Long-term treatment of allogeneic adipose-derived stem cells in a dog with rheumatoid arthritis.

BACKGROUND: Although there are growing demands for stem cell-based therapy for companion animals in various diseases, a few clinical trials have been reported. Moreover, most of them are the results from only one or a few times of stem cell injection. OBJECTIVES: The aim of this study is to describe a long-term treatment with allogeneic adipose-derived stem cells (ASCs) in a dog with rheumatoid arthritis (RA), which is a rare canine disease. METHODS: The dog with RA received intravascular injection of allogeneic ASCs derived from two healthy donors once a month for 11 months. To assess therapeutic effects of ASCs, orthopedic examination and clinical evaluation was performed. Cytokines of tumor necrosis factor-α and interleukin-6 in the plasma were measured using ELISA analysis. RESULTS: Despite this repeated and long-term administration of allogeneic ASCs, there were no side effects such as immunorejection responses or cell toxicity. The orthopedic examination score for the dog decreased after ASCs treatment, and the clinical condition of the dog and owner's satisfaction were very good. CONCLUSIONS: Although ASCs has been suggested as one of the options for RA treatment because of its anti-inflammatory and immunosuppressive functions, it has never been used to treat RA in dogs. The present report describes a case of canine RA treated with allogeneic ASCs for long-term in which the dog showed clinical improvement without adverse effects.

Effect of nabumetone on humoral immune responses in mice / Efeito da nabumetona nas respostas imunes humorais em camundongos

Nabumetone is used to reduce the pain and inflammation in rheumatoid arthritis. In the current study, immunomodulatory effect of Nabumetone is investigated in mice. The control group was administered normal saline orally as placebo. Nabumetone was administered orally via gavage in two treatment groups at 14mg/kg.b.w. doses and 28mg/kgb.w., respectively. Haemagglutination (HA) assay, Jerne hemolytic plaque and mice lethality assays were applied. In HA assay, the titer was significantly decreased in Nabumetone treatment groups (P< 0.001). In Jerne hemolytic plaque formation assay, there was a significant reduction (P< 0.001) in number of plaques in Nabumetone treated groups when compared with control. In mice lethality assay, there was a significant difference in mortality ratio of mice in control and Nabumetone treated groups (P< 0.001). Therefore, it is concluded that Nabumetone suppresses the humoral immune response in mice.(AU)

A nabumetona é usada na redução da dor e inflamação da artrite reumática. No presente estudo, o efeito imunomodulador é investigado em camundongos. O grupo de controle recebeu solução salina via oral como placebo. Nabumetona foi administrada oralmente via gavagem em dois grupos de tratamentos com doses de 14mg/kg.b.w. e 28mg/kgb.w., respectivamente. Foram realizados ensaios de hemaglutinação (HA), placa hemolítica de Jerne e letalidade dos camundongos. No ensaio HA, o grau foi significativamente menor nos grupos de tratamento com nabumetoma (P< 0.001). No ensaio de formação de placa hemolítica de Jerne houve redução significativa (P< 0.001) no número de placas em grupos tratados com nabumetoma comparado ao controle. No ensaio de letalidade dos camundongos houve diferença significativa no grau de mortalidade de camundongos no grupo de controle e grupos tratados com nabumetoma (P< 0.001). Portanto, conclui-se que a Nabumetoma suprime a resposta imune humoral em camundongos.(AU)

Comparison of the efficacy of prednisone and cyclosporine for treatment of dogs with primary immune-mediated polyarthritis.

OBJECTIVE: To compare efficacy between cyclosporine and prednisone for treatment of primary immune-mediated polyarthritis (IMPA) in dogs. DESIGN: Randomized controlled clinical trial. ANIMALS: 20 client-owned dogs with primary IMPA. PROCEDURES: Dogs were randomly assigned to receive prednisone (starting at 1 mg/kg [0.45 mg/lb], PO, q 12 h; n = 10) or cyclosporine (5 mg/kg [2.3 mg/lb], PO, q 12 h; 10) for 90 days. Cyclosporine-treated dogs also received carprofen, tramadol, or both for the first 7 days for analgesia. Data collection, physical examination, and cytologic analysis of synovial fluid samples were performed on days 0, 14, 45, and 90. Trough whole blood cyclosporine concentrations were determined on days 7 to 17 for cyclosporine-treated dogs. Treatment failure was defined as lack of clinical improvement by day 14, lack of cytologic improvement by day 45, or need to change treatment because of adverse effects. RESULTS: Treatment was successful for 7 prednisone-treated dogs and 7 cyclosporine-treated dogs. Absence of synovial fluid cytologic abnormalities on day 45 was identified for 5 prednisone-treated dogs and 8 cyclosporine-treated dogs. Prednisone-treated dogs were more likely to develop polyuria, polydipsia, and polyphagia than were cyclosporine-treated dogs. Opportunistic infections (ie, demodicosis or Erysipelothrix bacteremia) were identified in 2 cyclosporine-treated dogs and 0 prednisone-treated dogs, and diarrhea developed in 1 cyclosporine-treated dog, requiring treatment discontinuation. CONCLUSIONS AND CLINICAL RELEVANCE: Although the number of dogs evaluated was small, limiting generalizability, results of this study suggested that cyclosporine offers promise as a suitable alternative to prednisone for treatment of IMPA in dogs.

Disease modifying treatment for feline rheumatoid arthritis.

Feline erosive polyarthritis includes the more common periosteal proliferative polyarthritis (PPP) and the rarely seen rheumatoid arthritis (RA) (11). During the past three years, 12 patients with definite feline rheumatoid arthritis, which did not respond well to conventional therapy, were treated with 7.5 mg of Methotrexate and 70 mg Leflunomide, given weekly by the oral route. The average age of the cats was 5.9 years (range 2.5 to 10 years). Siamese cats were over represented. Seven of the 12 (58%) cats showed a marked improvement, usually within four weeks. Once maximum improvement was obtained the dosage was decreased. Serious toxicity was not noted and carcinogenetic effect was not seen during the course of this study.

Synovial MMP-3 and TIMP-1 levels and their correlation with cytokine expression in canine rheumatoid arthritis.

The purpose of this study was to investigate whether synovial fluid levels of matrix metalloprotease-3 (MMP-3) and tissue inhibitor of metalloprotease-1 (TIMP-1) are specifically elevated in canine rheumatoid arthritis (CRA) compared to osteoarthritic joint disorders and if these markers are correlated with a specific pattern of cytokine mRNA expression. Synovial fluid samples of 17 dogs with CRA were analysed for MMP-3 and TIMP-1 by two canine sandwich ELISA (enzyme-linked immunosorbent assay) systems. The synovial mRNA content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-12 (IL-12), transforming growth factor-beta (TGF-beta), and interferon-gamma (IFN-gamma) was determined by RT-PCR (reverse transcription-polymerase chain reaction). Dogs with osteoarthritis (n = 50) caused by anterior cruciate ligament rupture (ACLR) were used as controls. A significant rise of MMP-3 was found in the synovial fluid of joints with CRA that could not be balanced by sufficient amounts of TIMP-1. The 30-fold surplus of MMP-3 over TIMP-1 was strongly correlated with the synovial mRNA content of IL-1, IL-12 and TGF-beta. Our results point to the potential use of the synovial levels of MMP-3 as a marker for RA in dogs.

Canine rheumatoid arthritis and inflammatory cytokines.

Bioassays were developed to detect canine pro-inflammatory cytokines. These enabled characterisation of these cytokines and their isoforms and provided means for their assay in the joints of dogs with different naturally occurring arthropathies. Canine IL-1 was detected by its induction of proliferation of D10(N4)M cell line, whilst IL-6 had a proliferative effect on B9 cell line. TNFalpha had a cytotoxic effect on WEHI 164 (13) cells. Partial purification of the cytokines was achieved by FPLC ion-exchange chromatography and two isoforms of IL-1 were shown, possibly corresponding to IL-1alpha and IL-1beta. TNFalpha only appeared as one isoform whereas IL-6 showed at least five isoforms, possibly corresponding to the other molecules in the IL-6 family, such as IL-11 and oncostatin M. Analysis of synovial fluids from dogs with osteoarthritis (OA) and rheumatoid arthritis (RA) showed that IL-1 and TNFalpha bioactivity was not readily detectable at increased levels in diseased joints but that IL-6 was significantly increased in both diseases. It is now important to determine the role of IL-6 in OA and RA in the dog, particularly in the induction of proteolytic enzymes which lead to cartilage loss.

Hyaluronan in canine arthropathies.

Soluble hyaluronan (HA), which has been considered as a marker for joint disease in man, was measured in serum and synovial fluid (SF) from dogs with osteoarthritis (OA), rheumatoid arthritis (RA), and rupture of the cranial cruciate ligament (CCL) and from normal dogs (control). Dogs with OA and RA had significantly increased serum HA (P < 0.001) and decreased synovial fluid HA (P < 0.001), as did dogs with CCL rupture (serum, P < 0.05; synovial fluid, P < 0.005). In OA, HA was lower in the SF from the affected joint than in that from the clinically normal (inactive) contralateral joint; no such difference was seen in dogs with CCL rupture. Dogs with liver disease (portocaval shunts, viral infectious hepatitis, metastatic neoplasm and disease secondary to diabetes mellitus) had increased serum HA concentrations (P < 0.001). There was a significant overlap of HA values in the diseased and normal dogs. Therefore, it is unlikely that the measurement of this cartilage breakdown product would be of value for diagnosis or prognosis in canine arthropathies.

Lymphocyte populations in the synovial membranes of dogs with rheumatoid arthritis.

Lymphocyte populations in the synovial membranes of dogs with canine rheumatoid arthritis (CRA) were investigated by immunohistochemical staining techniques. T-lymphocytes were the predominant cell type distributed throughout the supporting layer of the synovial membranes. B-lymphocytes expressing IgG were seen far more commonly than those expressing either IgA or IgM. Synovial membrane biopsies from normal and osteoarthritic joints did not have the marked cellular infiltrates seen in joints with CRA. The synovial immunohistopathological features in dogs with CRA are similar to those seen in human rheumatoid arthritis.

C-reactive protein in dogs.

Serum concentrations of C-reactive protein (CRP) in dogs with various diseases or undergoing various procedures were measured by specific immunoassay. In 20 healthy dogs from various sources, values were all less than 5 mg/L, but in 22 healthy dogs from a single source, values ranged from less than 5 mg/L in 14 dogs and from 8 to 67 mg/L in 8 dogs. Increased concentrations of serum CRP were attained 24 hours after injection of casein (n = 9; median 188 mg/L), ovariohysterectomy (n = 11; median, 144 mg/L), or elective, nonacute orthopedic surgery (n = 10; median, 83 mg/L). After inoculation of Leptospira interrogans serovar canicola (n = 5), the behavior of serum CRP as an acute-phase reactant provided a sensitive and precise objective reflection of in vivo response. The CRP concentration in random single-serum samples from 73 dogs with other inflammatory and noninflammatory disorders ranged from normal (less than 5 mg/L) to 246 mg/L and generally correlated with the extent and activity of disease.

Synovial pathologic changes in spontaneous canine rheumatoid-like arthritis.

The synovial fluid and membrane were studied in 10 dogs meeting the American Rheumatism Association criteria for classic human rheumatoid arthritis (RA). Light microscopic pathologic features were consistent with those found in the human disease. Neutrophilic infiltration of synovium was somewhat more prominent than in chronic human RA, and activated lymphocytes in fluid or membrane were less frequent. The proliferative and plasma cell reaction seemed identical. Electron microscopy (EM) suggested microvascular injury with findings which included electron dense deposits in the vessel walls of 2 dogs. Seven dogs had meshworks of 20--25 nm tubules in tubuloreticular structures (TRS) similar to those seen in human systemic lupus erythematosus and only occasionally in human RA. There were also crystalline arrays of tubules, a configuration previously reported in tumors and virus infections and possibly suggestive of a cellular reaction to virus infection. To date no initiating agent has been identified, but this spontaneous canine disease which is very similar to human RA can provide a valuable model in which to examine pathogenesis of chronic arthritis.

Amyloidosis in rhesus monkeys with rheumatoid arthritis and enterocolitis.

Generalized amyloidosis was diagnosed by light and electron microscopy in 4 of 5 monkeys (Macaca mulatta) that had a history of chronic arthritis or chronic intermittent diarrhea, or both. Clinical signs and lesions of arthritis in the monkeys were compatible with diagnostic criteria for rheumatoid arthritis. Shigella sp was cultured from 1 monkey, and 2 other monkeys had colonic lesions characteristic of shigellosis. At necropsy, gross changes attributed to amyloid were seen only in the liver. Amyloid deposits in the liver, spleen, mesenteric lymph nodes, kidney, heart, adrenal glands, and other organs were determined by staining reactions and fine structural characteristics.

Application of the radiological sciences to advance clinical and fundamental knowledge of the animal kingdom. New Horizons for Radiologists Lecture.

Many radiological techniques are useful in fundamental research. Selected clinical problems in animals as well as certain basic research projects are utilized to emphasize the potential of radiological imaging. The author describes the use of several radiological methods in studies of how birds breathe.

[Clinical data on the use of estriol-testosterone combinations in small animal practice]. / Klinische Daten bei der Verwendung von Ostriol-Testosteron-Kombinationen in der Lleintierpraxis

A combination of oestriol and testosterone oenanthate was tried in 14 different indications on 110 dogs and 5 cats of both sexes. Two formulations were used, containing in 1 ml either 0.15 mg oestriol and 2.25 mg testosterone or 1.15 mg of the former and 2.25 mg of the latter. Dosage was 0.2 ml per kg body weight, by i/m injection. The criterion for success was the degree of functional restitution, without taking into account pathological changes, which were generally unaffected. Good or very good results were obtained in hip dysplasia, arthitis and patellar luxation. None of the 50 bitches treated showed signs of oestrus or irregularities in the cycle. The formulation containing the lower drug concentration seemed to give the best therapeutic results.