Quantification of morphology of canine circumanal gland tumors: a fractal based study

Abstract

Circumanal gland tumors are very common neoplasms of dogs. Their classification relies on microscopic examination and is further supported by a few immunohistochemical markers that help indicate their prognosis. However, new additional tests would be highly useful. The purpose of this study was to develop such a test using fractal analysis which is increasingly being applied in science, especially in the field of biomedicine. A total of 53 circumanal gland tumors were chosen from our department archives. After a precise histological classification according to the World Health Organization classification, the number of de novo classified samples was as follows: 15 adenomas, 11 epitheliomas, 21 well differentiated carcinomas, 6 poorly differentiated carcinomas. Ten samples of normal circumanal gland were also included as control. All samples were immunohistochemicaly stained with vimentin. All immunohistochemical reactions were photographed at two different magnifications -100X and 400X- and converted to 1 bit in black and white (bitmap) images, thus enhancing the positive vimentin reactions. These images were used for the assessment of fractal dimension applying the box counting method and computer software Fractalyse. To determine the significance of results, conventional statistics were performed using Statistica software.The overall vimentin stain score was significantly higher in epitheliomas and carcinomas than in normal circumanal glands (CG) or adenomas. Mean values of fractal dimension estimated at magnification 100X and 400X were as follows: normal CG 1.318 and 1.372, CG adenomas 1.384 and 1.408, CG epitheliomas 1.547 and 1.597, CG well differentiated carcinomas 1.569 and 1.607, CG poorly differentiated carcinomas 1.679 and 1.723. Significant differences (at level of 5%) of these values were observed between individual groups of CG adenomas or normal CG, and epitheliomas or carcinomas.The above results indicate vimentin immunohistochemistry staining and assessment of fractal dimension as an ancillary diagnostic method of choice when discerning between benign and malignant tumors of circumanal glands. Additional development of the method of fractal dimension assessment may yield a possibility for this tool to successfully discern between all of the types of CG tumors.

Is there still room for novelty, in histochemical papers?

“…The key role of the fixation/embedding procedures for appropriate specimen preparation was demonstrated in some studies, 99-104 and particular attention was paid to technical improvements in antigen preservation and retrieval for immunohistochemical applications, 105-110 and in multiple histochemical staining. 111-113 Imaging techniques were the subject of some articles. 114-116 Raman microspectroscopy was used to investigate stenotic aortic valve leaflets to get information on the composition and distribution of accumulated lipids, in the attempt to correlate their presence with mineralization as a precocious diagnostic marker.…”

Fractal Dimension in Liver Histological Findings of Wistar Rats Experimentally Intoxicated with Venom of Crotalus durissus terrificus

“…A new diagnostic tool called fractal analysis has been cited in some studies as a method of quantifying changes in cells such as retina [11], hepatocytes [27], renal and pulmonary vasculature [10,17,34], cardiac studies [12,18,26,33,35] and neoplasms [7].…”

>b<Avaliação da concordância interobservadores de critérios morfológicos no diagnóstico de tumores de glândula hepatoide em cães e determinação do valor prognóstico de aspectos clínico-patológicos e marcadores imuno-histoquímicos>/b<

“…Um caminho interessante a ser investigado é o mecanismo de transição epitélio-mesenquimal, já que há alguns indícios em literatura atual de que ele possa ocorrer com frequência em neoplasias de glândula hepatoide (ŠOŠTARIĆ-ZUCKERMANN et al, 2016), e a investigação deste processo por meio de marcadores imuno-histoquímicos, como vimentina, citoqueratinas e E-caderina, tem demonstrado valor prognóstico para neoplasias de outras origens (GAMA; ALVES; BONGIOVANNI et al, 2013;RISMANCHI et al, 2014).…”

“…A expressão de vimentina em células neoplásicas em tumores de glândula hepatoide tem sido estudada por outros autores, e o mecanismo de EMT tem sido proposto como provável justificativa para esta ocorrência, mas a concomitante perda de marcadores epiteliais não foi investigada (ŠOŠTARIĆ-ZUCKERMANN et al, 2016). No presente trabalho, foi aventada a hipótese de que uma maior expressão imuno-histoquímica de vimentina em tumores hepatoides estaria acompanhada de uma menor expressão de marcadores epiteliais, tais como pancitoqueratina e E-caderina.…”

“…A associação entre a perda da expressão destas proteínas e uma maior atividade proliferativa ainda precisa ser melhor investigada, já que resultados discrepantes tem sido relatados. demonstraram correlação negativa entre a expressão de E-caderina e β-catenina e contagem de mitoses, enquanto que Brunetti et al (2005) SARAIVA, 2011; PIEPER et al, ŠOŠTARIĆ-ZUCKERMANN et al, 2016).…”