Computer-assisted analysis of p53 and PCNA expression in oral lesions infected with human papillomavirus

OBJECTIVE: To carry out a retrospective study to determine whether human papillomavirus (HPV) infection and immunohistochemical expression of p53 and proliferating cell nuclear antigen (PCNA) are related to the risk of oral cancer. STUDY DESIGN: Fifty-seven oral biopsies, consisting of 30 oral squamous papillomas (OSPs) and 27 oral squamous cell carcinomas (OSCCs) were tested for the presence of HPV 6/11 and 16/18 by in situ hybridization using catalyzed signal amplification and in situ hybridization. p53 And PCNA expression was analyzed by immunohistochemistry and evaluated quantitatively by image analysis. RESULTS: Nineteen of the 57 oral lesions (33.3%) were positive for HPV. HPV 6/11 was found in 6 of 30 (20%) OSPs and 1 of 27 (3.7%) OSCCs. HPV 16/18 was found in 10 of 27 (37%) OSCCs and 2 of 30 (6.7%) OSPs. Sixteen of the 19 HPV-positive cases (84.2%) were p53 negative; 5 (9%) were HPV 6/11 and 11 (19%) HPV 16/18, with an inverse correlation between the presence of HPV DNA and p53 expression (P=.017, P < .05). PCNA expression appeared in 18 (94.7%) of HPV positive cases, showing that HPV 16/18 was associated with intensity of PCNA expression and with OSCCs (P=.037, P < .05). CONCLUSION: Quantitative evaluation of p53 by image analysis showed an inverse correlation between p53 expression and HPV presence, suggesting protein degradation. Image analysis also demonstrated that PCNA expression was more intense in HPV DNA 16/18 OSCCs. These findings suggest involvement of high-risk HPV types in oral carcinogenesis.

Components of the acoustic swallowing signal: Preliminary study

Purpose: To analyze the components of the acoustic signal of swallowing using a specific software. Methods: Fourteen healthy subjects ranging in age from 20 to 50 years (mean age 31±10 years), were evaluated. Data collection consisted on the simultaneous capture of the swallowing audio with a microphone and of the swallowing videofluoroscopic image. The bursts of the swallowing acoustic signal were identified and their duration and the interval between them were later analyzed using a specific software, which allowed the simultaneous analyses between the acoustic wave and the videofluoroscopic image. Results: Three burst components were identified in most of the swallows evaluated. The first burst presented mean time of 87.3 milliseconds (ms) for water and 78.2 for the substance. The second burst presented mean time of 112.9 ms for water and 85.5 for the pasty substance. The mean interval between first and second burst was 82.1 ms for water and 95.3 ms for the pasty consistency, and between second and third burst was 339.8 ms for water and 322.0 ms for the pasty consistency. Conclusion: The software allowed the visualization of three bursts during the swallowing of healthy individuals, and showed that the swallowing signal in normal subjects is highly variable.

Electromyographic validation of the trapezius and serratus anterior muscles in military press exercises with middle grip

Electromyographic activity of the trapezius muscle and serratus anterior muscle was analysed in 24 male volunteers using a 2-channel TECA TE 4 electromyograp, during the execution of four different modalities of military press exercises with middle grip. The trapezius acted preferentially in the modalities standing press behind neck; and sitting forward and press behind neck, while SI did not show any significative difference among the modalities. The high levels of action potentials with which TS and SI acted justify the inclusion of these exercises in physical programmes.

Electromyographic analysis of the pectoralis major and deltoideus anterior in the inclined flying exercise with loads

Due to a shortage of textbooks with specific data on muscular activity concerning physical conditioning and sports, we analysed electromyographically the muscles pectoralis major and deltoideus anterior, bilaterally, in inclined flying exercises, during the concentric and eccentric phases, with external loads of 25, 50, 75 and 100% of the maximum load. The electromyographic analysis was performed in eleven male volunteers with MEDITRACE-200 surface electrodes connected to a six-channel biologic signal acquisition module coupled to a PC/AT computer. The electromyographic signals were processed and the obtained effective values were normalized through maximum voluntary isometric contraction. Statistically, the results showed that all the muscles studied presented significant differences between the concentric and the eccentric phases, with higher electromyographic activity during the concentric phase. By analysing the different loads for each muscle in both phases, significant electromyographic activity was observed for all muscles. When the effect of each load on each muscle during the concentric phase was analysed, it was noticed that the muscles on the left were more active than those on the right side, while in the eccentric phase the muscles had different behavior.

Mandible protrusion and decrease of TMJ sounds: An electrovibratographic examination

This study quantified by, electrovibratography, the amount of mandible protrusion required to decrease significantly temporomandibular joint (TMJ) vibratory energy as an aid in the diagnosis of the recapture of anteriorly displaced disk. Eighteen patients diagnosed as having anterior disk displacement with reduction and TMJ clicking were submitted to electrovibratographic examination at the first appointment and treated with a stabilizing appliance and anterior positioning appliance with 1 to 5 mm protrusion. Vibratory energy was checked in each of these positions. Baseline data were used as control. At the first appointment, the patients had vibrations with more elevated intensities at the middle and late phases of the mouth opening cycle. At only one clinical step, mandible protrusion was obtained with the anterior repositioning appliance, ranging from 1 to 5 mm protusion. At each new position, a new electrovibratographic exam was made. After the 5-mm mandibular projection, only 2 patients presented vibration, with means between 0.6 and 2.8 Hz. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). The outcomes of this study indicate that 3 mm is the minimum amount of mandible protrusion to significantly decrease the TMJ vibratory energy and to recapture the displaced articular disk.

Effects of discontinuous Percoll gradient centrifugation on the quality of bovine spermatozoa evaluated with computer-assisted semen analysis and fluorescent probes association

The objective of this study was to evaluate the quality of bovine frozen-thawed sperm cells after Percoll gradient centrifugation. Frozen semen doses were obtained from six bulls of different breeds, including three taurine and three Zebu animals. Four ejaculates per bull were evaluated before and after discontinuous Percoll gradient centrifugation. Sperm motility was assessed by computer-assisted semen analysis and the integrity of the plasma and acrosomal membranes, as well as mitochondrial function, were evaluated using a combination of fluorescent probes propidium iodide, fluorescein isothiocyanate-conjugated Pisum sativum agglutinin and 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide. The procedure of Percoll gradient centrifugation increased the percentage of total and progressive sperm motility, beat frequency, rectilinear motility, linearity and rapidly moving cells. In addition, the percentage of cells with intact plasma membrane and mitochondrial membrane potential was increased in post-centrifugation samples. However, the percentage of sperm cells with intact acrosomal membrane was markedly reduced. The method used selected the motile cells with intact plasma membrane and higher mitochondrial functionality in frozen-thawed bull semen, but processing, centrifugation and/or the Percoll medium caused damage to the acrosomal membrane.

Fluorescence Level of Composites Assessed by Computer Processing of Digital Images - ScanWhite

The human dentition is naturally translucent, opalescent and fluorescent. Differences between the level of fluorescence of tooth structure and restorative materials may result in distinct metameric properties and consequently perceptible disparate esthetic behavior, which impairs the esthetic result of the restorations, frustrating both patients and staff. In this study, we evaluated the level of fluorescence of different composites (Durafill in tones A2 (Du), Charisma in tones A2 (Ch), Venus in tone A2 (Ve), Opallis enamel and dentin in tones A2 (OPD and OPE), Point 4 in tones A2 (P4), Z100 in tones A2 ( Z1), Z250 in tones A2 (Z2), Te-Econom in tones A2 (TE), Tetric Ceram in tones A2 (TC), Tetric Ceram N in tones A1, A2, A4 (TN1, TN2, TN4), Four seasons enamel and dentin in tones A2 (and 4SD 4SE), Empress Direct enamel and dentin in tones A2 (EDE and EDD) and Brilliant in tones A2 (Br)). Cylindrical specimens were prepared, coded and photographed in a standardized manner with a Canon EOS digital camera (400 ISO, 2.8 aperture and 1/ 30 speed), in a dark environment under the action of UV light (25 W). The images were analyzed with the software ScanWhite©-DMC/Darwin systems. The results showed statistical differences between the groups (p < 0.05), and between these same groups and the average fluorescence of the dentition of young (18 to 25 years) and adults (40 to 45 years) taken as control. It can be concluded that: Composites Z100, Z250 (3M ESPE) and Point 4 (Kerr) do not match with the fluorescence of human dentition and the fluorescence of the materials was found to be affected by their own tone.

A current-mode based Field-Programmable Analog Array for signal processing applications

This paper presents a new approach to develop Field Programmable Analog Arrays (FPAAs),(1) which avoids excessive number of programming elements in the signal path, thus enhancing the performance. The paper also introduces a novel FPAA architecture, devoid of the conventional switching and connection modules. The proposed FPAA is based on simple current mode sub-circuits. An uncompounded methodology has been employed for the programming of the Configurable Analog Cell (CAC). Current mode approach has enabled the operation of the FPAA presented here, over almost three decades of frequency range. We have demonstrated the feasibility of the FPAA by implementing some signal processing functions.

Effect of refrigeration systems upon frozen bull sperm viability assessed by computer-assisted sperm analysis and fluorescent probes

Sperm cryopreservation success depends upon the maintenance of spermatozoa fertility potential. Sperm cells must preserve both integrity and functionality of several cell structures. The stabilization phase must allow the exit of water from the sperm cells via osmosis. This study aimed to compare the effect of refrigeration in the commercial refrigerator (CR) and the transport/refrigeration box (TRB) upon the viability of frozen bull sperm diluted in three different extenders (A, B and C). Ten Nellore bulls, Bos taurus indicus maintained in Artificial Insemination Center were used and the spermatozoa samples was assessed for Plasma Membrane Integrity and CASA evaluation. The stabilization phase (5 degrees C/4 hours) was performed in the CR as well as in the TRB, and then samples were exposed to nitrogen vapor during 20 minutes and then plunged into nitrogen. The statistical analysis was done using the variance analysis and the significance level was set at 5%. In the CR the post-thawing parameters for PM and ALH were higher (p < 0.05) in the extender A (glicine egg-yolk) and extender B (glicine egg-free) when compared with extender C (TRIS egg-yolk). As for BCF, STR and LIN, the parameters were higher (p < 0.05) in extender B than in C. Samples that were stabilized in the TRB presented higher post-thawing parameters (p < 0.05) for PM and LIN in extender A and extender B when compared with C. BCF and STR parameters were higher (p < 0.05) in extemder B when compared with C. Extender B samples had higher (p < 0.05) PMI when stabilized in CR. The findings in this experiment enable us to say that both CR and TRB were effective in keeping the viability of post-thawing bull semen.

Pattern recognition theory in nonlinear signal processing

A body of research has developed within the context of nonlinear signal and image processing that deals with the automatic, statistical design of digital window-based filters. Based on pairs of ideal and observed signals, a filter is designed in an effort to minimize the error between the ideal and filtered signals. The goodness of an optimal filter depends on the relation between the ideal and observed signals, but the goodness of a designed filter also depends on the amount of sample data from which it is designed. In order to lessen the design cost, a filter is often chosen from a given class of filters, thereby constraining the optimization and increasing the error of the optimal filter. To a great extent, the problem of filter design concerns striking the correct balance between the degree of constraint and the design cost. From a different perspective and in a different context, the problem of constraint versus sample size has been a major focus of study within the theory of pattern recognition. This paper discusses the design problem for nonlinear signal processing, shows how the issue naturally transitions into pattern recognition, and then provides a review of salient related pattern-recognition theory. In particular, it discusses classification rules, constrained classification, the Vapnik-Chervonenkis theory, and implications of that theory for morphological classifiers and neural networks. The paper closes by discussing some design approaches developed for nonlinear signal processing, and how the nature of these naturally lead to a decomposition of the error of a designed filter into a sum of the following components: the Bayes error of the unconstrained optimal filter, the cost of constraint, the cost of reducing complexity by compressing the original signal distribution, the design cost, and the contribution of prior knowledge to a decrease in the error. The main purpose of the paper is to present fundamental principles of pattern recognition theory within the framework of active research in nonlinear signal processing.

Computer-assisted analysis of cell proliferation markers in oral lesions

Abnormalities in any component of the cell cycle regulatory machine may result in oral. cancer, and markers of cell proliferation have been used to determine the prognosis of tumor progression. The aim of this study was to determine whether silver-stained nucleolar organizer region (AgNOR) and Ki-67 measurements could improve the assessment of growth rates in oral lesions. Eighty-three oral biopsies were studied, 20 of which were classified as fibrous inflammatory hyperplasia (FIH), 40 as leukoplakia (LKP) and 23 as oral. squamous cell carcinoma (OSCC). Within the LKP group, 22 out of 29 biopsies were diagnosed as non-dysplastic leukoplakia (LK) and 18 as dysplastic teukoptakia (DLK), presenting discrete, moderate and severe dysplasia. Ki-67 immunotabeting of the lesions increased steadily in the following order: FIH, DLK, LK and OSCC, indicating that Ki-67 is a good marker for predicting the protiferative fraction among benign, premalignant and malignant oral lesions. The median values of AgNOR parameters indicate that the morphometric index gives better results regarding the proliferative rate than the numerical one. A series of linear regressions between AgNOR parameters and Ki-67 showed positive associations. We conclude that a combination of Ki-67 and morphometric AgNOR analyses could be used as an aid in the determination of the protiferative status of oral epithelial. cells in oral cancer. (C) 2007 Elsevier GmbH. All rights reserved.

Galectin-3 Overexpression in Invasive Laryngeal Carcinoma, Assessed by Computer-assisted Analysis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)