Polymorphism of the CYP2D6 gene in women with breast cancer treated with tamoxifen.

OBJECTIVE: To evaluate polymorphism frequency of the CYP2D6*4, *10, and * 17 alleles in women with breast cancer treated with tamoxifen. MATERIALS AND METHODS: Ninety-five women with estrogen and progesterone receptor-positive breast carcinoma were investigated from September to December 2013. A three-ml sample of peripheral blood was collected from each patient to analyze the presence of CYP2D6 *4, *10, and *17 allele polymorphism by specific polymerase chain reaction technique (PCR) for analysis of haplotypes *1, *4, *10, and *17, determined by studies of different single-nucleotide polymorphism (SNP). The data obtained were compiled and analyzed with the aid of Excel software 2010. RESULTS: The frequency of CYP2D6 alleles *4, *10, and *17 was 16%, 29%, and 2%, respectively, and haplotype * 1/*10 was shown in 22% of the women. The phenotype of intermediate metabolism occurred in 8% of women. CONCLUSIONS: The present study showed a deficiency in tamoxifen metabolism, characterized by intermediate metabolism in 8% of Brazilian women.

Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene.

OBJECTIVE: To evaluate Ki-67 antigen expression in the mammary epithelium of female rats in persistent estrus treated with raloxifene. MATERIALS AND METHODS: Forty-one Wistar-Hannover rats in persistent estrus induced by 1.25 mg of testosterone propionate were randomly divided into two groups: Group A (control, n = 21) in which the animals received only the vehicle (propylene glycol) and Group B (experimental, n = 20) in which the rats received 750 µg/day of raloxifene by gavage. After 21 days of treatment, all the animals were sacrificed and the first pair of abdominal-inguinal mammary glands was extirpated and fixed in 10% buffered formalin to investigate Ki-67 expression by immunohistochemistry. The data were analysed using Student's t-test (p < 0.05). RESULTS: The percentage of Ki-67-stained nuclei per 500 cells in the mammary epithelium was 42.33 ± 6.18 and 15.51 ± 3.71 [mean ± standard error of the mean (SEM)] in the control and experimental groups, respectively (p < 0.001). CONCLUSION: Raloxifene treatment significantly reduced Ki-67 expression in the mammary epithelium of rats in persistent estrus.

Assessment of quantitative and qualitative changes of proteoglycans and glycosaminoglycans in normal breast tissue during the follicular and luteal phases of the menstrual cycle.

The effect of sex hormones on extracellular matrix compounds, such as proteoglycans (PGs) and glycosaminoglycans (GAGs), in mammary tissue remains poorly understood. The elucidation of extracellular matrix component functions could clarify pathophysiological conditions, such as cyclical mastalgia (breast pain). The authors examined the quantitative and qualitative changes of PGs and GAGs in normal breast tissue during the follicular and luteal phases of the menstrual cycle. Twenty-eight eumenorrheic patients with benign breast nodules were divided into groups: Group A included 15 follicular patients and Group B included 13 luteal phase patients. Breast tissue adjacent to the nodules was biochemically analyzed to evaluate the types and concentrations of PGS and GAGs. The distribution of proteoglycans during the menstrual cycle was analyzed with immunofluorescence. PG concentrations were elevated (p < 0.01) during the luteal phase compared with the follicular phase, whereas the concentrations of GAGs did not differ significantly. Immunofluorescence revealed that decorin was mainly found in the intralobular stroma. PG concentrations were elevated during the luteal phase, likely due to the influence of sex hormones on macromolecular synthesis. The PG decorin was observed in normal breast tissue in the intralobular stroma. Although the concentration of GAGs, including dermatan and heparan sulfate, varied cyclically, the differences were not significant.

The relationship between estrogen receptor gene polymorphism and mammographic density in postmenopausal women.

OBJECTIVE: To assess the relationship between the presence of PVUII and XBAI polymorphisms in the estrogen receptor α gene and mammographic density in postmenopausal women. METHODS: For the present analysis, 189 postmenopausal women who had never used hormonal therapy and who did not have clinical or mammographic features were selected. Based on the ACR-BIRADS(®) 2003 classification, the mammographic density was determined by three independent readers (two subjective ratings and one computerized). Blood samples were available to extract DNA according to KIT GFX(®) protocol. PCR-RFLP was then used to identify the polymorphisms. RESULTS: There was a high degree of agreement among the three readers to determine the mammographic density (κ > 0.75). Sixty women (32%) had dense breasts and 129 (68%) had non-dense breasts. The PVUII polymorphism was found in 132 (69.8%) of 189 women, while the XBAI polymorphism was found in 135 (71.4%) women. Parity (p = 0.02) and body mass index (p < 0.0001) were associated with mammographic density. It was observed that, for the XBAI polymorphism, women with two mutated alleles were approximately 2.5 times more likely to be classified in the dense breasts group (p = 0.003) and the presence of both wild alleles was associated with fibroglandular tissue replacement by fat (p = 0.02). CONCLUSIONS: There was no significant association of the PVUII polymorphism in the estrogen receptor α gene with mammographic density (p = 0.34). However, the XBAI polymorphism was observed at a higher mutated homozygous frequency in women with dense breasts and there was an increased frequency of wild-type homozygous and heterozygous women with fat-replaced breasts (p = 0.01).

Association between estrogen receptor gene polymorphisms and breast density in postmenopausal women.

Objectives The aim of this study was to evaluate the association between clinical characteristics and polymorphisms HaeIII, MspI and XbaI of the estrogen receptor gene alpha with postmenopausal mammographic density. Methods A prospective study was performed with 120 women who were not users of hormones and had no identified breast lesions. All of them underwent bilateral mammography; the radiological density was determined by three independent observers, with two subjective evaluations based on the ACR-BIRADS(R) classification of mammographic patterns, 2003, and one computerized evaluation using the gray-scale histogram tool of the Adobe Photoshop(R) 7.0 software. Peripheral blood samples were obtained for DNA extraction, performed according to the GFX(R) Kit protocol (Amersham-Pharmacia). Polymerase chain reaction restriction fragment length polymorphism was carried out for an analysis of the polymorphisms present in intron 1 (HaeIII and XbaI) and in exon 1 (MspI) of the estrogen receptor gene. Results There was a high degree of concordance among the observers in the determination of mammary density (Kappa, Pearson and Spearman, p < 0.001). The associations of clinical characteristics with mammary density were: age (p = 0.04), body mass index (p < 0.0001) and age at menarche (p = 0.02). The relationship between the allele distribution of the polymorphisms and density was: XbaI (p = 0.02), HaeIII (p = 0.65) and MspI (p = 0.65). Conclusions Our data suggested that the polymorphism XbaI may be strongly related to mammographic density.

Paclitaxel in extensively pretreated nonseminomatous germ cell tumors.

Paclitaxel was given as a single agent to previously treated, cisplatin-refractory, and relapsed patients with metastatic nonseminomatous germ cell tumors (GCT). Fifteen patients received paclitaxel at 250 mg/m(2) as a 24-hour continuous intravenous infusion, repeated every 21 days. The regimen was premedicated to prevent hypersensitivity reactions. Patients were supported with granulocyte-colony stimulating factor until resolution of the neutropenia. Of 15 patients, I I patients had received at least three previous chemotherapy regimens; 80% of the patients were cisplatin-refractory at study entry. A total of 34 courses of paclitaxel were delivered. A serologic partial response and a partial remission for an overall response rate of 13.3% (95% C.I. 2-39%), with a median duration of 9.5 weeks, were achieved. Thirteen (86.6%) patients developed progressive disease. The toxicity of the regimen consisted of grade 3 4 neutropenia, with only one infectious complication and no life-threatening events. Nonhematologic toxicity was significant for progressive peripheral neuropathy in 8 patients. No hypersensitivity reactions or symptomatic cardiac toxicity occurred. In conclusion, paclitaxel in the dose and schedule given has minimal activity in this extensively treated, cisplatin-refractory group of patients with nonseminomatous GCT. Further investigation is warranted to find the role of paclitaxel in GCT by either selecting a better patient population and/or combining it with cisplatin, which would use the synergistic cytotoxicity that has been reported.

Experience with CA 15.3 as a tumor marker in breast cancer.

CA 15.3, a new tumor marker, is a glycoprotein antigen produced in greater amounts by breast tumor cells. It can be quantitatively detected, circulating in human serum or plasma, using an immunoradiometric assay with monoclonal antibodies. In order to evaluate the usefulness of the method and to determine the cut-off for metastatic disease, the CA 15.3 levels were determined in 78 patients (5 patients with breast fibroadenoma and 73 patients with breast cancer). The conclusions of the study are that the CA 15.3 is a useful parameter in the management of patients in different stages of the disease: levels above 36 U/ml are suggestive of metastasis, and above 86 U/ml are indicative of them. On the other hand, CA 15.3 does not seem to be helpful in the pre-operative differential diagnosis of breast lumps.

Hysteroscopic evaluation of the endometrium of post-menopausal patients with breast cancer before and after tamoxifen use.

OBJECTIVE: To evaluate by hysteroscopy and histopathology the influence of tamoxifen in the endometrium of post-menopausal women with previous breast cancer. METHOD: Out of 46 patients studied, 20 of them had been using tamoxifen for an average length of 12 months, and are still being followed-up. Hysteroscopy with endometrial biopsy was performed before and after the use of the drug. RESULTS: The prevalence of endometrial activity before and after this hormoniotherapy was the same, i.e. 10.0%, showing a non-significant variation. CONCLUSION: The hormoniotherapy with tamoxifen has not increased the endometrial proliferactive activity of postmenopausal patients with breast cancer. The most common hysteroscopical finding was numerous vesicles disseminated throughout the uterine cavity probably due to atrophy of the endometrium.

Effects of tamoxifen on the breast in the luteal phase of the menstrual cycle.

OBJECTIVES: The effect of tamoxifen on cyclic mastalgia and on chemoprophylaxis against breast cancer is little known, mainly due to the difficulties in studying the normal human gland. We proposed to evaluate the mitotic index and the nuclear volume of the lobule of women medicated with tamoxifen only during the luteal phase of the menstrual cycle in order to observe the effect of tamoxifen on the normal human mammary gland. METHODS: Twenty-four premenopausal women with fibroadenoma diagnosed via biopsy were studied. The phase of the cycle was determined by the date of menstruation and serum progesterone level in the luteal phase (> or = 3 ng/ml). The patients admitted to the study and were given written informed consent to participate in the investigation, which was previously approved of by the hospital Ethics Committee. Patients were divided at random into two groups: Group I consisted of 12 untreated women (control) and Group II consisted of 12 patients treated with 20 mg/day tamoxifen for 10 consecutive days beginning on the 13th day of the menstrual cycle. In both groups, the patients were submitted to biopsies of the nodule and of a 1-cm3 fragment of adjacent mammary parenchyma between the 23rd and 26th day of the cycle. The mitotic index (number of mitoses/1000 nuclei counted) and mean nuclear volume (mean of 10 nuclear volumes for each case) were measured. RESULTS: No mitosis was observed in Group II. There was a reduction in the mean nuclear volume in Group II (Mann-Whitney test). CONCLUSIONS: Tamoxifen, when administered only during the luteal phase of the menstrual cycle, significantly reduces the nuclear volume and mitotic activity of the epithelium. This data demonstrates an antagonistic action of tamoxifen on estrogen even when administered for short periods of time.

Morphological and ultrastructural aspects of the cyclical changes of human mammary gland during the menstrual cycle.

Analyzing the morphological and ultrastructural features of the mammary gland during the menstrual cycle, the authors observed that in the proliferative phase, the mammary gland consisted of few developed lobules and presented a relative absence of mitotic figures. In this phase, the great majority of the epithelial cells contained small, dark, irregular, rich heterochromatinic nuclei. The mammary stroma showed similar findings, presenting dense connective tissue with few enhanced fibroblasts. During the secretory phase, the lobules were well developed and mitotic figures were usually observed. In this phase, the great majority of the epithelial cells presented clear voluminous, rich euchromatinic nuclei; the stroma consisted of fibroblasts with voluminous nuclei full of euchromatin. There are evident modifications of the mammary tissue according to the phase of the menstrual cycle. The morphological and ultrastructural modifications indicate higher metabolical and mitotic activities during the secretory phase.

Malignant phyllodes tumor in the right breast and invasive lobular carcinoma within fibroadenoma in the other: case report.

CONTEXT: The malignant variety of the phyllodes tumor is rare. The occurrence of invasive lobular carcinoma within fibroadenoma is rare as well. DESIGN: Case report. CASE REPORT: A 58-year-old black female patient was referred to the Mastology unit of the Department of Gynecology, Federal University of São Paulo / Escola Paulista de Medicina, in February 1990, presenting an ulcerated tumor in the right breast with fast growth over the preceding six months. She was a virgin, with meno-pause at the age of 45 years and had not undergone hormone replacement treatment. The physical examination showed, in her right breast, an ulcerated tumor of 20 x 30 cm which was not adher-ent to the muscle level, multilobular and with fibroelastic consistency. The axillary lymph nodes were not palpable. The left breast showed a 2 x 3 cm painless, movable nodule, with well-defined edges, and fibroelastic consistency. We performed left-breast mammography, which showed several nodules with well-defined edges, the largest being 2 x 3 cm and exhibiting rough calcification and grouped microcalcifications within it. The patient underwent a frozen biopsy that showed a malignant variant of the phyllodes tumor in the right breast and fibroadenoma in the left one. After that, we performed a total mastectomy in the right breast and an excision biopsy in the left one. Paraffin study confirmed the frozen biopsy result from the right breast, yet we observed that in the interior of the fibroadenoma that was removed on the left, there was a focal area of invasive lobular carcinoma measuring 0.4 cm. The patient then underwent a modi-fied radical mastectomy with total axillary lymphadenectomy. None of the 21 dissected lymph nodes showed evidence of metastasis. In the follow-up, the patient evolved asymptomatically and with normal physical and laboratory examination results up to July 1997.

Leiomyoma of the breast. A case report.

A rare case of leiomyoma of the breast is reported, with a discussion of the clinical aspects and of the differential diagnosis. Excluding tumors originating from the areolar-papillary complex and the skin, this neoplasm is extremely rare, with only 11 cases reported so far. The histogenesis of the lesion is still controversial.

Morphometric analysis of fibroblasts of the mammary lobular stroma during the follicular and luteal phases of the menstrual cycle.

We determined the nuclear volume of fibroblasts of the normal mammary lobular stroma during the follicular and luteal phases of the menstrual cycle. Twenty patients aged 15 to 35 years and eumenorrheic for at least 6 months were randomly assigned to 2 groups, i.e., 10 women in the follicular phase and 10 in the luteal phase. The nuclear volume was 34.4 micron 3 and 98.8 micron 3 for the follicular and luteal phases, respectively, with the difference being statistically significant (p < 0.05). These data suggest a higher metabolic activity in the mammary intralobular stroma during the luteal phase of the menstrual cycle, probably due to a synergistic action of estradiol and progesterone.

Correlación entre el grado de neoplasia intraepitelial cervical y la expresión dual de p16 Y ki67, en biopsia de cuello uterino en el Hospital Santa Rosa de Lima-Perú año 2016 / Correlation between the degree of cervical intraepitelial neoplasis (nic) and the dual expression of p16 and ki67, in uterine neck biopsy in Santa Rosa Hospital of Lima-Peru year 2016

Objetivo: Determinar la frecuencia de coexpresión dual del p16 (INK4a) y ki67 en los diferentes grados de las neoplasias intraepiteliales cervicales (NIC) y su contribución molecular para diagnóstico y pronóstico. Material y Metodos: El diseño es descriptivo, prospectivo transversal. Se revisaron 20 biopsias de cérvix embebidas en parafina, que se encontraban en el archivo de Anatomía Patológica de un Hospital tercer nivel de Perú entre Enero y Diciembre del 2016, cuyas láminas portaobjeto fueron coloreadas con hematoxilina-eosina y reportadas como neoplasia intraepitelial (NIC) I, II, III. Se realizaron nuevos cortes histológicos de 3 micras para estudios complementarios usando la técnica de inmunohistoquímica dual. Las células con positividad para p16 mostraron tinción citoplasmática y/o nuclear de color rojizo y aquellas con positividad para ki67 mostraron coloración nuclear marrón. Resultados: Los 20 casos de neoplasia intraepitelial se dividen en: 06 casos de NIC I; 11 casos de NIC II y 03 casos de NIC III. La tinción dual para el Ki67 y p16 se realizó a los 20 casos, esto mostró una coexpresión en el 85% y ausencia de expresión dual en 15%. Se observó tinción en los diversos grados de NIC predominando el grado 1+ en un 50%. En el grado 2+ y 3+ la expresión fuerte en NIC II y NIC III respectivamente. La expresión del Ki67 se dio en todos los diferentes grados de NIC. Conclusiones: La coexpresión del Ki67 y p16 es de 85% en los diversos grados de NIC, siendo frecuente en el grupo de NIC II. (AU)

Objetive: To determine the frequency of dual coexpression of p16 (INK4a) and ki67 in the different degrees of cervical intraepithelial neoplasia (CIN) and its molecular contribution for diagnosis and prognosis. Material And Methods: the design is descriptive, prospective and cross-sectional. 20 cervix biopsies embedded in paraffin have been reviewed. These slides gathered from January 2016 to December 2016 were located in the records of Anatomical Pathology Department of a third level Hospital in Peru. The slides were stained with hematoxylin-eosin and reported as intraepithelial neoplasia (CIN) I, II, III. New histological sections of 3 microns were made for complementary studies by using dual immunohistochemistry technique. The cells with positivity for p16 showed cytoplasmic and / or nuclear reddish staining and those with positivity for ki67 showed brown nuclear coloration. Results: The 20 cases of intraepithelial neoplasia are divided into: 06 cases of CIN I; 11 cases of CIN II and 03 cases of CIN III. Dual staining for Ki67 and p16 was performed in all 20 cases. This showed coexpression in 85% and absence of dual expression in 15%. Staining was observed in the different grades of CIN, with predominance in grade 1+ ( 50%). In grade 2+ and 3+ the expression was strong in CIN II and CIN III respectively. The expression of Ki67 occurred in all the different grades of NIC. Conclusions: The coexpression of Ki67 and p16 is 85% in the different grades of CIN, being frequent in the group of CIN II. (AU)

Analysis of human mammary fibroadenoma by Ki-67 index in the follicular and luteal phases of menstrual cycle.

OBJECTIVES: Fibroadenoma is the most common benign mammary condition among women aged 35 or younger. Expression of Ki-67 antigen has been used to compare proliferative activity of mammary fibroadenoma epithelium in the follicular and luteal phases of the menstrual cycle. MATERIALS AND METHODS: Ninety eumenorrheic women were selected for tumour excision; they were assigned to either of the two groups, according to their phase of menstrual cycle. At the end of the study, 75 patients with 87 masses were evaluated by epithelial cell Ki-67 expression, blind (no information given concerning group to which any lesion belonged). RESULTS: Both groups were found to be homogeneous relative to age, menarche, body mass index, previous gestation, parity, breastfeeding, number of fibroadenomas, family history of breast cancer and tabagism. Median tumour size was 2.0 cm and no relationship between proliferative activity and nodule diameter was observed. No typical pattern was observed in the expression of Ki-67 in distinct nodules of the same patient. Average values for expression of Ki-67 (per 1000 epithelial cells) in follicular and luteal phases were 27.88 and 37.88, respectively (P = 0.116). CONCLUSION: Our findings revealed that proliferative activities in the mammary fibroadenoma epithelium did not present a statistically significant difference in the follicular and luteal phases. The present study contributes to clarifying that fibroadenoma is a neoplasm and does not undergo any change in the proliferative activity during the menstrual cycle.

Proximal humeral ossification center of the fetus: time of appearance and the sensitivity and specificity of this finding.

Ultrasonographic examinations were performed on 1,000 pregnant women between the 28th and 40th weeks of gestation who had been seen at a private clinic. For all women the date of last menstruation was known, and none had obstetrical or clinical-surgical complications. All were singleton pregnancies, and cases of fetal malformation were excluded from the study. The proximal humeral epiphyseal ossification center was visualized significantly frequently after the 38th week (P < 0.001). With respect to a gestational age of more than 38 weeks, its specificity was elevated (99%), as also were its positive (91%) and negative (93%) predictive values. Its sensitivity, however, was low (58%). Routine determination of the proximal humeral ossification center is a valuable aid for the estimation of gestational age in the third trimester.

[Comparison between pelvic endovaginal and transabdominal sonography in the measurement of the uterus and ovaries]. / Comparação entre a ecografia pélvica endovaginal e a transabdominal na mensuração do útero e dos ovários.

PURPOSE: this study compares endovaginal sonography to the transabdominal ultrasound in terms of their accuracy in the measurement of the uterus and ovaries. TYPE: prospective study. PLACE: São Paulo, SP, Brazil. PATIENTS: Forty-nine women were included in the study in order to assess the clinical suspicion of pelvic mass or because they had vaginal bleeding in initial gestation. The patients were submitted to transabdominal sonography and later to endovaginal ultrasound (5.0mHz transducer). Uterus and ovaries were measured. RESULTS: the volume of 27 left and 29 right ovaries was measured. Echographic hysterometry was performed in 49 women and statistically compared. The Wilcoxon rank sum test failed to show any significant difference between the two sonographic techniques in the measurement of uterus and ovary volume. CONCLUSIONS: endovaginal ultrasound presents several advantages over transabdominal scanning. Patient's discomfort is minimum because vesical repletion is not called for. Imaging resolution is magnified and morphologic details are clearly seen because the transducer is in close contact with pelvic viscera. Results do not evidence any significant difference in the uterine measurements with either method, and the statistical difference in ovarian volume was not significant either.

[Apocrine gland cell carcinoma of the vulva. Report of a case]. / Carcinoma de células apócrinas da vulva. Relato de um caso.

The authors have described a rare case of apocrine glands carcinoma of the vulva in a 58-year old black patient. She underwent radical vulvectomy and radiotherapy. After 8 months, she did not show any local or systematic recurrences.

Histological study of the endometrium in menopausal women with breast carcinoma.

The first reference concerning the multiple primary malignant neoplasms was made by Bilroth, 1880 and since then a large number of studies have been published. Furthermore, an increasing incidence of simultaneous cancers are currently observed (16,18). At the same time, several retrospective populational studies evidenced the association between breast and endometrium carcinoma. It is well known that both uterus and breast are hormone-dependent organs and are likely to be influenced by the same oncogenic stimulus, either of endocrine nature or dietary origin (2, 3, 14). The risk of developing endometrial carcinoma is higher in patients already affected by breast neoplasm and is much more evident in older women within the first five years following the diagnosis of breast tumor. Conversely, patients with endometrial carcinoma may present a second mammary neoplasm and the relative risk is around 2.0 (1, 17). Although the existence of a correlation between these two primary malignant neoplasms is clearly observed, the absence of systematic studies directed to the screening of endometrial cancer in women with breast carcinoma is surprising. This study deals with histological analysis of the endometrium of postmenopausal patients with breast cancer and aims to determine the possible changes that might have occurred in the onset of the disease.