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Biomedical subjects

C Flamigni

Publications and source records attributed to C Flamigni.

At least 91 records · Page 5Linked to original sources

Dating the endometrial biopsy by flow cytometry.

OBJECTIVE: To determine endometrial changes throughout the menstrual cycle. DESIGN: Flow cytometric analysis of endometrial cells versus chronological dating. SETTING: Women volunteers with a normal menstrual cycle participated in this study that was done in an academic research environment. PATIENTS: Two hundred thirty regular menstruating women with adequate luteal phase underwent endometrial biopsy at different days of their menstrual cycle; 138 biopsies were analyzed both histologically and by flow cytometry. MAIN OUTCOME MEASURE: Percentage of biopsies correctly classified in terms of chronological dating. RESULTS: Flow cytometry correctly classified the chronological day of biopsies in 59% of cases, the proliferative versus secretory days in 91% of cases, and the early, mid, and late secretory phases of the menstrual cycle were correctly predicted in 86% of cases. CONCLUSION: Flow cytometry is a good method to determine growth and differentiation of the endometrium during the menstrual cycle. Detection of phases of the whole endometrial cell population offer information on the biological adequacy of endometrial response to the biochemical environment.

Biopsy↗

The role of gamete intrafallopian transfer (GIFT) and tubal embryo transfer (TET) in the treatment of patients with patent tubes associated with male infertility factor.

PURPOSE: The relative effectiveness of gamete intrafallopian transfer (GIFT) and tubal embryo transfer (TET) combined with superovulation in the treatment of infertile patients with patent tubes were compared. Four hundred fifty consecutive cycles were divided into two periods. During the first period (216 cycles), the only technique employed was GIFT, couples being divided into two groups: group A, couples with normospermic partners (118 cycles); and group B, couples with male infertility factor (98 cycles). During the second period (234 cycles), 140 cycles of GIFT were performed in couples with normospermic partners (group C). TET was utilized in 94 cycles (group D), in the case of couples with male infertility factor. RESULTS: Results demonstrate that the pregnancy rate with GIFT in the case of oligoasthenospermic partners (group B) is significantly lower than that of normospermic partners (groups A and C) (P = 0.0001) and than that with TET in the case of oligoasthenospermic partners (group D) (P = 0.0001). CONCLUSION: The implantation rate is also significantly different between these groups (B vs A, P = 0.0001; B vs C, P = 0.0001; B vs D, P = 0.01).

Embryo Transfer↗

Immunoreactive dynorphin A-like material in extracted human hypothalamic-hypophysial plasma.

Immunoreactive dynorphin A-like material (ir-dyn A) in human plasma was measured by a validated radioimmunoassay. In peripheral plasma extracts mean concentrations between 20 and 40 fmol/ml were determined in volunteers and in patients with pituitary adenomas. In this latter group superimposable levels were detected three days before and during transsphenoidal microsurgery. Interestingly, ir-dyn A levels evaluated in extracts of hypothalamic-hypophysial plasma obtained during surgery, just after tumor removal, were 4-5 times higher than in peripheral plasma. Reverse-phase high performance liquid chromatography (rp-HPLC) of extracts of peripheral plasma samples revealed two immunoreactive peaks. The major form had the same retention time of dyn A-(1-32); whereas a second, more lipophilic, peak eluted later and was not further characterized. In contrast, rp-HPLC analysis of extracts of plasma collected from the suprapituitary region displayed only one peak eluting in the position of synthetic dyn A-(1-17). The presence of dyn-related peptides in hypothalamic-hypophysial plasma supports the hypothesis that they may play a part in the regulation of hypothalamic and/or pituitary functions in humans.

Adenoma↗

Electromechanical activities of human uteri during extra-corporeal perfusion with ovarian steroids.

A new experimental system was designed to study human uterine activities based on the extra-corporeal perfusion of isolated human uteri. Electromechanical activities in the uterine wall were recorded using bipolar silver-silver electrodes, endoluminal pressure catheters and a dedicated acquisition, storage and analytical system. The electrical signals recorded were isolated spikes and rhythmic activities; the last being primarily associated with organized mechanical events. Perfusion media containing 17 beta-oestradiol alone or with progesterone were used for those uteri obtained during proliferative (n = 5) or secretory (n = 5) phases of the menstrual cycle, respectively. Progesterone caused a reduction of frequency (P < 0.001) and duration (P < 0.001) of the rhythmic electrical activity, and decreased the endoluminal pressure at both detection sites (P < 0.01). 17 beta-Oestradiol increased both frequency (P < 0.001) and duration (P < 0.001) of the rhythmic electrical activity as well as the endoluminal pressure at two different detection sites (3 and 5 cm from the fundus) (P < 0.05). Significant differences between the fundus and cervix sites in the uterine wall were detected. In conclusion, uterine perfusion would be useful to examine the effects of uterotonic and tocolytic drugs before administration to humans, at no risk to the patients. Oestrogens increase and progesterone decreases both electrical and mechanical uterine activities.

Action Potentials↗

Oocyte donation: comparison between recipients from different age groups.

Oocyte donation was carried out in 87 patients in 141 replacement cycles. These patients received oocytes from 108 women undergoing assisted reproductive technology procedures at our centre. Standardized hormonal replacement therapy and in-vitro fertilization procedures were performed. We divided recipients into four groups according to their age (group A, 21-35 years; B, 36-40 years; C, 41-49 years; and D, 50-61 years). Oocytes donors were 21-35 years old, and equally spread across these different age groups. There were significant differences in the pregnancy and implantation rates according to the age of the recipients; which were 45% and 23% respectively in women 21-35 years old (group A) versus 23% and 10% in women 41-49 years old (group C). A comparison of data between oocyte donors and young recipients, with similar results in donors and young recipients, with pregnancy rates of 45% and 42% and implantation rates of 23% and 19.5% respectively. Statistically significant differences were found between donors and the older recipients, pregnancy rates being 43% versus 23%, and implantation rates 18% versus 10%. These data seem to demonstrate a lesser likelihood of pregnancy and implantation in older recipients because of increasing uterine age.

Adult↗

A new approach to gamete intra-fallopian transfer via hysteroscopy.

This study describes the technique and results of a new approach for the hysteroscopic catheterization of the Fallopian tube in order to transfer gametes (hysteroscopic GIFT). Ovarian stimulation was started on 51 patients. Three cycles were cancelled because of a failure of ovarian response. Forty-eight patients underwent hysteroscopic GIFT utilizing a technique completely different, easier, faster and more precise than that previously reported. Fourteen pregnancies were achieved (29.2%) of which five are ongoing, six patients have delivered and three have aborted. No ectopic pregnancies were observed.

Adult↗

Influence of spermatozoa characteristics on gamete intra-fallopian transfer procedures: analysis of results obtained utilizing normozoospermic, oligoasthenozoospermic and donor spermatozoa.

The objective of this study was to analyze the efficacy of the gamete intra-Fallopian transfer (GIFT) procedure in relation to spermatozoa characteristics. A total of 268 infertile couples enlisted for GIFT were categorized into three groups on the basis of semen characteristics in the husband. These included oligoasthenozoospermic men in 88 retrieval cycles (group 1), normozoospermic men in 116 cycles (group 2) and azoospermic partners who needed donor semen in 86 cycles (group 3). All female patients had ovarian stimulation and laparoscopic GIFT. Pregnancy rates were significantly higher with donor spermatozoa than with oligoasthenozoospermic or normozoospermic spermatozoa, i.e. 51.1% versus 15.9% (P < 0.001), and 32.7% (P < 0.005). Implantation rates were significantly higher with donor spermatozoa than with normo- or oligoasthenozoospermic spermatozoa (P < 0.01). These data suggest that GIFT does not give good results with male factor infertility. Donor cryopreserved semen gives higher pregnancy and implantation rates than normozoospermic semen, and GIFT with donor spermatozoa gives a good chance of pregnancy to couples previously treated with artificial insemination using donor semen.

Female↗

Changes in brain catecholamine metabolism during bromocriptine treatment in polycystic ovary syndrome.

The role of dopaminomimetic drugs on the brain catecholamine metabolism in the neuroendocrine regulation of the polycystic ovary syndrome (PCO) was investigated. We measured, besides peptide hormones and sex steroids, urinary dopamine (DA), norepinephrine, epinephrine, vanillylmandelic acid, homovanillic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC) and total 3-methoxy-4-hydroxyphenylglycol (MHPG) levels by high-performance liquid chromatography with electrochemical detector in 10 women with PCO before and during long-term bromocriptine (BRC) administration. HVA and DOPAC concentrations were significantly lower (p < 0.001) in PCO patients compared with 12 control subjects in the early follicular phase, whereas MHPG concentrations were significantly higher (p < 0.01) in PCO patients. During BRC administration, HVA, DOPAC and MHPG levels increased significantly (p < 0.01 for HVA and DOPAC, and p < 0.05) for MHPG), prolactin levels dropped markedly (p < 0.01), whereas luteinizing hormone levels did not change (p = NS). These data show (1) a reduced DA activity in PCO which may be normalizable under BRC treatment, but also (2) no major effects of DA metabolism on the inappropriate gonadotropin secretion of the syndrome.

3,4-Dihydroxyphenylacetic Acid↗

Comparison of the suppressive capacity of different depot gonadotropin-releasing hormone analogs in women.

Different depot GnRH analogs (GnRH-A) are currently used for the reversible suppression of the pituitary-ovarian axis in several reproductive and neoplastic disorders in women. In spite of anecdotal reports of incomplete suppression by some depot GnRH-A, this issue has never been systematically investigated in adult women. Thus, we elected to study 40 normally cycling women with male-related infertility or benign reproductive disorders; each group of 10 subjects received a different GnRH-A for 3 months: buserelin (group B; 300 micrograms, sc, every 12 h, as a control), goserelin (group G; 3.6 mg, sc, every 28 days), leuprorelin (group L; 3.75 mg, im, every 28 days), and triptorelin (group T; 3.75 mg, im, every 28 days). Depot GnRH-A was administered by one of the investigators. GnRH tests (100 micrograms, iv) were performed before treatment (cycle day 7; test A) and on treatment days 57 (i.e. 1 day after the third depot GnRH-A; test B) and 84 (i.e. 28 days after the third depot GnRH-A; test C). Immunoreactive (i) LH levels were measured with an ultrasensitive immunochemiluminometric assay. Profound suppression of the iLH response to the GnRH test occurred in all subjects during treatment. Conversely, FSH levels in the third month of treatment tended to be higher in the depot GnRH-A groups than in group B, and this difference achieved statistical significance (P < 0.05) in groups G and L during test C. In GnRH test B, while the mean estradiol (E2) level was less than 75 pmol/L (< 20 pg/mL) in all group B subjects, individual E2 levels were greater than 75 pmol/L in five patients receiving depot GnRH-A (two in group G, one in L, and two in T). Finally, individual E2 levels during test C were greater than 75 pmol/L in only two patients of group G, who also reported vaginal spotting. Thus, we conclude that in adult women, 1) iLH was profoundly suppressed in the third month of administration of all GnRH-A tested; 2) FSH suppression with depot GnRH-A was less marked than that with high-dose short-acting sc buserelin; and 3) signs of an incomplete block of ovarian function can be present in the third month of depot GnRH-A administration, particularly when goserelin is employed.

Adult↗

Adeno-pituitary hormones in human hypothalamic hypophysial blood.

An in vivo technique for collecting blood from the pituitary stalk using transphenoidal microsurgery has recently been developed in men with nonfunctioning pituitary disease. To determine the origin of this blood and the direction of the stream, we measured contemporaneously the levels of LH, FSH, PRL, GH, TSH, and ACTH in hypothalamic-hypophysial blood (HHB) and peripheral blood (PB). Eleven patients with nonfunctioning pituitary adenomas entered the study. The surgical procedure used for collecting HHB consisted of periodically aspirating small amounts of blood using a microsuction apparatus, just after tumor removal, kept in the postero-superior corner of the sella turcica at the junction of the diaphragm with the dursum sellae. The data show clearly the existence of a dramatic concentration gap in HHB vs. PB in all adeno-pituitary hormones (P = 0.003). The HHB/PB ratio varied from 50-600 in the different hormones. The secretion of adeno-pituitary hormones in blood drawn at the pituitary stalk level in man was reported for the first time. The dramatic HHB/PB ratio of the hormone levels has been emphasized. The most likely explanation for the markedly elevated hormone concentration gradient between central and peripheral blood was sampling of peri- and/or suprapituitary blood. To consider the origin and direction of the HHB stream, two hypotheses have been further advanced: 1) a retrograde bloodflow from the pituitary, and 2) a central-hypothalamic secretion.

Adenoma↗

Longitudinal evaluation of the different gonadotropin pulsatile patterns in anovulatory cycles of young girls.

We studied 13 adolescents (mean gynecological age 29.2 +/- 14.1 months) with anovulatory cycles and 7 women with ovulatory cycles (mean gynecological age 33.1 +/- 15.3 months) as a control group. Adolescents with anovulatory cycles were grouped on the basis of mean plasma LH values: group 1 (n = 7) with high LH values, and group 2 (n = 6) with normal LH values. In all women plasma gonadotropin concentrations were measured at 10-min intervals for 8 h on day 4 of the cycle. Pulsatile gonadotropin secretion was also studied in each subject a second time 40 months later, to establish the outcome of the different pulsatile patterns. Group 1 had more frequent and greater LH pulses than the other two groups (which were similar) and had the highest plasma 17 beta estradiol, testosterone, androstenedione, and 17 hydroxyprogesterone concentrations. Longitudinal control showed that: in group 1, three subjects out of seven acquired ovulatory cycles and there was a fall in mean LH plasma levels (30 +/- 5 vs. 9 +/- 4 IU/L; P less than 0.01), number of pulses (8.3 +/- 1.5 vs. 5 +/- 0; P less than 0.025), mean amplitude (13 +/- 3 vs. 5 +/- 2 IU/L; P less than 0.02) and an increase in interpulse interval (56 +/- 10 vs. 91 +/- 6 min; P less than 0.01). In four subjects anovulatory cycles persisted and the LH pulsatile profile remained unchanged. In group 2, five subjects out of six acquired ovulatory cycles, but there were no significant changes in the number of pulses (6 +/- 1 vs. 6 +/- 2; P = NS), interpulse interval (97 +/- 30 vs. 85 +/- 30 min; P = NS), or amplitude (5 +/- 2 vs. 4 +/- 2 IU/L; P = NS). The results indicate that: 1) anovulatory young women with early normal plasma LH values have an adequate GnRh pulsatile pattern which will easily lead to ovulation; 2) anovulatory young women with high LH plasma values may have a reproductive system blocked in a pathological condition, similar to that observed in polycystic ovary syndrome; 3) only few subjects with high plasma LH values are able to achieve ovulation and normalize LH pulsatile pattern as a consequence of a new mode of GnRh release.

Adolescent↗

Gamete intrafallopian transfer by hysteroscopy as an alternative treatment for infertility.

OBJECTIVE: To evaluate efficacy and safety of the hysteroscopic cannulation by flexible catheter of the fallopian tubes for gamete intrafallopian transfer (GIFT). DESIGN: We studied the pregnancy rate (PR) and the safety of this new technique. SETTING: All patients were enlisted for GIFT at our Reproductive Medicine Unit. PATIENTS: We treated 26 patients whose infertility causes were terminal tubal damage, male factors, unexplained factors, and endometriosis. Patients with uterine tubal ostia unsuitable for gamete transfer or cervical incontinence were not included in the group. INTERVENTIONS: The patients underwent ovulation induction and oocyte retrieval by transvaginal ultrasonically guided puncture. The gamete transfers were carried out by hysteroscopic procedure using a flexible catheter put through the operating channel. MAIN OUTCOME MEASURE: The efficacy was evaluated by the PR (25.9%). RESULTS: Seven clinical pregnancies were obtained, but two patients aborted during the first weeks of pregnancy. No ectopic pregnancies were observed. CONCLUSIONS: Our results indicate that hysteroscopic GIFT is an alternative, safe, effective, and not invasive technique for fertility problems.

Adult↗

Hysteroscopic gamete intra-fallopian transfer: a good alternative, in selected cases, to laparoscopic intra-fallopian transfer.

Results obtained in two groups of patients treated with gamete intra-Fallopian transfer (GIFT) are reported. Hysteroscopic GIFT was carried out in some cases where general anaesthesia was not advisable or possible, i.e. difficulties in tubal catheterization due to pelvic adhesions, extended distal tubal damage, patients' intolerance, lack of available operating theatre. Ovarian stimulation was started on 131 patients for a total of 147 cycles. Twenty-five cycles were cancelled because of failure of the ovarian response. Either laparoscopic (group 1; 73 patients) or hysteroscopic GIFT (group 2; 50 patients) was performed. In group 1 a mean of 6.8 +/- 3.4 oocytes per cycle were retrieved and a mean of 4.7 +/- 1.3 mature oocytes were transferred. The pregnancy rate was 30.1% per retrieval (22 clinical pregnancies). In group 2, a mean of 5.0 +/- 3.1 oocytes was harvested and 3.9 +/- 2.0 mature oocytes per cycle were transferred. Thirteen clinical pregnancies were achieved (26.5% per cycle) in group 2. Comparing the two groups, the number of recovered and transferred oocytes was significantly lower in hysteroscopic GIFT (P less than 0.05 and P less than 0.001 respectively).

Adult↗

Pregnancy after direct intraperitoneal insemination.

Direct intraperitoneal insemination (DIPI) and superovulation are simple procedures which may together represent a good alternative to gamete intra-Fallopian transfer (GIFT) in infertile women with patent Fallopian tubes. In the present study, pregnancy occurred in 25 of 96 couples (26%) and six (24%) of these aborted. The pregnancy rate for all cycles was 19.6% and multiple pregnancies were found in six of 25 (24%) patients. We observed no ectopic pregnancy. The combination of these techniques is concluded to be useful in achieving pregnancy in infertile women with patient Fallopian tubes.

Adult↗

Factors regulating interaction between trophoblast and human endometrium.

Implantation is a crucial step in human reproduction. Disturbances of this process are responsible for pregnancy failure after both in vivo and in vitro fertilization. The endometrium provides the implanting embryo with a unique substratum where the embryo communicates with biochemical signals, attaches itself, penetrates and grows without blood circulation. The highly proliferative phase of the cytotrophoblast, during early human embryogenesis, may be due to endogenous production of growth factors that may establish autocrine/short range paracrine stimulator loops which explain the tumor-like properties of these tissues. Endometrial BM penetration and stroma invasion may be due to the proteolytic capability of the human embryo. It is suggested that collagenase and the urokinase-like plasminogen activator are responsible for this activity. To clarify the molecular mechanisms involved in human embryo implantation several models are suggested: culture of blastocysts, culture of endometrial cells, and endometrial explant co-culture. Human blastocysts cultured with whole perfused human uteri make it possible to recognize some aspects of the entire implantation process and give us the possibility of improving the benefits provided by new technologies in reproductive medicine and reducing embryonic loss at an early stage.

Endometrium↗

Tissue factors influencing growth and maintenance of endometriosis.

The unpredictable response of endometriosis to steroids and its recurrence after therapy, led us to hypothesize a possible further control of this pathology by factors other than steroids. The presence of estrogen, progesterone and epidermal growth factor receptors (ER, PR, EGFr) was evaluated using immunohistochemistry before and after therapy with Danazol or a gonadotropin-releasing hormone analogue (GnRHa), Buserelin. EGFr, ER and PR were present in 100% of endometrial specimens, and in 71%, 29% and 49% of endometriotic implants, respectively. Danazol and GnRHa reduced immunohistochemical staining for EGFr antisera in the endometrial and endometriotic specimens. About 21% of endometriosis were EGFr positive and ER negative, suggesting a potential role of epidermal growth factor in growth and maintenance of endometrial ectopia.

Adult↗