Carlos Sueldo

Laparoscopic ovarian cystectomy of endometriomas does not affect the ovarian response to gonadotropin stimulation

OBJECTIVE To evaluate the ovarian response cycles of IVF-ET in patients who previously underwent laparoscopic cystectomy for endometriomas. DESIGN Retrospective study with prospective selection of participants and controls. SETTING Instituto de Ginecología y Fertilidad Buenos Aires, Argentina. PATIENT(S) Thirty-nine patients underwent an operation for ovarian endometriomas by atraumatic removal of the pseudocapsule with minimal bipolar cauterization of small bleeders and an IVF-ET cycle (group A) and 39 control patients of similar age underwent an IVF-ET cycle for tubal factor infertility (group B). INTERVENTION(S) Laparoscopic endometrioma cystectomy, IVF-ET cycle. MAIN OUTCOME MEASURE(S) E(2) levels, number of gonadotropin ampoules, follicles, oocytes retrieved, number and quality of embryos transferred, and clinical pregnancy rate. RESULT(S) There were no differences in all the parameters studied (E(2) levels, number of follicles, oocytes retrieved, number and quality of embryos transferred, and clinical pregnancy rate) except for the number of gonadotropin ampoules needed for ovarian hyperstimulation, which was significantly higher in group A than in group B. CONCLUSION(S) Our results indicate that laparoscopic cystectomy for endometriomas is an appropriate treatment since it did not negatively affect the ovarian response for IVF-ET.

Oral contraceptives suppress cell proliferation and enhance apoptosis of eutopic endometrial tissue from patients with endometriosis

OBJECTIVE To evaluate the effects of administering combination oral contraceptives (COCs) to patients with endometriosis on the regulation of cell growth in the eutopic endometrium. DESIGN Prospective study. SETTING Research institute and clinical fertility center. PATIENT(S) Thirteen women with untreated endometriosis and 13 controls. INTERVENTION(S) Biopsy specimens of the eutopic endometrium were obtained from all subjects. Apoptosis, cell proliferation, and Bcl-2 and Bax expression were examined at the epithelial and stromal levels in the eutopic endometrium from patients with endometriosis before and after 30 days of daily exposure to COCs and from controls. MAIN OUTCOME MEASURE(S) Apoptotic cells were detected by using the dUTP nick-end labeling assay; Ki-67, Bcl-2, and Bax expressions were assessed by using immunohistochemical techniques. RESULT(S) After exposure to COCs, apoptosis was significantly increased in the eutopic endometrium compared with before COC administration, both at epithelial and stromal levels. Cell proliferation was significantly lowered by COCs. CONCLUSION(S) COCs showed a positive effect on patients with endometriosis by down-regulating cell proliferation and enhancing apoptosis in the eutopic endometrium.

Gonadotropin-releasing hormone agonist induces apoptosis and reduces cell proliferation in eutopic endometrial cultures from women with endometriosis

OBJECTIVE There is growing evidence that suggests a direct action of gonadotropin-releasing hormone agonist (GnRH-a) on endometrial growth. Consequently, our purpose was to evaluate the effect of GnRH-a on in vitro eutopic endometrial cell growth and apoptosis. DESIGN Prospective study. SETTING Research institute and clinical fertility center. PATIENT(S) Sixteen women with untreated endometriosis and 14 controls. INTERVENTION(S) Biopsy specimens of eutopic endometrium were obtained from all subjects. Apoptosis and cell proliferation were examined in epithelial endometrial cell cultures after incubation with leuprolide acetate (LA), antide, and a combination of both. MAIN OUTCOME MEASURE(S) The percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique; cell proliferation was assessed by (3)H-thymidine incorporation. RESULT(S) Leuprolide acetate (LA) (100 ng/mL) enhanced apoptosis in endometrial cultures from patients with endometriosis and controls, and this effect was reversed by antide 10(-7)M. Cell proliferation was down-regulated by LA at 1, 10, and 100 ng/mL in cultures from women without and with endometriosis. The addition of antide 10(-7)M reversed this inhibition. CONCLUSION(S) GnRH-a appears to have a direct effect by enhancing the apoptotic index and decreasing the cell proliferation in endometrial cells.

Reduction of primary postoperative adhesion formation under calcium channel blockade in the rabbit

Preliminary studies in a hamster model have demonstrated calcium channel blocking agents to be potent inhibitors of primary post-traumatic peritoneal adhesion formation. The present investigation was designed to extend these observations to an extensively studied model, the rabbit, and to evaluate the optimal route of administration of these drugs for intraabdominal surgery. Rabbits were subjected to a standardized traumatic lesion of the left uterine horn. Subsequently, animals were divided into the following treatment groups: subcutaneous vehicle control (n = 7), intraperitoneal (ip) vehicle control (n = 8), subcutaneous verapamil treatment (n = 6), low-dose (2.5 micrograms/kg/hr) ip verapamil treatment (n = 10), and high-dose ip (25 micrograms/kg/hr) verapamil treatment (n = 6). All animals were reexplored at 1 week postop for evaluation of adhesion formation (scale: 0 to 4+). Calcium channel blockade-treated animals formed significantly fewer adhesions (0.45) than controls (3.93) (P less than 0.01). There was no significant difference between animals treated with sc with verapamil sc and those treated with low- or high-dose verapamil ip (0.33 vs 0.20 vs 1.0). These data confirm our preliminary results, suggesting that calcium channel blockade potently modulates peritoneal healing and regeneration. Furthermore, intraperitoneal delivery and systemic administration appear equipotent in this model. Further study of these agents as potential adjuvants for intraperitoneal surgery is indicated.

Endometrial lesions caused by catheters used for embryo transfers: a preliminary report

OBJECTIVE To visualize by microhysteroscopy any possible lesions on the endocervix and endometrium made by the catheters commonly used for embryo transfer (ET). DESIGN Prospective descriptive study. Tertiary fertility center (IFER). PATIENT(S) Twenty-three infertile patients underwent a mock transfer before a microhysteroscopy during the postovulatory phase (days 2-5 after ovulation) of the cycle with a Tomcat catheter (n = 5), Frydmans catheter (n = 5), Frydmans set (n = 3), or Wallaces catheter (n = 10). INTERVENTION(S) Mock ETs and subsequent mycrohysteroscopies.Visualization, description, and documentation of endocervical and endometrial lesions. RESULT(S) The lesions in all 23 patients were described and documented (tunnel-like, groove-like, punch-out, crater-like). The Wallace catheter appears to be less traumatic to the endometrium (but it seems that it is important to take care to not pass the internal os with the outer sheath). The Tomcat catheter and the Frydmans set caused the more significant lesions that were observed. CONCLUSION(S) In this preliminary study, for the first time endometrial lesions caused by the ET catheters were directly visualized and documented. Some of these observed lesions appear to be capable of compromising the success of ET.

Correlation of semen transferrin concentration and sperm fertilizing capacity.

To determine whether a correlation exists between semen transferrin and sperm fertilizing capacity, transferrin concentration was determined in the semen of 52 male patients referred for the hamster ova penetration test (group 1) and in 17 men participating in the human in vitro fertilization program (group 2). In both groups 1 and 2 seminal transferrin levels were also compared to semen volume, sperm density, and motility. Serum follicle-stimulating hormone and luteinizing hormone concentrations that were determined in 34 individuals (24 in group 1, 10 in group 2) showed no correlation with seminal transferrin levels. In conclusion, low seminal transferrin levels correlated with low sperm density and with poor fertilizing ability of human oocytes in vitro.

Effect of vascular endothelial growth factor and interleukin-1β on apoptosis in endometrial cell cultures from patients with endometriosis and controls

The aim of this study was to evaluate the effect of vascular endothelial growth factor (VEGF) and interleukin-1beta (IL-1beta) on apoptosis induced by leuprolide acetate (LA) in endometrial epithelial cell cultures from patients with endometriosis. Primary endometrial epithelial cell cultures were obtained from uterine endometrial biopsies of patients with endometriosis and control women. Endometrial epithelial cells were incubated with LA; a combination of LA and VEGF; a combination of LA and IL-1beta; or in basal conditions. LA was added 3h prior to addition of VEGF and IL-1beta. After stimulation, the percentage of apoptotic cells was evaluated by the acridine orange-ethidium bromide technique and Bax expression was assessed by western blot. Treatment with LA enhanced the percentage of apoptotic cells in endometrial epithelial cells from subjects with endometriosis and control subjects. Addition of either VEGF or IL-1beta after exposure to LA restored the percentage of apoptotic cells to basal levels. Moreover, treatment with LA increased Bax expression in endometrial epithelial cells from patients with endometriosis. This effect was reverted by the addition of either VEGF or IL-1beta. Our results show that VEGF and IL-1beta reduce apoptosis and decrease Bax expression in endometrial epithelial cells from patients with endometriosis. This study suggests that VEGF and IL-1beta may protect endometriotic cells from undergoing apoptosis in addition to exerting their pro-angiogenic role.

Oral contraceptives treatment suppresses proliferation and enhances apoptosis of eutopic endometrial tissue from patients with endometriosis.

Fil: Meresman, Gabriela Fabiana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Instituto de Biologia y Medicina Experimental. Fundacion de Instituto de Biologia y Medicina Experimental. Instituto de Biologia y Medicina Experimental; Argentina

Influence of sequential doses of 5-hydroxytryptophan on prolactin release

It is known that the administration of serotonin or its precursors induces the release of prolactin. This study was performed (1) to determine the minimal dose of 5-hydroxytryptophan that would produce a consistent and significant prolactin increase and (2) to establish the frequency of 5-hydroxytryptophan administration necessary to induce a persistent prolactin increase. Nine normal male subjects participated in 27 independent studies following pretreatment with 100 mg of carbidopa given every 8 hours for 2 days. Doses of 0.2, 0.4, and 0.8 mg/kg/hr of 5-hydroxytryptophan were initially infused for 30 minutes, and serum prolactin was measured every 15 minutes for 2 1/2 hours. The urinary 5-hydroxyindoleacetic acid/creatinine ratio was determined in aliquots collected during 3 hours before, during, and after the intravenous infusion. 5-Hydroxytryptophan at a dosage of 0.4 mg/kg/hr was the minimal amount to elicit a consistent and significant prolactin increase (p less than 0.01). A positive correlation (r = 0.907, p less than 0.002) was also demonstrated between the maximal prolactin response and the 5-hydroxyindoleacetic acid/creatinine ratio. Thus 0.4 mg/kg/hr of 5-hydroxytryptophan was administered sequentially three times at intervals of 2, 4, 6, 8, and 12 hours. With exception of the 12-hour interval a significantly smaller plasma prolactin increase was seen following the third dose of 5-hydroxytryptophan (p less than 0.05). Furthermore, the nadir for this diminished prolactin response occurred at 4 hours (p less than 0.01). This phenomenon may represent a down regulation of the serotonin receptors induced by the repetitive administration of 5-hydroxytryptophan. In conclusion, this study has demonstrated a dose-related prolactin response to increasing doses of 5-hydroxytryptophan. The maximum down regulation of prolactin release occurred when 5-hydroxytryptophan was administered at 4-hour intervals.

Significance of ovarian macrophages in the follicular aspirates from ART patients

AbstractPurpose: To evaluate the percentages of macrophages present in granulosa cells (GC) cultures from patients with different responses to the hyperstimulation, in relation to the percentages of apoptotic cells (ApC), as well as to the release of cytokines. Methods: We studied 42 patients: 12 Hyporesponders, (with ≤4 follicles), 15 Normoresponders, (5–14 follicles), and 15 Hyperresponders, (≥15 follicles). In GC cultures percentages of macrophages and ApC were counted and, in the conditioned media, cytokines were measured Results: Percentages of macrophages were significantly higher in GC cultures from Hyporesponders compared with Hyperresponders patients. Also, the percentages of ApC cells were the highest in Hyporesponders. On the contrary, cytokines concentrations were the lowest in this group. Conclusions: The low ovarian response is probably due to the decreased angiogenesis, which in turn produces increased apoptosis and decreased production of cytokines. The increased percentage of macrophages could be related to increased frequency of apoptotic cells.