Methodological aspects of prognostic factor studies: some caveats

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Low educational level but not low income impairs the achievement of cytogenetic remission in chronic myeloid leukemia patients treated with imatinib in Brazil

OBJECTIVES: In Brazil, imatinib mesylate is supplied as the first-line therapy for chronic myeloid leukemia in the chronic phase through the public universal healthcare program, Sistema Único de Saúde (SUS). We studied the socio-demographic factors that influenced therapy success in a population in the northeast region of Brazil. METHODS: Patients with chronic myeloid leukemia from the state of Piauí were treated in only one reference center. Diagnosis was based on WHO 2008 criteria. Risk was assessed by Sokal, Hasford and EUTOS scores. Patients received 400 mg imatinib daily. We studied the influence of the following factors on the achievement of complete cytogenetic response within one year of treatment: age, clinical risk category, time interval between diagnosis and the start of imatinib treatment, geographic distance from the patient's home to the hospital, years of formal education and monthly income. RESULTS: Among 103 patients studied, the median age was 42 years; 65% of the patients had 2-9 years of formal education, and the median monthly income was approximately 100 US$. Imatinib was started in the first year after diagnosis (early chronic phase) in 69 patients. After 12 months of treatment, 68 patients had a complete cytogenetic response. The Hasford score, delay to start imatinib and years of formal education influenced the attainment of a complete cytogenetic response, whereas income and the distance from the home to the healthcare facility did not. CONCLUSION: Patients require additional healthcare information to better understand the importance of long-term oral anticancer treatment and to improve their compliance with the treatment

Computerized texture analysis of atypical immature myeloid precursors in patients with myelodysplastic syndromes: an entity between blasts and promyelocytes

<p>Abstract</p> <p>Background</p> <p>Bone marrow (BM) blast count is an essential parameter for classification and prognosis of myelodysplastic syndromes (MDS). However, a high degree of cell atypias in bone marrow hemopoietic cells may be found in this group of clonal disorders, making it difficult to quantify precisely myeloblasts, and to distinguish them from promyelocytes and atypical immature myeloid precursors. Our aim was to investigate whether computerized image analysis of routine cytology would help to characterize these cells.</p> <p>Methods</p> <p>In May-Grünwald-Giemsa stained BM smears of 30 newly diagnosed MDS patients and 19 cases of normal BM, nuclei of blasts and promyelocytes were digitalized and interactively segmented. The morphological classification of the cells was done by consensus of two observers. Immature granulocytic precursors, which could not be clearly classified either as blasts or promyelocytes, were called "atypic myeloid precursors". Nuclear morphometry and texture features derived from the co-occurrence matrix and fractal dimension (FD) were calculated.</p> <p>Results</p> <p>In normal BM, when compared to myeloblasts, nuclei of promyelocytes showed significant increase in perimeter and local texture homogeneity and a decrease in form factor, chromatin gray levels, Haralick's entropy, inertia, energy, contrast, diagonal moment, cluster prominence, the fractal dimension according to Minkowski and its goodness-of-fit. Compared to normal myeloblast nuclei, the chromatin texture of MDS myeloblasts revealed higher local homogeneity and goodness-of-fit of the FD, but lower values of entropy, contrast, diagonal moment, and fractal dimension. The same differences were found between nuclei of normal promyelocytes and those of MDS. Nuclei of atypical myeloid precursors showed intermediate characteristics between those of blasts and promyelocytes according to the quantitative features (perimeter, form factor, gray level and its standard deviation), but were similar to promyelocytes according to the texture variables inertia, energy, contrast, diagonal moment, cluster prominence, and Minkowski's fractal dimension.</p> <p>Conclusion</p> <p>BM atypical immature myeloid precursors are difficult to be correctly classified in routine cytology. Although their cytoplasm is more similar to that of myeloblasts, computerized texture analysis indicates a nuclear chromatin remodeling more close to the promyelocyte, thus indicating an asynchronous intermediate maturation stage between blast and promyelocyte.</p

Omeprazol e misoprostol na prevenção de lesões de mucosa gástrica causadas por indometacina e celecoxib em ratos

PURPOSE: To evaluate and to compare macro and microscopically the intense injuries of the gastric mucosa of rats which were caused by NSAIDS celecoxib and indomethacin and the gastric cytoprotection with omeprazole and misoprostol. METHODS: The sample is formed by one hundred and fifty Wistar rats with average weight 200 g, distributed in four groups, such as: Group A, subdivided in groups A1 and A2 - pre-treatment with omeprazole (20 mg/rat) during seven days and on the 8th day - use of NSAIDS, concerning A1 (20 rats) were given celecoxib (1mg/rat) and A2 (20 rats) were given indomethacin. The Group B, subdivided in group B1 and B2 - pre-treatment with misoprostol (20mg/rat) during seven days and on the 8th day use of NSAIDS, concerning B1 (20 rats) were given celecoxib (1 mg/ rat) and B2 (20 rats) were given indomethacin (12.5 mg/rat). The Group C: were not given cytoprotection during seven days, from the 7th to the 8th day - fast of food and water ad libitum, on the 8th day of NSAIDS use, concerning C1 (20 rats) were given celecoxib, C2 (20 rats) were given indomethacin (12.5 mg/ rat), C3 (20 rats) were given celecoxib (200mg/rato), and Group D - control group, concerning 10 rats were observed during seven days ingesting food and water ad libitum. On the 9th day, the stomachs were taken out and were macro and microscopically evaluated for the identification of the gastric injuries. RESULTS: On the macroscopic studies, the groups A2, B2 and C2 presented a remarkable high number of injuries for cm² /animal, respectively 18.55 injuries for cm² /animal, 16.25 injuries for cm² /animal and 13.55 injuries for cm²/animal. On the microscopic studies, the percentage of the injured mucosa, presented expressive difference among the groups A1, B1, C1 when compared to the groups A2, B2, C2 (p<0.0001). The average of the length/injury and the average of the depth of the injuries did not present expressive statistics differences among the groups A2, B2 and C2. The average of the edema presented expressive statistics difference among the groups A2 and D; B2 and C2 and between C2 and D (p < 0.05). CONCLUSIONS: The indomethacin on the applied concentration causes a great number of macroscopic and microscopic injuries to gastric mucosa of rats when compared to celecoxib which does not cause lesions. Omeprazole and misoprostol on the applied concentrations do not present macroscopic and microscopic effectiveness on the gastric cytoprotection when applying indomethacin. Considering the microscopic analysis of the average of the edema, the group of animals, which was given misoprostol as cytoprotection, presented a lower average compared to the group which was given omeprazole.OBJETIVO: Avaliar e comparar macro e microscopicamente as lesões agudas da mucosa gástrica de ratos provocadas pelos AINEs celecoxib e indometacina e a citoproteção gástrica com omeprazol e misoprostol. MÉTODOS: A amostragem consistiu 150 ratos machos da raça Wistar, com peso médio de 200g, divididos em quatro grupos, a saber: grupo A, subdividido em grupos A1 e A2 - pré-tratamento com omeprazol (20 mg/rato) durante sete dias, e no oitavo dia receberam o AINEs, sendo A1 (20 ratos) receberam celecoxib (1mg/rato) e A2 (20 ratos) receberam indometacina (12,5mg/rato). O grupo B, subdividido em grupo B1 e B2 - pré-tratamento com misoprostol (20ìg/rato) durante sete dias e no oitavo dia receberam AINEs, sendo B1 (20 ratos) receberam celecoxib (1mg/rato) e B2 (20 ratos) receberam indometacina (12,5mg/rato). O grupo C não recebeu citoproteção durante sete dias e no oitavo dia recebeu AINEs, sendo C1 (20 ratos) receberam celecoxib (1mg/rato) , C2 (20ratos) receberam indometacina (12,5mg/rato), C3 (20 ratos) receberam celecoxib e grupo D - grupo controle, no qual dez ratos foram observados recebendo ração e água ad libitum. A seguir, no 9º dia (de todos os grupos), os estômagos eram removidos e avaliados macro e microscopicamente para a identificação das lesões gástricas. RESULTADOS: Na análise macroscópica, os grupos A2, B2 e C2 apresentaram número de lesões por cm²/animal significativamente elevados, sendo respectivamente 18,55 lesões por cm²/animal, 16,25 lesões por cm²/animal e 13,55 lesões por cm²/animal. Na análise microscópica, a porcentagem da mucosa com lesão mostrou diferença significativa entre os grupos A1, B1, C1 quando comparados com os grupos A2, B2 e C2 (p<0,0001). Os resultados da média da extensão/lesão e da média da profundidade das lesões não mostraram diferenças estatísticas significativas entre os grupos A2, B2 e C2. A média do edema mostrou diferença estatística significativa entre os grupos A2 e D; B2 e C2 e entre C2 e D (p<0,05). CONCLUSÕES: A indometacina na concentração empregada provoca número significativo de lesões macro e microscópicas na mucosa gástrica de ratos quando comparadas ao celecoxib, que não provocou lesões. O omeprazol e o misoprostol nas concentrações empregadas não mostraram efetividade macro e microscópica na citoproteção gástrica à administração da indometacina. Considerando-se a análise microscópica da média do edema, o grupo de animais que recebeu misoprostol como citoprotetor apresentou menor média em comparação ao grupo que recebeu o omeprazol.16817

D2-40, um novo marcador endotelial linfático: identificação de invasão linfática e relação com metástases axilares em câncer de mama

BACKGROUND: Studies of lymphatic vessels were limited by the lack of specific markers. Recently, they have become possible due to the release of new D2-40 antibody, a selective marker for lymphatic endothelium. The aim of our study was to compare neoplastic invasion in lymphatic and blood vessels detected in hematoxylin and eosin (H&E) and immunohistochemistry-stained sections. METHODOLOGY: A total of 123 cases of invasive mammary carcinomas were studied and sorted out into three subgroups according to axillary staging (macrometastasis, micrometastasis and lymph node negative). Lymphatic vessel invasion (LVI) and blood vessel invasion (BVI) were initially evaluated in histological H&E and immunohistochemistry-stained sequential sections. Lymphatic and blood vessel invasions were assessed by immunohistochemistry, employing D2-40 and CD31 antibodies, respectively. LVI and BVI were related to size, type, histologic grade of primary tumors, and the presence of metastasis. RESULTS: LVI was detected through H&E staining procedure in 17/123 cases (13.8%), and through immunohistochemistry procedure in 35/123 cases (28.5%) (kappa = 0.433). BVI was detected through H&E in 5/123 cases (4.1%), and through immunohistochemistry in 19/123 cases (15.4%) (kappa = 0.198). LVI and BVI were positively related to higher histologic grade of primary tumors (p < 0.05). LVI was also positively related to the presence of macrometastasis. CONCLUSION: The detection of lymphatic and blood vessel invasions through immunohistochemistry employing D2-40 and CD31 was higher than the detection through H&E, and it was related to higher tumor grade and metastasis in axillary lymph nodes.INTRODUÇÃO: Estudos de vasos linfáticos eram limitados pela ausência de marcadores endoteliais linfáticos específicos. Recentemente, eles se tornaram possíveis após liberação comercial do novo anticorpo D2-40, marcador seletivo para endotélio linfático. O objetivo do nosso estudo foi comparar invasão neoplásica em vasos linfáticos e sanguíneos detectada em secções coradas pela hematoxilina e eosina (HE) e imuno-histoquímica (IIQ). MATERIAIS E MÉTODOS: Foram estudados 123 casos de carcinomas mamários invasores subdivididos em três subgrupos de acordo com o estadiamento axilar: macrometástases (Mac-Met), micrometástases (Mic-Met) e linfonodo negativo (LNN). Invasão de vasos linfáticos (IVL) e de vasos sangüíneos (IVS) foi inicialmente avaliada em secções histológicas coradas pela HE e através da IIQ realizada em cortes seqüenciais. A invasão de vasos linfáticos e sanguíneos foi avaliada pela imuno-histoquímica, empregando-se respectivamente os anticorpos D2-40, e CD31. IVL e IVS foram relacionadas com tamanho tumoral, tipo e grau histológico dos tumores primários e com a presença de metástases. RESULTADOS: IVL foi observada pela HE em 17/123 casos (13,8%) e pela IIQ em 35/123 casos (28,5%) (kappa = 0,433). IVS foi observada pela HE em 5/123 casos (4,1%) e pela IIQ em 19/123 casos (15,4%) (kappa = 0,198). IVL e IVS estavam positivamente relacionadas com maior grau histológico dos tumores primários (p < 0,05). IVL também estava positivamente relacionada com a presença de macrometástases. CONCLUSÃO: A detecção IIQ, respectivamente por D2-40 e CD31, de invasão de vasos linfáticos e sanguíneos foi maior que a detecção feita em cortes corados pela HE e relacionou-se com maior grau tumoral e metástases em linfonodos axilares.455

Photodynamic Effect Produced By Hene Radiation In Harderian Glands Of Wistar Rats: An Experimental Model For Pdt Studies

In rats, the Harderian Gland secret Protoporphirin IX which is retained at acinar lumina. Since this photosensitizer is important for PDT of malignant tumors, we propose to study this gland as a model to help understanding PDT with endogenous photosensitizers. Twenty Wistar SPF adult rats were submitted to surgical exposure of both Harderian glands, revealing red fluorescence upon UV, characterizing the protoporphirin IX presence. After that, one gland of each pair (one kept as control) was irradiated with an 8mW HeNe (6328 angstron) for 45 minutes, delivering about 2.7 joules/mm 2. After 24 hours a group of 10 animals were sacrificed and the glands removed for histological analysis. The remaining animals were subjected to the same procedure but the glands were removed immediately after laser treatment. Histological and fluorescence analysis immediately after laser irradiation showed cell fragmentation with loss of acinar architecture with diffusion of protoporphirin in the cytoplasm of damaged cells, as well as interstitial edema. After 24 hours these alterations were more pronounced with accentuated loss of intraluminal protoporphirin and beginning of leukocytic demarcation of necrotic areas. The innate Harderian glands of rats, exposed to HeNe laser, showed a similar behavior as tumor tissue under PDT.3914404

Angiogenesis in the progression of cutaneous squamous cell carcinoma: an immunohistochemical study of endothelial markers

OBJECTIVE: To demonstrate the role of angiogenesis in the progression of cutaneous squamous cell carcinoma. INTRODUCTION: Angiogenesis is a pivotal phenomenon in carcinogenesis. Its time course in cutaneous squamous cell carcinoma has not yet been fully established. METHODS: We studied the vascular bed in 29 solar keratoses, 30 superficially invasive squamous cell carcinomas and 30 invasive squamous cell carcinomas. The Chalkley method was used to quantify the microvascular area by comparing panendothelial (CD34) with neoangiogenesis (CD105) immunohistochemical markers. The vascular bed from non-neoplastic adjacent skin was evaluated in 8 solar keratoses, 10 superficially invasive squamous cell carcinomas and 10 invasive squamous cell carcinomas. RESULTS: The microvascular area in CD105-stained specimens significantly increased in parallel with cutaneous squamous cell carcinoma progression. However, no differences between groups were found in CD34 sections. Solar keratosis, superficially invasive squamous cell carcinoma and invasive squamous cell carcinoma samples showed significant increases in microvascular area for both CD34- and CD105-stained specimens compared with the respective adjacent skin. DISCUSSION: The angiogenic switch occurs early in the development of cutaneous squamous cell carcinoma, and the rate of neovascularization is parallel to tumor progression. In contrast to panendothelial markers, CD105 use allows a dynamic evaluation of tumor angiogenesis. CONCLUSION: This study demonstrated the dependence of skin carcinogenesis on angiogenesis