Maria Antonieta Longo Galvão

Histologic classification of specimens from women affected by superficial endometriosis, deeply infiltrating endometriosis, and ovarian endometriomas

In this retrospective observational study involving 176 patients and 271 biopsies, the histologic differentiation in superficial endometriosis, deeply infiltrating endometriosis, and ovarian endometriomas was evaluated according to a previously proposed classification system. Results showed a predominance of the undifferentiated glandular pattern (33.5%) and mixed glandular pattern (46.9%) in deeply infiltrating endometriosis lesions, whereas the well-differentiated glandular pattern (41.8%) was most frequently seen in superficial endometriosis lesions, and in ovarian endometriomas a predominance of both the undifferentiated (40.5%) and mixed patterns (37.8%) was observed.

Effects of Trifolium pratense and Cimicifuga racemosa on the endometrium of Wistar rats.

OBJECTIVE The aim of this study was to evaluate the effects of Trifolium pratense and Cimicifuga racemosa upon the endometrium of castrated female Wistar rats, comparing these results with a placebo and estradiol valerate. METHODS Thirty-two adult castrated female Wistar rats were divided into four groups (eight rats per group) that receiving either tap water, estradiol valerate, isoflavones from T. pratense or deoxyactein from C. racemosa daily. The doses used were equivalent to normal doses used in humans. The results were analyzed by endometrial histology and the expression of alpha-estrogen receptor and protein Ki67. Both alpha-receptor and Ki67 expressions were determined by immunohistochemical and morphometric analysis. RESULTS Endometrium histology stayed atrophic with both herbal extracts, but T. pratense supplementation increased the expression of alpha-estrogen receptors when compared to the placebo group, without protein Ki67 expression enhancement. Both herbal extracts presented a lower Ki67 expression when compared to the placebo group. CONCLUSION T. pratense presented alpha-estrogen receptor stimulation in the endometrium without increasing cell proliferation. Both herbal extracts reduced endometrial proliferation in comparison to the placebo group.

Evaluation of density of Langerhans cells in human cervical intraepithelial neoplasia

Objective. To compare the number of Langerhans cells in areas of CIN 3 and in areas without histopathological alteration adjacent to CIN 3 with those found in histologically normal control cases. Material and methods. Specimens were obtained from 46 normal cervixes from women undergoing hysterectomies due to uterine leiomyomata and 71 from conization specimens for CIN 3. The identification of the Langerhans cells was performed by immunohistochemical analysis, using antibodies to S‐100 protein. The number of intraepithelial Langerhans cells was counted at a 400× magnification under light microscope and a 10 field count was performed in areas of epithelium of the normal cervix (group A), areas of CIN 3 in the conization specimens (group B), and areas of epithelium without histopathologic alteration adjacent to CIN 3 (group C). Results were expressed as number of cells per square millimeter of epithelium. Results. Comparing groups A and B, we observed that the number of Langerhans cells was significantly higher in the latter group; between groups A and C, the number of cells was significantly lower in the second group. The number of Langerhans cells was also compared between groups B and C and it was significantly lower in the latter, independent of smoking habits. Conclusions. We theorize that the increase in the population of Langerhans cells in areas of CIN 3 could be explained by migration of these cells from adjacent areas without histopathologic alterations in order to act in the restraint of the development of neoplasia; cigarette smoking did not influence this migration.

Effects of Glycine max (L.) Merr. soy isoflavone vaginal gel on epithelium morphology and estrogen receptor expression in postmenopausal women: A 12-week, randomized, double-blind, placebo-controlled trial

OBJECTIVE Evaluate the effects of vaginal administration of isoflavones derived from Glycine max (L.) Merr. as a treatment option for vaginal atrophy, on the morphology and expression of estrogen receptors in vaginal epithelium of postmenopausal women. METHODS The double-blind, randomized, placebo-controlled, clinical trial. Sixty women were treated for 12 weeks with isoflavone vaginal gel 4% (1g/day) and a placebo gel. After 4 and 12 weeks, the vaginal atrophy symptoms were classified at none, mild, moderate and severe and the vaginal cytology were taken to determine the maturation value. Vaginal pH was measured at the beginning and end of therapy. Microbiopsies in vaginal fornix were performed before the treatment and after 12 weeks of treatment. RESULTS Isoflavone vaginal gel was effective for relief of vaginal dryness and dyspareunia symptons and an increase in the intermediate and superficial cells was noted. The vaginal pH in the isoflavone group was 7.1 at baseline and 5.4 after 12 weeks, whereas in the placebo group there was no significant change. A significant increase in thickness after treatment was detected in the Isoflavone Group. The percentage of estrogen receptor positive cells in vaginal epithelium for the Isoflavone Group ranged from 58.5% at the beginning of treatment to 82.6% after 12 weeks. These results were superior to placebo gel. CONCLUSION Glycine max (L.) Merr. at 4% vaginal gel on a daily basis in postmenopausal women led to improvements in vaginal atrophy symptoms, maturation values, vaginal pH, morphology and expression of estrogen receptors in vaginal epithelium. Isoflavones proved good treatment options for relief of vulvovaginal atrophy.

The effect of cigarette smoking on cervical langerhans cells and T and B lymphocytes in normal uterine cervix epithelium.

The objective of our study was to evaluate the effect of smoking on intraepithelial Langerhans cells and T and B lymphocytes in normal cervical epithelium. A total of 47 women who underwent hysterectomies because of uterine leiomyomata were investigated. Thirty-two individuals were nonsmokers and 15 were current smokers. A segment of tissue was taken from the cervix after the removal of the uterus and was formalin-fixed and paraffin-embedded for histologic analysis. The identification of the Langerhans cells and T and B lymphocytes was carried out by immunohistochemical analysis. The number of intraepithelial Langerhans cells was expressed by the amount of cells per square millimeter of epithelium. For T and B lymphocytes, a score expressed the cell count. The comparison of the number of intraepithelial Langerhans cells between smoking and nonsmoking women showed a significant difference (P=0.045), but it did not occur in relation to the number of T and B cells between the 2 groups. There was also no significant difference in relation to the number of cigarettes smoked per day, time of consumption, and total amount of cigarettes smoked throughout the lifetime. It was shown that smoking reduces the number of intraepithelial Langerhans cells in a normal cervix but does not influence the number of T and B lymphocytes.

Application of an easy and useful morphometric technique for immunohistochemistry counting

We have been performing several scientific works related to prognostic factors of preneoplasic and neoplasic lesions of the cervix, due to the presence of post-graduation students in our department. In order to perform such studies we have been using immunohistochemistry with a certain frequency. When working with immunohistochemistry, sometimes we wish to determine the concentration of a certain cell that is being analyzed in order to compare them in different situations. Such cell count, according to a certain space unit, is called morphometry. The term morphometry is comprised by the Greek radical — morphé, which means shape, related to the Greek radical — metrikós, or to the Latin — metricu, which means act of measuring or process of establishing measurements. Thus, several computer softwares of morphometry were developed and are commercially available to use in daily clinical practice. However, in centers with scarce resources, as in our case, purchasing such softwares is not viable due to their high cost. Thus, instead of using these softwares, one of the most common ways of getting such an estimate would be through counting cells in a regular optical microscope. Due to that setback in getting such morphometry softwares, we have developed a morphometric method that is quite easy to perform, and that can be used by any research center, by making use of a Neubauer chamber, also known as Hemocytometer or Counting Chamber, which is easily found in clinical pathology labs. The Neubauer Chamber consists of a glass slide, thicker than a regular slide, where there are usually two chambers engraved in the glass (two darker parts in the center). Each Neubauer Chamber has the design of a grid, dividing it into squares with known dimensions, which are visualized only through the optical microscope. The Neubauer Counting Chamber is divided into 9 large squares of 1 mm each side by means of triple lines (the area of a large square=1 mm). Each large square contains 16 (4×4) medium size squares. Eachmedium size square is further divided by triple lines into 25 (5×5) small squares, eachwith side 1/5 of a mm. Thus the dimension of an individual small square of 0.05×0.05 mm=0.0025 mm and 0.1 mm deep (Fig. 1). The system we implemented used a regular optical microscope adapted to a microcamera and screen. Thus, the image observed through the microscope lenses is captured by the video camera and showed in the chosen monitor. The Neubauer Chamber is then placed in the optical microscope, under a objective which increases 400 times the original size of the object (large magnification field), or any other increase chosen by the researcher. At last, the area of the large magnification field (400× magnification) is estimated, in mm, by counting the total number of individual small squares of 0.0025 mm of the chamber present in the magnification area that was chosen. In our evaluation, the total area of a large magnification corresponded to 0.094 mm. In such a manner, each section prepared by immunohistochemical method was evaluated under light microscope. The number of cells in study was counted at 400× magnification. A 10 field count was performed in the different situations we were evaluating. The mean number of cells present in this 10 field count represents, as mentioned before, 0.094 mm. By means of a simple rule of three, the number of cells is converted into 1 mm. The cells count was expressed as numerical densities, i.e. number of positive cells per square millimeter of epithelium. Available online at www.sciencedirect.com

Células glandulares atípicas em esfregaços cervicovaginais: significância e aspectos atuais

Atypical glandular cells (AGC) on cervical smears are unusual but important cytologic diagnosis. The incidence of AGC ranges from 0.08% to 0.81 % of all cervical smears tests. The 2001 Bethesda System nomenclature classifi es these glandular lesions as AGC not otherwise specifi ed, AGC favor neoplasia, endocervical adenocarcinoma in situ (AIS) and invasive adenocarcinoma. Of women with AGC smears, a great number will have no histologic abnormalities on further evaluation. However, 17.4% to 62.2% are found to have signifi cant histologic lesions such as cervical intraepithelial neoplasia, AIS, squamous and glandular cancers from sites farther the cervix and endometrium. The risk of signifi cant disease is related to the AGC subclassifi cation that was found. Because of the high likelihood that AGC is associated with signifi cant histological disease, simply repeating the cytological smear is not suffi cient for the management of this condition. This review describes the epidemiology, evaluation and management of patients with these citologic readings.Atypical glandular cells (AGC) on cervical smears are unusual but important cytologic diagnosis. The incidence of AGC ranges from 0.08% to 0.81 % of all cervical smears tests. The 2001 Bethesda System nomenclature classifies these glandular lesions as AGC not otherwise specified, AGC favor neoplasia, endocervical adenocarcinoma in situ (AIS) and invasive adenocarcinoma. Of women with AGC smears, a great number will have no histologic abnormalities on further evaluation. However, 17.4% to 62.2% are found to have significant histologic lesions such as cervical intraepithelial neoplasia, AIS, squamous and glandular cancers from sites farther the cervix and endometrium. The risk of significant disease is related to the AGC subclassification that was found. Because of the high likelihood that AGC is associated with significant histological disease, simply repeating the cytological smear is not sufficient for the management of this condition. This review describes the epidemiology, evaluation and management of patients with these citologic readings.

Classificação histológica e qualidade de vida em mulheres portadoras de endometriose

PURPOSE To assess the relationship between the histological classification and the quality of life of patients operated for endometriosis. METHODS A cross-sectional observational study, with assessment of 32 biopsies of the intestine, peritoneum and uterosacral ligament from 40 women with deep endometriosis. The quality of life (QOL) was determined by applying the SF-36 questionnaire pre-operatively and at 6 and 12 months postoperatively. Biopsies were histologically classified into pure stromal (EP), glandular differentiated (GD), glandular undifferentiated (GI) and mixed (GM), remaining in the sample only GI and GM, which are related to eight domains of the SF-36. RESULTS According to the histologic type, the following distribution was observed: peritoneum 63% GI and 35% GM; intestine 19% GI and 24% GM; uterosacral ligament with 41% GI and 35% GM. Regarding the QOL and the histological classification, in the intestine only GM was associated with improvement of social and emotional aspects from 0 to 6 months; the domains general health status (p=0.01) and social aspect (p=0.04) were significantly related to improvement of the QOL from 0 to 6 months, and the general health status tended to improve from 0 to 12 months. Regarding pain (p=0.06) and the emotional aspect (p=0.05), the QOL tended to improve from 0 to 6 months and the vital capacity (p=0.1) improved from 0 to 6 months and from 0 to 12 months. Regarding the emotional aspect, evolution of the two histological types was not favorable for improvement in MG from 0 to 6 months. No significant relationships between histologic type and QOL were evident in the uterosacral ligament samples. CONCLUSION Improvement in the QOL of women undergoing laparoscopic surgery for deep endometriosis is associated with the histologic grade. The peritoneal biopsy of GI revealed improved QOL after surgery.

Cyclooxygenase-2 and human epidermal growth factor receptor type 2 (HER-2) expression simultaneously in invasive and in situ breast ductal carcinoma

CONTEXT AND OBJECTIVE Cyclooxygenase-2 (COX-2) and human epidermal growth factor receptor type 2 (HER-2) are associated with tumorigenesis. Studies have shown that HER-2 can regulate COX-2 expression. The aim of this study was to evaluate the correlation between COX-2 and HER-2 expression in normal breast epithelium and in ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) present in the same breast. DESIGN AND SETTING Cross-sectional study at the Mastology Unit of the Department of Gynecology and Obstetrics, Santa Casa de Misericórdia de São Paulo Hospital. METHODS COX-2 and HER-2 were detected using immunohistochemistry on 100 tissue fragments. HER-2 > +2 was subjected to fluorescence in situ hybridization (FISH). RESULTS COX-2 expression was detected in 87%, 85% and 75% of IDC, DCIS and normal epithelium, respectively. HER-2 expression was detected in 34% of IDC and 34% of DCIS. COX-2 in DCIS correlated with HER-2 in IDC (P = 0.049) and DCIS (P = 0.049). COX-2 in normal epithelium correlated with HER-2 in IDC (P = 0.046) and DCIS (P = 0.046). COX-2 in IDC was not associated with HER-2 (P = 0.235). Comparison between COX-2 and HER-2 in DCIS showed that there was a statistically significant difference with regard to nuclear grades II and III and presence of comedonecrosis (P < 0.001). In IDC, there was significant expression with nuclear grades II and III and histological grade II (P < 0.001). CONCLUSIONS Our findings provide evidence that HER-2 and COX-2 regulate each other.

Avaliação histológica de pólipos endometriais em mulheres após a menopausa e correlação com o risco de malignização

OBJETIVOS: caracterizar histologicamente polipos endometriais em mulheres apos a menopausa e avaliar o risco de lesoes hiperplasicas e neoplasicas neles contidos. METODOS: analise retrospectiva de 82 casos de mulheres portadoras de histopatologia confirmatoria de polipo endometrial, submetidas a polipectomia histeroscopica devido ao diagnostico de espessamento endometrial a ultra-sonografia transvaginal. Todos os prontuarios pertinentes foram revisados para obtencao da historia clinica, antecedentes, dados referentes ao exame histeroscopico e resultado final da histopatologia. RESULTADOS: das 82 mulheres avaliadas, 10,9% faziam uso de algum tipo de terapia hormonal e 28 (34,1%) queixavam-se de sangramento por via vaginal. Polipo solitario foi encontrado em 56 mulheres (68,3%), em 19 casos (23,2%) foram encontrados dois polipos e em 7 casos (3,6%), tres ou mais polipos. A avaliacao histologica definitiva evidenciou polipo benigno em 63 mulheres (76,8%), polipos com hiperplasia em 17 casos (20,8%), sendo dez com hiperplasia simples sem atipias (12,2%) e sete com hiperplasia complexa sem atipias (8,6%). Em dois polipos foi encontrada neoplasia associada (2,4%). Para a analise estatistica foi empregado o teste do c2, corrigido por Yates. Correlacionou-se a histologia dos polipos com a presenca ou nao de sangramento genital (p=0,0056), o numero de polipos encontrados por mulher (p=0,921), e o tempo decorrido apos a menopausa (p=0,720). CONCLUSOES: polipos endometriais sao entidades frequentemente encontradas apos a menopausa, associando-se com baixa frequencia a hiperplasias e carcinomas endometriais e apenas o estudo histologico de todo especime nos permite afastar malignidade.