Distribution and function of 3',5'-Cyclic-AMP phosphodiesterases in the human ovary.

The concentration of the important second messenger cAMP is regulated by phosphodiesterases (PDEs) and hence an attractive drug target. However, limited human data are available about the PDEs in the ovary. The aim of the present study was to describe and characterise the PDEs in the human ovary. Results were obtained by analysis of mRNA microarray data from follicles and granulosa cells (GCs), combined RT-PCR and enzymatic activity analysis in GCs, immunohistochemical analysis of ovarian sections and by studying the effect of PDE inhibitors on progesterone production from cultured GCs. We found that PDE3, PDE4, PDE7 and PDE8 are the major families present while PDE11A was not detected. PDE8B was differentially expressed during folliculogenesis. In cultured GCs, inhibition of PDE7 and PDE8 increased basal progesterone secretion while PDE4 inhibition increased forskolin-stimulated progesterone secretion. In conclusion, we identified PDE3, PDE4, PDE7 and PDE8 as the major PDEs in the human ovary.

Impact of PCOS on early embryo cleavage kinetics.

This study investigated whether polycystic ovary syndrome (PCOS) affected early embryo development assessed by time-lapse analysis of embryo kinetics from fertilization to the blastocyst stage. This was a prospective cohort study of two pronuclei (2PN) embryos from 25 hyperandrogenic PCOS patients (110 2PN embryos), 26 normoandrogenic PCOS patients (140 2PN embryos) and 20 healthy, regularly cycling women (controls, 97 2PN embryos). Patients underwent the same baseline evaluation and the same ovarian stimulation from April 2010 to February 2013. Oocytes were fertilized by intracytoplasmic sperm injection and incubated in an EmbryoScope with pictures taken every 20 min in seven focal planes. Time to 2PN breakdown, first cleavage and cleavage to 3, 4, 5, 6, 7 and 8 cells, morula and blastocyst (t2, t3, t4, t5, t6, t7, t8, t(M), t(B)) were annotated. Differences in embryo kinetics between groups were assessed by mixed modelling. Compared with controls, embryos from hyperandrogenic PCOS patients were significantly delayed at 2PN breakdown, t2, t3, t4 and t7 but not at t5, t6, t8, t(M) or t(B). Embryos from hyperandrogenic PCOS women had developed slower from fertilization to the 8-cell stage compared with embryos from controls.

Identification of new ovulation-related genes in humans by comparing the transcriptome of granulosa cells before and after ovulation triggering in the same controlled ovarian stimulation cycle.

STUDY QUESTION: Which genes and molecular mechanisms are involved in the human ovulatory cascade and final oocyte maturation? SUMMARY ANSWER: Up-regulated genes in granulosa cells (GC) represented inflammation, angiogenesis, extracellular matrix, growth factors and genes previously associated with ovarian cancer, while down-regulated genes mainly represented cell cycle and proliferation. WHAT IS KNOWN ALREADY: Radical changes occur in the follicle during final follicle maturation after the ovulatory trigger: these range from ensuring an optimal milieu for the oocyte in meiotic arrest to the release of a mature oocyte and remodeling into a corpus luteum. A wide range of mediators of final follicle maturation has been identified in rodents, non-human primates and cows. STUDY DESIGN, SIZE, DURATION: Prospective cohort study including 24 women undergoing ovarian stimulation with the long gonadotrophin-releasing hormone agonist protocol during 2010-2012 at Holbæk Fertility Clinic. Nine paired samples of GC and 24 paired samples of follicular fluid (FF) were obtained before and after recombinant human chorionic gonadotrophin (rhCG) administration. PARTICIPANTS/MATERIALS, SETTING, METHODS: Nine paired (nine arrays before rhCG and nine arrays after rhCG) samples of GC mRNA were amplified and hybridized to Affymetrix Human Gene 1.0 ST GeneChip arrays, compared and bioinformatically analyzed. Eleven selected genes were validated by quantitative reverse transcriptase PCR. FF hormones were analyzed by enzyme-linked immunosorbent assay. MAIN RESULTS AND THE ROLE OF CHANCE: Eleven hundred and eighty-six genes were differentially expressed (>2-fold, P<0.0001, false discovery rate <0.0012) when comparing GC isolated before and 36 h after hCG, among those were genes known to be expressed at ovulation, i.e. ADAMTS1 and HAS2. Many new ovulation-related genes were revealed, such as CD24, ANKRD22, CLDN11 and FBXO32. FF estrogen, androstenedione and anti-Müllerian hormone decreased significantly while progesterone increased, accompanied by radical changes in the expression of steroidogenic genes (CYP17A, CYP19A, HSD11B1 and HSD11B2, StAR). Genes related to inflammation, angiogenesis, extracellular matrix formation, growth factors and cancer were up-regulated while cell cycle genes were massively down-regulated. Seventy-two genes previously described in connection with ovarian cancer were among the highly regulated genes. In silico analysis for top upstream regulators of the ovulatory trigger suggested--besides LH--TNF, IGF1, PGR, AR, EGR1 (early growth response 1), ERK1/2 (extracellular signal regulated kinase 1/2) and CDKN1A (cyclin-dependent kinase inhibitor 1A) as potential mediators of the LH/hCG response. LIMITATIONS, REASONS FOR CAUTION: The present dataset was generated from women under hormonal stimulation. However, comparison with a macaque natural cycle whole follicle ovulation dataset revealed major overlap, supporting the idea that the ovulation-related genes found in this study are relevant in the human natural cycle. WIDER IMPLICATIONS OF THE FINDINGS: These data will serve as a research resource for genes involved in human ovulation and final oocyte maturation. Ovulation-related genes might be good candidate biomarkers of follicle and oocyte health. Further, some of the ovulation-related genes may serve as future ovarian cancer biomarkers. STUDY FUNDING/COMPETING INTEREST(S): Grants from the Research Fund of Region Sjælland are gratefully acknowledged. None of the authors declared any conflict of interest. TRIAL REGISTRATION NUMBER: Not applicable.

Polycystic ovary syndrome and low-grade inflammation with special reference to YKL-40.

OBJECTIVE: To evaluate the plasma level of YKL-40 in a Danish polycystic ovary syndrome (PCOS) population and to investigate whether YKL-40 is associated with CVD risk factors such as waist circumference, body mass index (BMI), insulin resistance (IR), fasting glucose, fasting insulin, blood lipids and CRP. DESIGN: Cross-sectional study. SETTING: Gynecological clinics at three Danish University Hospitals. PATIENTS: One hundred seventy-one premenopausal women with PCOS recruited consecutively from April 2010 to February 2012. PCOS was diagnosed according to the Rotterdam criteria. MAIN OUTCOME MEASURES: Plasma level of YKL-40 in four phenotypes of PCOS defined by BMI and IR. RESULTS: No statistically significant difference was observed in the plasma level of YKL-40 across the four BMI/IR-phenotypes. Positive associations were observed between YKL-40 and BMI, total and free testosterone, triglycerides, and CRP. Total and free testosterone were independent predictors of YKL-40. CONCLUSION: YKL-40, the marker of low-grade inflammation is not increased in women with PCOS.

Polycystic ovary syndrome: infertility, cardiovascular, metabolic and obstetrical risks, laboratory and clinical outcomes -- the PICOLO Study.

OBJECTIVES: The primary objective of this multicenter study is to evaluate the relative impact of insulin resistance (IR) and body mass index (BMI) in women with polycystic ovary syndrome (PCOS) on (1) Key hemodynamic/thrombogenic variables, (2) Oocyte quality and early embryo development, (3) Fetal growth, placental function and adverse obstetric outcome. SECONDARY OBJECTIVE: To establish a PCOS database and biobank facilitating future basic and interventional research related to PCOS. DESIGN: A cross-sectional and longitudinal cohort study at four University Hospitals in Denmark. POPULATION INCLUSION: About 200 women fulfilling the Rotterdam Criteria and 100 women without PCOS recruited from 2010 to 2012. METHODS: The impact of PCOS, as well as the impact of IR and BMI on the hormonal, metabolic and hemostatic key variables will be analyzed combining conventional, molecular techniques and selected gene analysis. Oocytes will be characterized by gene expression of granulosa and cumulus cells and the early embryo development will be followed by time lapse microscopy. Fetal growth will be assessed by repeated ultrasound measurements, and the pregnancy outcome compared to maternal and fetal biochemical markers of growth and inflammation and clinical pregnancy complications. MAIN OUTCOME MEASURES: Metabolic and hemostatic risk-biomarkers, oocyte and embryo quality, adverse pregnancy outcome, fetal growth and placental function in women with PCOS.

[Experience about more than 2000 renal transplantations at the university of Brussels]. / A propos de 2000 greffes rénales a l'U.L.B.: le parcours d'un patient transplanté rénal en 2007.

Since 1965, more than 2000 renal transplantations (including more than 100 living-donor transplantations) have been performed at the University of Brussels. An end-stage renal disease patient candidate to renal transplantation will be therefore followed from his enrolment on the waiting list to the long-term post-transplant period. Improvement in the outcome of renal transplantation is achieved due to better knowledge in many fields of medicine, such as immunology, infectious disease, metabolic diseases (hyperlipemia, diabetes mellitus), pharmacology, use of immunosuppressive regimen, a more adequate cardiovascular prevention and treatment. If the best results were achieved with kidneys from living donors, the graft survival rate at the University of Brussels was nearly 80% for the last period (2000-2006). Unfortunately, renal transplantation cannot cure certain comorbid conditions and even may promote them: infectious diseases, neoplasia, metabolic disorders (e.a diabetes mellitus, hyperlipemia). Many efforts have to be done to develop less toxic and more immune selective therapeutic strategies. Living donation and extension of the pool of cadaveric donors will reduce the length of time spent on the waiting list and will significantly impact on mortality and morbidity after kidney transplantation.

Predictors of South African male testosterone levels: the THUSA study.

The process of urbanization occurring in many developing countries may have consequences for reproductive endocrine function. Here, we test predictions concerning variation in South African male testosterone levels among subgroups across an urbanization gradient representing differences in both geography and socioeconomic status. Subjects included 364 males aged between 20 and 82 comprising a cross-sectional study conducted between 1996 and 1998. Testosterone levels were measured from serum samples obtained between 08:00 and 11:00. In ANCOVA analysis, male testosterone levels differed significantly along this rural-to-urban gradient, with members of the most urban group having higher testosterone levels than groups of farmers and inhabitants of informal housing areas adjacent to towns. Testosterone levels declined with age and were negatively related to body mass index (BMI). Testosterone levels did not differ according to HIV status. Further exploratory ANCOVA analyses revealed that physical activity levels, depression, affect, and hostility were not significantly associated with variation in testosterone levels. These data help document causes of variation in male testosterone levels in a context of urbanization and may have implications for clinical outcomes such as the development of a male hormonal contraceptive or prostate cancer.

[The medico-surgical department of nephrology dialysis and renal transplantation]. / Le département médico-chirurgical de Néphrologie Dialyse et Transplantation Rénale.

The Department of Nephrology of the Hospital Erasme, opened 25 years ago, is now performing, each year, 22,000 hemodialysis sessions, 800 patient-weeks of peritoneal dialysis treatment and 70 renal grafts. Scientific contributions of the department deal with vascular access for hemodialysis, susceptibility to infections of dialyzed patients, parathyroid surgery, biocompatibility of dialysis membranes, predictive factors of renal graft survival, immunosuppression with monoclonal antibodies, experimental studies of graft tolerance and rejection, toxic nephropathies. The most original contributions are related to anaphylactoïd reactions in hemodialysis by association of acrylonitrile membranes with inhibition of the converting enzyme, to advantages and side effects of OKT3 monoclonal antibody and to discovery and study of the Chinese herbs nephropathy.

Fatal primary infection due to human herpesvirus 6 variant A in a renal transplant recipient.

We describe a fatal primary human herpesvirus 6 (HHV-6) variant A infection in a kidney transplanted adult woman. On day 20 post transplantation (TX), after rejection therapy, the patient presented an acute hemophagocytic syndrome with hepatitis and central nervous system involvement. HHV-6 IgG and IgM antibodies seroconversion was demonstrated. HHV-6 variant A was the sole pathogen detected by nested PCR and/or culture in blood, bone marrow aspiration, liver biopsy, cerebrospinal fluid and bronchoalveolar lavage. The graft was HHV-6 seropositive and the patient was not transfused before day 28 post TX, suggesting that the virus was transmitted by the graft. Despite immunoglobulins, ganciclovir and foscarnet therapy, the HHV-6 infection progressed and led to severe aplasia. The patient developed Aspergillus fumigatus pneumonia and died from fulminant candidemia. This case demonstrated for the first time that HHV-6 variant A primary infection can cause life-threatening disseminated infection in immunosuppressed patients.

An epidemiological study of hypertension and its determinants in a population in transition: the THUSA study.

BACKGROUND: Many black persons in South Africa have been subjected to urbanisation and urbanisation has led to a significant increase in diseases of lifestyle. The determinants of hypertension in a population in transition have not been well-defined and there is a pressing need for observational epidemiological studies as well as randomised-controlled trials in populations from Africa. The aim of this study was to investigate the association between blood pressure and factors known to contribute to hypertension. METHODS: The study sample consisted mainly of Setswana speaking people, divided into different levels (strata) of urbanisation, namely stratum 1 (rural) to stratum 5 (urbanized). A total of 1821 black subjects, which included 1040 woman, were recruited and randomly selected from 37 sites from the four geographical quarters of the North West Province of South Africa. The following questionnaires were used: demographic, anthropometric, quantitative food frequency, physical activity and scales to measure psychosocial variables. Biochemical analysis (standardised methods) were done on the serum and plasma of the subjects and the blood pressure was measured with a sphygmomanometer. RESULTS: Of the total sample, 22.8% of the subjects had systolic and 20.7% diastolic blood pressures above 140/90 mm Hg. Males and females from stratum 3 showed the highest rate of hypertension (32.9% systolic and 25.1% diastolic) and stratum 5 the lowest. Blood pressure correlated positively with age, level of urbanisation, WHR (waist:hip ratio) and smoking. In the woman the diastolic blood pressure correlated the best with body mass index (BMI), serum triglycerides, total serum cholesterol, low-density lipoprotein (LDL) cholesterol and s-GGT. Coping strategies, experience of social support, cultural aspects and affect balance are related to blood pressure, especially in the case of women. CONCLUSIONS: It seems that factors associated with urbanisation are related to the manifestation of hypertension in black people of the North West Province, given the highest mean blood pressure in people living in informal settlements, where most newcomers to the urban areas live. Journal of Human Hypertension (2000) 14, 779-787

[Dioxins: current knowledge about health effects]. / Les dioxines: état actuel des connaissances des effets sur la santé.

Dioxins are a family of chlorinated aromatic hydrocarbons that are produced during combustion processes in the presence of a chlorine donor and as by-products of the chlorine-processing chemical industries. Several dioxins are extremely stable compounds and persist for years in the environment. Exposure to dioxins occurs mainly via the ingestion of contaminated food. The lipophilic character of dioxins prevents their excretion in the urine and causes their accumulation in body fat. The mechanisms of dioxin action are similar to those of a hormone. Dioxins bind to a specific intracellular receptor and the complex acts as a transcription factor that induces the production of a great number of proteins. Certain dioxins, particularly 2,3,7,8-tetrachlorodibenzo-p-dioxin, are very toxic and able to induce numerous clinical conditions. The carcinogenicity of dioxins is well documented in animal models and has been described in humans after professional and accidental exposures. Recent experimental data also indicate that dioxins can cause dysfunction of the sexual and thyroid hormone systems and that the administration of dioxins induces several conditions related to hormonal dysfunction. Chronic exposure of female Rhesus monkeys increases the incidence and severity of endometriosis. The administration of dioxins during pregnancy and nursing causes altered development of the reproductive system, decreased spermatogenesis, hypothyroidism and disturbed psychomotor development in the offspring. The particular sensibility of the fetus and newborn is of concern because the exposition to dioxins is particularly important during those periods of life. In humans a series of conditions related to hormonal dysfunction as undescended testis, decreased spermatogenesis, testicular cancer and endometriosis have increased in incidence during the last decades. The chronological parallelism with the appearance of dioxins in the environment suggests that these might exert biological effects at the prevailing level of exposure. Nevertheless this hypothesis is currently unconfirmed by epidemiological studies. The implications of this scientific incertitude for the implementation of preventive measures are briefly discussed.

A role for Th2 cytokines in the suppression of CD8+ T cell-mediated graft rejection.

A major histocompatibility complex (MHC) class I-specific T cell receptor (TCR)-transgenic mouse was used to study classical-type transplantation tolerance in the adult. Engraftment of MHC class I-incompatible bone marrow and tolerance to donor-type skin grafts were obtained using dimethylmyeleran (DMM) as a myeloablative agent and a non-depleting anti-CD8 monoclonal antibody (mAb) as the sole immunosuppressant. Surprisingly, bone marrow engraftment was facilitated by host CD4+ T cells, a subset normally considered unable to reject class I MHC-incompatible grafts. A combination of mAb to interleukins (IL)-4 and -10 antagonized the "permissive" effects of host CD4+ T cells, indicating a possible role for Th2-type immunoregulation that can act on CD8+ T cells in this form of transplantation tolerance. The fate of graft-reactive T cells was monitored using anti-clonotypic antibodies. It was observed that bone marrow engraftment then led to peripheral deletion of mAb-blockaded, clonotype+ CD8+ T cells.

Hepatic MR imaging: comparison of RARE derived sequences with conventional sequences for detection and characterization of focal liver lesions.

BACKGROUND: We compared two T2-weighted turbo spin echo (TSE) sequences with a T2-weighted conventional SE (CSE) sequence to determine whether sequences derived from rapid acquisition with relaxation enhancement such as TSE could replace CSE for the detection and subsequent characterization of focal liver lesions. METHODS: A total of 55 consecutive patients with 107 liver lesions underwent magnetic resonance imaging examinations at 1.5 Tesla, with a constant imaging protocol. TSE pulse sequences were acquired with eight echo trains (repetition time [TR], 4718 ms; echo time [TE], 90 ms; acquisition time [TA], 4.03 min; and a symmetric k-space ordering scheme) and 11 echo trains (TR, 4200 ms; TE, 140 ms; TA, 4.40 min; and an asymmetric k-space ordering scheme) and compared with CSE (TR, 2300 ms; TE, 45/90 ms; TA, 9.53 min). Images were analyzed qualitatively by scoring image quality and artifacts and counting focal liver lesions by independent reading with consensus obtained for discrepancies. Quantitative analysis was performed by measuring signal-to-noise (S/N), contrast-to-noise (C/N), and tumor-liver signal intensity (T/L) ratios. RESULTS: T2-weighted TSE sequences provided better subjective image quality and reduced artifacts as compared with the T2-weighted CSE sequence. CSE and TSE sequences exhibited no statistically significant differences in liver S/N, lesion-liver C/N (CSE TE, 90 ms: 18.6 +/- 14.0; TSE TE, 90 ms: 16.5 +/- 12.9) and the detectability of focal liver lesions. Heavily T2-weighted TSE with a TE of 140 ms allowed correct characterization of focal liver lesions based on a T/L ratio of 3.0 in 84% of patients. CONCLUSIONS: T2-weighted TSE sequences are as suited as CSE for the detection (TE, 90 ms), and appear to be superior for the characterization (TE, 140 ms), of focal hepatic lesions. Whether a single sequence, such as a double-echo TSE or a single-echo TSE sequence with a TE between 110 and 120 ms, might perform both functions as well or better than CSE is unknown. However, because of time savings, TSE eventually may be preferred over CSE.

Modulation of the release of cytokines and reduction of the shock syndrome induced by anti-CD3 monoclonal antibody in mice by interleukin-10.

Since IL-10 was recently shown to inhibit several T cell functions in vitro, we investigated the effects of IL-10 on the cytokine release syndrome induced in mice by the 145-2C11 anti-CD3 mAb. As OKT3 in man, this mAb induces a massive polyclonal T cell activation before to induce immunosuppression. First, we found that administration of 1000 U of recombinant mouse IL-10 (mIL-10) 30 min before injection of 10 micrograms of the 145-2C11 antimouse CD3 mAb markedly reduced the systemic release of IFN-gamma and TNF. In contrast, IL-10 pretreatment did not significantly modify the release of IL-6. To determine the effect of IL-10 pretreatment on the endogenous secretion of IL-10 induced by the 145-2C11 mAb, mice were injected with human IL-10 (hIL-10) which does not cross-react in the ELISA for mIL-10 determination. While hIL-10 was as efficient as mIL-10 in reducing TNF and IFN-gamma release, it did not modify peak serum levels of IL-10. The modulation of cytokine production by mIL-10 was associated with a significant reduction of the toxicity of the 145-2C11 mAb, as assessed by the attenuation of hypothermia and by the reduced lethality in D-galactosamine-sensitize mice. We conclude that IL-10 differentially regulates the in vivo production of cytokines and decreases the systemic toxicity induced by the 145-2C11 mAb. These observations suggest potential therapeutic applications of IL-10 in organ transplantation, especially in association with anti-CD3 mAb.