RESUMO
The formation of distant metastasis resulting from vascular dissemination is one of the leading causes of mortality in nonsmall cell lung cancer (NSCLC). This metastatic dissemination initiates with the adhesion of circulating cancer cells to the endothelium. The minimal requirement for the binding of leukocytes to endothelial Eselectins and subsequent transmigration is the epitope of the fucosylated glycan, sialyl Lewis x (sLex), attached to specific cell surface glycoproteins. sLex and its isomer sialyl Lewis a (sLea) have been described in NSCLC, but their functional role in cancer cell adhesion to endothelium is still poorly understood. In this study, it was hypothesised that, similarly to leukocytes, sLe glycans play a role in NSCLC cell adhesion to Eselectins. To assess this, paired tumour and normal lung tissue samples from 18 NSCLC patients were analyzed. Immunoblotting and immunohistochemistry assays demonstrated that tumour tissues exhibited significantly stronger reactivity with antisLex/sLea antibody and Eselectin chimera than normal tissues (2.2 and 1.8fold higher, respectively), as well as a higher immunoreactive score. High sLex/sLea expression was associated with bone metastasis. The overall α1,3fucosyltransferase (FUT) activity was increased in tumour tissues, along with the mRNA levels of FUT3, FUT6 and FUT7, whereas FUT4 mRNA expression was decreased. The expression of Eselectin ligands exhibited a weak but significant correlation with the FUT3/FUT4 and FUT7/FUT4 ratios. Additionally, carcinoembryonic antigen (CEA) was identified in only 8 of the 18 tumour tissues; CEApositive tissues exhibited significantly increased sLex/sLea expression. Tumour tissue areas expressing CEA also expressed sLex/sLea and showed reactivity to Eselectin. Blot rolling assays further demonstrated that CEA immunoprecipitates exhibited sustained adhesive interactions with Eselectinexpressing cells, suggesting CEA acts as a functional protein scaffold for Eselectin ligands in NSCLC. In conclusion, this work provides the first demonstration that sLex/sLea are increased in primary NSCLC due to increased α1,3FUT activity. sLex/sLea is carried by CEA and confers the ability for NSCLC cells to bind Eselectins, and is potentially associated with bone metastasis. This study contributes to identifying potential future diagnostic/prognostic biomarkers and therapeutic targets for lung cancer.
Assuntos
Antígeno Carcinoembrionário/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Selectina E/metabolismo , Fucosiltransferases/metabolismo , Neoplasias Pulmonares/metabolismo , Antígeno Sialil Lewis X/metabolismo , Idoso , Carcinoma Pulmonar de Células não Pequenas/patologia , Adesão Celular/fisiologia , Feminino , Humanos , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Metástase Neoplásica , Polissacarídeos/metabolismoRESUMO
BACKGROUND: The E-selectin ligands expressed by cancer cells mediate adhesion of circulating cancer cells to endothelial cells, as well as within tissue microenvironments important for tumor progression and metastasis. The identification of E-selectin ligands within cancer tissue could yield new biomarkers for patient stratification and aid in identifying novel therapeutic targets. The determinants of selectin ligands consist of sialylated tetrasaccharides, the sialyl Lewis X and A (sLeX and sLeA), displayed on protein or lipid scaffolds. Standardized procedures for immunohistochemistry make use of the antibodies against sLeX and/or sLeA. However, antibody binding does not define E-selectin binding activity. METHODS: In this study, we developed an immunohistochemical staining technique, using E-selectin-human Ig Fc chimera (E-Ig) to characterize the expression and localization of E-selectin binding sites on paraffin-embedded sections of different cancer tissue. RESULTS: E-Ig successfully stained cancer cells with high specificity. The E-Ig staining show high reactivity scores in colon and lung adenocarcinoma and moderate reactivity in triple negative breast cancer. Compared with reactivity of antibody against sLeX/A, the E-Ig staining presented higher specificity to cancer tissue with better defined borders and less background. CONCLUSIONS: The E-Ig staining technique allows the qualitative and semi-quantitative analysis of E-selectin binding activity on cancer cells. The development of accurate techniques for detection of selectin ligands may contribute to better diagnostic and better understanding of the molecular basis of tumor progression and metastasis.
Assuntos
Selectina E/metabolismo , Ligantes , Neoplasias/metabolismo , Biomarcadores , Selectina E/genética , Humanos , Imuno-Histoquímica , Neoplasias/genética , Neoplasias/patologia , Inclusão em ParafinaRESUMO
A brucelose na espécie ovina tem recebido destaque, uma vez que se trata de uma enfermidade que acomete o sistema reprodutivo dos animais, provocando sério comprometimento no setor produtivo. Dessa forma, objetivou-se a avaliação de três métodos para o diagnóstico da brucelose ovina: o ensaio imunoenzimático indireto (ELISAi), a técnica imunodifusão em gel de ágar (IDGA) e a reação em cadeia da polimerase (PCR). Para tanto, utilizaram-se 211 amostras de sangue de ovinos oriundos de propriedades de nove municípios da microrregião homogênea de Teresina, Piauí. As 211 amostras de sangue foram submetidas aos testes sorológicos e à PCR, visando detectar anticorpos anti-B. ovis e DNA de Brucella ovis, respectivamente. Foram obtidos resultados positivos nos testes sorológicos, sendo 36 (17,06%) positivos no teste IDGA e sete (3,31%) positivos no teste ELISAi, contudo não houve resultados positivos na técnica de PCR. Dos métodos de diagnóstico utilizados neste estudo, o teste IDGA foi o que apresentou melhor desempenho na detecção de animais reagentes, quando comparado ao teste ELISAi e à PCR em amostras de sangue, e o percentual de animais soropositivos sugere uma ampla distribuição de ovinos infectados por Brucella ovis na região em estudo, o que pode causar prejuízos aos produtores.(AU)
Brucellosis in sheep has received a major focus, since it is a disease that affects the reproductive system of animals, causing serious impairment in the productive sector. Thus, three methods for the diagnosis of ovine brucellosis were evaluated as goal, the indirect Linked Immunosorbent Assay (ELISAi) test, the Immunodiffusion Agar Gel (AGID) technique and the Polymerase Chain Reaction (PCR). Therefore, we used 211 sheep blood samples from properties of nine municipalities of the homogeneous micro-region of Teresina, Piaui. The 211 blood samples were subjected to serologic testing and PCR to detect anti-B. ovis antibodies, and Brucella ovis DNA, respectively. Positive results in serological tests were obtained, 36 (17%) positive in the AGID test and seven (3.3%) positive to the ELISAi test, however, there were no positive results in the PCR technique. Of the diagnostic methods used in this study, the AGID test was the one that presented the best performance in the detection of reactive animals, when compared to ELISAi and PCR in blood samples and, the percentage of seropositive animals suggests a wide distribution of Brucella ovis infected sheep in the study region and could cause loss to producers.(AU)
Assuntos
Animais , Brucelose Bovina/diagnóstico , Imunodifusão/estatística & dados numéricos , Técnicas Imunoenzimáticas/estatística & dados numéricos , Reação em Cadeia da Polimerase/estatística & dados numéricos , SorologiaRESUMO
Pompe disease is a rare autosomal recessive neuromuscular disorder caused by acid α-glucosidase enzyme (GAA) deficiency and divided into two distinct variants, infantile- and late-onset. The late-onset variant is characterized by a spectrum of phenotypic variation that may range from asymptomatic, to reduced muscle strength and/or diaphragmatic paralysis. Since muscle strength loss is characteristic of several different conditions, which may also cause diaphragmatic paralysis, a protocol was created to search for the diagnosis of Pompe disease and exclude other possible causes. METHODS: We collected a sample size of 18 patients (10 females, 8 males) with a median age of 60 years and diagnosis of diaphragmatic paralysis of unknown etiology, followed in the Pulmonology outpatient consultation of 9 centers in Portugal, over a 24-month study period. We evaluated data from patient's clinical and demographic characteristics as well as complementary diagnostic tests including blood tests, imaging, neurophysiologic and respiratory function evaluation. All patients were evaluated for GAA activity with DBS (dried blood test) or serum quantification and positive results confirmed by serum quantification and sequencing. RESULTS: Three patients were diagnosed with Pompe's disease and recommended for enzyme replacement therapy. The prevalence of Pompe, a rare disease, in our diaphragmatic paralysis patient sample was 16.8%. CONCLUSION: We conclude that DBS test for GAA activity should be recommended for all patients with diaphragmatic paralysis which, despite looking at all the most common causes, remains of unknown etiology; this would improve both the timing and accuracy of diagnosis for Pompe disease in this patient population. Accurate diagnosis will lead to improved care for this rare, progressively debilitating but treatable neuromuscular disease.
Assuntos
Doença de Depósito de Glicogênio Tipo II/epidemiologia , Doença de Depósito de Glicogênio Tipo II/etiologia , Paralisia Respiratória/complicações , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Epidemiológicos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Portugal/epidemiologia , PrevalênciaRESUMO
Este trabalho teve como objetivo relatar a pesquisa de sorovares de leptospiras em animais de uma fazenda localizada no Município de Buriti dos Lopes, integrante da Bacia Leiteira de Parnaíba, Piauí, Brasil. Foram colhidas amostras de sangue de 18 vacas da raça Girolando que abortaram no período de julho de 2004 a abril de 2005. As amostras foram submetidas ao teste de soroaglutinação microscópica (SAM) utilizando-se antígenos vivos de 23 sorovares de leptospiras patogênicas. 83,33% (15/18) das amostras apresentaram aglutininas anti-leptospira, sendo 53,33% de sorovar Hardjoprajitno (8/15), 6,67% para Hardjobovis (1/15), Wolffi (1/15) e Shermani (1/15) e quatro (4/15) co-agluitinações. A amostra do reprodutor apresentou aglutininas antileptospira para o sorovar Hardjobovis com titulação 1:400 o que sugere que este animal teve participação na disseminação da infecção no rebanho.
This study aimed to report the leptospira serovar varieties in animals from a farm located in the county of Buriti dos Lopes, within the so-called "milk basin" of Parnaíba, state of Piauí, Brazil. Samples of blood were collected from 18 Girolando cows who aborted between July 2004 and April 2005, and a sample was taken from a breeder bull. The samples were subjected to the microscopic agglutination test (SAM), using live antigens of 23 pathogenic leptospira serovars. Serum samples were screened in the dilution of 1:100. Data analysis revealed that 83.33% (15/18) of the samples showed anti-leptospira agglutinins, of which 53.33% were for the Hardjoprajitno serovar (8/15), 6.67% each for Hardjobovis (1/15), Wolffi (1/15) and Shermani (1/15), along with 4 (4/15) coagglutinations. The breeder-bull sample showed anti-leptospira agglutinins for serovar Hardjobovis with titration of 1:400, suggesting that this animal had participated in the spread of the infection in the herd.
Assuntos
Animais , Bovinos , Aborto Animal/etiologia , Leptospira , Brasil , Testes de Hemaglutinação/veterinária , Testes Sorológicos/veterináriaRESUMO
Realizou-se estudo sorológico, por meio do teste de soroaglutinação microscópica, utilizando antígenos vivos, para pesquisa de aglutininas anti-leptospiras em bovinos de leite na Microrregião de Parnaíba, PI, em dois períodos do ano: estações seca e chuvosa. Das 1975 amostras analisadas, pertinentes a 16 rebanhos, 1044 (52,9 por cento) mostraram-se positivas a pelo menos um sorovar e todos os rebanhos apresentaram animais reagentes. A prevalência variou entre 25 por cento e 81 por cento. O sorovar de maior prevalência foi Hardjo reagindo em 39,5 por cento, seguido de Wolffi com 26,7 por cento e Hebdomadis com 12,2 por cento. Os sorovares mais freqüentes foram Hardjo e Wolffi presentes em 100 por cento dos rebanhos. Foram ainda encontrados animais reagentes aos sorovares Australis, Bratislava, Autumnalis, Butembo, Icterohaemorrhagiae, Shermani e Patoc. Houve correlação positiva entre a proporção de animais sororreagentes para leptospirose e pluviometria, mas não para temperatura. Foi constatada associação entre infecção e ocorrência de transtornos reprodutivos, e o sorovar que apresentou maior associação foi o Hardjo
This study was carried out by serologic sampling (microscopic agglutination test, living antigens) for anti-leptospiral antibodies in dairy farms of Parnaíba microrregion, state of Piauí, Brazil, during dry and rainny seasons. All 16 flocks were infected and 1044 out of 1975 sera samples (52.9 percent) were positive for at least one serovar. Higher prevalence was serovar Hardjo (39.5 percent) folowed by Wolffi (26.7 percent) and Hebdomadis (12.2 percent). Hardjo and Wolffi were found in all flocks. Serovars Australis, Bratislava, Autumnalis, Butembo, Hebdomadis, Icterohaemorrhagiae, Hardjo, Wolffi, Shermani and Patoc, were also present. A positive correlation between prevalence of infection and pluviometric was detected, but temperature ratios. Association between infection and reproductive failures was evidenced mainly regarding Hardjo serovar
Assuntos
Animais , Aglutininas/isolamento & purificação , Bovinos/microbiologia , Estudos Transversais , Leptospirose/etiologia , Leptospirose/veterinária , Comportamento Sexual Animal , Testes Sorológicos/métodos , Estações do Ano/análiseRESUMO
The objetive of studing reservoir slope behaviour is not only to detect pre - existing and potential sliding areas with disastrous consequences, but also to foresee the nature and extension of constructed and natural environment damage. In the first part of the lecture the risk assessment framework and its application to landslides into reservoirs of dams is introduced, because a traditional approach does not attempt to explicitly quantify the degree of safety needed for a particular landslide (AU)