Adriana Maria da Silva Telles

Altered lectin-binding sites in normal colon and ulcerative colitis

O presente trabalho objetivou, atraves de histoquimica com lectinas e analise digital de imagens, avaliar a expressao de carboidratos em amostras de colo normal e com colite ulcerativa. A partir de fragmentos de mucosa intestinal foram obtidos cortes histologicos (4mm) que foram incubados com lectinas (Con A, WGA, LTA e PNA), e os resultados das marcacoes foram avaliados atraves de microscopia optica e sistema de analise de imagens. Os resultados obtidos revelaram uma intensa marcacao para as celulas inflamatorias, principalmente neutrofilos infiltrados no tecido de reto e sigmoide, bem como celulas das glândulas intestinais. As lectinas WGA e LTA exibiram padroes distintos de marcacao entre o epitelio normal e os casos de colite ulcerativa. As lectinas PNA e Con A falharam em reconhecer os carboidratos celulares nos casos estudados em ambos os grupos. Os resultados obtidos foram confirmados pela analise de imagem. As observacoes obtidas sugerem que as lectinas WGA e LTA sao marcadores promissores para diferenciar o epitelio normal do padrao inflamatorio da colite ulcerativa, indicando uma expressao distinta de N-acetilglicosamina e L-fucose nos respectivos casos estudados.

Immunoelectrophoretic study on common antigens of São Lourenço da Mata and Belo Horizonte strains of Schistosoma mansoni adult worms and Biomphalaria snails

Immunoelectrophoretic studies on common antigens were carried out by using rabbits sera immunized against São Lourenço da Mata and Belo Horizonte strains of Schistosoma mansoni adult worms and antigens of Biomphalaria glabrata pigmented (Jaboatão--PE); B. glabrata albino (Belo Horizonte--MG) and B. straminea (São Lourenço da Mata, PE). Furthermore, the reverse approach was proceeded, namely, sera anti Biomphalaria snails produced in rabbits were tested against both strains of Schistosoma adult worm antigens. The analysis of the common antigens between the SLM strains of S. mansoni adult worm and B. glabrata pigmented showed 8 to 9 precipitin bands, 3 bands with B. glabrata albino and only 1 band with B. straminea crude extracts. On the other hand, the BH strain of S. mansoni adult worm antisera produced 6 to 7 bands with B. glabrata pigmented, 5 bands with B. glabrata albino and 1 band with B. straminea antigenic extract. Biomphalaria snails crude extracts were fractionated by Sephadex G-100 column and three fractions were collected from each snail strain. The fractions were tested with anti SLM and BH strains of S. mansoni adult worm sera by immunoelectrophoresis. The common antigens fractionated from Biomphalaria snails crude extracts and those found for both strains of S. mansoni adult worm mostly existed in the first fraction and they were estimated to have molecular weight over 158,000 daltons. In our laboratory, it was found a relationship between the antigenic similarities and experimental infection rates of S. mansoni towards Biomphalaria snails so that more bands were seen with increasing infection rates of S. mansoni.

Tumour necrosis factor‐alpha (‐308G/A) promoter polymorphism is associated with ulcerative colitis in Brazilian patients

Inflammatory bowel disease consists of multifactorial diseases whose common manifestation is inflammation of the gastrointestinal tract and their pathogenesis remains unknown. This study aimed to analyse the gene polymorphisms in Brazilian patients with inflammatory bowel disease. A total of 101 patients diagnosed with inflammatory bowel disease were analysed for the tumour necrosis factor‐alpha (‐308 G/A; rs1800629) and interleukin‐10 (‐1082 G/A; rs1800896) gene polymorphisms. Genotyping was performed through polymerase chain reaction–sequence‐specific primer, then fractionated on 2% agarose gel and visualized after staining by ethidium bromide. The anatomic–clinical form of Crohns disease (CD) predominant was the inflammatory (32.75%), followed by fistulizing (29.31%) and 27.58% stricturing. As control group, a total of 136 healthy subjects, from the same geographical region, were enrolled. The statistical analyses were performed using R program. The frequency of the A allele at tumour necrosis factor‐alpha was high in ulcerative colitis (UC) patients (51%) than in controls (22%; P > 0.01). No statistical difference was found with the genotypic and allelic frequencies of CD patients compared to controls (P = 0.54). The polymorphism ‐1082G/A of interleukin‐10 was not statistical different between the diseases compared to controls. Tumour necrosis factor‐alpha (TNF‐α) (‐308G/A) is associated with UC onset, suggesting that the presence of ‐308A allele could confer a relative risk of 3.62 more to develop UC in general population. Further studies, increasing the number of individuals, should be performed to ratify the role of TNF‐α in the inflammatory bowel disease pathogenesis.

Análise histológica e histoquímica de fatores prognósticos em pacientes com retocolite ulcerativa

OBJECTIVES: Aiming to offer a diagnosis method or a prognostic marker, galectin-3 appears as a potential marker of the physiopathology of diverse diseases and some types of cancer, between them the colorectal cancer. METHODS: In our study, the immunohistochemical and histological profiles had been taken from intestinal biopsies of patients with ulcerative colitis (n=20) and its respective normal counterpart, from both males and females and average age of 55 years-old. The biopsies had been submitted to Periodic Acid of Schiff (PAS) for visualization of glycosaminoglycans inclusions, and Massons tricromic for visualization of collagen fibers. Fragments (4µm) had been submitted to the technique of immunohistochemistry for galectin-3 protein. The profiles of tecidual coloration had been analyzed through a work station contend an optic microscope copulated with a digital camera, both connected a computer contend software OPTIMAS®. RESULTS: Our results demonstrate a reduction of galectin-3 expression in ulcerative colitis inflammatory site from surgically and clinically treated patients. The goblet cells number from the surgically treated patients had suffered a significant reduction (p<0.022), however with increase of its glandular content. CONCLUSIONS: The data suggest that the glandular activity together with the reduction of the galectin-3 expression can assist the accompaniment of these patients.

Experimental schistosomiasis in the Common Marmoset Callithrix jacchus

In order to evaluate Callithrix jacchus as an animal model for mansoni schistosomiasis, a group of 10 male animals were once percutaneously exposed to 250 cercariae of the Schistosoma mansoni SLM (São Lourenço da Mata) strain. Animals were periodically bled for measuring serum level of enzymes and proteins and for blood cell counting. When comparing pre-infection to post-infection values, a significant increase was found for alkaline phosphatase at 15 to 120 days p.i., differential counts of eosinophil at 45 and 60 days, and total protein and global eosinophil counts at 120 days. No Schistosoma mansoni eggs were found in stools. Adult worms of small size were recovered from five animals. At day 120, the number of Schistosoma mansoni eggs/g of tissue was 0-289.7 (liver), 0-30.1 (large intestine) and 0-171.4 (small intestine). These findings lead us to classify Callithrix jacchus as a non-permissive host to the SLM strain of Schistosoma mansoni.

Histomorfometria e índice proliferativo (Ki-67) no carcinoma escamocelular in situ de pregas vocais

INTRODUCTION: Laryngeal dysplasias, precursors of squamous cell carcinomas, are uncommon lesions, inasmuch as most cases are diagnosed as invasive carcinomas. OBJECTIVE: To characterize severe dysplasia/in situ carcinoma of the vocal cords by comparing the area of involved epithelium, nuclear diameter, and proliferative index with transition and normal squamous epithelia. MATERIAL AND METHOD: Among 1,400 surgical and biopsy specimens of laryngeal lesions (1994-2006), five cases (0.35%) were selected and compared according to age with two controls with normal epithelium. Furthermore, all of them were identified and submitted to morphometric and immunohistochemical analysis (Ki-67). RESULTS: After comparing dysplastic with transition and normal squamous epithelia, it was observed an increased nuclear diameter in dysplastic epithelium. Additionally, there was no significant difference in nuclear diameter per layer between dysplastic and transition epithelia. Moreover, the proliferative index was higher in dysplastic epithelium with stained nuclei in all layers and lower in normal and transition epithelia with nuclei concentrated on the basal and parabasal layers. CONCLUSION: Dysplastic epithelium occupies larger areas. Nuclear diameter is similar in dysplastic and transition epithelia. However, the latter has uniform nuclei, in contrast with pleomorphism in dysplastic epithelium. The proliferative index (Ki-67) contributes to the differential diagnosis of intraepithelial lesions of the vocal cords, inasmuch as in severe dysplasia/in situ carcinoma there is a higher number of stained nuclei, which are seen in all epithelial layers.

Perfil imunoistoquímico das proteínas p53 e Ki67 em pacientes com retocolite ulcerativa inespecífica

Objectives: The present study evaluated, trough immunohistochemistry and histopatologic analysis, the main alterations in intestinal mucous from patients with unspecific ulcerative colitis (IUC) clinically (n=30) or surgically treated (n=30) with ileal J-pouch and permanent ileostomy. Methods: Thus, tissue slices from intestinal mucous were submitted to imunostains for p53 and Ki67 proteins, and Haematoxilin-eosin staining for comparative histopathological evaluation. Results: The results indicated important differences in inflammatory profile and erosion/necrosis sites on the both investigated groups. Related to imunoexpression was observed a higher reactivity with nuclear pattern, mainly in inflammatory tissues from IUC patients clinically treated. Conclusions: Those results showed that, according to histopathological and imunohistochemical parameters, the colonic mucous from IUC patients submitted to surgical treatment exhibited less alterations than IUC patients clinically treated. The data suggest that those methods could be helpful to diagnosis and assist the accompaniment of patients under different therapeutic routines.