Correlation of cytologic and histopathologic findings with perinodal echogenicity of abdominal lymph nodes in dogs and cats

Abdominal lymphadenopathy in dogs and cats is routinely investigated with ultrasound. As the determination between benign and neoplastic etiologies of lymphadenopathy affects patient management, specific sonographic characteristics associated with both benign and neoplastic lymph nodes have been suggested. However, a significant overlap between these characteristics exists, necessitating a cytologic or histopathologic diagnosis in most instances. The objectives of this retrospective, cross‐sectional study were to evaluate whether echogenicity of perinodal fat could be a discriminator between benign and neoplastic abdominal lymphadenopathy and to assess if additional sonographic features associated with malignancy could be identified in lymph nodes with hyperechoic perinodal fat. Small animal patients (257 dogs and 117 cats) with sonographic evidence of abdominal lymphadenopathy and a cytological or histopathological diagnosis were evaluated for differences in the proportions of sonographic features between benign and neoplastic groups. Greater maximum long axis diameter (in dogs and cats) and a greater number of abnormal lymph nodes (in cats) were associated with malignancy in lymph nodes with hyperechoic perinodal fat. Canine lymph nodes with round cell neoplasia were significantly more likely to have hyperechoic perinodal fat. Lymph nodes affected with other neoplasia or with lymphadenitis were equally likely to have normal or hyperechoic perinodal fat. Reactive lymph nodes were significantly less likely to have hyperechoic perinodal fat in both species. These results suggest that though echogenicity of perinodal fat is a nonspecific finding, abdominal lymph nodes with hyperechoic perinodal fat are less likely to be reactive and sampling of these lymph nodes may be indicated.     

A comparison of the cytologic and histologic features of meningiomas in four dogs

Abstract: The cytologic and histologic features of 2 intracranial and 2 spinal (extramedullary cervical) canine meningiomas were compared. Cerebrospinal fluid analysis in 2 cases revealed mild, mixed cell pleocytosis, primarily composed of small lymphocytes and monocytoid cells, with a moderate increase in total protein concentration. Cytologic features suggestive of meningioma included cells with both epithelial and mes-enchymal characteristics and a tendency towards cell clustering. Tumor location also was useful in making a diagnosis. The 4 meningiomas differed histologically from one another, and included angioblastic, psam-momatous, meningotheliomatous, and microcystic anaplastic types, which conformed to a classification scheme for human meningiomas. The classification scheme could not be applied to cytologic specimens.     

Comparison of B-mode and Doppler ultrasonographic findings with histologic features of benign and malignant superficial lymph nodes in dogs

OBJECTIVE: To compare and correlate B-mode and color Doppler ultrasonographic characteristics with histopathologic findings of benign and malignant superficial lymph nodes in dogs. STUDY POPULATION: 50 superficial lymph nodes that were normal, abnormally large on physical examination, or represented regional lymph nodes draining an area of suspected primary malignancy in 30 dogs. PROCEDURES: Before excision, lymph nodes were evaluated via B-mode and color Doppler ultrasonography to assess size, echogenicity, presence of a hilus, acoustic transmission, and vascular flow. Formalin-fixed, paraffin-embedded tissue sections of excised lymph nodes were stained with H&E and examined for the presence and extent of necrosis, fibrosis, fat, metastases, and tissue heterogeneity. To assess vascularity, the number and distribution of vessels stained by the Verhoeff van Gieson technique were recorded. RESULTS: In superficial lymph nodes, a varied echogenicity corresponded to tissue heterogeneity. The ultrasonographic detection of a hilus was associated with the presence of fibrous tissue, fat, or both in the hilar region. Acoustic enhancement corresponded to presence of areas of intranodal necrosis. There was significant correlation between both the distribution and the number of vessels detected via ultrasonography and that detected by histopathology. The amount of flow estimated via ultrasonography was typically higher than that estimated via histologic examination. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that histopathologic changes in canine lymph nodes have associated ultrasonographic changes and suggest that lymph node ultrasonography has an important role in the evaluation of lymph nodes in dogs in general and in dogs with neoplastic disease in particular.     

Application of rapid CD3 immunophenotype analysis and argyrophilic nucleolar organizer region (AgNOR) frequency to fine needle aspirate specimens from dogs with lymphoma

Determinations of CD3 immunoreactivity (CD3-IR) and argyrophilic nucleolar organizer region (AGNOR) frequency from fine needle aspirate (FNA) samples were compared with those from formalin-fixed, paraffin-embedded surgical biopsy samples in 51 dogs with lymphoma. Both CD3-IR (using a rapid EPOS polymer system) and AGNOR techniques were readily applied to FNA samples. CD3-IR from FNA samples matched those of histologic samples with 100% concordance. A linear relationship (r=0.981; P<0.001) was found between AGNOR frequency obtained from FNA samples and those obtained from surgical biopsy samples. Application of the techniques presented here should allow clinically relevant information to be procured rapidly and inexpensively. As CD3-IR and AGNOR frequency have been shown to be predictive of response to combination chemotherapy in dogs with lymphoma, such information could be used to better educate clients as to the likelihood of achieving meaningful responses, as well as allowing prospective tailoring of individual treatments in future trials prior to initiating therapy.     

Cytochemical staining characteristics of lymph nodes from normal and lymphoma-affected dogs

Frozen sections and imprint smears were used to evaluate the presence and pattern of cytochemical staining reactions in the B- and T-cell regions of lymph nodes from normal dogs and dogs with lymphoma. Staining procedures evaluated included peroxidase (PER), Sudan black B (SBB), naphthol AS-D chloroacetate esterase (CAE), alpha-naphthyl butyrate esterase (NBE), acid phosphatase (ACP), and leukocyte alkaline phosphatase (LAP). In normal lymph nodes, macrophages and some lymphocytes within the interfollicular (T-cell) region and medulla stained positive with ACP and NBE. Smaller numbers of macrophages also occurred sporadically within the germinal follicles. Cells positive for PER, SBB, and CAE were scattered infrequently throughout all regions of the normal lymph node, consistent with granulocytes and mast cells. The LAP stained cells were predominantly and prominently located within the mantle zone of secondary follicles and to a much lesser extent within the germinal centers, compatible with B-cell lymphocytes derived from follicular center cells. Of the 12 dogs with lymphoma, 7 cases (4 immunoblastic, 2 large noncleaved, 1 small noncleaved) stained diffusely positive with LAP, 4 cases (all lymphoblastic) had numerous focally positive lymphocytes using ACP and NBE, and 1 case (immunoblastic) did not stain positive with any of the cytochemical reactions. Cytochemical staining of canine lymph nodes with NBE, ACP, and LAP proved useful in distinguishing between B- or T-cell regions and detecting different cell types of canine lymphoma.     

Comparison of results of clinicians' assessments, cytologic examination of fine-needle lymph node aspirates, and flow cytometry for determination of remission status of lymphoma in dogs

OBJECTIVE: To determine interclinician agreement when assessing remission of lymphoma in dogs and the association among results of clinicians' assessments via lymph node palpation, cytologic examination of fine-needle lymph node aspirates, and flow cytometry as determinants of remission. DESIGN: Prospective study. ANIMALS: 23 dogs with untreated lymphoma. PROCEDURE; Two clinicians independently measured large lymph nodes and cytologic examination and flow cytometry of cells from a mandibular or popliteal lymph node were performed 1 week prior to initiating treatment. Lymph node measurements with clinicians' remission assessments and cytologic examination were repeated at weeks 2, 3, and 5; flow cytometry was repeated at week 5. RESULTS: Significant correlation was identified between clinicians' remission assessments. Significant correlation between lymph node palpation and cytologic examination was identified at week 5, but not at weeks 2 and 3. Lymphoma was diagnosed in 16 of 23 (70%) dogs at initial evaluation by use of flow cytometry, although it was of limited use at subsequent evaluations and results were not diagnostic of lymphoma in any dog at week 5, including 1 dog in which lymphoma was diagnosed cytologically. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that physical examination and measurement of lymph node volume may not be sufficient for accurately determining remission, that flow cytometry alone should not be relied on as a means for diagnosis, and that cytologic examination of fine-needle lymph node aspirates should be considered as the most accurate means of determining remission status at times in which treatment modifications are considered.     

Cestode infection in 2 dogs: cytologic findings in liver and a mesenteric lymph node

Mesocestoides cestode infections in dogs are well known for causing severe peritonitis with larvae or larval fragments (metacestodes, tetrathyridia, or calcareous corpuscles) frequently observed cytologically in peritoneal fluid samples. This case report describes the cytologic and clinical features of 2 dogs infected with cestode larvae, with one case confirmed and the other presumed to be Mesocestoides sp. In these 2 unusual cases, cestode larvae or larval fragments were found in fine‐needle aspirates of the liver and a mesenteric lymph node, but no organisms were found in peritoneal fluid samples. The data presented in this report indicate that clinical pathologists should not rule out Mesocestoides sp cestodiasis based on the absence of larvae in peritoneal fluid samples from dogs.     

Comparisons between elastographic stiffness scores for benign versus malignant lymph nodes in dogs and cats

Lymph node evaluation is an important component of oncology staging in dogs and cats, however diagnosis of malignancy currently requires cytology or histopathology. Elastography is an ultrasound technique that allows estimation of tissue stiffness. In people, it has been shown to increase the sensitivity of sonography in detection of metastatic lymph nodes. The objective of this prospective, cross‐sectional study was to determine if elastographic stiffness differs for malignant versus benign canine and feline lymph nodes that were considered abnormal in gray‐scale ultrasound imaging. Animals scheduled for ultrasound‐guided fine needle aspirates of a lymph node at a single center were prospectively enrolled. Elastography was performed by a board‐certified veterinary radiologist (G.S.) prior to tissue sampling. Softness was scored qualitatively as 1–4 (1 = hard; 4 = soft) on an image depicted by a color scale (blue = hard, red = soft). Quantitative analysis was performed using custom‐made software. Fifty‐one lymph nodes were included in the evaluation, 21 had benign and 30 had neoplastic cytology. Benign lymph nodes were softer (median score of 2.5, range 1–4) than malignant lymph nodes (median = 2, range 1–3), and differences were significant in qualitative and quantitative analyses (P < 0.01). Of the lymph nodes with a score of 2 or below (hard), 19/25 (76%) were malignant, of the lymph nodes with a score over 2 (soft), 24/36 (67%) were benign. Findings indicated that elastographic stiffness scores differed between benign and malignant lymph node groups; however overlapping scores for the two groups limited the use of this technique for routine clinical diagnosis of malignancy.     

Radiographic characterization of enlarged sternal lymph nodes in 71 dogs and 13 cats

In this retrospective study, radiographically enlarged sternal lymph nodes (LNs) were evaluated in 71 dogs and 13 cats for average size, location, and most representative radiographic view. Concurrent clinical diagnoses were also noted and grouped into one of three following categories: neoplastic, inflammatory, or hematologic. There were no statistically significant differences in LN size between lateral views within each species. Enlarged sternal LNs were more cranially positioned in dogs than cats. No statistical difference was noted between right and left laterals, as to on which projection the enlarged sterna lymph nodes was seen best. Neoplastic disease (78.9%) was the most prevalent condition seen in association with LN enlargement in dogs, followed by primary infectious or inflammatory diseases (14.1%) and various hematologic conditions (7.0%). In cats, neoplasia was also most common (69.2%), followed by inflammatory diseases (30.8%). No hematologic conditions were noted in cats. The most common etiologic agent seen concurrently with enlarged sternal LNs in both dogs (33.8%) and cats (38.5%) was malignant lymphoma. The results of this study provide a clinically useful representation of the average size and location of radiographically enlarged sternal LNs for dogs and cats. The diseases represented demonstrate the wide spectrum of potential causes of sternal lymphadenopathy.     

Correlation between histologic diagnosis mean nucleolar organizer region count and prognosis in canine mammary tumors.

In this study, surgically excised mammary tumors from 98 bitches were graded histologically, and the grade was compared with the mean nucleolar organizer region (NOR) count in silver-stained paraffin-embedded sections. Histologically benign tumors, papillary adenocarcinomas, and intraductal carcinomas showed relatively little variation; the mean count for each category was between three and four NOR per nucleus. There was, however, a significant increase in the NOR counts in tubular and solid carcinomas. This increase was most pronounced for tumors that showed evidence of infiltration into the surrounding connective tissues. The mean NOR count for noninfiltrative carcinomas was 5.1, and that for invasive carcinomas was 7.3 (P less than 0.03). The mean NOR count for individual carcinomas ranged from 2.0 to 12.3, and a significant correlation was found between an increased NOR count and tumor-related death during the first post-surgical year. The 39 bitches in which the tumor had an NOR count less than 8.0 had a generally favorable prognosis; only six (15%) died as a result of the original neoplasm. In contrast, 18/21 dogs (85%) with a carcinoma having an NOR count greater than 8.0 died from the tumor during the first post-surgical year. A similar, although less pronounced result was obtained specifically for invasive carcinomas, in which 3/12 (25%) tumors with an NOR count less than 6.0 resulted in the death of the host, compared with 17/20 (85%) that had an NOR count greater than 6. By using this technique, it is possible to identify a subgroup of bitches with invasive mammary carcinomas that have a very poor prognosis following apparently adequate surgical ablation of the primary tumor.     

Sonographic characteristics of presumptively normal main axillary and superficial cervical lymph nodes in dogs

Objective-To evaluate the B-mode and Doppler ultrasonographic appearance of presumptively normal main axillary and large superficial cervical lymph nodes (MALNs and SCLNs, respectively) in adult dogs. Animals-51 healthy adult dogs (data from 1 dog were not analyzed). Procedures-For each dog, weight, distance from the cranial aspect of the first sternebra to the caudal aspect of the left ischiatic tuberosity, and thoracic height and width at the level of the xiphoid process were recorded. Via B-mode and Doppler ultrasonography, echogenic characteristics, size in relation to body size and weight, and vascular supply of the MALNs and the SCLNs were evaluated (1 SCLN in 1 dog was not ultrasonographically visible). Results-Most MALNs were clearly margined, solitary, and ovoid; echopatterns were homogenous or cortical and hypo- to isoechoic, compared with surrounding soft tissues. Size measurements of MALNs correlated with dogs' body length, thoracic width and height, and body weight. Most SCLNs were clearly margined, fusiform, and hypoechoic (compared with surrounding soft tissues) with a cortical or homogenous echopattern. Size measurements of SCLNs correlated with dogs' body length, thoracic width and height, and body weight. In 50 of the 100 MALNs, an intranodal vascular supply was detected; in contrast, an intranodal vascular supply in SCLNs was detected infrequently. Conclusions and Clinical Relevance-Results indicated that, in dogs, anatomically separate lymph nodes have different echogenic and vascular characteristics; body size (skeletal length, height, and width), along with body weight, were correlated with sizes of presumptively normal MALNs and SCLNs.     

Association of argyrophilic nucleolar organizing regions, Ki-67, and proliferating cell nuclear antigen scores with histologic grade and survival in dogs with soft tissue sarcomas: 60 cases (1996-2002)

OBJECTIVE: To determine whether argyrophilic nucleolar organizing regions (AgNORs), Ki-67, and proliferating cell nuclear antigen (PCNA) scores were associated with histologic grade and survival in dogs with soft tissue sarcomas (STSs). DESIGN: Retrospective study. ANIMALS: 60 dogs with STSs. PROCEDURE: Medical records were examined and histologic specimens were reviewed. Tissue specimens obtained from archival materials were used to prepare sections for histologic staining for AgNOR and immunohistochemical staining for Ki-67 and PCNA labeling. Follow-up monitoring was obtained by reevaluation or telephone conversations with referring veterinarians or owners. RESULTS: 27 (45%) STSs were grade 1, 23 (38%) were grade 2, and 10 (17%) were grade 3. The mean and median AgNOR, Ki-67, and PCNA scores were determined, and significant positive associations among AgNOR and Ki-67 scores with histologic grade and mitotic score were detected. Fifty-four dogs had adequate follow-up examinations and were included in survival analysis and evaluation of prognostic factors. Overall median survival time was > 1,306 days. Twelve of 54 (22%) dogs died of tumor-related causes. Metastatic disease developed in 8 of 54 (15%) dogs. Results of univariate analysis indicated that increased mitotic score, increased AgNOR score, increased Ki-67 score, incomplete surgical margins, noncurative intent surgery, Ki-67 score greater than the median Ki-67 score, and AgNOR score greater than the median AgNOR score were prognostic factors for decreased survival time. Results of multivariate analysis indicated that increased AgNOR score was the only prognostic factor for decreased survival time. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that AgNORs and possibly Ki-67 should be routinely evaluated with histologic grading for STSs in dogs.     

Lymphotropic nanoparticle magnetic resonance imaging for diagnosing metastatic lymph nodes in dogs with malignant head and neck tumours

Lymphotropic nanoparticle magnetic resonance imaging (LNMRI) utilises ultrasmall paramagnetic iron nanoparticles (USPIOs) for imaging of metastatic lymph nodes in patients afflicted with cancer. LNMRI has been shown to be a highly effective and accurate way to diagnose metastasis in humans but has not been commonly reported on in veterinary medicine. USPIOs are phagocytised by macrophages and then localised to lymph nodes where they create a susceptibility artefact on gradient echo MRI sequences. In this study dogs (n = 24) with naturally occurring head and neck tumours were imaged with LNMRI then had mandibular and retropharyngeal lymph nodes extirpated for histological analysis. Subjective and objective analysis of the LNMRI images was performed and imaging results compared to histology as the gold standard. A total of 149 lymph nodes were included in this study. The overall sensitivity, specificity and accuracy was 64%, 94.4% and 89.3% respectively. However, if dogs with mast cell tumours were excluded from analysis the sensitivity, specificity and accuracy rose to 85.7%, 95.7% and 94.6%. LNMRI is potentially an accurate way to determine the presence of lymph node metastasis in dogs with some types of head and neck tumours. However, LNMRI has only moderate accuracy in dogs with oral or mucocutaneous mast cell tumours in this region.     

Scanning electron microscopy of vascular corrosion casts and histologic examination of pulmonary microvasculature in dogs with dirofilariosis

OBJECTIVE: To characterize structural changes in pulmonary vessels of dogs with dirofilariosis. ANIMALS: 8 dogs with dirofilariosis and 2 unaffected control dogs. PROCEDURE: Pulmonary artery pressure was measured in affected dogs, and dogs then were euthanatized. Scanning electron microscopy was used to examine vascular corrosion casts of pulmonary vasculature. Tissue sections of pulmonary vasculature were evaluated by use of histologic examination. RESULTS: Pulmonary artery pressure was higher in dogs with severely affected pulmonary vessels. In tissue sections, dilatation, as well as lesions in the tunica intima and proliferative lesions resulting in constriction or obstruction, were frequently observed in branches of the pulmonary artery. Numerous dilated bronchial arteries were observed around affected pulmonary arteries. Hyperplastic venous sphincters were observed in small pulmonary veins and venules. In corrosion casts, affected pulmonary lobar arteries had dilatation, pruning, abnormal tapering, constriction, and obstruction. In small arteries and arterioles, surface structures representing aneurisms and edema were seen. Bronchial arteries were well developed and extremely dilated, and they formed numerous anastomoses with pulmonary arteries at all levels, from the pulmonary trunk to peripheral vessels. Capillaries in the lungs were dilated with little structural change. Small pulmonary veins and venules had irregular annular constrictions that were caused by hyperplastic smooth muscle cells of venous sphincters. CONCLUSIONS AND CLINICAL RELEVANCE: Scanning electron microscopy of microvascular casts delineated links between the bronchial and pulmonary circulations in dogs with dirofilariosis. Results of scanning electron microscopy provided a structural explanation for the development of pulmonary circulatory disturbances and pulmonary hypertension in dogs affected by dirofilariosis.     

Sensitivity and specificity of methods of assessing the regional lymph nodes for evidence of metastasis in dogs and cats with solid tumors

OBJECTIVE: To determine sensitivity and specificity of physical examination, fine-needle aspiration, and needle core biopsy of the regional lymph nodes for evidence of metastasis in dogs and cats with solid tumors. DESIGN: Case series. ANIMALS: 37 dogs and 7 cats. PROCEDURE: Regional lymph nodes were evaluated by means of physical examination (palpation), fine-needle aspiration, and needle core biopsy. Results were compared with results of histologic examination of the entire lymph node, the current standard. RESULTS: Tumors included 18 sarcomas, 16 carcinomas, 7 mast cell tumors, and 3 other tumors. Carcinomas were more likely to have metastasized to the regional lymph node (7/16 animals) than were sarcomas (2/18). Sensitivity and specificity of physical examination were 60 and 72%, respectively. Sensitivity and specificity of cytologic examination of fine-needle aspirates were 100 and 96%, respectively. Sensitivity and specificity of histologic examination of needle core biopsy specimens were 64 and 96%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that fine-needle aspiration may be a sensitive and specific method of evaluating the regional lymph nodes in dogs and cats with solid tumors, because results correlated well with results of histologic examination of the entire lymph node. Physical examination alone was not a reliable method and should not be used to decide whether to aspirate or biopsy the regional lymph nodes.