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.     

Use of quantitative analysis as a method for differentiation between canine cutaneous apocrine adenomas and apocrine carcinomas on cytological smears

Eight canine cutaneous adenomas and eight canine cutaneous carcinomas were analysed by computer-assisted nuclear morphometry in Hemacolor-stained cytological specimens. 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 results indicated an increase in the mean values of investigated parameters from canine cutaneous apocrine adenomas (MNA, 75.65+/-2.22; MNP, 31.05+/-0.55; MND, 9.62+/-0.14; NR, 1.10+/-0.009) to canine cutaneous apocrine carcinomas (MNA, 88.78+/-11.29; MNP, 34.38+/-2.43; MND, 10.43+/-0.76; NR, 1.21+/-0.07). The statistical analysis revealed statistically significant differences between benign and malignant neoplastic cells (P<0.01). The statistical differences between investigated parameters (P<0.01) were also found between the metastasizing apocrine carcinomas and all other examined carcinomas. The results indicated that the computerized morphometry could be used as an effective auxiliary tool for differential diagnosis between canine cutaneous adenomas and carcinomas on cytological smears.     

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.     

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.     

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.     

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.     

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.     

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.     

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     

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.     

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.     

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.     

Fine needle aspiration cytology of pilomatricoma in three dogs

Background — Fine needle aspiration cytology is being used for the diagnosis of various neoplasms, but we are unaware of reports dealing with the cytologic features of canine pilomatricoma.
Objective — The purpose of this report was to describe the cytologic features of pilomatricoma in 3 dogs.
Methods — Fine-needle aspirates were obtained from cutaneous masses using a 25-ga needle. Smears were prepared and stained with May-Grünwald-Giemsa and hematoxylin and eosin. The cutaneous masses were excised and routinely processed for histologic examination.
Results — Cytologic features of all 3 tumors included high cellularity and numerous clusters of tightly arranged of basaloid cells with evenly distributed chromatin and small distinct nucleoli surrounding sheets of "ghost cells". Ghost cells were characterized by a central unstained zone corresponding to the site previously occupied by the nucleus. Amorphous keratinized material was observed in 1 tumor. Histopathologic findings confirmed the diagnosis of pilomatricoma in all 3 cases.
Conclusions — Basaloid cells in association with ghost cells are important cytopathologic criteria that may provide a definitive cytologic diagnosis of pilomatricoma in dogs, and may help avoid a false diagnosis of malignancy.     

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.     

Assessment of spermatozoa in fertile alpaca (Vicugna pacos) males: Study of sperm head morphometry using a nonautomated digital method and sperm morphology based on strict criteria

Although computer‐assisted systems for sperm morphometry and morphological analysis are important tools in the study of male fertility, their use in extensive systems in alpacas is limited by factors such as the expense of equipment and the high altitudes of the Andean region. The objectives of this study were to evaluate alpaca sperm head morphometry using a nonautomated digital method and determine the frequency of sperm abnormalities based on strict criteria for sperm morphology in fertile male alpacas. Ejaculates (n = 15) from seven alpacas were collected, and sperm smears stained with modified Papanicolaou were processed. For morphometric analysis, 3,000 sperm (200 cells/sample) images were captured at 400× magnification and Quick Photo MICRO 3.0 software was used for manual measurement of basic (sperm head length, width, perimeter and area) and derived variables (ellipticity, shape factor, elongation and regularity). For morphology assessment, smears were observed at 1000× magnification according to WHO and strict criteria. Average morphometric parameters were length 5.48 μm, width 2.99 μm, perimeter 13.62 μm, area 12.43 μm², ellipticity 1.86, shape factor 1.20, elongation 0.29 and regularity 1.05. Significant between‐individual and within‐individual differences were found in morphometric parameters. Based on morphometric study, sperm heads were classified as elliptical or normal (49%), long (18%), short (2%), pyriform (12%), round (9%), large (6%) and small (4%). Morphological analysis found no additional sperm head defects in 49% of normal sperm obtained by morphometry, although a 4% incidence of neck/mid‐piece defects and a 16% incidence of principal‐piece defects were found. We conclude that sperm head morphometry assessment in fertile alpacas using a nonautomated digital method is feasible, and that defects in sperm heads constitute the main morphological alteration (>50% of the sperm population), based on WHO and strict criteria.     

Ejaculate Fractioning Effect on Llama Sperm Head Morphometry as Assessed by the ISAS® CASA system

South American camelid sperm characteristics are poorly known compared with those of other domestic animals. The long‐term duration of ejaculation makes difficult to gather all the seminal fluid, implying possible ejaculation portion losses. Thus, the aim of this research was to evaluate the characteristics of the morphology and morphometry of the spermatozoa change during ejaculation. The morphometric characterization was tested on nine specimens of the Lanuda breed, using a special artificial vagina. In five of the animals, a fractioning of the ejaculate was performed by taking samples every 5 min. for a total of 20 min. Air‐dried seminal smears were stained with Hemacolor and mounted permanently with Eukitt. Morphometric analysis was carried out with the morphometry module of the ISAS^® CASA system. Almost 350 cells were analysed per sample, with a total number of 3207 spermatozoa. Mean values were given as follows: length: 5.51 μm; width: 3.38 μm; area: 17.75 μm²; perimeter: 14.8 μm; ellipticity: 0.24; elongation: 0.56; rugosity: 0.87; regularity: 1.07; and shape factor: 1.41. Different animals showed differences in their morphometric values. When we compared the values from different fractions, only two samples showed differences in morphometric parameter values and four samples showed differences in shape parameters. Multivariate analysis allowed the size classification of the cells into three classes and five classes of shapes. The distribution of classes among fractions showed no differences. Despite the individual morphometric differences observed in some fractions, the characteristics of the sperm head morphometry can be considered constant along the ejaculatory period in the llama.     

The Effect of Triple Pelvic Osteotomy on the Articular Contact Area of the Hip Joint in Dysplastic Dogs: An in Vitro Experimental Study

Objective —To investigate the effect of triple pelvic osteotomy (TPO) on articular contact area and acetabular coverage of dysplastic hip joints in dogs.
Study Design —Articular contact area and femoral head coverage by the acetabulum were computed in vitro in normal and dysplastic canine hips. The effect of TPO on articular contact and coverage was then analyzed in the dysplastic hips.
Sample Population—Five normal and six dysplastic canine cadaver specimens.
Methods —Contact area and coverage of loaded hips were computed using serial computed tomography scan images before and after TPO. Three angles of acetabular ventroversion (AVV) were studied (20°, 30°, and 40°). Using a custom-designed hinge plate, angles of spontaneous hip reduction in dysplastic hips were compared with previously recorded angles of reduction determined by the Ortolani test.
Results —Contact area significantly increased from 0° to 30° of AVV, then remained virtually unchanged. Coverage significantly increased from 0° to 20° of AVV. Both contact and coverage of normal hips were similar, yet significantly smaller than those of dysplastic hips once reduction had occurred. The experimental angles of reduction were significantly smaller and poorly correlated with the angles of reduction determined by the Ortolani test. Although coverage continued to increase with AVV, the actual joint contact area did not significantly vary after relocation of the femoral head.
Conclusions —This study suggests that increasing AVV beyond 20° does not significantly improve the beneficial effects of TPO and therefore should be carefully weighed against increased risks of postoperative complications associated with large angles of AVV.