Case report: Allogeneic feline umbilical cord-derived mesenchymal stem cell transplantation for feline oral squamous cell carcinoma.

A 5-year-old neutered female Korean domestic shorthair cat diagnosed with oral squamous cell carcinoma (SCC) presented to the hospital with severe oral purulent discharge, anorexia, and lethargy. Owing to extensive lesions, surgical excision and radiation therapy were not feasible. Instead, prior to metronomic therapy with toceranib, the patient received an intravenous injection of feline umbilical cord-derived mesenchymal stem cells (fUC-MSCs) (1 × 106 cells/10 mL of saline) to reduce inflammation. No acute side effects (such as fever, increased respiratory rate, diarrhea, and vomiting) were observed following stem cell therapy. For 6 days, purulent discharge, bleeding, swelling, a bad odor, and crust exfoliation in the tumor area on the face were dramatically reduced. However, the patient exhibited difficulty in voluntarily receiving foods, and weight loss persisted. Starting from the 7th day, purulent discharge, bleeding, and odor at the SCC area worsened again. Toceranib, low-dose NSAIDs (meloxicam, every other day), antibiotics (cefazoline), and gabapentin were administered; however, they were not effective in reducing the pus, bleeding, foul odor, and crust exfoliation at the SCC area. Symptoms of pain, weakness, and weight loss progressed, leading to the choice of euthanasia with the owner's consent approximately 1 month later. This case report reveals that allogeneic fUC-MSCs have a slight short-term effect on purulent discharge, bleeding, odor, and crust exfoliation and may be additional therapy for feline oral SCC.

Isolation and characterization of feline endometrial mesenchymal stem cells.

BACKGROUND: Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy. OBJECTIVES: This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype. METHODS: Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry. RESULTS: fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 -, CD34 -, CD45 -, CD9+, and CD44+. CONCLUSIONS: In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

Case report: Lymphocytic-plasmacytic and eosinophilic enterocolitis presented with marked eosinophilia and basophilia in a cat.

Inflammatory bowel disease is a common condition in cats, characterized by recurring gastrointestinal signs with histologic evidence of intestinal inflammation. A 9-month-old neutered male Sphynx cat was presented with a 5-week history of vomiting and hematochezia. Conservative patient management with a therapeutic gastrointestinal formula, antibiotics, and antiemetics resulted in a positive response to treatment, with relapse of signs when the medications were discontinued. A new finding of marked eosinophilia and basophilia was identified 3 months after the initial presentation. Colonoscopy revealed cecal erosions and a surgical biopsy with histopathology confirmed a diagnosis of lymphocytic-plasmacytic and eosinophilic enterocolitis. For this diagnosis, the patient was treated with prednisolone, tylosin, and metronidazole. Antibiotics were gradually tapered as the cat showed clinical improvement. The patient showed resolution of the gastrointestinal signs, and the numbers of eosinophils and basophils returned within the reference range 8 weeks after the treatment began. Basophilia and eosinophilia has been reported in conjunction with feline T-cell lymphoma. However, marked basophilia accompanying eosinophilia is extremely rare in cats with inflammatory bowel disease. We herein provide clinical details, including ultrasonography, endoscopy, histopathology, and disease course of feline lymphocytic-plasmacytic and eosinophilic enterocolitis with marked basophilia and eosinophilia. This case highlights the importance of considering enteritis as potential diagnoses when eosinophilia and basophilia are concurrently observed in cats.

Correlation between the Leaflet-Annulus Index and Echocardiographic Indices in Maltese Dogs with Myxomatous Mitral Valve Disease.

Myxomatous mitral valve disease (MMVD) is the most common chronic heart valve disease, leading to left-sided cardiomegaly in dogs. The leaflet-annulus index (LAI) was originally used in humans as a predictor of mitral regurgitation (MR) after mitral valve repair surgery. This index represents the quantity and severity of MR since it is affected by annular dilation. Recently, LAI was adapted to veterinary medicine, and its usefulness as an indicator of annular dilation on 2D transthoracic echocardiography in MMVD dogs was suggested. For this study, 135 Maltese dogs were selected and divided into groups of control, B1, and B2, according to the American College of Veterinary Internal Medicine consensus statement. The following data were collected: radiographic indices including the vertebral heart score and vertebral left atrial size, echocardiographic indices including the left-atrium-to-aortic-root ratio (LA:Ao), left ventricular internal diameter at diastole, normalized for body weight, and anteroposterior length and LAI measured on right parasternal long-axis view. The results showed a significant difference in LAI between each group, becoming smaller as the disease progressed. Also, there was a significant correlation between LAI and each index, showing the strongest correlation with LA:Ao. LAI could be helpful as a new indicator used for the determination of severity and prognosis in Maltese dogs with MMVD.

Case report: Emphysematous cystitis due to Escherichia coli infection with the extension of gas into multiple locations in two non-diabetic dogs: a computed tomographic diagnosis and successful management.

Emphysematous cystitis is an extremely rare, complicated urinary tract infection with the presence of gas in the bladder wall and lumen caused by gas-producing bacterial infections. A 7-year-old spayed female pomeranian dog was presented with a 3-day history of hematuria and pollakiuria (case 1), and a 9-year-old spayed female jindo dog was presented with a 4-day history of intermittent hematuria (case 2). Imaging modalities, including radiography, ultrasonography, and computed tomography, and bacterial culture tests were used for the diagnosis. Emphysematous cystitis due to Escherichia coli infection with the extension of gas into multiple locations was identified in both cases. Based on the results of antibiotic susceptibility testing, systemic antibiotics were initiated. Both animals had an excellent response to antibiotic treatment, and the clinical signs of the gas collection were completely resolved within ~1 month after treatment initiation. This response was sustained without recurrence in the follow-up period. This case report describes clinical details of extremely rare canine cases of emphysematous cystitis with the extension of gas into multiple locations and evaluates the clinical efficacy of antibiotic therapy.

Mott Cell Differentiation in Canine Multicentric B Cell Lymphoma with Cross-Lineage Rearrangement and Lineage Infidelity in a Dog.

Lymphoma is a severe condition characterized by the proliferation of neoplastic lymphoid cells. A 4-year-old female mongrel dog presented with solitary lymph node enlargement. Significant right prescapular lymphadenopathy and abdominal enlargement were observed during physical examination. A complete blood count revealed lymphocytosis, and a peripheral blood smear revealed lymphoblastosis and Mott cells. Fine needle aspiration cytology (FNAC) of the right prescapular lymph node revealed a predominant population of lymphoblasts and Mott cells. Based on the FNAC and blood smear results, the patient was diagnosed with leukemic state multicentric B-cell lymphoma with Mott cell differentiation. Subsequent PCR for antigen receptor rearrangement and flow cytometry revealed that the patient exhibited cross-lineage rearrangement (CLRA) and lineage infidelity (LI), respectively. CHOP-based chemotherapy was initiated, however, the patient's disease was progressive. The patient died three months after the initial presentation. Mott cell differentiation in canine B-cell lymphoma (MCL) has rarely been reported in the veterinary literature and seems to show an unusual clinical course. To the best of our knowledge, no reports of MCL with CLRA and LI exist. We report the clinical features, diagnosis, and treatment of MCL with CLRA and LI.

Treatment of acute erythroleukaemia with high-dose cytarabine in a cat with feline leukaemia virus infection.

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.

Secondary lymphoma development after chemotherapy in three dogs.

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.

Evaluation and comparison of serum procalcitonin and heparin-binding protein levels as biomarkers of bacterial infection in cats.

OBJECTIVES: As bacterial infection can lead to sepsis and high mortality, early and easy diagnosis of sepsis can improve survival. In cats, the diagnosis of systemic bacterial infection is quite challenging, and, usually, non-specific markers for inflammation are employed. In humans, procalcitonin, heparin-binding protein and absolute neutrophil count are biomarkers that are studied in bacterial infections and sepsis owing to their high sensitivity and specificity. METHODS: A total of 56 cats were categorised into 16 healthy cats and 40 bacterially infected cats, diagnosed by various examinations. In all cats, serum procalcitonin and heparin-binding protein levels were measured using ELISA and an absolute neutrophil count was performed. RESULTS: The median values of procalcitonin levels and absolute neutrophil count were significantly higher in the infection group than in the normal group, but heparin-binding protein levels were not. A procalcitonin level >366 pg/ml was a better biomarker of bacterial infection than heparin-binding protein and absolute neutrophil count (sensitivity: 67.5%; specificity: 93.8%). Procalcitonin was not correlated with heparin-binding protein (r = 0.213, P = 0.115) and absolute neutrophil count (r = 0.393, P = 0.003). CONCLUSIONS AND RELEVANCE: High procalcitonin levels in cats were associated with bacterial infection. Hence, procalcitonin could be a valuable marker for diagnosing bacterial infections in cats.

Effect of 7.5% hypertonic saline solution on whole blood coagulation in healthy dogs using thromboelastography.

OBJECTIVE: To evaluate the effects of 7.5% hypertonic saline solution (HSS) on whole blood coagulation in healthy dogs and to compare electrolyte and osmolality measurements between in vivo and in vitro dilution with HSS. DESIGN: Experimental study. SETTING: University teaching hospital. ANIMALS: Twelve adult purpose-bred Beagles. INTERVENTIONS: All 12 dogs received 5 mL/kg 7.5% HSS at 1 mL/kg/min. After a 14-day washout period, 5 of these dogs were randomly selected and received the same volume of 0.9% NaCl. Blood samples were collected before infusion, immediately after infusion, and at 30, 60, and 90 minutes after infusion for the measurement of coagulation using thromboelastography. For comparison of electrolyte concentrations and osmolality between in vitro dilution and in vivo dilution of HSS, 6-mL blood samples were diluted with 7.5% HSS (1:18 ratio) at baseline. MEASUREMENTS AND MAIN RESULTS: None of the thromboelastography variables differed significantly between the 7.5% HSS group and the 0.9% NaCl group. The sodium and chloride levels, and the osmolality, were significantly increased at all postinfusion time points compared to baseline, while those levels were significantly higher with in vitro dilution than all postinfusion time points. However, almost all the values gradually decreased and became similar to baseline values in case of in vivo dilution. CONCLUSIONS: The clinically relevant dose of 7.5% HSS (5 mL/kg) did not affect whole blood coagulation significantly in healthy Beagles. Further studies are necessary to assess the effect of HSS on blood coagulation in canine patients with shock.

Pharmacokinetics of intravenous administered two different high doses of ascorbic acid in healthy beagle dogs.

OBJECTIVE: We performed a randomized two-way crossover study to evaluate the pharmacokinetic profiles of two high-dose ascorbic acid (AA) after IV infusion in healthy beagle dogs. MATERIALS AND METHODS: The dogs were administered IV AA at two doses of 1.5 and 3 gm/kg for 4 h, and the AA concentration in plasma and urine pH was measured before and after administration. RESULTS: The plasma concentrations of AA in both groups peaked 3 h after administration. Among the two groups, the urine pH was not significantly different (p = 0.1299-0.7944). High-dose IV AA did not induce serious adverse events in dogs. CONCLUSION: The results of this study suggest that the high dose of AA which reaches the therapeutic dose for cancer and supports the safety of high-dose IV AA in dogs.

Influence of needle gauge and venepuncture difficulty on thromboelastography in healthy cats.

OBJECTIVES: The aim of this study was to evaluate the effect of two differently sized butterfly catheter needles and the effect of venepuncture difficulty on thromboelastography (TEG) results in healthy cats. METHODS: Twenty-four healthy cats were included. Blood samples were collected from the jugular vein by syringe aspiration via direct venepuncture with 21 G and 22 G butterfly needles. The venepuncture difficulty score was classified into four categories. The first 1.5 ml blood drawn from each subject was discarded before collecting a sample for TEG analysis. TEG analyses were performed on citrated whole blood samples from 17 clinically healthy cats, using assays with kaolin as activators. Among the TEG parameters, reaction time (R), clot formation time (κ), alpha angle (α), maximum amplitude (MA) and global clot strength (G) were recorded from each tracing. RESULTS: Seven cats were excluded from the study; results were obtained for the remaining 17 cats. There were no statistically significant differences between the use of two different needles for R (P = 0.72), κ (P = 0.74), α (P = 0.99), MA (P = 0.08) and G (P = 0.09). Samples with difficulty scores ⩾1 were not significantly different from samples with difficulty scores of 0 for R (P = 0.24), κ (P = 0.65), α (P = 0.65), MA (P = 0.72) and G (P = 0.77). CONCLUSIONS AND RELEVANCE: The results of TEG in clinically healthy cats do not differ significantly when using two different gauge needles. There was no significant difference in the TEG results according to venepuncture difficulty scoring.

Anticancer effects of high-dose ascorbate on canine melanoma cell lines.

The use of the well-known powerful antioxidant ascorbate has recently become more widespread in human medicine. Intravenous administration of high-dose ascorbate has been demonstrated to exert anticancer effects. It has resulted in effective cell death in vitro and inhibition of tumour growth in vivo. The aim of the current study was to evaluate the effects of high-dose ascorbate on canine melanoma in vitro. Four canine melanoma cell lines, UCDK9M1, UCDK9M3, UCDK9M4 and UCDK9M5 were treated with ascorbate for 2 hours at a range of millimolar concentrations (0-20 mM) to investigate the resulting effects on cell viability. All four canine melanoma cell lines exhibited reduced viability in a dose-dependent manner. Further investigation demonstrated that high-dose ascorbate induced apoptosis via the activation of Bax. These findings suggest that high-dose ascorbate has an anticancer effect on canine melanoma cell lines in vitro. With regard to clinical application, further in vivo investigation should be conducted.

Application of vincristine-loaded platelet therapy in three dogs with refractory immune-mediated thrombocytopenia.

Three dogs presented with refractory immune-mediated thrombocytopenia (IMT). All patients failed to respond to prednisone, which is considered a mainstay of immunosuppressive therapy. Vincristine-loaded platelets (VLPs), which act selectively on mononuclear phagocytes,were introduced. After the VLPs were transfused, two dogs responded quickly with improved clinical signs while the third dog with recurrent IMT was euthanized due to its deteriorating condition. This case report describes the efficacy of VLP therapy in refractory IMT patients.

The effect of oculo-acupuncture on recovery from ethylene glycol-induced acute renal injury in dogs.

The potential recovery effect by oculo-acupuncture (OA) on ethylene glycol-induced acute renal injury in dogs was investigated. Acute renal damage was induced by ingestion of ethylene glycol in six mongrel dogs. The dogs were assigned to control (three dogs) and experimental (three dogs) groups. The control group did not receive any treatment, while the experimental group was treated with oculo-acupuncture at kidney/urinary bladder region plus zhong jiao region of the eyes after the induction of renal damage. Serum blood urea nitrogen (BUN), creatinine, sodium (Na), chloride (Cl), and potassium (K) were measured in both control and experimental groups. The blood RBC and Hb were also examined. The serum BUN and creatinine activities in the experimental group were lower than those in the control group, the serum Na and Cl had the irregular change in both groups, and the blood Hb in the control and experimental group showed decreasing tendency. Significant differences were observed on the 3rd and 7th day in BUN, 7th day in creatinine, 2nd day in Na and Cl, and 7th day in Hb when compared to the control group. Whereas, serum K concentration and RBC in the experimental group did not change significantly. The recovery findings of the renal injury were also observed in the experimental group histopathologically. In conclusion, OA therapy (kidney/urinary bladder region plus zhong jiao region) was effective for recovery of the renal injury induced by ethylene glycol in dogs.

Electroacupuncture analgesia for surgery in cattle.

To establish the proper analgesic method by electroacupuncture (EA) for bovine surgery, the analgesic effect of dorsal and lumbar acupoints, in addition to the combination with dorsal and lumbar acupoints, were investigated in the present study. Four Korean native cattle (two males and two females) and 24 Holstein-Friesian cattle (all females) were used. The experimental animals were divided into four groups according to used acupoints: dorsal acupoint group (Tian Ping [GV-20] and Bai Hui [GV-5]: 7 heads), lumbar acupoint group (Yap Pang 1 [BL-21], Yao Pang 2 [BL-23], Yao Pang 3 [BL-24] and Yao Pang 4 [BL-25]; 5 heads), dorsal-lumbar acupoint group (Yao Pang 1 [BL-21], Yao Pang 2 [BL-23], Yao Pang 3 [BL-24] and Bai Hui [GV-5]; 8 heads) and control group (non-acupoints, the last intercostals space and the femoral area; 3 heads). The acupoints were stimulated with currents of 2-6 V (30 Hz) in dorsal acupoint group, 0.5-2.0 V (30 Hz) in lumbar acupoint group and 0.3-2.5 V (30 Hz) in dorsal-lumbar acupoint group. Recumbency time was 10 seconds to 1 minute (except one case) and induction time of analgesia was approximately 1 to 6 minutes in dorsal acupoint group. Analgesic effect was systemic, including the extremities in dorsal acupoint group. During the EA, the consciousness was evident and blepharo-reaction was still present under EA in dorsal acupoint group. During the surgery, grades of analgesic effect were 6 excellent (6/7, 87.5%) and 1 good (1/7, 14.3%). In addition, induction time for analgesia was about 10 minutes in both lumbar and dorsal-lumbar acupoint groups. Analgesic areas were found in abdominal areas from the last intercostal spaces to the femoral areas, except lower abdomen in lumbar and lumbar-dorsal acupoint groups. The consciousness was evident and standing position was maintained during EA stimulation in contrast to that of dorsal excellent (1/5, 20.0%), 3 good (3/5, 60.0%) and 1 poor (1/5, 20.0%) in the lumbar acupoint group. Additionally, grades of analgesic effect were 4 excellent (4/8, 50.0%), 3 good (3/8, 37.5%) and 1 poor (1/8, 12.5%). On the other hand, pain was present and analgesia was not accomplished under EA stimulation in control group. In conclusion, analgesia by EA was effective with decreasing order of dorsal acupoint > dorsal-lumbar acupoint > lumbar acupoint among groups. It was considered that dorsal acupoint group might be useful for operation with recumbent position, and lumbar and dorsal-lumbar acupoint groups might be proper for operation with standing position.