administration, it first reaches the liver, making it especially efficient in preventing the development of liver metastasis in mice. We show that Lodamin is an oral nontoxic antiangiogenic drug that can be chronically administered for cancer therapy or metastasis prevention.

Endometriosis affects 10-15% of women and is associated with pelvic pain and infertility. Angiogenesis plays an essential role in its pathogenesis. Dendritic cells (DCs) were recently implicated in supporting tumor angiogenesis. As both tumors and endometriosis lesions depend on angiogenesis, we investigated the possibility that DCs may also play a role in endometriosis. We induced endometriosis in 8-wk-old female C57BL/6 mice by implantation of autologous endometrium into the peritoneal cavity. We observed an abundance of CD11c(+) DCs infiltrating sites of angiogenesis in endometriosis lesions. We noticed a similar pattern of infiltrating DCs at sites of angiogenesis in the peritoneal Lewis lung carcinoma tumor model. These DCs were immature (major histocompatability complex class II(low)) and expressed vascular endothelial growth factor receptor 2. Peritoneal implanted bone marrow-derived DCs (BMDCs) incorporated into both endometriosis lesions and into B16 melanoma tumors and enhanced their growth at 8 days compared with controls (5.1+/-2.5 vs. 1.5+/-0.5 mm(2), n=4 and 4, P

Objective In our previous studies we have shown that the process of term labor is associated with oxidative stress, as indicated by increased susceptibility of maternal serum lipids to copper induced peroxidation. In order to continue evaluating the role of oxidative stress in the labor process, we next tested whether term premature rupture of the membranes (PROM) is also associated with increased susceptibility of maternal serum lipids to copper induced peroxidation. Design A controlled prospective study. Setting Tertiary care centre. Population 31 healthy women with term PROM and 19 healthy pregnant women with intact membranes. The women were matched for maternal and gestational age. Methods Venous blood was drawn from the women (up to 6h after rupture of the membranes and prior to labor in the PROM group), and the kinetics of copper-induced oxidation of serum lipids ex vivo were monitored spectroscopically at 37°C by continuous recording of absorbance at 245nm. Results The lag phase, reflecting resistance of serum lipids to oxidation, was similar in the PROM group when compared to the control group (43.7±3.2 versus 41.9±1.6min, P =0.61). However, the maximal rate of oxidation ( V max ) and the maximal accumulation of absorbing products (OD max ) were shorter in the PROM group when compared to the control group (5.14±0.26 versus 6.29±0.4010 −3 OD 245 nm/min, P =0.016; 0.61±0.03 versus 0.71±0.04 OD 245 nm, P =0.07). Conclusion As opposed to term labor, term PROM is not associated with increased maternal systemic oxidative stress when compared to normal pregnant women. The role for oxidative stress in preterm PROM warrants further studies.

Regulation of gene expression by tissue-specific transcription factors involves both turning on and turning off transcription of target genes. Runx3, a runt-domain transcription factor, regulates cell-intrinsic functions by activating and repressing gene expression in sensory neurons, dendritic cells (DC), and T cells. To investigate the mechanism of Runx3-mediated repression in an in vivo context, we generated mice expressing a mutant Runx3 lacking the C-terminal VWRPY, a motif required for Runx3 interaction with the corepressor Groucho/transducin-like Enhancer-of-split (TLE). In contrast with Runx3 −/− mice, which displayed ataxia due to the death of dorsal root ganglia TrkC neurons, Runx3 VWRPY−/− mice were not ataxic and had intact dorsal root ganglia neurons, indicating that ability of Runx3 to tether Groucho/TLE is not essential for neurogenesis. In the DC compartment, the mutant protein Runx3 VWRPY− promoted normally developed skin Langerhans cells but failed to restrain DC spontaneous maturation, indicating that this latter process involves Runx3-mediated repression through recruitment of Groucho/TLE. Moreover, in CD8 + thymocytes, Runx3 VWRPY− up-regulated α E /CD103-like WT Runx3, whereas unlike wild type, it failed to repress α E /CD103 in CD8 + splenocytes. Thus, in CD8-lineage T cells, Runx3 regulates α E /CD103 in opposing regulatory modes and recruits Groucho/TLE to facilitate the transition from activation to repression. Runx3 VWRPY− also failed to mediate the epigenetic silencing of CD4 gene in CD8 + T cells, but normally regulated other pan-CD8 + T cell genes. These data provide evidence for the requirement of Groucho/TLE for Runx3-mediated epigenetic silencing of CD4 and pertain to the mechanism through which other Runx3-regulated genes are epigenetically silenced.

Objective: Our purpose was to determine whether intracervical placement of laminaria stalks would improve the effectiveness of inducing termination of pregnancies in the second trimester by intra-amniotic injection of prostaglandin F2alpha. Study design: This is a prospective randomized clinical trial conducted at Gynecology Department, Lis Maternity Hospital. Fifty women admitted for indicated second trimester pregnancy termination were randomly assigned to receive either intracervical placement of laminaria (n=25) or not (n=25), with concurrent use of intra-amniotic injection of prostaglandin F2alpha and concentrated oxytocin. The outcome measures were: time from induction to delivery and the side effects and complications of treatment. Results: There was no significant difference in the mean interval from induction to delivery in the laminaria and no laminaria groups, being 20±1.3 (8–30) h versus 19±1.0 (8–26) h , respectively. In addition, this interval was not different in subgroups of primiparas or multiparas. Other outcome measures such as retained placenta (4% in both groups), post-partum hemorrhage (4% in both groups) gastro-intestinal side effects, fever, and use of analgesia were not significantly different between both groups. Conclusion: We found no additional value in placing laminaria stalks when using intra-amniotic injection of prostaglandin F2alpha followed by concentrated oxytocin infusion for second trimester termination of pregnancy.

RUNX transcription factors are key regulators of lineage-specific gene expression and might be involved in autoimmune diseases. Runx3 plays a role during the development of sensory neurons and T cells and regulates transforming growth factor β (TGF-β) signaling in dendritic cells. Here, we report that at 4 weeks of age, Runx3 knockout (KO) mice spontaneously develop inflammatory bowel disease (IBD) characterized by leukocyte infiltration, mucosal hyperplasia, formation of lymphoid clusters, and increased production of IgA. Additionally, at a considerably older age (8 months), the KO mice also develop progressive hyperplasia of the gastric mucosa associated with disturbed epithelial differentiation and cellular hyaline degeneration. Analysis of cytokines in the colonic mucosa of Runx3 KO mice revealed a mixed T helper 1/T helper 2 response. By using immunohistochemistry and RNAin situhybridization, Runx3 expression in the gastrointestinal tract is detected in lymphoid and myeloid populations but not in the epithelium. The data indicate that loss of leukocytic cell-autonomous function of Runx3 results in IBD and gastric lesion in the KO mice. IBD in humans is viewed as a complex genetic disorder. Several susceptibility loci were identified on different human chromosomes including the chromosomal region 1p36 whereRUNX3resides. It is thus tempting to speculate that mutations inRUNX3may constitute an IBD risk factor in humans.

Runx3 transcription factor regulates cell lineage decisions in thymopoiesis and neurogenesis. Here we report that Runx3 knockout (KO) mice develop spontaneous eosinophilic lung inflammation associated with airway remodeling and mucus hypersecretion. Runx3 is specifically expressed in mature dendritic cells (DC) and mediates their response to TGF-beta. In the absence of Runx3, DC become insensitive to TGF-beta-induced maturation inhibition, and TGF-beta-dependent Langerhans cell development is impaired. Maturation of Runx3 KO DC is accelerated and accompanied by increased efficacy to stimulate T cells and aberrant expression of beta2-integrins. Lung alveoli of Runx3 KO mice accumulate DC characteristic of allergic airway inflammation. Taken together, abnormalities in DC function and subset distribution may constitute the primary immune system defect, which leads to the eosinophilic lung inflammation in Runx3 KO mice. These data may help elucidate the molecular mechanisms underlying the pathogenesis of allergic airway inflammation in humans.

Background. Several reports suggest preeclampsia to be associated with oxidative stress. In view of potential experimental artifacts in these studies, we tested the effect of preeclampsia on the oxidizibility of maternal serum lipids, using an optimized ex vivo method. Methods. This prospective study included 28 pregnant women with preeclampsia and 28 women matched for maternal and gestational age with normal pregnancies. Venous blood was drawn from the consenting women. Serum levels of lipid peroxidation products and the kinetics of copper-induced oxidation ex vivo were monitored spectroscopically by continuous recording of absorbance at 245 nm. Results. The initial optical density (OD) at 245 nm, attributed to preformed products of lipid peroxidation, was higher in the women with preeclampsia than in the controls (1.26 ± 0.02 vs. 1.17 ± 0.02 OD units; p = 0.01). The lag phase preceding oxidation, reflecting resistance of serum lipids to oxidation, was significantly shorter in the preeclampsia group than...

This study aimed to evaluate the clinical and economic implications of integrating human papilloma virus (HPV) load testing into the follow-up and management protocol of women postconisation for high-grade cervical intraepithelial neoplasia (CIN2-3). We evaluated 130 suitable women: 63 were screened biannually by Pap smears ('conventional approach') and 67 also had HPV-load testing ('HPV approach'). More stringent criteria for undergoing colposcopy or reconisation were observed by the former group compared to the latter. Both approaches were analysed for cost effectiveness. There were 33 out of 67 (49.2%) colposcopic referrals and 24 out of 67 (35.8%) reconisation/hysterectomies with the 'conventional approach' compared to 9 out of 63 (14.2%) and 7 out of 63 (11.1%) with the 'HPV approach'. Cervical intraepithelial neoplasia 2-3 residual disease was detected in 7 out of 67 (10.5%) and 7 out of 63 (11.1%) women. The 'conventional approach' had more negative colposcopic biopsies and more negative reconisation/hysterectomy histologies than the 'HPV approach'. The respective cost per detection of one case of residual disease was US$3573 and US$3485. The 'HPV approach' required fewer colposcopic and reconisation procedures to detect one case of residual CIN2-3. Its higher positive predictive value than that of cytology provided a significant decrease in false positive rates and a reduction of US$88 per detected case.

The RUNX transcription factors are important regulators of lineage-specific gene expression. RUNX are bifunctional, acting both as activators and repressors of tissue-specific target genes. Recently, we have demonstrated that Runx3 is a neurogenic transcription factor, which regulates development and survival of proprioceptive neurons in dorsal root ganglia. Here we report that Runx3 and Runx1 are highly expressed in thymic medulla and cortex, respectively, and function in development of CD8 T cells during thymopoiesis. Runx3-deficient (Runx3 KO) mice display abnormalities in CD4 expression during lineage decisions and impairment of CD8 T cell maturation in the thymus. A large proportion of Runx3 KO peripheral CD8 T cells also expressed CD4, and in contrast to wild-type, their proliferation ability was largely reduced. In addition, thein vitrocytotoxic activity of alloimmunized peritoneal exudate lymphocytes was significantly lower in Runx3 KO compared with WT mice. In a compound mutant mouse, null for Runx3 and heterozygous for Runx1 (Runx3-/-;Runx1+/-), all peripheral CD8 T cells also expressed CD4, resulting in a complete lack of single-positive CD8+T cells in the spleen. The results provide information on the role of Runx3 and Runx1 in thymopoiesis and suggest that both act as transcriptional repressors ofCD4expression during T cell lineage decisions.

The upper limit of the fetal atrial width in normal fetuses is debated. This was a prospective cohort evaluation of the lateral ventricular width with special regard to the upper limit of its size.Measurements of fetal atrial ventricular size were obtained by abdominal ultrasonography in 427 male and female fetuses between 20 and 40 weeks' gestation of normal singleton pregnancies. In addition, reanalysis of previous data (8 studies) and the current data was performed to produce a pooled mean and SD.The mean ventricular width +/- SD was 6.2 +/- 1.2 mm. The ventricular width did not show significant modification throughout gestation. Reanalysis of the current and previous studies (8216 cases) yielded a pooled mean of 6.4 +/- 1.2.According to the current and previous studies, the upper cutoff of fetal ventricular atrium width should be 10 mm. This cutoff represents a range of approximately 3 SDs above the pooled mean, corresponding to a 99.74% confidence interval.

Objective: To investigate placental apoptosis in discordant dichorial twins. Methods: Placental samples were obtained from 7 third-trimester suitable twins. Discordancy was defined as a >25 per cent difference in newborn birth weight. Light microscopy using hematoxylin and eosin (H&E)-stained paraffin slides and terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end-labelling (TUNEL) methods were used to confirm the incidence of apoptosis. Investigators were blinded to pregnancy outcome. Results: Both methods revealed that the incidence of apoptosis in the placentas of the smaller fetuses was significantly higher than in placentas of the larger fetuses. The incidence of TUNEL-positive cells in the former was 1.4±0.26 per cent: this was significantly higher than the incidence of apoptosis in the placental specimens of the latter (0.9±0.07 per cent, P 0.02 Wilcoxon rank test). The same results were obtained with H&E: the incidence of apoptosis detected in placentas from the former was 1.07±0.1 per cent compared to 0.72±0.08 per cent in those of the latter ( P 0.02 Wilcoxon rank test). Conclusions: Despite similar environment conditions, placental apoptosis is increased in the smaller fetus and thus might play a role in discordancy between twins. Since increased placental apoptosis has also been found in singleton intrauterine growth restriction, this supports the hypothesis that the smaller twin is selectively growth restricted.

Nimodipine, a calcium-channel blocker that can cross the blood-brain barrier, has been shown to inhibit pulsatile GnRH release in vitro. We now show that oral nimodipine can effectively inhibit the ovarian cycle in mice in a dose-related manner.

Endometriosis: Science and Practice must be described as a definitive and all inclusive book on endometriosis. Expertly edited by Professors Linda Giudice, Johannes Evers, and David Healy, the book provides a complete overview of the knowledge gathered from the increased understanding of endometriosis over the past 20 years. Endometriosis is frequently intensely painful and can lead to infertility causing disruption of women’s lives and relationships. It can be a devastating condition. Major progress has been made in recent years on understanding the causes of endometriosis and developing effective clinical therapies. Endometriosis: Science and Practice fuses the scientific and clinical approaches. It presents the characteristics of the disease, how it presents, describes its possible causes and delineates the medical and surgical approaches to pain therapy, and wider aspects of the disease. Broad in outlook, focused in application, Endometriosis: Science and Practice covers pathogenesis, disease characterization, biological basis of endometriosis, diagnosis, medical therapies, surgical therapies, and its relation to infertility and associated disorders. While no cause for endometriosis has been fully determined, information of recent developments is outlined in this text, offering insight to improve management of symptoms medically or surgically. The first of its kind, this major textbook integrates scientific and clinical understanding of this painful disease helping to provide better patient care. This book is an excellent resource for physicians who want to keep up-to-date with recent advances in diagnosis and management of endometriosis as it provides a comprehensive approach to the biology, diagnosis, and treatment of endometriosis, pointing at the latest in molecular, genetic, and epigenetic research underlying the pathophysiology of endometriosis. It is a must read for every single person who is serious about moving the field of endometriosis forward. One-hundred-and-fourteen leading (and actively practicing) surgeons, physicians, researchers, as well as emerging leaders in the field of endometriosis have contributed with their insights into this complex disease providing very short links between the laboratory and clinical practice. This is the first time such a complete compilation has been published on endometriosis addressing not only basic research, diagnosis, and therapeutics, but also treatment outcomes and impact. The book concludes with an “eye to the future” in terms of emerging research, diagnostics, and therapeutics. Learners of all ages and from multiple disciplines, clinicians, researchers, and patients will benefit from the knowledge imparted in the pages of this book, which we hope will stimulate new knowledge, so one day we can cure endometriosis or, perhaps, even better, prevent it. The stellar cast of contributors, led by world-leading editors from the US, Europe, and Asia-Pacific, have produced a contemporary tour-de-force that should be on the bookshelf of anyone who provides medical care for women.

Objective Immature myeloid cells (IMCs) are bone marrow–derived cells that normally differentiate into granulocytes, macrophages, and dendritic cells (DCs) but expand in pathological conditions such as malignancy. DCs are antigen-presenting cells that regulate the immune response. Both IMCs and DCs were shown to take part in angiogenesis; however, little is known of their function in the placenta. We sought to determine whether alterations in DC and IMC populations in the placenta precede the onset of delivery. Study Design Experiments were performed on 6-8 week old C57Bl/6 female mice. Placentas from pregnant mice that were killed on designated days, immunostained using fluorescently labeled anti-CD11b, Gr-1, CD11c, major histocompatibility II (MHCII), and CD45, and analyzed by flow cytometry and immunofluorscent microscopy. Results Throughout the latter part of pregnancy toward labor and delivery, the CD45 + CD11b + Gr1 + -IMC population decreased 29 ± 9.1% (day 12) and 30 ± 9.9% (day 15), vs 21 ± 8.1% (day18, n = 21, 15, and 27; P = .006 and P = .004, respectively), whereas the CD45 + CD11c + MHCII + -DC population increased 0.87 ± 0.3% (day 12) and 0.70 ± 0.3% (day 15) vs 1.81 ± 1.3% (day 18, n = 21, 15, and 27, P = .002 and P = .001, respectively). Both myeloid cell populations were localized adjacent to CD31 + endothelial cells in sites of placental angiogenesis. Conclusion Labor and delivery are preceded by proangiogenic-myeloid cell alterations, reflected by a decrease in IMCs and an increase in DCs populating the mouse placenta. The intriguing possibility that delivery is preceded by the maturation of IMCs in part into DCs warrants further studies.