Susana Vighi

Apoptosis and expression of Bcl-2 and Bax in eutopic endometrium from women with endometriosis

OBJECTIVE To evaluate and compare spontaneous apoptosis and Bcl-2 and Bax expression in eutopic endometrium from women with and without endometriosis. DESIGN Apoptosis and Bcl-2 and Bax expression were examined in eutopic endometrium from women with and without endometriosis. SETTING Instituto de Biología y Medicina Experimental-CONICET, Department of Gynecology and Department of Gynecological Pathology, Clínicas University Hospital, Buenos Aires, Argentina. PATIENT(S) Women with untreated endometriosis (n = 14) and controls (n = 16). INTERVENTION(S) Collection of endometrial samples during diagnostic or therapeutic laparoscopy. MAIN OUTCOME MEASURE(S) Apoptotic cells were detected with use of the dUTP nick-end labeling (TUNEL) assay; Bcl-2 and Bax expressions were assessed with use of immunohistochemical techniques. RESULT(S) Spontaneous apoptosis was significantly lower in eutopic endometrium from patients with endometriosis, compared with healthy controls (2.26 +/- 0.53 and 9.37 +/- 1.69 apoptotic cells/field, respectively) and was independent of cycle phase. An increased expression of Bcl-2 protein was found in proliferative eutopic endometrium from patients with endometriosis. Bax expression was absent in proliferative endometrium, whereas there was an increase in its expression in secretory endometrium from both patients and controls. CONCLUSION(S) Women with endometriosis show decreased number of apoptotic cells in eutopic endometrium. The abnormal survival of endometrial cells may result in their continuing growth into ectopic locations.

Lymph node metastases in carcinoma of the cervix uteri: Response to neoadjuvant chemotherapy and its impact on survival

One hundred and sixty-nine patients with squamous cancer of the cervix uteri treated with three courses of neoadjuvant chemotherapy with the modified VBP scheme are presented. All were subjected to a Wertheim-Meigs operation with paraaortic lymph-adenectomy. The incidence of lymph node metastases is analyzed according to clinical stage, tumor volume, residual tumor in the surgical specimen, and clinical response to neoadjuvant chemotherapy. A significant decrease in the incidence of lymph node involvement was observed in good responders. Survival rates, after 2 years of follow-up, improved in those cases with small residual tumor, negative parametria, and negative nodes.

Results of a Phase II trial with neoadjuvant chemotherapy in carcinoma of the cervix uteri

Results of a Phase II trial with neoadjuvant chemotherapy in carcinoma of the cervix uteri (VBP modified scheme) show that 85.7% of patients given this therapy were NED in Stage IIb versus 54% of a nonrandomized control group given conventional therapy. In Stage IIIb the averages are 66.6% vs. 31% in the control group. Analysis of the ecographic data has shown that if a critical prechemotherapy volume (120 cm3) is exceeded, the prognosis is unfavorable, especially in cases treated with radiotherapy as second-line treatment.

Prenatal Hyperandrogenization Induces Metabolic and Endocrine Alterations Which Depend on the Levels of Testosterone Exposure

Prenatal hyperandrogenism is able to induce polycystic ovary syndrome (PCOS) in rats. The aim of the present study was to establish if the levels of prenatal testosterone may determine the extent of metabolic and endocrine alterations during the adult life. Pregnant Sprague Dawley rats were prenatally injected with either 2 or 5 mg free testosterone (groups T2 and T5 respectively) from day 16 to day 19 day of gestation. Female offspring from T2 and T5 displayed different phenotype of PCOS during adult life. Offspring from T2 showed hyperandrogenism, ovarian cysts and ovulatory cycles whereas those from T5 displayed hyperandrogenism, ovarian cysts and anovulatory cycles. Both group showed increased circulating glucose levels after the intraperitoneal glucose tolerance test (IPGTT; an evaluation of insulin resistance). IPGTT was higher in T5 rats and directly correlated with body weight at prepubertal age. However, the decrease in the body weight at prepubertal age was compensated during adult life. Although both groups showed enhanced ovarian steroidogenesis, it appears that the molecular mechanisms involved were different. The higher dose of testosterone enhanced the expression of both the protein that regulates cholesterol availability (the steroidogenic acute regulatory protein (StAR)) and the protein expression of the transcriptional factor: peroxisome proliferator-activated receptor gamma (PPAR gamma). Prenatal hyperandrogenization induced an anti-oxidant response that prevented a possible pro-oxidant status. The higher dose of testosterone induced a pro-inflammatory state in ovarian tissue mediated by increased levels of prostaglandin E (PG) and the protein expression of cyclooxygenase 2 (COX2, the limiting enzyme of PGs synthesis). In summary, our data show that the levels of testosterone prenatally injected modulate the uterine environment and that this, in turn, would be responsible for the endocrine and metabolic abnormalities and the phenotype of PCOS during the adult life.

Cytosolic accumulation of HPV16 E7 oligomers supports different transformation routes for the prototypic viral oncoprotein: The amyloid–cancer connection

E7 is the major transforming activity in human papillomaviruses, a causal agent for cervical cancer. HPV16 E7 is a small protein with a natively unfolded domain for which dozens of specific cellular targets were described, and represents a prototypical oncoprotein among small DNA tumor viruses. The protein can form spherical oligomers with amyloid‐like properties and chaperone activity. Conformation specific antibodies locate endogenous oligomeric E7 species in the cytosol of 3 model cell lines, strongly co‐localizing with amyloid structures and dimeric E7 localizes to the nucleus. The cytosolic oligomeric E7 appear as the most abundant species in all cell systems tested. We show that nuclear E7 levels are replenished dynamically from the cytosolic pool and do not result from protein synthesis. Our results suggest that long‐term events related to de‐repression of E7 would cause accumulation of excess E7 into oligomeric species in the cytosol. These, together with the known target promiscuity of E7, may allow interactions with many of the non‐pRb dependent targets described. This hypothesis is further supported by the detection of E7 oligomers in the cytosol of cancerous cells from tissue biopsies.

Vulvar infection caused by human papilloma virus in children and adolescents without sexual contact

Abstract We studied the indirect transmission of human papilloma virus (HPV) to young children and adolescents who were proven, by in-depth sexual history and gynecological examination, never to have had sexual intercourse. Two groups were identified: group A, daughters and sisters (age less than 18 years) of 31 adults with HPV genital lesions (n = 40); group B, 10 virginal girls and adolescents with HPV vulvar lesions and the adult household members were studied for the HPV infection (n = 26). Evaluations included PAP smear, colposcopy, vaginoscopy, and vulvoscopy in the adult female; peneoscopy and biopsy in males; vulvoscopy and cytologic studies (labia minora, vaginal washing) in all minors. A biopsy was obtained from all lesions. Vulvoscopic findings consistent with HPV were found in 75% of the group A girls. In 83% of those, the biopsy was positive. In group 2, 90% were found to live in a household with at least one positive adult (previously undetected). One or more of the following practices were discovered in the majority of cases: occasional bed sharing, and joint use of towels, bathing suits, underwear, and tub bath. Postdefecation hygiene was carried out in the family bidet. This supports the hypothesis that HPV can be nonsexually transmitted.

Vulvar carcinoma in situ: A report of 28 cases

Abstract Twentyeight cases of vulvar CIS are reported. The interest of a group of specialists in this pathology notably improves early detection. Biopsy was the basic diagnostic method. The CIS vulvar patients are generally younger than those suffering invasive cancer. The association with other genital or extragenital neoplasias is analyzed. Seven cases were asymptomatic. Unifocal lesions were more frequent than multifocal lesions. Individualized treatment is considered.

Is subradical surgical treatment for carcinoma of the cervix uteri stage IB logical

Twenty-eight patients with squamous carcinoma of the cervix FIGO stage Ib were treated with three courses of neoadjuvant chemotherapy with a VBP modified scheme. Clinical responses showed that the percentage of complete and moderate responses exceeds 95% of the cases. Clinical response was also related to tumor bulk measurement by ultrasound scanning. Twenty-three of the patients were then subjected to the Wertheim-Meigs operation. Pathological findings of surgical specimens showed absence of residual lesion in 6 patients (26.1%) and carcinoma smaller than 0.5 cm in 5 patients (21.7%). Tumor response to neoadjuvant chemotherapy was excellent in NG3, MG3 tumors when lymphoplasmomonocytic infiltration was present. In accord with this result a new protocol was developed.

Apoptosis is increased and cell proliferation is decreased in out-of-phase endometria from infertile and recurrent abortion patients

BackgroundVarious endometrial abnormalities have been associated with luteal phase deficiency: a significant dyssynchrony in the maturation of the glandular epithelium and the stroma and a prevalence of out-of-phase endometrial biopsy specimens. Out-of phase endometrium is a controversial disorder related to failed implantation, infertility and early pregnancy loss. Given that the regulation of the apoptotic process in endometrium of luteal phase deficiency is still unknown, the aim of this study was to evaluate cell proliferation, apoptosis and the levels of the main effector caspase, caspase-3 in the luteal in-phase and out-of-phase endometrium.MethodsThirty-seven endometrial samples from sterile or recurrent abortion patients were included in this study: 21 in-phase samples (controls) and 16 samples with out-of-phase endometrium. Biopsy specimens of eutopic endometrium were obtained from all subjects during days 21-25 of the menstrual cycle. The endometrium with endometrial maturity of cycle day 25 or less at the time of menstruation was considered out-of phase. Endometrial tissues were fixed in 10% buffered formaldehyde. For apoptosis quantification, sections were processed for in situ immunohistochemical localization of nuclei exhibiting DNA fragmentation, by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP digoxygenin nick-end labeling (TUNEL) technique. Expressions of Proliferating Cell Nuclear Antigen (PCNA) as a marker of cell proliferation, and of cleaved caspase-3 as a marker of apoptosis, were assessed by immunohistochemistry in the luteal in-phase and out-of-phase endometrium from infertile and recurrent abortion patients.ResultsLuteal out-of-phase endometrium had increased apoptosis levels compared to in-phase endometrium (p < 0.05). Caspase-3 evaluation confirmed these results: the luteal out-of-phase endometrium showed augmented cleaved caspase-3 expression (p < 0.005). As well, our data demonstrated that the luteal out-of-phase endometrium expresses decreased PCNA levels (p < 0.05), showing that cell proliferation is diminished in this tissue.Conclusionsthis study represents the first report describing variations at the cell proliferation and cell death levels in the out-of-phase endometrium in comparison with in-phase endometrium from infertile and recurrent abortion patients. Further studies are needed to elucidate a potential role of these alterations in the physiopathology of luteal phase deficiency.

Detrimental effects of hyperandrogenism on uterine functions

The aim of the present work was to study some of the adverse effects produced by hyperandrogenism on the uterine function. Daily injection of dehydroepiandrosterone (DHEA: 6 mg/ 100 g body weight, sc) for 20 consecutive days induced polycystic ovaries in BALB/c mice. In this model, we found that DHEA produced alterations on uterine histology closely related to the development of tumour structures. In addition, hyperandrogenism induced a pro-inflammatory and a pro-oxidant condition represented by increased levels of prostaglandin F2 alpha production and uterine nitric oxide synthase (NOS) activity and by a decrease in both superoxide dismutase (SOD) and catalase (CAT) activities together with a decrease in the levels of the antioxidant metabolite glutathione (GSH). DHEA also induced an increase in CD4+ together with a decrease in the CD8+ T lymphocytes that infiltrate the uterine tissue. We conclude that this intricate network of regulators could be responsible for the low rate of implantation observed in women with polycystic ovary syndrome.