Preovulatory Phase Research Articles

Myeloid-related protein-8/14 is associated with proinflammatory cytokines in cervical mucus

Myeloid-related protein-8 (MRP-8), MRP-14, and MRP-8/14 are found in a variety of inflammatory conditions and are involved in the host defense system. The objective of this study was to determine the concentrations of MRP-8, MRP-14, and MRP-8/14 in human cervical mucus and the associations between MRP-8/14 and proinflammatory cytokines. Samples of cervical mucus were obtained using a syringe from sexually active women ( n = 97) during the preovulatory phase. Samples from seven women were obtained using a swab placed in the cervical canal during the proliferative, preovulatory, and luteal phases. Concentrations of MRP-8, MRP-14, MRP-8/14, IL-1α, IL-6, IL-8, and granulocyte elastase were measured using an ELISA. The mean levels of MRP-8, MRP-14, and MRP-8/14 in cervical mucus were 1.87, 0.46, and 23.90 μg/ml, respectively. The concentration of MRP-8/14 showed positive correlations with concentrations of IL-1α ( p < 0.0001), IL-8 ( p < 0.0001), and granulocyte elastase ( p < 0.0001). However, there were no significant differences in MRP-8/14 levels in the cervical mucus of each patient during the menstrual cycle. MRP-8/14 was mainly detected in human cervical mucus and showed a positive correlation with proinflammatory cytokines. The MRP-8/14 level in cervical mucus may be useful as a marker of inflammation of the uterine cervix.

The Relationship between Menstrual Cycle Phase and Anterior Cruciate Ligament Injury

Background Female athletes suffer a greater incidence of anterior cruciate ligament tears compared with male athletes when participating in common sports; however, very little is known about the factors that explain this disparity. Study Design Case-control study; Level of evidence, 3. Methods Female recreational alpine skiers with an anterior cruciate ligament rupture and age-matched control skiers provided a serum sample and self-reported menstrual history data immediately after injury. Both serum concentrations of progesterone and menstrual history were then used to group subjects into either preovulatory or postovulatory phases of the menstrual cycle. Results Analysis of serum concentrations of progesterone revealed that alpine skiers in the preovulatory phase of the menstrual cycle were significantly more likely to tear their anterior cruciate ligaments than were skiers in the postovulatory phase (odds ratio, 3.22; 95% confidence interval, 1.09-9.52; P = .027). Analysis of menstrual history data found similar results, but the difference was not statistically significant (odds ratio, 2.38; 95% confidence interval, 0.86-6.54; P = .086). Conclusion The likelihood of sustaining an anterior cruciate ligament injury does not remain constant during the menstrual cycle; instead, the risk of suffering an anterior cruciate ligament disruption is significantly greater during the preovulatory phase of the menstrual cycle compared with the postovulatory phase. Clinical Relevance Phase of menstrual cycle may be one of the risk factors that influence knee ligament injury among female alpine skiers. The findings from this study should be considered in subsequent studies designed to identify persons at risk for anterior cruciate ligament injury and to develop intervention strategies.

Up-regulated mRNA expression of Toll-like receptor (TLR)-2 and -4 in bovine oviductal cells after treatment with estradiol or progesterone indicates a stimulatory effect of steroid hormones on the immune system in the reproductive tract

Female reproductive epithelial cells produce a variety of substances that promote reproduction and immune defense. These cells secrete peptide mediators that regulate the traffic and activity of immune cells. Several factors are supposed to be involved in these processes, e.g. Toll-like receptors (TLR) which mediate the induction and secretion of chemokines after recognizing pathogens. The aim of the present study was to investigate the mRNA expression of TLR-2 and TLR-4 in cultured bovine oviductal cells after treatment with steroid hormones as well as in different regions of the bovine oviduct during the estrous cycle in vivo. Bovine oviducts from the slaughterhouse were classified into four phases: pre-ovulatory, post-ovulatory, early or late luteal phase. Oviductal epithelial cells (>60%) were harvested by flushing separately the ampulla and the isthmus. Furthermore, primary cell culture was carried out with attached oviductal cells after 4 days. Then, the cells were treated with 10 pg/ml estradiol or 10 ng/ml progesterone for up to 6h, respectively. Total RNA was extracted from oviductal cells and real-time RT-PCR was performed. Treatment of cultured oviductal cells with estradiol or progesterone resulted in a significant 5–8 fold higher expression of TLR-2 and TLR-4 after 4h or 6h. Analysis of oviductal cells in vivo revealed a significant lower mRNA expression of TLR-2 during the pre-ovulatory phase compared with the post-ovulatory phase. Interestingly, a higher local TLR-2 expression was noted after ovulation in the isthmus than in the ampulla. In contrast, TLR-4 expression seems to be unregulated without regional differences during the whole estrous cycle in vivo. These results showed that TLR-2 and TLR-4 were expressed in the bovine oviduct. Especially, the regulated expression of TLR-2 and the stimulation by steroid hormones suggests that this receptor is important for an oviductal immune response. Such results support the hypothesis that steroid hormones have an impact on the immune system in reproductive tissues.

Effects of a preovulatory single low dose of mifepristone on ovarian function

Objectives To investigate the effect of a single low dose of mifepristone on ovarian function, when administered in the preovulatory period.Methods Healthy women with regular menstrual cycles were studied during two consecutive menstrual cycles. Either mifepristone or placebo was given in a randomized double-blind order when the leading follicle reached a diameter between 15 and 17 mm. Daily ultrasound and serum hormone measurements were obtained until follicular collapse. Statistical analysis was performed using Wilcoxon signed-rank test.Results Eight women entered the study, although one woman had to be excluded afterwards from analysis because her LH surge had already appeared on the day of treatment. The LH surge was delayed from day 14 to 17 (P = 0.01). Mifepristone caused a 3-day delay in follicular collapse, occurring on day 16 in control cycles and on day 19 in mifepristone treatment cycles (P = 0.02). The median cycle length was 26 days in control cycles and 30 days in mifepristone treatment cycles (P = 0.03). Progesterone measurement 7 days after follicular collapse did not differ significantly between both cycles.Conclusions A single 10-mg dose of mifepristone administered during the preovulatory phase of the cycle delays the LH surge and postpones ovulation.

Pattern of prolactin secretion after administration of gonadotropin-releasing hormone agonist at the preovulatory phase of intrauterine insemination cycles

Administration of a gonadotropin-releasing hormone (GnRH) agonist at the preovulatory phase is an option for triggering ovulation in assisted reproductive technology cycles. The aim of this work was to investigate the pattern of prolactin secretion after the administration of a single dose of GnRH-agonist at the preovulatory phase. Descriptive study at a tertiary referral center. Fifteen normally ovulating patients undergoing ovarian stimulation for intrauterine insemination were studied. Ovarian stimulation was carried out using human menopausal gonadotropin (intramuscular 75 IU daily). When at least one follicle reached 17 mm (observed echographically), 0.5 mg of buserelin acetate was administered. Blood samples were taken to determine prolactin concentrations, at the time of agonist injection and 4, 8, 12, 24 and 48 hours later. A statistically significant increase in serum levels of prolactin was observed 4, 8 and 12 hours after GnRH-agonist administration, with a peak at 8 hours. The administration of a single dose of GnRH-agonist at the preovulatory phase in patients undergoing ovarian stimulation performed with human menopausal gonadotropin causes a significant increase in serum prolactin levels.

Development of the uterine shell glands during the preovulatory and early gestation periods in oviparous and viviparous Lacerta vivipara

The evolutionary process leading to the emergence of viviparity in Squamata consists of lengthening the period of egg retention in utero coupled with marked reduction in the thickness of the eggshell. We used light microscopy and scanning electron microscopy to study uterine structure during the reproductive cycle of oviparous and viviparous females of the reproductively bimodal Lacerta vivipara. We compared the structure of the uterine shell glands, which secrete components of the eggshell, during preovulatory and early gestation phases of the reproductive cycle and also compared histochemistry of the eggshells. The uterine glands of both reproductive forms undergo considerable growth within a period of a few weeks during folliculogenesis and vitellogenesis preceding ovulation. The majority of the proteinaceous fibers of the shell membrane are secreted early in embryonic development and the uterine glands regress shortly thereafter. This supports previous observations indicating that, in Squamata, secretion of the shell membrane occurs very rapidly after ovulation. The most striking differences between reproductive modes were larger uterine glands at late vitellogenesis in oviparous females, 101 microm compared to 60 microm in viviparous females, and greater thickness of the shell membrane during early gestation in oviparous females (52-73 microm) compared to viviparous females (4-8 microm). Our intraspecific comparison supports the conclusions of previous studies that, prior to ovulation, the uterine glandular layer is less developed in viviparous than in oviparous species, and that this is the main factor accounting for differences in the thickness of the shell membrane of the two reproductive forms of squamates.

Catamenial Epilepsy: The Elusive Condition

Frequency of Catamenial Seizure Exacerbation in Women with Localization-related Epilepsy Herzog AG, Harden CL, Liporace J, Pennell P, Schomer DL, Sperling M, Fowler K, Nikolov B, Shuman S, Newman M Ann Neurol 2004;56:431–434 This investigation assessed the frequency of catamenial epilepsy in 87 women who charted seizures and menses during three cycles. Catamenial epilepsy designation was made if two of three cycles showed at least one of three previously defined catamenial patterns. Among ovulatory cycles, average daily seizure frequency was significantly greater during the perimenstrual and preovulatory phases. Among anovulatory cycles, average daily seizure frequency was substantially less during the midfollicular phase than during the remainder of the cycle. Overall, 39.1% of the women had catamenial epilepsy.

Sex cycle modulates cancer growth

Among premenopausal women, both post-resection metastatic potential and tumor growth rate are influenced by the menstrual cycle. There is strong support for the former in large retrospective studies of surgical resection timing within the menstrual cycle and the following experiments were conducted to critically evaluate the latter. We studied a transplantable breast cancer of C3HeB/FeJ mice (3 studies), and a transplantable methylcholantherene A induced sarcoma of CD2F1 mice (2 studies). We concurrently measured local cancer size and estrous cycle stage up to twice and at least once each day. There is a natural individual variability in the average length of normal estrus (3-1/2 to 7 days) cycle in mice. We assessed the effect of the cycle stage and cycle duration on tumor size. We found identical estrous cycle stage coordination of cancer size, and identical effects of cycling frequency across all studies in each of these two tumors, both of which express both estrogen receptor alpha and progesterone receptor. Little or no change in cancer size occurs during proestrus (preovulatory phase) and estrus (periovulatory phase); tumor size increases several fold during diestrus (post-ovulatory phase); and the tumor shrinks partially as the next proestrus phase is approached. Across both mouse strains and tumor types, mice whose average cycle length is briefer (faster cyclers), have slower average tumor growth rate than those with longer cycles (slower cyclers) who have faster tumor growth rates. The virtually identical modulation of tumor size and cancer growth rate, in each of two very different transplantable cancers (one, classically sex-hormone-dependent, and the other, never previously recognized as hormone dependent) growing in two unrelated inbred mouse strains, indicates that the fertility cycle related host factors affect cancer size and growth rate. These experimental findings suggest that cancer cell proliferation of both breast and non-breast cancers in premenopausal women may be meaningfully coordinated by the menstrual cycle. If this proves to be the case, then any therapeutic strategy targeting proliferating cancer cells should be most effective against cancer of cycling women when given during the follicular phase of their menstrual cycles.

The Fallopian tubes in domestic mammals: how vital is their physiological activity?

Set in an historical perspective, this essay examines diverse physiological aspects of Fallopian tube function in domestic animals and man. Microsurgical experiments are described that established the role of the isthmus in imposing a sperm gradient up to the site of fertilisation. Resection of the isthmus followed by reanastomosis of the remaining portions of the tube generated a high incidence of polyspermy in mated animals. Scanning electron microscopy and surgical studies revealed that spermatozoa were arrested and stored in the caudal portion of the isthmus before ovulation, the so-called functional sperm reservoir. There were specific adhesion contacts between the sperm head and endosalpingeal microvilli or cilia. Further experiments indicated that very large numbers of competent spermatozoa could be released from pre-ovulatory binding by microinjections of a solution of progesterone in oil under the serosal layer of the tube: when suitably timed, such treatment led to a high incidence of polyspermic fertilisation. Avid sperm binding in the caudal isthmus before ovulation prevents myosalpingeal activity leading to abnormal fertilisation, as might occur with multiple mating. Temperatures in the reproductive system were assessed and the caudal isthmus was found to be cooler than the ampulla during the pre-ovulatory phase of sperm storage. Finally, the existence of fluid microenvironments within the Fallopian tubes was reported, and the role of suspended cumulus-corona cells in amplifying signals from the zygote examined. An impact of Fallopian tube fluids on embryonic gene expression was also considered--an influence that may be further imposed if such fluids have access to the uterine lumen.

Cobalt complex with GnRH stimulates the LH release and PKA signaling pathway in pig anterior pituitary cells in vitro

Metal complexes with GnRH were shown to interact with GnRH receptors in pituitary cells. In the present study we examined the effects of GnRH and its cobalt complex form (Co-GnRH) on LH secretion and generation of second messengers, namely inositol phosphates (IPs) and cAMP, in porcine pituitary cells in vitro. The cells were obtained from gilt pituitary at the pre-ovulatory phase of estrous cycle and cultured for 72 h before challenge with GnRH or Co-GnRH. Both substances induced a significant increase in LH release that was detectable after 60 min ( P < 0.05) of treatment, with the Co-GnRH complex being more efficient than GnRH at 180 min ( P < 0.01). GnRH and Co-GnRH were equally effective at 10 −8 M ( P < 0.01), however, at the lowest (10 −9 M) as well as the highest (10 −7 M) concentrations tested, Co-GnRH was more potent than its native counterpart ( P < 0.01). Interestingly, Co-GnRH revealed twice more efficient than GnRH at stimulating cAMP production, an effect which was detectable in cells after 1 h-incubation ( P < 0.001). In contrast, while native GnRH induced a rapid increase ( P < 0.05) in IPs no such effect of Co-GnRH was observed. These data demonstrate that Co-GnRH and GnRH differentially effect on the signaling pathway in porcine gonadotropes and suggest that in these cells, the releasing action of Co-GnRH results from the mediation via the cAMP/protein kinase A second messenger system.

Steroid receptor expression in late follicular phase endometrium in GnRH antagonist IVF cycles is already altered, indicating initiation of early luteal phase transformation in the absence of secretory changes

Ovarian stimulation for IVF profoundly alters the early luteal phase endometrial development. It has been hypothesized that this process has already started in the late follicular phase, as the endometrium has already been exposed to high steroid concentrations since that phase. The aim of the present study was to prospectively investigate the effect of multi-follicular ovarian stimulation for IVF on the late follicular phase endometrium histology and the expression of estrogen receptor (ER) and progesterone receptor (PR). In a cross-over study, 11 infertile women with normal ovulatory function, participating in an IVF programme and treated with GnRH antagonist/recombinant FSH ovarian stimulation, were enrolled in the study. Endometrial biopsies were taken in a natural cycle on the day of the onset of the surge of the LH, and in a subsequent stimulation cycle on the day of hCG administration for final oocyte maturation. Endometrial histological dating was carried out according to Noyes' criteria. Immunohistochemistry was performed, using commercially available antibodies for ER and PR endometrial expression. The immunohistochemical signal was recorded in 1000 epithelial cells in each compartment (glands and stroma). Endometrial expression for each of the two receptors was graded on a scale of 0-3, based on the intensity of nuclear staining. Then a score range between 0 and 3000 was recorded, and expressed as a mean score per 1000 stroma or glandular cells per sample (range: 0-3). Histological examination of biopsies both in natural and stimulated cycles showed no secretory changes. However, in stimulated cycles, PR expression was significantly up-regulated compared to natural cycles in both glands (1.67 versus 1.34, P < 0.05) and stroma (1.98 versus 1.62, P < 0.05), whereas ER was down-regulated in glands (1.15 versus 1.43, P < 0.05). In IVF cycles, the progesterone measurements, although within normal values (range 0.8-1.4 microg/l), were significantly higher than in natural cycles (0.99 vs 0.63 microg/l, respectively, P = 0.008). An ongoing pregnancy rate of 37.5% was achieved in the stimulated cycles. Although the current study found no early secretory transformation in stimulated endometria before hCG administration, the ER and PR expression in these endometria is similar to the one described during the first days of the luteal phase in natural cycles. Supraphysiological concentrations of estradiol and subtle progesterone rises in the late follicular phase might be responsible for this modulated steroid receptor profile. This phenomenon indicates accentuated maturation of the endometrium in IVF cycles from the pre-ovulatory phase onwards.

Cellular localization and changes in expression of prolactin receptor isoforms in sheep ovary throughout the estrous cycle

The actions of prolactin (PRL) on target cells depend on the type of prolactin receptor (PRLr) predominantly expressed, particularly whether the long PRLr isoform is expressed. The aims of this study were to determine the cellular localization and the changes in expression of long and short PRLr isoforms in sheep ovary throughout the estrous cycle. Long and short PRLrs were localized mostly in the same ovarian cells. Maximum signal intensity, particularly for long PRLrs, was found in stromal cells surrounding primordial and primary follicles, and, for both PRLrs, in granulosa cells of preantral follicles and in luteal cells. Moderate signal intensity for PRLrs was found in theca cells of preantral to ovulatory follicles, and in granulosa cells of antral follicles up to the gonadotropin-dependent stage. Decreasing immunoreactivity to PRLrs was found in granulosa cells of gonadotropin-dependent to ovulatory follicles. For long PRLrs in particular, no signal was found in mural granulosa cells of gonadotropin-dependent follicles; for both isoforms, no signal was found in most granulosa cells of ovulatory follicles. In primordial to gonadotropin-dependent follicles, cellular localization of PRLr was similar on days 0, 10 and 15 of the cycle. Oocytes consistently showed positive immunostaining for PRLrs. Comparative RT-PCR analysis of long and short PRLr expression showed that the short isoform is evenly expressed throughout the estrous cycle, whereas the expression of the long form increases at the time of estrus and decreases at mid-luteal phase and at the onset of the follicular phase. Expression of long PRLrs was greater than that of short PRLrs on day 0 of cycle; expression of both isoforms was similar on day 10 and on day 15, long PRLrs expression was lower than that of short PRLrs. Our results indicate that in sheep ovary, the maximum responsiveness to PRL might occur during the preovulatory phase of the estrous cycle.

Late follicular phase endometrium histology and steroid receptor expression in stimulated cycles for in vitro fertilization/embryotransfer treatment (IVF/ET) with a gonadotropin releasing hormone antagonist/recombinant follicle stimulating hormone (GnRH antagonist/recFSH) protocol

To study the effect of multifollicular ovarian stimulation for IVF/ET on the histology and the steroid receptors expression of estrogen (ER), progesterone (PR) and androgen (AR) in preovulatory endometrium. Prospective cross-over study. Ten infertile women with normal ovulatory function participating in IVF/ET program were enrolled in this study. Endometrial biopsies were taken in a natural cycle on the day of the onset of the surge of the luteinizing hormone (LH), and in a subsequent stimulation cycle on the day of human chorionic gonadotropin (HCG) injection for final oocyte maturation (both of these dates are designated as day LH0). Immunohistochemistry was performed using commercially available antibodies for evaluating ER, PR and AR endometrial expression. Metric variables were tested with the t-test for independent samples and t-paired test for non-independent variables while Fisher’s exact test was used to analyse nominal variables. All tests were two-tailed with a confidence level of 95% (p<0.05). As it was expected, in the stimulation cycles estradiol (1826 vs. 254 ng/L) and progesterone (1.0 vs. 0.7 μg/L) levels were statistically significant higher compared to natural cycles. Histological examination of biopsies both in natural and stimulated cycles showed no early secretory phenomena. However, in stimulated cycles PR expression was up-regulated in both glands (1.61 vs. 1.29, p<0.05) and stroma (1.95 vs. 1.53, p<0.05) whereas ER were down-regulated in both compartments of the endometrium (1.37 vs. 1.20 in glands, and 1.20 vs. 1.01 in stroma, P>0.05 respectively). AR immunostaining was almost absent in glands in both natural and stimulated cycles and there was no difference in the expression in stroma cells during stimulation. An ongoing pregnancy rate of 42.8% was observed (in three women no embryo was transferred either because of poor embryo quality or ovarian hyperstimulation syndrome). In endometrial receptivity investigations, cross-over studies are valuable as thus the interpatient variability is eliminated. Moreover by taking the day of LH surge as a reference point for your biopsy timing, dating accuracy can be improved. Although in our study we found no early secretory changes in stimulated endometria just prior the HCG injection, the ER and PR phenotype of these endometria, however, resembles the phenotype observed the first days of the luteal phase. This phenomenon indicates accentuated maturation of the endometrium in IVF stimulated cycles even since the preovulatory phase. The possible pathogenetic mechanism for this abnormality is the supraphysiological concentrations of estrogens and the subtle progesterone rise, before the ovulation triggering in stimulated cycles. Taking into account the evidence that extreme advanced endometrium maturation on the day of the oocyte retrieval has been associated with adverse pregnancy outcome, seems that endometrium records interventions during the follicular phase as well, which might have implications during the implantation window later in the luteal phase. In order to understand the critical period for human implantation especially during the stimulation cycles for infertility treatment, attention should be paid equally in both luteal and follicular phase parameters.

The effect of clomiphene citrate on uterine and ovarian blood flow in patients with unexplained infertility. Does estrogen and progesterone treatment reverse these changes?

It is well-known that clomiphene citrate (CC) treatment has possible adverse effects on endometrial development. However it is not known whether changes in the uterine or ovarian blood flow or changes in hormonal values are related to this phenomenon. The purpose of this study was to evaluate whether CC treatment causes uterine and ovarian blood flow changes and if so whether the use of preovulatory estrogen and luteal progesterone treatment restores them. Prospective longitudinal study in patients with unexplained infertility who were planned to undergo CC treatment. A total of 51 patients were included in this study. In their natural cycles women underwent preovulatory and mid-luteal uterine and ovarian artery blood flow (Pulsatility Index=PI, Resistance Index=RI) measurements, preovulatory estradiol, mid luteal progesterone assays and preovulatory and mid luteal endometrial thickness measurements. Then these values were compared with CC (100mg/day × 5 day) treatment cycles. Patients who were found to have one or more deteriorated parameters after CC treatment (endometrium <6 mm; luteal protesterone <10 ng/ml; PI 3; RI 0.9 ) were treated with conjugated estrogen(CEE) 0.625 mg/daily (9–14th day) and micronised vaginal progesterone(MP) 200mg/day in the luteal phase of the subsequent CC cycle. Mean values of preovulatory estradiol was significantly higher and preovulatory PI was significantly lower with CC treatment (p<0.05). Mean values of other parameters were not different in natural and CC cycles. There were no patients with abnormal values in their natural cycles, however CC treatment resulted in abnormal parameters in 23/51 (46%) of patients. Treatment with CC+CEE+MP resulted in restoration of abnormal parameters in 16/23 (65.2%) of patients. CC treatment causes uterine and ovarian blood flow alterations and has a deleterious effect on luteal hormonal values in almost half of the patients. This effect can be reversed by estrogen and progesterone treatments. Doppler evaluation in the preovulatory and luteal phase is a useful adjunct to detect the patients who can benefit of such a treatment.

Efeito da inserção de implante anticoncepcional contendo acetato de nomegestrol sobre a função ovariana, muco cervical e penetração espermática

OBJETIVOS: estudar o efeito de um unico implante de acetato de nomegestrol (Uniplant) sobre funcao ovariana, producao do muco cervical e penetracao espermatica, quando inserido na fase pre-ovulatoria. METODOS: estudo clinico aberto, comparativo, incluindo 20 mulheres com ciclos menstruais regulares que foram estudadas durante um ciclo menstrual antes (controle) e um ciclo menstrual depois da insercao do implante. Dosagens de hormonio luteinizante (LH), estradiol e progesterona, ultra-sonografia vaginal, coleta de muco cervical e teste de penetracao espermatica foram realizados. Para comparacao estatistica, foi utilizado o Student t-test para grupos pareados e o teste Wilcoxon nao parametrico. Os valores estao mostrados como medias ± erro padrao. RESULTADOS: todos os ciclos controles foram ovulatorios com parâmetros normais. Os niveis pre-ovulatorios de estradiol e LH diminuiram significativamente de 603,2 ± 78,0 pmol/l e 22,5 ± 6,5 UI/l na pre-insercao do implante para 380,7 ± 51,9 pmol/l e 4,9 ± 1,3 UI/l 48 horas apos a insercao (p < 0,05 e p < 0,01, respectivamente). Os niveis de progesterona mantiveram-se sem variacao significativa apos a insercao do implante (ciclo controle = 49,8 ± 3,3 nmol/l e ciclo tratado = 43,2 ± 5,2 nmol/l). Muco cervical e penetracao espermatica foram profundamente afetados em 10,5% das usuarias apos 20 horas da insercao do implante, em 68,5% das usuarias apos 24 horas e em 100% apos 48 horas. Ruptura folicular ocorreu na maioria dos ciclos tratados apos 48 horas da insercao. CONCLUSOES: o uso de um unico implante de acetato de nomegestrol inserido na fase pre-ovulatoria afetou os picos pre-ovulatorios de estradiol e LH, a producao do muco cervical e a penetracao espermatica, mas nao foi capaz de impedir a ovulacao, o que reforca a necessidade da insercao do implante nos 5 primeiros dias do ciclo menstrual.

Increased expression of cyclooxygenase-1 (COX-1) and -2 mRNA after ovulation in the bovine oviduct

Optimal conditions for fertilization and development of the early embryo is provided by the mammalian oviduct. Prostaglandins (PGs) are present in the bovine oviduct playing an important role in muscle contraction or as survival factors. COX-1 and -2 are key enzymes in the synthesis of prostaglandins by converting arachidonic acid into PGH2 as a precursor of various PGs. The aim of the present study was to investigate the mRNA expression of COX-1 and COX-2 in different regions (ampulla and isthmus) of the bovine oviduct during the estrous cycle. Bovine oviducts were collected at the slaughterhouse throughout the estrous cycle and classified into the following four phases: pre-ovulatory, post-ovulatory, early or late luteal phase. The obtained oviducts were divided in ipsilateral (to ovulation/dominant follicle) or contralateral oviducts. Oviductal epithelial cells were harvested by flushing separately the ampulla and the isthmus. Total RNA was extracted and semi-quantitative RT-PCR was performed followed by quantitative real-time RT-PCR. Expression of COX-1 and -2 mRNA was detected in all regions of the oviduct during the whole estrous cycle. Quantitative real-time RT-PCR revealed a significant higher expression of COX-2 in the ampulla compared with the isthmus during all stages of the estrous cycle. The highest expression of COX-2 was observed in the ipsilateral ampulla directly after ovulation. In contrast, COX-1 showed a twofold higher mRNA expression after ovulation compared with the other estrous cycle phases. No difference between ipsilateral and contralateral oviducts nor between the ampulla and the isthmus was noted. These results showed that both COX were differently expressed in a cycle stage-dependant manner. Especially the expression pattern of COX-2 suggests that an estrous cycle-dependant and region-specific synthesis of PGs is required for a successful reproduction. Therefore, we hypothesize that a local PG synthesis may directly influence survival and transport of the ovulated egg as well as of the early embryo.

Serum pattern of circulating free leptin, bound leptin, and soluble leptin receptor in the physiological menstrual cycle

Objective To investigate the serum pattern of free leptin, bound leptin, and soluble leptin receptor throughout the physiological menstrual cycle. Design Prospective observational study. Setting Tertiary care center for gynecological endocrinology and reproductive medicine and a university research laboratory. Patient(s) Thirty regularly cycling volunteers (age, 29 ± 4.2 years). Intervention(s) Blood sampling was performed at different phases (early and mid follicular phase, preovulatory phase, and early and late luteal phase) of three consecutive menstrual cycles; each phase of the menstrual cycle was investigated twice. Main outcome measure(s) Free leptin, bound leptin, soluble leptin receptor, LH, E 2, P, vaginal ultrasound. Result(s) A peak of serum free leptin levels was found in the late luteal phase followed by a significant drop in the early follicular phase and again by a continuous increase up to the next luteal peak. There were no significant alterations in serum bound leptin and soluble leptin receptor levels. Conclusion(s) The present study shows that there are significant circacyclic fluctuations of free leptin levels with the highest concentrations in the late luteal phase and the lowest levels in the early follicular phase, which suggests that circulating free leptin is up-regulated by the C 21-steroid (P). Circulating bound leptin and soluble leptin receptor are not altered by the cyclic hormone status. The significant rise of the leptin bioequivalent, free leptin, in the late luteal phase might be of importance for the luteal-follicular and the luteal-preimplantatory functional shift.

Expanded clinical trial of emergency contraception with 10 mg mifepristone

We conducted a clinical single-arm trial to evaluate the effectiveness of 10 mg mifepristone for emergency contraception (EC) in a large population in China. The participating centers were 31 family-planning clinics and hospitals in the following 19 provinces or municipalities in China: Beijing, Shanghai, Tianjin, Harbin, Changchun, Shengyang, Shijiazhuang, Zhengzhou, Taiyuan, Nanjing, Jinan, Hangzhou, Guangzhou, Wuhan, Changsha, Chongqing, Guiyang, Chengdu, Kunming. A total of 4945 women requesting EC within 120 h after a single act of unprotected intercourse were recruited and treated with 10 mg mifepristone. A total of 28 women were lost to follow-up, and 4917 women were included in the analysis, of whom, 69 became pregnant. The combined pregnancy rate was 1.4 [95% confidence interval (CI): 1.0–1.9] and the percentage of pregnancies prevented was 82.2% (95% CI: 77.5–86.2%). There was a significant inverse trend in pregnancy rate with body mass index that disappeared when adjusted for other variables. The pregnancy risk was double among nulliparous women compared to parous women (2.3% compared to 1.0%), and it increased by a factor of 1.5 when the treatment was administered at 25–48 h and at 49–72 h compared to administration within 24 h, although this association was not significant. The risk of pregnancy was higher if intercourse took place during the follicular or preovulatory phase of the cycle. Women having repeated intercourse after treatment without any contraceptive methods had a dramatic increase in the risk of pregnancy, while those who used contraceptives had similar risk to those without acts. Side effects were mild and present in only small proportions of women: nausea and vomiting in 9% and other side effects in 2–3%. Delay of menstruation of 7 days or more occurred in 6.5% of women. The expanded study confirmed the high efficacy and safety of 10 mg mifepristone for EC.

Definition of the microvascular pattern of the normal human adult mammary gland.

The present report provides a detailed description of microvascularization in the normal human mammary gland, and defines two novel morphometric parameters to be used as a reference when angiogenesis in breast carcinoma is evaluated. Microvascularization was analysed by histology, immunohistochemistry and computer-assisted analysis in a set of breast tissue samples taken from nine women, during the pre-ovulatory phase of the menstrual cycle. The two parameters designed for image analysis were: vascular density (VD): [microvessel number/(microvessel area + residual stromal area)] x 10 000; and vascular area ratio (VAR): [microvessel area/(microvessel area + residual stromal area)]. In the lobules VD (mean value +/- SE 2.48 +/- 0.14) and VAR (0.33 +/- 0.02) showed little variability and correlated significantly (P < 0.05). The areas occupied by microvessels, stroma and acini remained constant in all lobules (21.53 +/- 1.87%, 42.65 +/- 1.35% and 35.14 +/- 1.57%, respectively). Microvascularization of the lobules was of a sinusoidal type, with large S-shaped capillaries. In the ducts VD (2.95 +/- 0.16) and VAR (0.29 +/- 0.03) showed little variability but did not correlate significantly. Microvascularization of the ducts was of a classic type, with capillaries normal in size and shape. The expression of oestrogen (ER) and progesterone (PR) receptors was analysed by immunohistochemistry and compared with the morphometric results. ER expression levels were in the range 20-25% (24.3 +/- 2.1) and 14-18% (15.4 +/- 1.5) in lobules and ducts, respectively. PR expression levels were in the range 10-13% (11.1 +/- 1.6) and 14-17% (15.2 +/- 1.4), respectively. No correlation was found between ER/PR expression and vascularization parameters.

Influence of endogenous oestrogens on QT interval duration

Women have a longer QT interval and a greater incidence of torsades de pointes than men. It has been suggested that oestrogens may influence the duration of cardiac repolarization. We thus investigated the influence of oestradiol (E2) on ventricular repolarization within the physiological concentration range of this hormone. We studied QT interval duration in 21 healthy women aged 18 to 35 years with regular menstrual cycle (mean duration: 29+/-1 days) during two periods associated with a wide range of oestradiol plasma levels: low level during menses (105+/-34 pmol/l) and high level during the pre-ovulatory phase (750+/-277 pmol/l). We used heart rate-independent assessment of QT. QT-RR pairs were measured over a wide range of RR intervals obtained at rest and during a sub-maximal exercise test. Using a monoexponential nonlinear curve fitting for the QT-RR relation, the QT(1000 ms)during nadir and peak oestradiol periods was then determined for each subject. QT(1000 ms)interval was not different between both study periods: 382.1+/-18.4 ms at peak versus 382.2+/-19.4 ms at nadir oestradiol level (P=0.98). No significant change in QT interval duration was observed within the large range of physiological E2 variations found during the menstrual cycle.