Biomedical subjects

F Raga

Publications and source records attributed to F Raga.

At least 19 recordsLinked to original sources

PubMed2001

Interleukin (IL)-1beta regulation of IL-1beta and IL-1 receptor antagonist expression in cultured human endometrial stromal cells.

The interleukin (IL)-1 system is a major regulator of local cellular interactions during embryonic implantation. Because IL-1beta and IL receptor antagonist (IL-1ra) are both expressed in human endometrium, we hypothesized that an appropriate ratio of IL-1beta to IL-1ra might favor the process of embryo implantation. Therefore, we investigated IL-1 regulation of the quantitative ratio of IL-1beta/IL-1ra messenger RNA (mRNA) expression in human endometrial stromal cells using quantitative competitive PCR, as well as intracellular protein expression after stromal cell solubilization. Confluent stromal cell cultures were stimulated with human IL-1beta (0-1000 IU/mL) for 24 h. After 24 h, total RNA was extracted, reverse transcribed, and coamplified by PCR with a defined amount of internal standard. The quantitative ratio was determined by the density of target to the internal standard. After culture with IL-1beta for 24 and 48 h, stromal cells were solubilized, and the intracellular protein levels of IL-1beta and IL-1ra were measured by enzyme-linked immunosorbent assay. The IL-1beta and IL-1ra mRNA were both up-regulated, and IL-1R tI mRNA was down-regulated, by IL-1beta in a dose-dependent manner. The quantitative ratio of IL-1beta to IL-1ra mRNA was constant with the presence of increasing concentrations of IL-1beta (1-1000 IU/mL). IL-1beta and IL-1ra protein was not detected in conditioned media of cultures before addition of IL-1beta. IL-1beta and IL-1ra protein levels increased with increasing amounts of IL-1beta after solubilization of stromal cells. The IL-1beta was detectable after 12 h of culture, in comparison with IL-1ra, which was detectable after 24 h of IL-1beta stimulation. These results suggest that IL-1 may play a crucial role in embryo-maternal interaction by regulating stromal cell expression of IL-1beta and IL-1ra, resulting in an appropriate ratio during the process of embryonic implantation.

Binding, Competitive↗

PubMed2000

Human oviductal gonadotropin-releasing hormone: possible implications in fertilization, early embryonic development, and implantation.

The oviduct is host to gametes and early embryos at a critical point in their lives. It is clear that the interactions of gametes/early embryo with the maternal oviduct in an autocrine and paracrine manner provide a microenvironment that enhances fertilization, early embryonic development, and implantation. Moreover, there is considerable evidence that an extrahypothalamic GnRH may play a substantial role as a molecular autocrine/paracrine regulator in these events. Gametes and preimplantation embryos express GnRH and GnRH receptor at both messenger ribonucleic acid (mRNA) and protein levels. However, whether GnRH is produced by the human oviduct has not yet been demonstrated. We used RT-PCR and immunohistochemical techniques to investigate GnRH mRNA and protein expression in human fallopian tubes throughout the menstrual cycle of premenopausal fertile patients. Our results, at both the mRNA and protein levels, revealed cycle-dependent production of an oviductal GnRH with expression during the luteal phase. Moreover, GnRH immunostaining was localized in the tubal epithelium during the luteal phase. On the basis of these data, we suggest that during reproductive life, oviductal GnRH may play a substantial paracrine/autocrine role in human fertilization, early embryonic development, and implantation.

Adult↗

PubMed1999

Assessment of endometrial volume by three-dimensional ultrasound prior to embryo transfer: clues to endometrial receptivity.

The aim of the present prospective study was to obtain quantitative data on endometrial volume by three-dimensional (3D) ultrasound at the time of embryo transfer in an in-vitro fertilization programme and to assess its value in predicting endometrial receptivity. The cycles (n = 72) were classified according to endometrial volume: group A <2 ml, group B 2-4 ml, and group C >4 ml. Comparisons of the groups showed that pregnancy and implantation rates were significantly lower (P < 0.05) in the group of patients with an endometrial volume <2 ml. Furthermore, no pregnancy was achieved with an endometrial volume <1 ml. It is concluded that endometrial volume by 3D transvaginal ultrasound may become a new objective parameter by which to predict endometrial receptivity.

Adult↗

PubMed1999

Recombinant follicle stimulating hormone stimulation in poor responders with normal basal concentrations of follicle stimulating hormone and oestradiol: improved reproductive outcome.

A total of 30 young infertile patients who exhibited a poor response in two previous consecutive cycles, despite having normal basal follicle stimulating hormone (FSH) and oestradiol concentrations, were invited to participate in a prospective randomized study comparing the clinical efficacy of recombinant (rFSH) and urinary (uFSH) follicle stimulating hormone. An evaluation of the total dose used (3800 IU versus 4600 IU, P < 0.05) and duration of treatment (10.2 days versus 13.2 days, P < 0.05) showed a significantly shorter treatment period as well as a significantly lower total dose of FSH required to induce ovulation successfully in the group of patients treated with rFSH. Significantly more oocytes (7.2 versus 5. 6, P < 0.05) as well as mature oocytes (5.9 versus 3.2, P < 0.01) were retrieved after rFSH treatment. In addition, significantly more good quality embryos were obtained (3.4 versus 1.8, P < 0.05) in the group of patients treated with rFSH and, as a result, higher pregnancy (33 versus 7%, P < 0.01) and implantation (16 versus 3%, P < 0.01) rates were achieved in these patients. It is concluded that rFSH is more effective than uFSH in inducing multifollicular development and achieving pregnancy in young low responders.

Adult↗

PubMed1999

GnRH mRNA and protein expression in human preimplantation embryos.

Gonadotrophin-releasing hormone (GnRH) regulates gonadotrophin biosynthesis and release in the anterior pituitary via specific receptors. Extrapituitary expression and action of GnRH have been demonstrated in several species. A possible role for GnRH in preimplantation embryonic development, endometrial preparation, and the implantation process has been previously suggested. Moreover, the presence of an immunoreactive GnRH in preimplantation embryos has been demonstrated in different species; however, there are no data for human embryos. We postulate that in humans GnRH may play a role in preimplantation embryonic development as well as in the implantation process. To examine this hypothesis, we assessed GnRH and GnRH-receptor mRNA and protein expression in human preimplantation embryos with three pronuclei. GnRH is expressed in peri-implantation human embryos at both the mRNA and protein level. GnRH-receptor mRNA is also present in the embryos studied. Immunohistochemical localization of GnRH showed intense staining in all the blastomeres at morula stage as well as in the trophectoderm and inner cell mass of the blastocysts. The results of the present study challenge the widely held view that GnRH has a predominantly central action, and suggests a pathway to describe a local role for the GnRH system in successful preimplantation embryonic development and implantation.

Actins↗

PubMed1999

Expression of mRNA for vascular endothelial growth factor transmembraneous receptors Flt1 and KDR, and the soluble receptor sflt in cycling human endometrium.

The aim of this study was to quantify and localize the mRNA expression of the vascular endothelial growth factor (VEGF) receptors Flt1, KDR and sflt, in human endometrium throughout the menstrual cycle. Since neoangiogenesis is crucial during embryonic implantation, we postulate that endometrial receptivity to VEGF may be altered during the luteal phase in order to support implantation. Human endometrium was collected and specified as early proliferative (n = 3), mid-proliferative (n = 4), late proliferative (n = 3), early secretory (n = 2), mid-secretory (n = 4), and late secretory (n = 4). Competitive reverse transcription-polymerase chain reaction (RT-PCR) was performed to evaluate the mRNA values throughout the menstrual cycle. Additionally, four samples were separated into epithelial and stromal-enriched cell fractions and competitive RT-PCR was carried out to specify the distribution of the mRNA expression. While mRNA for the transmembraneous receptors Flt1 and KDR was shown to be present at almost constant values throughout the menstrual cycle, the soluble receptor, sflt, had a three-fold higher level of transcription during mid-proliferative and late proliferative when compared with early proliferative and the entire secretory phase. The expression of Flt1, KDR and sflt mRNA was detected in both isolated endometrial epithelial and stromal cell fractions. In conclusion, the down-regulation of sflt, which functions as a soluble antagonist, during the luteal phase may act to sensitize the maternal endothelial receptors to angiogenetic stimuli secreted by the implanting embryo.

Adult↗

PubMed1999

The role of gonadotropin-releasing hormone in murine preimplantation embryonic development.

Previous studies have established the presence of an extrahypothalamic GnRH in a variety of tissues. GnRH receptor is known to be present in the placenta, which produces and secretes the decapeptide from the very early stages of placentation. We hypothesized that GnRH may play a role in the preimplantation development of embryos. To examine this hypothesis, we assessed GnRH and GnRH receptor messenger RNA (mRNA; RT-PCR) and protein expression (Immunohistochemistry) in preimplantation murine embryos at various developmental stages. Furthermore, preimplantation murine embryos were cultured with GnRH agonist and antagonist in vitro to assess the influence of GnRH analogs on embryo development. GnRH is expressed in the developing mouse embryo from morula to hatching blastocyst stages at the mRNA and protein levels. GnRH receptor mRNA is also present in the developing embryos studied. Preimplantation embryonic development was significantly enhanced by incubation with increasing concentrations of GnRH agonist and is significantly decreased by GnRH antagonist compared with that in the control group. Moreover, GnRH antagonist (5 and 10 microM) was able to completely block embryo development. The deleterious effect of GnRH antagonist on embryo development was reversed by increasing concentrations of the agonist, as determined by the number of embryos reaching the blastocyst stage.

Animals↗

PubMed1999

Independent regulation of matrix metalloproteinase-9, tissue inhibitor of metalloproteinase-1 (TIMP-1), and TIMP-3 in human endometrial stromal cells by gonadotropin-releasing hormone: implications in early human implantation.

Early human trophoblast shows dramatic invasive properties during early pregnancy. The simultaneous synthesis of matrix metalloproteinases (MMPs) and their specific tissue inhibitors (TIMPs) in both human trophoblast and decidual membranes suggests that their controlled and balanced expression is crucial for the rapid matrix remodeling and controlled invasion during early pregnancy. Recently, we have described the presence of an extrahypothalamic GnRH immunologically, biologically and chemically identical to the hypothalamic hormone in periimplantation human embryos. Moreover, the production of this decapeptide by the human trophoblast during the early stages of placentation is well documented. TIMP-1 and -3 messenger ribonucleic acid (mRNA) expression in cultured stromal cells and protein secretion into the medium were significantly decreased by GnRH agonist compared to that in control groups. Moreover, expression of TIMP-1 was affected to a greater extent than that of TIMP-3. GnRH antagonist ablated the down-regulation of TIMPs by the GnRH agonist. MMP-9 mRNA expression was not detected in the control groups or in the groups treated with GnRH analogs. Our results provide evidence that trophoblastic GnRH may play an important role in placental tissue organization and in the early embryo-maternal dialogue by enhancing trophoblast invasion through the specific inhibition of TIMPs.

Adult↗

PubMed1999

Polycystic ovarian syndrome: assessment with color Doppler angiography and three-dimensional ultrasonography.

We conducted a three-dimensional ultrasonographic evaluation of the size and structure of the ovaries of women who had clinical and biochemical findings suggestive of polycystic ovary syndrome. We carried out a comparative color Doppler frequency and color Doppler amplitude study of the vascular patterns of these ovaries. This study involved 65 women of reproductive age with polycystic ovary syndrome and 25 eumenorrheic women who were not taking hormonal contraceptives and who had a body mass index below 25 kg/m2. Compared to controls, women with polycystic ovary syndrome had larger ovaries and thicker stroma, increased impedance in the uterine arteries, increased stromal vascularity with decreased impedance that persisted throughout the menstrual cycle, and a lack of luteal conversion.

Adult↗

PubMed1998

Immunoreactive gonadotropin-releasing hormone expression in cycling human endometrium of fertile patients.

OBJECTIVE: To investigate the protein expression of GnRH in the endometrium of fertile patients throughout the menstrual cycle. DESIGN: Prospective longitudinal study. SETTING: Department of Gynecology and Obstetrics, Reproductive Immunology Laboratory, Stanford University Medical Center. PATIENT(S): Twenty-two fertile premenopausal women submitted to laparoscopic surgery for benign gynecologic indications. None of the 22 women had endometriosis or pelvic inflammatory disease. INTERVENTION(S): An endometrial biopsy specimen using the Novak curette was obtained at the time of surgery. MAIN OUTCOME MEASURE(S): Protein expression and localization from unfractioned endometrial tissue was analyzed by immunohistochemistry. RESULT(S): Gonadotropin-releasing hormone is expressed at the protein level in both the endometrial stroma and epithelium throughout the entire menstrual cycle of fertile women. Immunostaining in the human epithelium reached maximal levels in the midluteal phase and was elevated in the stroma throughout the entire luteal phase. CONCLUSION(S): Our results demonstrate the presence of GnRH in the human endometrium at the protein level throughout the entire menstrual cycle of fertile women, with an increase in the luteal phase compared with the preovulatory endometrium.

Adolescent↗

PubMed1998

Laparoscopic treatment of type IV rectovaginal fistula.

Fistulas between the anorectum and vagina may arise from several causes. Treatment depends on their etiology and location, as well as the surgeon's experience. Operative laparoscopy was successful in two women with type IV (mid)rectovaginal fistula in whom previous surgical attempts failed. Our experience suggests that mid and high rectovaginal fistulas can be effectively treated by laparoscopy in the hands of experienced endoscopic surgeons.

Adult↗

PubMed1998

Quantitative gonadotropin-releasing hormone gene expression and immunohistochemical localization in human endometrium throughout the menstrual cycle.

GnRH is one of the paracrine/autocrine regulators of hCG secretion produced by the human trophoblast during pregnancy. We hypothesized that GnRH may play a role in the embryonic/endometrial dialogue during early implantation. To examine this hypothesis, we assessed GnRH and GnRH-receptor mRNA and protein expression in human endometrium throughout the menstrual cycle of premenopausal fertile patients. Quantitation of the mRNA was performed by reverse transcription (RT)-competitive polymerase chain reaction (PCR) in the presence of a competitive cDNA fragment. RT-PCR revealed that unfractioned endometrium and isolated endometrial stromal and epithelial cells express GnRH and GnRH-receptor mRNA throughout all phases of the menstrual cycle. Quantitative PCR showed a dynamic pattern in the GnRH mRNA expression throughout the cycle, with a significant increase (p < 0.05) in the secretory phase as compared to the proliferative phase. Furthermore, quantitative competitive PCR of isolated glandular and stromal cells showed higher mRNA levels (p < 0.05) in the luteal phase in both compartments. GnRH immunostaining was localized in all major compartments, with the most intense staining during the luteal phase. On the basis of these data, we suggest that during reproductive life, endometrial GnRH may play a paracrine/autocrine role in the early stages of implantation by modulating embryonic trophoblastic secretion of hCG.

Adolescent↗

PubMed1998

First-trimester neck abnormalities: three-dimensional evaluation.

In order to study the first trimester ultrasonographic differences between nuchal translucency and hygroma colli, we rescanned 25 fetuses (13 with nuchal translucency and 12 with hygroma colli) using transvaginal and three-dimensional ultrasonography, after obtaining a fetal karyotype report. Our objective was to test the premise that the different physiopathologic mechanisms of both processes would be reflected in detectable sonographic differences. Our retrospective analysis showed that the most striking ultrasonographic difference was the presence of bullae as well as greater irregularity, extent, and amplitude of the membrane in cases of hygroma colli. Fetuses with simple nuchal translucency had a more homogeneous linear membrane. Although detailed analysis was impossible in 30% of cases, we found three-dimensional ultrasonography to be a useful technique for establishing the differences between these two entities.

Female↗

PubMed1998

Early diagnosis of conjoined twins using two-dimensional color Doppler and three-dimensional ultrasound.

Transvaginal three-dimensional (3-D) and color Doppler ultrasound were used to establish a first-trimester definitive diagnosis and classification of thoracoomphalopagus conjoined twins following two-dimensional (2-D) transabdominal and transvaginal scans that indicated twin gestation of uncertain classification. Color Doppler in combination with 3-D ultrasound can be a useful complement to 2-D ultrasound to confirm early diagnosis and determine the extent of organ sharing and definitive classification of conjoined twins.

Abortion, Induced↗

PubMed1997

Three-dimensional hysterosonography for the study of endometrial tumors: comparison with conventional transvaginal sonography, hysterosalpingography, and hysteroscopy.

We studied endometrial thickness and homogeneity in 36 patients with postmenopausal bleeding using three-dimensional ultrasound following distention of the uterine cavity with a sterile saline solution (3D-SHSG). Results with 3D-SHSG were compared with findings using transvaginal sonography, transvaginal sonohysterography, transvaginal color Doppler, and hysteroscopy. Sixteen patients (including three on tamoxifen) were undergoing hormone therapy at the time when they were studied. Visualization of the uterine cavity and of endometrial thickness was better with 3D-SHSG than with any of the other ultrasound techniques. The results with 3D-SHSG corresponded to the findings observed with hysteroscopy. Three-dimensional SHSG seems to improve ultrasound determination of myometrial and cervical invasion in cases of endometrial adenocarcinoma.

Endometrial Hyperplasia↗

PubMed1997

Reproductive impact of congenital Müllerian anomalies.

This retrospective longitudinal study was undertaken in order to determine the incidence and reproductive impact of uterine malformations on women desiring to conceive during their reproductive years. A total of 3181 patients in whom the morphology of the uterus was ascertained by hysterosalpingography (HSG) and laparoscopy/laparotomy during the years 1980-1995 was included in the study. The population analysed included fertile, infertile and sterile patients. The overall frequency of uterine malformations was 4.0%. Infertile patients (6.3%) had a significantly (P < 0.05) higher incidence of Müllerian anomalies, in comparison with fertile (3.8%) and sterile (2.4%) women. Septate (33.6%) and arcuate (32.8%) uteri were the most common malformations observed. Each malformation was individually analysed in fertile and infertile patients, in order to ascertain its actual reproductive impact. The performance of the unicornuate and didelphys uteri was similar with a chance of having a living child of 37-40%. The reproductive potential of the bicornuate uterus showed a live birth rate of 62.5% and the septate uterus showed a live birth rate of 62%. In all these abnormalities, early miscarriages (25-38%) and preterm deliveries (25-47%) were quite common. The arcuate uterus presented a live birth rate of 82.7%. It is concluded that uterine anomalies are relatively frequent in fertile women, and more frequent in infertile patients. Nevertheless, fertile patients with normal reproductive performance do exist, and Müllerian defects can permit an absolutely normal obstetric outcome. The reproductive performance of the unicornuate and didelphys uteri was poor, while that of the septate and bicornuate uteri was better than expected. The arcuate uterus had no impact on reproduction.

Abortion, Spontaneous↗

PubMed1996

Control of intrauterine device insertion with three-dimensional ultrasound: is it the future?

A comparative study was designed to determine whether three-dimensional transvaginal sonography (3D-TVS) offered advantages over two-dimensional transvaginal sonography (2D-TVS) for the identification and location of IUDs in 66 asymptomatic women. Hysteroscopy was performed in cases in which there was a discrepancy between the information obtained by both methods (n = 14). In eight cases (12.2%) the IUD was misidentified with 2D-TVS. In six cases (9.1%) it was not possible to identify the device model with 2D-TVS. In two cases (3.0%) 2D-TVS failed to identify the position of the device. In contrast, all IUDs were identified and located accurately with 3D-TVS.

Adult↗

PubMed1996

Congenital Müllerian anomalies: diagnostic accuracy of three-dimensional ultrasound.

OBJECTIVE: To determine whether it is possible to identify and diagnose accurately Müllerian anomalies with three-dimensional (3-D) ultrasound (US). DESIGN: Controlled blinded clinical study. SETTING: Normal human volunteers undergoing infertility evaluation in a university hospital. PATIENTS: Forty-two patients who underwent laparoscopy and hysterosalpingography as part of their work up for infertility and were found to have either a normal uterus (30 patients) or a Müllerian abnormality (12 patients) consented to be evaluated with 3-D US by sonographers who were unaware of their infertility history or of their laparoscopy and hysterosalpingography diagnoses. INTERVENTIONS: Transvaginal 3-D US evaluation over a 10- to 15-minute duration. MAIN OUTCOME MEASURES: Three-dimensional imaging was successful in all cases. RESULTS: Sonographers identified a Müllerian anomaly in all cases and came up with the correct classification in 11 of 12 cases. All patients with a normal uterus were identified correctly. CONCLUSIONS: In all patients with Müllerian anomalies, 3-D US examination of the endometrial cavity correlated with hysterosalpingography. In 91.6% of patients, 3-D US correlated with the external uterine configuration observed by laparoscopy. This technique may be used reliably in an office setting to diagnose and classify Müllerian anomalies.

Adult↗