Leucemia linfoide aguda a importancia do laboratorio de liquor para o sucesso do tratamento / Acute lymphoblastic leukemia: the importance of cerebrospinal fluid laboratory for successful treatament

A Leucemia Linfoide Aguda (LLA) e uma doenca caracterizada pelo acumulo de linfoblastos em numerosos orgaos e tecidos, notadamente na medula ossea. Entretanto as celulas malignas da LLA tem uma predisposicao de infiltrar o Sistema Nervoso Central (SNC) e os testiculos, sendo estes, considerados ¡°santuarios¡±. A importancia ao diagnostico da avaliacao citologica do liquido cefalorraqueano(Liquor) tornou-se fundamental para adequacao do tratamento, prognostico e para o monitoramento de eventuais recaidas. Citologicamente pode-se determinar um ¡°STATUS¡±, sendo que a avaliacao mais aceita atualmente ao diagnostico deve seguir os seguintes criterios: Status 1: puncao nao traumatica com ausencia de blastos apos citocentrifugacao. Status 2: puncao nao traumatica com presenca de blastos apos citocentrifugacao e leucocitos ¡Ü5/mm3. Status 3: puncao nao traumatica com presenca de blastosapos citocentrifugacao e leucocitos ¡Ý5/mm3 . A puncao traumatica deve ser classificada como risco, pois pode haver a infiltracao na hora da puncao. O objetivo deste trabalho e definir criteriosamente a importancia da atuacao do Farmaceutico Bioquimico no Laboratorio de Liquor auxiliando o clinico na avaliacao de conduta terapeutica baseado na avaliacao citologica do liquido cefalorraqueano.

Acute Lymphoblastic Leukemia (ALL) is an illness characterized for the accumulation of blasts in numerous organ and tissue, essential in the blone marrow. However the malignant cells of the ALL have a predisposition to infiltrate central nervous system(CNS) and the testicules, being been these, considered "sanctuaries". The importance to the diagnosis of the cytological evaluation of the cerebrospinal fluid (CSF), became basic for adequacy of the treatment, prognostic and for the involvement of eventual fallen again. Cytologically a "STATUS" can be determined, being most accepted currently to the diagnosis must follow the following criteria:CNS1 (puncture nontraumatic without leukemic blasts after ytocentrifugation), CNS2 (puncture nontraumatic, ¡Ü5 WBC/mm3 CSF with identifiable blasts after cytocentrifugation), CNS3 (puncture nontraumatic, ¡Ý5 WBC/mm3 CSF with identifiable blasts aftercytocentrifugation). TLP(+) ¨C puncture traumatic with blasts, and TLP(-) ¨C puncture traumatic without blasts. The traumatic puncture must be classified as risk therefore it can have infiltration in the hour puncture. The objective of this work is reintensification to define the importance of the performance of the Pharmaceutical Biochemist in the Cerebrospinal Fluid Laboratory assisting the physician in the based evaluation of therapeutical behavior in the cytological evaluation of the Cerebrospinal Fluid.

Quantification of cerebrospinal fluid ferritin as a biomarker for CNS malignant infiltration / Quantificação de ferritina no líquido cefalorraquidiano como biomarcador para o diagnóstico de infiltração maligna no SNC

Several markers have been studied for their ability to make the CNS infiltration diagnosis earlier and more precise; previous studies showed that CSF ferritin concentrations were higher in patients with malignant invasion of CNS. The objective was to determine the importance of CSF ferritin as a biomarker for the diagnosis of CNS neoplasic infiltration. This study is based on 93 CSF samples, divided into five groups: malignant cells present (n13); malignant cells not present (n26); inflammatory neurological diseases (n16); neurocysticercosis (n20); acute bacterial meningitis (n18). CSF ferritin values were determined by micro particle enzyme immunoassay. CSF ferritin level (mean±SD) in the group with neoplasic cells in the CSF was 42.8±49.7 ng /mL, higher than in the other groups (p<0.0001). We conclude that CSF ferritin with the cut off 20 ng/mL could be an adjuvant biomarker to the diagnosis of CNS malignant infiltration.

Diversos marcadores foram estudados com a finalidade de avaliar sua capacidade de diagnosticar a infiltração neoplásica no SNC precocemente e de forma mais precisa. Estudos anteriores mostraram que as concentrações de ferritina no LCR eram mais elevadas nos pacientes com infiltração neoplásica no SNC. O objetivo foi determinar a importância da ferritina no LCR como biomarcador para o diagnóstico de infiltração neoplásica no SNC. Este estudo é baseado em 93 amostras do LCR, divididas em cinco grupos: células malignas presentes (n13); células malignas ausentes (n26); doenças neurologicas inflamatórias (n16); neurocisticercose (n20); meningites bacterianas agudas (n18). Os valores de ferritina no LCR foram determinados por ELISA de microparticulas. O nível de ferritina no LCR (média±desvio padrão) no grupo com células neoplásicas no LCR foi 42,8±49,7 ng/mL, mais elevado do que nos outros grupos (p<0.0001). Concluímos que a ferritina no LCR com cut off de 20 ng/mL pode ser um biomarcador para o diagnóstico de infiltração maligna no SNC.

Cerebrospinal fluid cytological and biochemical characteristics in the presence of CNS neoplasia

Central nervous system (CNS) infiltration must be ruled out in patients with known neoplastic diseases and neurological symptoms. It was done a retrospective analysis of 1,948 CSF samples from patients with suspected malignant infiltration in the CNS, in order to evaluate the positivity rate of malignant cells in cerebrospinal fluid (CSF) samples and correlate with cytochemical characteristics. Sixty-two percent of subjects had acute lymphocytic leukemia. Malignant cells were found in 24 percent of all CSF samples. Subjects with positive malignant cells had predominance of increased levels of CSF total protein (TP), glucose and total cytology (p<0.05). Mean total cell count in this group was 232 (SD 933) cells/mm³, compared to 9 (SD 93) cells/mm³ in the group without neoplasic cells (p=0.029). CSF TP specificity was 87 percent and negative predictive value (NPV) 96 percent. CSF total cell count specificity 86 percent and NPV 97 percent. Although sensitivity and positive predictive value were low. The presence of inflammatory cells and elevated TP found in patients with malignant cells in the CSF can aid in diagnosing CNS neoplasms.

A infiltração neoplásica no SNC deve ser afastada em pacientes com neoplasia e sintomas neurológicos. Foi realizada uma análise retrospectiva de 1.948 amostras de LCR de pacientes com suspeita de infiltração neoplásica no SNC. Sessenta e dois por cento dos pacientes eram portadores de leucemia linfocitica aguda. Células neoplásicas foram encontradas em 24 por cento de todas as amostras. Houve níveis aumentados no LCR da proteína total (PT), glicose e de citologia global (p<0.05), no grupo com presença de células neoplásicas. A média da contagem global de células no LCR, neste grupo, foi 232±933 cels/mm³, contra 9±93 cells/mm³ no grupo sem células neoplásicas no LCR (p=0,029). O aumento de PT no LCR apresentou especificidade 87 por cento e valor preditivo negativo (VPN) 96 por cento. A contagem global de células no LCR apresentou especificidade 86 por cento e VPN 97 por cento. Porém sensibilidade e valores preditivos positivos foram baixos. A presença de células inflamatórias e PT no LCR elevada em pacientes com neoplasias pode ser um indicador do envolvimento no SNC.

Lavado y cepillado bronquiales en tumores pulmonares centrales y periféricos sospechosos de malignidad / Bronchial washing and brushing in central and peripheral lung masses suspicious of malignancy

El diagnóstico citológico e histopatológico en casos de tumores pulmonares se realiza con alta confiabilidad a través de muestras obtenidas por fibrobroncoscopía (FBC). Puede haber diferencias que dependen de la localización del tumor. Objetivo: Determinar la sensibilidad del lavado, cepillado bronquial y biopsia por (FBC) en una muestra de tumores pulmonares centrales, donde hay alteraciones endoscópicas evidentes y en otra de tumores pulmonares periféricos con sospecha de malignidad donde las alteraciones son menos frecuentes. Casos: Se estudiaron 86 enfermos con tumores pulmonares, 44 centrales (Grupo A) y 42 periféricos (Grupo B). Todos fueron objeto de estudio por FBC con lavado y cepillado; en 15 casos se practicó biopsia. Resultados: Hubo una diferencia significativa (p < 0.005) con respecto a la apariencia normal y anormal entre tumores centrales (A) y periféricos (B). El diagnóstico de malignidad basado en la presencia de células malignas por cepillado y lavado bronquiales fue positivo en 21 del Grupo A y en 14 del Grupo B con sensibilidad de 60 y 40%, respectivamente. En 15 casos de tumor endobronquial se practicó biopsia. En los casos negativos, 23 del A y 28 del B, se emplearon otros métodos diagnósticos (p < 0.05). Se detectaron 76 casos de neoplasias, predominando el carcinoma bronquiogénico (43%). Diez casos fueron procesos infecciosos. Conclusión: El estudio por FBC permanece como un importante método diagnóstico en casos de tumores pulmonares. El lavado y el cepillado tuvieron resultados positivos en 35/86, cifra relativamente baja que sugiere la necesidad de mejorar la calidad de las muestras obtenidas. La negatividad por FBC obliga el empleo de otros métodos diagnósticos. El costo estimado de los procedimientos, erogado por el paciente en dólares americanos, es notablemente menor que en países como Holanda, que se tomó como comparativo.

Cytologic and histologic diagnosis of lung tumors can usually be done by means of fiberoptic bronchoscopy (FOB), but there are some differences in cases of central or peripheral tumors. Objective: To determine the sensititivity of bronchial brushing, lavage and biopsy performed by FBO in a sample of central tumors, with evident bronchial alterations, and another sample of peripheral lesions in which these alterations are less frequent. A preliminary comparison of the costs of FOB in Mexico and Holland was also done. Cases: There were 86 patients with tumoral lesions suspicious of malignancy, 44 central (Group A) and 42 peripheral (Group B); all were subjected to FOB, lavage and bronchial brush ings; biopsy was done in 15 cases of endobronchial lesions. Results: There was a significat difference (p < 0.005) as to the normal or abnormal appearance of the bronchial mucosa between central tumors (A) and peripheral (B) lesions. Diagnosis of malignancy by lavage and brushing based in the finding of malignant cells was positive in 21 of the Group A and in 14 of Group B, sensitivity of 60% and 40% respectively. Biopsy was performed in 15 cases with endobronchial tumor. In the negative cases, 23 in Group A and 28 in Group B, other diagnostic methods were employed (p < 0.05). A total of 76 cases of malignancy were detected; bronchogenic carcinoma was predominant (43%). Ten cases of infectious diseases were identified. Conclusion: FOB remains as an important diagnostic tool in cases of lung tumors. Bronchial lavage and brushing had positive results for malignant cells in 35/86; this relative low figure suggests the need to improve the quality of the samples obtained by FOB. Other diagnostic methods must be used in cases with negative FOB results. Estimated costs, in US dollars, of diagnostic methods are much lower in Mexico than in an European country, The Netherlands.

The Significance of AgNOR Count in Body Fluid: Differential between reactive mesothelial cells & malignant cells / 대한세포병리학회지

To distinguish reactive mesothelial cells from malignant cells in body fluid, we applied silver staining of nucleolar organizer regions(AgNORs) to ethanol fixed cytologic preparations. Fifty aspirated samples of benign(22 cases) and malignant(26 cases) body fluids were studied using the one step silver staining method. Two cytologically atypical samples were also included in the study. In malignant cases the mean AgNOR count was 3.56+/-0.81, while in benign cases the mean AgNOR count was 2.02+/-0.33. The difference of AgNOR counts between these two groups were statistically significant(p<0.001). The mean of atypical cases was 2.91. Both were diagnosed as malignant in follow-up cytology. In malignant effusions, there is statistically significant difference in AgNOR counts between cells forming complex papillae or clusters and singly scattered cells(p<0.05), 3.29+/-0.95 and 3.83+/-0.55, respectively. We concluded that AgNOR count appears to be useful as a diagnostic tool especially when the cytologic differentiation is difficult.

Electron Microscopic Studies on Invasion of Malignant Cells to Smooth Muscle Cells / 昆明医科大学学报

The invasion of malignant cells into smooth muscle cells was shown by EM.We dicsovered that the malignant cells adhered to and penetrated the smooth muscle cells with their microvilli and pseudopodia.The microfilament in the microvilli and pseudopodia is the motion skeleton of cancer cells.Studying the structure and function of the microfilament fur- ther is important for researching the metastasis of tumor.

Electron Microscopic Studies on Invasion of Malignant Cells to Collagen / 昆明医科大学学报

The degradation of collagen in the extracellular matrix induced by malignant cells is a very impotant link in the cancerometastasis chain.The invasive and destructive effect of vari- ous types of malignant cells cultured in vitro on the three dimensional gel of the native collagen fibrils was observed by electron microscopy.The cytobiological properties of malig- nant cells were discussed and compared with the normal ones.

PREPARATION OF MONOCLONAL ANTIBODY AGAINST PROTEIN KINASE A AND ITS IMMUNOCYTOCHEMICAL LOCALIZATION IN NORMAL AND MALIGNANT CELLS / 解剖学报

A rat monoclonal antibody of the IgG1 class, McAb B5A8, specific for the cAMP-dependent protein kinase(PKA) was produced. Western blot analysis revealed specific binding of the antibody to protein of 52-56 kd.The affinity-purified McAb BSA8 was labeled with FITC. Immunofluorescent localization of PKA was examined in human fibroblasts, gastric cancer cell line (MGc 80-3)and EAC cells. The distribution of PKA on the cytoplasmic microtubule network, Golgi region and nucleoli were observed. PKA was localized in the nuclear region in G2 phase of synchronized MGc 80-3 cells, and it was only found around the nuclei of MGc 80-3 cells which were incubated with DBcAMP. The changes in the distribution of PKA in cells are discussed.