Quantitative analysis in spontaneous canine anal sac gland adenomas and carcinomas

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas. The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.     

Quantitative analysis in spontaneous canine anal sac gland adenomas and carcinomas

Stained cytological specimens from 7 canine anal sac gland adenomas and 11 canine anal sac gland carcinomas were analyzed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine anal sac gland adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine anal sac gland adenocarcinomas.The results indicated that (1) MNA, MNP, MND and NR could be used as effective auxiliary tools for differential diagnosis between canine anal sac gland adenomas and adenocarcinomas, and (2) MNA, MNP and MND are reliable prognostic indicators for canine anal sac gland adenocarcinomas.     

Nuclear cytomorphometry in feline mammary gland epithelial tumours

Stained cytological specimens from 35 feline mammary gland epithelial tumours (4 adenomas, 11 tubulopapillary carcinomas 13 solid carcinomas and 7 cribriform carcinomas) were analysed by computer-assisted nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear diameter (MND) and nuclear roundness (NR) were calculated. The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between benign and malignant feline mammary gland epithelial tumours, and (2) the prognostic value of nuclear morphometry in feline mammary carcinomas. The results indicated that MNA, MNP, MND and NR could be a useful adjunct in diagnosis but are not reliable prognostic indicators for feline mammary gland carcinomas.     

Nuclear morphometry in cytological specimens of canine ceruminous adenomas and carcinomas

Stained cytological specimens from eight canine ceruminous adenomas and eight canine ceruminous carcinomas were analysed by computer‐assisted nuclear morphometry. Three carcinomas had metastases in regional lymph nodes at the time of the diagnosis. The morphometric parameters evaluated in this study were mean nuclear area (MNA, µm²), mean nuclear perimeter (MNP, µm), mean nuclear diameter (D mean, µm), minimum nuclear diameter (D min, µm) and maximum nuclear diameter (D max, µm). The study aimed to evaluate (1) the possibility of using nuclear cytomorphometry as an auxiliary diagnostic method to differentiate between canine ceruminous adenomas and adenocarcinomas, and (2) the prognostic value of nuclear morphometry in canine ceruminous carcinomas. The results indicated that (1) MNA, MNP, D mean, D min and D max could be used as effective auxiliary tools for differential diagnosis between canine ceruminous adenomas and adenocarcinomas, and (2) MNA, MNP, D mean and D max are reliable prognostic indicators for canine ceruminous adenocarcinomas.     

Comparative morphometric analysis of recurrent and nonrecurrent canine basal cell carcinomas: a preliminary report

Background: Most reports of canine basal cell carcinomas (BCCs) focus on morphologic appearance rather than biologic behavior. Rare recurrences and no metastases have been reported in dogs. Quantitative nuclear morphometry may be useful in predicting tumor recurrence. Objective: The aim of the present study was to compare quantitative nuclear parameters of canine BCCs that did not recur within 60 weeks of excision with those that recurred. Methods: Cytologic specimens obtained from spontaneous BCCs from 11 dogs were analyzed by computerized nuclear morphometry. The dogs were monitored by their owners over a period of 60 weeks to detect local recurrence of the tumor; recurrent tumors were confirmed histologically. For each initial tumor specimen, nuclei of at least 100 neoplastic cells were measured by 2 independent observers, and mean nuclear area (MNA), mean nuclear perimeter (MNP), and mean nuclear diameter (MND) were calculated. Results: Six nonrecurrent and 5 recurrent tumors were analyzed. Neoplastic cells from BCCs that subsequently recurred had higher MNA (102.41 ± 4.57 μm²), MNP (36.27 ± 0.61 μm), and MND (11.21 ± 0.27 μm) than cells from nonrecurrent BCCs (MNA 87.66 ± 4.79 μm², MNP 33.51 ± 0.78 μm, MND 10.36 ± 0.29 μm) (P<.001; Mann–Whitney U‐test). Conclusion: Based on these preliminary results, nuclear morphometry may be a useful tool to predict local recurrence of BCCs in dogs.     

Nucleomorphometric analysis of feline basal cell carcinomas

Twenty-four feline spontaneous basal cell carcinomas (BCCs) were analyzed by computerized nuclear morphometry. The study included 15 non-recurrent and 9 recurrent tumours. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP) and mean nuclear diameter (MND) were calculated. The analysis of data of the non-recurrent BCCs and the recurrent tumours revealed statistically significant differences between those groups (p<0.001) as well as between infiltrative and clear types of BCCs (p<0.05). The results indicate that nuclear morphometry is able to predict recurrent tumour growth and helps to differentiate histological subtypes of BCCs in cats.     

Comparison of nuclear morphometric parameters in cytologic smears and histologic sections of spontaneous canine tumors

Background — Nuclear morphometry may provide useful diagnostic and prognostic information for neoplasms in animals. Most available data have been obtained from histologic sections. Nuclear morphometry of cytologic smears may provide important pre-operative information.
Objectives — The goal of this study was to compare nuclear morphometric parameters in cytologic smears and histologic sections from spontaneous canine tumors.
Methods — Mean nuclear area (MNA), mean nuclear perimeter (MNP), mean nuclear form factor (FF; nuclear perimeter²/4π nuclear area) and their respective SDs were assessed by image analysis of both hematoxylin and eosin-stained histologic sections and May-Grünwald-Giemsa-stained cytologic smears from the same case in 20 spontaneous canine tumors of different histogenesis. The above parameters were selected as being the best morphometric tools for measuring variation in shape and size in cells after neoplastic transformation. Data were compared by ANOVA with P<.01 considered significant.
Results — There was a significant difference between histologic and cytologic specimens for MNA, MNP, and their SDs. Only the differences between FF and the SD of FF were not statistically significant.
Conclusions — Only nuclear morphometric data related to nuclear shape and nuclear shape variability are comparable between histologic and cytologic specimens. Nuclear area and perimeter may be affected by the different fixation and smear preparation techniques used in histology and cytology.     

Morphometrical approach for predicting regional lymph node micrometastatic load in canine mast cell tumours: preliminary results

Canine cutaneous mast cell tumours (MCTs) have a variable biologic behaviour, and accurate staging is necessary to dictate therapy and predict outcome. Regional lymph node (RLN) involvement is a relevant prognostic factor. While obvious lymph node (LN) metastases are relatively easy to be diagnosed, micrometastatic disease recognition is challenging. The main aim of the study was to evaluate the number of mast cells (MCs) in the LNs of clinically healthy dogs ( n = 4, group 1), dogs with inflammatory diseases ( n = 31, group 2) and dogs with cutaneous MCT ( n = 27, group 3), including animals with no RLN metastases (subgroup 3.1), those with occasional MCs in RLNs (3.2) and those with obvious RLN metastasis (3.3). MCs also were morphometrically evaluated for the following nuclear parameters: mean nuclear area (MNA), mean nuclear perimeter (MNP), largest to smallest diameter length (LS ratio), mean nuclear form factor and coefficient of variation of nuclear area. The average percentages of MCs were 0.0 and 0.01 in groups 1 and 2, respectively, and 0.07, 2.4 and 47.1 in subgroup 3.1, 3.2 and 3.3. MNA and MNP were significantly higher in subgroup 3.3 than in group 2 ( P < 0.05). MNA and MNP in subgroup 3.2 suggested the presence of neoplastic MCs; this prediction of micrometastatic load correlated with outcome. Analysis of preliminary results shows that nuclear morphometry is useful to detect micrometastatic disease in RLN of dogs bearing cutaneous MCTs.     

Computerized morphometry of mean nuclear diameter and nuclear roundness in canine mammary gland tumors on cytologic smears

BACKGROUND: The use of computer-based image analysis systems in veterinary oncology has increased. Computerized morphometry is a part of image analysis that describes geometric figures of cellular structures in any dimension. Most investigators have performed morphometric analysis on histologic specimens. Computer-assisted nuclear cytomorphometry can provide important preoperative information on neoplastic lesions in animals. OBJECTIVES: The aim of this study was to define whether the morphometric parameters of mean nuclear diameter and nuclear roundness could be used to differentiate benign from malignant canine mammary gland tumors on cytologic specimens. METHODS: Mean nuclear diameter and nuclear roundness were determined by computer-assisted morphometry of epithelial cells in Hemacolor-stained cytologic smears from normal canine mammary gland (n = 7) and from canine mammary adenomas (n = 8), tubulopapillary carcinomas (n = 9), and solid carcinomas (n = 6). Data were analyzed by the Mann-Whitney U test. RESULTS: Significant differences (P <.001) were found in mean nuclear diameter and nuclear roundness among all tumor types and in comparison with normal canine mammary gland epithelial cells (except for nuclear roundness between tubulopapillary carcinomas and solid carcinomas). CONCLUSIONS: The morphometric parameters of mean nuclear diameter and nuclear roundness can be used in the preoperative differentiation of benign from malignant canine mammary gland tumors.     

Computerized Cytomorphometric Analysis of Nuclear Area,Nuclear Perimeter and Mean Nuclear Diameter in Spontaneous Canine Mammary Gland Tumours

Fifty-two spontaneous canine mammary gland tumours (fibroadenomas (n = 8), tubulopapillary carcinomas (n = 9), solid carcinomas (n = 6), anaplastic carcinomas (n = 7), fibrosarcomas (n = 9), liposarcomas (n = 9) and osteosarcomas (n = 4) were analysed by computer-assisted nuclear morphometry in Hemacolor-stained cytological specimens. Computerized cytomorphometry was performed and the nuclear area, nuclear perimeter and mean nuclear diameter of investigated tumours were assessed. A minimum of 100 nuclei per lesion were examined. The statistical analysis revealed statistically significant differences between benign and malignant neoplasms. The results indicated that computer-assisted nuclear morphometry could be used as an additional method for differentiation of benign from malignant canine mammary gland tumours in cytological specimens.     

Quantitative morphology in canine cutaneous soft tissue sarcomas

Stained cytological specimens from 24 dogs with spontaneous soft tissue sarcomas [fibrosarcoma (n = 8), liposarcoma (n = 8) and haemangiopericytoma (n = 8)], and 24 dogs with reactive connective tissue lesions [granulation tissue (n = 12) and dermal fibrosis (n = 12)] were analysed by computer‐assisted nuclear morphometry. The studied morphometric parameters were: mean nuclear area (MNA; µm²), mean nuclear perimeter (MNP; µm), mean nuclear diameter (MND mean; µm), minimum nuclear diameter (D_min; µm) and maximum nuclear diameter (D_max; µm). The study aimed to evaluate (1) possibility for quantitative differentiation of soft tissue sarcomas from reactive connective tissue lesions and (2) by using cytomorphometry, to differentiate the various histopathological soft tissue sarcomas subtypes in dogs. The mean values of all nuclear cytomorphometric parameters (except for D_max) were statistically significantly higher in reactive connective tissue processes than in soft tissue sarcomas. At the same time, however, there were no considerable differences among the different sarcoma subtypes. The results demonstrated that the quantitative differentiation of reactive connective tissue processes from soft tissue sarcomas in dogs is possible, but the same was not true for the different canine soft tissue sarcoma subtypes. Further investigations on this topic are necessary for thorough explication of the role of quantitative morphology in the diagnostics of mesenchymal neoplasms and tumour‐like fibrous lesions in dogs     

A case of canine apocrine sweat gland adenoma, clear cell variant

A cutaneous mass at the base of the retroauricular region of a 4-year-old, female Golden Retriever was examined pathologically. Histologically, the mass formed multiple nodules consisting of a proliferation of large clear cells with abundant cytoplasm. Mitotic figures among the neoplastic cells were very sparse. The large clear cells were intensely positive for cytokeratins (AE1/AE4, cytokeratin 8 and 18) and moderately positive for lysozyme and contained periodic acid-Schiff-positive granules in the cytoplasm. In addition, small flat cells lined the islands of neoplastic large clear cells, and these were strongly positive for alpha-smooth muscle actin and vimentin, and some were positive for cytokeratin (AE1/AE4), suggesting they were myoepithelial cells. No local recurrence or metastasis has been recognized during the 18 months since surgical excision. On the basis of these findings, the present tumor was diagnosed as apocrine sweat gland adenoma, clear cell variant. There have been few previous reports of canine apocrine adenomas showing a clear cell morphology.     

Nuclear Morphometry in Relation to Lymph Node Status in Canine Mammary Carcinomas

The assessment of nuclear area and nuclear shape by morphometric analysis, has been investigated in 40 canine mammary carcinomas in relation to their metastatic behaviour to regional lymph-nodes. The tumours were reviewed by two experienced pathologists blinded regarding their lymph-node status, and were classified according to the histogenetically based criteria suggested by Benjamin et al. (1999). Twenty of these tumours showed lymph-node metastases (node-positive), and the other twenty were node-negative. Node-positive tumours included 6 simple adenocarcinomas, 10 ductular carcinomas, 2 anaplastic carcinomas and 2 carcinomas in mixed tumours; node-negative tumours included 18 adenocarcinomas %96, 10 simple adenocarcinomas, 8 complex adenocarcinomas %96, and 2 carcinomas in mixed tumours. Node-positive tumours showed MNA and mean SDA values significantly higher (p<0.001) than node-negative carcinomas. Data of this study, seems to confirm the importance of an histogenetically based classification in canine mammary tumours, also suggesting that morphometry may increase our prognostic performances allowing a reproducible method for detecting individual tumours with higher metastatic potential.     

Fractal dimension of canine mammary gland epithelial tumors on cytologic smears

BACKGROUND: Fractal geometry is a tool that can be used for describing, modeling, analyzing, and processing irregular and complex figures. Past investigations in medicine have revealed that fractal analysis could also be applied in tumor pathology to characterize irregular boundaries of the nuclei of tumor cells. OBJECTIVE: The aim of this study was to define whether the fractal dimension parameter could be used on cytologic specimens to differentiate benign from malignant canine mammary gland epithelial tumors. METHODS: The fractal dimension of nuclear surface was determined by computer-assisted morphometry on Hemacolor-stained cytologic smears obtained by fine needle aspiration of normal canine mammary gland epithelial cells, and cells in mammary adenomas, tubulopapillary carcinomas, solid carcinomas, and anaplastic carcinomas. Data were analyzed by Mann-Whitney U test. RESULTS: Significant differences (P <.001) were observed in mean fractal dimension among all tumor types and in comparison with normal canine mammary gland epithelial cells (except for the fractal dimension between solid carcinomas and anaplastic carcinomas). CONCLUSION: The morphometric parameter, fractal dimension, could help in the diagnostic discrimination between benign and malignant canine mammary gland epithelial tumors on cytologic specimens.     

Expression of oestrogen and progesterone receptors in canine sebaceous gland tumours

Sebaceous gland oestrogen α (ERα) and progesterone (PR) receptor expression was examined immunohistochemically in 26 and 32 dogs respectively with sebaceous gland hyperplasia/adenomas, epitheliomas and carcinomas, and in the glands of 10 healthy controls. The mean percentage of ERα positive nuclei in control sebaceous glands was 21.31% compared with 11.5% in hyperplasia/adenoma‐type lesions, although these values were not statistically different. In sebaceous gland epitheliomas and carcinomas, positive basal cells represented 7.86% and 3.53% of neoplastic cells respectively and these mean percentages were significantly lower in epitheliomas (P < 0.024) and carcinomas (P < 0.015) than in controls. The mean percentage of PR‐positive nuclei in control sebaceous glands was 23.96%, similar to the 22.07% found in hyperplasia/adenoma‐type lesions. In sebaceous gland epitheliomas and carcinomas, positive cells were scarce and represented 13.5% and 4.06% of neoplastic cells respectively. Differences in the percentage of positive cells between normal and pathological glands reached statistical significance for carcinomas (P < 0.043). In the control group there was greater PR (P < 0.001) and ERα expression (P < 0.014) in sebaceous glands in female dogs. The PR and ERα immunoreactivity in each category of neoplastic lesions could not be analysed because sample size was too small but when all the sebaceous gland tumours were grouped and analysed, no sex difference was found. The results suggest that oestrogen and progesterone receptor expression is reduced in some canine sebaceous gland tumours. These changes may represent a contributing factor for tumour growth or simply be a consequence of tumour progression.     

Indicators of malignancy of canine adrenocortical tumors: histopathology and proliferation index

Tumors of the adrenal cortex account for 10-20% of the naturally occurring Cushing's syndrome diagnosed in dogs. Differentiating between adrenocortical adenoma and carcinomas is often difficult. The purposes of this study were to determine which histopathologic criteria can be used as markers for malignancy in canine adrenocortical tumors and the relevance of the proliferation marker, Ki-67, for differentiation between cortical adenomas and carcinomas. Twenty-six adrenocortical carcinomas, 23 adenomas, and 11 normal adrenal glands were examined. Morphologic criteria significantly associated with adrenocortical carcinomas included a size larger than 2 cm in diameter, peripheral fibrosis, capsular invasion, trabecular growth pattern, hemorrhage, necrosis, and single-cell necrosis, whereas hematopoiesis, fibrin thombi, and cytoplasmic vacuolation were significantly associated with adrenocortical adenomas. The mean (+/- SD) proliferation index, measured by immunohistochemistry for the Ki-67 antigen, was 9.3 +/- 6.3% in carcinomas, 0.76 +/- 0.83% in adenomas, and 0.58 +/- 0.57% in normal adrenal glands. The Ki-67 proliferation index was significantly higher in carcinomas compared with adenomas and normal adrenal glands. A threshold value of the proliferation index of 2.4% reliably separated carcinomas from adenomas. Based on these results, it appears that thorough evaluation of morphologic features combined with immunohistochemical assessment of the proliferation index is extremely useful for differentiating between adrenocortical adenomas and carcinomas in dogs.     

Expression of p63 normal canine skin and primary cutaneous glandular carcinomas

p63, a recently identified homologue of the p53 protein, is expressed consistently in basal cells of several human multilayered epithelia. In this study, expression of p63 was determined in 31 primary cutaneous glandular carcinomas, including sebaceous, perianal (hepatoid) gland, apocrine and ceruminous carcinomas, as well as their adjacent normal skin. Similar to humans, p63 is a reliable marker for basal and myoepithelial cells in canine epidermis, cutaneous appendages and malignant apocrine and ceruminous gland neoplasms. In sebaceous carcinomas, not only basal cells, but also some sebocytes, showed nuclear staining for p63. Most mature epithelial cells in perianal gland carcinomas exhibited strong p63 expression. Based on these findings, basal/myoepithelial cells could be involved in the oncogenesis of these tumours and p63 might be used as a diagnostic marker in these lesions.     

Neoplasms of the apocrine sweat glands in 44 dogs and 10 cats

Neoplasms of the apocrine sweat glands accounted for 2.0 per cent and 3.6 per cent, respectively, of all canine and feline skin neoplasms diagnosed during a period of three years. They occurred in dogs from six to 17 years of age of both sexes, and golden retrievers appeared to be predisposed; they occurred in cats from six to 17 years of age and there were no breed or sex predilections. In both species, the neoplasms were usually solitary and occurred anywhere on the body; they were nearly always carcinomas and histologically were usually of the solid type. There were no clinical measurements that made it possible to distinguish reliably between benign and malignant lesions. No distant metastases were recorded, even though 22.5 per cent of the canine carcinomas had invaded the lymphatic system.     

Morphometry of canine cutaneous mast cell tumors

Twenty-four canine cutaneous nodules, diagnosed as mast cell tumors by fine-needle aspiration biopsy and confirmed by histopathologic analysis by staining with hematoxylin and eosin (HE) and toluidine blue, were analyzed by computerized nuclear morphometry on panoptic- and HE-stained cytopathology slides. Two hundred nuclei per lesion were examined. The morphometric parameters investigated were nuclear area, mean diameter, perimeter, regularity factor, and ellipticity factor. Lesions were graded as I (well differentiated), II (intermediate differentiation), or III (poorly differentiated) according to the following morphologic features: invasiveness, cellularity and cellular morphology, mitotic index, and stromal reaction. Nuclear morphometric results were then compared with histopathologic grades. Values of nuclear area, mean diameter, and perimeter increased with increase in histopathologic grade, but statistical analysis revealed significant differences only between grades II and III and between grades I and III when HE was used (P < 0.01) and between grades I and III with panoptic stain (P < 0.05). The ellipticity factor and regularity factor did not reveal significant differences between histopathologic grades. The results indicate that nuclear morphometric analysis, in combination with the rapid and inexpensive cytopathology technique, can help in mast cell tumor grading, thus contributing to the establishment of a more precise prognosis and treatment.     

Histopathological and immunohistochemical studies of apocrine sweat gland adenocarcinomas in cats

Seven adenocarcinomas of apocrine sweat glands among the 67cutaneous tumours of cats were evaluated between 1980 and 2005. Histopathologically, three tumours were solid, tubular, tubular‐ductal, tubular‐cystic and papillar‐cystic types were also diagnosed each one. Cuboidal and squamous epithelial cells surrounding the cystic lumina and their papillar extensions into the cyst lumens were evident in almost all tumour types. Besides, fibrovascular stroma which surrounding neoplastic foci showed proliferation and stromal invasion. Immunohistochemical features of these tumours were examined with pan‐cytokeratin, vimentin and carcinoembryonic antigen.