Immunohistochemical analyses of neural stem cell lineage markers in normal feline brains and glial tumors.

Neural stem cell (NSC) lineage cells have not been fully identified in feline brains, and the NSC-like nature of feline glial tumors has not been determined. In this study, 6 normal cat brains (3 newborn and 3 older cats) and 13 feline glial tumors were analyzed using immunohistochemical NSC lineage markers. The feline glial tumors were subjected to immunohistochemical scoring followed by hierarchical cluster analysis. In newborn brains, glial acidic fibrillary protein (GFAP)/nestin/sex-determining region Y-box transcription factor 2 (SOX2)-immunopositive NSCs, SOX2-immunopositive intermediate progenitor cells, oligodendrocyte transcription factor 2 (OLIG2)/platelet-derived growth factor receptor-α (PDGFR-α)-immunopositive oligodendrocyte precursor cells (OPCs), OLIG2/GFAP-immunopositive immature astrocytes, and neuronal nuclear (NeuN)/ß-3 tubulin-immunopositive mature neuronal cells were observed. The apical membrane of NSCs was also immunopositive for Na+/H+ exchanger regulatory factor 1 (NHERF1). In mature brains, the NSC lineage cells were similar to those of the newborn brains. A total of 13 glial tumors consisted of 2 oligodendrogliomas, 4 astrocytomas, 3 subependymomas, and 4 ependymomas. Astrocytomas, subependymomas, and ependymomas were immunopositive for GFAP, nestin, and SOX2. Subependymomas and ependymomas showed dot-like or apical membrane immunolabeling for NHERF1, respectively. Astrocytomas were immunopositive for OLIG2. Oligodendrogliomas and subependymomas were immunopositive for OLIG2 and PDGFR-α. Feline glial tumors also showed variable immunolabeling for ß-3 tubulin, NeuN, and synaptophysin. Based on these results, feline astrocytomas, subependymomas, and ependymomas appear to have an NSC-like immunophenotype. In addition, astrocytomas, subependymomas, and ependymomas have the characteristics of glial, oligodendrocyte precursor, and ependymal cells, respectively. Feline oligodendrogliomas likely have an OPC-like immunophenotype. In addition, feline glial tumors may have multipotential stemness for differentiation into neuronal cells. These preliminary results should be validated by gene expression analyses in future studies with larger case numbers.

Clinical impact of near-infrared fluorescence imaging with indocyanine green on surgical treatment for hepatic masses in dogs.

BACKGROUND: Near-infrared fluorescence imaging using indocyanine green (ICG) is clinically applied to intraoperatively identify hepatic masses in humans. In addition, it is reported to be effective for assessing complete resection in human hepatocellular carcinoma (HCC). However, there is limited information on ICG fluorescence imaging for canine HCC, and its clinical usefulness is still unclear. Therefore, the purpose of this study was to evaluate the intraoperative identification and status of surgical margin for canine hepatic masses using near-infrared ICG fluorescence imaging. This clinical study included 104 dogs with hepatic masses. Between 12 and 24 h prior to surgery, ICG solution was injected intravenously at a dose of 0.5 mg/kg. The fluorescence intensity and pattern of each hepatic mass was investigated using an infrared camera before resection. After resection, the fluorescence intensity of the resection margin was also investigated. The resected masses were histopathologically diagnosed and compared using ICG fluorescence imaging. RESULTS: One hundred and twenty-two masses obtained from 104 dogs included 76 HCCs, 16 hepatocellular adenomas, 12 focal nodular hyperplasias, and 18 other lesions. Of the 122 masses, 106 (94 partial, 9 whole, and 3 ring fluorescence patterns), 7, and 9 masses showed increased, the same, or decreased fluorescence compared to the normal liver tissue, respectively. The fluorescence intensity and pattern were not significantly related to the histopathological diagnosis. The sensitivity and specificity of the margin evaluation in the 47 dogs were 100% and 77.3%, respectively. The median survival times in cases of HCC with complete and incomplete resection were 914 and 254 days, respectively. The median survival time of patients with a complete resection was significantly longer than that of patients with a incomplete resection (p = 0.043). CONCLUSION: ICG fluorescence imaging has potential clinical value for the identification and margin evaluation of canine hepatic masses. Although it is difficult to use fluorescence imaging for the differential diagnosis of liver tumours, it may be useful for assessing complete resection in cases of hepatic masses demonstrating increased fluorescence in dogs, and complete resection of HCC could have a survival benefit.

Gastrointestinal stromal tumors with Kit gene mutation in 4 guinea pigs (Cavia porcellus).

Gastrointestinal stromal tumors (GISTs) have been rarely reported in guinea pigs. We aimed to characterize the clinical and pathological features of GISTs in 4 guinea pigs and investigate the presence of mutations in exon 11 of the KIT proto-oncogene receptor tyrosine kinase (Kit) gene. Two subjects were male and 2 were female; 2 were 6 years old, 1 was 7 years old, and 1 was of an unknown age. Three cases had primary gastric tumors, whereas 1 had a primary small intestinal tumor. All cases had tumors that extended from the submucosa to the serosa with extraluminal growth. A gastric tumor had gastric, pancreatic, and cecal metastases. Histologically, the tumors were sharply demarcated and composed of spindle cells arranged in bundles, intermixed with small amounts of collagenous stroma. The tumor cells had mild atypia with few mitotic figures (0-5/50 high power fields, 7.95 mm2) and were immunolabeled for KIT and Discovered-on-GIST 1 (DOG1). All cases had mutations in exon 11 of the Kit gene. These findings indicate that GISTs in guinea pigs are similar to those in humans and dogs. GISTs in guinea pigs are potentially malignant submucosal tumors with KIT- and DOG1-immunolabeling, exon 11 KIT mutations, and the possibility of metastasis.

Radiographic characteristics of canine subungual keratoacanthoma.

Subungual keratoacanthoma (SKA) is a rare benign nail bed tumor in dogs, and its radiographic characteristics have not been reported based on the authors' review of the literature. The purpose of this multicenter, retrospective, observational, descriptive study was to describe the radiographic features of SKA in dogs. Twelve dogs for a total of 12 digits with histologically confirmed SKA met the inclusion criteria. The radiographs of the manus or pes were reviewed by two veterinary radiologists and one veterinarian. The radiology reports were interpreted based on a consensus. In six dogs, there was lysis of both the middle phalanx (P2) and the distal phalanx (P3), whereas in the other six dogs, there was only lysis of P3. In all dogs with osteolysis of P2, the lysis involved the distal articular surface. Osteolysis of P3 was more severe in the ungual process than in the ungual crest in all dogs. The margins of the lytic regions of P2 and P3 were well defined and smoothly marginated in most dogs. Expansile changes in the P3 crest were observed in 83.3% (10/12 dogs), and the nail of the affected digit was enlarged and deformed in 91.6% (11/12 dogs). In summary, the radiographic features of canine SKA include severe pressure resorption of the P3 ungual process, expansile change of the P3 ungual crest, and nail enlargement and deformation. With these radiographic features, SKA should be considered as a differential diagnosis.

Long-term follow-up of portal vein thrombosis in an American Cocker Spaniel with lobular dissecting hepatitis: a case report.

BACKGROUND: Lobular dissecting hepatitis (LDH) is a rare form of canine liver cirrhosis that may be accompanied by portal hypertension in American Cocker Spaniels. In human patients with liver cirrhosis, portal vein thrombosis (PVT) is a common complication. However, PVT has not been reported in dogs with LDH. Herein, we describe the long-term follow-up of PVT in an American Cocker Spaniel with LDH. CASE PRESENTATION: An 8-year-old neutered male American Cocker Spaniel presented with a 1-month history of severe abdominal effusion. The dog was histopathologically diagnosed with LDH and treated with low-dose prednisolone on day 14. On day 115, computed tomography angiography (CTA) confirmed the presence of a thrombus in the portal vein. Therefore, the dog was subcutaneously administered with the anticoagulant dalteparin, and low-dose prednisolone was continued. As a follow-up for PVT, CTA examinations were performed on days 207, 515, 886, and 1168, and the dog's antithrombin and D-dimer levels were measured. Following anticoagulant therapy, the dog was confirmed to have gradually increased antithrombin activity and decreased D-dimer concentrations. In addition, although the thrombus was confirmed to be in the same area of the portal vein system by CTA, atrophy and increased CT values due to organization were observed during the follow-up period. The dog's condition remained stable without clinical signs until day 1112 when it developed hepatic encephalopathy. The dog died on day 1208. On postmortem examination, histopathologically, the liver showed marked bile duct hyperplasia and fibrosis with chronic thrombus in the portal vein. CONCLUSIONS: This case demonstrated that low-dose glucocorticoid combined with dalteparin allowed long-term follow-up of PVT in an American Cocker Spaniel with LDH.

International Guidelines for Veterinary Tumor Pathology: A Call to Action.

Standardization of tumor assessment lays the foundation for validation of grading systems, permits reproducibility of oncologic studies among investigators, and increases confidence in the significance of study results. Currently, there is minimal methodological standardization for assessing tumors in veterinary medicine, with few attempts to validate published protocols and grading schemes. The current article attempts to address these shortcomings by providing standard guidelines for tumor assessment parameters and protocols for evaluating specific tumor types. More detailed information is available in the Supplemental Files, the intention of which is 2-fold: publication as part of this commentary, but more importantly, these will be available as "living documents" on a website (www.vetcancerprotocols.org), which will be updated as new information is presented in the peer-reviewed literature. Our hope is that veterinary pathologists will agree that this initiative is needed, and will contribute to and utilize this information for routine diagnostic work and oncologic studies. Journal editors and reviewers can utilize checklists to ensure publications include sufficient detail and standardized methods of tumor assessment. To maintain the relevance of the guidelines and protocols, it is critical that the information is periodically updated and revised as new studies are published and validated with the intent of providing a repository of this information. Our hope is that this initiative (a continuation of efforts published in this journal in 2011) will facilitate collaboration and reproducibility between pathologists and institutions, increase case numbers, and strengthen clinical research findings, thus ensuring continued progress in veterinary oncologic pathology and improving patient care.

Prostaglandin E2 Induction Suppresses the Th1 Immune Responses in Cattle with Johne's Disease.

Johne's disease, caused by Mycobacterium avium subsp. paratuberculosis, is a bovine chronic infection that is endemic in Japan and many other countries. The expression of immunoinhibitory molecules is upregulated in cattle with Johne's disease, but the mechanism of immunosuppression is poorly understood. Prostaglandin E2 (PGE2) is immunosuppressive in humans, but few veterinary data are available. In this study, functional and kinetic analyses of PGE2 were performed to investigate the immunosuppressive effect of PGE2 during Johne's disease. In vitro PGE2 treatment decreased T-cell proliferation and Th1 cytokine production and upregulated the expression of immunoinhibitory molecules such as interleukin-10 and programmed death ligand 1 (PD-L1) in peripheral blood mononuclear cells (PBMCs) from healthy cattle. PGE2 was upregulated in sera and intestinal lesions of cattle with Johne's disease. In vitro stimulation with Johnin purified protein derivative (J-PPD) induced cyclooxygenase-2 (COX-2) transcription, PGE2 production, and upregulation of PD-L1 and immunoinhibitory receptors in PBMCs from cattle infected with M. avium subsp. paratuberculosis Therefore, Johnin-specific Th1 responses could be limited by the PGE2 pathway in cattle. In contrast, downregulation of PGE2 with a COX-2 inhibitor promoted J-PPD-stimulated CD8+ T-cell proliferation and Th1 cytokine production in PBMCs from the experimentally infected cattle. PD-L1 blockade induced J-PPD-stimulated CD8+ T-cell proliferation and interferon gamma production in vitro Combined treatment with a COX-2 inhibitor and anti-PD-L1 antibodies enhanced J-PPD-stimulated CD8+ T-cell proliferation in vitro, suggesting that the blockade of both pathways is a potential therapeutic strategy to control Johne's disease. The effects of COX-2 inhibition warrant further study as a novel treatment of Johne's disease.

New World feline APOBEC3 potently controls inter-genus lentiviral transmission.

BACKGROUND: The apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3; A3) gene family appears only in mammalian genomes. Some A3 proteins can be incorporated into progeny virions and inhibit lentiviral replication. In turn, the lentiviral viral infectivity factor (Vif) counteracts the A3-mediated antiviral effect by degrading A3 proteins. Recent investigations have suggested that lentiviral vif genes evolved to combat mammalian APOBEC3 proteins, and have further proposed that the Vif-A3 interaction may help determine the co-evolutionary history of cross-species lentiviral transmission in mammals. RESULTS: Here we address the co-evolutionary relationship between two New World felids, the puma (Puma concolor) and the bobcat (Lynx rufus), and their lentiviruses, which are designated puma lentiviruses (PLVs). We demonstrate that PLV-A Vif counteracts the antiviral action of APOBEC3Z3 (A3Z3) of both puma and bobcat, whereas PLV-B Vif counteracts only puma A3Z3. The species specificity of PLV-B Vif is irrespective of the phylogenic relationships of feline species in the genera Puma, Lynx and Acinonyx. We reveal that the amino acid at position 178 in the puma and bobcat A3Z3 is exposed on the protein surface and determines the sensitivity to PLV-B Vif-mediated degradation. Moreover, although both the puma and bobcat A3Z3 genes are polymorphic, their sensitivity/resistance to PLV Vif-mediated degradation is conserved. CONCLUSIONS: To the best of our knowledge, this is the first study suggesting that the host A3 protein potently controls inter-genus lentiviral transmission. Our findings provide the first evidence suggesting that the co-evolutionary arms race between lentiviruses and mammals has occurred in the New World.

Expression of Oligodendrocyte Precursor Cell Markers in Canine Oligodendrogliomas.

Oligodendroglioma is a common brain tumor in dogs, particularly brachycephalic breeds. Oligodendrocyte precursor cells (OPCs) are suspected to be a possible origin of oligodendroglioma, although it has not been well elucidated. In the present study, 27 cases of canine brain oligodendrogliomas were histologically and immunohistochemically examined. The most commonly affected breed was the French Bulldog ( n = 19 of 27, 70%). Seizure was the most predominant clinical sign ( n = 17 of 25, 68%). The tumors were located mainly in the cerebrum, particularly in the frontal lobe ( n = 10 of 27, 37%). All cases were diagnosed as anaplastic oligodendroglioma (AO) and had common histologic features characterized by the proliferation of round to polygonal cells with pronounced atypia and conspicuous mitotic activity (average, 10.7 mitoses per 10 high-power fields). Honeycomb pattern ( n = 5 of 27, 19%), myxoid matrix ( n = 10, 37%), cyst formation ( n = 6, 22%), necrosis ( n = 19, 70%), pseudopalisading ( n = 5, 18.5%), glomeruloid vessels ( n = 16, 59%), and microcalcification ( n = 5, 19%) were other histopathologic features of the present tumors. Immunohistochemically, the tumor cells were positive for Olig2 in all cases and for other markers of OPCs in most cases, including SOX10 ( n = 24 of 27, 89%), platelet-derived growth factor receptor α ( n = 24, 89%), and NG2 ( n = 23, 85%). The present AO also consisted of heterogeneous cell populations that were positive for nestin ( n = 13 of 27, 48%), glial fibrillary acidic protein ( n = 5, 19%), doublecortin ( n = 22, 82%), and ßIII-tubulin ( n = 15, 56%). Moreover, cultured AO cells obtained from 1 case retained expression of OPC markers and exhibited multipotent characteristics in a serum culture condition. Overall, the findings suggest that transformed multipotent OPCs may be a potential origin of canine AO.

Diastolic heart failure associated with hemangiosarcoma infiltrating left ventricular walls in a dog.

A 9-year-old Shetland sheepdog was diagnosed with cardiogenic pulmonary edema. Echocardiography revealed focally thickened left ventricular free wall and interventricular septum and left atrial dilation. Left ventricular systolic function was preserved. Doppler echocardiography of transmitral flow indicated restrictive left ventricular filling. Cardiac histopathology demonstrated hemangiosarcoma infiltrating the left ventricular walls.

Insuffisance cardiaque diastolique associée à l'infiltration d'un hémangiosarcome dans les parois ventriculaires gauches chez un chien. Un chien berger des Shetlands âgé de 9 ans a été diagnostiqué avec un oedème pulmonaire cardiogénique. L'échocardiographie a révélé un épaississement focal de la paroi libre du ventricule gauche et du septum interventriculaire et une dilatation atriale gauche.. La fonction systolique ventriculaire gauche était préservée. L'échographie Doppler de l'écoulement transmitral indiquait un remplissage ventriculaire gauche restreint. L'histopathologie cardiaque a démontré l'infiltration des parois ventriculaires gauches par l'hémangiosarcome.(Traduit par Isabelle Vallières).

Bovine Immunoinhibitory Receptors Contribute to Suppression of Mycobacterium avium subsp. paratuberculosis-Specific T-Cell Responses.

Johne's disease (paratuberculosis) is a chronic enteritis in cattle that is caused by intracellular infection with Mycobacterium avium subsp. paratuberculosis. This infection is characterized by the functional exhaustion of T-cell responses to M. avium subsp. paratuberculosis antigens during late subclinical and clinical stages, presumably facilitating the persistence of this bacterium and the formation of clinical lesions. However, the mechanisms underlying T-cell exhaustion in Johne's disease are poorly understood. Thus, we performed expression and functional analyses of the immunoinhibitory molecules programmed death-1 (PD-1)/PD-ligand 1 (PD-L1) and lymphocyte activation gene 3 (LAG-3)/major histocompatibility complex class II (MHC-II) in M. avium subsp. paratuberculosis-infected cattle during the late subclinical stage. Flow cytometric analyses revealed the upregulation of PD-1 and LAG-3 in T cells in infected animals, which suffered progressive suppression of interferon gamma (IFN-γ) responses to the M. avium subsp. paratuberculosis antigen. In addition, PD-L1 and MHC-II were expressed on macrophages from infected animals, consistent with PD-1 and LAG-3 pathways contributing to the suppression of IFN-γ responses during the subclinical stages of M. avium subsp. paratuberculosis infection. Furthermore, dual blockade of PD-L1 and LAG-3 enhanced M. avium subsp. paratuberculosis-specific IFN-γ responses in blood from infected animals, and in vitro LAG-3 blockade enhanced IFN-γ production from M. avium subsp. paratuberculosis-specific CD4(+) and CD8(+) T cells. Taken together, the present data indicate that M. avium subsp. paratuberculosis-specific T-cell exhaustion is in part mediated by PD-1/PD-L1 and LAG-3/MHC-II interactions and that LAG-3 is a molecular target for the control of M. avium subsp. paratuberculosis-specific T-cell responses.

Preoperative Differential Diagnosis of Canine Adrenal Tumors Using Triple-Phase Helical Computed Tomography.

OBJECTIVE: To characterize the computed tomography (CT) findings for canine adrenal tumors, including cortical adenoma, cortical adenocarcinoma, and pheochromocytoma, and to evaluate the feasibility and usefulness of preoperative triple-phase helical CT for differentiation of tumor types and surgical planning. STUDY DESIGN: Retrospective study. ANIMALS: Dogs with adrenal tumors (n=36). METHODS: All dogs underwent triple-phase helical CT, followed by adrenalectomy and histopathological diagnosis of the resected mass. Precontrast images, arterial, venous, and delayed phase images were obtained. In all cases, morphological characteristics and CT values and calculations, including the percentage enhancement washout ratio, relative percentage washout, enhancement washin, and enhancement washout, were analyzed and compared among the tumor types. RESULTS: Of the 36 dogs with adrenal masses, cortical adenocarcinoma was most commonly diagnosed (16 dogs), followed by pheochromocytoma (13 dogs), and cortical adenoma (7 dogs). The precontrast minimum CT value and enhancement washout between venous and delayed phases in the cortical adenoma were significantly higher than those in the cortical adenocarcinoma. The maximum CT values of the precontrast image and arterial and venous phases, the enhancement washin and washouts, percentage enhancement washout ratio, and relative percentage washout in the pheochromocytomas were significantly higher than those in cortical adenocarcinoma. CONCLUSION: The differential diagnosis of canine adrenal tumors was feasible based on triple-phase CT findings, including morphological features, CT values, and intratumoral contrast attenuation. Preoperative diagnosis using triple-phase helical CT may be useful for surgical planning in dogs with adrenal tumors.

Squamous cell carcinoma causing dorsal atlantoaxial spinal cord compression in a dog.

A 12-year-old Chihuahua dog was presented for cervical pain and progressive tetraparesis. Magnetic resonance imaging revealed spinal cord compression due to a mass in the dorsal atlantoaxial region. Surgical treatment was performed. The mass was histopathologically diagnosed as a squamous cell carcinoma. The dog recovered to normal neurologic status after surgery.

Carcinome squameux causant une compression de la moelle épinière altantoaxiale dorsale chez un chien. Un Chihuahua âgé de 12 ans a été présenté pour de la douleur cervicale et une tétraparésie progressive. Une imagerie par résonance magnétique a révélé une compression de la moelle épinière en raison d'une masse dans la région atlantoaxiale dorsale. Le traitement chirurgical a été réalisé. La masse a été diagnostiquée à l'histopathologie comme étant un carcinome squameux. Le chien a retrouvé une condition neurologique normale après l'opération.(Traduit par Isabelle Vallières).

Triple-phase helical computed tomography in dogs with hepatic masses.

The purpose of this study was to determine the utility of triple-phase helical computed tomography (CT) for differentiating canine hepatic masses. Seventy dogs with hepatic masses underwent triple-phase CT followed by surgical removal of the hepatic masses. Triple-phase helical CT scans for each dog included precontrast, arterial phase, portal venous phase, and delayed phase studies. The removed hepatic masses were histopathologically classified as hepatocellular carcinoma (n = 47), nodular hyperplasia (n = 14), and hepatic metastatic tumors (n = 9) in dogs. Of the 47 hepatocellular carcinomas, the most common CT findings included a heterogeneous pattern with hyper-, iso-, and hypoenhancement in both the arterial and portal venous phases (40/47, 85.1%). Of the 14 nodular hyperplasias, the most common CT findings were a homogeneous pattern with hyper- and isoenhancement in both the portal venous and delayed phases (13/14, 92.9%). Of nine hepatic metastatic tumors, the most common CT findings included a homogeneous hypoenhancement pattern in both the arterial and portal venous phases (8/9, 88.9%). In addition, 5 (55.6%) showed homogeneous hypoenhancement patterns in the delayed phase. Findings from our study indicated that triple-phase CT is a useful tool for preoperative differentiation of hepatocellular carcinoma, nodular hyperplasia, and hepatic metastatic tumors in dogs.

Jaundice caused by primary common bile duct lymphoma of a cat.

A 10-year-old American Shorthair cat presented with anorexia and jaundice, and echogenic evaluation revealed diffuse thickening of the common bile duct (CBD) wall. An exploratory laparotomy was conducted, the lesion was evaluated as difficult to remove, and the cat was euthanized and autopsied. Histologically, round neoplastic cells proliferated in the mucosa of the CBD and infiltrated the hepatic lobe, pancreas, and duodenum. Immunohistochemistry revealed that the neoplastic cells were positive for cytoplasmic-CD3 and granzyme B, and TCR-gamma clonal rearrangement was detected. Based on these findings, the neoplasia was diagnosed as a primary CBD lymphoma originating from cytotoxic T or natural killer cells. To the best of our knowledge, this is the first reported case of feline primary CBD lymphoma. Although rare, lymphoma of the CBD should be considered in cats with jaundice and thickening of the CBD.

Improved detection of air-filled lesions using computed tomography in dogs with recurrent spontaneous pneumothorax through reduction of pulmonary atelectasis via positive pressure ventilation.

Introduction: Spontaneous pneumothorax in dogs is predominantly caused by the rupture of air-filled lesions, such as bullae or blebs. The efficacy of Computed Tomography (CT) in detecting these lesions has been deemed limited due to its reportedly low sensitivity. This retrospective, cross-sectional study investigates the utility of CT in eight dogs diagnosed with recurrent pneumothorax, all of which had surgical confirmation of the cause of the pneumothorax. Materials and methods: Thoracic radiographs were obtained before and the day following the CT studies. Initially, a CT study was conducted without positive pressure ventilation (pre-PPV CT). Subsequent CT studies were performed post-evacuation of pneumothorax and with positive pressure ventilation of 15 cmH2O until lung atelectasis was resolved (post-PPV CT). The pre-PPV CT and post-PPV CT images were anonymized and reviewed by two board-certified radiologists. The presence and morphology of air-filled lesions were evaluated on all images. Surgical findings were recorded and compared to the CT findings. Results: Air-filled lesions were detected in 5 out of 8 dogs in the pre-PPV CT studies and in all 8 dogs in the post-PPV CT studies. The CT findings of air-filled lesions were consistent with surgical findings. None of the dogs showed increased severity of pneumothorax in radiographs taken the day following the CT studies. Discussions: The study concludes that the resolution of lung atelectasis by evacuation of pneumothorax and positive pressure ventilation during CT studies is feasible and enhances the detection of air-filled lesions in dogs with recurrent spontaneous pneumothorax. This could potentially aid in improving surgical planning.

Use of genome-wide DNA methylation analysis to identify prognostic CpG site markers associated with longer survival time in dogs with multicentric high-grade B-cell lymphoma.

BACKGROUND: DNA methylation analysis might identify prognostic CpG sites in CHOP-treated dogs with multicentric high-grade B-cell lymphoma (MHGL) with heterogenous prognosis. OBJECTIVE: To identify prognostic CpG sites of MHGL through genome-wide DNA methylation analysis with pyrosequencing validation. ANIMALS: Test group: 24 dogs. Validation group: 100 dogs. All client-owned dogs were diagnosed with MHGL and treated with CHOP chemotherapy. METHODS: Cohort study. DNA was extracted from lymph node samples obtained via FNA. Genome-wide DNA methylation analysis using Digital Restriction Enzyme Analysis of Methylation (DREAM) was performed on the test group to identify differentially methylated CpG sites (DMCs). Bisulfite pyrosequencing was used to measure methylation status of candidate DMCs in the validation group. Median survival times (MST) were analyzed using Kaplan-Meier (log-rank) product limit method. RESULTS: DREAM analyzed 101 576 CpG sites. Hierarchical clustering of 16 262 CpG sites in test group identified group with better prognosis (MST = 55-477 days vs 10-301 days, P = .007). Volcano plot identified 1371 differentially methylated CpG sites (DMCs). DMC near the genes of FAM213A (DMC-F) and PHLPP1 (DMC-P) were selected as candidates. Bisulfite-pyrosequencing performed on validation group showed group with methylation level of DMC-F < 40% had favorable prognosis (MST = 11-1072 days vs 8-1792 days, P = .01), whereas group with the methylation level combination of DMC-F < 40% plus DMC-P < 10% had excellent prognosis (MST = 18-1072 days vs 8-1792 days, P = .009). CONCLUSION AND CLINICAL IMPORTANCE: Methylation status of prognostic CpG sites delineate canine MGHL cases with longer MST, providing owners with information on expectations of potential improved treatment outcomes.

A case of immature teratoma with nephroblastic components in the oral cavity of a cat.

A 2-month-old female mixed cat was emaciated due to dysphagia, and inspection of the mouth revealed a 2 cm pedunculated mass elongated from the palate, which occupied the oral cavity. The mass was surgically removed, and histopathological examination revealed that the tumor was composed of three germ cell layers: ectodermal (skin and skin appendages), mesodermal (cartilaginous and osseous structures), and endodermal (glandular and respiratory mucosa) tissues. An immature teratoma was diagnosed because of the presence of immature neuroectodermal tissues, and the presence of nephroblastic components was a characteristic finding in this case. This is the first report of an oropharyngeal teratoma in cats and the first case of an immature teratoma with nephroblastic components in a domestic species.

Impact of near-infrared fluorescence imaging with indocyanine green on the surgical treatment of pulmonary masses in dogs.

Objectives: To investigate the intraoperative identification and complete resection of pulmonary masses, and to evaluate lymph node metastasis of pulmonary malignant tumors in dogs using indocyanine green (ICG) fluorescence imaging. Methods: Forty dogs with pulmonary masses were included, all of which underwent surgical treatment. ICG fluorescence imaging was performed on pulmonary masses before lobectomy and the resection margins after lobectomy. In addition, ICG fluorescence of the excised masses and lymph nodes was evaluated in the shaded box. The fluorescence findings were compared with the histopathological diagnosis. Results: Of 44 nodules resected from 40 dogs, 32 nodules were histopathologically diagnosed as lung adenocarcinoma, five were histiocytic sarcoma, three were undifferentiated sarcoma, two were malignant epithelial tumor metastases, one was carcinosarcoma, and one was a non-neoplastic lesion. Fluorescence was observed in all nodules. In addition to the main lesion, other fluorescent nodules were found in four dogs. Regarding the diagnostic accuracy of complete resection based on ICG fluorescence, the sensitivity was 67.7% and the specificity was 60.0%. The sensitivity and specificity of ICG fluorescence for the diagnosis of lymph node metastasis were 100 and 75.0%, respectively. Conclusions: ICG fluorescence imaging might be a useful intraoperative diagnostic method to identify the location of tumors and lymph node metastasis, but not to evaluate complete tumor resection, in dogs with pulmonary malignant tumors.

Cross-reactivity of anti-human programmed cell death ligand 1 (PD-L1) monoclonal antibody, clone 28-8 against feline PD-L1.

Immunotherapy is a breakthrough in human cancer therapy and has become a major concern in veterinary oncology. However, in cats, many unclear points of the tumor microenvironment exist, including immune checkpoint molecules. A reason is that very few monoclonal antibodies have been proven to react with feline molecules. Therefore, this study investigated whether anti-human programmed cell death ligand 1 (PD-L1) monoclonal antibody, clone 28-8, which is currently commercially available, can also recognize feline PD-L1 by flow cytometry, immunoprecipitation, and immunohistochemical (IHC) staining. We confirmed that the antibody's specificity by flow cytometry and immunoprecipitation using NIH3T3 cells transfected with feline PD-L1. Additionally, we revealed that PD-L1 was expressed on the surface of some feline cell lines by flow cytometry and clone 28-8 antibody unbound to the cells where feline PD-L1 was knocked out. Furthermore, IHC analysis revealed that PD-L1 was expressed in macrophages in the spleen and lymph nodes from healthy cats and mast cell tumor cells. Therefore, we indicated that the clone 28-8 antibody is a valuable tool in detecting feline PD-L1, and further analysis of tumor tissues is expected in the future.