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.

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.

Morphometric study by image analysis of Ag-stained nucleoli in thyroids bearing proliferating lesions

The Ag-NOR staining technique and image analysis were used to evaluate morphological parameters (area, perimeter and axis ratio) in nucleoli from normal thyroids and from thyroids bearing proliferating lesions (carcinomas, adenomas and hyperplasias). Regions with normal appearance located close to adenomatous and carcinomatous regions, in the thyroid of every patient, were also analyzed for comparison with the respective pathological regions and with normal thyroids. Statistical analysis of data for the nucleolar area and perimeter allowed the separation of adenomas and carcinomas from hyperplasias and normal tissue but not the two components in each of these two groups. However, if we look at the numbers, a sequence of increasing nucleolar mean areas in the order: normal, hyperplasia, adenoma and carcinoma may be observed, indicating the sequence of increasing rRNA requirements in these different kinds of cells. The axis ratio that denotes the nucleolar shape (round or oblong) did not show significant differences among tissues, suggesting that shape is not important in the characterization of these pathologies. Differences in nucleolar areas and perimeter between normal and affected regions from each patient were statistically significant for adenomas and carcinomas. When these normal regions were compared with the normal thyroids, significant differences were not obtained in the three evaluated parameters. The observations and their importance for histopathological diagnosis are discussed.

Evaluation epidermal p53 immunostaining by digital image analysis

Digital techniques have been developed and validated to assess semiquantitatively immunohistochemical nuclear staining. Currently visual classification is the standard for qualitative nuclear evaluation. Analysis of pixels that represents the immunohistochemical labeling can be more sensitive, reproducible and objective than visual grading. This study compared two semiquantitative techniques of digital image analysis with three techniques of visual analysis imaging to estimate the p53 nuclear immunostaining. Methods: Sixty-three sun-exposed forearm-skin biopsies were photographed and submitted to three visual analyses of images: the qualitative visual evaluation method (0 to 4 +), the percentage of labeled nuclei and HSCORE. Digital image analysis was performed using ImageJ 1.45p; the density of nuclei was scored per ephitelial area (DensNU) and the pixel density was established in marked suprabasal epithelium (DensPSB). Results: Statistical significance was found in: the agreement and correlation among the visual estimates of evaluators, correlation among the median visual score of the evaluators, the HSCORE and the percentage of marked nuclei with the DensNU and DensPSB estimates. DensNU was strongly correlated to the percentage of p53-marked nuclei in the epidermis, and DensPSB with the HSCORE. Conclusion: The parameters presented herein can be applied in routine analysis of immunohistochemical nuclear staining of epidermis. © 2012 John Wiley & Sons A/S.

Effect of digitization parameters on periapical radiographic image quality with regard to anatomic landmarks

The aim of this study was to evaluate the influence of digitization parameters on periapical radiographic image quality, with regard to anatomic landmarks. Digitized images (n = 160) were obtained using a flatbed scanner with resolutions of 300, 600 and 2400 dpi. The radiographs of 2400 dpi were decreased to 300 and 600 dpi before storage. Digitizations were performed with and without black masking using 8-bit and 16-bit grayscale and saved in TIFF format. Four anatomic landmarks were classified by two observers (very good, good, moderate, regular, poor), in two random sessions. Intraobserver and interobserver agreements were evaluated by Kappa statistics. Inter and intraobserver agreements ranged according to the anatomic landmarks and resolution used. The results obtained demonstrated that the cement enamel junction was the anatomic landmark that presented the poorest concordance. The use of black masking provided better results in the digitized image. The use of a mask to cover radiographs during digitization is necessary. Therefore, the concordance ranged from regular to moderate for the intraobserver evaluation and concordance ranged from regular to poor for interobserver evaluation.

Avaliação de parâmetros métricos oculares pela fotografia digital da face: uso do diâmetro da íris como unidade de referência

Purpose: To evaluate reproducibility and precision of ocular measurements by digital photograph analysis, in addition to the transformation of the measures according to the individual iris diameter as an oculometric reference. Methods: Twenty-four eyes have been digitally photographed in a standardized way at two distances. Two researchers have analyzed these printed images using a caliper and these digital forms by ImageJ 1.37 (TM). Several external ocular parameters were estimated (mm and as iris diameter) and methods of measurement compared regarding their precision, agreement and correlation. Results: Caliper and digital analysis of oculometric measures provided significant agreement and correlation, nevertheless the precision of digital measures was higher. The estimates of numeric transformation from oculometric measures according to individual iris diameter resulted in great correlation to caliper measures and high agreement when compared to different distances of taking the photographs. Conclusions: Facial digital photographs allowed oculometric precise and reproducible estimates, endorsing clinical research usefulness. Using iris diameter as individual oculometric reference disclosed high reproducibility when facial photographs were taken at different distances.

Úlceras crônicas dos membros inferiores: avaliação pela fotografia digital

OBJETIVO: Avaliar o desempenho da análise de imagem digital na estimativa da área acometida pelas úlceras crônicas dos membros inferiores. MÉTODOS: Estudo prospectivo em que foram mensuradas úlceras empregando o método planimétrico clássico, utilizando desenho dos seus contornos em filme plástico transparente, medida sua área posteriormente por folha milimetrada. Esses valores foram utilizados como padrão para a comparação com a estimativa de área pelas fotografias digitais padronizadas das úlceras e dos desenhos das mesmas em filme plástico. Para criar um referencial de conversão dos pixels em milímetros, foi empregado um adesivo com tamanho conhecido, adjacente à úlcera. RESULTADOS: foram avaliadas 42 lesões em 20 pacientes portadores de úlceras crônicas de membros inferiores. As áreas das úlceras variaram de 0,24 a 101,65cm². Observou-se forte correlação entre as medidas planimétricas e as fotos das úlceras (R²=0,86 p<0,01), porém a correlação das medidas planimétricas com as fotos digitais dos desenhos das úlceras foi ainda maior (R²=0,99 p<0,01). CONCLUSÃO: A fotografia digital padronizada revelou-se método rápido, preciso e não-invasivo capaz de estimar a área afetada por úlceras. A avaliação das medidas fotográficas dos contornos das úlceras deve ser preferida em relação à análise de sua fotografia direta.

Avaliação da posição do supercílio por medidas angulares

OBJETIVO: Avaliar a posição do supercílio em diferentes idades, utilizando medidas angulares. MÉTODOS: Foram avaliados indivíduos com idade de 4 a 6 anos (Grupo de crianças) e igual ou superior a 50 anos (Grupo de idosos), separados em faixas etárias, avaliando-se a posição do supercílio por meio de imagens digitais, utilizando medidas angulares. As imagens foram tomadas em posição primária do olhar, utilizando filmadora Sony Lithium, e posteriormente transferidas para computador MacIntosh G4 e processadas pelo programa NIH 1,58. Os parâmetros analisados foram: ângulo interno, externo e vertical da cauda do supercílio. As comparações foram entre sexos, faixas etárias e lateralidade. Os resultados obtidos foram submetidos à análise estatística. RESULTADOS: A comparação das medidas angulares mostrou que houve diferença significativa na posição da cauda do supercílio entre os grupos estudados quando comparados dentro do grupo com faixa etária semelhante. Porém, comparando-se crianças e adultos, houve diferença em todos os tipos de ângulos estudados. CONCLUSÕES: A posição do supercílio avaliada por medidas angulares mostrou diferenças entre crianças e idosos, revelando associação positiva com a idade.

Dimensões palpebrais antes e após blefaroplastia superior: avaliação quantitativa

OBJETIVO: Quantificar, usando o sistema de imagem digital, medidas palpebrais antes e após a cirurgia de blefaroplastia superior. MÉTODOS: Foram avaliadas 18 pálpebras de 9 pacientes atendidas no HC da FMB - UNESP, com idade entre 40 a 75 anos, do sexo feminino, portadoras de dermatocálase. Foram obtidas fotografias das pacientes antes e após 60 dias da blefaroplastia da pálpebra superior. As imagens foram transferidas para um computador e analisadas pelo programa Scion Image Frame Grabber. Os parâmetros avaliados foram: a altura da fenda palpebral em posição primária do olhar, altura do sulco palpebral superior e o ângulo palpebral lateral antes e depois de 60 dias da realização da cirurgia de blefaroplastia superior. RESULTADOS: Após a cirurgia, houve aumento da altura da fenda palpebral e do sulco palpebral superior. Contudo, o ângulo palpebral lateral não se alterou. CONCLUSÃO: A posição palpebral se altera após a blefaroplastia e o processamento de imagens digitais possibilita quantificar estas alterações, mensurando os resultados obtidos com a cirurgia.

Avaliação quantitativa das dimensões óculo-palpebrais em portadores de cavidade anoftálmica em uso de prótese ocular externa

OBJETIVOS: Avaliar o posicionamento palpebral em portadores de cavidade anoftálmica com e sem prótese ocular externa, utilizando o processamento de imagem digital. MÉTODOS: Dezoito pacientes foram avaliados qualitativa e quantitativamente na Faculdade de Medicina de Botucatu - Universidade Estadual Paulista - UNESP, com e sem a prótese externa. Usando imagens obtidas por filmadora e processadas usando o programa Scion Image, mediu-se a altura do sulco palpebral superior, a altura da fenda palpebral e os ângulos palpebrais dos cantos interno e externo. RESULTADOS: Pseudo-estrabismo e sulco palpebral superior profundo foram as alterações mais freqüentes ao exame externo. Houve diferença significativa em todas as variáveis estudadas, com diminuição da altura do sulco palpebral superior, aumento da área da fenda palpebral e aumento dos ângulos palpebrais interno e externo quando o paciente estava usando a prótese externa. CONCLUSÃO: Todos os pacientes avaliados apresentaram algum tipo de anormalidade órbito-palpebral, o que reflete a dificuldade em se proporcionar ao portador de cavidade anoftálmica um aspecto idêntico ao que existe na órbita normal. O processamento de imagens digitais permitiu avaliação objetiva das dimensões óculo-palpebrais, o que poderá contribuir nas avaliações seqüenciais dos portadores de cavidade anoftálmica.

CIE L*a*b*: comparison of digital images obtained photographically by manual and automatic modes

The aim of this study was to analyze the color alterations performed by the CIE L*a*b* system in the digital imaging of shade guide tabs, which were obtained photographically according to the automatic and manual modes. This study also sought to examine the observers' agreement in quantifying the coordinates. Four Vita Lumin Vaccum shade guide tabs were used: A3.5, B1, B3 and C4. An EOS Canon digital camera was used to record the digital images of the shade tabs, and the images were processed using Adobe Photoshop software. A total of 80 observations (five replicates of each shade according to two observers in two modes, specifically, automatic and manual) were obtained, leading to color values of L*, a* and b*. The color difference (AE) between the modes was calculated and classified as either clinically acceptable or unacceptable. The results indicated that there was agreement between the two observers in obtaining the L*, a* and b* values related to all guides. However, the B1, B3, and C4 shade tabs had AE values classified as clinically acceptable (Delta E = 0.44, Delta E = 2.04 and Delta E = 2.69, respectively). The A3.5 shade tab had a AE value classified as clinically unacceptable (Delta E = 4.17), as it presented higher values for luminosity in the automatic mode (L* = 54.0) than in the manual mode (L* = 50.6). It was concluded that the B1, B3 and C4 shade tabs can be used at any of the modes in digital camera (manual or automatic), which was a different finding from that observed for the A3.5 shade tab.

Three-dimensional reconstruction of fracture surfaces: Area matching algorithms for automatic parallax measurements

This paper describes two solutions for systematic measurement of surface elevation that can be used for both profile and surface reconstructions for quantitative fractography case studies. The first one is developed under Khoros graphical interface environment. It consists of an adaption of the almost classical area matching algorithm, that is based on cross-correlation operations, to the well-known method of parallax measurements from stereo pairs. A normalization function was created to avoid false cross-correlation peaks, driving to the true window best matching solution at each region analyzed on both stereo projections. Some limitations to the use of scanning electron microscopy and the types of surface patterns are also discussed. The second algorithm is based on a spatial correlation function. This solution is implemented under the NIH Image macro programming, combining a good representation for low contrast regions and many improvements on overall user interface and performance. Its advantages and limitations are also presented.

Inside-out vs. standard vein graft to repair a sensory nerve in rats

Nerve regeneration in a sensory nerve was obtained by the application of different techniques: inside-out vein graft (IOVG group) and standard vein graft (SVG group). These techniques provide a good microenvironment for axon regeneration in motor nerves, but their efficiency for regeneration of sensory nerves is controversial. The saphenous nerve was sectioned and repaired by the inside-out and standard vein graft techniques in rats. After 4, 12, and 20 weeks the graft and the distal stump were observed under electron microscopy. In each studied period, the pattern, diameters, and thickness of the myelin sheaths of the regenerated axons were measured in the graft and distal stump. A comparative study about the regenerated nerve fibers by these two different techniques was performed. Regenerated nerve fibers were prominent in both vein grafts 4 weeks after the surgical procedures. On the other hand, in the distal stump, regenerated nerve fibers were observed only from 12 weeks. In both inside-out vein graft and standard vein graft statistical difference was not observed about the diameters and thickness of the myelinated fibers after 20 weeks. On the other hand, the inside-out group had greater regenerated axon number when compared to the standard group. There is a capillary invasion in both graft and distal stump, especially in the IOVG group. The regenerated axons follow these capillaries all the time like satellite microfascicles. After 20 weeks, the diameters of regenerated fibers repaired by the standard vein graft technique were closer to the normal fibers compared to the inside-out vein graft. On the other hand, the pattern of these regenerated axons was better in the IOVG group.

A comparison of molar root canal preparation using stainless-steel and nickel-titanium instruments

Fifty-four extracted human mandibular molars were embedded and sectioned at two levels. The reassembled mesial root canals were prepared with stainless-steel hand K-files (Flexofiles) and either Nitiflex or Mity nickel-titanium hand K-files using a push-pull anticurvature filing technique. Each of the three experimental groups contained 36 mesial canals randomly distributed. Superimposed pre- and postinstrumentation cross-sectional root images were magnified using a stereomicroscope and transferred to a computer for measurement and statistical analysis. The direction and extent of canal center movement were evaluated. At the apical level, the groups produced no significant difference of direction of canal center movement. In cervical sections, all groups tended to move in a distolingual direction. The three groups, however, produced no significant difference in the cervical sections in the extent of canal center movement. In apical sections, Nitiflex produced the least canal center movement. Copyright © 1999 by The American Association of Endodontists.

Morphology of the ventral lobe of the prostate and seminal vesicles in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of prostate and seminal vesicles, which are the subject of this analysis in UCh ethanol-drinking rats. The prostate and seminal vesicles of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of the weight of the prostate and seminal vesicle, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of prostate and seminal vesicles and hypothalamus-pituitary axis of UCh rats.