Osteoarthritis (OA) can be debilitating and career-ending for horses. Current treatments offer temporary and symptomatic relief, but potentially deleterious side effects. Bone marrow mononuclear cells (BMNC) are a rich source of macrophage progenitors that are anti-inflammatory and promote inflammation resolution. The objective of this study was to evaluate the ability of intra-articular BMNC therapy to improve clinical signs of naturally occurring equine OA. Horses presenting with clinical and radiographic evidence of moderate OA in a single joint were randomly assigned to 1 of 3 treatments: saline (negative control), triamcinolone (positive control), or BMNC (treatment group). Horses were subjectively and objectively evaluated for lameness and synovial fluid collected (cytology and cytokine/growth factor quantification) at 0, 7, and 21 days post-injection. Data were analyzed using General Estimating Equations with significance set at P<0.05. There were no adverse effects noted in any treatment group. No significant differences in synovial fluid cytology parameters, objective/subjective lameness scores, nor joint circumference were found between treatment groups at any time point. Within treatment groups, joint circumference did not change over time for saline- and triamcinolone-treated horses. However, joint circumference and objective lameness decreased significantly within BMNC-treated horses between Days 0 and 21 and Days 7 and 21. Lameness improved in saline-treated horses from 0 to 21 days, but did not improve in triamcinolone-treated horses. The decreased lameness and lack of adverse effects in the BMNC-treated horses in our study support a larger clinical trial using BMNC. / Master of Science / Osteoarthritis (OA) is a common source of joint pain in people and horses. Current treatments provide only partial and/or temporary relief. As a result, there is an urgent need for more effective and long-lasting treatment options. Arthritis is characterized by uncontrolled joint inflammation and progressive cartilage and bone destruction. Macrophages are cells within normal joints that function to resolve mild inflammation, maintaining joint health. However, when physiologic functions are overwhelmed, macrophages perpetuate inflammation through the recruitment of additional cell types to cope with the increased demands for repair. If this process is appropriately accomplished, macrophages resolve the inflammation, thereby enabling recovery and repair within the joint. Bone marrow aspirate is an excellent source of bone marrow-derived macrophage precursors (bone marrow mononuclear cells or BMNC) that have been shown to reduce joint inflammation and lameness in people and horses. The objective of our clinical trial was to evaluate the ability of intra-articular BMNC to improve clinical signs of naturally occurring OA in horses. BMNC treatment was compared to a placebo injection of saline and a standard-of-care in horses, corticosteroids. There were no adverse effects of BMNC treatment and BMNC-treated horses had significantly reduced joint circumference and lameness after 21 days. Synovial fluid cytology parameters did not differ significantly between treatment groups at any time point. In summary, BMNC are exciting because a horse can be treated with its own cells without the need for specialized equipment, and have the potential to naturally benefit thousands of people and horses suffering from arthritis.

Latent feline leukemia virus (FeLV) infections, in which proviral DNA is integrated into host DNA, but not actively transcribed, are suspected to be associated with many diseases. Bone marrow is the suspected site of the majority of latent infections. The purpose of this study was to determine if polymerase chain reaction (PCR) could detect FeLV proviral DNA in bone marrow and provide a method of detecting latent infections. Blood and bone marrow samples from fifty cats and bone marrow from one fetus were collected; sixteen had FeLV-associated diseases. Serum ELISA, blood and bone marrow immunofluorescent antibody test (IFA), and blood and bone marrow PCR were performed on each cat, and IFA and PCR on bone marrow of the fetus. Forty-one cats were FeLV negative. Five cats and one fetus were persistently infected with FeLV. Four cats were discordant; two ELISA positive with other tests negative, one bone marrow IFA negative with other tests positive, and one bone marrow IFA positive with other tests negative. No cats were positive on bone marrow PCR only. These results indicate that PCR can detect FeLV in bone marrow, but no cats in this study harbored FeLV only in the bone marrow. Not all cats with FeLV-associated diseases are persistently or latently infected with FeLV. / Master of Science

Thesis (doctoral)--Katholieke Universiteit te Nijmegen.

Source: Dissertation Abstracts International, Volume: 70-07, Section: B, page: 4063. Adviser: David R. Lairson. Includes bibliographical references.

Orientador: Lígia Niero-Melo / Banca: Lucilene Silva Ruiz e Rezende / Banca: Eduardo Moreira de Queiroga / Resumo: A infiltração de MO por tumores não-hematológicos tem sido descrita desde 1935 e, mais recentemente, desperta interesse da comunidade científica, pois inúmeros trabalhos têm relacionado a presença de células tumorais na MO com o processo de carcinogênese e a progressão tumoral. O presente estudo teve por objetivo o levantamento dos casos com diagnóstico de mielopatia infiltrativa por tumores nãohematológicos, provenientes da Faculdade de Medicina de Botucatu - UNESP e do Hospital Amaral Carvalho - Jaú, avaliados, retrospectivamente, no período de 1998 a 2008 (num universo de 15.191 coletas de MO). Incluímos 193 pacientes, dos quais 171 eram adultos e 22 crianças (idade ≤ 21 anos), com paridade entre os sexos (1:1) e mediana de idade de 55 anos. Foram analisados dados clínicos, diagnóstico e estadiamento do tumor primário, sítios de metástases, características laboratoriais (hemograma e marcadores tumorais séricos), características da MO, tratamento e a relação desses fatores com a sobrevida dos pacientes, através do programa de estatística SPSS®. Os resultados encontrados apontam que os tumores que mais frequentemente infiltraram a MO nos adultos foram: tumores de mama, próstata e sítio primário indeterminado, e neuroblastoma nas crianças. A análise da MO revelou positividade de 80% para mielograma e BMO, e presença de fibrose em 32% dos pacientes (40%, quando associados a áreas de necrose no mesmo material). Na avaliação hematológica observamos: anemia, alteração de leucócitos e plaquetopenia. Houve diferença, estatisticamente significante, com maior mediana de sobrevida global (p= 0,000) para pacientes do sexo feminino, portadores de adenocarcinoma de mama, em estádios iniciais e que receberam algum tipo de tratamento (quimio, radio e/ou hormonioterapia). Na análise de sobrevida, a partir do diagnóstico de mielopatia infiltrativa, houve... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Bone marrow (BM) infiltration by nonhematopoietic tumors has been described since 1935, and a increased number of papers about this topic have been published. Some of the studies have related the relationship between the presence of peripheral circulating tumors cells, bone marrow infiltration, carcinogenesis and tumor progression. This study has, as essential objective, the evaluation of infiltrative mielopathy by nonhematopoietic tumors from Faculdade de Medicina de Botucatu - UNESP and Hospital Amaral Carvalho - Jaú. Data were analised retrospectively from 1998 January to 2008 August (15.191 bone marrow samples). We have included 193 patients. One hundred seventy one were adults and 22 were children (considered age ≤ 21 years), with similar number of patients by gender, and median age 55 years. Medical records from the selected patients were analised, with focus on clinical aspects, primary site of tumor, initial stage, metastatic sites, hematologic and bone marrow features, treatment and the relationship between this data and survival. Results have demonstrated that most frequent solid tumors in adults were breast cancer, prostate cancer and undetermined primary site, and neuroblastoma among children. Bone marrow revealed infiltration in 80% of samples by myelogram and BM biopsy, and fibrosis in 32% (40% when associated with necrosis area). We observed anaemia, leukocyte alterations and thrombocytopenia as hematologic features. There were significant differences in global survival, wich was better among female patients, diagnosis of breast cancer, initial clinical stages and who has received any type of treatment (chemotherapy, radiotherapy and/or hormoniotherapy). When we analised survival since diagnosis of infiltrative mielopathy, adult patients with leucoeritroblastic reaction and trombocytopenia, and undetermined primary site tumors, melanoma or gastrointestinal tumors had... (Complete abstract click electronic access below) / Mestre

Deficiency in β-glucuronidase activity (EC 3.2.1.31) leads to the lysosomal storage disease mucopolysaccharidosis type VII not only in humans but also in a recently discovered murine mutant, the gus^mps/gus^mps mouse. Clinical and pathologic abnormalities common to the human and mouse phenotypes include shortened life span, dwarfism, dysmorphic facial features, skeletal deformities, corneal clouding, mental retardation and abnormal lysosomal storage material in the brain and peripheral organs. In the first part of this thesis, neonatal gus^mps/gus^mps mice and their normal littermates were transplanted with syngeneic normal bone marrow. Neurological function was then evaluated with two behavioral tests: the grooming test, a developmentally regulated and genetically based activity, and the Morris water maze test, which assessed spatial learning abilities. The results of these tests indicated that the behavioral deficits in the mutant mice were not restored to normal. Treated normal mice also showed significant functional deterioration, indicating the detrimental consequence of this therapy in the neonatal period. The second part of this thesis focused on a novel approach to somatic gene therapy using microcapsules. A non-autologous fibroblast cell line engineered to secrete high levels of β-glucuronidase was enclosed in perm-selective and immuno- protective microcapsules and implanted into the peritoneal cavity of gus^mps/gus^mps mice. During the 4 weeks of therapy, the biochemical and histological abnormalities of the mutant mice had significantly improved. β-Glucuronidase activity was restored to >50% of normal in the plasma and 11.3%-65.8% in the kidney, liver and spleen. No significant activity was found in the brain. As well, the secondary elevations of other lysosomal enzymes such as β-hexosaminidase and α-galactosidase had decreased in the kidney, liver, and spleen. Urinary glycosaminoglycan content had decreased in the treated mutants indicating that the β-glucuronidase was exerting a therapeutic effect. However, after three and a half weeks of therapy, the treated mutants became severely ill and developed haemorrhagic ascites. Since normal mice treated with similar microcapsules showed no adverse effects, we hypothesized that an immune response had been generated against the foreign protein (β-glucuronidase) by the mutants, leading to the high morbidity. Thus in spite of the biochemical and histological correction observed after bone marrow transplantation and somatic cell gene therapy, the long term efficacy of these treatments needs to be further evaluated. / Thesis / Master of Science (MS)

Human cells from acute myeloid leukemia (AML) patients are frequently transplanted into immune-compromised mouse strains to provide an in vivo environment for studies on the biology of the disease. Since frequencies of leukemia re-initiating cells are low and a unique cell surface phenotype that includes all tumor re-initiating activity remains unknown, the underlying mechanisms leading to limitations in the xenotransplantation assay need to be understood and overcome to obtain robust engraftment of AML-containing samples. We report here that in the NSG xenotransplantation assay, the large majority of mononucleated cells from patients with AML fail to establish a reproducible myeloid engraftment despite high donor chimerism. Instead, donor-derived cells mainly consist of polyclonal disease-unrelated expanded co-transplanted human T lymphocytes that induce xenogeneic graft versus host disease and mask the engraftment of human AML in mice. Engraftment of mainly myeloid cell types can be enforced by the prevention of T cell expansion through the depletion of lymphocytes from the graft prior transplantation.

Thesis (M.S.)--University of Michigan, 1996.

Thesis (M.S.)--University of Michigan, 1996.

Human cells from acute myeloid leukemia (AML) patients are frequently transplanted into immune-compromised mouse strains to provide an in vivo environment for studies on the biology of the disease. Since frequencies of leukemia re-initiating cells are low and a unique cell surface phenotype that includes all tumor re-initiating activity remains unknown, the underlying mechanisms leading to limitations in the xenotransplantation assay need to be understood and overcome to obtain robust engraftment of AML-containing samples. We report here that in the NSG xenotransplantation assay, the large majority of mononucleated cells from patients with AML fail to establish a reproducible myeloid engraftment despite high donor chimerism. Instead, donor-derived cells mainly consist of polyclonal disease-unrelated expanded co-transplanted human T lymphocytes that induce xenogeneic graft versus host disease and mask the engraftment of human AML in mice. Engraftment of mainly myeloid cell types can be enforced by the prevention of T cell expansion through the depletion of lymphocytes from the graft prior transplantation.