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J R Loret de Mola

Publications and source records attributed to J R Loret de Mola.

At least 19 recordsLinked to original sources
PubMed2004

Comparison of two culture systems for the in-vitro growth and maturation of mouse preantral follicles.

The objective of our study was to compare to ability of collagen-treated membranes and bovine collagen gels to maintain murine preantral follicle growth and development in-vitro. To fulfill that objective, murine follicle and oocyte growth rates were followed for ten days in culture. Meiotic competence and the capacity to reach the two-cell stage after in-vitro maturation and fertilization, respectively, were then assessed. We used preantral follicles from 12 day-old CF-1 female mice that were isolated by enzymatic digestion from ovaries. Follicles were placed either on collagen-treated membranes or embedded in a bovine collagen matrix. The follicles were grown, changing the media and obtaining measurements every other day for ten days. Following culture, the granulosa-oocyte complexes were matured; the resultant metaphase II arrested oocytes were inseminated and cultured to the two-cell stage. The data was analyzed with significance considered for probability values of p < 0.05. We performed individual measurements on 650 follicles in seven separate experiments. Forty-eight hours after initial seeding and throughout the entire length of culture, both the follicles and oocytes grown in the collagen matrix were larger than follicles cultured on collagen-treated membranes (p < .0001). However, oocyte recovery rates were higher among follicles cultured on collagen-treated membranes (p < .01). Similar percentages of meiotically competent oocytes, fertilization and cleavage rates were observed in both groups. Our results show that mouse preantral follicles display a greater growth rate when grown embedded in a collagen gel matrix. This may be due to the maintenance of a normal three-dimensional organization of the follicle within the collagen matrix. However, this system does not enhance meiotic competency or fertilization rates in the mouse when compared to culture on collagen-treated membranes.

Animals↗
PubMed2002

Treatment of murine breast cancer cells with antisense RNA to the type I insulin-like growth factor receptor decreases the level of plasminogen activator transcripts, inhibits cell growth in vitro, and reduces tumorigenesis in vivo.

AIMS: To establish that cells from the murine mammary carcinoma cell line, EMT6, express type I insulin-like growth factor receptor (IGF-IR), tissue-type plasminogen activator (tPA), and urokinase-type plasminogen activator (uPA). To investigate the role of IGF-IR in growth, transformation, and tumorigenesis in addition to its relation to tPA and uPA in EMT6 cells. To assess the suitability of the EMT6/syngeneic mouse model for studying the role of IGF-IR in tumorigenesis. METHODS: The presence of transcripts for IGF-IR, tPA, and uPA was determined by northern blot analysis using poly (A(+)) RNA derived from EMT6 cells transfected with an antisense IGF-IR construct or a construct lacking the antisense IGF-IR insert. Flow cytometry was used to measure IGF-IR protein. Assays were performed to determine cell proliferation, transformation, and the tumorigenicity of antisense IGF-IR transfected EMT6 cells and control transfected EMT6 cells. RESULTS: There was strong expression of IGF-IR, tPA, and uPA in EMT6 cells. EMT6 cells from clones carrying antisense IGF-IR displayed a significant decrease in cell proliferation and lost the ability to form colonies in soft agar. A decrease in tumour size occurred when cells carrying the antisense IGF-IR were injected into syngeneic mice. Reduced expression of tPA and uPA was seen in EMT6 cells carrying the antisense IGF-IR construct. CONCLUSIONS: The IGF-IR plays a role in the progression, transformation, and tumorigenesis of EMT6 murine mammary carcinoma cells. The suppression of IGF-IR mRNA in EMT6 cells decreases tPA and uPA expression. EMT6 cells and the syngeneic mouse provide a suitable model for studying the role of IGF-IR in breast tumour progression.

Animals↗
PubMed2000

Subclavian deep vein thrombosis associated with the use of recombinant follicle-stimulating hormone (Gonal-F) complicating mild ovarian hyperstimulation syndrome.

OBJECTIVE: To describe two cases of subclavian deep vein thrombosis (DVT) associated with the use of recombinant gonadotropins and mild ovarian hyperstimulation syndrome (OHSS) and review the literature associated with this condition. DESIGN: Retrospective study (case report). SETTING: Tertiary academic IVF program. PATIENT(S): Two women undergoing IVF with intracytoplasmic sperm injection due to male factor infertility. INTERVENTION(S): Ovaluation induction with recombinant FSH, IVF, and therapeutic heparinization. MAIN OUTCOME MEASURE(S): Coagulation studies, resolution of DVT, delivery at term. RESULT(S): Mild OHSS with left subclavian thrombosis occurred in two patients. Laboratory evaluation revealed normal protein C, protein S, antinuclear antibodies (ANA), and absence of antiphospholipid (APA) and anticardiolipin antibodies (ACA). Antithrombin III levels and coagulation studies were also within normal limits. Both patients tested negative for a factor V Leiden mutation and delivered healthy infants at term. CONCLUSION(S): Arm swelling associated with the use of gonadotropins during controlled ovarian hyperstimulation should be promptly evaluated and treated. Subclavian vein thrombosis is a rare complication of ovulation induction, and the possibility that recombinant gonadotropins increase the risk for this complication should be further studied.

Adult↗
PubMed1999

Gonadotropins induce higher active renin levels in the follicular fluid of normal and hyperstimulated cycles.

The objective of this study was to analyze follicular fluid active renin and its relationship to steroid hormones throughout the normal and gonadotropin-stimulated menstrual cycle. Active renin was measured in the follicular fluid of patients undergoing tubal sterilization (n = 16) and in vitro fertilization (IVF) (n = 25); IVF patients were either in a natural cycle (n = 7) or undergoing controlled ovarian hyperstimulation (n = 18). The largest visible follicle was aspirated at the time of laparoscopic tubal sterilization; ultrasound guided transvaginal follicular aspiration was used in the IVF group. Follicular fluid active renin, estradiol and progesterone levels were measured with immunoradiometric and fluoroimmunoassays. The cycle day was correlated with the spontaneous luteinizing hormone (LH) surge or human chorionic gonadotropin (hCG) administration, as well as active renin, estradiol, progesterone levels and the estradiol/progesterone ratio using simple and multiple regression and analysis of variance (ANOVA). Cycle day independently influenced active renin, progesterone and the estradiol/progesterone ratio in a statistically significant manner (p < 0.0001). The active renin and progesterone levels were highest during the periovulatory period (p < 0.0001 and p < 0.002, respectively) and the estradiol/progesterone ratio correlated inversely with cycle day (p < 0.003). Although the follicular fluid active renin, estradiol and progesterone levels were higher after controlled ovarian hyperstimulation when compared to natural cycles, this difference did not reach statistical significance. Our findings suggest that active renin levels in follicular fluid increase in the follicular phase of the menstrual cycle, reaching peak levels in the periovulatory period following the LH surge or hCG administration, providing indirect support for the hypothesis that the ovarian renin-angiotensin system (RAS) is under gonadotropin control.

Adult↗
PubMed1998

Identification of oncofetal fibronectin in patients with advanced epithelial ovarian cancer: detection in ascitic fluid and localization to primary sites and metastatic implants.

BACKGROUND: The mechanisms by which metastatic ovarian cancer adheres to peritoneal surfaces are not well understood. A role for tumor-derived extracellular matrix adhesive molecules such as fibronectin (FN) has been proposed. Because oncofetal fibronectin (onfFN) isoforms function in the adhesion of trophoblasts and have been identified in association with several malignancies, we sought to study onfFN in patients with advanced epithelial ovarian cancer. METHODS: Total FN was identified with the nonspecific anti-FN monoclonal antibody CAF. OnfFN was identified using the specific monoclonal antibodies FDC-6 and X18A4. These antibodies were applied to: 1) ascitic fluid from advanced epithelial ovarian cancer patients and peritoneal fluid from patients without pathologic conditions and 2) tissue sections of primary lesions and metastatic ovarian cancer implants. Comparative histologic specimens included normal ovarian tissue and small bowel implants of endometriosis. RESULTS: When measured by sandwich enzyme-linked immunoadsorbent assay, all peritoneal fluids (32 malignant and 32 benign) contained marked quantities of total (CAF reactive) FN, although malignant ascites had higher concentrations than benign samples (173.2 +/- 36.8 microg/mL vs. 76.4 +/- 31.8 microg/mL; P = 0.001). Malignant ascites also had significantly higher levels of onfFN than benign peritoneal fluid (FDC-6: 3.4 +/- 0.6 vs. 0.9 +/- 0.2 microg/mL; and X18A4: 5.1 +/- 1.3 vs. 1.1 +/- 0.4 microg/mL; P = 0.0001). Immunohistochemical staining of malignant lesions revealed prominent localization of both CAF reactive FN and onfFN to the stroma surrounding epithelial tumor nests. More delicate fibrillar staining within tumor nests also was evident. In contrast, implants of endometriosis revealed strong stromal staining for CAF reactive FN but not for onfFN. CONCLUSIONS: These results demonstrate the presence of onfFN in advanced ovarian malignancies. We speculate that onfFN may participate in tumor-associated peritoneal adhesive interactions.

Adenocarcinoma, Clear Cell↗
PubMed1998

Reliability of intraobserver and interobserver sonographic endometrial stripe thickness measurements.

OBJECTIVE: To determine the intraobserver and interobserver reliability of endometrial stripe thickness measurements in women undergoing controlled ovarian hyperstimulation. DESIGN: Prospective blinded study. SETTING: Tertiary care, university hospital. PATIENT(S): Sixty-three patients undergoing controlled ovarian hyperstimulation and being monitored with transvaginal ultrasound were studied. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Intraobserver and interobserver variability of endometrial stripe thickness measurements between 2 blinded observers, with 2 observations made by each observer. RESULT(S): A statistically significant correlation was detected between the 2 measurements of each observer. The mean (+/-SD) interobserver difference between the average of the 2 measurements performed by both observers was 1.02 +/- 0.82 mm. A statistically significant correlation between the measurements of the 2 observers was detected. For both observers A and B, who used < or =6 mm as an abnormal endometrial thickness, an excellent level of agreement was detected between the 2 measurements made on each patient. When comparing the average values obtained by the 2 observers for each of the patients, an excellent level of agreement was detected. CONCLUSION(S): There is an excellent correlation between intraobserver and interobserver measurements of endometrial stripe thickness.

Adult↗
PubMed1998

Gonadotropins induce the release of interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha from the human preovulatory follicle.

PROBLEM: The effects of exogenous gonadotropin administration and steroid levels on the release of various cytokines into the human follicular fluid (FF) were studied. METHOD OF STUDY: Forty patients were included in two groups, those undergoing controlled ovarian hyperstimulation (COH) (n = 33) and natural cycles (n = 7). FF transvaginal aspirations were performed 36 hr after administration of human chorionic gonadotropin or a spontaneous surge of luteinizing hormone, respectively. FF cytokine measurements were performed with sensitive immunoassays. RESULTS: FF cytokine levels were higher after COH [interleukin (IL)-1 beta, 6.6 +/- 0.32 pg/ml; IL-6, 18.7 +/- 2.1 pg/ml; and tumor necrosis factor (TNF)-alpha, 32.5 +/- 4.9 pg/ml] than in natural unstimulated cycles (0.52 +/- 0.1 pg/ml, P < 0.001; 8.9 +/- 1.2 pg/ml, P < 0.01; and 13.2 +/- 2.6 pg/ml, P < 0.001, respectively). FF estradiol (E2) and progesterone levels were not statistically different between groups, despite the higher serum E2 levels observed in patients after COH. CONCLUSIONS: Gonadotropins might regulate ovarian secretion of cytokines, because FF IL-1 beta, IL-6, and TNF-alpha levels after COH were higher than during natural cycles.

Adult↗
PubMed1998

Indirect prediction of fetal lung maturity. Value of ultrasonographic colonic and placental grading.

OBJECTIVE: To correlate fetal morphometrics with studies of fetal lung maturity. STUDY DESIGN: One hundred six patients undergoing amniocentesis for fetal lung maturity studies were examined prospectively. Eighty-four patients were normal (79%), and 22 were diabetic (21%). Fetal morphometrics were obtained prior to amniocentesis. The fetal colon and placenta were graded. Discriminant analysis was used to identify variables that were predictive of a mature lecithin/sphingomyelin ratio and the presence of phosphatidylglycerol (PG). All patients delivered within 48 hours of amniocentesis. RESULTS: In the normal group, 28 (33%) fetuses had a grade 3 colon, which was 68% sensitive and 98% specific for a mature amniocentesis. A grade 3 colon was the single best predictor of a mature amniocentesis (P < .001). Twenty-five (29%) fetuses had a grade 3 placenta, which was 64% sensitive and 96% specific for a mature amniocentesis (P < .005). Diabetes did not influence colonic grading since a grade 3 colon was present in seven (32%) patients (47% sensitivity and 100% specificity for PG) (P < .02). Interexaminer and intraexaminer variability for the study was excellent, kappa = 1.0 (P < .001). CONCLUSION: Colonic and placental stage 3 grading are reliable and reproducible ultrasonographic scales that can help predict the findings of fetal lung maturity studies.

Adult↗
PubMed1997

Sequential analysis of zona thickness during in vitro culture of human zygotes: correlation with embryo quality, age, and implantation.

Zona pellucida thickness was measured daily in zygotes and cleavage stage embryos. Measurements were performed on a Nikon inverted microscope equipped with Hoffman modulation optics, using an ocular micrometer. Zona thickness of each zygote/embryo was measured four times, the zygote/embryo was then "rolled over," and four more measurements were repeated for a total of eight. The zygotes/embryos were photographed daily and the measurements repeated on the prints. Subsequently, the mean zona thickness for each stage was calculated. A total of 81 patients (mean age 33.8 +/- 4.2) participated in the study. A total of 427 embryos were evaluated. Categorical data differences between groups were evaluated by ANOVA and multiple linear regression. For nominal data, the Kruskal-Wallis test was applied; when P < 0.05 the differences were considered to be significant. We found that the average zona thickness on day 1 of in vitro culture was 17.7 +/- 0.14 microns; 16.3 +/- 0.14 microns on day 2 and 14.9 +/- 0.14 microns on day 3 (P < .0001). When the zona thickness was analyzed in relation to the number of blastomeres on day 3 of culture, there was a highly significant correlation with blastomere number (P < .0001). Similarly, there was a highly significant correlation with embryo grade (P < .005) and fragmentation (P < .001). The data were also analyzed for embryos transferred that resulted in a successful pregnancy, revealing that embryos in a pregnancy cycle had significantly thinner zonae pellucidae (P < .0001), when compared to embryos that were not transferred or from nonconceptual cycles. The average zona thickness also decreased with age, and was most apparent after 35 years. Changes in zona thickness correlated with the number of blastomeres, grade, fragmentation, age and were more evident in embryos transferred from cycles resulting in successful pregnancies. Therefore, zona pellucida measurements should be included in the overall assessment of embryo quality, since this information may be useful in the selection of optimal embryos for transfer.

Adult↗
PubMed1997

Markedly elevated cytokines in pleural effusion during the ovarian hyperstimulation syndrome: transudate or ascites?

OBJECTIVE: To study levels of proinflammatory cytokines in pleural fluid during the severe ovarian hyperstimulation syndrome (OHSS). DESIGN: Case report. SETTING: Tertiary academic medical center. PATIENT(S): A 35-year-old female with a 6-year history of unexplained infertility on menotropin therapy and 28 healthy normal controls. INTERVENTION(S): Thoracentesis for severe pleural effusion and venipunctures. MAIN OUTCOME MEASURE(S): Interleukin-1 beta (IL-beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha) levels were measured by ELISA and compared between pleural effusion and serum from normal controls. RESULT(S): Pleural effusion IL-1 beta and IL-6 levels were higher than serum. Interleukin-6 levels were elevated particularly in pleural effusion (1,961.89 pg/mL) compared with serum (3.9 +/- 0.41 pg/mL). CONCLUSION(S): Our results confirm the high cytokine levels observed in OHSS. Cytokines have been implicated in capillary permeability, extravasation of fluid, oliguria, and shock. We have postulated that these mediators are released from the corpora lutea into the peritoneum and systemic circulation. Alternatively, the presence of high cytokine levels in pleural fluid maybe the result of diaphragmatic defects, which allow for the migration of ascites into the pleural space.

Adult↗
PubMed1997

Hydrosalpinx fluid enhances human trophoblast viability and function in vitro: implications for embryonic implantation in assisted reproduction.

OBJECTIVE: To assess the effects of hydrosalpinx fluid on human cytotrophoblast viability and function in vitro. DESIGN: Human cytotrophoblasts obtained from third-trimester placentas were cultured in vitro with hydrosalpinx fluid, and cell viability and protein production were assayed. SETTING: A university hospital. PATIENT(S): Ten hydrosalpinx fluid samples obtained from seven women with clearly diagnosed hydrosalpinges. INTERVENTION(S): Recovery of hydrosalpinx fluid by transvaginal aspiration or at the time of surgery. MAIN OUTCOME MEASURE(S): Cell viability was assessed by the XTT assay. Secretion of trophoblast oncofetal fibronectin (tropho-uteronectin) and beta-hCG by cultured trophoblasts was determined by Western blot and ELISA of the culture media. RESULT(S): With increasing concentrations of hydrosalpinx fluid from 0% to 20%, there was a significant increase in trophoblast cell viability (1.63-fold increase in 20% hydrosalpinx fluid). Likewise, both Western blot and ELISA assays demonstrated a significant increase in tropho-uteronectin production by trophoblasts with increasing hydrosalpinx fluid concentrations (3.76-fold increase in 20% hydrosalpinx fluid). beta-Human chorionic gonadotropin production also increased significantly in the presence of hydrosalpinx fluid (3.31-fold increase in 20% hydrosalpinx fluid). CONCLUSION(S): These findings suggest that hydrosalpinx fluid improves human trophoblast viability in vitro and enhances the production of tropho-uteronectin and beta-hCG by these cells.

Adult↗
PubMed1997

Uterine arteriovenous malformation in a patient with recurrent pregnancy loss and a bicornuate uterus. A case report.

BACKGROUND: Arteriovenous malformations (AVM) of the uterus are a rare but potential cause of recurrent pregnancy loss. Only four cases of uterine AVM have resulted in a live birth after conservative management. There is no previous report in which a combination of a müllerian anomaly and an AVM existed concomitantly. CASE: A 33-year-old woman with a history of recurrent pregnancy loss was found to have a coexistent uterine AVM and a bicornuate uterus. The patient underwent arterial embolization and Strassman metroplasty and subsequently had a term pregnancy with a live birth. CONCLUSION: Management of uterine AVM should be individualized, taking into account the patient's desire for future fertility and the stability of her health at presentation.

Abortion, Habitual↗
PubMed1997

Endometrial changes with chronic tamoxifen use.

Tamoxifen is a nonsteroidal antiestrogenic drug used successfully to reduce recurrences in all stages of breast carcinoma. Recent reports have emphasized an increased incidence of premalignant and neoplastic endometrial changes that have questioned the risk-benefit ratio of this medication. It appears that the use of tamoxifen increases the incidence of endometrial abnormalities in 1.2 individuals per 1000 women treated with tamoxifen, increasing to 6.3 per 1000 women after 5 years. However, current evidence suggests that the use of tamoxifen as an adjuvant for breast carcinoma overrides the risk of developing endometrial neoplasias. It is not clear which long-term method of surveillance is most cost-effective for these patients. Therefore, individuals should be alerted to the potential risks of tamoxifen use. At present, the screening method of choice should be decided by each individual practitioner, bearing in mind that any vaginal bleeding should be readily investigated. If high-grade hyperplasias of the endometrium or cancer are found, management should be tailored in conjunction with the physician responsible for the woman's breast care.

Breast Neoplasms↗
PubMed1996

Elevated interleukin-6 levels in the ovarian hyperstimulation syndrome: ovarian immunohistochemical localization of interleukin-6 signal.

OBJECTIVE: To examine the production and immunolocalization of interleukin-6 (IL-6) in patients with the ovarian hyperstimulation syndrome. METHODS: The study group consisted of patients with ovarian hyperstimulation syndrome (n = 9) from whom serum and ascites samples were obtained. The control samples used were serum (n = 10) and peritoneal (n = 16) and follicular fluids (n = 8) from healthy individuals. Follicular fluid (n = 40) and serial serum samples were also obtained from patients undergoing menotropin stimulation for in vitro fertilization (IVF) before (n = 10) and after ovulation (n = 34). Interleukin-6 measurements were performed with a sensitive immunoassay and confirmed with a bioassay. Immunohistochemical localization of IL-6 was performed with a mouse monoclonal antibody in normal premenopausal (n = 5) and postmenopausal ovaries (n = 5), as well as with cells from stimulated follicular fluid aspirates (n = 3). RESULTS: We found significantly higher serum and ascites IL-6 levels in ovarian hyperstimulation syndrome (mean 18.8 +/- 1.1 and 810.8 +/- 60.7 pg/mL, respectively) compared with postovulatory serum and peritoneal fluid from normal controls (mean 4.4 +/- .69 and 44.7 +/- 7.5 pg/mL, respectively) (P < .001) or serum after menotropin stimulation (13.1 +/- 1.1 pg/mL) (P < .001). At the time of ovulation, follicular fluid IL-6 levels (normal controls, mean 9 +/- 2.1 pg/mL; menotropin stimulation, mean 10.1 +/- 4 pg/mL) were higher than in preovulatory serum (normal controls, mean 4.5 +/- .8 pg/mL; menotropin stimulation, mean 6.3 +/- 1.4 pg/mL) (P < .001). Immunohistochemical localization of IL-6 revealed intense staining in corpora lutea and theca cells from large antral follicles and luteinized granulosa cells in follicular aspirates after menotropin stimulation. CONCLUSION: Interleukin-6 levels are markedly elevated in the ovarian hyperstimulation syndrome when compared with controls. The higher follicular fluid IL-6 levels seen suggest local secretion of this cytokine. Immunohistochemical correlation demonstrated IL-6 within ovarian theca cells. These findings suggest a local role for IL-6 both in normal and stimulated ovarian function. Whether IL-6 is directly responsible for the clinical manifestations of this syndrome is unclear. However, when produced in massive amounts, the pro-inflammatory effects of IL-6 may contribute to its pathogenesis and perhaps serve as a marker for the disease.

Adult↗
PubMed1996

Ovarian hyperstimulation syndrome: pre-ovulatory serum concentrations of interleukin-6, interleukin-1 receptor antagonist and tumour necrosis factor-alpha cannot predict its occurrence.

The pathogenesis of the ovarian hyperstimulation syndrome (OHSS) is poorly understood. Since significant elevations in cytokines are found in OHSS, our objective was to conduct a prospective case-controlled study to assess if pre-ovulatory cytokine serum concentrations can predict its occurrence. The study group was selected from in-vitro fertilization patients who subsequently developed severe OHSS, along with a matched group who did not develop this complication (n = 20), and a healthy normal control group (n = 10). Interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1RA) and tumour necrosis factor-alpha (TNFalpha) measurements were performed with sensitive immunoassays and confirmed with bioassays. Serum IL-6 (mean concentrations +/- SEM: 4. 38 +/- 0.36 pg/ml), IL-1RA (829 +/- 292 pg/ml) and TNFalpha (15.5 +/- 1.32 pg/ml) concentrations did not show differences throughout the normal menstrual cycle group. Cytokine variability and pre-ovulatory values were similar in OHSS compared to controlled ovarian hyperstimulation (COH) patients. However, average follicular phase serum IL-6 concentrations were higher in OHSS (8.71 +/- 0.41 pg/ml) and COH (7.66 +/- 0.38 pg/ml) patients than in normally menstruating women (4.34 +/- 0.99 pg/ml) (P < 0.0001). Pre-ovulatory serum IL-6 concentrations were also higher in OHSS (9 +/- 0.94 pg/ml) and COH (7.3 +/- 0.97 pg/ml) patients than in controls (4.57 +/- 1.1 pg/ml) (P < 0.01 and P < 0.04 respectively). All IL-1RA and TNFalpha concentrations were comparable in all the groups. This study suggests that cytokine measurements cannot be used to predict the occurrence of OHSS prior to the administration of human chorionic gonadotrophin.

Adult↗