Soluble HLA-G molecules in follicular fluid: a tool for oocyte selection in IVF?

Currently, different approaches are used to select oocytes for in vitro fertilization (IVF) procedures, but they do not assure a significant association with the pregnancy outcome. Since several studies have proposed the expression of HLA-G antigens in early embryos to be a possible marker of elevated implantation rate, we have investigated the presence of soluble HLA-G molecules in 50 follicular fluids (FFs). The results have shown soluble HLA-G antigens (sHLA-G) in 19/50 (38%) FFs. Furthermore, we have related the presence of sHLA-G molecules in FFs to detection of the soluble antigens in culture supernatants of the corresponding fertilized oocyte, evidencing a significant relationship (p=1.3 x 10(-6); Fisher exact p-test). Specific ELISA and Western blot approaches identified both HLA-G5 and soluble HLA-G1 molecules in FFs while immunocytochemical analysis indicated polymorphonuclear-like and granulosa cells as responsible for production of sHLA-G1 and HLA-G5 molecules. In contrast, only sHLA-G1 antigens were detected in culture supernatants of fertilized oocytes. Overall, these results suggest a role for sHLA-G molecules in the ovulatory process and propose the FFs analysis for sHLA-G molecule presence as a useful tool for oocyte selection in IVF.

Evaluation of clinical efficacy of three different insemination techniques in couple infertility. A randomized study.

AIM: The aim of the study was to compare the clinical results and efficiency of three insemination technique: intraperitoneal insemination (IPI), fallopian sperm perfusion (FSP) and intrauterine insemination (IUI). METHODS: The experimental design was a prospective, randomized trial. A total of 101 homologous insemination cycles were performed in 71 consecutive couples with unexplained or male subfertility. Couples were randomized to receive IPI or FSP or IUI by predefined tables of randomization and each couple was submitted to the same insemination technique. The primary outcome of the study was the achievement of clinical pregnancy. RESULTS: The results of the study underlined firstly that basal couple composition was not statistically different between the three groups. Moreover, no significant difference in clinical pregnancy rate was observed, despite a clearly positive trend for FSP, especially for unexplained infertility. CONCLUSIONS: Our results showed that the three techniques of insemination IUI, FSP and IPI have similar efficacy on the achievement of clinical pregnancy in couples affected by longstanding infertility.

[Pattern of normal fetal heart rate in the third trimester of pregnancy]. / Il profilo della frequenza cardiaca fetale media nel corso del terzo trimestre di gravidanza.

BACKGROUND: It has been shown by previous studies that the mean fetal heart rate (FHR) decreases from the 11th to the 30th week, afterwards the values remain more stable. In the present study we evaluated the FHR pattern throughout the last stages of pregnancy and particularly we estimated the interindividual variability to achieve the maturation of the parasympathetic nervous system in the control of the FHR rythm. METHODS: Serial FHR determinations were made from the 28th week until term in 126 physiological pregnancies, submitted to umbilical Doppler velocimetry. The mean FHR was measured on the length of 6 cardiac beats. The FHR patterns of 48 healthy pregnancies, were also analyzed longitudinally. RESULTS: The transversal data, related to the 126 physiological pregnancies, show a further decrease of the FHR during the last stages of pregnancy: from the 28th to 40th week it drops from the average value of 141 bpm to 133 bpm (r = -0.1469). The longitudinal data show that this decrease occurs in 27% of patients between the 30th and the 33rd week and in 73% of patients between the 34th and the 40th week. CONCLUSIONS: The conclusion is drawn that the physiological decrease of the mean FHR value occurs for each fetus at different gestational ages due to a different balance between the sympathetic and parasympathetic tone; moreover, we find that the interindividual variability in achieving the full maturation of the parasympathetic nervous system is quite wide.

[Effectiveness and tolerability of oral liquid ferrous gluconate in iron-deficiency anemia in pregnancy and in the immediate post-partum period: comparison with other liquid or solid formulations containing bivalent or trivalent iron]. / Efficacia e tollerabilità del gluconato ferroso orale liquido nell'anemia da carenza da ferro in gravidanza e nell'immediato post-partum: confronto con altre formulazioni liquide o solide contenenti ferro bivalente o trivalente.

UNLABELLED: From the early months of pregnancy and even more so later, women suffer a deficiency of iron along with a decline in their red blood cell count. It is also now clear that women who take iron supplements during pregnancy do not suffer the same post-natal reduction in hemoglobin and ferritin as those who don't make it. A study was therefore conducted on 40 women aged 20-35, with iron-deficiency anaemia during or immediately after pregnancy all of whom presented Hb < 10 gr/dl, Ht < 33% and serum iron < 60 micrograms/dl. All women with pregnancy-related pathological conditions, pre-existing on concomitant disease (Type I diabetes, heart diseases etc.) were excluded from the study. The women whose blood chemical parameters were largely homogeneous at the start of the study were divided into four treatment groups of 10 patients each and were treated as follows: Group A with oral liquid ferrous gluconate (75 mg per diem in 2 vials a day); Group B with solid ferrous gluconate (80 mg per diem in a single effervescent tablet); Group C with solid ferrous sulphate (105 mg per diem in a single tablet); and Group D with ferric protein succinylate (80 mg per diem in 2 vials a day). All were given iron treatment for 30 days. Treatment efficacy was analysed by comparing basal and final parameters using the T-test for paired dependent samples. The tolerance of the 4 treatment protocols was assessed by the analysis of any side effects such as nausea, vomiting, epigastric pain, diarrhoea, constipation or other disorders reported by patients during treatment. RESULTS: Analysis of the therapeutic efficacy parameters (red blood cells, hemoglobin, hematocrit and serum iron) showed significant improvements but no statistically significant differences between the groups. However, the Group A patients treated with oral doses of liquid ferrous gluconate received a significantly lower cumulative dose of iron elements than the other groups: in detail 150 mg (p < 0.05) less than Groups B and D; 900 mg (< 0.001) less than Group C. By the end of treatment the Group A patients revealed significant increases versus basal values in red blood cells (p < 0.001) 1,051,000 per mm3 or 33%, in Hb (p < 0.001) 2.83 gr/dl or 32%, in Ht (p < 0.001) 8.32% or 32%, in serum iron (p < 0.05) 19.5 micrograms/dl or 61%. The same group also showed an increase in Ferritin amounting to 7.8 micrograms/dl or 24% of the basal value. As to safety, only Group A patients reported no side effects and produced no drop-outs. Gastrointestinal and other aspecific side effects caused 1 drop-out each in Groups B and C and 2 drop-outs in Group D. CONCLUSION: Numerous preparations containing bivalent or trivalent iron are available for the treatment of iron-deficiency anaemia during or immediately after pregnancy. It has been shown that preparations containing ferrous salts (+2) are more easily absorbed than those containing ferric salts (+3) since the former can be immediately absorbed by the duodenal mucosa. The study reported here reveals that oral ferrous gluconate in liquid form is more effective and above all better tolerated than other solid or liquid formulations containing elementary iron.

[Maternal-fetal transmission of the acquired immunodeficiency virus. Authors' experience]. / Trasmissione materno-fetale del virus da immunodeficienza acquisita. Nostra esperienza.

UNLABELLED: The determination of human immunodeficiency virus (HIV) status of the newborn remains a major diagnostic problem as a routine test, which detects antibodies to HIV, is of limited value in evaluating newborns. However, the risk of infection for a baby whose mother is an HIV carrier is not yet clear. OBJECTIVE: A prospective study to evaluate maternal transmission of HIV in our population. In order to provide a better estimate of the effect of some of the risk factors associated with HIV transmission we present a stratification of the transmission rate by these factors. STUDY DESIGN: Between January 1990 and December 1994 were examined a total of 10.949 pregnancies. To evaluate maternal HIV transmission to the fetus we considered a positive routine diagnostic test, which detects antibodies to HIV, of definite significance only at the age of 18 months or over. RESULTS: Our prospective study shows an overall transmission rate in our population of 31.25%. Maternal disease stage, as reflected by CD4+ cell count, prematurity, mode of delivery (episiotomy) was correlated with HIV transmission. CONCLUSIONS: Although some risk factors have been recognized, our understanding of the various modes of transmission is still limited. We conclude from our experience and previous studies that the risk of maternal to newborn transmission of HIV must be determined according to the specific characteristics of each parturient population.

Fractal analysis: a new method for evaluating fetal heart rate variability.

In this report we describe how the technique of "deterministic chaos" can be successfully applied to the study of fetal cardiotocography (CTG). The CTG tracings were analysed, after identification as "normal" or "pathological" by experts and by a computerised system, to evaluate the correlation dimension (D2), and the graphic analysis of the attractors which visualize the phase-space of the system that generated the signal (of the state of the underlying hypothetical system). Thanks to this analysis of the traces, it is possible to detect the existence of chaos in the system that generates a certain signal in this case the fetal heart rate. Normal CTG traces at term (37-41 wks) show a fractal dimension (D2) index ranging from 4.22 to 5.22; highly pathological CTG tracings show a D2 ranging from 2.0 to 3.09. Moreover, we introduced recurrence plots in the study of CTG, as tool for non stationary signals, working in real time. To our knowledge this is one of the first attempts to apply fractal analysis and recurrence plots in the evaluation of fetal heart rate in pregnancy, using very small data sets.

CA125 in culture medium of preimplantation embryo.

The first stage of the implantation is the adhesion of the embryonic pole of the blastocyst to the decidua. Such a phenomenon has been demonstrated to be dependent on the presence of glycoproteic compounds, produced partly by the decidua and partly by the embryo. CA125 is an antigenic determinant associated to a glycoprotein expressed by various embryonic tissues. The objective of our research has been to measure the production of CA125 by the embryo in the initial phase of its development. Patients were recruited from our in vitro fertilization program. The culture medium used for the oocytes and for the embryos was collected and CA125 levels were measured. The results indicate that there is not a statistically significative difference between the values of CA125 measured in the mediums where a pronucleus or an embryo was present and the negative controls. From our data, therefore, it can be concluded that CA125 expression begins later in the human embryonic development than 8-cells-stage embryo.

Chorion villosum does not express progesterone and estrogen receptors during the first trimester of pregnancy.

First-trimester chorion villosum plays the key role in the development of human placenta and secretes a large number of hormones and hormone-related substances. To test whether progesterone and estrogen could have regulatory effects on first-trimester chorion villosum functions, we investigated the presence of progesterone receptor (PR) and estrogen receptor (ER) in 47 chorion villosum samples from patients who underwent abortion during the first trimester of pregnancy. The study was carried out using immunohistochemical methods. No ER and PR positivity was evidenced in the 47 chorion villosum samples examined. The possibility that progesterone and estrogen play a role in the regulation of chorion villosum functions should therefore be excluded.

Maternal serum CA 125 levels in first trimester abortion.

OBJECTIVE: To assess the source of maternal serum CA 125 during the first trimester of pregnancy. STUDY DESIGN: CA 125 was measured in stored samples from nonviable pregnancies of 8-13 weeks gestation. The study group comprised 19 women with vaginal bleeding and 13 non-bleeders. Only patients in whom chromosome analysis of the products of conception demonstrated a normal caryotype were included. CA 125 levels were expressed in multiples of the median (MoM) for normal pregnancies of the same gestational age. RESULTS: Median MoM values of CA 125 were significantly higher in women with vaginal bleeding (1.81 MoM) as compared both to non-bleeders (0.82 MoM; p < 0.01-Mann-Whitney U-test) and to the normal pregnancies (1.01 MoM; p < 0.05). No significant difference was found between non-bleeding women and controls. CONCLUSIONS: The present study indicates that in non-viable pregnancies with euploid fetuses an increase in maternal serum CA 125 levels was found only in presence of decidual disruption associated to vaginal bleeding. These findings are compatible with a prevalent decidual source of this antigen.

Amniotic fluid levels of CA 19.9 and CA 15.3 in normal and Down's syndrome pregnancies.

CA 125, CA 19.9 and CA 15.3 are antigenic determinants on glycoproteins commonly used as serum markers in gynecologic oncology. CA 125 was also investigated in the amniotic compartment of Down's syndrome pregnancies. The objective of this study was to evaluate the concentration of CA 19.9 and CA 15.3 in the amniotic fluid of normal and Down pregnancies. The concentration of these antigens was measured in stored amniotic fluid samples from 15-19 weeks pregnancies with and without Down's syndrome fetuses and expressed in multiples of the median for normal pregnancies of the same gestational age. The study group included 20 samples selected from single Down's syndrome pregnancies. Each of these samples were matched with three control samples from unaffected pregnancies, matched for maternal age, gestational age and duration of storage of the sample. The median MoM values of CA 19.9 and CA 15.3 in Down's syndrome pregnancies were respectively 1.11 MoM and 1.16 MoM, not significantly different from those of unaffected pregnancies (CA 19.9: 1.02 MoM; CA 15.3: 0.99 MoM). In the present study we have not found significantly different levels of CA 19.9 and CA 15.3 in the amniotic fluid of fetuses affected by Down's syndrome and in the controls.

Decidual progesterone and estrogen receptors in the first trimester of pregnancy.

Receptor content of human decidua in early pregnancy (weeks 6-12) was investigated. Fifty-three tissue samples were obtained from voluntary patients undergoing abortion and whose gestational age range from 6 to 12 weeks. Blood samples were drawn at the time of operation in order to measure circulating estradiol (E) and progesterone (P) concentrations. Tissue samples underwent first histological confirmation and then were analyzed for receptor content by immunohistochemistry (IH) and by the conventional ligand binding technique (LBA). Estrogen receptors (ER) appeared to be always undetectable by IH (53 samples). LBA measured a significant amount of ER (> 10 fmol/mg) in two samples, borderline values (3-10 fmol/mg) in 6 and no binding in the other three. No relation was apparent between PR levels and either gestational age or blood P concentration. ER were possibly downregulated by the high E levels, and their synthesis inhibited by the high P levels.

Abnormal amniotic fluid levels of CA 125 in second-trimester Down syndrome pregnancies.

The aim of this study was to evaluate the concentration of CA 125 in second trimester amniotic fluid from Down syndrome pregnancies. CA 125 was measured in stored amniotic fluid samples from pregnancies of 14-19 weeks' gestation with and without Down syndrome fetuses. CA 125 levels were expressed in multiples of the median (MOM) for normal pregnancies of the same gestational age. Twenty-one pregnancies with Down syndrome fetuses and 63 unaffected controls matched for maternal age, gestational age, and duration of storage were studied. The median MOM values of the affected pregnancies were significantly higher than those of the controls (1.41 MOM versus 0.99 MOM). These findings show that there is an increased concentration of CA 125 in second-trimester amniotic fluid from Down syndrome pregnancies.

[Cardiotocographic monitoring of fetal health. Comparative evaluation of traditional and computerized methods]. / Monitoraggio cardiotocografico del benessere fetale. Valutazione comparativa fra la metodica tradizionale e quella computerizzata.

One hundred physiological pregnant women and 76 pathological pregnant women suffering from gestational diabetes and pregnancy-induced hypertension underwent a cardiotocographic examination during the course of routine diagnostic tests. The interpretation of cardiotocographic printouts was carried out using traditional as well as computerised methods. The outcome of these tests was then related to neonatal outcome and other parameters which contributed to defining the prognosis of pregnancy. Computerised analysis was found to provide a more reliable diagnosis in comparison to traditional methods in identifying those pregnancies with a pathological neonatal outcome. In particular, in the group of physiological pregnancies, computerised interpretation proved more reliable in 87.5% of cases in which neonatal outcome was pathological; on the contrary, traditional interpretations only revealed 37.5% of the same cases. In pathological pregnancies, automatic interpretation was also found to be more reliable in predicting the non-pathological outcomes, whereas traditional methods provided a high incidence of uncertain answers.

[Hematocrit and hemoglobin, parameters of hematic viscosity, in pregnancy-induced hypertension]. / L'ematocrito e l'emoglobina, parametri di viscosità ematica, in corso di ipertensione gravidanza-indotta.

During pregnancy because of physiologic hemodiluition and changes in various plasma protein levels, plasma viscosity is decreased compared to the non pregnant condition. Specifically the whole blood viscosity profile throughout pregnancy follows that of the hematocrit. However some pathological condition like pregnancy induced hypertension and intrauterine growth retardation are characterized by an increase of plasma viscosity. In order to evaluate the effect of plasma viscosity on placental perfusion, in 41 patients affected by pregnancy induced hypertension and with no iron deficiency we compared maternal hemoglobin and hematocrit to the birth weight. High maternal hemoglobin and hematocrit levels were associated to an increased frequency of low weight for date newborns (less than or equal to 10th centile), although the relationship with the hemoglobin levels is stronger (p less than or equal to 0.02) than the one with the hematocrit (p less than or equal to 0.05). In contrast, high weight for date newborns (greater than or equal to 90th centile) were not related to maternal hemoglobin and hematocrit parameters. We found that maternal hemoglobin and hematocrit, indicators of plasma viscosity, are useful in predicting low birth weight, but not high birth weight. We speculate that hypoxia due to the modification of microcirculation is a very important factor in determining the low birth weight; in contrast the oxygen is not the only factor involved in determining the high birth weight.

Clinico-pathological profile of endometrial carcinoma in young women (under 40 years of age).

Twenty primary endometrial carcinomas in 40 year-old or younger women were collected from a series of 845 carcinomas and newly re-evaluated. The typical histological profile was well-differentiated adenocarcinoma (30%) or adenoacanthoma (35%); however, a case of adenosquamous carcinoma and an exceptional malignant mixed mesodermal tumor were also observed. No tendency for myometrial infiltration (M0) was displayed by 11 (55%) of the cases, while 6 were M1 and 3 were M2 infiltrating neoplasias. Adenomatous hyperplasia was associated with cancer in 53% of the 15 assessable cases. Histopathological diagnostic problems were elicited in 5 cases only. Typical Stein-Leventhal syndrome occurred in 7 patients (35%), while polycystic ovaries were observed in 15 cases (75%). Usually, the patients were obese (40%) and nulligravida (65%). All but one case were stage I disease. Prognosis seems to be excellent following surgery.

Monoclonal antibodies in serological, immunoscintigraphic and immunocytochemical diagnosis of epithelial ovarian cancer.

In present work the Authors examine the utility of CA 125 assay in patients affected by epithelial ovarian cancer. It is stressed that marker negativity does not necessarily imply the absence of disease. On the other hand the Authors underly the almost total absence of false positives, the only problem being with cases of benign gynaecological pathologies. The CA 125 assay appears to provide informations in the evaluation of response to therapy. Finally they note two more recent applications of monoclonal antibodies: first, marking these substances with radioisotopes; second, immunohistochemistry that shows the presence and location of the antigen at the histological level. (By editorial staff).

Levo-norgestrel-nova-T and precancerous lesions of the endometrium.

The authors report the preliminary data on the topical treatment of endometrial hyperplasia by an intrauterine device releasing Levo-norgestrel (LNG-Nova-T), in a series of 31 patients. The patients examined so far by endometrial biopsies performed at predetermined intervals showed a complete histological regression of the hyperplasia, regardless of its pattern.

Dysgerminoma of the ovary: a report on 29 patients.

Twenty nine patients with pure dysgerminoma of the ovary were referred to the Institute of Radiology of the University of Florence between 1960 and 1983. Twenty six patients received radiation therapy after surgery; three cases, classified as Stage IA, were not submitted to postoperative radiation therapy. Five patients relapsed 2 to 16 months after initial surgery, and three of them died because of their tumours. Sites of relapse were found to be outside the treated volume in four out of these five patients. The overall survival rate for this series is 90%. The survival rate seems to depend not only on the stage of the disease, but also on initial accurate staging and appropriate therapy. Criteria for selecting patients from whom post-operative radiotherapy may be withheld are discussed.

Perioperative chemotherapy in the treatment of carcinoma of the uterine cervix.

Two hundred forty-eight women from 1969-71 who underwent surgery for carcinoma of the cervix and were treated with triaziquone (0.2 mg i.v. daily) from day 2-5 after the operation, are compared to a group of 301 conventionally treated patients examined over the 3 following years. Survival rates after 5 years did not differ significantly between the 2 groups of patients, considered either globally or divided according to their clinical or surgical-pathological stage. The incidence of complications in the "treated" patients was not significantly higher, particularly considering local and general post-operative recovery. A prospective randomized study using drugs with a high cytotoxic activity in patients with tumors limited to the cervix and unfavorable prognostic factors, could in the future help evaluate the possibility of using surgical adjuvant chemotherapy in carcinoma of the cervix.