A phase-III prevention trial of low-dose tamoxifen in postmenopausal hormone replacement therapy users: the HOT study.

BACKGROUND: Postmenopausal hormone replacement therapy (HRT) relieves menopausal symptoms and may decrease mortality in recently postmenopausal women, but increases breast cancer risk. Low-dose tamoxifen has shown retained activity in phase-II studies. METHODS: We conducted a phase-III trial in 1884 recently postmenopausal women on HRT who were randomly assigned to either tamoxifen, 5 mg/day, or placebo for 5 years. The primary end point was breast cancer incidence. RESULTS: After 6.2 ± 1.9 years mean follow-up, there were 24 breast cancers on placebo and 19 on tamoxifen (risk ratio, RR, 0.80; 95% CI 0.44-1.46). Tamoxifen showed favorable trends in luminal-A tumors (RR, 0.32; 95% CI 0.12-0.86), in HRT users <5 years (RR, 0.35; 95% CI 0.15-0.82) and in women completing at least 12 months of treatment (RR, 0.49; 95% CI 0.23-1.02). Serious adverse events did not differ between placebo and tamoxifen, including, respectively, coronary heart syndrome (6 versus 4), cerebrovascular events (2 versus 5), VTE (2 versus 5) and uterine cancers (3 versus 1). Vasomotor symptoms were 50% more frequent on tamoxifen. CONCLUSIONS: The addition of low-dose tamoxifen to HRT did not significantly reduce breast cancer risk and increased climacteric symptoms in recently postmenopausal women. However, we noted beneficial trends in some subgroups which may deserve a larger study.

B lymphocyte subsets and their functional activity in the early months of life.

In the present study we evaluated B-cell subsets and their functional development in 74 newborns from birth to 6 months of life. Moreover, we evaluated natural antibody production in vitro. The results documented a predominance of naive B-lymphocytes at all time-points evaluated, decreasing from birth to 6 months (p=0.009). The percentages of CD27+IgD+ and CD27+IgDneg memory B-cells were very low at birth and significantly increased only at 6 months (p=0.02 and p less than 0.001, respectively). We found a significant increase only in in vitro stimulated IgG production at 6 months as compared to birth (p less than 0.001). Moreover, a lower secretion of anti-Pn IgM antibodies up to 6 months of age, as compared to controls was observed. Our results underline that the susceptibility and severe course of infection in the neonate can be attributed, at least in part, to the lack of pre-existing immunological memory and competent adaptive immunity.

Non classical HLA genes and non-HLA genes in a population of infants at familial risk of atopy.

AIM: We investigated on parental history and IgE serum level in 2588 consecutive newborns to individuate babies "at risk" of atopy at birth and we analysed the polymorphisms of class III region to evaluate the association with immunogenetic markers of HLA: C4A, C4B, LTA, RAGE and TNFA genes; we performed TNF and IgE receptor (FCERB1) physiologically related gene polymorphisms. RESULT: 791 babies/2588 (30.6%) were considered "at risk" for atopy and followed-up: 400 had familial history of atopy (at least one parent or sibling), 256 had IgE >0.35 kUA/l at birth and during the follow-up and 135 were positive for both conditions. The allele C4B2 was significantly more frequent in the sample of babies at risk (22.1% vs 10%, p< 0.001). Furthermore, the mean value of IgE at birth in babies carrying the allele C4B2 was 2.26 KUA/l versus 0.74 KUA/l in those not carrying this allele (p=0.01). No significant association emerged for RAGE at the centromeric end of class III region and for LTA, TNFA at the telomeric one. TNFRI, TNFRII and FCERB1 gene polymorphisms also seemed not implicated. CONCLUSION: Our study confirms that HLA class III region seems involved in familial predisposition to atopy, and C4B gene probably acts as a marker of a more restricted subregion.

Factors associated with climacteric symptoms in women around menopause attending menopause clinics in Italy.

OBJECTIVE: To obtain data on correlates of climacteric symptoms in women around menopause attending menopause clinics in Italy. METHODS: Since 1997 a large cross sectional study has been conducted on the characteristics of women around menopause attending a network of first level menopause outpatient's clinics in Italy. A total of 66,501 (mean age 54.4 years) women are considered in the present paper. RESULTS: The odds ratios of moderate and severe hot flashes/night sweats were lower in more educated women and (for severe symptoms only) in women reporting regular physical activity. Depression, difficulty to sleep, forgetfulness and irritability tended to be less frequent in more educated women and (depression only) in women reporting regular physical activity. Parous women reported more frequently these symptoms. CONCLUSIONS: This large study confirms in Southern European population that low education, body mass index and low physical activity are associated with climacteric symptoms. Parous women are at greater risk of psychological symptoms.

Risk factors for type 2 diabetes in women attending menopause clinics in Italy: a cross-sectional study.

OBJECTIVE: To analyze risk factors for type 2 diabetes among women attending menopause clinics in Italy for counselling about the menopause. SUBJECTS: Women attending a network of first-level outpatient menopause clinics in Italy for general counselling about menopause or treatment of menopausal symptoms. METHODS: Cross-sectional study with no exclusion criteria. Type 2 diabetes was defined according to National Diabetes Data Groups Indications and the fasting blood glucose at an oral glucose tolerance test within the previous year. RESULTS: Out of the 44 694 considered in this analysis, 808 had a diagnosis of diabetes type 2 (1.8%). In comparison with women aged < 50 years, the multivariate odds ratios (OR) of type 2 diabetes were 1.31 (95% confidence interval (CI), 0.99-1.74) for women aged 50-52 years, 1.66 (95% CI, 1.27-2.17) at 53-56 years and 2.84 (95% CI, 2.20-3.67) in women aged > or = 57 years. Type 2 diabetes was less frequently reported in more educated women (OR high school/university vs. primary school = 0.44 (95% CI, 0.36-0.55)). Being overweight was associated with an increased risk of type 2 diabetes. In comparison with women reporting a low level of physical activity, the multivariate OR of type 2 diabetes was 0.67 (95% CI, 0.54-0.84) for women reporting regular physical activity. In comparison with premenopausal women, the multivariate OR of type 2 diabetes was 1.38 (95% CI, 1.03-1.84) in women with natural menopause. This finding was present also after allowing for the potential confounding effect of age. The multivariate OR of diabetes for users of hormonal replacement therapy was 0.58 (95% CI, 0.46-0.73). CONCLUSIONS: This large cross-sectional study suggests that postmenopausal women are at higher risk of type 2 diabetes after allowance for the effect of age. Other main determinants of risk of type 2 diabetes in women around menopause were low socioeconomic status and being overweight. Diabetes was found less frequently in those taking hormone replacement therapy.

Antigen-specific T cell response in infants after recombinant hepatitis B virus vaccination at birth: evaluation of T helper lymphocyte diversity.

Recombinant hepatitis B virus antigen (rHBsAg)-specific CD4+ T cell clones (TCC) were isolated and expanded from the peripheral blood of nine children vaccinated at birth against the hepatitis B (HB) virus. Four of them responded with protective antibody production (responders), three subjects were unable to produce detectable antibody levels even after revaccination (nonresponders), and two infants produced antibodies only after revaccination (slow responders). TCC were then characterized for their ability to produce cytokines known to be important for T cell expansion (interleukin-2, IL-2) and/or effector functions (IL-4, IFN-gamma, IL-10). Results demonstrated that the frequency of rHBsAg-specific TCC in the samples of nonresponders was comparable to or higher than that in the samples of responders. Nevertheless, the majority of TCC obtained from responders or from slow responders before revaccination displayed the T helper 1 (T(H1))-dominant phenotype, while the majority of TCC obtained from nonresponders were nonpolarized T lymphocytes. After revaccination, the distribution of the different T(H) subsets in slow responders was heterogeneous. Overall, our present data suggest that an absence or delay in developing an rHBsAg-specific antibody response to vaccination is not associated with the capacity to generate an Ag-specific T cell response. However, compared to responders, nonresponding infants react to the rHBsAg vaccination with a reduced capacity to expand and differentiate toward polarized T(H) cells.

Decline of maternal hepatitis A virus antibody levels in infants.

UNLABELLED: Hepatitis A is a common viral infection causing substantial morbidity and mortality. The anti-hepatitis A virus (HAV) vaccination in infants would guarantee control of the infection. However, the immunogenicity of the HAV vaccine in infants could be impaired by the presence of passively acquired maternal HAV antibodies. This study evaluated the prevalence of HAV antibodies in 103 women at delivery and in their babies in the first year of life. Eighteen mothers (17.5%) had anti-HAV serum level >10 mIU ml(-1). In their infants the anti-HAV level was still positive in 11 out of 18 (61.1%) at 12 mo. Two out of 85 infants born to anti-HAV-negative mothers and anti-HAV negative at birth were found to be positive at 5 mo of age. CONCLUSION: It is proposed that all women be screened at delivery for anti-HAV antibodies. Children born to anti-HAV-negative mothers could be vaccinated early during the first year of life, whereas vaccination could be postponed in children born to anti-HAV-positive mothers, if necessary.

Immediate and delayed two-stage post-mastectomy breast reconstruction with implants. Our experience of general surgeons.

BACKGROUND: Breast reconstruction after a mastectomy, today is considered the best choice to improve the quality of life for the patient. The aim of the procedure is to consider the aesthetic results obtained and the various involvements, comparing them with those already reported in literature. METHODS. There is no certain data of the number of breast reconstructions performed in Italy in the few years. In 1999, in Veneto, reconstruction surgery after total breast removal, represented only 16% of the operations. In our surgery Division, from 1996 to 2000, there were breast reconstructions performed, either immediately or delayed on 87 patients out of 118 after mastectomy. 73.7% none over the age of 60 (range 27-60). The breast removal technique used is the Madden, while, for the reconstruction we have put into effect the submuscle prosthetic implant; 63 patients were subject to follow-up treatment during the period of 10 to 36 months. RESULTS: Seventy-eight percent of the women have judged the final aesthetic results the same or better than expected. Based on reconstruction with prosthesis, is the preferred procedure, even with serious complications which do not always influence the final results. This preference is because of the reduced operation time and the psychological and physical benefits due to immediate restoral of the mammary volume and shape. CONCLUSIONS: Some problems (Garavaglia Law) have slowed down the diffusion in Italy of this type of treatment which is now receiving more and more request and approval by the women affected with breast cancer.

Family study of non-responsiveness to hepatitis B vaccine confirms the importance of HLA class III C4A locus.

Non-responsiveness to hepatitis B virus (HBV) vaccine in adults is strongly associated with HLA-C4AQ0,DRB1*0301,DQB1*02 haplotype. This association was also demonstrated in neonates who failed to mount a humoral response to challenge with HBV vaccine. About 4% of vaccinated newborns do not reach a protective antibody level (> or =10 mIU/ml) at seroconversion and 0.4% is a non-responder even after receiving a fourth dose of vaccine (true non-responders (TNR)); while 3.6% achieved an antibody level > or =10 mIU/ml (slow responders (SR)) only when reboostered with the fourth dose. In the present study we extend the vaccination and HLA typing to 91 family members of probands to understand better the possible parent-to-child transmission of this trait. A transmission disequilibrium test (TDT), performed in 27 families, showed that the C4AQ0 allele was almost always transmitted to probands, both TNRs and SRs. Although not statistically significant, the highest LOD score was obtained with C4A locus: 1.58. These results suggest the presence of a region regulating immune response against HBV vaccination near to or coincident with the C4A locus.

[Spontaneous ectopic contralateral pregnancy with unicornuate uterus. A case report]. / Gravidanza ectopica spontanea controlaterale ad utero unicorne. Caso clinico.

A case is reported of spontaneous ectopic pregnancy in a non patent fallopian tube of a patient with a contralateral unicornuate uterus without rudimentary horn. A hemoperitoneum, due to the rupture of the tube, occurred with a HCG level of 48 I.U./l. A laparoscopy was therefore performed and the fallopian tube removed.

Increment of recombinant hepatitis B surface antigen-specific T-cell precursors after revaccination of slow responder children.

The aim of the study was to investigate the in vitro T-cell response to recombinant hepatitis B (rHBsAg) in a group of children (defined as "slow responders") vaccinated at birth, presenting antibody levels < 10 mIU/ml after the vaccination schedule, and developing anti-rHBs antibodies after revaccination. T-cell mediated immune response towards rHBsAg was evaluated in 35 healthy children in "bulk" culture experiments (19 responders and 16 slow responders) and by limiting dilution analysis (nine responders and five slow responders) to quantify the frequency of proliferating T lymphocyte-precursors (PTL-p). Before the booster dose, lymphocytes from slow responder children failed to proliferate to rHBsAg, while a normal proliferation was observed in all responders. A statistically significant difference in rHBsAg-specific PTLp frequencies was observed between the two groups. Among the slow responder group, a significant increase of PTLp was observed after the supplementary vaccine dose.Nevertheless, PTLp frequencies remained significantly lower than those measured in responders. These results suggest a role for follow-up of slow responder children over time, in order to perform booster vaccination when inadequate anti-HBs titre is present.

[Intraoperative complications of 697 consecutive operative hysteroscopies]. / Complicanze intraoperatorie di 697 isteroscopie operative consecutive.

BACKGROUND: Complications due to hysteroscopy are relatively rare events. They occur more frequently with operative hysteroscopy than with diagnostic hysteroscopy. Exact complications rates are difficult to determine owing to the natural tendency to report successes but not complications. Recognition of these situations will lead to prevention; in fact, all the most serious complications of operative hysteroscopy can be avoided when proper precautions are taken and close communication is maintained among gynecologic surgeon, the anesthesiologist and nursing staff. The more clinically significant complications are: uterine perforation, haemorrhage and electrolyte imbalance. METHODS: Between January 1993 and December 1998, 697 women underwent operative hysteroscopy in our Department. Operative hysteroscopy was performed with continuous flow, high frequency resectoscope. Under general anesthesia the cervix was dilated to 10 mm and the uterine cavity was distended with 1.5% glycine solution or mannitol under 80 to 120 mmHg pressure. Resection with electrocoagulation was completed. The patients were submitted to the following procedures: 354 endometrial polypectomies (50.7%), 160 myomectomies (23%), 114 endometrial ablations (16.4%) and 69 hysteroscopic metroplasties (9.9%). RESULTS: In our series complications occurred in 95 out of 697 patients (13.6%). The most important complications were: 12 (1.7%) uterine perforations, 48 (6.9%) intraoperative haemorrhages and 35 (5%) excessive hypotonic fluid absorptions. Four out of 12 perforations occurred during the dilation of the cervical channel. Since the distention of the uterine cavity could not be achieved, the procedures were stopped. No signs of vaginal or intraperitoneal haemorrhage were observed; 8 out of 12 perforations were due to the tip of the electrical source. The operative hysteroscopies were immediately stopped and the consequences were: 6 diagnostic laparoscopies, 1 laparotic hysterectomy (hemorrhage) and 1 laparotomy for thermal bowel injury. In 48 patients intraoperative bleeding could not be controlled with electrocautery. In these cases in the operating room a Foley catheter was inserted into the uterine cavity and the bulb inflated with 10 to 30 mL of liquid to tamponade the bleeding. The catheters were removed 12 to 24 hours later. No patients required blood transfusion. Excessive intravasation of electrolyte-free fluid occurred in 35 patients. Hyponatremia and hypokalemia (hypo-osmolarity result) were never serious. Headaches, nausea and vomiting were the most frequent symptoms of our patients. No cardiac arrhythmia, cerebral edema, brain herniation occurred. In our series, hemorrhage was the most common complication; intravasation and uterine perforation were at the second and the third place. Complications rates decreased progressively du to a better major training and experience of the surgeons. Also the curves of each complication show a significant decrease. Myomectomy in our hands has been the most dangerous procedure. However, serious sequelae were rare mainly for two reasons: we prefer stop the intervention rather than continue when a deficit of 1.000 mL is reached. Consequently, it is very important to discuss the possibility of incomplete resection of the endouterine lesion with the patient preoperatively; a protocol for fluid management in the operating room must be used for all the procedures (also the easiest) by all the surgeons and the nurses. CONCLUSIONS: Our relatively high prevalence of intraoperative complications and distribution of the different types do not differ from the findings of published reports. In personal experience operative hysteroscopy is a safe surgical procedure for the treatment of endouterine abnormalities.

Insulin receptor antibodies inhibit insulin uptake by the liver: in vivo 123I-insulin scintigraphic scanning and in vitro characterization in autoimmune hypoglycemia.

BACKGROUND: Insulin receptor antibodies can induce severe hypoglycemia or insulin resistance in rare autoimmune syndromes. In vitro properties of these antibodies occasionally explain the clinical features of the syndrome, but direct evidence of their in vivo activity is poor. We studied a 58-year-old male with rheumatoid arthritis who presented with hypoglycemic coma. METHODS AND RESULTS: Antibodies were detected by inhibition of 125I-insulin binding to human insulin receptor-3T3 cells by the patient's serum. By immunofluorescence, they were immunoglobulin G of all four subclasses, immunoprecipitated insulin receptors from biotin-labeled cells, and triggered phosphorylation of the beta subunit of the insulin receptor. Insulin binding on the patient's red blood cells was markedly reduced. A biodistribution study after intravenous 123I-Tyr A14 insulin showed a marked inhibition of tracer uptake by the liver, reaching 10% of the injected dose (controls, mean +/- SD, 21.1 +/- 1.7%; n = 10). Time activity curves generated on the liver and on the heart were parallel, with a T1/2 of 11.5 minutes for both, suggesting that no specific uptake occurred in the liver, where tracer activity represented only the blood pool. Clearance of insulin from the blood was indeed slower than in controls and mainly occurred through the kidneys. Analysis of plasma 123I-insulin immunoreactivity and trichloroacetic acid precipitate showed that insulin degradation did not occur as in normal controls. CONCLUSIONS: In this patient with hypoglycemic syndrome, insulin receptor antibodies with in vitro insulin-like activity are capable of blocking in vivo the access of insulin to the liver receptor compartment, as directly demonstrated by the markedly altered biodistribution of intravenously injected 123I-insulin.

Humoral response to recombinant hepatitis B virus vaccine at birth: role of HLA and beyond.

From 1991 to 1998 we vaccinated 4835 neonates against hepatitis B virus (HBV) and monitored their humoral response to the recombinant vaccine. In a sample of 184 of these babies we studied the association between HLA class I and II genomic polymorphisms and humoral response to the vaccine and the association between the response and immune-mediated diseases. A subgroup of 96 babies also underwent HLA class III (C4A and C4B) typing. Four levels of humoral response were identified, each with a peculiar MHC restriction. Different HLA products seem to act as agonists (C4AQ0 and HLA-DQB1(*)02) or antagonists (C4AQ0, HLA-DQB1(*)02, and HLA-DRB1(*)11, DQB1(*)0301) in lowering humoral response to HBV vaccine. The group of responders was characterized more for lacking "nonresponder" alleles than for having specific "responder" ones. Tolerance to HBV peptides may have clinical implications, possibly being a marker for babies with a genetic risk of immunopathologies. In fact, many of the poor responders carried from two to four HLA-DQ alpha beta heterodimers predisposing to insulin-dependent diabetes mellitus and celiac disease. Two true nonresponders suffered from allergies and two slow responders had transient episodes of hyperglycemia.

[Endometrial carcinoma: prognostic significance of cellular ploidy]. / Carcinoma endometriale: significato prognostico della ploidia cellulare.

BACKGROUND: The aim of the paper is to compare ploidy status with the traditional prognostic factors (grading, myometrial invasion, cytology of peritoneal fluid and node invasion) in 118 women affected by endometrial carcinoma and treated by hysterectomy in our department in order to evaluate a relationship between these parameters. METHODS: Since January 1988 and August 1996 127 women (average age: 61) affected by endometrial carcinoma and not previously treated have been submitted to abdominal or vaginal hysterectomy. A retrospective study was carried out on 118 of these women evaluating DNA ploidy on fixed neoplastic samples through flow cytometry (Coulter Elite with Argon Laser). Aneuploidy was defined as cell population containing at the same time two or more moderate peaks in G0/G1. Histology of neoplastic tissues could evaluate grading and myometrial invasion in all cases. Cytology of peritoneal fluids and nodal state were evaluated respectively in 99 and 56 patients. The results obtained have been compared by Fisher's statistical test. RESULTS: 70.3% of evaluated neoplasias were diploid, while 29.7% were aneuploid. No statistical difference was observed comparing ploidy status with every considered parameter. CONCLUSIONS: The results obtained show that DNA ploidy doesn't seem to be positively correlated with any traditional histopathological factors. The literature about this matter is questionable. Histopathological analysis is the only prognostic factor and it is the only parameter to personalize treatment.

Circulating GH isoforms and GH bioactivity in preterm neonates.

Among the molecular variants of human GH, the monomeric 22-kD is the predominant isoform, whereas the 20-kD is the second most abundant isoform. Because little is known on the pattern of human GH isoforms in the early postnatal period, we evaluated serum levels of 22-kD GH by an immunofluorometric assay and of 20-kD GH by an ELISA using an anti-20-kD antibody, and measured GH bioactivity with the Nb2 cell bioassay in 19 preterm neonates (gestational age, 32 +/- 0.5 wk; mean +/- SEM) on the fourth and 15th days of life. As control subjects, we studied 19 full-term neonates (gestational age, 39 +/- 0.3 wk) on the fourth day of life and 20 healthy adults, aged 20 +/- 0.3 y. Four-day-old preterm neonates showed significantly higher serum values of 20-kD GH (0.99 - 0.14 ng/mL) than full-term neonates (0.33 +/- 0.07 ng/mL; p < 0.001) and adults (0.09 +/- 0.02; p < 0.0001). Likewise, 22-kD GH and GH levels by Nb2 cell bioassay were also significantly higher (p < 0.001) in preterm than in full-term neonates and young adults. A significant decrease (p < 0.01) in 20-kD, 22-kD, and Nb2-determined GH was observed in preterm neonates on the 15th day of life The percentage of the 20-kD isoform was similar in the preterm infants at the fourth and 15th day, in full-term-infants, and in adults (2.7%, 2.7%, 2.8%, and 3.16%, respectively). Our results indicate that 20-kD GH serum levels change throughout life as regards total amount, but not as regards percentage.

Vertical transmission of hepatitis C virus and follow-up of newborns from infected mothers.

BACKGROUND: Aim of the study is to analyze the rate of vertical transmission of HCV and the time of clearance of maternal antibodies in non-infected babies serum. METHODS: We have studied 36 babies born to HCV-positive and HIV-negative pregnant women at the University of Pavia. All mothers underwent blood tests to evaluate the presence of anti-HCV antibodies and viral RNA during pregnancy and after delivery. All babies underwent several tests at different times to evaluate the presence of viral RNA and the clearance of maternal antibodies. RESULTS: All babies proved HCV-Ab positive at birth, but only one case (2.7%) proved infected at PCR analysis. Different patterns of HCV-Ab clearance were noted in the 35 non-infected babies. Of 24 babies from HCV-RNA-positive mothers, HCV-Ab reached zero in 24 months while in 11 babies from HCV-RNA-negative mothers, the antibodies disappeared at 12 months. A statistical difference was noted between the two groups of babies for the time of clearance of antibodies. CONCLUSIONS: The risk of vertical transmission in babies born to HCV-RNA negative mothers is very low, and the clearance of maternal antibodies is set within 12 months of follow-up. Mothers positive to HCV-RNA have a higher risk of transmitting the virus to their offspring and the time of clearance of antibodies in non-infected babies seems to be longer. A correct follow-up of these children must be no shorter than 24 months.

Early immunisation with hepatitis B vaccine: a five-year study.

We evaluated the response to the recombinant Hepatitis B vaccine using an accelerated schedule versus the traditional schedule by studying the immunologic memory induced in 200 children with HBs-Ag negative mothers. At seroconversion, the traditional schedule presented a higher percentage of children with serum HBs-Ag concentrations over 100 mIU/ml than the accelerated schedule. After five years this difference was no longer statistically significant and children who presented anti-HBsAg concentrations below 10 mUI/ml received an additional booster dose which stimulated the antibody concentration to exceed 100 mIU/ml in all cases. Recombinant HBV vaccine induced better long term immunologic memory when it was administered earlier.