DETECTION OF INFECTIOUS-DISEASE AGENTS IN TISSUE BY IMMUNOCYTOCHEMISTRY

1. Immunocytochemical procedures have played an increasingly larger role in the identification of infectious disease agents in tissue sections owing to the increased availability and specificity of antibody reagents, the great sensitivity of the methods, and the relative facility with which the studies are performed.2. Immunocytochemical methods can be applied to routine formalin-fixed tissue for the detection of infectious agents such as viruses, bacteria, fungi, and protozoa among other microorganisms for diagnostic and research purposes.

Experimental alcoholism and pathogenesis of prostatic diseases in UChB rats

Previous studies have shown that long-term alcohol treatment has negative effects on prostatic stromal-epithelial interaction. Thus, the aim of the present study was to analyze the histochemical, immunohistochemical and ultrastructural alterations that occur in the prostatic stroma and epithelium of rats submitted to chronic alcohol ingestion and alcohol abstinence, as well as to establish the relationship between these changes and prostatic diseases. Thirty male rats (10 Wistar and 20 UChB rats) were divided into three experimental groups: the control group received tap water, the alcoholic group received ethanol diluted to 10 degrees G.L. for 150 days, and the abstinent group received the same liquid diet as the alcoholic group up to 120 days of treatment and only tap water for 30 days thereafter. At the end of treatment, all animals were sacrificed and the ventral lobe of the prostate was removed and processed for histochemical, immunohistochemical and ultrastructural analyses. In addition, plasma testosterone levels were measured. The results showed, prostatic intraepithelial neoplasia, infolding of the epithelium towards the stroma, stromal hypertrophy and the presence of inflammatory cells in alcoholic animals. In the abstinent group, alterations were noted mainly in the stromal area. In conclusion, ethanol triggers alterations in prostatic epithelial and stromal compartments, affecting the stromal microenvironment and predisposing the organ to pathological processes. (C) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

Reproducibility and validity of self-perceived oral health conditions

The reproducibility and validity of self-perceived periodontal, dental, and temporomandibular joint (TMJ) conditions were investigated. A questionnaire was applied in interview to 200 adults aged from 35 to 44, who were attending as casual patients at Araraquara School of Dentistry, Sauo Paulo State University, Sauo Paulo, Brazil. Clinical examination was based on the guidelines of the World Health Organization manual. The interview and the clinical examination were performed in two occasions, by a calibrated examiner. Reproducibility and validity were, respectively, verified by kappa statistics (kappa) and sensitivity (Sen) and specificity (Spec) values, having clinical examination as the validation criterion. The results showed an almost perfect agreement for self-perceived TMJ (kappa=0.85) and periodontal conditions (kappa=0.81), and it was substantial for dental condition (kappa=0.69). Reproducibility according to clinical examination showed good results (kappa=0.73 for CPI index, kappa=0.96 for dental caries, and kappa=0.74 for TMJ conditions). Sensitivity and specificity values were higher for self-perceived dental (Sen=0.84, Spec=1.0) and TMJ conditions (Sen=1.0, Spec=0.8). With regard to periodontal condition, specificity was low (0.43), although sensitivity was very high (1.0). Self-perceived oral health was reliable for the examined conditions. Validity was good to detect dental conditions and TMJ disorders, and it was more sensitive than specific to detect the presence of periodontal disease.

DETECTION OF CELLULAR-IMMUNITY WITH THE SOLUBLE-ANTIGEN OF THE FUNGUS SPOROTHRIX-SCHENCKII IN THE SYSTEMIC FORM OF THE DISEASE

Sporothrix schenckii is the etiologic agent of sporotrichosis, a mycosis of world-wide distribution more commonly occurring in tropical regions. The immunological mechanisms involved in the prevention and control of sporotrichosis are not fully understood but apparently include both the humoral and cellular responses. In the present investigation, cellular immunity was evaluated by in vivo and in vitro tests in mice infected with yeast-like forms of S. schenckii. The disease developed systemically and cellular immunity was evaluated for a period of 10 weeks. The soluble antigen utilized in the tests was prepared from yeast form of the fungus through the sonication (20 min: 10 sonications at 50 W at 2-min intervals). Delayed hypersensitivity and lymphocyte transformation tests showed that the cellular immune response was depressed between the 4th and 6th week of infection when the animals were challenged with the soluble fungal antigen. This depression frequently indicates worsening of the disease, with greater involvement of the host. This is a promising field of research for a better understanding of the pathogeny of this mycosis.

COX-2 inhibition decreases VEGF expression and alveolar bone loss during the progression of experimental periodontitis in rats

Background: Vascular endothelial growth factor (VEGF) is a macromolecule of importance in inflammation that has been implicated in periodontitis. The aims of this study were to investigate VEGF expression during the progression of periodontal disease and to evaluate the effect of a preferential cyclooxygenase (COX)-2 inhibitor meloxicam on VEGF expression and alveolar bone loss in experimentally induced periodontitis.Methods: A total of 120 Wistar rats were randomly separated into groups 1 (control) and 2 (meloxicam, 3 mg/kg/day, intraperitoneally, for 3, 7, 14, or 30 days). Silk ligatures were placed at the gingival margin level of the lower right first molar of all rats. VEGF expression was assessed by reverse transcription-polymerase chain reaction (RT-PCR), Western blot (WB), and immunohistochemical (IHC) analyses. The hemiarcades were processed for histopathologic analysis. RT-PCR and WB results were submitted to analysis of variance, the Tukey test, and Pearson correlation analysis (P<0.05).Results: A reduction in alveolar bone resorption was observed in the meloxicam-treated group compared to the control group at all periods studied. There was a positive correlation between COX-2 mRNA and VEGF mRNA in the gingival tissues and periodontal disease (R = 0.80; P = 0.026). Meloxicam significantly reduced the increased mRNA VEGF expression in diseased tissues after 14 days of treatment (P = 0.023). Some alterations in VEGF receptor I mRNA expression were observed, but these were not statistically significant. VEGF protein expression in WB experiments was significantly higher in diseased sites compared to healthy sites (P<0.05). After 14 days of treatment with meloxicam, an important decrease in VEGF protein expression was detected in diseased tissues (P = 0.08). Qualitative IHC analysis revealed that VEGF protein expression was higher in diseased tissues and decreased in tissues from rats treated with meloxicam.Conclusions: The present data suggest an important role for VEGF in the progression of periodontal disease. Systemic therapy with meloxicam can modify the progression of experimentally induced periodontitis in rats by reducing VEGF expression and alveolar bone loss.

TESA-blot for the diagnosis of Chagas disease in dogs from co-endemic regions for Trypanosoma cruzi, Trypanosoma evansi and Leishmania chagasi

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Serology of paracoccidioidomycosis. II. Correlation between class-specific antibodies and clinical forms of the disease

Untreated and previously treated patients with paracoccidioidomycosis were studied for: (i) serum levels of total IgG, IgM and IgA immunoglobulins, by radial immunodiffusion and Paracoccidioides brasiliensis (Pb) antibodies, by indirect immunofluorescence; (ii) correlation between their levels with the clinical forms of the disease; (iii) correlation between the serum titres obtained by tube precipitin with those of anti-Pb IgG, IgM and IgA. In the untreated group, serum IgG levels were significantly increased in patients with the more systemic forms of the disease, especially the acute progressive form. Serum IgA levels were significantly increased in all patients with no statistical difference between clinical forms. Serum IgM levels were normal in all patients. Anti-Pb IgG, IgA and IgM were detected in 97·5%, 32·5% and 45·0% of all cases, respectively. There was a sharp tendency towards higher levels of anti-Pb IgG among those with the acute progressive form (83·4%) in relation to the chronic, more localized forms, mixed form (68·0%) and isolated organic form (55·5%). In the untreated and previously treated group sera, there was positive correlation between the level of anti-Pb IgG and positivity for the tube precipitin test, suggesting that the precipitin-type antibodies are of the IgG class. Broadly, the present data demonstrate a polyclonal activation of the humoral immune system in paracoccidioidomycosis, with a positive relationship between serological results and severity of the disease. © 1984.

Estreptomicina aumenta a aderência em células epiteliais de Bacteroides melaninogenicus associado às lesões peridentárias da cara inchada dos bovinos

Cara inchada of cattle (CI) is a periodontitis which occurs in calves in several regions of Brazil. Its progressive lesions, caused mainly by Bacteroides melaninogenicus and Actinomyces pyogenes, could lead to loss of teeth, malnutrition and possibly death of affected animals. For further analysis of the pathogenesis of this disease the influence of streptomycin on the adherence of the causative bacteria to oral epithelial cells of cattle was determined. Production of streptomycin by soil bacteria (Actinomycetes) in Brazil had been suggested by several authors. In our studies, streptomycin in subinibitory concentrations increased tenfold the rate of adherence with B. melaninogenicus (from 0.4 to 4.0 bacteria per bovine epithelial cell). On the contrary, streptomycin did not influence significantly the adherence of A. pyogenes nor streptococci of the viridans group. Thus, streptomycin could be a contributing factor to the development of the periodontal infections with B. melaninogenicus.

Sharpening of periodontal instruments with different sharpening stones and its influence upon root debridement--scanning electronic microscopy assessment

The objective of this study was to evaluate, through scanning electronic microscopy, the effect of sharpening with different sharpening stones on the cutting angle of periodontal curettes (Gracey 5-6), and the influence on root surfaces after debridement and planing. The experimental model consisted of two different phases. In the first, the cutting angles of fifteen stainless steel Gracey 5-6 curettes were analyzed under a scanning electronic microscope after being sharpened with different types of stones. In the second phase, the root surfaces of 25 newly extracted teeth were evaluated with a scanning electronic microscope after being debrided with curettes sharpened with different stones. Analysis of the results showed that the synthetic stones (aluminum oxide and carborundum) are more abrasive and produce more irregular cutting angles, whereas Arkansas stones are less abrasive and produce smoother and more defined cutting angles. There was no significant statistical differences among the five groups tested with regard to the degree of irregularity of the root surfaces after instrumentation.