Oocyte vitrification modifies nucleolar remodeling and zygote kinetics-a sibling study.

PURPOSE: Oocyte vitrification does not affect embryo quality after oocyte warming, making this method effective in the preservation of female fertility. Morphokinetic parameters can be used to predict the competence of an embryo produced from fresh oocytes. Our aim was to study the effect of oocyte vitrification on zygote-embryo kinetics (pl). METHODS: The embryo-kinetics of fresh and sibling vitrified/warmed oocytes were compared to determine the consequences of oocyte preservation on the timing of embryo development. A 44-hours time-lapse analysis, from the time of ICSI (t0), of 179 fertilized fresh oocytes was compared to 168 fertilized sibling vitrified/warmed oocytes. RESULTS: Oocyte vitrification accelerated pronuclear disappearance, one-cell stage timing and modified nucleoli activity by increasing their number and decreasing their diameter at the zygote stage. In contrast, embryo kinetics during cleavage were similar to those observed for fresh sibling oocytes based on the parameters examined in this study. CONCLUSIONS: At the zygote stage, oocyte vitrification induces changes in pronuclei stability, probably due to pronuclei envelop instability as well as modifications in nucleoli functionality. Therefore, the predictive morphokinetic parameters on embryo competence found from fresh oocytes must be revised when applied on embryos from vitrified/warmed oocytes.

Effect of gestational hypercholesterolaemia on omental vasoreactivity, placental enzyme activity and transplacental passage of normal and oxidised fatty acids.

OBJECTIVE: Maternal hypercholesterolaemia during pregnancy increases lipid peroxidation in mothers and fetuses and programs increased susceptibility to atherosclerosis later in life. The objective of this study was to elucidate the role of the placenta in mediating oxidative stress from mother to offspring. DESIGN: Comparison between normo- and hypercholesterolaemic mothers (n = 36 each) and their children. SETTING: Obstetric wards, hospitals of the University of Naples and Regione Campania. POPULATION: Healthy primiparas delivering by caesarean section. METHODS: Biochemical measurements of oxidative stress and serum leptin in cord plasma and placenta, immunochemistry of placenta microvessels, and vasoreactivity studies were performed. MAIN OUTCOME MEASURES: Oxidative status (i.e. lipid composition and content of oxidised fatty acids, activity of pro- and antioxidant enzymes, immunohistochemical presence of oxidation-specific epitopes) in maternal and cord blood and in placental tissue, as well as vascular reactivity in omental arteries. RESULTS: Hypercholesterolaemia during pregnancy was associated with extensive changes in fatty acid composition of both maternal and cord blood lipids, sufficient to alter vasoreactivity of omental vessels. Results also indicated that the placenta is not only subject to substantial oxidative stress, but that it may further increase fetal oxidative stress through changes of pro- and antioxidant enzyme activities. CONCLUSIONS: The placenta plays an important role in both transmitting and enhancing pathogenic effects of gestational hypercholesterolaemia.

[Immunological abortion: the thyroid factor]. / L'aborto autoimmune: il fattore tiroideo.

Spontaneous miscarriage (SM) is a multifactorial problem involving several couples. Recent studies investigated the correlations between the presence of antithyroid antibodies (ATA) and pregnancy loss, and found that many women with a previous history of recurrent miscarriage, showed high levels of ATA circulating in their blood. Further-more, the thyroid function disorder may also affect the course of pregnancy. Basically, two theories can explain the reasons of the spontaneous termination of pregnancy in presence of ATA: the first theory suggests that the hypofertlity or infertlity of these subjects may be due to a subtle degree of hypothyroidism which is difficult to detect by routine serum hormone determinations; the second theory supports that the presence of thyroid antibodies reveals a more generalized underlying abnormal stimulation of the immune system. Therefore, the thyroid function should be tested before conception and during pregnancy to avoid the pregnancy loss and neuropsychological deficits in infants. Actually, some papers suggest that treatments reserved to women with thyroid antibodies could decrease the miscarriage rate. Unfortunately, there is not agreement about the most effective therapy. We need more large, randomised, placebo controlled, double blind studies.

[Female infertility: the tubal factor]. / Sterilità femminile: il fattore tubarico.

The introduction of medically assisted procreation techniques in the late '70s was an important turning-point for women with tubal infertility and still today the tubal factor, together with the male factor, is the main indication for medically assisted procreation. The outcomes of these in vitro fertilization (IVF) treatments is not significantly different from those of other indications with the exception of women suffering from hydrosalpinx. There is a detrimental effect of hydrosalpinx on the pregnancy rates in women who underwent a IVF cycle and a higher miscarriage rate. It is not yet known which is the mechanism influencing negatively the embryo implantation. However, there is agreement for the surgical removal of the salpinx improving the outcome in assisted reproductive technology (ART), whereas surgical distal tubal repair is the treatment of choice only for patients with preserved tubal mucosa.

FSH: urinary and recombinant.

Infertility affects approximately 10-15% of couples. To enhance the chances of conception, assisted reproductive techniques (ART) have been improved and, in the last 5 years, thousands of babies have been delivered after IVF or ICSI procedures. To obtain an appropriate and controlled ovarian hyperstimulation (COH), clinicians have introduced many drug protocols based on GnRH agonists or antagonists for pituitary down-regulation and FSH or HMG for ovarian stimulation. Currently FSH is the most common drug used for ovulation induction. It is a glycosylated protein with a molecular weight of 28000-30000Da, normally secreted by the anterior pituitary gland. The recombinant preparations of follicle stimulation hormone (r-FSH) are characterized by a higher level of purity, reduced batch to batch variability and no risk of infection. The purpose of this review was to establish the efficacy of different trials based on the use of r-FSH and urinary FSH (u-FSH) comparing published data from randomised studies on the ovulation induction for assisted reproductive techniques. Unfortunately, we did not find any paper with sufficient power to detect a clinically significant difference in pregnancy rates, however, recent meta-analysis among homogeneous trials indicates that the use of r-FSH is to be preferred to u-FSH because more oocytes were collected in cycles with better embryo quality and with more pregnancies. Recombinant FSH has a higher cost per ampoule than urinary FSH, but also a higher effectiveness. A recent cost-effectiveness analysis on follitropin alfa compared to urinary FSH-HP, using a Markov model, confirmed other previous studies that, considering the ongoing pregnancies beyond 12 weeks gestation, r-FSH is the most effective therapy for ovulation induction in ART.

Effects of vaginal progesterone on pain and uterine contractility in patients with threatened abortion before twelve weeks of pregnancy.

Fifty women with previous diagnosis of inadequate luteal phase and threatened abortion underwent a prospective, randomized, double-blind study in one medical center carried out with a parallel trial. The primary objective was to establish the effects of vaginal progesterone (Crinone 8%) in reducing both pain and uterine contractions (UCs). The gel with or without (placebo) vaginal progesterone was administered once a day since the diagnosis of threatened abortion and for 5 days. The efficacy on pain symptom amelioration was evaluated by a 5-score intensity gradation, while the UCs were evaluated by ultrasound. The secondary objective of the study was to evaluate the outcome of the pregnancies. The use of progesterone was effective both on pain relief and on the frequency of the UCs that decreased after 5 days of vaginal progesterone administration (P < 0.005). The evaluation of the ongoing pregnancy and spontaneous abortion in both study groups after 60 days showed that 4 patients of group A and 8 patients of group B miscarried (P < 0.05). In conclusion, patients with threatened abortion benefit from vaginal progesterone by a reduction of UCs and pain. The use of vaginal progesterone improved the outcome of pregnancies complicated by threatened abortion and previous diagnosis of inadequate luteal phase.

Sedation in gynaecologic oncology day surgery.

Gynaecologic oncology day surgery deals primarily with the diagnosis of endometrial, cervical, vulvar and vaginal tumors. Conscious sedation is an important technique used in gynaecologic oncology day surgery. It is often associated with regional anaesthesia. The goals of conscious sedation are to provide effective pain control with complete safety, reduction of the recovery time, of the infection risk and cost. Since the consequences of the immunosuppressive effects of the opiates and the surgical stress could lead to an increased susceptibility to post-operative infections and a possible lack of immunological defence in the cancer patients, we investigated the possibility of eliminating the administration of opiates during minor operations in gynaecologic oncology day surgery. In this study, 400 patients, aged between 35 and 77 years, underwent surgery using sedation at the day hospital annexed to the Gynaecology and Obstetrics Department of the 2nd University of Naples, Italy. The patients were randomized into two equal study groups, according to a computer-generated randomised list. All patients were seen by the consultant anaesthetist three days before surgery. In all cases, during surgery, we monitored the main vital parameters such as ECG, HR and RR, BT, BP and SO. The drugs commonly used were: opioids, hypnotics and benzodiazepines, associated or not, with local anaesthetics. By using these drugs, pain and anxiety were reduced, sedation and amnesia were accomplished. In our experience, conscious sedation seems to be the best choice in gynaecologic oncology day surgery because it presents low toxicity, is short acting and readily reversible, has a low cost and, finally, because it is more comfortable for the patients. Moreover, it is possible to eliminate the administration of opiates during conscious sedation for less invasive surgical procedures.

Human chorionic gonadotropin free beta-subunit in the human seminal plasma: a new marker for spermatogenesis?

UNLABELLED: In the past 20 years, several factors were detected in the human seminal plasma and proposed as markers for spermatogenesis. Human chorionic gonadotropin (hCG) and its beta-subunit were therefore investigated, and their seminal levels were found to be higher than those detected in the serum and to correlate with sperm parameters. OBJECTIVE: We designed a retrospective study to determine the suitability of hCG free beta-subunit concentration in the seminal plasma of fertile and infertile male patients as marker of spermatogenesis. STUDY DESIGN: A total of 79 infertile male patients were divided into four groups by their semen analysis results (group 1 [n=8]: azoospermia; group 2 [n=21]: severe oligozoospermia; group 3 [n=40]: oligoasthenospermia (OAS); group 4 [n=10]: asthenospermia) and 10 healthy volunteers of proven fertility were evaluated. RESULTS: The hCG free beta-subunit levels in the seminal plasma were found to be significantly higher (P<0.0001) in the control group in respect to those assayed in the infertile patients and showed a correlation with sperm count (r=0.5) and total motile sperm density (r=0.5). Twenty-five patients were on treatment with oral Mesterolone (100mg daily) plus Tamoxifen (20mg daily) for 3-6 months. Apart from a significant improvement (P<0.05) in sperm morphology, no significant changes in sperm count and motility were observed after the treatment in all the patients. In the seminal plasma of 10 patients who showed a significant increase in sperm count, hCG free beta-subunit levels were found to be significantly higher compared to those detected in the remaining patients (P<0.01). In all patients, these levels remained unchanged after the treatment. CONCLUSIONS: The evidence regarding the positive correlation between hCG free beta-subunit levels in the seminal plasma and sperm concentration is consistent with the previous results regarding hCG levels. A previous study demonstrated that testosterone levels in seminal plasma correlated with sperm concentrations; from the same evidence regarding hCG we hypothesize that seminal plasma testosterone and hCG levels are correlated. Thus, hCG may play a paracrine role in the intratesticular regulation of testosterone secretion.

[Pregnancy in women with thalassaemia]. / Talassemia e gravidanza.

Women with transfusion dependent thalassaemia suffer from failure of pubertal growth and delayed onset of menarche with amenorrhea, anovulation and infertility. With improved pediatric and hematological care is now possible, for patients with b thalassaemia, to achieve a pregnancy. Pre-pregnancy assessment included checks for hypothyroidism and diabetes, for hepatitis B and C, human immunodeficiency virus, Rubella, cardiac functions, liver functions by estimating aspartate and alanine aminotransferases, gamma-glutamyl transpeptidase, alkaline phospatase, and total plasma proteins. The frequency of blood transfusion needed to be increased in order to maintain the hemoglobin concentration above 10 g/dl. Desferroxamine must be stopped as soon as pregnancy is diagnosed continuing the administration of the folic acid supplements throughout pregnancy. Desferroxamine will be resumed after delivery. The safety of iron chelation with desferroxamine during the periconceptional period and pregnancy has not yet been established. Some animal studies have shown skeletal anomalies; other published studies report seven women with b thalassaemia major who became pregnant while taking desferroxamine: all the women had normal babies. The mode of delivery is usually vaginal, while Cesarean section is performed in those cases with pre-eclampsia, fetal distress, cephalopelvic dysproportion, slow progression of labor, as in women without thalassaemia. In conclusion, with the advent of regular blood transfusion associated with iron chelation therapy, pregnancy in b thalassaemia can be safe for mothers and their babies with appropriate care.

[Correlations between acne and polycystic ovary. A study of 60 cases]. / Acne e ovaio policistico.

BACKGROUND: In this paper we correlated the clinical severity of the polycystic ovary with the clinical manifestation of acne. Acne is an inflammatory process of follicle sebaceous unites. Polycystic ovaries (PCOS) are characterized by several subcapsular cystic follicles with an increased production of androgens. Onset of polycystic ovaries in women with acne, ranges from 45 to 84% of cases. METHODS: Sixty patients, aged from 19 to 28 years, with acne and polycystic ovaries were enrolled. The clinical severity of acne was appraised by millimetric macrophotography. This technique evaluated subsequent phases of acne like comedo, papules or pustules, nodular or scarring lesions. Polycystic ovaries were studied by ultrasound and hormonal assay. Sonora Logic 500 MD, by GE Echography, with 3.5 MHz convex probe, were employed. US examination was performed, in each patient, on day 6th, 10th, 15th and 22th of the menstrual cycle. The following US parameters were considered: dimension of the ovaries, thickness of albuginea and number of cysts. Serum levels of FSH, LH, 17-b-estradiol, progesterone, DHEA-S, testosterone and LH/FSH ratio were determined. Furthermore, the presence of premenstrual syndrome (PMS) was evaluated. RESULTS: We found that in PCOS the increased levels of androgens can cause the onset of acne. Photographic score and ovaries ultrasonography may be useful in the assessment of the clinical association between acne and polycystic ovaries. We observed that the subsequent phases of acne were correlated with the clinical severity of polycystic ovaries and to the presence of Premenstrual Syndrome in 93% of the cases (56 of 60 patients). CONCLUSIONS: US images, compared with visual score, established the clinical association between acne and PCOS in three phases with a progressive severity.

[Influence of ovarian cysts during controlled ovarian hyperstimulation]. / Influenza delle cisti ovariche in corso di iperstimolazione ovarica controllata.

BACKGROUND: The use of gonadotrophin-releasing hormone agonist and purified follicular stimulating hormone increases the incidence of ovarian cysts during in vitro fertilization-embryo transfer cycles. The aetiopathogenesis and the effects of ovarian cysts on controlled ovarian hyperstimulation is unclear; most authors observed no-detrimental influence of ovarian cysts in controlled ovarian hyperstimulation cycles for IVF. The aim of this study was the evaluation of the real incidence and effects of ovarian cysts during in vitro fertilization-embryo transfer cycles. DESIGN: retrospective study. SETTING: reproductive medicine units. PATIENTS: 130 women. INTERVENTIONS: r-FSH--225 U.I. was administered to 130 women i.m. after a long protocol of down regulation with GnRH-a (Triptoreline 3.75). The drug administration was personalized on the basis of: pre-existent patients pathologies; E2 blood levels; ultrasound evaluation of the follicular development. MAIN OUTCOME MEASURES: E2 levels, number and quality of oocytes retrieved, fertilization rate, number of embryos developed, pregnancy rate. RESULTS: At the end of stimulation ovarian cyst was observed in eighteen cases. There was no difference, between the cycles with or without cysts, in the follicular development, number and quality of eggs retrieved, fertilization rate, number and quality of the transferred embryos, pregnancy and miscarriage rate. CONCLUSIONS: The conclusion is drawn that the incidence of ovarian cysts during COH can be reduced observing some simple rules like: to exclude from the stimulation protocols patients with follicles of more than 12 mm diameters as observed at the preliminary ultrasound control; to personalize the therapeutic regime with the pre-existent patients pathologies; ultrasound evaluation of the follicular development; determination of the E2 serum levels, every two days.

[Medical treatment of endometriosis: comparative study of leuprolide acetate and danazol]. / Terapia medica dell'endometriosi: studio comparativo fra il leuprolide acetato ed il danazolo.

Hypo-oestrogenaemia following surgical oophorectomy leads to a regression of the ectopic endometrial tissue. A medical and transitory oophorectomy can be obtained using leuprolide ac., an analogue of LH-RH. In this study we compare the results of therapy with danazol or leuprolide in the treatment of endometriosis. Fifty patients affected by endometriosis were selected laparoscopically and recorded under the classification of American Fertility Society. 20-40 year old women who were admitted had not received any therapy for endometriosis within the previous 12 months. They were allocated at random to open treatment with intramuscular injection of leuprolide (3.75 mg/monthly) or oral danazol. The dose regimen used for this last drug was 600 mg/daily in the majority of patients. 47 women completed a six months therapy, and 42 underwent to a second laparoscopy to evaluate the effects of the treatment. Both danazol and leuprolide were associated to a decreasing of clinical symptoms and to adverse effects that we described in this paper.

[The ovarian hyperstimulation syndrome]. / La sindrome da iperstimolazione ovarica.

The ovarian hyperstimulation syndrome is the most important iatrogenic complication of ovarian stimulation. Every drug used in the treatment of infertility such as FSH, HMG, CC, GnRHa, can lead to the syndrome. The authors of the article report the incidence of OHSS in their patients treated for an assisted conception program; the role of oestrogens, HCG and renin-angiotensin system; the classification, the therapy and how to prevent the syndrome.

[The transplacental therapy of fetal tachyarrhythmias]. / Terapia transplacentare delle tachiaritmie fetali.

The diagnosis of fetal tachyarrhythmia during a routine ultrasound scan imposes a continuous monitoring of fetal heart function through ECG and M-mode echocardiography. Since fetal dysrhythmia can lead to non-immune hydrops or to fetal death a correct diagnosis and an adequate intrauterine therapy are fundamentally important. The authors report the results obtained through a transplacental therapy with digoxin or with digoxin-verapamil in 8 cases with supraventricular tachyarrhythmia. Digoxin was very successful for conversion in sinus rhythm in 4 cases. The combination digoxin-verapamil led to the conversion in only one case resistant to the digoxin therapy.

[Use of triptorelin in the medical treatment of uterine myoma]. / L'impiego della triptorelina nella terapia medica dei miomi uterini.

Following the introduction of GnRH analogue drugs the medical treatment of uterine myomatosis has been proposed. The use of these compounds in fact causes a reduction in mean plasma levels of 17-beta-estradiol which is reflected in the target tissue of myomas. Using this hormone-dependence as a starting Tryptoreline point, the study examined the volumetric changes induced by LH-RH agonist analogue, Tryptoreline, administered to a group of 15 selected patients. The results are encouraging: in all cases there was a marked reduction of tumour volume ranging between 30 and 40%. The therapeutic effect, however, gradually wore off following the suspension of treatment. For this reason, the use of this analogue should be reserved for premenopausal or younger patients for the treatment of menometrorrhagic episodes, or as a preoperative therapy in order to limit blood loss during surgery.

Helicobacter pylori infection in patients with acquired immune deficiency syndrome.

A controlled study was conducted on patients with human immunodeficiency virus (HIV) infection referred for upper endoscopy to evaluate the prevalence of Helicobacter pylori (H. pylori) infection. Four different stains and culture for H. pylori were performed on biopsy specimens from the gastric antrum. Sixteen (40%) of 40 patients with acquired immune deficiency syndrome (AIDS) or AIDS-related complex (ARC) were diagnosed to be infected with H. pylori versus 14 (39%) of 36 age-matched control patients. Eight of 15 AIDS/ARC patients without AIDS-related esophagogastroduodenal findings (53%) were infected with H. pylori versus 8/25 (32%) with endoscopic findings typical of AIDS. No invasion of the lamina propria by H. pylori was noted in any patient. Active chronic gastritis was present in 60% of AIDS/ARC patients and 61% of controls. Fifty-eight and 59%, respectively, of active chronic gastritis cases were infected with H. pylori. All the H. pylori infections, except one, were found in patients with chronic gastritis. In AIDS/ARC patients, H. pylori infection and active chronic gastritis are as common as in other patients referred for upper endoscopy. They may play a pathogenic role, especially when endoscopic AIDS-related findings are lacking. Cell-mediated immune deficiency does not appear to increase the risk of infection with H. pylori.

Fatal toxic epidermal necrolysis during prophylaxis with pyrimethamine and sulfadoxine in a human immunodeficiency virus-infected person.

The combination of pyrimethamine and sulfadoxine (Fansidar) has been reported to cause severe skin reactions including erythema multiforme, Stevens-Johnson syndrome, and toxic epidermal necrolysis. Recently, this drug combination has been used for prophylaxis of Pneumocystis carinii pneumonia in patients with the acquired immunodeficiency syndrome. After two months of weekly prophylaxis with pyrimethamine and sulfadoxine, a 48-year-old homosexual man who was antibody positive for human immunodeficiency virus developed severe widespread erythema, blisters, and loss of skin in sheets, and subsequently died. To our knowledge, this is the first reported case of fatal toxic epidermal necrolysis occurring in a patient with acquired immunodeficiency syndrome-related complex. The lack of absolute safety of prophylaxis with pyrimethamine and sulfadoxine is emphasized in our case, and mandates cautious use and the consideration of less toxic prophylactic measures such as therapy with the recently introduced aerosolized pentamidine.