Association between cigarette smoke exposure and urinary bladder cancer in Scottish terriers in a cohort study.

Canine urothelial carcinoma (UC) initially responds favorably to treatment, but is ultimately lethal in most cases. Research to identify modifiable risk factors to prevent the cancer is essential. The high breed-associated risk for UC, e.g. 20-fold higher in Scottish terriers, can facilitate this research. The objective was to identify environmental and host factors associated with UC in a cohort of Scottish terriers. Information was obtained through dog owner questionnaires for 120 Scottish terriers ≥ 6 years old participating in a bladder cancer screening study, with comparisons made between dogs that did or did not develop UC during the 3 years of screening. Univariable models were constructed, and variables with P < 0.20 were included when building the multivariable model, and then removed using a backward stepwise procedure. P < 0.05 was considered statistically significant. Urine cotinine concentrations were measured by liquid chromatography-mass spectrometry to further investigate potential cigarette smoke exposure. Biopsy-confirmed UC which was found in 32 of 120 dogs, was significantly associated with the dogs living in a household with cigarette smokers (odds ratio [OR], 6.34; 95 % confidence intervals [CI], 1.16-34.69; P = 0.033), living within a mile of a marsh or wetland (OR, 21.23; 95 % CI, 3.64-123.69; P = 0.001), and history of previous bladder infections (OR, 3.87; 95 % CI, 1.0-14.98; P = 0.050). UC was diagnosed in 18 of 51 dogs (35.3 %) with quantifiable cotinine concentrations, and six of 40 dogs (15.0 %) without quantifiable cotinine concentrations in their urine (P = 0.0165). In conclusion, the main modifiable risk factor for UC in this cohort of dogs was exposure to second-hand tobacco smoke.

Retrospective analysis of factors affecting clinical outcome following CHOP-based chemotherapy in dogs with primary nodal diffuse large B-cell lymphoma.

Numerous factors are known to affect the prognosis of dogs with chemotherapy-treated lymphomas. However, prognostic factors for dogs with specific subtypes of lymphoma are less clearly defined. The objective of this study was to identify prognostic factors for dogs receiving CHOP-based chemotherapy for primary nodal diffuse large B-cell lymphoma (DLBCL). Medical records of dogs treated for DLBCL at the Purdue Veterinary Teaching Hospital (PUVTH) from 2006 to 2016 were reviewed. Factors potentially related to prognosis were analysed using multivariable statistical methods. Ninety-eight dogs were included in the study. Best overall response to chemotherapy was complete remission in 80 dogs (81.6%) and partial remission in 18 dogs (18.4%). Median progression-free survival (PFS) for the entire population was 252 days (range 19-1068). Factors significantly associated with achieving partial (rather than complete) remission following CHOP included presence of thrombocytopenia at diagnosis (OR 6.88; 95% CI 1.98-23.93; P = .002), baseline serum globulin concentration (OR 2.63; 95% CI 1.03-6.75; P = .044), and age at diagnosis (OR 1.36; 95% CI 1.08-1.71; P = .009). Factors significantly associated with PFS in the lowest quartile (≤93 days) included presence of thrombocytopenia at diagnosis (OR 8.72; 95% CI 1.54-49.33; P = .014), age at diagnosis (OR 1.47; 95% CI 1.12-1.94; P = .005), and baseline neutrophil count (OR 1.18; 95% CI 1.02-1.37; P = .025). Presence of thrombocytopenia, greater age, higher neutrophil count, and higher serum globulin concentration all may be associated with a particularly poor outcome in dogs receiving CHOP-based chemotherapy for DLBCL.

Immunohistochemical Profile of 20 Feline Renal Cell Carcinomas.

Renal cell carcinoma (RCC) is uncommon in cats, but makes up the majority of epithelial neoplasms in the kidney. The immunohistochemical profile of 20 feline RCCs (13 tubular carcinomas, four tubulopapillary carcinomas, one papillary carcinoma and two anaplastic carcinomas) was evaluated. Primary antibodies used were specific for Pax8, KIT, CD10, cytokeratins and vimentin. A polymer-based immunoperoxidase procedure was used. Nineteen tumours (95%) expressed Pax8; 12 (60%), KIT; 15 (75%), CD10; 20 (100%), cytokeratins; and 19 (95%), vimentin. Nuclear Pax8 immunoreactivity was readily apparent, but variation in labelling intensity was present within a given section. KIT reactivity was diffuse, cytoplasmic and relatively homogeneous. CD10 immunoreactivity was predominantly membranous along the apical border of tubular epithelial cells and was less commonly cytoplasmic. CD10 immunoreactivity was less intense in areas with papillary differentiation and absent in solid areas. Cytoplasmic cytokeratin expression was strong in 18 tumours and weak in two; the papillary portion of one tumour had distinct submembranous expression. Vimentin immunoreactivity, which ranged from diffuse to focal, was difficult to evaluate due to strong stromal immunoreactivity and its patchy expression in phenotypically similar neoplastic cells. Fewer non-renal tumours were positive for Pax8 than for CD10. Considering overall sensitivity and specificity, Pax8 appears to be a valuable marker for distinguishing feline tumours arising in the kidney from other neoplasms.

Immunohistochemical Detection of Pax8 and Napsin A in Canine Thyroid Tumours: Comparison with Thyroglobulin, Calcitonin and Thyroid Transcription Factor 1.

Expression of thyroid transcription factor (TTF)-1 corroborates a thyroid origin of neoplasms. Thyroglobulin and calcitonin immunohistochemistry (IHC) can distinguish between a follicular and C-cell origin of thyroid tumours, respectively. Pax8 (expressed by normal canine thyroid follicular cells) and napsin A (expressed mainly by C-cells) labelling was compared with labelling for TTF-1, thyroglobulin and calcitonin in 114 canine proliferative thyroid lesions. All 81 follicular tumours expressed thyroglobulin and were negative for calcitonin; 79/81 (98%) of these tumours expressed TTF-1 and Pax8 and 60/81 (74%) expressed napsin A. All 25 C-cell lesions expressed calcitonin and were negative for expression of thyroglobulin; 22 (88%) were positive for TTF-1, 13 (57%) for Pax8 and 24/24 for napsin A. Six mixed follicular-medullary carcinomas expressed all five markers. Both carcinosarcomas expressed TTF-1 and napsin A, and one each of these tumours expressed thyroglobulin, calcitonin or Pax8. Pax8 expression was also detected in epididymal cells, endometrial cells and vas deferens epithelium, in Sertoli-like ovarian cells, and in some cases of ovarian adenoma, pancreatic carcinoma, renal cell carcinoma and Sertoli cell tumour. Napsin A was also detected in adrenocortical cells, ovarian granulosa cells, epididymal and endometrial cells, as well as in some renal cell carcinomas, pulmonary adenocarcinomas and Sertoli cell tumours. In summary, Pax8 was as sensitive as TTF-1 and slightly less sensitive than thyroglobulin for identification of follicular tumours, but had low sensitivity for C-cell tumours. Napsin A was as sensitive as calcitonin for C-cell neoplasms, but was less sensitive than thyroglobulin for follicular neoplasms. Thus, these markers are sensitive and, except for renal cell carcinoma (for Pax8, napsin A) and pulmonary adenocarcinoma (for napsin A), are specific thyroid tumour markers.

Squamous Papillomas of the Conjunctiva in Dogs: A Condition Not Associated With Papillomavirus Infection.

Papillomas of the conjunctival surface in people can be of viral or nonviral origin and are found in high association with human papillomavirus. Canine conjunctival papillomas are seldom described, and published accounts have mostly been associated with canine oral papillomavirus infection. Here, we describe conjunctival squamous papillomas that do not express papillomavirus proteins and compare them with papillomavirus-associated conjunctival papillomas. Conjunctival squamous papillomas presented a distinct histopathologic profile and lacked the cytopathic effects seen in viral papillomas. They appeared as exophytic, papilliferous, pedunculated lesions with delicate fronds and angular terminal margins. Squamous papillomas presented with a delicate fibrovascular core and were associated both clinically and grossly with a feeder vessel. Pigmentation was variable within the epithelium and stroma of these lesions, and inflammatory infiltrates were characteristically minimal. Conjunctival squamous papillomas resembled squamous papillomas of the skin; however, they lacked significant hyperkeratosis. Compared with conjunctival viral papillomas, these masses occurred in older dogs and were smaller and solitary. Furthermore, polymerase chain reaction and immunohistochemistry failed to demonstrate papillomavirus genetic material and antigens in conjunctival squamous papillomas. Both viral and nonviral conjunctival papillomas were considered benign.

When tissue antigens and antibodies get along: revisiting the technical aspects of immunohistochemistry--the red, brown, and blue technique.

Once focused mainly on the characterization of neoplasms, immunohistochemistry (IHC) today is used in the investigation of a broad range of disease processes with applications in diagnosis, prognostication, therapeutic decisions to tailor treatment to an individual patient, and investigations into the pathogenesis of disease. This review addresses the technical aspects of immunohistochemistry (and, to a lesser extent, immunocytochemistry) with attention to the antigen-antibody reaction, optimal fixation techniques, tissue processing considerations, antigen retrieval methods, detection systems, selection and use of an autostainer, standardization and validation of IHC tests, preparation of proper tissue and reagent controls, tissue microarrays and other high-throughput systems, quality assurance/quality control measures, interpretation of the IHC reaction, and reporting of results. It is now more important than ever, with these sophisticated applications, to standardize the entire IHC process from tissue collection through interpretation and reporting to minimize variability among laboratories and to facilitate quantification and interlaboratory comparison of IHC results.

Immunohistochemical evaluation of the effects of paraffin section storage on biomarker stability.

Environmental stresses can alter immunoreactivity of biomarkers in stored tissue sections. The effect of temperature and lighting on 49 cellular or microbial antigens was evaluated in 4 serial paraffin sections, cut 12 months, 10 months, 8 months, 5 months, 3 months, 1 month, 3 days, and 1 day before immunohistochemistry. Slides were stored at room temperature (RT) in the dark, at 4°C in the dark, at RT under fluorescent light, or at RT with windowpane exposure to sunlight. Immunohistochemistry was performed simultaneously in an automated immunostainer. Immunoreactivity was compared with that in the corresponding 1-day-old section and scored as 4 (<10% reduction), 3 (10%-25% reduction), 2 (26%-60% reduction), 1(>60% reduction), or 0 (no reactivity). Any loss of immunoreactivity was proportional to the tissue section age and was least in sections stored in the dark. Immunoreactivity was only completely lost in light-exposed sections and as early as 1 month for CD45. Other markers with complete loss of immunoreactivity were bovine viral diarrhea virus, CD18 (only with fluorescent light), CD31, CD68, canine parvovirus, chromogranins, and thyroid transcription factor-1. Markers with complete loss after light exposure also had reduced immunoreactivity when stored in the dark, as early as day 3. Eight markers (Bartonella spp, CD11d, high molecular weight cytokeratins, feline coronavirus, GATA-4, insulin, p63, progesterone receptor) had minimal decrease in immunoreactivity, regardless of treatment. In conclusion, light-induced antigen decay (tissue section aging) is antigen dependent and could explain unexpectedly weak or negative immunohistochemical reactions in stored paraffin sections.

Immunohistochemical expression of melanocytic antigen PNL2, Melan A, S100, and PGP 9.5 in equine melanocytic neoplasms.

The immunoreactivity of PNL2, Melan A, and protein gene product (PGP) 9.5 was compared with that of S100 protein in 50 formalin-fixed, paraffin-embedded equine melanocytic neoplasms. PNL2, PGP 9.5, and S100 protein were detected in all 50 neoplasms; none expressed Melan A. PNL2 was not expressed in 62 nonmelanocytic tumors (equine sarcoids, schwannomas, carcinomas, sarcomas, endocrine tumors, sex-cord stromal tumors, germ cell tumors, and leukocytic tumors) or in normal tissues other than epidermis. In summary, antibody PNL2 is a sensitive marker of equine melanocytic neoplasms and is more specific than S100 protein or PGP 9.5. In contrast, the monoclonal antibody to Melan A did not react with any of the equine melanomas.

Randomized trial of cisplatin versus firocoxib versus cisplatin/firocoxib in dogs with transitional cell carcinoma of the urinary bladder.

BACKGROUND: Cisplatin combined with a nonselective cyclooxygenase (cox) inhibitor has potent antitumor activity against transitional cell carcinoma (TCC) in dogs, but this treatment is limited by renal toxicosis. Cox-2 is expressed in TCC, but only in limited sites within the kidney. A cox-2 inhibitor could enhance the antitumor activity of cisplatin with potentially fewer adverse effects on the kidney. HYPOTHESIS: Cisplatin/cox-2 inhibitor treatment will have greater antitumor activity but no more renal toxicosis than cisplatin alone in dogs with TCC. ANIMALS: Forty-four dogs with naturally occurring urinary bladder TCC. METHODS: Dogs were randomized to receive cisplatin (60 mg/m(2) IV q21d), firocoxib (5 mg/kg PO q24h), or the combination. Tumor measurements were determined before and at 6-week intervals during treatment. Renal function was monitored by serum creatinine concentration, iohexol clearance, and urine specific gravity. Toxicoses were graded according to Veterinary Co-Operative Oncology Group (VCOG) criteria. RESULTS: The remission rate with cisplatin/firocoxib (57%) was significantly (P = .021) higher than that with cisplatin alone (13%). Renal and gastrointestinal toxicoses were common in dogs receiving cisplatin, but there were no significant differences between dogs receiving cisplatin or cisplatin/firocoxib. Firocoxib alone induced partial remission or stable disease in 20 and 33% of dogs, respectively. CONCLUSIONS: Firocoxib significantly enhanced the antitumor activity of cisplatin resulting in partial remission in more than half of the cases. The toxicoses inherent to cisplatin, however, were noted in dogs receiving this combination. Firocoxib had antitumor effects as a single agent and can be considered a palliative treatment for dogs with TCC.

Gonadoblastoma in the ovaries of a lesser galago (Galago senegalensis braccatus).

An enlarged right ovary was removed from a 14-year-old lesser galago (Galago senegalensis braccatus). Cytological preparations consisted of a heterogeneous population of neoplastic cells admixed with extracellular hyaline structures and cell-free nuclei. Microscopically, the ovary was replaced with gonadoblastoma and was composed of nests of germinal cells, including large oocyte-like cells, and sex cord-stromal cells arranged in palisading patterns around the germinal cells, the periphery of the nests and around extracellular hyaline material. The animal died 2 years after initial diagnosis. Necropsy examination revealed gonadoblastoma in the left ovary. The germinal cells of the tumour in the right and left ovaries were immunoreactive for calretinin, OCT3/4, PGP 9.5, Ki67 and/or faintly for cytokeratins. Sex cord-stromal cells were immunoreactive for calretinin, OCT3/4, GATA-4, E-cadherin and vimentin. Luteinized sex cord-stromal cells were immunoreactive for inhibin-alpha. The extracellular hyaline material was immunoreactive for laminin. This is the first case of gonadoblastoma in a non-human primate.

Immunohistochemical characterization of nonhuman primate ovarian sex cord-stromal tumors.

This study evaluates the immunoreactivity of 12 sex cord-stromal tumors of nonhuman primates (11 granulosa cell tumors and 1 luteoma). The markers selected are used in the characterization of gonadal tumors in dogs and other species, including cytokeratins AE1/AE3, GATA-4, inhibin-α, neuron-specific enolase, protein gene product 9.5, and vimentin. A normal nonhuman primate ovary was used as a control and to optimize immunolabeling. Staining was graded as follows: 0 (nonstaining), 1+ (< 10% positive cells), 2+ (10%-50% positive cells), and 3+ (> 50% positive cells). Calretinin, GATA-4, neuron-specific enolase, and vimentin were the most consistently expressed markers (12 of 12). Cytokeratins AE1/AE3 were also consistently expressed (11 of 12). Inhibin-α and protein gene product 9.5 were expressed in 8 and 10 sex cord-stromal tumors, respectively. Results indicate that immunoreactivity of nonhuman primate sex cord-stromal tumors is similar to that observed in other species and that calretinin, GATA-4, and neuron-specific enolase are the most consistently expressed markers in nonhuman primate sex cord-stromal tumors.

Comparison of CD34, CD31, and factor VIII-related antigen immunohistochemical expression in feline vascular neoplasms and CD34 expression in feline nonvascular neoplasms.

The diagnosis of vascular neoplasms is often facilitated by the use of immunohistochemical markers such as factor VIII-related antigen, CD31, and CD34. However, the relative sensitivity and specificity of these markers have not been compared in cat vascular neoplasms. In this study, these 3 immunohistochemical markers were evaluated in 61 endothelial neoplasms (50 hemangiosarcomas and 11 hemangiomas) in 59 cats. All neoplasms were labeled by all 3 markers. CD34 had the highest average immunolabeling intensity in neoplastic endothelial cells. CD31 had the lowest average background labeling, followed by CD34 and factor VIII-related antigen, respectively. CD34 expression was also examined in 130 nonvascular neoplasms of cats; 14 of 62 epithelial neoplasms, 39 of 43 mesenchymal neoplasms, 8 of 23 leukocytic neoplasms, and 2 of 2 melanomas were positive. Given the broad expression of CD34 in mesenchymal neoplasms, this marker has limited diagnostic relevance for vascular neoplasms of cats.

Immunohistochemical expression of E-cadherin does not distinguish canine cutaneous histiocytoma from other canine round cell tumors.

Immunohistochemistry for E-cadherin (ECAD) has been used to distinguish canine cutaneous histiocytoma from other leukocytic neoplasms ("round cell tumors"). To determine the specificity of this test, 5 types of canine cutaneous round cell tumors were evaluated for immunohistochemical expression of ECAD. Tumors of all 5 types had variable cytoplasmic, plasma membrane, and/or paranuclear ECAD expression: All 13 cutaneous histiocytomas were ECAD+; all but 1 of 14 mast cell tumors expressed ECAD; 10 of 12 epitheliotropic lymphomas reacted with E-cadherin antibody; of 72 plasmacytomas, 54 were ECAD+; and 5 of 5 histiocytic sarcomas were positive. Conclusions based on these results include the following: First, immunoreactivity for ECAD is not limited to leukocytes of cutaneous histiocytoma; second, antibody to ECAD also labels neoplastic cells in most mast cell tumors, plasmacytomas, cutaneous histiocytic sarcomas, and epitheliotropic lymphomas; third, although most histiocytomas have membranous ECAD expression, the immunoreactivity varies among round cell tumors and is frequently concurrent in different cellular compartments; fourth, the distinctively paranuclear ECAD expression pattern in epitheliotropic lymphomas might distinguish them from other round cell tumors; and, fifth, ECAD should be used with other markers (eg, MUM1 for plasmacytomas, KIT for mast cell tumors, CD3 and CD79a for lymphomas) to distinguish among canine round cell tumors.

Proposal of a 2-tier histologic grading system for canine cutaneous mast cell tumors to more accurately predict biological behavior.

Currently, prognostic and therapeutic determinations for canine cutaneous mast cell tumors (MCTs) are primarily based on histologic grade. However, the use of different grading systems by veterinary pathologists and institutional modifications make the prognostic value of histologic grading highly questionable. To evaluate the consistency of microscopic grading among veterinary pathologists and the prognostic significance of the Patnaik grading system, 95 cutaneous MCTs from 95 dogs were graded in a blinded study by 28 veterinary pathologists from 16 institutions. Concordance among veterinary pathologists was 75% for the diagnosis of grade 3 MCTs and less than 64% for the diagnosis of grade 1 and 2 MCTs. To improve concordance among pathologists and to provide better prognostic significance, a 2-tier histologic grading system was devised. The diagnosis of high-grade MCTs is based on the presence of any one of the following criteria: at least 7 mitotic figures in 10 high-power fields (hpf); at least 3 multinucleated (3 or more nuclei) cells in 10 hpf; at least 3 bizarre nuclei in 10 hpf; karyomegaly (ie, nuclear diameters of at least 10% of neoplastic cells vary by at least two-fold). Fields with the highest mitotic activity or with the highest degree of anisokaryosis were selected to assess the different parameters. According to the novel grading system, high-grade MCTs were significantly associated with shorter time to metastasis or new tumor development, and with shorter survival time. The median survival time was less than 4 months for high-grade MCTs but more than 2 years for low-grade MCTs.

Recommended guidelines for submission, trimming, margin evaluation, and reporting of tumor biopsy specimens in veterinary surgical pathology.

Neoplastic diseases are typically diagnosed by biopsy and histopathological evaluation. The pathology report is key in determining prognosis, therapeutic decisions, and overall case management and therefore requires diagnostic accuracy, completeness, and clarity. Successful management relies on collaboration between clinical veterinarians, oncologists, and pathologists. To date there has been no standardized approach or guideline for the submission, trimming, margin evaluation, or reporting of neoplastic biopsy specimens in veterinary medicine. To address this issue, a committee consisting of veterinary pathologists and oncologists was established under the auspices of the American College of Veterinary Pathologists Oncology Committee. These consensus guidelines were subsequently reviewed and endorsed by a large international group of veterinary pathologists. These recommended guidelines are not mandated but rather exist to help clinicians and veterinary pathologists optimally handle neoplastic biopsy samples. Many of these guidelines represent the collective experience of the committee members and consensus group when assessing neoplastic lesions from veterinary patients but have not met the rigors of definitive scientific study and investigation. These questions of technique, analysis, and evaluation should be put through formal scrutiny in rigorous clinical studies in the near future so that more definitive guidelines can be derived.

Immunohistochemical identification of canine melanocytic neoplasms with antibodies to melanocytic antigen PNL2 and tyrosinase: comparison with Melan A.

The immunoreactivity of PNL2 and antityrosinase in formalin-fixed, paraffin-embedded canine melanocytic neoplasms (n = 101) was compared with that of Melan A. Of the 113 samples overall, 106 were positive for PNL2, 101 for Melan A, and 90 for tyrosinase. Six melanomas that were positive for PNL2 were negative for Melan A; 1 melanoma that was negative for PNL2 was positive for Melan A. Eighty tumors were positive for all 3 markers; 111 reacted with at least 1 the 3 antibodies. Decalcification with formic acid for up to 1 week did not affect immunoreactivity of any of the markers; however, decalcification with HCl for 1 day or 1 week notably decreased or completely abrogated immunoreactivity for Melan A and PNL2. There was only minor loss of immunoreactivity for tyrosinase in tissues decalcified with HCl for 1 week. Prolonged fixation (up to 2 months) did not affect PNL2 or tyrosinase immunoreactivity; however, Melan A immunoreactivity was reduced after 1 month of fixation. PNL2 was not expressed in 120 nonmelanocytic tumors (carcinomas, sarcomas, steroid-producing tumors, and leukocytic tumors). In summary, antibody PNL2 is slightly more sensitive than Melan A and more sensitive than tyrosinase in the identification of canine melanocytic neoplasms. Furthermore, PNL2 does not appear to cross-react with nonmelanocytic neoplasms. PNL2 is resistant to prolonged fixation but sensitive to strong decalcification. Results indicate that PNL2 is an excellent marker in the identification of canine melanomas and that the sensitivity is close to 100% when used in conjunction with Melan A and tyrosinase.

Classification of canine malignant lymphomas according to the World Health Organization criteria.

A study was carried out to test the accuracy and consistency of veterinary pathologists, not specialists in hematopathology, in applying the World Health Organization (WHO) system of classification of canine lymphomas. This study represents an initiative of the ACVP Oncology Committee, and the classification has been endorsed by the World Small Animal Veterinary Association (WASVA). Tissue biopsies from cases of canine lymphoma were received from veterinary oncologists, and a study by pathologists given only signalment was carried out on 300 cases. Twenty pathologists reviewed these 300 cases with each required to choose a diagnosis from a list of 43 B and T cell lymphomas. Three of the 20 were hematopathologists who determined the consensus diagnosis for each case. The 17 who formed the test group were experienced but not specialists in hematopathology, and most were diplomates of the American or European Colleges of Veterinary Pathology. The overall accuracy of the 17 pathologists on the 300 cases was 83%. When the analysis was limited to the 6 most common diagnoses, containing 80% of all cases, accuracy rose to 87%. In a test of reproducibility enabled by reintroducing 5% of cases entered under a different identity, the overall agreement between the first and second diagnosis ranged from 40 to 87%. The statistical review included 43,000 data points for each of the 20 pathologists.

Immunohistochemical detection of CD34, E-cadherin, claudin-1, glucose transporter 1, laminin, and protein gene product 9.5 in 28 canine and 8 feline meningiomas.

The variation in histologic pattern of meningiomas can make their diagnosis challenging. The immunohistochemical profile of 28 canine and 8 feline meningiomas was examined. Tumor types included anaplastic (6 dogs), angiomatoid (1 cat), fibroblastic (3 dogs, 1 cat), meningothelial (1 dog), microcystic (2 dogs), myxoid (3 dogs), psammomatous (4 cats), and transitional (13 dogs, 2 cats). The authors compared the expression of novel markers (CD34, E-cadherin, claudin-1, glucose transporter 1 [GLUT-1], laminin, and protein gene product [PGP] 9.5) with published markers (cytokeratins, glial fibrillary acidic protein [GFAP], progesterone receptor, S100, and vimentin). Neoplastic cells were immunohistochemically positive for vimentin in 100% of the meningiomas; CD34, 94%; GLUT-1, 86%; E-cadherin, 81%; S100, 75%; laminin, 72%; claudin-1, 60%; PGP 9.5, 55%; progesterone receptor, 44%; pancytokeratins, 39%; cytokeratins 8/18, 17%, and GFAP in 9%. Ki67 index did not correlate well with mitotic index. Based on these results and those in the human literature, immunohistochemistry for vimentin, CD34, and E-cadherin is proposed to support a diagnosis of meningioma. Immunohistochemistry for claudin-1, albeit of only moderate to low sensitivity in canine and feline meningiomas, may help to distinguish meningioma from some mesenchymal neoplasms involving the brain and associated structures, such as schwannomas, which in humans express claudin-1 poorly or not at all. Further studies with CD34, E-cadherin, and claudin-1 in canine and feline tumors that may mimic meningiomas are needed to determine the adequacy of this approach.