Carlo Flamigni

The successful use of human amniotic fluid for mouse embryo culture and human in vitro fertilization, embryo culture, and transfer

The development of mouse and human embryos was assessed in human amniotic fluid to determine its suitability as a culture medium for human in vitro fertilization (IVF). Two-cell mouse embryos developed to blastocysts after 72 hours at rates similar to that in Whittinghams T6 + 10% fetal calf serum. Significantly more mouse embryos hatched in amniotic fluid. No difference was found between individual patients amniotic fluids obtained at 16 to 21 weeks gestation. A preliminary trial comparing amniotic fluid with T6 + maternal serum in human IVF showed no significant difference in fertilization rate and embryo development during 42 to 48 hours in vitro. Expanded blastocysts were obtained in amniotic fluid after 5 days in vitro. Four pregnancies were obtained in 9 patients transferred embryos grown in amniotic fluid and with 2 or 12 patients transferred embryos grown in T6 + maternal serum.

Ongoing Pregnancy After Intracytoplasmic Sperm Injection of Epididymal Spermatozoa into Cryopreserved Human Oocytes

Human embryo cryopreservation presents several ethical, legal, and religious problems and is a matter of discussion in several countries. The possibility of storing human oocytes would virtually eliminate these problems and, in addition, could give a reproductive chance to young patients at risk of losing ovarian function. However, only sporadic clinical success has been reported in the past decade (1). The recent technical improvement (2) and clinical success (3) have renewed interest in oocyte cryopreservation, whose possible applications are expanding. The present report describes the first pregnancy achieved with intracytoplasmic injection of epididymal sperm into frozenthawed eggs.

T‐Helper 2 Type Cytokine and Soluble Interleukin‐2 Receptor Levels in Seminal Plasma of Infertile Men

PROBLEM: The role of cell‐mediated immunity (CMI) in unexplained male infertility and impaired sperm function has been explored.

A histological and immunohistochemical study of neuropeptide containing somatic nerves in the levator animuscle of women with genitourinary prolapse

BACKGROUNDnTo evaluate the changes occurring in the somatic innervation throughout the levator ani muscle in women with genitourinary prolapse and stress urinary incontinence (SUI).nnnMETHODSnThirty-four patients with genital prolapse entered the study and ten subjects with non-malignant pathologies acted as a control group. All patients were evaluated by urodynamic investigation and an electromyographic study of pelvic floor muscles to define the type of urinary incontinence. Biopsy samples were obtained from both groups of patients: the site of muscle biopsies were left and right part of perirectal pubococcygeus muscle. The evaluation of immunoreactivity was semiquantitative and based on staining intensity and distribution.nnnRESULTSnIn all cases, S-100 protein and NSE immunoreactivities were found in nerve fascicles running throughout the striated muscle. NPY and VIP positivities were more intense and diffuse, whereas SP immunoreactivity was quite scanty. The different patterns of NPY and VIP expression changed in relation to degree of genital prolapse and to the presence of SUI.nnnCONCLUSIONSnOur immunohistochemical study shows the presence in the pelvic floor of neurons that are able to synthesize neuropeptides. The lower immunoreactivity score of same neuropeptides (VIP, NPY) observed among patients with third degree genital prolapse and with SUI could be related to biochemical damage of the neurons with subsequent lower production of chemical messengers.

Characterization of human sperm antigens reacting with antisperm antibodies from autologous sera and seminal plasma in a fertile population

Immunoblotting techniques were used to characterize the reactivity of human sperm antigens with antisperm antibodies from a population of fertile individuals. In particular, sperm antigens of each subject were tested with the same subjects antisperm antibodies present in blood serum and seminal plasma in an attempt to construct a preliminary map of the antigen domains of the normal spermatozoon. Fifty-five fertile males, comprising 22 subjects with a pregnant partner and 33 subjects attending assisted reproductive technology sessions for proven partners infertility and with normal semen quality entered the study. A high proportion of sera (82%) and seminal plasma (62%) showed antisperm antibodies reacting with one or more sperm antigens. Specific immunoreactivity was often demonstrated to 45-kDa, 50-kDa, 55-kDa, 69-kDa, 72-kDa and 85-kDa proteins in serum and to 59-kDa and 72-kDa proteins in seminal plasma. These proteins are the most frequently involved sperm antigens in the immune responses in fertile subjects. Further studies in an infertile population are necessary to distinguish between these antigens of minor relevance in sperm function from others significantly involved in immunological infertility.

Polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a complex and still poorly understood process. This review examines current theories regarding the development of PCOS and focuses on the physiologic processes involved in the pubertal and adolescent period and their relationship with pathogenesis of PCOS.

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.

Endometriosis: Absence of Recurrence in Patients After Endometrial Ablation

Endometriosis, a very common and debilitating illness, has responded to both medical and surgical treatments, but optimal treatment remains unavailable. This study was planned to determine whether patients with and thosewithout eutopic endometrium differ in the risk that ectopic endometriotic implants will recur. Of 28 women 24 to 40 years of age with at least one child who had a surgical diagnosis of endometriosis after uncontrolled dysmenorrhea, 14 underwent hysteroscopic endometrial ablation (EA). For the procedure, a roller ball was used at 50 watts in the hope of avoiding recurrent abdominal dissemination of endometrial debris. The remaining patients had laparoscopic treatment of endometriosis only, using either bipolar instruments or excision of the implants. Laparoscopy was repeated 2 years later. Nine of 14 patients undergoing EA had no dysmenorrhea 3 months later, and 3 others had significantly less bleeding. Symptoms were appreciably lessened in only 3 of the 14 non-EA group. No retrograde bleeding was seen in EA patients at follow-up laparoscopy, but 9 of the 14 not having EA had significant recurrences. Stromal and epithelial endometrial cells were found within debris in the cul-de-sac in eight patients not having EA. These results support a critical role for removing eutopic endometrium to prevent recurrent endometriosis. The therapeutic key may be to stop the retrograde uterine contractility during menstruation that may be the major cause of recurrence through ectopic endometrial dissemination. Recurrences may result from treatments that are mainly directed to deleting endometriotic implants and to reducing or antagonizing promoters.

Andrology: 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.

Evidence for Pathological Reduction in Brain Dopamine Metabolism in Idiopathic Hyperprolactinemia

The role of brain catecholamine activity in the neuroendocrine regulation of the dopamine-PRL system in idiopathic hyperprolactinemia was investigated by high-performance liquid chromatography with electrochemical detector. We measured urinary dopamine, norepinephrine, epinephrine, vanillylmandelic acid, homovanillic acid, 3,4-dihydroxyphenylacetic acid and total 3-methoxy-4-hydroxyphenylglycol levels in 12 women with idiopathic hyperprolactinemia before and during either peripheral dopa-decarboxylase blockade, by carbidopa, or dopamine beta-hydroxylase blockade, by disulfiram. Homovanillic acid and 3,4-dihydroxyphenylacetic acid concentrations were significantly lower (p less than 0.001 and p less than 0.005, respectively) in patients with idiopathic hyperprolactinemia compared with those in 12 control subjects in the early follicular phase, whereas they were similar to those in the control subjects in the pre-ovulatory phase. Dopamine, norepinephrine, epinephrine, vanillylmandelic acid and 3-methoxy-4-hydroxyphenylglycol concentrations were similar to those of the control subjects in both phases of the cycle. During carbidopa administration the levels of all urinary catecholamines and metabolites were unchanged, except that of dopamine which dropped remarkably (p less than 0.001). During disulfiram administration dopamine, homovanillic acid and 3,4-dihydroxyphenylacetic acid concentrations increased (p less than 0.05, p less than 0.001 and p less than 0.005, respectively) and those of norepinephrine, vanillylmandelic acid and 3-methoxy-4-hydroxyphenylglycol decreased (p less than 0.05, p less than 0.001 and p less than 0.005, respectively), whereas epinephrine levels remained unaltered. These data support the existence of a quantitatively reduced brain dopamine activity in idiopathic hyperprolactinemia.