Immunoexpression of BAP1, ROS1, and ALK in Spitzoid Melanocytic Tumors.

BACKGROUND: Spitzoid tumors are a heterogeneous group of melanocytic neoplasms that frequently imposes diagnostic difficulties. Lately, several advances in molecular biology afforded significant discoveries on the pathogenesis of these tumors. BAP1 (BRCA-1 associated protein-1) inactivation and anomalous expression of kinase translocation-related proteins are among the main criteria launched by new classification proposals. Our aim was to systematically assess the immunoexpression of BAP1, ROS1 (receptor tyrosine kinase c-Ros oncogene 1), and ALK (anaplastic lymphoma receptor tyrosine kinase) proteins in an unpublished series of spitzoid tumors. METHODS: Retrospective study based on 47 formalin-fixed paraffin-embedded tissue samples from 3 different institutions. BAP1, ROS1, and ALK immunostains were performed in all cases. We included 27 Spitz tumors without significant abnormality, 15 atypical spitzoid tumors, and 5 spitzoid melanomas. RESULTS: We observed loss of BAP1 nuclear immunolabeling in 4.3% of evaluable cases (2/46), both of them atypical spitzoid tumors. The proportional frequency of BAP1-inactivated cases among atypical spitzoid tumors was 14.2% (2/14). No immunoexpression of ROS1 or ALK was found. CONCLUSIONS: Our study revealed 2 additional BAP1-inactived cases and described its respective frequency. The absence of anomalous expression of translocation-related proteins ALK and ROS1 in this series, composed predominantly of low-grade/low-risk tumors, indicates that translocated spitzoid lesions may not be as prevalent as initially suggested, at least in some populations. Furthermore, our findings encourage additional investigation on unequal occurrence of such immunomarkers among different diagnostic categories of spitzoid neoplasms.

Expression of tyrosine kinase receptor AXL is associated with worse outcome of metastatic renal cell carcinomas treated with sunitinib.

BACKGROUND: Renal cell carcinoma (RCC) represents 2%-3% of all cancers of the Western countries. Currently, sunitinib, a receptor tyrosine kinase inhibitor, particularly of PDGF and VEGF receptors, is the first-line therapy for metastatic RCC (mRCC), with significant improvement in clinical outcome. However, there is a lack of predictive biomarkers of sunitinib response. Recently, others and our group suggested that the receptor tyrosine kinase AXL may modify the response to sunitinib. OBJECTIVE: To study the expression of AXL in a series patients with of mRCC treated with sunitinib and to correlate it with patient's clinic-pathological features and therapeutic response. MATERIAL AND METHODS: Sixty-four patients with mRCC (51 clear cell carcinomas (CCCs) and 13 non-CCCs) were evaluated for AXL expression by immunohistochemistry in the primary tumor. RESULTS: AXL positivity was observed in 47% (30/64) of cases, namely in 43% (22/51) of CCCs and 61% (8/13) of non-CCC. Considering only the clear cell subtype, the univariate analysis showed that AXL expression was statistically associated with a poor prognosis, with a median overall survival of 13 months vs. 43 months in patients with negative AXL. In this subtype, along with the AXL positivity, other prognostic factors were absence of nephrectomy, Karnofsky performance status, more than 1 site of metastasis and liver metastasis. Moreover, AXL expression was associated with shorter progression to sunitinib. Overall, the multivariate survival analysis showed that absence of nephrectomy (HR = 4.85, P = 0.001), more than 1 site of metastasis (HR = 2.99, P = 0.002), bone metastasis (HR = 2.95, P = 0.001), together with AXL expression (HR = 2.01, P = 0.048) were independent poor prognostic factor in patients with mRCC. CONCLUSION: AXL expression was associated with worse clinical outcome and may be an important prognostic biomarker in sunitinib-treated patients with metastatic renal cell carcinoma.

An integrated tool for determining the primary origin site of metastatic tumours.

AIMS: Cancers of unknown primary sites account for 3%-5% of all malignant neoplasms. Current diagnostic workflows based on immunohistochemistry and imaging tests have low accuracy and are highly subjective. We aim to develop and validate a gene-expression classifier to identify potential primary sites for metastatic cancers more accurately. METHODS: We built the largest Reference Database (RefDB) reported to date, composed of microarray data from 4429 known tumour samples obtained from 100 different sources and divided into 25 cancer superclasses formed by 58 cancer subclass. Based on specific profiles generated by 95 genes, we developed a gene-expression classifier which was first trained and tested by a cross-validation. Then, we performed a double-blinded retrospective validation study using a real-time PCR-based assay on a set of 105 metastatic formalin-fixed, paraffin-embedded (FFPE) samples. A histopathological review performed by two independent pathologists served as a reference diagnosis. RESULTS: The gene-expression classifier correctly identified, by a cross-validation, 86.6% of the expected cancer superclasses of 4429 samples from the RefDB, with a specificity of 99.43%. Next, the performance of the algorithm for classifying the validation set of metastatic FFPE samples was 83.81%, with 99.04% specificity. The overall reproducibility of our gene-expression-classifier system was 97.22% of precision, with a coefficient of variation for inter-assays and intra-assays and intra-lots <4.1%. CONCLUSION: We developed a complete integrated workflow for the classification of metastatic tumour samples which may help on tumour primary site definition.

A rare case of TFE-related pigmented renal tumor with overlapping features between melanotic Xp11 translocation renal cancer and Xp11 renal cell carcinoma with melanotic features.

In recent years, an increasing number of TFE3 rearrangement-associated tumors with melanotic features have been reported as primary neoplasm in different anatomical sites, including the kidney. Melanotic Xp11 translocation renal cancer (MXTRC) and Xp11 renal cell carcinoma with melanotic features (XRCCM) have been proposed to be main categories for pigmented lesions in the microophthalmia-associated transcription factor (MiTF/TFE3) family of renal tumors that may show variable degrees of melanocytic differentiation. Herein we report a rare case of TFE3-related pigmented renal tumor showing unusual immunoexpression of cytokeratins (AE1/AE3) and renal cell carcinoma markers (RCC, CD10). Cathepsin-K and Vimentin were diffusely positive whereas melanocytic markers (HMB-45 and Melan-A) displayed weak and patchy expression. We found no labelling for PAX-8, muscle markers (desmin, smooth muscle actin, muscle-specific actin and caldesmon) and S-100. TFE3 fusion was confirmed by break-apart fluorescence in situ hybridization (FISH). This case corroborates previous evidence for overlap in the TFE3-associated cancer family and illustrates that it may not be possible to set a clear cutoff between epithelial (XRCCM) and mesenchymal (MXTRC) subgroups.

Expressão imuno-histoquímica das proteínas VEGF e HER-2 em biópsias de osteossarcoma / Immunohistochemical expression of VEGF and HER-2 proteins in osteosarcoma biopsies

Objetivos: Identificar a prevalência de HER-2 e do fator de crescimento do endotélio vascular (VEGF) em biópsias de osteossarcoma e correlacioná-los com possíveis fatores de prognóstico. Métodos: estudo retrospectivo realizado no Hospital de Câncer de Barretos-SP incluindo 27 biópsias de osteossarcoma imuno-histoquimicamente coradas para VEGF e HER-2. Características clínico-patológicas foram coletadas dos prontuários médicos dos pacientes para correlação com marcadores. Resultados: em 27 biópsias, quatro foram superexpressas para VEGF e três para HER-2. Dois terços dos pacientes eram não metastáticos. Quase todos pacientes com VEGF superexpresso apresentaram metástases. A superexpressão para HER-2 apresentou relação inversa à presença de metástases. Não houve associação significativa entre os marcadores e o prognóstico. Conclusão: identificamos baixa prevalência de VEGF e HER-2 na amostra. Não houve associação significativa entre superexpressão dos marcadores e características clínico-patológicas. A ampliação da amostra e do tempo de seguimento, além do emprego de novas técnicas laboratoriais pode determinar a real expressão de VEGF e HER-2 e seu papel em osteossarcomas. Nível de Evidência III, Estudo de Caso-controle.

Objectives: To identify the prevalence of erbB-2 and vascular endothelial growth factor (VEGF) in osteosarcoma biopsies and to correlate them with possible prognosis factors. Methods: retrospective study conducted at the Hospital do Câncer de Barretos-SP including 27 osteosarcoma biopsies immunohistochemically stained for VEGF and erbB-2. The pathological characteristics were collected from medical records of patients to correlate with markers. Results: In 27 biopsies, four overexpressed VEGF and three overexpressed erbB-2. Two thirds of patients had no metastases. Almost all patients with overexpression of VEGF showed metastases. Overexpression of erbB-2 was inversely related to the presence of metastases. There was no significant association between markers and prognosis. Conclusion: We identified a low prevalence of erbB-2 and VEGF in the sample. There was no significant association between overexpression of markers and pathological features. A larger sample and a longer follow-up, in addition to using new laboratory techniques can determine the real expression of VEGF and erbB-2 and its role in osteosarcoma. Level of Evidence III, Case-Control Study.

Malignant fat-forming solitary fibrous tumor (lipomatous hemangiopericytoma) in the neck: Imaging and histopathological findings of a case.

Fat-forming solitary fibrous tumor (SFT) is a rare variant of solitary fibrous tumor, a mesenchymal fibroblastic neoplasia with a particular branching hypervascular pattern. This tumor is usually classified as benign and only very few fat-forming SFTs with malignant histologic features have been reported. We report a histologically malignant fat-forming solitary fibrous tumor in a 61-year-old man, located in his neck. Ultrasonography examination was first performed showing a heterogeneous lesion, predominantly hyperechoic, with sound beam attenuation, containing two hypoechoic solid nodules. Magnetic resonance imaging and computed tomography examinations demonstrated a heterogeneous and predominantly adipose mass, containing post contrast enhancing solid nodules and thin septations. Treatment consisted of total removal of the lesion. Histologically, the tumor showed hypercellularity, numerous mitoses and cytological atypia, fulfilling the criteria for malignancy. The patient had no metastasis. This rare tumor may be confused with other fat-containing lesions on imaging examinations, mainly liposarcoma.

Immunohistochemical expression of vegf and her-2 proteins in osteosarcoma biopsies.

OBJECTIVES: To identify the prevalence of erbB-2 and vascular endothelial growth factor (VEGF) in osteosarcoma biopsies and to correlate them with possible prognosis factors. METHODS: Retrospective study conducted at the Hospital do Câncer de Barretos-SP including 27 osteosarcoma biopsies immunohistochemically stained for VEGF and erbB-2. The pathological characteristics were collected from medical records of patients to correlate with markers. RESULTS: In 27 biopsies, four overexpressed VEGF and three overexpressed erbB-2. Two thirds of patients had no metastases. Almost all patients with overexpression of VEGF showed metastases. Overexpression of erbB-2 was inversely related to the presence of metastases. There was no significant association between markers and prognosis. CONCLUSION: We identified a low prevalence of erbB-2 and VEGF in the sample. There was no significant association between overexpression of markers and pathological features. A larger sample and a longer follow-up, in addition to using new laboratory techniques can determine the real expression of VEGF and erbB-2 and its role in osteosarcoma . Level of Evidence III, Case-Control Study.

The interference of cold ischemia time in the quality of total RNA from frozen tumor samples.

Tumor Banks were created to organize the collection, storage and distribution of biological samples from oncological patients, facilitating its use in cancer research. To ensure the quality of the samples from our bank, we implemented standard operating procedures international. In order to evaluate the influence of cold ischemia time (time between surgical removal of the specimen and the snap freezing of the sample) on the quality of the samples (evaluated by measurement integrity of their RNA), collected during 10 months two tumor samples from each donor, one with up to 30 min of cold ischemia and other with exact 45 min, totaling 100 different donors and 200 samples, 40 from each of the following organs: breast, thyroid, stomach, lung and colorectum. We extracted total RNA from the samples and with the aid of a Bioanalyser, evaluate their quality, comparing it with cold ischemia times in different organs. Among the samples up to 30 min and the samples with exact 45 min, we respectively found 63 (64.3 %) and 36 (36 %) with intact RNA, 11 (11.2 %) and 17 (17 %) partially degraded and 24 (24.5 %) and 47 (47 %) degraded (p < 0.001). Thyroid and colorectal samples were more sensitive to variations in cold ischemia time (p = 0.006 and p = 0.03, respectively). Stomach and lungs were less sensitive (p = 0.919 and p = 0.384, respectively). We concluded that the cold ischemia time up to 30 min was more efficient to maintain the integrity of RNA in most samples, and that RNA degradation varied according to the different topographies.

Avaliação do impacto da implantação do controle de qualidade em um banco de amostras teciduais criopreservadas / Evaluating the impact of implementation of quality control in a bank of cryopreserved tissue samples

Bancos de tumores foram criados para organizar a coleta, arm azenamento e distribuição de amostras biológicas de pacientes oncológicos, favorecendo seu uso nas pesquis as sobre o cân cer. Amostras ade quadas devem ter RNA, DNA e proteínas de boa qualidade. RNA de boa qualidade deve estar íntegro e puro e DNA deve ter boa c oncentração e pur eza. Basea do em norm as in ternacionais, f oi elaborado e implantado um abrangente sistema de controle de qualidade no banco de tumores do Hospital de Câncer de Barretos, que para fins de estudo foi dividido em banco pré-controle de qu alidade (den ominado b anco pré) e em ban co pós- controle de qualidade (denominado banco pós). Objetivando comparar a qualidade das amostras n os dois bancos, atra vés d a extração d e R NA total e d e DNA (utilizando-se homogeneizador de tecidos e Kits), selecionou-se de forma aleatória 200 a mostras tumorais, distribuídas ig ualitariamente entre mama, co lorreto, estômago, pulmão e tireóide, sendo 100 do banco pré e 100 do banco pós. Para se avaliar a influência do tempo de isquemia fria (tempo entre a excisão do e spécime cirúrgico e o congelamento rápido da amostra armazenada) na qualidade do RNA total de amostras tumorais do banco pós, foram coletadas 200 amostras tumorais, distribuídas igualitariamente entre mama, co lorreto, estômago, pulmão e ti reóide, de 100 doadores diferentes, metade com o tempo de isquemia fria (TIF) de até 30 minutos e a o utra metade do mesmo espécime com TIF de 45 minutos. Extraiu-se RNA total dessas amostras (com maceração manual e Trizol) e comparou-se a sua qualidade, através do núm ero de i ntegridade do RNA (RIN), dentr o dos d ois intervalos de tempo e nas diferentes top ografias. Ao c omparar-se amostras com RIN acima de 7 (consideradas ideais para experimentos de microarray), do banco pré e do b anco pó s, for am enc ontrados 73 (73%) no p rimeiro e 87 (87%) no segundo (p=0,013). Ao comparar-se o intervalo de TIF de até 30 minutos com o de 45...

Tumor banks were created to or ganize the collection, storage and d istribution of biological samples of cancer pa tients, favoring it's use in cancer rese arches. Appropriate samples should have good quality of RNA, DNA and p roteins. RNA of good quality should be intact and pure and DNA should have good concentration and pu rity. Ba sed on international sta ndards, we elabo rated and imp lanted an comprehensive s ystem of qu ality control in the tu mor bank of Ba rretos Cancer Hospital, w hich was divided for st udy purposes i n pre bank quality control (denominated pre bank) and post bank qu ality control (denominated post bank). Aiming to compare the quality of the samples in two banks, through the extraction of total RNA and DNA (b y tissue homogenizer and Kits), we se lected 200 tumor samples in a random way, distributed equally among breast, colorectal, stomach, lung and thyroid, being 100 of the pre-bank and 100 of the post bank. To evaluate the influence o f cold ischem ia time (time b etween t he ex cision o f the su rgical specimen and the fast freezing of the stored sample) in the quality of total of RNA tumor sa mples of th e po st bank , we collected 2 00 t umor s amples, distrib uted equally among breast, colorectal, stom ach, lung and th yroid, fro m 100 different donors, half with the cold ischemia time (CIT) up to 30 minutes and the other ha lf of the sam e specimen with CIT exact ly 45 minutes. We ex tracted total RNA of these samples (with manual maceration and T rizol) and c ompared their qu ality, through the RNA integri ty number (RIN), ins ide tw o intervals of time a nd in different topographies. Comparing samples with RIN above 7 (considered ideals for microarray experiments), of the pre bank and of the post bank, we found 73 (73%) in the first and 87 (87%) in the second (p=0,013). Comparing the interval of CIT up to 30 m inutes with the ex actly 45 minutes, we found respectively 63 (64,3%)...

Necrotizing soft tissue infection of the breast: case report and literature review.

BACKGROUND: Necrotizing soft tissue infection (NSTI) is characterized by progressive infectious gangrene of the skin and subcutaneous tissue. Its treatment involves intensive care, broad-spectrum antibiotic therapy, and full debridement. METHODS: We present two cases of NSTI of the breast, adding these cases to the 14 described in the literature, reviewing the characteristics and evolution of all cases. CASE REPORT: On the fourth day after mastectomy, a 59-year-old woman with ulcerated breast cancer developed Type I NSTI caused by Pseudomonas aeruginosa, which had a favorable evolution after debridement and broad-spectrum antibiotics. The second patient was a 57-year-old woman submitted to a mastectomy and axillary dissection, who had recurrent seromas. On the 32nd post-operative day, after a seroma puncture, she developed Type II NSTI caused by ß-hemolytic streptococci. She developed sepsis and died on the tenth day after debridement, intensive care, and broad-spectrum antibiotics. The cases are the first description of breast NSTI after mammary seroma aspiration and the first report of this condition caused by P. aeruginosa. CONCLUSION: Necrotizing soft tissue infection is rare in breast tissue. It frequently is of Type II, occurring mainly after procedures in patients with breast cancer. The surgeon's participation in controlling the focus of the infection is of fundamental importance, and just as important are broad-spectrum antibiotic therapy and support measures, such as maintenance of volume, correction of electrolytic disorders, and treatment of sepsis and septic shock. Once the infection has been brought under control, skin grafting or soft tissue flaps can be considered. The mortality rate in breast NSTI is 18.7%, all deaths being in patients with the fulminant Type II form. Surgical oncologists need to be alert to the possibility of this rare condition.

Giant cell tumor of the sternum.

We report the case of a 74-year-old female patient diagnosed with a giant cell tumor of the sternum. The clinical and radiological presentation was indicative of a primary tumor of the sternum. The patient underwent complementary tests and surgery. The pathological examination confirmed the diagnosis. Commonly observed in the long bones of the appendicular skeleton, this type of tumor is characterized by its local aggressiveness and metastatic potential. We also review the literature on the topic.

Tumor de células gigantes do esterno / Giant cell tumor of the sternum

Relatamos o caso de uma paciente de 74 anos portadora de tumor de células gigantes em osso esterno. A apresentação clínica e radiológica indicava neoplasia primária de osso esterno. A paciente realizou exames complementares e cirurgia. O exame patológico confirmou o diagnóstico. Comumente observado em ossos longos no esqueleto apendicular, esse tipo de tumor caracteriza-se por sua agressividade local e pelo potencial metastático. Também fazemos uma revisão sobre o tema.

We report the case of a 74-year-old female patient diagnosed with a giant cell tumor of the sternum. The clinical and radiological presentation was indicative of a primary tumor of the sternum. The patient underwent complementary tests and surgery. The pathological examination confirmed the diagnosis. Commonly observed in the long bones of the appendicular skeleton, this type of tumor is characterized by its local aggressiveness and metastatic potential. We also review the literature on the topic.

Childhood carcinoid tumors: description of a case series in a Brazilian cancer center.

CONTEXT AND OBJECTIVE: Carcinoid tumors are very rare both in children and adults. About 85% of these tumors develop in the gastrointestinal tract. The objective of the present study was to describe our experience with children treated of carcinoid tumors, and investigate the frequency morphological findings and results. DESIGN AND SETTING: Report on case series, at the Department of Pediatrics of Centro de Tratamento e Pesquisa Hospital do Câncer, São Paulo. METHODS: This was a retrospective analysis of clinical pathological data and outcomes among children (< 18 years old) with carcinoid tumors admitted from January 1, 1990, to December 31, 2001. RESULTS: Nine patients (mean age 12.2 years) were included: six girls and three boys (2:1), all of them Caucasian. In eight cases (89%), the primary tumor site was the appendix and in one (11%) it was the left bronchus. For those with primary tumor in the appendix, the main complaint was abdominal pain, which led to appendectomy. Only one patient underwent right hemicolectomy due to tumor extension into the serosa. The patient with bronchial tumor underwent left pneumonectomy. All patients had localized disease and are alive and free of disease. They have had follow-ups lasting from 1 to 11 years (mean of 3.5 years). CONCLUSION: Although the majority of carcinoid tumors arise from the appendix, these tumors can also occur in other primary sites. Surgical resection at an early stage allows for good prognosis without the need for any adjuvant treatment.

Childhood carcinoid tumors: description of a case series in a Brazilian cancer center

CONTEXTO E OBJETIVO: Tumores carcinóides são bastante raros, tanto em crianças quanto em adultos. Cerca de 85% deles desenvolvem-se no trato gastrintestinal. O objetivo do presente estudo foi descrever a experiência de nossa instituição com crianças tratadas de tumores carcinóides, investigando sua freqüência, achados morfológicos e resultados. TIPO DE ESTUDO E LOCAL: Relato de série de casos, no Departmento de Pediatria do Centro de Tratamento e Pesquisa Hospital do Câncer, São Paulo, Brasil. MÉTODOS: Análise retrospectiva de dados clinicopatológicos e resultados de crianças menores de 18 anos de idade com tumores carcinóides, admitidas entre 1º de janeiro de 1990 e 31 de dezembro de 2001. RESULTADOS: O estudo inclui nove pacientes, todos brancos, com idade média de 12,2 anos, sendo seis meninas e três meninos (2:1). O apêndice foi o sítio primário em oito casos (89%) e o brônquio esquerdo em um caso (11%). Nos primeiros, o sintoma principal foi dor abdominal que levou a apendicectomia. Apenas um paciente foi submetido a hemicolectomia direita em razão da extensão do tumor na serosa. O paciente que tinha o tumor primário no brônquio foi submetido a pneumectomia à esquerda. Todos os pacientes tinham doença localizada e estão vivos sem doença, com acompanhamento de 1 a 11 anos (média de 3,5 anos). CONCLUSÃO: Embora a maioria dos tumores carcinóides tenha sua origem no apêndice, eles podem ocorrer em outros sítios. A ressecção cirúrgica em estágios iniciais permite um bom prognóstico sem necessidade de tratamento adjuvante.

[A large atypical meningioma in childhood: case report]. / Extenso meningioma atípico na infância: relato de caso.

Meningiomas are rare in children and a small number of cases has been published. They are tumors typically indolent, but sometimes they show an extreme aggressive behavior. The gold standard of treatment is surgery with total remove of tumor. If the surgery is not possible to do, the options of treatment are few. We describe a case of an atipical meningioma in a 3 years-old girl.

Extenso meningioma atípico na infância: relato de caso / A large atypical meningioma in childhood: case report

Meningiomas säo raros na criança e pequeno número de casos foi publicado até hoje. Eles säo tumores tipicamente indolentes, mas, podem apresentar comportamento mais agressivo na infância. A conduta ideal é a cirurgia com ressecçäo completa do tumor. Na impossibilidade da remoçäo cirúrgica completa do tumor, as opçöes de tratamento säo reduzidas. Nosso objetivo é descrever um caso de meningioma atípico na infância em menina de três anos, e sua evoluçäo