Peripheral giant cell granuloma. An immunohistochemical and ultrastructural study.

OBJECTIVE: To study the nature of multinucleated and mononuclear cells from peripheral giant cell granuloma (PGCG). MATERIALS AND METHODS: Formalin-fixed, paraffin-embedded sections of 40 cases of PGCG were immunohistochemically stained for vimentin, alpha I-antichymotrypsin, CD68, S-100 protein, lysozyme, leucocyte common antigen (LCA), factor VIII-related antigen and muscle cell actin. Six cases of PGCG were also studied by transmission electron microscopy. RESULTS: Vimentin, alpha I-antichymotrypsin and CD68 were expressed in both the mononuclear and multinucleated giant cells. Dendritic mononuclear cells, positive for S-100 protein, were noted in 67.5% of the lesions, whereas lysozyme and leucocyte common antigen were detected in occasional mononuclear cells. Ultrastructural examination showed mononuclear cells with signs of phagocytosis and sometimes interdigitations with similar cells. Others presented non-specific characteristics and the third type exhibited cytoplasmic processes and occasional Birbeck granules. Some multinucleated giant cells showed oval nuclei, abundant mitochondria and granular endoplasmic reticulum whereas others presented with irregular nuclei and a great number of cytoplasmic vacuoles. CONCLUSIONS: Immunohistochemical and ultrastructural results suggest that PGCGs of the jaws are composed mainly of cells of the mononuclear phagocyte system and that Langerhans cells are present in two thirds of the lesions.

Pesquisa de anticorpos contra Brucella ovis em ovinos do Estado de São Paulo

Eight hundred and fifty sheep sera from 18 herds, located in 15 municipalities of the State of São Paulo, were examined by complement fixation test, indirect enzyme immunoassay and gel diffusion test for antibodies to Brucella ovis. The combination of the serological test results and the clinical and epidemiological data suggest that none of the examined animals was infected. The complement fixation and the gel diffusion tests showed good specificity, but the indirect enzyme immunoassay showed false positive results in nine of the sera.

Detection of circulating Paracoccidioides brasiliensis antigen in urine of paracoccidioidomycosis patients before and during treatment

For the diagnosis and follow-up of paracoccidioidomycosis patients undergoing therapy, we evaluated two methods (immunoblotting and competition enzyme immunoassay) for the detection of circulating antigen in urine samples. A complex pattern of reactivity was observed in the immunoblot test. Bands of 70 and 43 kDa were detected more often in urine samples from patients before treatment. The immunoblot method detected gp43 and gp70 separately or concurrently in 11 (91.7%) of 12 patients, whereas the competition enzyme immunoassay detected antigenuria in 9 (75%) of 12 patients. Both tests appeared to be highly specific (100%), considering that neither fraction detectable by immunoblotting was present in urine samples from the control group. gp43 remained present in the urine samples collected during the treatment period, with a significant decrease in reactivity in samples collected during clinical recovery and increased reactivity in samples collected during relapses. Reactivity of some bands was also detected in urine specimens from patients with 'apparent cure.' The detection of Paracoccidioides brasiliensis antigens in urine appears to be a promising method for diagnosing infection, for evaluating the efficacy of treatment, and for detecting relapse.

Quantitative cell-cycle protein expression in oral cancer assessed by computer-assisted system

The knowledge of cell-cycle control has shown that the capacity of malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth. Histologic diagnosis might be improved by a quantitative evaluation of more specific diagnosis biomarkers, which could help to precisely identify pre-malignant and malignant oral lesions. The aim of the present study is to evaluate whether computer-based quantitative assessment of p53, PCNA and Ki-67 immunohistochemical expression, could be used clinically to foresee the risk of oral malignant transformation. This retrospective study was carried out in ninety-five oral biopsies, 27 were classified as fibrous inflammatory hyperplasia, 40 as leukoplakia and 28 as oral squamous cell carcinoma. Sixteen out of the 40 leukoplakia were diagnosed as non-dysplastic leukoplakia, the other 24 being dysplastic leukoplakia, of which 50.0% were classified as moderate to severe dysplasia. Comparison of the four groups of oral tissues showed significant rises in p53 and Ki-67 positivity index, which increased steadily in the order benign, pre-malignant, and malignant. In contrast, it was not possible to relate higher PCNA levels with pre-malignant and malignant oral lesions. We therefore conclude that PCNA immunohistochemistry expression is probably an inappropriate marker to identify oral carcinogenesis, whereas joint quantitative evaluation of p53 and Ki-67, appears to be useful as a tumor marker, providing a pre-diagnostic estimate of the potential for cell-cycle deregulation of the oral proliferate status.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.

Immunohistochemistry as a fundamental tool for the differential diagnosis of polymorphous low-grade adenocarcinoma

Polymorphous low-grade adenocarcinoma (PLGA) is a subtype of salivary gland carcinoma with a marked predilection for the minor salivary glands. The diagnosis of this lesion can be a challenge in incisional biopsies because of its similarity to other salivary neoplasms and the difficulty of analyzing its infiltrative nature and neurotropism. Diagnosis can be facilitated, however, by immunohistochemistry, a technique that has become a fundamental tool to the pathologist. The purpose of this study was analyze the immunohistochemical aspects of 4 cases that were examined in the São José dos Campos School of Dentistry. The patients included 4 women, ages 32 to 57 years (mean, 50 years), who reported an intraoral painless lesion, ranging in size from 1 to 3 cm (mean, 1.75 cm) with a variable evolution period. Histologically, the lesions showed similar aspects, characterized by polymorphous growth pattern, including solid, cribriform, trabecular, papillary, and single-file arrays. The tumors showed positivity to cytokeratins (Cks) 7 and 8 and vimentin, variable expression for Ck 14, and a negative reaction to Cks 13 and 19 and muscle-specific actin. PLGA is a tumor of the salivary gland that must be distinguished from other neoplasms for therapeutic and prognostic considerations, and the panel of antibodies employed in this study was effective for its diagnosis.

Pro- and anti-inflammatory cytokines produced by human monocytes challenged in vitro with Paracoccidioides brasiliensis

Monocytes and macrophages play a central role in innate and adaptive immune response against systemic fungal infections. Imbalances in suppressor or stimulatory cytokine secretion caused by these cells may influence disease development, microorganism death, and the nature of the adaptive immune response. This study analyzed the monocyte cytokine profiles of healthy individuals challenged with high and low virulent strains of P. brasiliensis and mRNA cytokine expression kinetics by reverse transcription polymerase chain reaction (RT-PCR). Peripheral blood monocytes from healthy volunteers were cultured in vitro with and without virulent (Pb18) or low virulence (Pb265) strains from P. brasiliensis viable yeast cells. Interleukin-1 beta (IL-1β), IL-6, IL-8, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor-beta (TGF-β1) were measured in culture supernatants by enzyme immunoassay (ELISA), and mRNA cytokine expression was determined by RT-PCR at 0, 4, 8, 12, 18 and 48 hr. Both P. brasiliensis strains induced monocyte production of IL-1β, IL-6, IL-10 and TNF-α. Pb18 induced higher levels of IL-1β, IL-6, and IL-10 than Pb265. IL-8 and TGF-β1 levels were not significantly different from those cultured without stimulus. The mRNA cytokine expression was similar to supernatant cytokines measured by ELISA. In vitro monocyte challenge with virulent P. brasiliensis strain induces earlier and higher levels of pro- and anti-inflammatory cytokines than low virulence strain.

Immunohistochemical expression of HLA-DR in the conjunctiva of patients under topical prostaglandin analogs treatment

PURPOSE: Subclinical inflammation may be observed in patients using topical antiglaucomatous drugs. The objective of this study was to investigate inflammation in conjunctiva of glaucoma patients using prostaglandin analogs, by the detection of an immunogenetic marker (HLA-DR) and compare the effect of 3 different drugs: latanoprost, bimatoprost, and travoprost in the induction of this inflammation. SUBJECTS AND METHODS: Thirty-three patients with primary open-angle glaucoma were evaluated without and with prostaglandin analogs topical therapy. Imprints of conjunctival cells were obtained, fixed on glass slides, and prepared for immunohistochemical analysis. RESULTS: Before the use of prostaglandin analogs, 4 of the 33 patients evaluated presented expression of HLA-DR in the conjunctiva (mild). After 1 month on prostaglandin analog treatment, all but 1 patient presented HLA-DR staining. HLA-DR expression of these 32 patients was scored as mild (19 patients), medium (11 patients), or intense (2 patients). The differences were statistically significant both when the presence and the increased expression of HLA-DR were considered (P<0.001). When the 3 different groups were analyzed (latanoprost, bimatoprost, and travoprost) no statistically significant difference was found (P=0.27). CONCLUSIONS: The use of prostaglandin analogs eye drops provokes a subclinical inflammatory reaction, observed by HLA-DR expression, even after a short period of treatment, independently of the class of the prostaglandin analogs used. © 2009 Lippincott Williams & Wilkins, Inc.

A Bayesian approach to estimate the accuracy of in-house ELISA assay to measure rabies antibodies from compulsory vaccinated dogs and cattle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Identificação de espécies e genótipos de Cryptosporidium em bovinos leiteiros no Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fecal progestins during pregnancy and postpartum periods of captive red brocket deer (Mazama americana)

This study aimed to validate the enzyme immunoassay (EIA) for fecal progestin quantification of the species Mazama americana, define its excretion profile during periods of gestation and postpartum and determine the gestation period and resumption of postpartum ovarian activity in this species in captivity Fecal samples were collected twice a week during gestation and every day in the postpartum period, and analyzed using EIA The mean concentrations (±SEM) of fecal progestins during gestation were 2180.0 ± 299.1 ng/g in early pregnancy (week 1-11), 3271.4 ± 406.9 ng/g in middle pregnancy (week 12-22) and 5592.0 ± 1125.8 ng/g in late pregnancy (week 23-32) The gestation period determined for the species was 220.9 ± 1.2 days The concentration of progestins reached its peak prior to parturition and returned to baseline levels in 4 ± 0.31 days after parturition In the postpartum period, the mean concentrations of fecal progestins were 1564.2 ± 182.6 ng/g in the interval between parturition and resumption of ovarian activity, 469.8 ± 24.5 ng/g in the inter-luteal phase and 2401.7 ± 318.5 ng/g during the luteal phase, such that the postpartum period and the luteal phase differed from the inter-luteal phase Fecal progestin profiling permitted the detection of ovulation 26.9 ± 3.4 days after parturition in all the hinds studied and estimation of the mean duration of the estrous cycle, 21.3 ± 1.1 days Analysis established that concentrations of progestins above 3038.76 ng/g diagnosed pregnancy, a value determined from the week 12 of gestation Moreover, the quantification of fecal progestins by EIA proved to be an important tool for noninvasive endocrine monitoring and to obtain reproductive data on the species M americana in captivity © 2013 Elsevier B.V.

Lentinula edodes (Shiitake) modulates chemically induced mutagenesis by enhancing pitting

This study was undertaken to understand how Lentinula edodes modulates in vivo mutagenesis induced by alkylating agents in bone marrow and peripheral blood as described in our previous article. Male Swiss mice were pretreated for 15 consecutive days with aqueous extracts prepared from L. edodes, after which, the number of circulating blood cells, normal erythroid bone marrow cell cycling, and phagocytosis of micronucleated reticulocyte (MNRET) and activation of spleen macrophages were assessed. The results indicate that the antimutagenicity seen in bone marrow and peripheral blood is exerted by distinct compounds with different actions. The antimutagenic effect in bone marrow is exerted by compounds subject to degradation at deep-freeze storage temperature of -20 C. On the other hand, compounds responsible for antimutagenicity in peripheral blood are not subject to degradation at -20 C. The results also indicate that the antimutagenic action in peripheral blood leading to the reduction of circulating MNRET occurs in the spleen primarily through a phagocytic activity due to higher macrophage numbers and probably not due to the enhanced activation state of individual cells. © Mary Ann Liebert, Inc.

Efeito da interleucina-15 sobre a atividade fungicida, metabolismo oxidativo e produção de citocinas por monócitos humanos, infectados in vitro com Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Pressão intra-ocular, condições endoteliais e prostaglandina E2 no humor aquoso de cães com catarata senil madura ou hipermatura

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)