Comparison of histology and clinical variables to DNA ploidy in canine mammary tumors

Flow cytometric DNA analysis was done on 132 canine mammary tumors from 99 dogs to evaluate the relation to histology and to clinical staging. Seventy-one tumors (54%) were histologically malignant; 38 (54%) of these were aneuploid and 33 (46%) were diploid. Fifty-two (39%) tumors were histologically benign, of which 45 (87%) were diploid and seven (13%) aneuploid. There were nine dysplastic mammae (7%); two were aneuploid and the rest diploid. DNA indices varied from 0.72 to 2.35. Of 58 mammary carcinomas, 25 (43%) were diploid and 33 (57%) were aneuploid (of the latter, 16 showed hypodiploidy and 17 hyperdiploidy with a predominance between DNA index 1.10 and 1.50). Three tumors (two carcinomas and one malignant mixed tumor) were multiploid with two aneuploid cell populations. The histological type varied within eight tumors, and in four of these the DNA index also varied. DNA indices varied within three tumors with uniform morphology. No correlation was found between DNA index and age of the dogs, nor between DNA index and tumor size. No significant differences were found between DNA index and histology, tumor growth pattern, or tumor location. Benign tumors were smaller than carcinomas, which were smaller than malignant mesenchymal tumors. Tumors growing adherent to the skin were larger than those not adherent to the skin. The regional lymph nodes were examined in 33 cases. No significant difference between the mean DNA index and presence of lymph node metastasis was found. These results show the possibility of using flow cytometry for DNA analysis in canine mammary tumors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between DNA ploidy and proliferative cell nuclear antigen index in canine hemangiopericytoma.

The mitotic index is reported to be correlated with recurrence, mean patient survival, and metastasis of canine hemangiopericytoma (CHP). However, to the authors' knowledge, studies investigating the parameters that can predict recurrence or metastasis of CHP with low mitotic index have not been done. To evaluate growth kinetics of CHP with low mitotic index, a retrospective analysis of the proliferative activity by antiproliferative cell nuclear antigen monoclonal antibody and DNA contents by flow cytometry (FCM) was performed with 21 formalin-fixed and paraffin-embedded CHP samples. Of the 21 tumors evaluated by FCM, 6 (26.6%) were aneuploid tumors, and 15 (71.4%) were diploid tumors. There was significant correlation between the PCNA index and ploidy pattern. The diploid group had 39.1 +/- 9.2 PCNA index, whereas the aneuploid group's proliferative cell nuclear antigen (PCNA) index was 63.1 +/- 8.2. The diploid group had mean mitotic index value of 1.140 +/- 0.855, and the aneuploid group had a mean value of 1.067 +/- 0.767. From these results, the CHP samples with low mitotic index were classified into either the aneuploid group with higher PCNA index or the diploid group with lower PCNA index, suggesting that DNA ploidy and proliferative activity may give an indication about malignancy of CHPs with a low mitotic index.     

Clinical and pathologic features of canine ocular melanomas

Ocular melanomas in 40 dogs were reviewed and classified by site of origin and cytologic features. An apparent correlation existed between site, cytologic classification, and prognosis. Limbal melanomas and intraocular melanomas classified as spindle A or spindle B were less aggressive and had a more favorable prognosis than intraocular melanomas classified as a mixed-cell type or epithelioid.     

Prognosis after surgical excision of canine melanomas.

One hundred and thirty-four dogs from which melanomas had been excised were studied until death or for at least 2 years after surgery. Seven of 49 (14%) intraoral and lip tumours and 52 of 85 (61%) skin tumours were histologically benign; in spite of this, three of seven (43%) "benign" oral and four of 52 (8%) "benign" skin lesions led to the eventual death of the host. Thirty eight of 42 (90%) dogs with a histologically malignant melanoma of the lip or oral cavity died because of the tumour but only 15 of 33 (45%) with malignant skin melanomas died. Six of 59 (10%) dogs with a tumour of mitotic index 2 or less died from the tumour 2 years after surgery compared to 19 of 26 (73%) dogs having a tumour with a mitotic index of 3 or more.     

Mast cells and angiogenesis in canine melanomas: malignancy and clinicopathological factors

The biological significance of mast cells and angiogenesis in canine melanomas is unclear. Eighty canine melanomas (56 malignant and 24 benign), investigated to determine the relationship between mast cell count (MCC), microvessel density (MVD) and clinicopathology, revealed significantly higher MCC and MVD counts in malignant melanomas. Evaluation of the prognostic significance of MCC and MVD in malignant melanomas showed a significant correlation between MCC and MVD both within and at the edges of the tumour. Multivariate analysis indicated that MCC and MVD were independent predictors of survival but the former was a significantly better prognostic marker. Greater numbers of mast cells and microvessels were found in malignant melanomas of poor prognosis. The findings demonstrate a prognostic significance of MCC and MVD in canine melanocytic tumours.     

Morphology and behavior of primary ocular melanomas in 91 dogs

Primary ocular melanocytic neoplasms from 91 dogs were divided into two groups by histologic criteria. Seventy-five were benign and composed of spindle-shaped and large polyhedral melanocytes similar to those of human ocular melanocytomas. Fifty-nine of these originated in the uvea where most resulted in uveitis, glaucoma, or hyphema prior to enucleation. None metastasized. Nineteen melanocytomas were limbal tumors. None metastasized, but three of nine incompletely excised tumors were found within the anterior chamber 2 to 3 years after the initial removal. Sixteen uveal melanocytic neoplasms were histologically malignant. Three had confirmed metastases, all within 3 months of enucleation. Cell type or pattern of growth within the globe were not predictive of biologic behavior. Our data suggest that the mitotic index is the best criterion for histologic identification of ocular melanomas with high metastatic potential. We propose that the classification of primary ocular melanomas be simplified to include only two categories: melanocytoma (benign) and melanoma (potentially malignant). Further behavioral data may justify a grading scheme for melanomas based upon mitotic index.     

Cell proliferation and expression of connexins differ in melanotic and amelanotic canine oral melanomas

Melanoma is a malignant neoplasm occurring in several animal species, and is the most frequently found tumor in the oral cavity in dogs. Melanomas are classified into two types: melanotic and amelanotic. Prior research suggests that human amelanotic melanomas are more aggressive than their melanotic counterparts. This study evaluates the behavior of canine melanotic and amelanotic oral cavity melanomas and quantifies cell proliferation and the expression of connexins. Twenty-five melanomas (16 melanotic and 9 amelanotic) were collected from dogs during clinical procedures at the Veterinary Hospital of the School of Veterinary Medicine and Animal Science of the University of São Paulo, Brazil. After diagnosis, dogs were followed until death or euthanasia. Histopathology confirmed the gross melanotic or amelanotic characteristics and tumors were classified according to the WHO. HMB45 or Melan A immunostainings were performed to confirm the diagnosis of amelanotic melanomas. Cell proliferation was quantified both by counting mitotic figures and PCNA positive nuclei. Expressions of connexins 26 and 43 were evaluated by immunohistochemistry, qRT-PCR and Western blot. Dogs bearing amelanotic melanomas presented a shorter lifespan in comparison to those with melanotic melanomas. Cell proliferation was significantly higher in amelanotic melanomas. Expressions of Connexins 26 and 43 were significantly reduced in amelanotic melanomas. The results presented here suggest that oral cavity melanotic and amelanotic melanomas differ regarding their behavior, cell proliferation and connexin expression in dogs, indicating a higher aggressiveness of amelanotic variants.     

Morphology, histology and functional anatomy of the canine cranial cruciate ligament

Morphologically the canine cranial cruciate ligament can be divided into a cranio-medial and caudo-lateral component which perform reciprocal functions at all angles of flexion of the stifle joint. Histologically the main constituents of these two components are bundles of longitudinally orientated collagen fibre. The results of the study of the effect of partial and total sectioning of the cranial cruciate ligament on the "anterior-draw" movement implied that the relatively minor degree of movement, elicited following sectioning of either of the components of the ligament, would not be detected under clinical conditions. For joint instability to be clinically detectable most of the ligament must have ruptured, or the intact portion must have undergone degenerative or disruptive changes.     

Analysis of KIT expression and KIT exon 11 mutations in canine oral malignant melanomas

KIT, a transmembrane receptor tyrosine kinase, is one of the specific targets for anti-cancer therapy. In humans, its expression and mutations have been identified in malignant melanomas and therapies using molecular-targeted agents have been promising in these tumours. As human malignant melanoma, canine malignant melanoma is a fatal disease with metastases and the poor response has been observed with all standard protocols. In our study, KIT expression and exon 11 mutations in dogs with histologically confirmed malignant oral melanomas were evaluated. Although 20 of 39 cases were positive for KIT protein, there was no significant difference between KIT expression and overall survival. Moreover, polymerase chain reaction amplification and sequencing of KIT exon 11 in 17 samples did not detect any mutations and proved disappointing. For several reasons, however, KIT expression and mutations of various exons including exon 11 should be investigated in more cases.     

Immunohistochemical characterization and evaluation of prognostic factors in canine oral melanomas with osteocartilaginous differentiation

Melanomas are the most common malignant oral neoplasm in dogs. Osteocartilaginous differentiation in oral melanomas is a rare feature described both in veterinary and human medicine. Here, 10 cases of this type of neoplasm were used to study their immunohistochemical, biological, and clinical characteristics. Reactivity for S100 and melan A antigen was evaluated, and 4 prognosis factors (mitotic index, invasiveness of epithelium, nuclear atypia, and proliferation index) were analyzed and correlated with the clinical course of the neoplasms after diagnosis. Immunohistochemical analysis of the studied neoplasms, including the osteocartilaginous areas, showed positive immunoreaction for S100 and melan A, except in one dog, which was negative for melan A. Analysis of the results showed that oral melamonas with osteocartilaginous differentiation have a clinical course similar to that of other melanomas in the oral cavity. Analysis of the mitotic index and the expression of proliferation marker Ki-67 could be useful tools for predicting the biological behavior of these neoplasms.     

Biologic behavior of canine uroliths

Canine uroliths may form rapidly or slowly, progressively increase or decrease in size, or become inactive. Associated clinical signs are usually dependent on their locations but may also be influenced by underlying causes. Recurrence of uroliths is unpredictable, being influenced by several variables. Recurrent uroliths are usually, but not invariably, similar in mineral composition to those present during the initial episode.     

Seroconversion of puppies to canine parvovirus and canine distemper virus: a comparison of two combination vaccines

Sixty puppies were randomly assigned to receive one of two commercially available combination vaccines, and responses to the canine parvovirus and canine distemper virus components of the vaccines were determined by measuring serum antibody titers. The percentage of puppies that seroconverted to canine parvovirus was significantly higher and the mean time for seroconversion was significantly shorter for puppies that received one of the vaccines than for puppies that received the other vaccine. Percentages of puppies that seroconverted to canine distemper virus were not significantly different.     

MicroRNAs as tumour suppressors in canine and human melanoma cells and as a prognostic factor in canine melanomas

Malignant melanoma (MM) is one of the most aggressive cancers in dogs and in humans. However, the molecular mechanisms of its development and progression remain unclear. Presently, we examined the expression profile of microRNAs (miRs) in canine oral MM tissues and paired normal oral mucosa tissues by using the microRNA‐microarray assay and quantitative RT‐PCR. Importantly, a decreased expression of miR‐203 was significantly associated with a shorter survival time. Also, miR‐203 and ‐205 were markedly down‐regulated in canine and human MM cell lines tested. Furthermore, the ectopic expression of miR‐205 had a significant inhibitory effect on the cell growth of canine and human melanoma cells tested by targeting erbb3. Our data suggest that miR‐203 is a new prognostic factor in canine oral MMs and that miR‐205 functions as a tumour suppressor by targeting erbb3 in both canine and human MM cells.     

犬的几种社会行为及特征

犬是我们最亲近的朋友,然而我们常常误解它的行为。例如我们通常以为犬是因为内疚或不安才去破坏东西,然而其真正原因却是它受到了惊吓。当犬处在焦虑、     

犬异常攻击行为的分析与预防

一、犬异常攻击行为产生的原因 (一)遗传因素 犬攻击行为的产生是由其防御本能所引发的.防御本能是犬与生俱来的天性,只不过每一头犬,尤其是不同品种的犬存在强弱程度的不同.一般而言,大部分小型宠物犬,其防御本能较弱,有些警用工作犬的防御本能天生就较强,如罗威纳犬、杜伯文犬等.犬具有防御本能,但并不是都会对人产生异常攻击行为.那些容易攻击人的犬的后代攻击性较强.     

Clonality and clinical behavior of canine indolent lymphoma

Introduction: Canine lymphomas with a follicular architecture are biologically indolent and include follicular (FL), mantle cell (MCL), marginal zone (MZL) and T‐zone (TZL) types. They are under recognized and have long survival regardless of treatment. Methods: 70 cases of canine lymphoma with fading follicular hyperplasia (FFH) received¹ over 1996 to present were reviewed. Diagnoses were determined by consensus of three pathologists^1,2 on phenotyped tissues. Clonality for B and T‐cell receptor genes was determined on DNA from paraffin blocks via PCR on 53 of 70 cases². Results: Of the total of 70 cases 52 were of B‐cell type (MZL 44, FL 4, MCL 3, MALT 1) and 9 were of T‐cell type (TZL). 9 cases of benign hyperplasia were included to assist in defining the histologic boundaries of neoplasia. Case presentation included splenic 16, and nodal 54. For 44 cases called MZL on histologic criteria, DNA was available on 33 of which 26 (78.8%) were clonal for B‐cell rearranged gene including 5 also clonal for T‐cell receptor, one clonal for T‐cell only and 6 cases negative for both and considered benign. For TZL, DNA was available on 6 of 9 cases of which 3 were clonal for T‐cell receptor, 1 for B‐cell, 1 for both and 2 polyclonal for both. Conclusions: An Ohio review of 380 cases of canine nodal lymphoma found 29% of cases to be of indolent type as described here and thus a very significant part of the patient population. Seven of 10 cases of MZL treated at Illinois with follow‐up died of other causes than lymphoma at an average of 18.8 months after diagnosis. One of the other three was lost to follow‐up, one was euthanized at owners’ choice at 9.5 m. after diagnosis still in good health and the other was terminated 3 m after diagnosis with concurrent lung carcinoma.     

Investigation of cell-free DNA in canine plasma and its relation to disease

Background: DNA is released from dying cells during apoptosis and necrosis. This cell-free DNA (cfDNA) diffuses into the plasma where it can be measured. In humans, an increase in cfDNA correlates with disease severity and prognosis.

Objective: It was hypothesized that when DNA in canine plasma was measured by emission fluorometry without prior DNA extraction, the concentration of cfDNA would increase with disease severity.

Animals: The diseased population consisted of 97 client-owned dogs. The clinically normal population consisted of nine client-owned dogs presenting for ‘wellness screens’, and 15 colony-owned Harrier Hounds.

Methods: Plasma cfDNA was measured by fluorometry without prior DNA extraction. The effects of ex vivo storage conditions were evaluated in plasma from two clinically normal dogs. In all other dogs, plasma was separated within two hours of collection. The association between the cfDNA concentration in hospitalized dogs and a variety of clinical, clinicopathological and outcome variables was tested.

Results: The concentration of cfDNA was reliably measured when plasma was separated within two hours of blood collection. The diseased dogs had significantly higher cfDNA than clinically normal dogs (P < 0.001), and the more severe the disease, the higher the cfDNA when severity was categorized according to the American Society of Anesthesiologists (ASA) status (P < 0.001). Dogs that did not survive to discharge had significantly higher cfDNA concentrations than survivors (P = 0.02).

Conclusions/Clinical Importance: The concentration of cfDNA in the plasma of diseased dogs is associated with disease severity and prognosis. Measurement of canine cfDNA could be a useful non-specific disease indicator and prognostic tool.