Danielle Resende Camisasca

Comparative proteomic analysis of whole saliva from chronic periodontitis patients

Chronic periodontal disease is a chronic inflammatory process affecting tooth supporting tissues in the presence of pathogenic bacterial biofilm. There is some evidence for changes in the protein composition of whole saliva from chronic periodontitis patients, but there have been no studies using a proteomic approach. Hence, the aim of this study was to compare the protein profiles of unstimulated whole saliva from patients with periodontitis and healthy subjects by two complementary approaches (2D-gel electrophoresis and liquid chromatography). Protein spots of interest were analyzed by MALDI-TOF-TOF, and the data was complemented by an ESI-Q-TOF experiment. The analyses revealed that subjects with periodontal disease have increased amounts of blood proteins (serum albumin and hemoglobin) and immunoglobulin, and they have a lower abundance of cystatin compared to the control group. A higher number of protein spots were observed in the periodontitis group, of which most were identified as alpha-amylase. This higher number of alpha-amylase variants seems to be caused by hydrolysis by cysteine proteases under such inflammatory conditions. This approach gives novel insights into alterations of salivary protein in presence of periodontal inflammation and may contribute to the improvement of periodontal diagnosis.

Expression of Bcl-2 family proteins and associated clinicopathologic factors predict survival outcome in patients with oral squamous cell carcinoma.

The aims of this study were to assess the expression levels of three proteins involved in apoptosis--Bcl-2, Bcl-X, and Bax--and evaluate their relationship with clinicopathologic features and survival in oral squamous cell carcinoma (OSCC). Immunohistochemistry was used to evaluate protein expression in 53 primary OSCCs treated by radical surgery with free margins at a single institution in 1999. Histologic specimens were graded and analyzed for perineural invasion, lymphocytic infiltrate, and pattern of invasion. Digital image analysis was performed to quantify immunostaining. Survival was analyzed using the Kaplan-Meier method and Coxs proportional hazard model. Cancer-specific 5-year survival (CSS) was 61% (56% overall survival (OS), and 51% disease-free interval (DFI)). Kaplan-Meier analysis identified pathologic stage (p=0.0007, log-rank test, OS), negative nodes status (pN) (p<0.0001, log-rank test, OS), presence of lymphocytic infiltrate (p=0.0084, log-rank test, OS), and high Bax expression (p=0.025, log-rank test, OS) to each be associated with both better OS and CSS. Tongue tumors (p=0.0179, log-rank test), worst pattern of invasion (p=0.0293, log-rank test), lack of lymphocytic infiltrate (p=0.0328, log-rank test), perineural invasion (p=0.0448, log-rank test), poorly differentiated tumors (p=0.0318, log-rank test), and low Bcl-X expression (p=0.044, log-rank test) were all associated with a low DFI. Cox regression found pN, lymphocytic infiltrate, and Bax expression to be independent prognostic factors for OS and CSS, whereas lymphocytic response and tongue tumors were predictors of DFI. Bcl-2 expression emerged as an independent marker of favorable CSS. Lymphocytic infiltrate was the most meaningful histopathologic parameter in survival analysis, whereas expression of Bcl-2 family members seems to be an important marker of a favorable prognosis in OSCC.

Oral squamous cell carcinoma grading systems – analysis of the best survival predictor

BACKGROUND The TNM system has been used for decades in an attempt to predict clinical behavior and appropriate therapy for oral squamous cell carcinomas. Histopathologic classifications can be useful as an additional predictive tool. The purpose of this study was to apply four grading systems (Multiparameter Grading System, Malignancy Grading of the Deep Invasive Margins, World Health Organization grading system, and Histologic Risk Assessment) to oral squamous cell carncinomas and evaluate each system based on clinicopathologic parameters and patient survival. METHODS The files of 53 patients diagnosed with primary oral squamous cell carcinoma at the Brazilian National Cancer Institute were evaluated. All hematoxylin and eosin-stained slides were reviewed to confirm the original diagnosis and to determine histopathologic grading. Clinicopathologic information was obtained from medical records and tumor registries. Statistical analysis was performed using Fishers exact test or the chi-square test, the Kaplan-Meier method, and the log-rank test. RESULTS The Multiparameter Grading System was statistically associated with pathologic staging (P = 0.02) and lymph node involvement (P = 0.0009). Differences in overall 5-year survival were significant for Histologic Risk Assessment (P = 0.015), pathologic staging (P = 0.001), lymph node status (P < 0.0001), and recurrence (P = 0.0001). Differences in cancer-specific 5-year survival were significant for Histologic Risk Assessment (P = 0.029), pathologic staging (P = 0.002), lymph node involvement (P < 0.0001), and recurrence (P < 0.0001). Poorly differentiated tumors were associated with the worst disease-free survival (P = 0.031) and recurrence (P = 0.043). CONCLUSION Of the grading systems evaluated, Histologic Risk Assessment demonstrated the best results for survival prediction in oral squamous cell carcinoma.

Analysis of the salivary proteome in gingivitis patients

BACKGROUND AND OBJECTIVE Gingivitis is a disease that is characterized by inflammation of the gingival tissue, which can progress to periodontitis and tooth loss. Although many studies have attempted to identify salivary proteins that are associated with the disease, this is the first study to use a proteomic approach to analyze and compare the proteomic profile of whole saliva from gingivitis patients and healthy controls. MATERIAL AND METHOD To analyze the saliva proteome, two-dimensional gel electrophoresis and liquid chromatography were used, followed by mass spectrometry. RESULTS The analyses showed that gingival inflammation was associated with increased amounts of blood proteins (serum albumin and hemoglobin), immunoglobulin peptides and keratins. In the control group, salivary cystatins, which were detected using capillary Liquid Chromatography on line to electrospray ionization Quadrupole Time-of-flight mass spectrometry, appeared to be more abundant. CONCLUSION This approach provides novel insight into profiles of the salivary proteome during gingival inflammation, which may contribute to improvements in diagnosis.

Oral Squamous Cell Carcinoma: Clinicopathological Features in Patients with and without Recurrence

Aim: To compare the clinicopathological profile of oral squamous cell carcinoma (OSCC) in groups with and without recurrence. Methods: Records of all patients who underwent surgery for primary OSCC at a single institution during 1999 were identified. Patient demographics, lesion site, clinical and pathologic stage, pathologic grading, pattern of invasion, lymphocytic infiltrate, perineural invasion, and treatment and survival data were collected. Descriptive statistics were calculated for each variable and survival was calculated using Kaplan-Meier and Cox models. Patients were divided into 2 groups: with (n = 25) and without (n = 28) recurrence. Results: Tongue (p = 0.02) and poorly differentiated (p = 0.04) tumors were associated with recurrence. Kaplan-Meier and Cox models revealed tobacco use and the absence of lymphocytic infiltrate to be associated with the poorest survival in recurrent OSCC. Conclusion: The tumor site, tobacco use, and pathological features were involved in the recurrence of OSCC and should be taken into account for OSCC treatment and follow-up.

Reproducibility of Immunostaining Quantification and Description of a New Digital Image Processing Procedure for Quantitative Evaluation of Immunohistochemistry in Pathology

Quantification of immunostaining is a widely used technique in pathology. Nonetheless, techniques that rely on human vision are prone to inter- and intraobserver variability, and they are tedious and time consuming. Digital image analysis (DIA), now available in a variety of platforms, improves quantification performance: however, the stability of these different DIA systems is largely unknown. Here, we describe a method to measure the reproducibility of DIA systems. In addition, we describe a new image-processing strategy for quantitative evaluation of immunostained tissue sections using DAB/hematoxylin-stained slides. This approach is based on image subtraction, using a blue low pass filter in the optical train, followed by digital contrast and brightness enhancement. Results showed that our DIA system yields stable counts, and that this method can be used to evaluate the performance of DIA systems. The new image-processing approach creates an image that aids both human visual observation and DIA systems in assessing immunostained slides, delivers a quantitative performance similar to that of bright field imaging, gives thresholds with smaller ranges, and allows the segmentation of strongly immunostained areas, all resulting in a higher probability of representing specific staining. We believe that our approach offers important advantages to immunostaining quantification in pathology.

Análise de sobrevida global em pacientes diagnosticados com carcinoma de células escamosas de boca no INCA no ano de 1999

O carcinoma de celulas escamosas de boca compreende cerca de 90 a 95% de todas as neoplasias malignas da boca e e um dos tipos de câncer mais frequentes no Brasil. O indice de sobrevida em 5 anos e baixo e permaneceu estavel nas ultimas decadas, apesar dos avancos nas terapias. O objetivo deste estudo foi analisar o perfil e a sobrevida global dos pacientes diagnosticados com carcinoma de celulas escamosas de boca no ano de 1999 no Instituto Nacional de Câncer. Dos 320 pacientes incluidos no estudo, 79,4% eram homens. A idade media foi de 56,7 anos, e 82,2% deles fumavam e/ou bebiam. A lingua, seguida do assoalho de boca foram os locais mais acometidos. A maioria (68,9%) dos pacientes foi diagnosticada em estadios tardios e submetida a radioterapia exclusiva (53,6%). A sobrevida media no periodo do estudo foi de 29,4 meses. Os pacientes dos estadios iniciais apresentaram maior sobrevida, assim como aqueles submetidos apenas a cirurgia como forma de tratamento e os que nao apresentaram linfonodos acometidos ao diagnostico. Tumores localizados em palato duro e mucosa jugal apresentaram pior prognostico. Foram fatores preditivos independentes de melhor sobrevida os tumores T1 ou T2 (p=0,001), sem acometimento de linfonodos (p=0,012) e nao localizados em mucosa jugal (p=0,021). O diagnostico do câncer oral ainda se faz em estadios tardios, o que influencia negativamente a sobrevida global dos pacientes. Maior enfase deve ser dada a capacitacao dos profissionais para o reconhecimento precoce do câncer e a conscientizacao da populacao de risco.

Plaquetas: ainda um alvo terapêutico

As plaquetas sao fragmentos citoplasmaticos anucleados presentes no sangue e produzidos na medula ossea a partir dos megacariocitos. O objetivo deste trabalho e rever as bases mecanisticas e moleculares das plaquetas, revelando sua participacao em sindromes importantes e na trombose arterial, alem de seu potencial como alvo terapeutico para o desenho de novos agentes antitromboticos.

Recurrent bilateral gingival peripheral calcifying epithelial odontogenic tumor (Pindborg tumor): A case report

Calcifying epithelial odontogenic tumor (CEOT) is an extremely rare, benign neoplasm, accounting for approximately 1% of all odontogenic tumors. Peripheral CEOTs commonly resemble oral hyperplastic or reactive lesions and are histologically similar to their intraosseous counterparts. We report an unusual case of multifocal peripheral CEOT. A 40-year-old female presented with bilateral soft, painful, erythematous, gingival swellings localized in premolar areas of the mandibular gingiva. The presumptive diagnosis was bilateral pyogenic granuloma. The masses were surgically excised under local anesthesia without bone curettage and both recurred 12 months later. Morphologic features, and histochemical and immunohistochemical tests revealed bilateral peripheral calcifying odontogenic epithelial tumor. There is no clinical or radiographic evidence of recurrence 3.5 years after excision. This multifocal phenomenon has been reported previously only for intraosseous CEOT. Gingival masses must be carefully evaluated for clinical and histologic evidence of neoplasia.

Usefulness of tissue autofluorescence imaging in actinic cheilitis diagnosis

Abstract. Actinic cheilitis (AC) is a potentially malignant disorder of the lips. Because of its heterogeneous clinical aspect, it is difficult to indicate representative biopsy area. The purpose of this study was to evaluate the usefulness of tissue autofluorescence in AC diagnosis. The system was composed of a 405-nm light-emitting diode, sent to the sample by a dichroic, that allows the fluorescence signal to reach a camera directly plugged in the system. Fifty-seven patients with clinical diagnosis of AC and 45 normal volunteers were selected. According to clinical and fluorescence features, one or more areas were selected for biopsies in the AC group and epithelial dysplasia (ED) grades were established. The autofluorescence images were processed by a clustering algorithm for AC automated diagnosis. The tissue autofluorescence image revealed a heterogeneous pattern of loss and increase of fluorescence in patients with AC. ED was found in 93% of the cases, and most of the areas graded as moderate or severe ED were chosen with the aid of autofluorescence. The processed autofluorescence images from AC patients showed a higher number of spots in an irregular pattern. Tissue autofluorescence image system is a useful technique in association with clinical examination for AC diagnosis.