HPV genotyping and p16 expression in Xingu Indigenous Park, Brazil.

The association between high-risk human papillomavirus (HPV) genotypes and p16 expression in indigenous women from the Xingu Indigenous Park, Brazil, was unknown. This study evaluated p16 expression in women with a histological diagnosis of cervical intraepithelial neoplasia (CIN) 3 or higher and correlated this expression with HPV genotypes to determine possible discrepancies in the expression of this marker. We evaluated 37 previously collected samples with different HPV genotypes and high-grade lesions diagnosed based on cytology, histology, and colposcopy. Immunohistochemical analysis was performed using paraffin-embedded tissue sections and the CINtec® Histology Kit. p16 protein expression was investigated by immunostaining with an anti-p16 antibody. HPV genotyping was performed by reverse hybridization. The age of the study population ranged from 22-75 years (43.81 ± 15.89 years) and parity ranged from 1-11 (5.92 ± 2.58). Thirteen different HPV genotypes were found using the INNO-LiPA kit. Single and multiple infections by HPV were found with prevalence of single infections (P = 0.029). Comparison between HPV genotype and simple or multiple infections was highly significant; it was observed more HPV 52 followed by HPV 16 in single infections (P < 0.001). p16 expression was predominantly diffuse, which was observed in 91.7% of lesions, whereas 8.3% were focal (P < 0.001). HPV 52, HPV 16 and 31 were the most prevalent HPV types in high-grade CIN in these indigenous women. Diffuse p16 expression in high-grade CIN was not influenced by the viral genotype; however, more studies are necessary to further our understanding of this restricted group.

Immunoexpression of HPV 16/18 E6 and E7 oncoproteins in high-grade cervical squamous intraepithelial lesions in HIV-positive women.

The aim of this study was to assess the immunoexpression of human papillomavirus genotypes 16 and 18 (E6 and E7) oncoproteins in cervical high-grade squamous intraepithelial lesions (HSIL) of human immunodeficiency virus (HIV)-positive women. These results were also compared to the persistence and/or recurrence of lesions after loop electrosurgical excision procedure. Cervical samples from 158 patients were divided into three groups according to the presence or absence of HSIL in women who were or were not HIV-positive. By using the tissue microarray technique, immunohistochemistry was performed to analyze the expression of HPV 16/18 E6 and E7 oncoproteins. Cervical samples from 95 HIV-positive women and 63 HIV-negative women were studied. A statistically significant difference was found in the immunoexpression of E6 and E7 oncoproteins in samples from HIV-positive women with HSIL and that of women with non-neoplastic tissue (P < 0.001). There was also a statistically significant correlation between the immunoexpression of E6 (P = 0.012) and E7 (P < 0.001) oncoproteins in lesion persistence among HIV-positive women. Within the limitations of this study, the immunoexpression of HPV 16/18 E6 and E7 oncoproteins may have prognostic value regarding lesion persistence in HIV-positive women.

Depth of glandular crypts and its involvement in squamous intraepithelial cervical neoplasia submitted to large loop excision of transformation zone (LLETZ).

BACKGROUND: The authors aimed to confirm the depth of six mm in order to achieve an optimal eradication of the lesion. MATERIALS AND METHODS: This is a retrospective observational study of 94 cervical surgical pieces from women aged 17 to 22 years with a cyto-colpo-histopathological diagnosis of high-grade squamous cervical intraepithelial neoplasia (CIN II and/or CIN III) submitted to large loop excision of transformation zone (LLETZ). The glandular crypts and margins, both exposed or not to CIN, were assessed. The compromise and the maximum depth of the glandular crypts were noticed. RESULTS: After LLETZ, 23 (24.47%) cases presented a neoplasic impairment of endocervical margin and ten (10.64%) of the ectocervical margin. The largest noticed crypt measured 4.500 mm and the shortest 0.100 mm, with an average of 2.148 mm. CONCLUSIONS: Squamous CIN more frequently show the exposure of surgical margins to LLETZ. The deeper location of glandular crypts in the cases studied was 4.500 mm, while the largest neoplastic extension was 3.000 mm.The therapeutic method depends on this knowledge.

Immunohistochemical expression of granzyme B and vascular endothelial growth factor (VEGF) in normal uterine cervices and low and high grade squamous intraepithelial lesions.

PURPOSE: This study aimed to evaluate the immunoexpression of granzyme B and vascular endothelial growth factor (VEGF) in the variants of cervical squamous intraepithelial neoplasia. METHODS: Granzyme B immunohistochemical expression was studied in the epithelium, stroma and in both the epithelium + stroma of 142 fragments of uterine cervix; there were 34 grade 1 cervical intraepithelial neoplasias (CIN 1), 36 grade 2 cervical intraepithelial neoplasias (CIN 2), 33 grade 3 cervical intraepithelial neoplasias (CIN 3) and 39 uterine cervix fragments without abnormalities - control group. Immunoexpression of VEGF was studied in 160 uterine cervix fragments, with 43 grade 1 cervical intraepithelial neoplasias (CIN 1), 33 grade 2 cervical intraepithelial neoplasias (CIN 2), 31 grade 3 cervical intraepithelial neoplasias (CIN 3) and 53 uterine cervix fragments without abnormalities--control group. RESULTS: In the stroma, immunoexpression of granzyme B in grade 1 cervical intraepithelial neoplasias was smaller than in grade 3 cervical intraepithelial neoplasias. High VEGF immunoexpression was found in grade 3 cervical intraepithelial neoplasias while it was low in grade 1 cervical intraepithelial neoplasias and in the control group. CONCLUSION: The higher the severity of the cervical intraepithelial lesion, the higher the immunoexpression of granzyme B. A progressive increase in VEGF immunoexpression was found in the intense grade, according to the severity of the cervical intraepithelial neoplasia.

Prognostic markers of low-grade squamous intraepithelial lesions: the role of topoisomerase IIalpha and active caspase-3.

PURPOSE: To study the relationship between topoisomerase IIalpha, active caspase-3 expressions and HPV DNA in uterine cervices with low-grade squamous intraepithelial lesions (LSIL). METHODS: Forty women with LSIL and 32 without cervical neoplasia diagnosed through cytologic and histopathologic examination were evaluated regarding topoisomerase IIalpha and active caspase-3 expressions and HPV DNA detection using PCR (GP5/GP6) in cervicovaginal smears. RESULTS: The mean percentage of cells immunomarked by topoisomerase in the group with LSIL was 11.62% while in the control it was 4.13% (p < 0.0001). In the presence of HPV DNA, topoisomerase expression was higher in the group with productive viral infection than in nonneoplastic tissue (p = 0.004). Caspase-3 expression was observed in 17 patients with LSIL (42.5%) and in five without cervical neoplasia (15.63%). CONCLUSION: The use of topoisomerase IIalpha and active caspase-3 in cervical biopsies may help to define the prognosis of HPV cervical infection.

Use of dopamine agonist pergolide in outpatient treatment of cocaine dependence.

The dopamine agonist pergolide was evaluated in the treatment of 42 men who manifested cocaine dependence in a single-blind, 4-week-long placebo-controlled study, during 1998-1999 in São Paulo, Brazil. The patients were randomly assigned to two groups: the first group received pergolide (0.05-0.2 mg per day) and the second group received placebo (one to four tablets per day). Urine toxicology screens were obtained. The groups were compared in terms of depressive symptoms, "craving," use of cocaine, side effects of medications, results of urine tests, and retention in treatment. At 3 months' follow-up, the participants were reassessed. No differences were found between the two groups.

Granzyme B as a prognostic marker of cervical intraepithelial neoplasia.

Granzyme B (GrB) is a serine protease synthesized in T lympocytes (CTL), released after T-cell activation resulting from exogenous stimulation. With perforin, GrB discharges apoptotic signals to a target cell and therefore constitutes a marker to identify activated CTL. We aimed to quantify GrB expression by immunohistochemistry staining in 12 tissue fragments of cervical carcinoma, 33 cervical intraepithelial neoplasias treated by LLTEZ and nine cervical pieces without disease. Activated cytotoxic lymphocyte mean values (20 HPF-400x) in both epithelial and stromal pars were 7.11 cells in tissue without neoplasia, 33.45 cells in cervical intraepithelial neoplasia and 139.75 cells in carcinoma samples, with a statistical difference between them. Comparative analysis in the CIN group showed an expressive difference between cases with disease recurrence (19.28 cells) and without recurrence (37.26 cells). Thus, the relation between number of activated CTLs found at the moment of treatment and clinical evolution determined in this study, suggest GrB use as a prognostic marker.

Angiogenesis in squamous intraepithelial neoplasia of the uterine cervix in HIV-seropositive women.

OBJECTIVES: This study aimed to quantify angiogenesis in squamous intraepithelial lesions of the uterine cervix in seropositive HIV patients as well as to establish a relationship between vascular density and variations in the CD4+ lymphocyte titer and the viral load of human immunodeficiency virus (HIV). METHODS: 125 patients, 55 HIV seropositive and 70 seronegative, were allocated with respect to grade of squamous intraepithelial lesion (SIL). The obtained samples were stained with an immunohistochemical marker for CD34 antigen and vessel counts were performed in ten consecutive fields at 400x magnification. The seropositive HIV patients were distributed into groups according to the CD4+ index and HIV viral load. RESULTS: Seropositive HIV patients presented a higher mean vascular density (MVD) than the control group, even in the absence of cervical intraepithelial lesions. High- and low-grade lesions in the presence of HIV seropositivity presented higher MVD than that found in seronegative HIV patients. There was no significant variation in the MVD and CD4+ count ratio or viral RNA-HIV load, except for high-grade (H)SIL. CONCLUSIONS: Infection with HIV influenced angiogenesis of uterine cervix in the presence of squamous intraepithelial lesions and more significantly in HSIL.

Histologic studies of the effects of circulating hot saline on the uterus before hysterectomy.

STUDY OBJECTIVE: To evaluate a new device for endometrial ablation. DESIGN: (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Thirty-two women scheduled for hysterectomy. INTERVENTIONS: Endometrial ablation and hysterectomy. MEASUREMENTS AND MAIN RESULTS: The new device for endometrial ablation was evaluated by studying depth of necrosis after staining for the oxidative enzyme NADH. Uniform endomyometrial necrosis was achieved at a depth of 2 to 4 mm with 90 degrees C saline circulated for 10 minutes. CONCLUSION: The procedure was successful in all patients, and there were no adverse clinical sequelae. (J Am Assoc Gynecol Laparosc 6(3):269-273, 1999)

Evaluation of Hydro ThermAblator for endometrial destruction in patients with menorrhagia.

OBJECTIVE: To describe endometrial destruction with the Hydro ThermAblator. DESIGN: (Canadian Task Force classification II-1). SETTING: University-affiliated hospital. PATIENTS: Twenty-six women with uncontrolled menorrhagia. INTERVENTION: Endometrial ablation with saline heated to 90 degrees C and circulated in the uterine cavity for 10 minutes under hysteroscopic control. MEASUREMENTS AND MAIN RESULTS: All patients tolerated the procedure well, all but one was satisfied with the results, and there were no significant side effects or complications. Seventy-seven percent of women were either amenorrheic or hypomenorrheic after 6 months, and 88% were amenorrheic or hypomenorrheic after 1 year, including one patient who was treated a second time; 87.5% were amenorrheic or hypomenorrheic at 18 months. One woman had persistent menorrhagia and was treated by hysterectomy. CONCLUSION: In this study, endometrial ablation was performed successfully with the Hydro ThermAblator in 25 of 26 women. (J Am Assoc Gynecol Laparosc 6(3):275-278, 1999)

Morphologic and morphometric aspects of the endometrium of postmenopausal women before and after cyclic oestrogen replacement treatment.

OBJECTIVE: The aim of the study is to observe the morphology and morphometry of the endometrium of postmenopausal women treated with cyclic conjugated oestrogens. STUDY DESIGN: Three groups of nine postmenopausal women received cyclic conjugated oestrogens for 21 days (with a seven-day pause) during six months. The endometrial specimens were obtained using a modified Novak suction curet, in the second or third day of the period of drug washout. The slides were stained with haematoxylin and eosin (H.E.) in order to measure epithelial height and determine the gland/stroma ratio. RESULTS: Morphologic examination showed that single daily doses of 0.3 mg of conjugated oestrogens caused discrete endometrial proliferation after three and six months of treatment. However, a more intense effect was observed in women receiving doses of 0.625 and 1.25 mg/day of the hormone, in the same period. Morphometric study revealed significant increases both in epithelial thickness and in the gland-stroma ratio, specially in women receiving higher doses of the conjugated oestrogen (0.625 and 1.25 mg/day). CONCLUSIONS: We concluded that there were marked proliferative alterations without atypias in the endometrium of women that received 0.625 and 1.25 mg of conjugated oestrogens during six months.

Síndromes delirantes de falso reconhecimento: curiosidade ou chave para um novo conhecimento psiquiátrico? / Misidentification syndromes: curiosity or a key for a new psychiatric knowledge?

Objetivos: Partindo das definiçoes das Síndromes Delirantes de Falso Reconhecimento, tendo como principais modelos as Síndromes de Capgras e Frégoli, os autores fizeram uma revisao dos aspectos psicopatológicos envolvidos nesses quadros. Métodos: Foi realizado um levantamento da Literatura fenomenológica clássica e posterior adaptaçao de um modelo neuropsicológico que aprofunde o entendimento desses quadros, geralmente negligenciados pela literatura ou tratados como extravagências de psicopatólogos antigos. Conclusoes: O entendimento das Síndromes Delirantes de Falso Reconhecimento, no entender dos autores, pode ser de grande valia para formar novos conceitos no estudo dos quadros psiquiátricos delirante-alucinatórios maiores, principalmente das correlaçoes entre Sensaçao, Percepçao e Atribuiçao de Significado.

Invasive ductal carcinoma, fibroadenoma and adjacent breast stroma angiogenesis: an immunohistochemical and morphometrical study.

The authors investigate the differences between the angiogenesis that occurs in malignant and benign breast tumours (10 specimens of invasive ductal carcinoma and 10 specimens of fibroadenoma) as well as in the adjacent breast stroma of these women highlighting the microvessels by staining their endothelial cells for factor VIII immunohistochemically. The number of vessels counted in invasive ductal carcinoma was significantly higher than the number of vessels counted in fibroadenoma and in the adjacent breast stroma. There was no difference between the number of vessels counted in the fibroadenoma and its adjacent stroma. Even though the patients with invasive ductal carcinoma showed a adjacent stroma with a higher number of vessels, the difference was not significant when compared with the adjacent stroma obtained from patients with fibroadenoma.

Ultrastructural aspects of the remodeling process of the Corpus albicans in the recent postmenopausal period.

To study the cytophysiology of the corpus albicans in the recent postmenopausal period, the authors analyzed the ovarian ultrastructure of ten patients submitted to oophorectomy due to non-malignant gynecological diseases. Evidence of a remodeling process with connective tissue substitution of the corpora albicantia was observed. The remodeling process appears to depend on the activity of three essential cell types; the fibroblasts, which provide collagen synthesis; the macrophages, which phagocytize the flaky material; and the myofibroblasts, mainly located in the peripheral region of the corpora albicantia, which may have a retracting action on the remodeling site of the corpus albicans.

Morphological and biochemical action of dipyrone on rat placenta.

1. The morphological and biochemical action of dipyrone (N-[2,3-dimethyl-5-oxo-1-phenyl-3-pyrazolin-4-yl]-methylamino methanesulfonate, sodium monohydrate) on the placenta of albino rats was studied by means of karyometry of trophoblastic giant cells and by determinations of DNA, RNA and total protein contents. 2. The animals were treated with a single daily dose of 50 mg/kg body weight during 5 different periods: from the 9th to the 12th, 11th to the 14th, 13th to the 16th, 15th to the 18th or 17th to the 20th day of pregnancy. 3. Karyometric results showed that the nuclear volumes of placental cells in rats treated with dipyrone during the first 3 periods were significantly greater than in control animals and that, closer to term, no differences were observed in this regard. Only the animals treated from the 9th to the 12th day of pregnancy had higher placental contents of DNA, RNA and protein than the corresponding controls. 4. Our results showed that dipyrone had a blocking effect on placental cell division which occurs mainly in the initial steps of placental development.

Morphological and morphometrical features of the corpus albicans in the course of the postmenopausal period.

Studying the morphological and morphometrical aspects of corpora albicantia at different phases of the postmenopausal period, the authors observed that, until five years after menopause, the fibroblasts were developed, and later in this period of life, the number and activity of these cells are diminished. It is suggested that the description of corpora albicantia in atrophic ovaries of postmenopausal women occurs owing to a decrease of remodelling fibroblastic and macrophagic activity, or connective tissue substitution, made by these cells under physiological conditions.

Action of diclofenac sodium on the reparation process of the surgically-perforated myometrium of pregnant albino rats: morphological and morphometrical evaluation.

1. The action of a non-hormonal anti-inflammatory agent on the reparation process of the surgically-perforated myometrium of pregnant albino rats was histologically evaluated. 2. On the 14th day after perforation, the muscular layers were still interrupted. 3. The morphological study was inconclusive. A morphometrical analysis, however, demonstrated that there was a meaningful diminution in the number of leucocytes and collagen fibers and an increase in the number of fibroblasts, probably with low synthetical power, in the group treated with diclofenac sodium. All these data indicate a delay in the reparation process of the uterine musculature.

Premature ovarian failure: morphological and ultrastructural aspects.

The authors documented by means of light and transmission electron microscopy that the ovaries of women with premature ovarian failure (POF) displayed dense connective tissue and rare corpora albicantia. Eight of the ten studied cases did not present ovarian follicles; in two cases, it was verified the presence of ovarian follicles, atypical primordial follicles and in one case, a corpus luteum was identified (after stimulation with exogenous gonadotrophin). Regarding the ultrastructural analysis, it was noted that the fibroblasts were united one to each other by cellular prolongations that formed a woof, constituting a cellular syncicius.

[Human colostrum and milk cells]. / Células do colostro e leite humanos.

This review dwells on the major cell defense factors of human colostrum and milk: phagocytes, lymphocytes, epithelial cells, and natural-killer cells (NK). Other elements are also presented, of humoral origin, which take part in such fluid. The cells are morphofunctionally characterized and reasons are given that make it more difficult for a detailed study (great variability, atypical morphology, presence of fatty globules). Finally, the function of maternal milk is stressed as a transmitter of systemic immunity and of nutritional elements to the neonate, therefore the importance of breastfeeding.