Role of galanin in the regulation of somatotrope and gonadotrope function in young ovulatory women.

The aim of the study was to elucidate the role of the neuropeptide galanin in the regulation of somatotropic and gonadotropic function in normal women. Thirteen normally ovulating (aged 28 to 40 years), non-obese (body mass index, 18.4 to 27.1 kg/m2) women with infertility due to a tubal or male factor were studied. Each woman underwent three tests: (1) bolus intravenous (IV) injection of growth hormone (GH)-releasing hormone (GHRH) (1-29)NH2 1 microgram/kg plus gonadotropin-releasing hormone (GnRH) 100 micrograms at time 0; (2) IV infusion of porcine galanin 500 micrograms in 100 mL saline from -10 minutes; and (3) bolus IV injection of GHRH(1-29)NH2 1 microgram/kg plus GnRH 100 micrograms at time 0 plus IV infusion of porcine galanin 500 micrograms in 100 mL saline from -10 to +30 minutes. All results are expressed as the mean +/- SEM. GH peak after GHRH was 14 +/- 5 micrograms/L; porcine galanin significantly increased serum GH (GH peak, 7.3 +/- 1.2) with respect to baseline levels. No significant differences were observed between either GH peak or GH absolute values after galanin as compared with GHRH alone. Porcine galanin significantly enhanced GH response to GHRH (peak, 31.4 +/- 4.4 micrograms/L) with respect to either GHRH or galanin alone. Luteinizing hormone (LH)/follicle-stimulating hormone (FSH) peaks after GnRH were 16.5 +/- 5.3 and 17.4 +/- 4 IU/L, respectively. Porcine galanin did not cause significant increases in serum LH and FSH levels with respect to baseline.(ABSTRACT TRUNCATED AT 250 WORDS)

Oocyte donation and gamete intrafallopian transfer in premature ovarian failure.

Gamete intrafallopian transfer (GIFT) was performed in eight patients with premature ovarian failure (POF), using donated oocytes. The steroid replacement protocol consisted of the administration of increasing dosages of 17 beta-estradiol (E2) and progesterone (P). Hormonal replacement was maintained until day 100 of gestation. All patients underwent an evaluation cycle in which serum levels of E2 and P were monitored and an endometrial biopsy was performed on day 21 or 22. All cases of GIFT were performed between days 12 and 15. Six clinical pregnancies were achieved in eight cycles (75% success rate). Three patients delivered and three are in their second or third trimester. No ectopics or miscarriages occurred. These results offer a promising approach for the establishment of fertility in agonadal patients.

Tubal embryo transfer as a treatment for infertility due to male factor.

Transvaginal follicular aspiration (TVA) with ultrasonically guided needles allows the transfer of in vitro generated embryos to the fallopian tubes (TET), performing only one surgical procedure in the process. Up to now, this approach has been used to treat 16 couples with infertility due to severe male factor. Follicular development was induced with a combination of clomiphene citrate and human menopausal gonadotropin (hMG) or follicle-stimulating hormone and hMG. Follicles were aspirated by TVA 36 hours after an injection of human chorionic gonadotropin 10,000 IU intramuscularly. A total of 169 oocytes were recovered (10.5 +/- 6.9 X +/- SD) from the 16 patients. There was failure of fertilization in 6 cases. In the remaining 10, a TET was performed 44 to 50 hours after TVA, utilizing embryos at the pronuclear stage. Six pregnancies resulted from the 10 transfers. This technique combines the advantages of proof of fertilization with a more adequate tubal embryo development and entrance to the uterine cavity that may determine and increase chance of implantation.

Comparison of techniques for the selection of bacteria-free sperm preparations.

The authors compared the three most commonly used sperm preparation techniques--swim-up, fall-down, and Percoll gradient--for their ability to recover highly motile sperm and minimize bacterial contamination. Eleven human semen samples collected by masturbation were used and run in parallel with the three methods. A semiquantitative bacterial analysis was performed in all samples and results expressed in colony-forming units per milliliter (CFU/ml). The Percoll gradient technique resulted in an average sperm concentration of 5.81 +/- 4.4 X 10(6) ml, and the average bacterial concentration dropped from 8.66 +/- 12.96 X 10(3) CFU/ml in semen to 0.01 +/- 0.03 X 10(3) CFU/ml. The bacterial count was not significantly different when the raw semen was compared with the swim-up or the fall-down preparations. The authors conclude that the Percoll gradient method yields an adequate sperm concentration, with high motility and improved morphology, while eliminating bacterial contamination.

[Isokinetic assessment and mid-term work reincorporation of patients surgically treated with the shoulder Latarjet technique]. / Valutazione isocinetica e reinserimento lavorativo a medio termine di pazienti operati di plastica secondo Latarjet alla spalla.

We studied a group of 12 subjects with chronic posttraumatic scapulo-humeral instability who underwent arthroscopic coracoid transposition using Latarjet's technique. All patients were followed-up after the operation for a mean of 1.7 years (minimum 1 year, maximum 3 years). The follow-ups consisted of a clinico-functional evaluation with a apprehension test and isokinetic studies of strength and joint movements during internal and external rotation of the abducted shoulder, both on the healthy side and on the operated side. The isokinetic evaluation consisted of 5 repetitions of internal-external rotation at ana angular velocity of 60 degrees/sec. The results showed that the operated shoulder was slightly weaker than the healthy one, but that the difference was not statistically significant according to Student's t test for paired data. The profile of the trace did not show macroscopic differences between the two sides. At clinical assessment 11 of 12 patients reported a good-excellent outcome of the operation with complete recovery of shoulder function and return to previously performed sporting activities. In conclusion, this technique is shown to have a good medium-term outcome. Although there have been no complications in our series, a longer follow-up to evaluate possible complications would be advisable.

CDC25A targeting by miR-483-3p decreases CCND-CDK4/6 assembly and contributes to cell cycle arrest.

Disruption of contact inhibition and serum afflux that occur after a tissue injury activate cell cycle, which then stops when confluence is reached again. Although the events involved in cell cycle entry have been widely documented, those managing cell cycle exit have remained so far ill defined. We have identified that the final stage of wound closure is preceded in keratinocytes by a strong accumulation of miR-483-3p, which acts as a mandatory signal triggering cell cycle arrest when confluence is reached. Blocking miR-483-3p accumulation strongly delays cell cycle exit, maintains cells into a proliferative state and retards their differentiation program. Using two models of cell cycle synchronization (i.e. mechanical injury and serum addition), we show that an ectopic upregulation of miR-483-3p blocks cell cycle progression in early G1 phase. This arrest results from a direct targeting of the CDC25A phosphatase by miR-483-3p, which can be impeded using an anti-miRNA against miR-483-3p or a protector that blocks the complex formation between miR-483-3p and the 3'-untranslated region (UTR) of CDC25A transcript. We show that the miRNA-induced silencing of CDC25A increases the tyrosine phosphorylation status of CDK4/6 cyclin-dependent kinases which, in turn, abolishes CDK4/6 capacity to associate with D-type cyclins. This prevents CDK4/6 kinases' activation, impairs downstream events such as cyclin E stimulation and sequesters cells in early G1. We propose this new regulatory process of cyclin-CDK association as a general mechanism coupling miRNA-mediated CDC25A invalidation to CDK post-transcriptional modifications and cell cycle control.

Validation of a novel approach for the rapid production of immunogenic virus-like particles for bluetongue virus.

Bluetongue virus causes an emerging disease of ruminants, principally affecting sheep and cattle. Since 1998, there have been multiple separate outbreaks of bluetongue disease in Europe that have highlighted the need for a safe, efficacious, DIVA compliant vaccine. We report here a new baculovirus expression strategy which allowed pre-integration of the genes encoding the BTV inner capsid proteins at one baculovirus locus and those encoding the outer capsid proteins at a different locus. A modified baculovirus with two marker proteins to facilitate the phenotypic selection of recombinant viruses was developed. The utility of this approach is demonstrated by the production of BTV VLPs to a number of serotypes. For a proof of concept, VLPs of one serotype was then tested for protective immune response. VLPs were demonstrated to be safe, highly effective, immunogens in sheep, reducing post-challenge viraemia to levels below the threshold detection limit of quantitative RT-PCR when vaccinated animals were challenged with virulent virus.

A new association of ethinylestradiol (0.035 mg) cyproterone acetate (2 mg) in the therapy of polycystic ovary syndrome.

The authors treated 28 patients with polycystic ovary syndrome (PCOS) with the new preparation SH B 209 AE consisting of 0.035 mg of ethinylestradiol and 2 mg of cyproterone acetate (EE + CPA) throughout 12 months. From the analysis of the endocrine and clinical modifications induced by the drug along with the negligible incidence of side effects the conclusion can be drawn that this new estro-progestational association is indicated in the therapy of PCOS.

PIP: 28 patients with polycystic ovary syndrome were treated for 12 months with the new preparation SH B 209 AE, consisting of 0.035 mg of ethinyl estradiol and 2 mg of cyproterone acetate. This was the first clinical trial of estroprogestational therapy on a homogeneous sample of women with polycystic ovary syndrome. Endocrine findings indicated a significant decrease in all hormonal parameters, the invariableness of prolactinemia, a considerable increase in sex hormone binding globulin (SHBG) at the 6th treatment cycle examination, a continuous significant decrease in 17 beta E2 and androstenedione from the 6th to the 12th treatment cycles. In terms of clinical findings, there was a significant decline in the severity of acne, seborrhea, and hirsutism during drug administration. The menstrual cycle in the 28 study subjects remained under control during treatment, and there were no pregnancies. Side effects such as weight gain, nausea, headache, and changes in libido were not reported. Overall, the findings of this study suggest that administration of the new monophasic contraceptive association SH B 209 AE can normalize endocrine patterns in polycystic ovary syndrome and improve its androgenic symptomatology. The low content of estrogen, the changes in clinical and hormonal parameters, the low incidence of side effects, and the good control of the menstrual cycle provided by this treatment make SH B 209 AE deserving of more widespread application.

A new method for free testosterone assay: clinical evaluation.

The Authors propose a new direct method for the determination of free Testosterone (F.Te). Our study was made following this method: 1) on a control group composed of 12 healthy men and 21 healthy women, and 2) on a group of 29 patients suffering from Polycystic Ovary Syndrome (PCOS) with clinical signs of hyperandrogenism. This pathological group presented acne and hirsutism in 95% of the cases. The Authors demonstrate how the determination of F.Te permits a 93.1% correct endocrinological diagnosis of hyperandrogenism.

[A study of biochemical parameters in 94 athlets of Marcialonga (author's transl)]. / Studio di alcuni parametri biochimici in atleti della Marcialonga.

Twenty biochemical parameters have been studied in 94 athlets of Marcialonga in basic conditions and after 30 min from the end of the competition. Urea, uric acid, creatinin, total proteins, albumin, sodium, calcium, phosphorus and several enzymes (AST, ALT, LAD, CK and ALP) have shown statistically significant increasings. It is, above all, clear the increasing of CK. On the contrary triglycerides have undergone a significant decreasing. The AA. try to explain the results obtained, considering some pathogenetic theories.

[Incidence of salmonellosis in the Province of Trento in the past 3 years]. / Incidenza delle salmonellosi in provincia di Trento nell'ultimo triennio.

72 strains of Salmonella, belonging to 23 different serum types have been isolated during the last three years. These strains come from adult patients admitted to the Infective department of St. Chiara hospital in Trento and from three asymptomatic carriers found among the hospital staff, who have been regularly submitted to microbiological checks. Among the most frequent strains the AA. have found: S. enteritidis and S. typhi murium. Salmonellas, never isolated before, have spread, but they have shown poor adaptation to the environment: among these S. panama, S. infantis, S infantis, S. münchen. S. wien has not found, in the Province of Trento, environing conditions fit for its colonization. S. agona has shown a certain tendency to create a situation of asymptomatic carrier.

Tubal embryo transfer in cynomolgus monkeys: effects of hyperstimulation and synchrony.

To date the main limitation of in-vitro fertilization-embryo transfer (IVF-ET) programmes is that transcervical transfer of embryos results in a rate of low implantation. On the other hand, the technique of gamete intra-Fallopian transfer (GIFT) does not contribute to information on oocyte fertilization rates, and the time of oocyte exposure to sperm may be limited. The development of ultrasonically guided follicular aspiration will allow transfer of embryos generated in vitro to the Fallopian tubes performing only one surgical procedure in the process. We have performed 25 intra-tubal embryo transfers in the cynomolgus monkey (Macaca fascicularis). Ovarian stimulation, follicular aspiration and IVF procedures have been reported previously by Balmaceda. ETs were performed via laparotomy. Embryos at the 2-, 4- or 9-cell stage were loaded into a tom-cat catheter in 5 microliter of culture medium and delivered to the mid-ampullary portion of the tube. Seven ETs performed during stimulated cycles resulted in one pregnancy, and 18 ETs performed in synchronized recipients resulted in six pregnancies. Ten ETs were performed 0-24 h, eight performed 24-48 h and seven performed 72-110 h after follicular aspiration or ovulation, and resulted in 4, 3 and 0 pregnancies respectively. Our results demonstrate that intra-tubal embryo transfer can result in normal intra-uterine pregnancies and suggest that both ovarian stimulation and cycle synchronization affect the probability of embryo implantation.

Comparison of a single-antigen microimmunofluorescence assay and inclusion fluorescent-antibody assay for detecting chlamydial antibodies and correlation of the results with neutralizing ability.

An inclusion fluorescent-antibody assay (IFA) with McCoy cells infected with Chlamydia trachomatis serovar L2 was compared with a single-antigen (L2) microimmunofluorescence (MIF) assay for the detection of antichalmydial antibodies. A total of 562 serum specimens were tested by both assays, and sera representing a range of titers were tested for their ability to neutralize the infectivity of C. trachomatis. Overall, there was poor correlation between the two assays (r2 = 0.62). With most sera the inclusion IFA was more sensitive. There was better correlation between IFA titer and ability to neutralize the five serovars tested (L2, L3, C, E, and F) than between the MIF assay and neutralization. In summary, the IFA appeared to be more sensitive than the MIF assay for detecting antibodies to C. trachomatis.

Intrauterine insemination and controlled ovarian hyperstimulation in cycles before GIFT.

We have combined intrauterine insemination (IUI) and controlled ovarian hyperstimulation (COH), for the treatment of infertility due to different aetiologies, prior to performing GIFT. To date, we have treated 186 patients over a total of 489 cycles. The mean age of the patients was 34.1 +/- 4 years and the mean duration of infertility was 4.8 +/- 2.8 years. Follicular development was induced with human menopausal gonatrophin (HMG). Patients were monitored using serum oestradiol determinations and ovarian ultrasound. Two intrauterine inseminations were performed 12 and 36 h after HCG injection. Semen samples were prepared utilizing one of two techniques, swim-up or Percoll gradient. A total of 33 pregnancies have occurred, the gross pregnancy rate being 17.7% per patient and 6.7% per cycle. The cumulative pregnancy rate was 30%. Thirty-one pregnancies (94%) occurred within the first four cycles of treatment. During the same period of time, the pregnancy rate per cycle in patients treated with gamete intra-Fallopian transfer (GIFT) was 32.9%. Our data suggest that IUI combined with COH can result in pregnancy in a significant proportion of patients, but that the efficiency per cycle of the technique is significantly lower than GIFT.