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Abstract Background Severe muscle stiffness (SMS) in dogs with hypercortisolism (HC) is uncommon. Objectives To evaluate signalment, presentation, treatments, and long‐term outcomes of dogs with concurrent HC and SMS. Animals Thirty‐seven dogs. Methods Medical records of dogs with HC and concurrent SMS were recruited from 10 institutions. Clinical information, test results, therapeutic responses, and survival times were reviewed. Results All 37 dogs with HC and SMS had pituitary‐dependent hypercortisolism (PDH); 36/37 weighed

Abstract Erythroleukaemia is a malignant neoplasm of the erythroid lineage that rarely occurs in cats. It is associated with the feline leukaemia virus (FeLV), and owing to the poor prognosis, treatment is rarely reported. A 4‐year‐old female Korean domestic shorthair cat was presented with hyporexia, fever, lethargy, severe anaemia and rubricytosis. An FeLV antigen test was positive, but a subsequent polymerase chain reaction test was negative. Serum biochemistry analysis results were normal, except for slightly elevated alanine aminotransferase. The patient was tentatively diagnosed with acute erythroleukaemia, and single high‐dose (600 mg/m2) cytarabine chemotherapy was administered via constant rate infusion for 12 h a day for 5 days. After the first cytarabine administration, the clinical signs and anaemia improved, though no change was noted to other haematological parameters. The patient died of shock 16 days after the second cytarabine administration; the total survival time after diagnosis was 67 days. Post‐mortem cytological evaluation of bone marrow aspiration revealed that the myeloid/erythroid ratio was 0.49, the erythroid progenitor cells were 64% of all nucleated cells and the blast cells were 84% of the non‐erythroid cells. Histopathology images indicated that the spleen was diffusely expanded by atypical round cells, possibly erythroid precursors. This is the first case report on the prognosis and effects of high‐dose cytarabine chemotherapy for acute feline erythroleukaemia with FeLV infection. Although the clinical signs improved, the treatment was not effective. Further studies on erythroleukaemia chemotherapy protocols are required.

Abstract Lymphoma is widely recognized in veterinary medicine. However, studies focused on secondary lymphoma after chemotherapy do not exist in veterinary medicine. An 11‐year‐old, spayed female Shih‐Tzu dog was diagnosed with mammary gland carcinoma. Twenty‐five months after carboplatin treatment, the dog developed generalized lymphadenopathy (GL), diagnosed as high‐grade T‐cell lymphoma by immunohistochemistry. Another spayed female Shih‐Tzu dog who was 15‐year‐old had biopsy‐induced gastrointestinal stromal tumour. Three months after being treated with Toceranib, the dog developed GL that was diagnosed by PCR for antigen receptor rearrangement as T‐cell lymphoma. An eight‐year‐old, castrated male Mongrel dog was diagnosed with mast cell tumour. The dog was treated with vinblastine, but 14 months later, GL was revealed. Fine‐needle aspiration indicated lymphoma. The owner declined to investigate the cell lineage. All three dogs developed GL after chemotherapy. We suggest that secondary lymphoma can develop in dogs after chemotherapy for a primary cancer, and thus long‐term follow‐up is necessary.

Abstract Background Stress is associated with various detrimental changes in physiological health that affect an animal’s quality of life. The hypothalamus-pituitary-adrenal (HPA) axis and the sympathetic-adreno-medullar (SAM) axis are two main physiological pathways that constitute the stress response of an organism. Arginine vasopressin (AVP) is a mediator of the HPA axis and is known to be related to social behaviours and stress. The serum concentration of AVP is higher in more aggressive dogs and humans with post-traumatic stress disorder. Salivary biomarker analysis is a non-invasive method to assess stress. The purpose of this study was to evaluate the possibility of using salivary AVP as an acute stress biomarker in dogs. Salivary AVP concentration was measured before and after exposure to all relevant environmental stimuli (i.e. car trip to the lab, physical examination by the veterinarian, and sampling procedure,) and then after 30 min of vacuum noise exposure. Behavioural assessments, physiologic parameter assessments, and serum cortisol analysis were conducted in combination. Statistical analysis was conducted separately in the total study population, the less stressed group, and the more stressed group, respectively. Results Based on stress behaviour analysis scores, 28 dogs were classified into less or more stressed groups. All four physiologic parameters (blood pressure, body temperature, heart rate, and respiratory rate) were significantly increased after noise and environmental challenges, in the more stressed group. Serum cortisol did not show any significant change. Salivary AVP significantly decreased after noise and environmental stimulation in the more stressed group but not in the less stressed group. Salivary AVP and blood pressure changes were negatively correlated in the more stressed group. Conclusion Salivary AVP may be a potential acute stress biomarker in dogs.

Abstract A 12‐year‐old female Shih‐tzu dog was presented with a 2‐month history of cutaneous non‐pruritic multiple ulcerated or crusted nodules of less than 1.5 cm in diameter on eyelids, lips, abdomen, groin, thighs and perianal region. Several diagnostic tests were performed, including fine needle aspiration and skin biopsy of the cutaneous nodules. Cytologic interpretation was round cell neoplasm with multilobated (clover‐leaf shaped) nuclei. Histopathology revealed round neoplastic cells with prominent anisocytosis and anisokaryosis, and numerous mitotic figures; however, the origin of the cells was not identified. Immunohistochemical evaluation indicated that these cells were positive for CD79a and MUM‐1, but negative for CD3, CD20 and Pax 5. The patient was treated with chemotherapy, and the skin condition improved. Despite good response to chemotherapy, the patient was euthanized due to poor general health.

Abstract Background As a co-receptor for fibroblast growth factor 23, klotho plays a pivotal role in phosphate metabolism. The kidney is known to be the main source of soluble alpha-klotho and the principal regulator of its concentration. Previous studies in human participants showed that the concentration of soluble alpha-klotho in serum and urine decreased in chronic kidney disease (CKD) patients. However, no previous study has assessed soluble alpha-klotho levels in dogs. This study aimed to measure serum and urinary alpha-klotho levels in CKD dogs and identify their associations with International Renal Interest Society (IRIS) CKD stages and other parameters known to be associated with CKD. Results Serum and urinary alpha klotho concentrations were measured by a commercially available canine-specific sandwich enzyme-linked immunosorbent assay kit and compared between groups by a nonparametric Kruskal–Wallis test. Spearman’s correlation coefficient was used to evaluate the relationships between variables. A stepwise multiple regression analysis was performed to estimate the effects of independent predictors on klotho concentrations. The urine klotho-to-creatinine ratio (UrKl/Cr) was significantly lower in stage 3 dogs than the control group and was significantly lower in dogs with stage 3 and 4 CKD than in those with stage 1 and 2 disease. UrKl/Cr was negatively correlated with serum symmetric dimethylarginine (sSDMA), blood urea nitrogen (BUN), creatinine, and phosphorus concentration. Serum alpha-klotho concentration in dogs with stages 2 and 3 CKD was significantly lower than those in the control group. There was no significant correlation between serum alpha-klotho and BUN, creatinine, and phosphorus concentrations. No statistically significant differences were observed in UrKl/Cr and serum alpha-klotho concentration between groups based on sex, age, urine protein-to-creatinine ratio (UPC), or blood pressure. Conclusions UrKl/Cr decreased in dogs with advanced CKD, and it was negatively correlated with sSDMA, BUN, creatinine, and phosphorus concentrations. Thus, klotho is associated with CKD and its clinical consequences, including CKD-mineral bone disorder, in dogs. Although serum klotho concentration was negatively correlated with sSDMA levels, it was not apparently related to IRIS CKD stage or other parameters known to be associated with CKD.

Adipose tissue-derived mesenchymal stem cells (AT-MSCs) are attractive cell-therapy vehicles for the delivery of anti-tumor molecules into the tumor microenvironment. The innate tropism of AT-MSCs for tumors has important implications for effective cellular delivery of anti-tumor molecules, including cytokines, interferon, and pro-drugs. The present study was designed to determine the possibility that the combination of stem cell-based gene therapy with low-dose cisplatin would improve therapeutic efficacy against canine melanoma. The IFN-β transduced canine AT-MSCs (cAT-MSC-IFN-β) inhibited the growth of LMeC canine melanoma cells in direct and indirect in vitro co-culture systems. In animal experiments using BALB/c nude mouse xenografts, which developed by injecting LMeC cells, the combination treatment of cAT-MSC-IFN-β and low-dose cisplatin significantly reduced tumor volume compared with the other treatment groups. Fluorescent microscopic analysis with a TUNEL (terminal deoxynucleotidyl transferase-mediated nick-end labeling) assay of tumor section provided evidence for homing of cAT-MSC-IFN-β to the tumor site and revealed that the combination treatment of cAT-MSC-IFN-β with low-dose cisplatin induced high levels of cell apoptosis. These findings may prove useful in further explorations of the application of these combined approaches to the treatment of malignant melanoma and other tumors.

Despite significant advancements in osteosarcoma research, the overall survival of canine and human osteosarcoma patients has remained essentially static over the past 2 decades. Post-operative limb-spare infection has been associated with improved survival in both species, yet a mechanism for improved survival has not been clearly established. Given that the majority of canine osteosarcoma patients experiencing post-operative infections were treated with fluoroquinolone antibiotics, we hypothesized that fluoroquinolone antibiotics might directly inhibit the survival and proliferation of canine osteosarcoma cells. Ciprofloxacin or enrofloxacin were found to inhibit p21(WAF1) expression resulting in decreased proliferation and increased S-G(2)/M accumulation. Furthermore, fluoroquinolone exposure induced apoptosis of canine osteosarcoma cells as demonstrated by cleavage of caspase-3 and PARP, and activation of caspase-3/7. These results support further studies examining the potential impact of quinolones on survival and proliferation of osteosarcoma.

Abstract Background Endothelial cell-specific molecule-1 (ESM-1) has emerged as a potential biomarker for cardiovascular disease in humans. Myxomatous mitral valve disease (MMVD) is the most common heart disease in dogs, and we hypothesized that MMVD causes chronic inflammation that increases susceptibility to endothelial glycocalyx (eGCX) damage. In this study, we measured the concentration of ESM-1 in a group of dogs with MMVD and evaluated factors affecting eGCX damage. Results Sixty-four dogs (control, n = 6; MMVD, n = 58) were enrolled in this study. There was no significant difference in serum ESM-1 concentrations among the MMVD stages. The serum ESM-1 concentration was significantly higher in the death group than in the alive group in MMVD dogs. (p = 0.006). In five dogs with MMVD, serum ESM-1 concentrations tended to decrease when the cardiac drug (pimobendan, furosemide, and digoxin) dose was increased. Conclusions In cases where MMVD progressed to decompensated heart failure with clinical symptoms and resulted in death, the concentration of serum ESM-1 increased significantly. Therefore, ESM-1 could be utilized as a new potential negative prognostic factor in patients with MMVD.

Abstract Background The use of fenbendazole (FBZ) in terminal cancer patients has recently increased, as anthelminthic drugs, such as FBZ and benzimidazole, exhibit anti‐tubulin effects in tumour cells. Objectives The present study evaluated the in vitro anti‐cancer effects of FBZ in five canine melanoma cell lines originating from the oral cavity (UCDK9M3, UCDK9M4, UCDK9M5, KMeC and LMeC). Methods Five canine melanoma cell lines were treated with FBZ and analysed with cell viability assay, cell cycle analysis, western blot assay and immunofluorescence staining to identify apoptotic effect, cell cycle arrest, microtubule disruption and mitotic slippage. Results Cell viability was reduced in all melanoma cell lines in a dose‐dependent manner after FBZ treatment. Through cell cycle analysis, G2/M arrest and mitotic slippage were identified, which showed a time‐dependent change. All treatment concentrations induced increased cleaved PARP signals in western blot analysis compared to the control groups. Immunofluorescence of cells treated for 24 h revealed defects in microtubule structure, multinucleation or macronucleation. With the exception of UCDK9M3, the melanoma cells showed mitotic slippage and post‐slippage death, indicative of mitotic catastrophe. Conclusions These results indicate that FBZ exhibits anti‐cancer effects in vitro against canine melanoma cells; however, further in vivo studies regarding the clinical applications of FBZ are required.

Abstract Background Gallbladder mucocele (GBM) is a common biliary disorder in dogs that can be categorized into 6 types, but the value of this classification scheme remains unknown. Cholecystectomy is associated with high death rates and warrants additional interrogation. Objectives Investigate the clinical value of ultrasonographic diagnosis of type of GBM and identify prognostic factors in dogs with GBM undergoing cholecystectomy. Animals Two hundred sixteen dogs. Methods Retrospective cohort study. Dogs with GBM diagnosed from 2014 to 2019 at 6 veterinary referral hospitals in Asia. Ultrasonogram images were reviewed and a GBM type (ie, types I‐VI) assigned. Results Dogs with GBM type V as compared to I (OR, 8.6; 95% CI, 2.6‐27.8; P

Importance: This is the first reported case of fragrance products-induced recurrent oxidative hemolytic anemia in a dog, detailing the successful therapeutic approach employed. Case Presentation: A 4-year-old intact female Pomeranian dog presented with brown tongue, pigmenturia, peripheral edema, and vomiting. Blood smears revealed a high count of eccentrocytes and Heinz bodies, along with a precipitous decline in packed cell volume and an increase in blood methemoglobin levels, suggesting an oxidative hemolytic crisis. This clinicopathological pattern recurred several times after the patient returned home. Antioxidants, methylene blue, hyperbaric oxygen (HBO) therapy, and blood transfusion were successfully employed to address recurrent hemolytic anemia; however, oxidative hemolytic crises recurred. After the owner removed exposure to various home remedies and fragrances, the clinical signs and hemolytic crises did not recur. Conclusions and Relevance: Recurring oxidative hemolytic crises should raise suspicions of environmental toxicity, which, although harmless in small quantities to humans, can be devastating to small-breed dogs. In addition to removing the causative agents, methylene blue and other antioxidants, along with HBO, may be beneficial in the acute management of oxidative hemolytic anemia. [ABSTRACT FROM AUTHOR]

Background: The grade of tracheal collapse (TC) is assessed by the diameter of the narrowed lumen. However, studies on the relationship between TC grade and clinical symptom severity are lacking. Objectives: To investigate the clinical characteristics of small-breed dogs diagnosed with TC and determine if fluoroscopic grading correlates with cough severity. Methods: We retrospectively reviewed medical records from 2022 to 2024. TC diagnosis was confirmed using fluoroscopic examination. Multiple linear regression was employed to investigate factors influencing cough severity, with a significance level set at p < 0.05. Results: A total of 132 dogs with TC were identified, of which 22 were excluded. The final cohort consisted of 110 dogs, aged between 2-19 years, with no significant sex differences. The majority (97.2%) of dogs had a BCS of =4. Among the top four breeds (Maltese, Pomeranian, Poodle, and Chihuahua), the most severe collapse was observed in the carinal region. The grade of collapse on fluoroscopy was mostly related to high BCS (p < 0.007) and low body weight (p < 0.001). However, interestingly, fluoroscopic findings of collapse location and grade did not correlate with cough severity (p = 0.350). Notably, clinical symptoms improved in 86.6% of cases following interventions such as weight reduction, environmental changes, and pharmacotherapy. Conclusions and clinical relevance: In small-breed dogs, the severity of cough was not associated with the region or grade of TC diagnosed by fluoroscopy. [ABSTRACT FROM AUTHOR]

Background: Axitinib, a potent and selective inhibitor of vascular endothelial growth factor (VEGF) receptor (VEGFR) tyrosine kinase 1,2 and 3, is used in chemotherapy because it inhibits tumor angiogenesis by blocking the VEGF/VEGFR pathway. In veterinary medicine, attempts have been made to apply tyrosine kinase inhibitors with anti-angiogenic effects to tumor patients, but there are no studies on axitinib in canine mammary gland tumors (MGTs). Objectives: This study aimed to confirm the antitumor activity of axitinib in canine mammary gland cell lines. Methods: We treated canine MGT cell lines (CIPp and CIPm) with axitinib and conducted CCK, wound healing, apoptosis, and cell cycle assays. Additionally, we evaluated the expression levels of angiogenesis-associated factors, including VEGFs, PDGF-A, FGF-2, and TGF-β1, using quantitative real-time polymerase chain reaction. Furthermore, we collected canine peripheral blood mononuclear cells (PBMCs), activated them with concanavalin A (ConA) and lipopolysaccharide (LPS), and then treated them with axitinib to investigate changes in viability. Results: When axitinib was administered to CIPp and CIPm, cell viability significantly decreased at 24, 48, and 72 h ( p < 0.001), and migration was markedly reduced (6 h, p < 0.05; 12 h, p < 0.005). The apoptosis rate significantly increased ( p < 0.01), and the G2/M phase ratio showed a significant increase ( p < 0.001). Additionally, there was no significant change in the viability of canine PBMCs treated with LPS and ConA. Conclusion: In this study, we confirmed the antitumor activity of axitinib against canine MGT cell lines. Accordingly, we suggest that axitinib can be applied as a new treatment for patients with canine MGTs. [ABSTRACT FROM AUTHOR]

Background/Aim: Nuclear matrix protein-22 (NMP- 22) is widely used in human medicine as a prognostic and diagnostic tool for urothelial carcinoma (UC). In addition, the use of urinary exosomes as a liquid biopsy tool is emerging for the diagnosis of certain types of cancer in human medicine. This study aimed to investigate the change in urinary exosomal NMP-22 for the diagnosis of UC in dogs. Patients and Methods: Among canine patients who visited the veterinary hospital, urine was collected from those whose owners provided consent. A total of 23 dogs (UC group, n=6; control group, n=17) were included in the analysis. After exosomes were isolated from the urine, NMP-22 was measured using enzyme-linked immunosorbent assay. Results: In the UC group, the expression of NMP-22 in urinary exosomes was significantly higher than that in non-UC groups (p<0.0001). Conclusion: NMP-22 is significantly increased in exosomes in the urine of dogs diagnosed with UC, suggesting that urinary exosome NMP-22 can be considered as one of the liquid biopsy tools for diagnosing UC in dogs. [ABSTRACT FROM AUTHOR]

We performed this study to characterize the difference between the inflammatory and non-inflammatory status in diseased dogs by measuring salivary C-reactive protein (CRP) levels. In addition, we assessed whether a correlation exists between CRP levels in saliva and those in serum. CRP levels were measured in 32 client-owned dogs, which were then divided into inflammation and non-inflammation groups based on the serum CRP level. The salivary CRP level was higher in the inflammation group than in the non-inflammation group (p < 0.05). Furthermore, there was a positive correlation between the salivary and serum CRP levels (R = 0.866, p < 0.001). These data suggest that canine salivary CRP measurements can effectively and non-invasively detect an inflammatory state in dogs.

A 2-year-old, spayed female, Bichon Frise dog was presented with reluctance to exercise, back pain, and frequent sitting down. Multiple osteolysis, periosteal proliferation, and sclerosis of the vertebral endplates of T11–13 were observed in the radiography, computed tomography, and magnetic resonance imaging. The bacterial culture of the urine specimen, the polymerase chain reaction (PCR) of the blood, and the antibody tests were positive for Brucella canis. Accordingly, discospondylitis caused by B. canis was diagnosed and doxycycline was administered. The clinical signs resolved and the culture and PCR results were negative afterwards. Doxycycline was discontinued after 6 months. The clinical signs recurred 2 weeks later, and the combination treatment of doxycycline and enrofloxacin was initiated. Though no clinical signs were observed after 9 months and the bacterial cultures and PCR were negative, the antibody titre remained at 1: 200 or more. The dog will continue taking antibiotics until the antibody titre drops. To the best of our knowledge, this is the first case report of a clinical infection of B. canis associated with canine discospondylitis in the Republic of Korea. Although the clinical signs of brucellosis might improve with antibiotic treatment, the disease cannot be cured due to Brucella’s various strategies to evade host immune systems. Specifically, it can proliferate and replicate within the host cells, resulting in an environment that makes treatment less effective. Furthermore, owing to its zoonotic potential, owners and veterinarians should consider lifelong management or euthanasia. [ABSTRACT FROM AUTHOR]