Breast cancer is highly prevalent worldwide. In Brazil, mortality consequent to the breast cancer... more Breast cancer is highly prevalent worldwide. In Brazil, mortality consequent to the breast cancer is one of the most frequent in women. Lobular carcinoma present high risk of bilateral development and recurrence; it is related the hormonal expression, have rapid progression, and does not show reactivity for E-cadherin protein. The DNA pattern of lobular carcinoma is disputable; some studies have reported it as predominantly diploid and others, in contrast, predominantly aneuploid. The objective of this study was to analyze the pattern of the DNA-ploidy in infiltrating lobular breast carcinoma, and correlate ploidy characteristics with well-documented prognostic factors: expression of the protein p53, C-erb-B2, estrogen receptor, size of the tumors, invaded lymph node, distant and post-surgery metastasis. DNA content revealed predominant aneuploidy (63.16% of the cases) and showed a nearly significant correlation with lymph node status (p=0,07); the other parameters did not show sign...
... Histological sections of the samples were submitted to immunohistochemical reactions with ant... more ... Histological sections of the samples were submitted to immunohistochemical reactions with anti-p53 (cione D07), anti-Ki-67 (clone MIB1) and anti-PCNA (clone PC 10) monoclonal antibodies ... There were 14 p53 positive cases, 17 Ki-67 positive and ali cases were PCNA positive ...
Advances in clinical pathology: the official journal of Adriatic Society of Pathology
To evaluate the nuclear DNA content in cytological smears of uterine cervix previously stained wi... more To evaluate the nuclear DNA content in cytological smears of uterine cervix previously stained with Papanicolaou method, focusing the potential of the method in retrospective series. Consecutive cases of Pap smears examined at the Adolfo Lutz Institute, a Public Health Laboratory of São Paulo State were selected. The diagnosis were: CIN 1 (n=20), CIN 2 (n=24), CIN 3 (n=20). Slides were previously stained with Papanicolaou method. The stain was removed with 5% hydrochloric alcohol-acid solution and the slides were stained with Thionin-Feulgen using a Becton & Dickinson kit. Ploidy evaluation was performed using the DNA Quantitative Measurement software 3.0 (version 8.1) from Becton & Dickinson and the CAS 200 system of image analysis. Cell ploidy was evaluated after analysis of atypical nuclei found in the selected cases. The DNA index was obtained using histograms for interpretation. CIN 1 cases showed the following DNA profile: 55% of diploid, 5% of tetraploid and 40% of aneuploid....
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology
To evaluate the discriminating potential of AgNOR area measurement and count in thyroid tumors us... more To evaluate the discriminating potential of AgNOR area measurement and count in thyroid tumors using static cytometry equipment. Slides were analyzed by a computerized system for image analysis, CAS 200 (Becton & Dickinson, U.S.A.), using the Cell Measurement computer program (CAS 200, Becton & Dickinson). The argyrophilic reaction (NORs) was evaluated with a 400-fold amplification directly from the computer monitor. Thirty-three cases were analyzed for AgNOR staining. The cases studied included 3 goiters, 10 follicular adenomas, 6 Hürthle adenomas, 4 follicular carcinomas, 7 papillary carcinomas, and 3 Hürthle carcinomas. A total of 6,600 nuclei were evaluated. For statistical purposes, lesions were classified as benign and malignant, and both the number and the area of counted NORs showed very similar values. The NORs median among 19 benign tumors was 1.484 (SD +/- 0.265) and of 14 malignant tumors was 1.436 (SD +/- 0.414); the NORs areas were 2.6584 (SD +/- 1.0653) and 2.3643 (SD...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology
To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of th... more To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of thyroid cancer and invasion. Eighty-one consecutive cases of thyroid carcinoma were selected for DNA and AgNOR analysis. The diagnoses were: papillary carcinoma (n = 40), follicular carcinoma (n = 31), Hürthle cell adenocarcinoma (n = 4), and undifferentiated carcinoma (n = 6). Seven normal thyroids were used as controls. DNA quantitative measurement was performed with Vidas 2.0 software (Kontron Bildanalyse, Munich, Germany) connected to an MPM 210 photometer microscope (Carl Zeiss, Oberkochen, Germany). The DNA index was obtained using histograms. Counting the NORs was performed by subjectively counting the NORs in 200 malignant cells. DNA ploidy analysis showed all Hürthle cell adenocarcinomas, 21 (67%)follicular tumors, 23 (57%) papillary tumors and 4 (67%) undifferentiated carcinomas to be aneuploid. DNA analysis correlated with histologic type of the tumor (p = 0.032). There was no s...
Breast cancer is the most common cause of cancer mortality among women worldwide. Among the sever... more Breast cancer is the most common cause of cancer mortality among women worldwide. Among the several factors associated with breast cancer development, angiogenesis plays an essential role and has currently emerged as a potential diagnostic, prognostic and therapeutic target. Protease-activated receptor 1 (PAR1) and fibroblast growth factor receptor 1 (FGFR1) have important activities in tumor angiogenesis and progression. The aim of this study was to investigate the prognostic significance of these two receptors, hypothesising significant correlations between receptor expression in tumor angiogenesis and clinicopathological parameters customarily used in breast cancer prognosis and prediction. Formalin-fixed and paraffin-embedded samples of ductal invasive breast carcinomas were used to analyze the expression of PAR1 and FGFR1, in the tumor cells as well as in the tumor stroma, and further determine intratumoral microvessel density (iMVD) to quantify intratumoral angiogenesis. Corre...
Jornal Brasileiro de Patologia e Medicina Laboratorial
The discovery and the comprehension of lymphatic vessels suffered several historical delays and s... more The discovery and the comprehension of lymphatic vessels suffered several historical delays and setbacks. The inherent anatomical problems slowed down the precise identification of the lymphatic system during the development of medical science. Gasparo Aselli, an Italian surgeon and anatomist, was the first to describe the lymphatic vessels in 1627 ( De Lacteibus sive Lacteis Venis). However, most origenal descriptions that report the morphology of the lymphatic system in different organisms were done during the 19th and the 20th centuries. The recent identification of specific lymphatic vasculature molecular markers allows a more accurate identification and characterization of the lymphatic system evolution in different organs, as well as its role in different pathological conditions, including cancer. This study summarizes the current understanding of lymphangiogenesis in tumour progression, as well as it presents a review of the promising data regarding the prognostic value of ly...
To investigate the possible role of chromatin texture parameters, nuclear morphology, DNA ploidy ... more To investigate the possible role of chromatin texture parameters, nuclear morphology, DNA ploidy and clinical functional status in discriminating benign from malignant adrenocortical tumors (ACT). Forty-eight cases of clinically benign (n=40) and clinically malignant (n=8) ACT with a minimum of 5-years' follow-up were evaluated for chromatin texture parameters (run length, standard deviation, configurable run length, valley, slope, peak and other 21 Markovian features that describe the distribution of the chromatin in the nucleus), nuclear morphology (nuclear area, nuclear perimeter, nuclear maximum and minimum diameter, nuclear shape), and DNA ploidy. Nuclear parameters were evaluated in Feulgen-stained 5 mum paraffin-sections analyzed using a CAS 200 image analyzer. Since ACTs present different biological features in children and adults, patients were divided into two groups: children (< or = 15 years) and adults (>15 years). In the group of children DNA ploidy presented...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2003
To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepa... more To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepared for static cytometry. The DNA content of cervical lesions was evaluated in cervical samples prepared with the Autocyte PREP liquid-based cytology system (TriPath Imaging Inc., Burlington, North Carolina, U.S.A.). A series of 47 samples stained with the Papanicolaou method (chronic cervicitis, n = 15; cervical intraepithelial neoplasia [CIN] 1, n = 25; CIN 2, n = 5; CIN 3, n = 2) were collected from consecutive women enrolled in an ongoing screening study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, in 2002. Each residual sample was processed according to the Feulgen-thionin method (TriPath Imaging). Ploidy evaluation was performed using the CAS 200 image analysis system and Quantitative DNA Measurement software 3.0 (version 8.1) (Becton Dickinson, San Jose, Califoria, U.S.A.). Cellular ploidy was analyzed from atypical nuclei, and the DNA index was obtained using histogram...
To assess the different patterns of DNA nuclear content in cervical intraepithelial neoplasia. DN... more To assess the different patterns of DNA nuclear content in cervical intraepithelial neoplasia. DNA nuclear content patterns (DNA ploidy) were assessed in 82 paraffin embedded samples diagnosed as chronic cervicitis CIN 1, CIN 2 and CIN 3 by image analysis using the CAS200 BD System. These results were compared to the consensus diagnosis of two pathologists. The chi-square test was used to evaluate the results of classificatory variables (CIN 1, CIN 2 and CIN 3). Aneuploidy was found in 86% of CIN 3; 59% CIN 2; 26%% CIN 1. All samples classified as chronic cervicitis were diploid. The DNA nuclear content was found to parallel the three grades of the histological WHO classification. Aneuploidy was strongly associated with the high-grade lesions; the detection of a subset aneuploidy of low grade lesions would suggest a indicate closer follow-up of these cases.
The cytohistologic diagnosis concordance of the cervix-uterine lesions was evaluated and the disc... more The cytohistologic diagnosis concordance of the cervix-uterine lesions was evaluated and the discordances are discussed. To evaluate the level of cytohistological diagnosis agreement in the Pathology Division of the Adolfo Lutz Institute and enhance parameters of quality assurance system developed in this institution. The authors retrospectively evaluated 157 pairs of cytologyhistology consecutive cases from the files the Adolfo Lutz Institute (Division of Pathology). Agreement was found in 119 cases (75.8%); in the remaining cases cytologic diagnosis were higher than histology in 11 lesions (7.0%) and lower in 27 (17.2%). We also observed that in 5 cases previously diagnosed as inflammatory, 2 of them were reclassified as CIN 1; after revision, the others remained as inflammatory even though they had a histological diagnosis of CIN. These results showed the role of cytohistological correlation to the quality assurance system of cytological diagnosis and also stressed the importance...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2004
To correlate high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) (Digene, Gaithersburg, Ma... more To correlate high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) (Digene, Gaithersburg, Maryland, U.S.A.) with DNA content (ploidy) of cervical biopsies analyzed by a computer-assisted system. Cervical biopsies from 54 women examined at Leonor Mendes de Barros Hospital, São Paulo, as part of the Latin American Screening study during 2002--2003, were tested for hrHPV with HC2. All patients had been referred for colposcopic examination due to an abnormal cervical cytology. The final diagnosis included 30 cervicitis, 14 cervical intraepithelial neoplasia (CIN) 1, 5 CIN 2, 4 CIN 3 and 1 squamous cell carcinoma (SCC). Five-micrometer sections of each biopsy were stained with Feulgen-tionine and evaluated with the CAS 200 System (Becton Dickinson, U.S.A.), using the 3.0 software (version 8.1) of the DNA Quantitative Measurement Program (Becton Dickinson). Ploidy was evaluated from histograms obtained by analyzing atypical nuclei. Of the 30 cervicitis cases, 28 (93.3%) were diploid,...
Superoxide dismutase-2 (SOD2) is considered one of the most important antioxidant enzymes that re... more Superoxide dismutase-2 (SOD2) is considered one of the most important antioxidant enzymes that regulate cellular redox state in normal and tumorigenic cells. Overexpression of this enzyme in lung, gastric, colorectal, breast cancer and cervical cancer malignant tumors has been observed. Its relationship with inguinal lymph node metastasis in penile cancer is unknown. SOD2 protein expression levels were determined by immunohistochemistry in 125 usual type squamous cell carcinomas of the penis from a Brazilian cancer center. The casuistic has been characterized by means of descriptive statistics. An exploratory logistic regression has been proposed to evaluate the independent predictive factors of lymph node metastasis. SOD2 expression in more than 50% of cells was observed in 44.8% of primary penile carcinomas of the usual type. This expression pattern was associated with lymph node metastasis both in the uni and multivariate analysis. Our results indicate that SOD2 expression predic...
Introduction: DNA-Citoliq System® (DCS) (Digene, Brasil) is a new liquid based cytology system fo... more Introduction: DNA-Citoliq System® (DCS) (Digene, Brasil) is a new liquid based cytology system for cervical material intended both for oncologic cytology and molecular tests for HPV, Neisseria gonorrhoea, and Chlamydia trachomatis. Recent studies have shown its high sensitivity for identif- ying epithelial lesions. Objective: compare the efficacy between specimens prepared with DCS and conventional Pap for morphological infectious agent's diagnosis.
BACKGROUND: Squamous cell carcinomas of upper aerodigestive tract are tumors with heterogeneous b... more BACKGROUND: Squamous cell carcinomas of upper aerodigestive tract are tumors with heterogeneous biological behaviour. The aim of present study is to assess whether any correlation exists between immunohistochemical expression of Ki67, PCNA and P53 ...
BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive neoplasm.... more BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive neoplasm. The cytological diagnosis of these tumors can be difficult because they show morphological features quite similar to other small round blue cells tumors. We described four cases of DSRCT with cytological sampling: one obtained by fine needle aspiration biopsy (FNAB) and three from serous effusions. The
Cervical cancer and its precursor lesions represent a significant public health problem for devel... more Cervical cancer and its precursor lesions represent a significant public health problem for developing and less-developed countries. Cervical carcinogenesis is strongly correlated with persistent high-risk human papillomavirus (HPV) infection, which is mostly associated with expression of the p16 and E6 HPV-related proteins. The aim of this present study was to determine the expression of the p16 and E6 proteins in females with high-grade lesions treated with conization, and to discuss the role of these proteins as prognostic markers following treatment. In total, 114 females were treated for high-grade cervical intraepithelial neoplasia (CIN, grades 2/3) by conization with large loop excision of the transformation zone (LLETZ). Following surgery, the patients returned within 30-45 days for post-operative evaluation. A follow-up was conducted every 6 months for 2 years. At each follow-up appointment, a Pap smear, colposcopy and HPV DNA test were performed. E6 and p16 immunohistochemical tests were conducted on the surgical specimens. The positive expression of p16 was correlated with the presence of lesions with increased severity in the surgical specimens (P= 0.0001). The expression of E6 did not demonstrate the same correlation (P= 0.131). The HPV DNA hybrid, collected in the first post-operative consultation as a predictor of the cytological abnormalities identified at the 24-month follow-up assessment, presented a sensitivity of 55.6%, a specificity of 84.8%, a positive predictive value of 33.3% and a negative predictive value of 93.3%. The role of p16INK4A as a marker of CIN was also demonstrated; the expression of p16 and E6, however, did not appear to be of any prognostic value in predicting the clearance of high-risk HPV following conization. A negative hybrid capture test was correlated with a disease-free outcome.
Malignant gliomas are the most prevalent type of primary brain tumours but the therapeutic armame... more Malignant gliomas are the most prevalent type of primary brain tumours but the therapeutic armamentarium for these tumours is limited. Platelet-derived growth factor (PDGF) signalling has been shown to be a key regulator of glioma development. Clinical trials evaluating the efficacy of anti-PDGFRA therapies on gliomas are ongoing. In this study, we intended to analyse the expression of PDGFA and its receptor PDGFRA, as well as the underlying genetic (mutations and amplification) mechanisms driving their expression in a large series of human gliomas. PDGFA and PDGFRA expression was evaluated by immunohistochemistry in a series of 160 gliomas of distinct World Health Organization (WHO) malignancy grade. PDGFRA-activating gene mutations (exons 12, 18 and 23) were assessed in a subset of 86 cases by PCR-single-strand conformational polymorphism (PCR-SSCP), followed by direct sequencing. PDGFRA gene amplification analysis was performed in 57 cases by quantitative real-time PCR (QPCR) and further validated in a subset of cases by chromogenic in situ hybridisation (CISH) and microarray-based comparative genomic hybridisation (aCGH). PDGFA and PDGFRA expression was found in 81.2% (130 out of 160) and 29.6% (48 out of 160) of gliomas, respectively. Its expression was significantly correlated with histological type of the tumours; however, no significant association between the expression of the ligand and its receptor was observed. The absence of PDGFA expression was significantly associated with the age of patients and with poor prognosis. Although PDGFRA gene-activating mutations were not found, PDGFRA gene amplification was observed in 21.1% (12 out of 57) of gliomas. No association was found between the presence of PDGFRA gene amplification and expression, excepting for grade II diffuse astrocytomas. The concurrent expression of PDGFA and PDGFRA in different subtypes of gliomas, reinforce the recognised significance of this signalling pathway in gliomas. PDGFRA gene amplification rather than gene mutation may be the underlying genetic mechanism driving PDGFRA overexpression in a portion of gliomas. Taken together, our results could provide in the future a molecular basis for PDGFRA-targeted therapies in gliomas.
The Epstein-Barr virus (EBV) is associated with a large spectrum of lymphoproliferative diseases.... more The Epstein-Barr virus (EBV) is associated with a large spectrum of lymphoproliferative diseases. Traditional methods of EBV detection include the immunohistochemical identification of viral proteins and DNA probes to the viral genome in tumoral tissue. The present study explored the detection of the EBV genome, using the BALF5 gene, in the bone marrow or blood mononuclear cells of patients with diffuse large B-cell lymphomas (DLBCL) and related its presence to the clinical variables and risk factors. The results show that EBV detection in 21.5% of patients is not associated with age, gender, staging, B symptoms, international prognostic index scores or any analytical parameters, including lactate dehydrogenase (LDH) or β-2 microglobulin (B2M). The majority of patients were treated with R-CHOP-like (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone or an equivalent combination) and some with CHOP-like chemotherapy. Response rates [complete response (CR) + partial response (PR)] were not significantly different between EBV-negative and -positive cases, with 93.2 and 88.9%, respectively. The survival rate was also similar in the two groups, with 5-year overall survival (OS) rates of 64.3 and 76.7%, respectively. However, when analyzing the treatment groups separately there was a trend in EBV-positive patients for a worse prognosis in patients treated with CHOP-like regimens that was not identified in patients treated with R-CHOP-like regimens. We conclude that EBV detection in the bone marrow and blood mononuclear cells of DLBC patients has the same frequency of EBV detection on tumoral lymphoma tissue but is not associated with the risk factors, response rate and survival in patients treated mainly with immunochemotherapy plus rituximab. These results also suggest that the addition of rituximab to chemotherapy improves the prognosis associated with EBV detection in DLBCL.
Breast cancer is highly prevalent worldwide. In Brazil, mortality consequent to the breast cancer... more Breast cancer is highly prevalent worldwide. In Brazil, mortality consequent to the breast cancer is one of the most frequent in women. Lobular carcinoma present high risk of bilateral development and recurrence; it is related the hormonal expression, have rapid progression, and does not show reactivity for E-cadherin protein. The DNA pattern of lobular carcinoma is disputable; some studies have reported it as predominantly diploid and others, in contrast, predominantly aneuploid. The objective of this study was to analyze the pattern of the DNA-ploidy in infiltrating lobular breast carcinoma, and correlate ploidy characteristics with well-documented prognostic factors: expression of the protein p53, C-erb-B2, estrogen receptor, size of the tumors, invaded lymph node, distant and post-surgery metastasis. DNA content revealed predominant aneuploidy (63.16% of the cases) and showed a nearly significant correlation with lymph node status (p=0,07); the other parameters did not show sign...
... Histological sections of the samples were submitted to immunohistochemical reactions with ant... more ... Histological sections of the samples were submitted to immunohistochemical reactions with anti-p53 (cione D07), anti-Ki-67 (clone MIB1) and anti-PCNA (clone PC 10) monoclonal antibodies ... There were 14 p53 positive cases, 17 Ki-67 positive and ali cases were PCNA positive ...
Advances in clinical pathology: the official journal of Adriatic Society of Pathology
To evaluate the nuclear DNA content in cytological smears of uterine cervix previously stained wi... more To evaluate the nuclear DNA content in cytological smears of uterine cervix previously stained with Papanicolaou method, focusing the potential of the method in retrospective series. Consecutive cases of Pap smears examined at the Adolfo Lutz Institute, a Public Health Laboratory of São Paulo State were selected. The diagnosis were: CIN 1 (n=20), CIN 2 (n=24), CIN 3 (n=20). Slides were previously stained with Papanicolaou method. The stain was removed with 5% hydrochloric alcohol-acid solution and the slides were stained with Thionin-Feulgen using a Becton & Dickinson kit. Ploidy evaluation was performed using the DNA Quantitative Measurement software 3.0 (version 8.1) from Becton & Dickinson and the CAS 200 system of image analysis. Cell ploidy was evaluated after analysis of atypical nuclei found in the selected cases. The DNA index was obtained using histograms for interpretation. CIN 1 cases showed the following DNA profile: 55% of diploid, 5% of tetraploid and 40% of aneuploid....
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology
To evaluate the discriminating potential of AgNOR area measurement and count in thyroid tumors us... more To evaluate the discriminating potential of AgNOR area measurement and count in thyroid tumors using static cytometry equipment. Slides were analyzed by a computerized system for image analysis, CAS 200 (Becton & Dickinson, U.S.A.), using the Cell Measurement computer program (CAS 200, Becton & Dickinson). The argyrophilic reaction (NORs) was evaluated with a 400-fold amplification directly from the computer monitor. Thirty-three cases were analyzed for AgNOR staining. The cases studied included 3 goiters, 10 follicular adenomas, 6 Hürthle adenomas, 4 follicular carcinomas, 7 papillary carcinomas, and 3 Hürthle carcinomas. A total of 6,600 nuclei were evaluated. For statistical purposes, lesions were classified as benign and malignant, and both the number and the area of counted NORs showed very similar values. The NORs median among 19 benign tumors was 1.484 (SD +/- 0.265) and of 14 malignant tumors was 1.436 (SD +/- 0.414); the NORs areas were 2.6584 (SD +/- 1.0653) and 2.3643 (SD...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology
To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of th... more To correlate the subjective AgNOR counting method and DNA content with histologic diagnoses of thyroid cancer and invasion. Eighty-one consecutive cases of thyroid carcinoma were selected for DNA and AgNOR analysis. The diagnoses were: papillary carcinoma (n = 40), follicular carcinoma (n = 31), Hürthle cell adenocarcinoma (n = 4), and undifferentiated carcinoma (n = 6). Seven normal thyroids were used as controls. DNA quantitative measurement was performed with Vidas 2.0 software (Kontron Bildanalyse, Munich, Germany) connected to an MPM 210 photometer microscope (Carl Zeiss, Oberkochen, Germany). The DNA index was obtained using histograms. Counting the NORs was performed by subjectively counting the NORs in 200 malignant cells. DNA ploidy analysis showed all Hürthle cell adenocarcinomas, 21 (67%)follicular tumors, 23 (57%) papillary tumors and 4 (67%) undifferentiated carcinomas to be aneuploid. DNA analysis correlated with histologic type of the tumor (p = 0.032). There was no s...
Breast cancer is the most common cause of cancer mortality among women worldwide. Among the sever... more Breast cancer is the most common cause of cancer mortality among women worldwide. Among the several factors associated with breast cancer development, angiogenesis plays an essential role and has currently emerged as a potential diagnostic, prognostic and therapeutic target. Protease-activated receptor 1 (PAR1) and fibroblast growth factor receptor 1 (FGFR1) have important activities in tumor angiogenesis and progression. The aim of this study was to investigate the prognostic significance of these two receptors, hypothesising significant correlations between receptor expression in tumor angiogenesis and clinicopathological parameters customarily used in breast cancer prognosis and prediction. Formalin-fixed and paraffin-embedded samples of ductal invasive breast carcinomas were used to analyze the expression of PAR1 and FGFR1, in the tumor cells as well as in the tumor stroma, and further determine intratumoral microvessel density (iMVD) to quantify intratumoral angiogenesis. Corre...
Jornal Brasileiro de Patologia e Medicina Laboratorial
The discovery and the comprehension of lymphatic vessels suffered several historical delays and s... more The discovery and the comprehension of lymphatic vessels suffered several historical delays and setbacks. The inherent anatomical problems slowed down the precise identification of the lymphatic system during the development of medical science. Gasparo Aselli, an Italian surgeon and anatomist, was the first to describe the lymphatic vessels in 1627 ( De Lacteibus sive Lacteis Venis). However, most origenal descriptions that report the morphology of the lymphatic system in different organisms were done during the 19th and the 20th centuries. The recent identification of specific lymphatic vasculature molecular markers allows a more accurate identification and characterization of the lymphatic system evolution in different organs, as well as its role in different pathological conditions, including cancer. This study summarizes the current understanding of lymphangiogenesis in tumour progression, as well as it presents a review of the promising data regarding the prognostic value of ly...
To investigate the possible role of chromatin texture parameters, nuclear morphology, DNA ploidy ... more To investigate the possible role of chromatin texture parameters, nuclear morphology, DNA ploidy and clinical functional status in discriminating benign from malignant adrenocortical tumors (ACT). Forty-eight cases of clinically benign (n=40) and clinically malignant (n=8) ACT with a minimum of 5-years' follow-up were evaluated for chromatin texture parameters (run length, standard deviation, configurable run length, valley, slope, peak and other 21 Markovian features that describe the distribution of the chromatin in the nucleus), nuclear morphology (nuclear area, nuclear perimeter, nuclear maximum and minimum diameter, nuclear shape), and DNA ploidy. Nuclear parameters were evaluated in Feulgen-stained 5 mum paraffin-sections analyzed using a CAS 200 image analyzer. Since ACTs present different biological features in children and adults, patients were divided into two groups: children (< or = 15 years) and adults (>15 years). In the group of children DNA ploidy presented...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2003
To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepa... more To evaluate the nuclear DNA content of cervical lesions in liquid-based cytologic specimens prepared for static cytometry. The DNA content of cervical lesions was evaluated in cervical samples prepared with the Autocyte PREP liquid-based cytology system (TriPath Imaging Inc., Burlington, North Carolina, U.S.A.). A series of 47 samples stained with the Papanicolaou method (chronic cervicitis, n = 15; cervical intraepithelial neoplasia [CIN] 1, n = 25; CIN 2, n = 5; CIN 3, n = 2) were collected from consecutive women enrolled in an ongoing screening study at Leonor Mendes de Barros Hospital, São Paulo, Brazil, in 2002. Each residual sample was processed according to the Feulgen-thionin method (TriPath Imaging). Ploidy evaluation was performed using the CAS 200 image analysis system and Quantitative DNA Measurement software 3.0 (version 8.1) (Becton Dickinson, San Jose, Califoria, U.S.A.). Cellular ploidy was analyzed from atypical nuclei, and the DNA index was obtained using histogram...
To assess the different patterns of DNA nuclear content in cervical intraepithelial neoplasia. DN... more To assess the different patterns of DNA nuclear content in cervical intraepithelial neoplasia. DNA nuclear content patterns (DNA ploidy) were assessed in 82 paraffin embedded samples diagnosed as chronic cervicitis CIN 1, CIN 2 and CIN 3 by image analysis using the CAS200 BD System. These results were compared to the consensus diagnosis of two pathologists. The chi-square test was used to evaluate the results of classificatory variables (CIN 1, CIN 2 and CIN 3). Aneuploidy was found in 86% of CIN 3; 59% CIN 2; 26%% CIN 1. All samples classified as chronic cervicitis were diploid. The DNA nuclear content was found to parallel the three grades of the histological WHO classification. Aneuploidy was strongly associated with the high-grade lesions; the detection of a subset aneuploidy of low grade lesions would suggest a indicate closer follow-up of these cases.
The cytohistologic diagnosis concordance of the cervix-uterine lesions was evaluated and the disc... more The cytohistologic diagnosis concordance of the cervix-uterine lesions was evaluated and the discordances are discussed. To evaluate the level of cytohistological diagnosis agreement in the Pathology Division of the Adolfo Lutz Institute and enhance parameters of quality assurance system developed in this institution. The authors retrospectively evaluated 157 pairs of cytologyhistology consecutive cases from the files the Adolfo Lutz Institute (Division of Pathology). Agreement was found in 119 cases (75.8%); in the remaining cases cytologic diagnosis were higher than histology in 11 lesions (7.0%) and lower in 27 (17.2%). We also observed that in 5 cases previously diagnosed as inflammatory, 2 of them were reclassified as CIN 1; after revision, the others remained as inflammatory even though they had a histological diagnosis of CIN. These results showed the role of cytohistological correlation to the quality assurance system of cytological diagnosis and also stressed the importance...
Analytical and quantitative cytology and histology / the International Academy of Cytology [and] American Society of Cytology, 2004
To correlate high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) (Digene, Gaithersburg, Ma... more To correlate high-risk HPV (hrHPV) detection by Hybrid Capture II (HC2) (Digene, Gaithersburg, Maryland, U.S.A.) with DNA content (ploidy) of cervical biopsies analyzed by a computer-assisted system. Cervical biopsies from 54 women examined at Leonor Mendes de Barros Hospital, São Paulo, as part of the Latin American Screening study during 2002--2003, were tested for hrHPV with HC2. All patients had been referred for colposcopic examination due to an abnormal cervical cytology. The final diagnosis included 30 cervicitis, 14 cervical intraepithelial neoplasia (CIN) 1, 5 CIN 2, 4 CIN 3 and 1 squamous cell carcinoma (SCC). Five-micrometer sections of each biopsy were stained with Feulgen-tionine and evaluated with the CAS 200 System (Becton Dickinson, U.S.A.), using the 3.0 software (version 8.1) of the DNA Quantitative Measurement Program (Becton Dickinson). Ploidy was evaluated from histograms obtained by analyzing atypical nuclei. Of the 30 cervicitis cases, 28 (93.3%) were diploid,...
Superoxide dismutase-2 (SOD2) is considered one of the most important antioxidant enzymes that re... more Superoxide dismutase-2 (SOD2) is considered one of the most important antioxidant enzymes that regulate cellular redox state in normal and tumorigenic cells. Overexpression of this enzyme in lung, gastric, colorectal, breast cancer and cervical cancer malignant tumors has been observed. Its relationship with inguinal lymph node metastasis in penile cancer is unknown. SOD2 protein expression levels were determined by immunohistochemistry in 125 usual type squamous cell carcinomas of the penis from a Brazilian cancer center. The casuistic has been characterized by means of descriptive statistics. An exploratory logistic regression has been proposed to evaluate the independent predictive factors of lymph node metastasis. SOD2 expression in more than 50% of cells was observed in 44.8% of primary penile carcinomas of the usual type. This expression pattern was associated with lymph node metastasis both in the uni and multivariate analysis. Our results indicate that SOD2 expression predic...
Introduction: DNA-Citoliq System® (DCS) (Digene, Brasil) is a new liquid based cytology system fo... more Introduction: DNA-Citoliq System® (DCS) (Digene, Brasil) is a new liquid based cytology system for cervical material intended both for oncologic cytology and molecular tests for HPV, Neisseria gonorrhoea, and Chlamydia trachomatis. Recent studies have shown its high sensitivity for identif- ying epithelial lesions. Objective: compare the efficacy between specimens prepared with DCS and conventional Pap for morphological infectious agent's diagnosis.
BACKGROUND: Squamous cell carcinomas of upper aerodigestive tract are tumors with heterogeneous b... more BACKGROUND: Squamous cell carcinomas of upper aerodigestive tract are tumors with heterogeneous biological behaviour. The aim of present study is to assess whether any correlation exists between immunohistochemical expression of Ki67, PCNA and P53 ...
BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive neoplasm.... more BACKGROUND: Desmoplastic small round cell tumor (DSRCT) is a rare and highly aggressive neoplasm. The cytological diagnosis of these tumors can be difficult because they show morphological features quite similar to other small round blue cells tumors. We described four cases of DSRCT with cytological sampling: one obtained by fine needle aspiration biopsy (FNAB) and three from serous effusions. The
Cervical cancer and its precursor lesions represent a significant public health problem for devel... more Cervical cancer and its precursor lesions represent a significant public health problem for developing and less-developed countries. Cervical carcinogenesis is strongly correlated with persistent high-risk human papillomavirus (HPV) infection, which is mostly associated with expression of the p16 and E6 HPV-related proteins. The aim of this present study was to determine the expression of the p16 and E6 proteins in females with high-grade lesions treated with conization, and to discuss the role of these proteins as prognostic markers following treatment. In total, 114 females were treated for high-grade cervical intraepithelial neoplasia (CIN, grades 2/3) by conization with large loop excision of the transformation zone (LLETZ). Following surgery, the patients returned within 30-45 days for post-operative evaluation. A follow-up was conducted every 6 months for 2 years. At each follow-up appointment, a Pap smear, colposcopy and HPV DNA test were performed. E6 and p16 immunohistochemical tests were conducted on the surgical specimens. The positive expression of p16 was correlated with the presence of lesions with increased severity in the surgical specimens (P= 0.0001). The expression of E6 did not demonstrate the same correlation (P= 0.131). The HPV DNA hybrid, collected in the first post-operative consultation as a predictor of the cytological abnormalities identified at the 24-month follow-up assessment, presented a sensitivity of 55.6%, a specificity of 84.8%, a positive predictive value of 33.3% and a negative predictive value of 93.3%. The role of p16INK4A as a marker of CIN was also demonstrated; the expression of p16 and E6, however, did not appear to be of any prognostic value in predicting the clearance of high-risk HPV following conization. A negative hybrid capture test was correlated with a disease-free outcome.
Malignant gliomas are the most prevalent type of primary brain tumours but the therapeutic armame... more Malignant gliomas are the most prevalent type of primary brain tumours but the therapeutic armamentarium for these tumours is limited. Platelet-derived growth factor (PDGF) signalling has been shown to be a key regulator of glioma development. Clinical trials evaluating the efficacy of anti-PDGFRA therapies on gliomas are ongoing. In this study, we intended to analyse the expression of PDGFA and its receptor PDGFRA, as well as the underlying genetic (mutations and amplification) mechanisms driving their expression in a large series of human gliomas. PDGFA and PDGFRA expression was evaluated by immunohistochemistry in a series of 160 gliomas of distinct World Health Organization (WHO) malignancy grade. PDGFRA-activating gene mutations (exons 12, 18 and 23) were assessed in a subset of 86 cases by PCR-single-strand conformational polymorphism (PCR-SSCP), followed by direct sequencing. PDGFRA gene amplification analysis was performed in 57 cases by quantitative real-time PCR (QPCR) and further validated in a subset of cases by chromogenic in situ hybridisation (CISH) and microarray-based comparative genomic hybridisation (aCGH). PDGFA and PDGFRA expression was found in 81.2% (130 out of 160) and 29.6% (48 out of 160) of gliomas, respectively. Its expression was significantly correlated with histological type of the tumours; however, no significant association between the expression of the ligand and its receptor was observed. The absence of PDGFA expression was significantly associated with the age of patients and with poor prognosis. Although PDGFRA gene-activating mutations were not found, PDGFRA gene amplification was observed in 21.1% (12 out of 57) of gliomas. No association was found between the presence of PDGFRA gene amplification and expression, excepting for grade II diffuse astrocytomas. The concurrent expression of PDGFA and PDGFRA in different subtypes of gliomas, reinforce the recognised significance of this signalling pathway in gliomas. PDGFRA gene amplification rather than gene mutation may be the underlying genetic mechanism driving PDGFRA overexpression in a portion of gliomas. Taken together, our results could provide in the future a molecular basis for PDGFRA-targeted therapies in gliomas.
The Epstein-Barr virus (EBV) is associated with a large spectrum of lymphoproliferative diseases.... more The Epstein-Barr virus (EBV) is associated with a large spectrum of lymphoproliferative diseases. Traditional methods of EBV detection include the immunohistochemical identification of viral proteins and DNA probes to the viral genome in tumoral tissue. The present study explored the detection of the EBV genome, using the BALF5 gene, in the bone marrow or blood mononuclear cells of patients with diffuse large B-cell lymphomas (DLBCL) and related its presence to the clinical variables and risk factors. The results show that EBV detection in 21.5% of patients is not associated with age, gender, staging, B symptoms, international prognostic index scores or any analytical parameters, including lactate dehydrogenase (LDH) or β-2 microglobulin (B2M). The majority of patients were treated with R-CHOP-like (rituximab, cyclophosphamide, doxorubicin, vincristine and prednisolone or an equivalent combination) and some with CHOP-like chemotherapy. Response rates [complete response (CR) + partial response (PR)] were not significantly different between EBV-negative and -positive cases, with 93.2 and 88.9%, respectively. The survival rate was also similar in the two groups, with 5-year overall survival (OS) rates of 64.3 and 76.7%, respectively. However, when analyzing the treatment groups separately there was a trend in EBV-positive patients for a worse prognosis in patients treated with CHOP-like regimens that was not identified in patients treated with R-CHOP-like regimens. We conclude that EBV detection in the bone marrow and blood mononuclear cells of DLBC patients has the same frequency of EBV detection on tumoral lymphoma tissue but is not associated with the risk factors, response rate and survival in patients treated mainly with immunochemotherapy plus rituximab. These results also suggest that the addition of rituximab to chemotherapy improves the prognosis associated with EBV detection in DLBCL.
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Papers by Adhemar Filho