RESUMO
BACKGROUND: Chronic lymphocytic leukaemia (CLL) is a neoplasm of B-cells characterized by variable prognosis. Exploring the proteome of CLL cells may provide insights into the disease. Therefore, eleven proteomics experiments were conducted on eleven primary CLL samples. RESULTS: We reported a CLL proteome consisting of 919 proteins (false discovery rate (FDR) 1%) whose identification was based on the sequencing of two or more distinct peptides (FDR of peptide sequencing 1%). Mass spectrometry-based protein identification was validated for four different proteins using Western blotting and specific antibodies in different CLL samples. Small sizes of nucleolin (~57 kDa and ~68 kDa) showed a potential association with good prognosis CLL cells (n = 8, p < 0.01). Compared with normal B-cells, CLL cells over-expressed thyroid hormone receptor-associated protein 3 (THRAP3; n = 9; p = 0.00007), which is implicated in cell proliferation; and heterochromatin protein 1-binding protein 3 (HP1BP3; n = 10; p = 0.0002), which promotes cell survival and tumourogenesis. A smaller form of HP1BP3, which may correspond to HP1BP3 isoform-2, was specifically identified in normal B-cells (n = 10; p = 0.0001). HP1BP3 and THRAP3 predicted poor prognosis of CLL (p 0.05). Consistently, THRAP3 and HP1BP3 were found to be associated with cancer-related pathways (p 0.05). CONCLUSIONS: Our findings add to the known proteome of CLL and confirm the prognostic importance of two novel cancer-associated proteins in this disease.
Assuntos
Leucemia Linfocítica Crônica de Células B , Biomarcadores Tumorais/análise , Espectrometria de Massas , Fatores de Transcrição/análise , Proteínas Nucleares/análise , Western Blotting , Cromatografia Líquida , Proteômica , Proteínas de Ligação a DNA/análiseRESUMO
ABSTRACT BACKGROUND: There are cases of colorectal tumors that, although small, show more aggressive evolution than large tumors. This motivated us to study whether there are any proteins capable of alerting about these changes. The aim here was to correlate the immunoexpression of the TS, p53, COX2, EGFR, MSH6 and MLH1 biomarkers in tumors in patients with colorectal adenocarcinoma, with the degree of cell differentiation, tumor staging and clinical-pathological prognostic factors. DESIGN AND SETTING: Retrospective observational study at a public tertiary-level hospital. METHODS: We analyzed tissue-microarray paraffin blocks of tumor tissues that had been resected from 107 patients. We used Fisher's exact test to study associations between tumor differentiation/staging and the immunoexpression of biomarkers. We also used Kaplan-Meier estimation, the log-rank test and the adjusted Cox regression model to investigate the patients' overall survival (in months) according to biomarkers and disease-free interval. RESULTS: The degree of tumor differentiation and tumor staging were not associated with the biomarkers, except in cases of patients in stages III or IV, in which there was a correlation with MLH1 expression (P=0.021). Patient survival and disease-free interval were not associated with the biomarkers. CONCLUSION: There were no associations between the degree of tumor differentiation, staging, length of survival or disease-free interval and the immunoexpression of the TS, p53, COX2, EGFR or MSH6 tumor markers. In advanced cases of colorectal adenocarcinoma (stages III and IV), there was a higher percentage of MLH1-negative results.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Neoplasias Colorretais/mortalidade , Neoplasias Colorretais/patologia , Adenocarcinoma/patologia , Biomarcadores Tumorais/análise , Valores de Referência , Timidilato Sintase/análise , Imuno-Histoquímica , Adenocarcinoma/mortalidade , Modelos de Riscos Proporcionais , Estudos Retrospectivos , Estudos Longitudinais , Proteína Supressora de Tumor p53/análise , Análise Serial de Tecidos , Proteínas de Ligação a DNA/análise , Ciclo-Oxigenase 2/análise , Estimativa de Kaplan-Meier , Receptores ErbB/análise , Proteína 1 Homóloga a MutL/análise , Estadiamento de NeoplasiasAssuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Carcinoma Pulmonar de Células não Pequenas/química , Proteínas de Ligação a DNA/análise , Endonucleases/análise , Neoplasias Pulmonares/química , Fatores de Tempo , Imuno-Histoquímica , Cisplatino/uso terapêutico , Resultado do Tratamento , Paclitaxel/uso terapêutico , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/tratamento farmacológico , Antineoplásicos/uso terapêuticoRESUMO
ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.
Assuntos
Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Adenocarcinoma/patologia , Células Epiteliais/patologia , Células-Tronco Neoplásicas/patologia , Neoplasias da Próstata/patologia , Células Estromais/patologia , Actinas/análise , Adenocarcinoma/química , Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Progressão da Doença , Proteínas de Ligação a DNA/análise , Células Epiteliais/química , Receptor alfa de Estrogênio/análise , /análise , Proteínas Ligadas por GPI/análise , Imuno-Histoquímica , Fator de Crescimento Insulin-Like I/análise , /análise , Gradação de Tumores , Proteínas de Neoplasias/análise , Células-Tronco Neoplásicas/química , Neoplasias da Próstata/química , Células Estromais/química , Microambiente Tumoral , Fatores de Transcrição/análise , Vimentina/análiseRESUMO
Introductıon Ureteral obstruction is a common pathology and caused kidney fibrosis and dysfunction at late period. In this present, we investigated the antifibrotic and antiinflammatory effects of montelukast which is cysteinyl leukotriene receptor antagonist, on kidney damage after unilateral ureteral obstruction(UUO) in rats. Mateirıals and Methods 32 rats divided four groups. Group 1 was control, group 2 was sham, group 3 was rats with UUO and group 4 was rats with UUO which were given montelukast sodium (oral 10 mg/kg/day). After 14 days, rats were killed and their kidneys were taken and blood analysis was performed. Tubular necrosis, mononuclear cell infiltration and interstitial fibrosis scoring were determined histopathologically in a part of kidneys; nitric oxide(NO), malondialdehyde(MDA) and reduced glutathione(GSH) levels were determined in the other part of kidneys. Urea-creatinine levels were investigated at blood analysis. Statistical analyses were made by the Chi-square test and one-way analysis of variance (ANOVA). Results There was no difference significantly for urea-creatinine levels between groups. Pathologically, there was serious tubular necrosis and fibrosis in group 3 and there was significantly decreasing for tubular necrosis and fibrosis in group 4(p<0.005). Also, there was significantly increasing for NO and MDA levels; decreasing for GSH levels in group 3 compared the other groups(p<0.005). Conclusıon We can say that montelukast prevent kidney damage with antioxidant effect, independently of NO. .
Assuntos
Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias da Mama/química , Carcinoma Ductal de Mama/química , Proteínas de Ligação a DNA/análise , Transição Epitelial-Mesenquimal , Receptor alfa de Estrogênio/análise , Fatores de Transcrição/análise , Biomarcadores Tumorais/análise , Neoplasias da Mama/patologia , Caderinas/análise , Carcinoma Ductal de Mama/patologia , Imuno-Histoquímica , Valor Preditivo dos Testes , Prognóstico , Análise Serial de Tecidos , beta Catenina/análiseRESUMO
This study centers on relationships among national and international actors in preparation of the first health policy document for East Timor, under the United Nations transitional administration, between 1999 and 2002. International cooperation support for the health system rehabilitation process during the post-conflict period is analyzed as part of reconstruction of the State in parallel with construction of the country's political and institutional framework. Knowledge, ideas, "ways of doing," and induced and accepted practices permeate an interplay of power relationships that condition both national political alliance-building and the architecture of international aid, pointing to input to a discussion of how these mechanisms interact at different conjunctures and times in different negotiating frameworks. .
Dedica-se, aqui, às relações entre diferentes atores na elaboração do primeiro documento de política de saúde para o Timor-Leste, sob a administração transitória das Nações Unidas, de 1999 a 2002. O apoio da cooperação internacional no processo de reabilitação do sistema de saúde no período pós-conflito é analisado como parte da reconstrução do Estado e concomitante à construção do arcabouço político e institucional no país. Conhecimentos, ideias, "modos de fazer" e práticas induzidas e aceitas entremeiam um jogo de relações de poder que condiciona tanto a articulação política nacional quanto a arquitetura da ajuda externa, apontando elementos para a discussão de como esses mecanismos se organizam em conjunturas diferentes de negociação.
Assuntos
Humanos , Antígenos de Neoplasias/análise , Carcinoma de Células Escamosas/química , DNA Topoisomerases Tipo II/análise , Proteínas de Ligação a DNA/análise , Neoplasias de Cabeça e Pescoço/química , Imuno-Histoquímica , /análise , Mucosa/química , Lesões Pré-Cancerosas/química , Biomarcadores Tumorais/análise , Biópsia , Estudos de Casos e Controles , Carcinoma de Células Escamosas/patologia , Diagnóstico Diferencial , Progressão da Doença , Neoplasias de Cabeça e Pescoço/patologia , Mucosa/patologia , Valor Preditivo dos Testes , Lesões Pré-Cancerosas/patologia , Fatores de TempoRESUMO
Many predictive models have been proposed for better stratification of diffuse large B-cell lymphoma (DLBCL). Hans' algorithm has been widely used as standard to sub-classify DLBCL into germinal center B-cell (GCB) and non-GCB origins. However, there have been disagreements in the literature regarding its prognostic significance. Here, we retrospectively analyzed Hans' algorithm and the individual immunohistochemical biomarkers at different cut-off values (5%, 30%, 50% or 75%) in 94 Korean patients with DLBCL treated with combination chemotherapy with cyclophosphamide, daunorubicin, vincristine, and prednisone. No significant differences were observed between the GCB (18 patients, 19.1%) and non-GCB (76, 80.9%) groups. Among individual biomarkers, CD10 negativity (cut point: 30%) and bcl-6 positivity (cut point: 5%) were independent good prognostic markers in progression-free survival (PFS), whereas bcl-6 (cut point: 5%) positivity was an independent good prognostic marker in overall survival irrelevant of international prognostic index. The present study showed the lack of predictability of Hans' algorithm in DLBCL patients, and that CD10, Bcl-6 may have diverse prognostic significance at different cut-off values. Our results suggest that the proposed cut-off value may not be applied universally, and that the optimal cut-off value may need to be optimized for individual laboratory.
Assuntos
Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Algoritmos , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Ciclofosfamida/uso terapêutico , Proteínas de Ligação a DNA/análise , Doxorrubicina/uso terapêutico , Linfoma Difuso de Grandes Células B/classificação , Neprilisina/análise , Prednisona/uso terapêutico , Prognóstico , República da Coreia , Estudos Retrospectivos , Biomarcadores Tumorais/análise , Vincristina/uso terapêuticoRESUMO
Apoptose, proliferação e histomorfometria do baço foram investigados em ratas Wistar adultas ovariectomizadas e não-ovariectomizadas, mantidas em hipotireoidismo induzido pela administração diária de propiltiouracil (PTU) por 120 dias. Dois grupos eutireóideos ovariectomizados e não-ovariectomizados serviram como controle. Foi colhido o plasma para dosagem de T4 livre e o baço para análise da histomorfometria, do índice apoptótico e da expressão imunohistoquímica de caspase 3 e CDC47. Valores de T4 livre foram menores nas ratas tratadas com PTU (p < 0,05). Nos grupos hipotireóideos houve redução do peso do baço, do número e do tamanho dos folículos linfóides e aumento do índice apoptótico e da expressão de caspase 3 (p < 0,05). Porém, o baço de ratas hipotireóideas ovariectomizadas apresentou aumento menos acentuado do índice apoptótico e da expressão de caspase 3 do que o baço de ratas hipotireóideas não-ovariectomizadas (p < 0,05). O grupo eutireóideo ovariectomizado apresentou hiperplasia da polpa branca em relação ao grupo eutireóideo não-ovariectomizado. Não houve diferença na expressão de CDC47 entre os grupos. Conclui-se que a hipofunção tireoidiana e gonadal apresentam efeitos distintos no baço e que na associação hipotireoidismo-hipogonadismo há aumento menos acentuado do índice apoptótico e da expressão de caspase-3 esplênica do que no hipotireoidismo isolado.
Apoptosis, proliferation and histomorphometry of spleen were investigated in ovariectomized and non-ovariectomized adult Wistar rats maintained in hypothyroidism induced by daily administration of propylthiouracil (PTU) during 120 days. Two groups ovariectomized euthyroid and non-ovariectomized euthyroid were used as controls. Plasma was collected for free T4 dosage and the spleen for histomorphometry analysis, apoptosis index and the immunohistochemistry expression of caspase 3 and CDC47. Values of free T4 were lower in rats treated with PTU (p<0.05). In the hypothyroid groups there was some decrease in the spleen weight as well as the number and size of lymphoid follicles and there was some increase in the apoptotic index and the caspase 3 expression (p<0.05). However, the increase in the apoptosis index and the expression of caspase 3 in ovariectomized hypothyroid rats spleen was less accentuated than non-ovariectomized hypothyroid ones (p<0.05). The ovariectomized euthyroid group presented white pulp hyperplasia in comparison to the non-ovariectomized euthyroid group. There was no difference in the CDC47 expression between groups. It was concluded that the thyroid and ovarian hypofunction have distinct effects on the spleen and that in the hypothyroidism-hypogonadism association, the increase in the apoptosis index and in the expression of splenic caspase 3 is not as much as in isolated hypothyroidism.
Assuntos
Animais , Feminino , Ratos , Apoptose , Proliferação de Células , Hipotireoidismo/metabolismo , Ovariectomia , Baço , Glândula Tireoide/metabolismo , Antitireóideos , Adenosina Trifosfatases/análise , Adenosina Trifosfatases/metabolismo , /análise , /metabolismo , Modelos Animais de Doenças , Proteínas de Ligação a DNA/análise , Proteínas de Ligação a DNA/metabolismo , Hipogonadismo/metabolismo , Hipotireoidismo/sangue , Hipotireoidismo/induzido quimicamente , Tecido Linfoide/metabolismo , Tecido Linfoide/patologia , Propiltiouracila , Ratos Wistar , Baço/citologia , Baço/metabolismo , Baço/patologia , Glândula Tireoide/citologia , Tiroxina/sangueRESUMO
BACKGROUND & OBJECTIVES: Cultivated limbal stem cell transplantation is being used as a current treatment modality for limbal stem cell deficiency. However, use of allogenic biological material as substrate is associated with risks of transmission of certain diseases and allograft rejection. Therefore development of non-toxic biodegradable synthetic polymers is important. We undertook this study to evaluate the use of a synthetic polymer Mebiol gel as a substrate for the growth of limbal phenotype cells and cornea phenotype cells from limbal explants. METHODS: Human cadaveric limbal explants cells were cultivated on Mebiol gel. The proliferative capacity of cultivated cells was analyzed with thymidine incorporation studies. Immunostaining for presumed limbal stem cell association markers and cornea differentiation markers was performed and confirmed with reverse transcription (RT-PCR). RESULTS: The limbal explants underwent proliferation in vitro. The cultivated cells expressed the presumed limbal stem cell association markers (ABCG2 and p63), the transient amplifying cell markers (connexin 43, integrin alpha9) and the cornea differentiation marker (K3). RT PCR confirmed the immunohistochemical data. INTERPRETATION & CONCLUSION: Our findings showed that the synthetic polymer Mebiol gel was able to support limbal explant proliferation. The cultured cells expressed presumed limbal stem cell association markers, transient amplifying cells and cornea phenotype markers. Mebiol Gel can be used as a scaffold for growing limbal explants.
Assuntos
Transportadores de Cassetes de Ligação de ATP/análise , Técnicas de Cultura de Células/métodos , Sobrevivência Celular , Proteínas de Ligação a DNA/análise , Imunofluorescência , Géis , Humanos , Imuno-Histoquímica , Cadeias alfa de Integrinas/análise , Limbo da Córnea/química , Proteínas de Neoplasias/análise , Células-Tronco/citologia , Transativadores/análise , Proteínas Supressoras de Tumor/análiseRESUMO
BACKGROUND/AIMS: In order to identify microsatellite instability (MSI), the test based on the polymerase chain reaction (PCR) can be used. However, PCR is not routinely performed in all hospital laboratories. Recently, immunohistochemistry (IHC) for MLH1 and MSH2 proteins has been reported as a rapid and useful method for MSI. However, the efficacy of IHC in the detection of the MSI has not been well established. The aim of this study was to evaluate the usefulness of IHC in the detection of the MSI by comparing it with the test results using PCR in colorectal cancer (CRC). METHODS: Paraffin-embedded normal and tumor tissues from seventy-five patients who underwent surgical resection of CRC were used. Abnormal expression of MLH1 and MSH2 protein was determined by IHC using MLH1 and MSH2 antibodies. Normal and tumor DNAs were obtained from thirty CRC tissues that showed abnormal expression of MLH1 and MSH2 proteins by IHC. The MSI status was confirmed by PCR using five markers. RESULTS: Thirty tumors showed abnormal expression of MLH1 and MSH2 proteins by IHC, but only three tumors out of them were confirmed to have MSI by PCR. CONCLUSIONS: This result suggests that IHC with MLH1 and MSH2 antibodies does not seem to be a useful method to identify MSI in CRC, therefore PCR is required for detection of the MSI.
Assuntos
Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Adaptadoras de Transdução de Sinal , Proteínas de Transporte , Neoplasias Colorretais/genética , Proteínas de Ligação a DNA/análise , Imuno-Histoquímica , Repetições de Microssatélites , Proteína 2 Homóloga a MutS , Proteínas de Neoplasias/análise , Proteínas Nucleares , Reação em Cadeia da Polimerase , Proteínas Proto-Oncogênicas/análiseRESUMO
To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.
Assuntos
Medições Luminescentes , Proteínas de Ligação a DNA/análise , Proteínas de Ligação a DNA/genética , Ensaio de Desvio de Mobilidade Eletroforética , NF-kappa B/análise , NF-kappa B/genética , NF-kappa B/metabolismo , Ratos Sprague-Dawley , Ativação Transcricional , Transativadores/análise , Transativadores/genética , Transcrição GênicaRESUMO
Autoimmune sera have been used in the diagnosis of autoimmune diseases as well as the analysis of nuclear substructures. In an attempt to study the biological characteristics of the nuclear matrix, we screened human sera using immunofluorescent staining and immunoblot. We detected antibodies against nuclear matrix (NM), a remnant nonchromatin protein compartment after the treatment of detergent, salt and nuclease, in 212 out of 284 tested sera (74.6%) by immunoblot. Peptides with molecular weights of 70 kDa, 50 kDa and 25 kDa were detected in the order of frequency. Clinical informations of 198 out of 212 cases were available and went as follows: 38 cases were autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis; 132 non-autoimmune and non-neoplastic diseases; 16 neoplastic diseases and 12 cases unclassified. The immunofluorescent staining intensity by anti-nuclear matrix protein (NMP) antibodies decreased variably, but fibrillogranular, speckled and nucleolar immunolocalization patterns were retained after in situ fractionation. Ku70 and La protein were detected by anti-NMP antibodies. Immunolocalization by anti-NMP antibodies indicates that the NMPs constitute a variety of characteristic nuclear substructures and may serve as autoantigens in diverse human diseases. In addition, the presence of Ku70 and La protein as NMPs suggests that the NM can be functionally active in association with DNA or RNA.