Molecular analysis of 4q35 rearrangements in fascioscapulohumeral muscular dystrophy (FSHD): application to family studies for a correct genetic advice and a reliable prenatal diagnosis of the disease.

In the majority of facioscapulohumeral muscular dystrophy (FSHD) families (about 95%) the genetic defect has been identified as a deletion of a variable number of KpnI repeats in the 4q35 region, although no specific transcripts from this locus have been isolated so far. Molecular diagnosis is based on the detection by probe p13E-11 of EcoRI small fragments, in the range 10-28 kb, that are resistant to BlnI digestion. In family studies this probe is used with other 4q35 polymorphic markers to assign the haplotype associated with the disease. So far, we performed DNA analysis in 145 FSHD families and identified the 4q35 DNA rearrangement not only in affected individuals, but also in healthy subjects at risk of transmitting the disease, such as non-penetrant gene carriers and somatic mosaics. In addition we applied prenatal tests to 19 fetuses, using DNA extracted from chorionic villi samples (CVS) at 10-11 weeks of gestation. The FSHD status, as determined by the presence of BlnI-resistant small fragments associated with the at risk haplotype, was assessed in nine fetuses; in the remaining 10 cases the disease was excluded. Our results show that molecular analysis of 4q35 rearrangements is a reliable indirect method to perform diagnostic, predictive and prenatal tests in FSHD.

The prognostic significance of acetowhitening of the vulva and HPV-DNA test. A multicentre study.

Acetowhitening of the vulva has been related to a subclinical human papillomavirus (HPV) infection. No consense has been reached about undertaking -or not- any therapy for these acetowhite changes. We have observed from our clinical experience and in a 10 years observational follow-up, that acetowhitening of the vulva regarding high risk (16-18) and low risk (6-11) HPV groups (as assessed by PCR analysis) significantly decreased; and acetowhitening areas negative to polymerase chain reaction (PCR), significantly increased from 53% (202/382) to 85% (276/325) (P<0.001). Our findings suggest that independently from HPV type and in the absence of cofactors, there is a statistically significant spontaneous remission of these areas.

Image cytometry for DNA analysis in endometrial carcinoma correlated with other prognostic parameters.

We evaluated by static cytometry DNA ploidy parameters in 30 stage I-IV endometrial carcinomas and correlated these data with standard clinical-pathological features and disease-free period. We observed a direct correlation between either non-diploid DNA content and deeper myometrial invasion (p<0.02) or D.I.greater than or equal to 1.2 and M2-M3 tumors (p<0.009). The Kaplan Meier survival curves illustrate a more rapid relapse of disease associated with non-diploidy, high 5cExR, high level of proliferation and D.I.greater than or equal to 1.2. while Cox regression model gave relative hazards for disease recurrence of 4, 6, 3.7 and 2.1 for non-diploidy, D.I.greater than or equal to 1.2, high 5cExR and high level of proliferation respectively. This prospective study confirmed the prognostic value of DNA Index, and its usefulness in clinical practice especially in stage I cases, otherwise characterised by favourable standard prognostic factors, is discussed.

Autoimmune thrombocytopenic purpura in pregnancy: maternal risk factors predictive of neonatal thrombocytopenia.

Pregnancy in ATP women is not unusual. The problem of this association concerns the possibility of disease transmission to the fetus due to the crossing of maternal antiplatelet antibodies through the placenta. Maternal risk factors predictive of neonatal thrombocytopenia, can be identified as follows: severe thrombocytopenia, previous splenectomy, high titre of PA-IgG and/or SPB-IgG. In 63 pregnancies in ATP patients, we have evaluated whether the above maternal risk factors, considered in the third trimester, can provide useful criteria for the prediction of neonatal thrombocytopenia. In the third trimester, the distribution of maternal risk factors was as follows: 0 in 7 cases, 1 in 27 cases, 2 in 15 cases, 3 in 12 cases, 4 in 2 cases. From a statistical evaluation, the neonatal platelet values and the maternal risk factors seem inversely correlated (r -0.437; p = 0.0005). In particular, neonatal and maternal platelet count correlated positively (r = 0.249; p = 0.025); moreover, neonatal platelet count correlated negatively with Splenectomy (r = -0.209; p = 0.05), PA-IgG (r = -0.401; p < 0.0005) and SPB-IgG (r = -0.338; p < 0.005). We tried to apply a multiple regression model for all the above parameters which appears statistically significant (p = 0.001); the variability was about 30%. This regression model could be validated if applied to a larger number of cases, and it could represent an alternative to the invasive methods used for the diagnosis of neonatal thrombocytopenia.

Influence of maternal metabolic control and insulin antibodies on neonatal complications and B cell function in infants of diabetic mothers.

Circulating insulin antibodies at birth and the degree of maternal metabolic control were measured in 68 infants of insulin-treated diabetic mothers. Their correlation with neonatal B cell function and with the clinical features of the infants was evaluated in order to better understand their influence on fetal outcome. Maternal metabolic control was assessed on the basis of blood glucose levels, glycosuria and the occurrence of hypoglycemia and/or ketonuria. All infants were clinically evaluated for gestational age, macrosomia, hypoglycemia, hyperbilirubinemia, hypocalcemia, and respiratory distress syndrome. Cord blood plasma glucose, C peptide, and IgG insulin antibodies were also measured. It was shown that poor maternal metabolic control was associated with a higher prevalence of fetal morbidity as well as with signs of B cell hyperfunction. Also the presence of circulating insulin antibodies correlated well with higher C peptide levels and with several neonatal complications. B cell hyperfunction, indicated by high C peptide levels in the infants of diabetic mothers, may possibly play a causal role in the pathogenesis of fetal morbidity. In conclusion, a good fetal outcome in insulin-treated diabetic pregnancies was associated with and may have depended upon: (1) good maternal metabolic control, and (2) absence or low levels of circulating insulin antibodies.

DNA ploidy and HPV subtypes in cervical smears of HIV-sero-positive and negative patients.

Epidemiological studies have demonstrated that cervical HPV infection and precancerous lesions of the cervix are more common in HIV-seropositive patients. However little is known about the natural history of these lesions in this population. In the present study cervical smears from 36 patients, 18 HIV-seropositive women and 18 matched controls were evaluated with the aim of quantifying morphological alterations and to evaluate DNA ploidy and HPV subtypes. Cervical lesions in HIV-seropositive patients were diploid in 50% of the cases compared to 25% in controls. The only HPVs identified by ISH were types 16/18 and no significant differences were observed in the control population. In contrast, cytological evidence of HPV infection and dysplastic changes was greatly increased in smears from HIV patients compared to HIV-seronegative women. Less than 5% of the cells showed HPV associated changes in controls while 10% to 30% of the cells were affected in HIV-patients. We suggest that the Papanicolaou test should be effective for detecting cervical disease and for a close follow-up of this population. Moreover, while additional studies with larger population groups and different population bases are needed, these findings are suggestive of the possible use of morphological criteria for the identification of HIV-seropositive subjects.

Use of low-frequency electrical impedance measurements to determine phospholipid content in amniotic fluid.

In this report we propose a new method for an in vitro test of the foetal lung maturity based on the measurement of the electrical conductivity of the overall amniotic fluid obtained from transabdominal amniocentesis, since this quantity can be linked to a first approximation in a very simple way to the phospholipid content. We have carried out measurements of 85 different samples of amniotic fluid as a function of gestation weeks and we have observed a pronounced change of the electrical conductivity that reflects the increase in the phospholipid concentration occurring at the end of normal pregnancies. The method could be further developed to obtain similar information on in vivo experiments by means of bioelectric impedance tomography, taking advantage of the frequency dependence of the tissue electrical impedance.

Autoimmune thrombocytopenic purpura: maternal and fetal disease.

We have followed up 63 pregnancies in women with autoimmune thrombocytopenic purpura (ATP). Of these, 15 were previously splenectomized. The characteristics of the sample can be summed up as follows: average age 27 years (17-41); platelets at the beginning of pregnancy, mean 129.5 x 10(9)/l (range 16-488); platelets at delivery, mean 133 x 10(9)/l (range 8-477); PA-IgG at delivery, mean 320 ng IgG/10(7) platelets (range 10-1000); SPB-IgG at delivery, mean 262 ng IgG/10(7) platelets (range 10-1000). There were 30 spontaneous deliveries and 33 cesarean sections. Forty-two newborns had a platelet count within the normal range while nine had a platelet count less than or equal to 150 x 10(9)/l, while six had less than or equal to 100 x 10(9)/l and a further six less than or equal to 50 x 10(9)/l. The aim of this study is the evaluation of maternal risk and of possible feto-neonatal thrombocytopenia at birth. In this regard, the following parameters were considered: previous maternal splenectomy; the platelet count at the beginning of pregnancy; the platelet count and the titres of PA-IgG and SPB-IgG at delivery. Preliminary statistical evaluation of these parameters enabled us to identify a risk score. From this it was possible to obtain an optimum management of the final stage of pregnancy regarding the therapeutic approach and the timing of delivery.

Activated T cells in normal pregnant women and neonates.

The phenotyping of T-cell subsets and T cells at different stages of activation was performed using a panel of monoclonal antibodies in samples from normal pregnant women at different stages of gestation and in the cord blood of neonates. The data obtained from pregnant women showed a slight decrease in the total number of T cells at the beginning of pregnancy, whereas there was a clear increase in 4F2-positive lymphocytes after a few months of gestation. No significant increase in Class II-positive lymphocytes was observed in normal pregnant women in comparison with adult healthy women. The data from neonates revealed a clear decrease of OKT3- and OKT4-positive cells and an increase of 4F2-positive cells in comparison with control subjects. These data indicate that alerted, but not fully activated, lymphocytes are present in the circulation of both the mother, after the first months of pregnancy, and the neonate. This finding reinforces the concept that during pregnancy there is an activation of certain immune components rather than a general depression of the immune system.

Management of primary megaureter in infancy.

The natural history of primary megaureter was evaluated in 26 infants (20 boys, 6 girls) with 38 nonrefluxing primary megaureters. Prenatal ultrasound diagnosis had been made in 21 infants. In all patients, primary megaureter was associated with pelvicaliceal dilatation. Two patients with urinary tract infection and two with agenesis of the contralateral kidney underwent surgical treatment. Asymptomatic infants with good differential renal function on DTPA scanning were managed conservatively regardless of their postfurosemide drainage curve. One of these infants later developed a slight deterioration in function and was submitted to surgery. One patient was lost to follow-up. Twenty patients, after a period ranging from 4 months to 8 years, showed partial or complete regression of ureteral dilatation with stable good renal function. In conclusion, many asymptomatic primary megaureters in infancy improve spontaneously and do not require surgical treatment.

Management of human papilloma virus vulvo-perineal infection with systemic beta-interferon and thymostimulin in HIV-positive patients.

OBJECTIVES: To evaluate the effect of a combined therapy based on beta-interferon and thymostimulin in HIV-seropositive women with florid vulvoperineal condylomatosis. METHODS: Nineteen HIV-seropositive women affected by HPV florid vulvo-perineal infection were studied. Their ages ranged from 19 to 32 years (mean = 26); all were smokers; 11 (57.89%) were intravenous drug users with a mean period of drug addiction of 5 years. No patient was on AZT therapy. All patients underwent a combined medical therapy based on beta-interferon administration with doses of 3,000,000 IU i.m. daily for 7 days and subsequently on alternate days for 2 weeks associated with the administration of 70 mg i.m. of thymostimulin per day on alternate days for 30 days. RESULTS: Seven (36.84%) patients showed complete recovery at the end of follow-up; 5 (26.31%) patients showed partial recovery and 7 (36.84%) did not respond to therapy. CONCLUSION: A pro-host immunotherapy seems to be indicated in patients affected by immunodeficiency syndrome.

HPV and intraepithelial neoplasia recurrent lesions of the lower genital tract: assessment of the immune system.

OBJECTIVE: To evaluate the immune state in patients with genital relapse HPV and intraepithelial lesions of the lower genital tract. METHOD: Forty-three patients were selected. Twenty-one were affected by recurrent HPV infection either alone or combined with intraepithelial neoplasia treated by laser surgery, and 22 had been previously-treated and clinically cured without recurrence during a follow-up from 18 to 24 months. The diagnostic protocol included colposcopy with eso- and endocervical cytology histologically confirmed by directed biopsy. Afterwards patients underwent a systemic immunogenic evaluation. RESULTS: NK cell reduction was strictly related to HPV infection associated with intraepithelial lesions; B-lymphocyte reduction was percentually greater in patients affected by HPV alone; activation of R-IL2 increased in a percentage overlapping in the two groups indicating patient reaction to the virus. CONCLUSION: Our study supports the theory that immune response directed against viral antigens is one of the most important effectors in the control of HPV infections and that HPV is the cause of a systemic rather than local lesion.

Abdominal cystic lymphangioma in a woman at 14 weeks' gestation: case report.

Lymphangiomas are uncommon benign tumors that most commonly present early in life, and are even more exceptional in adults. We present a rare case of a 14-week pregnant woman who had a large septated cystic lesion adherent to the ileum. Despite analysis by ultrasound, the correct diagnosis was established only via laparotomy and she had surgical resection of an abdominal lymphangioma.

Multiple primary neoplasms. Considerations on 42 cases.

During the years 1977-1993, 2,108 cases of gynecologic primary neoplasms were observed in the Department of Gynecology and Obstetrics of La Sapienza University in Rome. Forty-two cases were multiple primary cancers. Of these, 27 had multiple primary neoplasms involving only gynecologic sites, 15 had a neoplasm involving also another site; 3 patients had triple primary neoplasms. The most frequent neoplasm associations observed in our case series were: breast-ovary; endometrium-ovary; breast-cervix; endometrium-bowel (sigma-colon).

Cervical intraepithelial neoplasia and bacterial vaginosis: correlation or risk factor?

Bacterial Vaginosis (B.V.) is a syndrome defined microbiologically where lactobacilli-dominated flora is exchanged with an abundant complex flora dominated by strict and facultative anaerobic bacteria, constituted by gardenerella, micrococci, streptococci and staphylocci. It has been suggested that B.V. could be important in the development of cervical intracpithelial neoplasias, because the abnormal micro flora can produce careinogenic nitrosamines. The aim of this investigation was to evaluate the correlation between CIN and B.V. in 1,008 patients (median age: 28 years, range 17-60) who were divided into two groups: the first group consisted of 504 patients affected by CIN of different degrees, the second of 504 patients without CIN. All patients were submitted to colposcopy, pap-tests, cytology of the cervical canal and microcolpohysteroscopy in the case of lesions invading the cervical canal. The diagnosis of B.V. is based on four criteria: presence of clue cells, pH 4.5, positive amine test and increased vaginal discharge. X2 Pearson analysis was applied for statistical evaluation of the data. Among the 504 patients in the first group (women affected by CIN of different degrees), 180/504 (36%) women presented B.V. Among the second group (504 women without CIN), 248/504 (49%) women presented B.V. The results of our study demonstrate that there is no significant correlation between CIN and B.V. In the first group of patients affected by CIN and B.V., B.V. was present in 36% of the cases, while in the second group of patients not affected by CIN. B.V. was present in 49% of the cases (P < 0.00005).

Correlation between cervical intraepithelial neoplasia and human papillomavirus male infections: a longitudinal study.

BACKGROUND: Genital HPV infection is one of the most common sexually-transmitted diseases. The aim of the study was to evaluate the correlation between HPV-associated lesions in male partners of women affected by CIN. METHODS: 210 male partners of women affected by CIN were examined in a long-term follow-up (from 5 to 13 years). The diagnosis in females was performed by cytology, colposcopy and histology. Male partners were submitted to clinical examination, peniscopy and biopsy. RESULTS: 111/210 (53%) females had CIN I, 53/210 (25%) and 46/210 (22%) had CIN III. Subclinical lesions were associated with 18%, 28% and 24% of male partners of women with CIN I, CIN II and CIN III, respectively. Clinical lesions were observed in 7% and 10% of sexual partners of women with CIN I and CIN II, respectively. Mixed lesions affected only 3% of sexual partners of women with CIN I. CONCLUSIONS: Our data show that the transmissibility of HPV infection to the male partners of women affected by CIN was easier when there was a lower grade of CIN. In fact, male partners of women with CIN III had a lower percentage (26%) of clinical or subclinical HPV skin lesions.

Diagnostic methods of early detection of endometrial hyperplasia and cancer.

Endometrial carcinoma is the most frequent genital neoplasia in women. Our own experience is based on a group of 3310 patients who were studied between 1985 and April 1994. In 2148 cases, equivalent to 56.4%, hysteroscopy was performed because of the presence of abnormal uterine bleeding (AUB). We found 672 endometrial hyperplasia (601 low risk-71 high risk) and 93 endometrial carcinoma. According to our experience we can assert that: hysteroscopy allows the measurement of the extension of the intracavitary neoplastic pathology and the definition of its topographic map; it is the only method that allows a target biopsy; it facilitates the evaluation of the extension of the neoplasia in the cervical canal, with the help of a deep biopsy into the cervical tissue (as proposed by La Sala) increasing its diagnostic accuracy.

[Prenatal diagnosis of osteogenesis imperfecta. Report of a case]. / Diagnosi prenatale di osteogenesi imperfetta. Descrizione di un caso.

Osteogenesis imperfecta is a heterogeneous group of connective tissue disorders; it is characterized by blue scleras, bone fragility and deformity secondary to recurring fractures. A case of osteogenesis imperfecta personally observed at the 21st week of gestation is presented.

[Diabetes and pregnancy: physio-pathogenetic and epidemiologic aspects]. / Diabete e gravidanza: aspetti fisio-patogenetici ed epidemiologici.

Diabetes mellitus is one of the most common maternal illnesses resulting in congenital malformations. All complications of pregnancy, either with diabetes pregestational or gestational, are directly or indirectly related to the degree of metabolic control. If it is not treated properly, diabetes in pregnancy causes major problems for both mother and fetus. The only way to reduce complications to the minimum and locate them near to those of the normal population, is to achieve a good metabolic control. Multi-disciplinary approach in which obstetricians, physicians, paediatricians are involved, combined with intensive monitoring and therapy throughout pregnancy, could achieve successful results in women with complicated diabetes. This objective is subordinate to early diagnosis for gestational diabetes and planning of pregnancy for diabetic women.

Acute fatty liver of pregnancy. Description of a case of survival.

The Authors describe a case of acute fatty metamorphosis of the liver in pregnancy in a patient who survived and delivered a living female child. Clinical and istopathologic features, possible etiologic factors and possible treatment for the mother and fetus are discussed. The Authors, finally, think that acute fatty liver of pregnancy is more frequent than expected and that it should be considered in all cases of jaundice in pregnancy.