CCL21-CCR7 signaling promotes microglia/macrophage recruitment and chemotherapy resistance in glioblastoma.

Glioblastoma (GBM) is the most common and fatal primary tumor of the central nervous system (CNS) and current treatments have limited success. Chemokine signaling regulates both malignant cells and stromal cells of the tumor microenvironment (TME), constituting a potential therapeutic target against brain cancers. Here, we investigated the C-C chemokine receptor type 7 (CCR7) and the chemokine (C-C-motif) ligand 21 (CCL21) for their expression and function in human GBM and then assessed their therapeutic potential in preclinical mouse GBM models. In GBM patients, CCR7 expression positively associated with a poor survival. CCL21-CCR7 signaling was shown to regulate tumor cell migration and proliferation while also controlling tumor associated microglia/macrophage recruitment and VEGF-A production, thereby controlling vascular dysmorphia. Inhibition of CCL21-CCR7 signaling led to an increased sensitivity to temozolomide-induced tumor cell death. Collectively, our data indicate that drug targeting of CCL21-CCR7 signaling in tumor and TME cells is a therapeutic option against GBM.

The orthotopic xenotransplant of human glioblastoma successfully recapitulates glioblastoma-microenvironment interactions in a non-immunosuppressed mouse model.

BACKGROUND: Glioblastoma (GBM) is the most common primary brain tumor and the most aggressive glial tumor. This tumor is highly heterogeneous, angiogenic, and insensitive to radio- and chemotherapy. Here we have investigated the progression of GBM produced by the injection of human GBM cells into the brain parenchyma of immunocompetent mice. METHODS: Xenotransplanted animals were submitted to magnetic resonance imaging (MRI) and histopathological analyses. RESULTS: Our data show that two weeks after injection, the produced tumor presents histopathological characteristics recommended by World Health Organization for the diagnosis of GBM in humans. The tumor was able to produce reactive gliosis in the adjacent parenchyma, angiogenesis, an intense recruitment of macrophage and microglial cells, and presence of necrosis regions. Besides, MRI showed that tumor mass had enhanced contrast, suggesting a blood-brain barrier disruption. CONCLUSIONS: This study demonstrated that the xenografted tumor in mouse brain parenchyma develops in a very similar manner to those found in patients affected by GBM and can be used to better understand the biology of GBM as well as testing potential therapies.

Clotrimazole disrupts glycolysis in human breast cancer without affecting non-tumoral tissues.

Human breast cancer tissues, as well as normal tissues from the same patients, were treated with clotrimazole (CTZ) and have their capacities for glucose consumption and lactate production evaluated. This treatment strongly decreased the lactate production rate by tumor tissues (85% inhibition) without affecting the other measurements made, i.e. lactate production by control tissues or glucose consumption by both, control and tumor tissues. This result directly correlates with the inhibition promoted by CTZ on the activity of the major regulatory glycolytic enzyme 6-phosphofructo-1-kinase (PFK) that was observed in tumor tissues (84% inhibition) but not in control tissues. Fractionation of the tissues revealed that this inhibition does not occur in the soluble fraction of the enzyme, but is exclusive of a particulate fraction. It has been previously shown that the particulate fraction of PFK activity in tumors is associated to actin filaments (f-actin). Thus, we investigated whether CTZ would affect the association between PFK and f-actin and we found that the drug directly induces the dissociation of the two proteins in the same extent that it inhibits lactate production, total PFK activity and the particulate PFK activity. We concluded that CTZ disrupts glycolysis on human breast tumor tissues, inhibiting PFK activity by dissociating the enzyme from f-actin.

Glioblastoma entities express subtle differences in molecular composition and response to treatment.

Glioblastoma (GBM) is a grade IV astrocytoma. GBM patients show resistance to chemotherapy such as temozolomide (TMZ), the gold standard treatment. In order to simulate the molecular mechanisms behind the different chemotherapeutic responses in GBM patients we compared the cellular heterogeneity and chemotherapeutic resistance mechanisms in different GBM cell lines. We isolated and characterized a human GBM cell line obtained from a GBM patient, named GBM11. We studied the GBM11 behaviour when treated with Tamoxifen (TMX) that, among other functions, is a protein kinase C (PKC) inhibitor, alone and in combination with TMZ in comparison with the responses of U87 and U118 human GBM cell lines. We evaluated the cell death, cell cycle arrest and cell proliferation, mainly through PKC expression, by flow cytometry and western blot analysis and, ultimately, cell migration capability and f-actin filament disorganization by fluorescence microscopy. We demonstrated that the constitutive activation of p-PKC seems to be one of the main metabolic implicated on GBM malignancy. Despite of its higher resistance, possibly due to the overexpression of P-glycoprotein and stem-like cell markers, GBM11 cells presented a subtle different chemotherapeutic response compared to U87 and U118 cells. The GBM11, U87, U118 cell lines show subtle molecular differences, which clearly indicate the characterization of GBM heterogeneity, one of the main reasons for tumor resistance. The adding of cellular heterogeneity in molecular behaviour constitutes a step closer in the understanding of resistant molecular mechanisms in GBM, and can circumvents the eventual impaired therapy.

Value of the intraoperative cytology examination of sentinel lymph node in breast cancer / Valor do exame citológico intraoperatório do linfonodo sentinela no câncer de mama

Objective: Evaluate the value of imprint cytology in the intraoperative analysis of sentinel lymph node (SLN) in patients with breast cancer. Methods: An agreement study for the evaluation of the imprint cytology technique as a diagnostic test for intraoperative SLN among patients diagnosed with breast cancer from January 2007 to January 2017. Results: We studied 210 cases of breast cancer patients submitted to intraoperative sentinel node imprint cytology, aged between 24 and 86 years (mean age 59 years and median age 60 years). The sensitivity of the intraoperative study was 58.3% (95%CI 46.1­69.8%) and the specificity was 97.8% (95%CI 93.8­99.5). The positive predictive value (PPV) was 93.3% (95%CI 81.7­98.6) and the negative predictive value (NPV) was 81.8% (95%CI 75.1­87.4). From the analyzed variables, the presence of macrometastasis was the only one that significantly increased the sensitivity of the imprint to 73.2% (95%CI 59.7­84.2), while micrometastasis presented a sensitivity of only 6.3% (95%CI 0.2­30.2). Conclusion: The use of imprint cytology in the intraoperative SLN study showed good accuracy in predicting axillary status. However, the surgeon and pathologist are fully aware of the set of clinical and histological variables that can influence the sensitivity of the method.

Objetivo: Avaliar o valor do imprint citológico na análise intraoperatória do linfonodo sentinela (LS) em pacientes com câncer de mama. Métodos: Estudo de concordância para avaliação da técnica do imprint citológico como teste diagnóstico do LS no intraoperatório, entre pacientes com diagnóstico de câncer de mama, no período de janeiro de 2007 a janeiro de 2017. Resultados: Foram estudados 210 casos de pacientes com câncer de mama submetidas à citologia de impressão (IC) do linfonodo sentinela no intraoperatório, com idade entre 24 e 86 anos (média de 59 anos e mediana de 60 anos). A sensibilidade do estudo intraoperatório foi de 58,3% (IC95% 46,1­69,8) e a especificidade de 97,8% (IC95% 93,8­99,5). O valor preditivo positivo (VPP) foi de 93,3% (IC95% 81,7­98,6) e o valor preditivo negativo (VPN) de 81,8% (IC95% 75,1­87,4%). Das variáveis analisadas, a presença de macrometástase foi a única que aumentou significativamente a sensibilidade do imprint para 73,2% (IC95%, 59,7­84,2%), enquanto na micrometástase apresentou sensibilidade de apenas 6,3% (IC95% 0,2­30,2). Conclusão: A utilização do imprint citológico no estudo intraoperatório do LS apresentou boa acurácia na previsão do status axilar. Entretanto, é importante o pleno conhecimento, pelo cirurgião e patologista, do conjunto de variáveis clínicas e histológicas que podem influenciar a sensibilidade do método

Sentinel lymph node identification in breast cancer: a comparison study of deep versus superficial injection of radiopharmaceutical.

OBJECTIVES: Sentinel lymph node biopsy (SLNB) has been performed for purposes of disease staging. SLN is usually located in the axillary region and internal mammary chain (IMC). Metastasis in internal mammary nodes can be an important prognostic factor and an indication for systemic treatment in patients with small carcinomas. The SLNB technique continues to evolve and the proper radiopharmaceutical injection route remains under discussion. This study evaluated the success rate of deep injection to identify axillary and extra-axillary SLNs and compared the results with superficial injection technique. METHODS: Forty-six patients diagnosed with breast cancer (stages I and II) were submitted to radiopharmaceutical injection. Deep injection of technetium-99m-dextran 500 was carried out in 20 patients (group A) and periareolar injection of technetium-99m-phytate was carried out in 26 patients (group B). All SLNs were studied by imprint cytology and hematoxylin and eosin staining. RESULTS: SLN identification rate was 76.1% (35 of 46). The SLN identification rate was 75% (15 of 20) for group A and 76.9% (20 of 26) for group B. Axillary SLNs were identified in 65% (13 of 20) of group A and 76.9% (20 of 26) of group B, with no statistical difference (P = 0.75). Extra-axillary SLNs were only identified in group A, and IMC was the principal extra-axillary location. CONCLUSION: Deep injection of radiopharmaceutical achieved a good SLN identification rate in axillary and extra-axillary locations and it is an important method for detecting IMC sentinel nodes.

Asthma and Churg-Strauss syndrome.

We report the case of a 25-year-old woman with Churg-Strauss syndrome, the symptoms of which had first appeared soon after she began taking oral contraceptive at the age of sixteen. The clinical profile evolved rapidly to severe persistent asthma, nasal polyposis, perennial obstructive rhinitis, eosinophilia (peripheral/tissue) and mononeuritis. Churg-Strauss syndrome is the type of disease that demands early detection, accurate diagnosis, aggressive treatment and periodic monitoring. It should be considered in the differential diagnosis of moderate and severe persistent asthma. The case reported calls attention to possibility that there is a hormonal component and that the disease can present early onset.

Asma e síndrome de Churg-Strauss / Asthma and Churg-Strauss syndrome

Relata-se o caso de uma mulher de 25 anos com síndrome de Churg-Strauss, cujos sintomas surgiram aos dezesseis anos, logo após o início do uso de contraceptivo oral. O quadro clínico evoluiu rapidamente com asma persistente grave, polipose nasal, rinite perene obstrutiva, eosinofilia periférica e tecidual, e mononeurite. A síndrome de Churg-Strauss é uma doença que exige suspeita precoce, diagnóstico preciso, tratamento agressivo e monitoramento periódico, devendo ser considerada no diagnóstico diferencial de asma persistente moderada e grave. O caso relatado chama a atenção para possível participação hormonal e surgimento em idade precoce.

We report the case of a 25-year-old woman with Churg-Strauss syndrome, the symptoms of which had first appeared soon after she began taking oral contraceptive at the age of sixteen. The clinical profile evolved rapidly to severe persistent asthma, nasal polyposis, perennial obstructive rhinitis, eosinophilia (peripheral/tissue) and mononeuritis. Churg-Strauss syndrome is the type of disease that demands early detection, accurate diagnosis, aggressive treatment and periodic monitoring. It should be considered in the differential diagnosis of moderate and severe persistent asthma. The case reported calls attention to possibility that there is a hormonal component and that the disease can present early onset.

Valor do exame citológico per - operatório em neurocirurgias / The value of per - operative cytological exam in neurosurgeries

INTRODUÇAO: Mais de 90 por cento dos resultados citológicos per-operatórios neurocirúrgicos correlacionam-se com o histopatológico. Porém, a sua real necessidade é por vezes questionada. OBJETIVO: Avaliar a necessidade do exame per-operatório tendo-se em vista a conduta neurocirúrgica imediata. MATERIAL E MÉTODOS: Revimos 130 casos de exames per-operatórios solicitados de um total de 293 procedimentos cirúrgicos para tratamento de lesões expansivas do sistema nervoso central (SNC), extra ou intra-axiais, sendo 46 biópsias estereotáxicas e 84 cirurgias abertas, realizados de 1998 a 2001. Foram realizados esfregaços, o diagnóstico foi comparado com o histopatológico, e os neurocirurgiões, questionados sobre a implicação do resultado do exame na conduta, caso a caso. RESULTADOS: O exame per-operatório foi solicitado em 44,4 por cento do total de cirurgias, tendo sido considerado relevante para a conduta em 84 por cento destes casos ou, nas biópsias, para avaliar a representatividade do material, o que corresponde a 37,2 por cento do total de cirurgias realizadas no período. Excluindo-se as biópsias estereotáxicas, com indicação inquestionável, o exame foi necessário em 75 por cento, sendo considerado desnecessário, sobretudo nas lesões extra-axiais, em geral solicitado por curiosidade do cirurgião. CONCLUSAO: Apesar de ser um exame com acurácia demonstrada, a realização de exames per-operatórios sem utilidade para a conduta neurocirúrgica pode vir a sobrecarregar patologistas em tarefas desnecessárias. Nossos resultados demonstraram que o exame foi necessário na grande maioria dos casos, mesmo após a exclusão das biópsias estereotáxicas, sobretudo nas lesões intra-axiais, auxiliando na avaliação da demanda desses exames, por vezes excessiva, apesar de útil para treinamento e ensino.

Granulomas hialinizantes pulmonares: aspectos na tomografia computadorizada: relato de caso / Computed tomography findings in patients with pulmonary hyalinizing granulomas: a case report

Granulomas hialinizantes são lesões fibrosantes benignas que em geral se apresentam radiologicamente sob a forma de nódulos múltiplos, freqüentemente escavados e/ou calcificados. Neste trabalho é relatado o caso de uma paciente de 28 anos de idade que apresentou a doença provavelmente secundária a infecção tuberculosa prévia. Ressalta-se a necessidade da inclusão deste diagnóstico diferencial frente ao quadro radiológico de múltiplas lesões nodulares.

Hyalinizing granulomas are benign fibrotic lesions that generally present multiple nodules seen on radiological examinations, which are frequently cavitary and/or calcified lesions. We report a case of a 28 year-old-woman with hyalinizing granulomas probably secondary to a previous tuberculosis infection. Hyalinizing granulomas should be included in the differential diagnosis of patients with multiple pulmonary nodules.

Mastopatia diabética / Diabetic mastopaty

A mastopatia diabética é uma doença rara fibroinflamatória da mama, decorrente da reação auto-imune de acúmulo anormal de proteínas de matriz, causada pela hiperglicemia. Ocorre em pacientes diabéticas de longa data, principalmente as do tipo 1 e que se encontram na pré-menopausa. Aqui se relata um caso de mastopatia diabética em uma mulher de 71 anos de idade, com história de diabetes melito tipo 2, insulino-dependente, de 20 anos de duração, que tinha sido submetida a uma cirurgia plástica mamária aos 35 anos de idade. O exame físico mostrava uma massa endurecida, pouco definida e móvel no quadrante inferior externo, medindo 2 cm. Não possuía expressão mamográfica ou ultra-sonográfica. A biópsia excisional do nódulo e subseqüente exame histopatológico revelaram lobulite linfocítica esclerosante ou mastopatia diabética. O exame clínico pode mimetizar neoplasia maligna de mama e o diagnóstico histopatológico deve ser obrigatório para diagnóstico diferencial.

Diabetic mastopathy is a rare fibroinflammatory breast disease decurrent of autoimmune reaction to the accumulation of abnormal matrix proteins, caused by hyperglycemia. It occurs in long-standing insulin dependent, mainly with type I diabetes, and pre-menopause patients. We present a case of diabetic mastopathy in a 71-year-old woman with a history of insulin-dependent diabetes mellitus type 11 of 20 years duration who has done a mammary plastic surgery. Physical examination showed a stony-hard, ill-defined but freely movable mass at lower outer quadrant measuring 2 cm. It didn't have mammographic netheir ultrassonographic findings. Excisional biopsy of the nodule was performed and the histopathological examination of the lesion revealed sclerosing lymphocitic lobulitis or diabetic mastopathy. Clinical and ultrassonographic exams mimic breast cancer and for this reason the histological diagnosis must be useful for deferential diagnosis.

Adenoma de lactação gigante em gestante adolescente / Giant lactating adenoma in pregnant adolescent

O adenoma da lactação (AL) consiste no tumor sólido mamário mais freqüente em gestantes. Geralmente se apresenta como nódulo móvel, bem delimitado e menor que 3cm, raramente como tumor mamário gigante. Os autores relatam caso de AL em gestante de 15 anos que apresentou rápido desenvolvimento durante o segundo trimestre, tornando necessário tratamento cirúrgico. Apesar do grande volume tumoral, foi possível a realização de incisão periareolar, com resultado estético bastante satisfatório. Esses tumores comumente exibem crescimento durante a gravidez e regressão espontânea após o parto ou a lactação. A hipótese de doença maligna deve sempre ser considerada. Além do carcinoma de mama, o diagnóstico diferencial se faz com o fibroadenoma, o adenoma tubular, o abscesso e a galactocele. A ultra-sonografia é exame bastante útil na avaliação de nódulos mamários no ciclo gravídico-puerperal. A obtenção de amostra para estudo citológico pode ser feita através da punção aspirativa por agulha fina (PAAF). Além desses, a core biópsia constitui método seguro para obtenção de fragmentos para estudo histológico. A biópsia cirúrgica deve ser reservada aos tumores persistentes e volumosos, ou para quando não for possível afastar a hipótese de carcinoma mamário