Bacterial infection within a joint, septic arthritis, is a serious condition in horses that can lead to long-term joint disease if the infection is not resolved quickly. Equine septic arthritis is diagnosed primarily based on clinical signs and synovial fluid cytology. Septic synovial fluid is characterized by significant elevations in total protein (TP) and total nucleated cell count (TNCC). However, in some cases it can be difficult to distinguish between septic arthritis and non-septic joint inflammation (synovitis) based on clinical signs and synovial fluid cytology alone. A rapid assay to help confirm septic arthritis would be advantageous. A new assay to quantify the major equine acute phase protein, serum amyloid A (SAA) may fulfill this need. Serum amyloid A increases in the body in response to injury, infection, and inflammation and shows promise as a useful tool in confirming a diagnosis of sepsis, as inflammation causes mild increases in SAA and infection causes marked elevations. In our study, serial serum and synovial fluid samples were collected from horses with experimental models of synovitis and septic arthritis, synovial fluid cytology was performed, and serum and synovial fluid SAA were quantified. Synovial fluid TNCC and TP concentrations increased significantly following induction of both models. Serum and synovial fluid SAA concentrations remained normal in synovitis horses and increased significantly in septic arthritis horses. Any elevation in serum or synovial fluid SAA above normal values may be supportive of synovial sepsis since synovial inflammation alone did not result in SAA elevations in our model. / Master of Science

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Orientador: José Jurandir Fagliari / Banca: Rafael Resende Faleiros / Banca: Delphim da Graça Macoris / Banca: José Dantas Ribeiro / Banca: Daniela Gomes da Silva / Resumo: Este estudo teve como objetivo geral comparar os efeitos da laparotomia exploratória e do uso da fenilbutazona em bovinos e bubalinos. Para isso foram avaliados o perfil bioquímico, o proteinograma e o hemograma de 28 animais dos quais 14 eram búfalas e 14 vacas, todas fêmeas não lactantes, distribuídas em quatro grupos - VT: vacas tratadas com fenilbutazona; VC: vacas controle; BT: búfalas tratadas com fenilbutazona e BC: búfalas controle. Os animais dos grupos VT e BT receberam dose única de 10 mg/kg de fenilbutazona, por via intravenosa, uma hora antes da cirurgia. Foram coletadas amostras de sangue antes da laparotomia (0h) e 1(1h), 6(6h), 12(12h), 24(24h), 48(48h), 72(72h), 96(96h) e 120 horas (120h) e 15(15d) e 30(30d) dias após a intervenção. Os valores de creatinina foram mais elevados nas búfalas, e não sofram alteração devido à cirurgia, ou ao tratamento. O tratamento com fenilbutazona aumentou a concentração sérica de HDL em vacas e búfalas. Dentre as proteínas de fase aguda, a concentração de ceruloplasmina se elevou em todos os grupos de 48 horas a 120 horas após a cirurgia. A concentração sérica de haptoglobina se elevou mais nos grupos das búfalas do que nas vacas. A hemopexina foi verificada apenas na espécie bubalina e apenas até o 5º dia. O tratamento com fenilbutazona não alterou a contagem de hemácias nem o volume globular. Todas as vacas e búfalas apresentaram neutrofilia 12 horas após a laparotomia. O teor de fósforo aumentou no grupo controle das búfalas (BC) e no grupo tratado (BT), às 72h e 96h, respectivamente. O magnésio se apresentou, no início do experimento (0h) e ao final (30d), significativamente maior nos grupos das búfalas (BT e BC). A laparotomia e ou o tratamento com fenilbutazona não interferiram nas concentrações de sódio, potássio e cálcio ionizado / Abstract: This study aims to compare the effect of exploratory laparotomy and treated with phenylbutazone in bovine and buffaloes. To pursue this purpose, biochemical profile, proteinogram and blood count cell tests were performed in 28 animals, of which 14 were buffaloes and 14 cows, all of then non-nursing, distributed in four groups - VT: cows treated with phenylbutazone; VC: control cows; BT: buffaloes treated with phenylbutazone and BC: control buffaloes. Animals from VT and BT received a single administration of 10 mg/kg of phenylbutazone, intravenously one hour before surgery. Blood samples were taken before laparotomy (0h) and 1(1h), 6(6h), 12(12h), 24(24h), 48(48h), 72(72h), 96(96h) e 120 horas (120h) e 15(15d) e 30(30d) days after surgery. Creatinine values were higher in buffaloes, and there were no changes, despite surgery or treatment. HDL showed different values in both species, due to surgical procedure. The use of phenylbutazone increased serum HDL in cows and buffaloes. Among acute phase proteins, serum haptoglobin showed higher values in buffaloes than in cows. Hemopexina was foundetected only in buffaloes and until the 5th day. Treatment with phenylbutazone did not change RBCs or globular volume values. Cows and buffaloes showed neutrophilia 12 hours after laparotomy. Phosphorus content increased in control buffaloes (BC) and in the treated buffaloes (BT) at 72h and 96h, respectively. Magnesium presented higher itself at the beginning of the experiment (0h) and end (30d) in buffaloes groups (BT and BC). Laparotomy or treatment with phenylbutazone did not changed sodium, potassium and calcium ionized values / Doutor

Haptoglobin is an acute phase protein with important biological role because of its capacity to bind to haemoglobin. Haptoglobin exists in three major genetic polymorphism types: Hp1-1, Hp2-1 and Hp2-2, the distribution of which has been associated with abdominal aortic aneurysm (AAA), an asymptomatic aortic disease common among men older than 65 years.    Five different electrophoretic methods were tested according to their ability to separate the haptoglobin phenotypes. The detection was based on a produced hemolysate of blood in which haemoglobin binds to haptoglobin thereby forming a complex that can be detected by specific haemoglobin staining using TMB-dihydrochloride and hydro peroxide as substrate resulting in an azure-green color of the bands. Samples from 15 patients who had suffered surgery for not broken AAA, that is more than5.0 cmaortic diameter, and 15 samples from matched controls were analyzed.    Among the five tested electrophoretic methods best migration and separation was seen on the pre-cast agarosgel Hydragel HR on the instrument Hydrasys. The other four methods gave less successful results. This pilot investigation showed the following distribution of the phenotypes of haptoglobin among AAA patients; 7 % Hp1-1, 40 % Hp2-1 and 53 % Hp2-2 and for the controls; 13 % Hp1-1, 33 % Hp2-1 and 53 % Hp2-2.    In conclusion, the used techniques has to be further optimized and more patients have to be included in the study before it can be ascertained if the phenotypes of Haptoglobin play any role in the progress of the AAA disease.

Orientador: Marco Antonio de Andrade Belo / Resumo: O presente estudo investigou os efeitos da ciclofosfamida (CYP) na modulação de proteínas de fase aguda (APPs) e acúmulo celular em exsudatos presentes na bexiga natatória de tilápias (modelo de peixe de aerocistite) e o recrutamento de leucócitos na lamínula de vidro implantadas no tecido subcutâneo. Foram utilizadas 140 tilápias (Oreochromis niloticus) (± 150 g) distribuídas em 14 aquários com capacidade para 250 L de água (n = 10). Os animais foram distribuídos em dois ensaios: Experimento I: foram utilizados 80 animais distribuídos em quatro tratamentos: T1 - controle NAIVE; T2 - controle negativo + solução salina injetada NaCl; T3- controle positivo + injetado com Aeromonas hydrophila; T4- ciclofosfamida, Isopac ® Sigma (200 mg / kg, dose única, por via intraperitoneal) + injetado com A. hydrophila. As amostras de exsudato e sangue foram coletadas 6 e 24 horas após a infecção (HPI) para a contagem de células e determinação do fracionamento eletroforético das APPs por SDS-PAGE, digestão de proteínas em gel e identificação por espectrometria de massa. Experimento II: os peixes foram alocados (n=60) em 3 grupos experimentais: G1 - controle NAIVE; G2 – controle positivo + implante de lamínula; G3 - ciclofosfamida, Isopac ® Sigma (200 mg / kg, dose única, por via intraperitoneal) + implante de lamínula. Após serem anestesiados, foram submetidos ao implante da lamínula de vidro de 10mm de diâmetro no tecido subcutâneo. As amostras de sangue foram coletadas e as lamínulas foram... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The present study investigated the effects of cyclophosphamide (CYP) on the modulation of acute phase proteins (APPs) and cellular accumulation in exudates present in the swim bladder of tilapia (aerocystitis fish model) and the recruitment of leukocytes on a glass cover slip implanted in the subcutaneous tissue. One hundred forty tilapia (Oreochromis niloticus) (± 150 g) were distributed in 14 aquariums with capacity of 250 L (n = 10), for two trials. Experiment I: For this purpose, 80 fish were distributed in four treatments: T1 – naïve control; T2 - negative control + injected with NaCl solution; T3- positive control + injected with A. hydrophila; and T4- cyclophosphamide (200 mg/kg) + injected with A. hydrophila. Samples of exudate and blood were collected 6 and 24 hours post-inoculation (HPI) for cell counts and determination of the APP electrophoretic fractionation by SDS-PAGE, in-gel protein digestion and mass spectrometric identification. Experiment II: Fish were allocated (n = 60) in 3 experimental groups: G1 – naive control; G2 - positive control + coverlet implant; and G3 - cyclophosphamide (200 mg/kg, single dose intraperitoneally) + cover slip implant. The glass coverslips (10 mm diameter) were implanted after anesthetization in the subcutaneous tissue. Blood samples were collected and the coverslips were carefully removed and washed with 0.9% saline 74 and 144 hours implantation. Then they were fixed in Bouin's solution and stained with hematoxylineosin. The tot... (Complete abstract click electronic access below) / Doutor

O presente estudo avaliou a dinâmica da onfalite, diarreia e Doença Respiratória Bovina (DRB), além do comportamento da haptoglobina (Hp) em bezerras Holandesas nos primeiros 41 dias de vida. Ainda, foi possível determinar os parâmetros de performance da Hp frente à detecção de doenças. Para tanto, foram avaliadas 216 bezerras nos momentos D2-6; D7-13; D14-20; D21-27 e D31-41. A haptoglobina (Hp) foi mensurada por técnica espectrofotométrica, enquanto a sanidade das bezerras foi avaliada segundo o Calf Health Scoring Criteria, da University of Wisconsin Madison. A diarreia foi monitorada pela avaliação da consistência das fezes, sendo adotada a seguinte classificação: escore 0 fezes com consistência normal; escore 1- fezes pastosas e semi-formada; escore 2- fezes pastosas com maior quantidade de água e conteúdo fecal aderido ao períneo e cauda; escore 3- fezes líquidas com conteúdo fecal aderido no períneo e cauda. Escores 2 e 3 foram considerados positivos para diarreia. A matéria seca das fezes também foi determinada para avaliação da saúde intestinal, sendo os animais com valores ≤15% classificados como positivos para diarreia. A DRB foi determinada adotando-se os seguintes parâmetros: tosse, secreção nasal, secreção ocular, posicionamento de orelhas e temperatura corpórea, sendo pontuados de 0 a 3 de acordo com a intensidade da sintomatologia. Soma das pontuações escore ≥5 foi adotada para classificar os animais positivos para DRB. As onfalites foram determinadas pela inspeção e palpação das estruturas umbilicais externas. O período de maior prevalência da onfalite foi no D7-13 (8,8%; 19/216), seguida pelas diarreias no D14-20 (42,9%; 76/177) e DRB no D31-41 (26,4%; 55/208). A média da concentração de Hp durante o experimento foi de 0,03±0,05 g/L, com valores mínimo e máximo de 0,00 e 0,52 g/L, respectivamente. Os picos de haptoglobina foram detectados nos momentos D7-13 (0,04±0,07) e D14-20 (0,04±0,05 g/L). Em relação às diarreias (escore fecal 2 e 3), a Hp apresentou sensibilidade de 56,9% e especificidade 74,07%, sendo o ponto de corte estabelecido equivalente a 0,02 g/L. Resultados semelhantes foram observados para diarreias determinadas pelo teor de matéria seca: sensibilidade 58,1; especificidade 76,2; e ponto de corte 0,02 g/L. Em relação a DRB, a sensibilidade e especificidade foi de 72,7 e 81,2%, respectivamente, quando utilizado a soma do escore respiratório≥ 5 associado à temperatura retal >39,5ºC, com ponto de corte de 0,02 g/L. O ciclo de doenças foi caracterizado cronologicamente pelas onfalites, diarreias e DRB nos primeiros 41 dias de vida. A haptoglobina pode ser usada como biomarcador para a detecção da DRB. Por outro lado, a proteína apresentou baixa sensibilidade para detecção de diarreia usando dois padrões-ouro. / This research evaluated the dynamic of navel inflammation, diarrhea and Bovine Respiratory Disease (BRD), besides the haptoglobin (Hp) profile in Holstein heifers in the first 41 days of life. Moreover, it was possible to determine the development of haptoglobin to detect diseases. For this, it was evaluated 216 Holstein heifers in the following moments D2-6; D7-13; D14-20; D21-27 e D31-41. Haptoglobin was measured using spectrophotometric, and the health status of calves were assessed in accordance with the Calf Health Scoring Criteria from The University of Wisconsin (Madison). Diarrhea was monitored according to fecal consistency, using the following classification: score 0- normal consistency, score 1- pasty, semi-formed, score 2- pasty with largest amount of water, score 3- liquid with fecal content adhered in the perineum and tail. Calves were assessed having diarrhea when the score were 2 or 3. Fecal dry matter also was determined to evaluate the gut health, so animals with values ≤15% were classified such as positive for diarrhea. BRD was assessed using the following parameters: rectal temperature, nasal secretion, ocular secretion, cough and ear/head position with score of 0-3 based on severity of each. Calves were assessed having BRD when the sum of these scores was ≥5. Omphalitis was evaluated by inspection and palpation of navel external structures. The highest prevalence of omphalitis was observed on D7-D13 (8.8%; 19/216), followed diarrhea on D14-D20 (42.9%, 76/177) and BRD on D31-D41 (26.4%; 55/208). The mean of Hp along this research was 0.03±0.05 g/L, with minimum and maximum values of 0.00 e 0.52 g/L, respectively. The peak of haptoglobin was detected on D7-13 (0.04±0.07) e D14-20 (0.04±0.05 g/L). In relation to diarrhea (fecal score 2 and 3), the haptoglobin had sensibility of 56.9% and specificity of 74.07%, and the cut-off established was 0.02 g/L. Similar results was observed for diarrhea determined using fecal dry matter: sensibility of 58.1; specificity of 76.2; and cut-off of 0.02 g/L. In relation to BRD, sensibility and specificity detected was 72.7 and 81.2%, respectively, when the sum of score was ≥5 associated with rectal temperature >39.5ºC, with cut-off equal 0.02 g/L. The cycle of diseases was characterized by omphalitis, diarrhea and BRD in the first 41 days of life. Haptoglobin can be used as a biomarker to detect BRD. On the other hand, this protein had low sensibility to detect diarrhea using two gold standard.

Lipoprotein(a) (Lp(a)) consists of an LDL-like particle and the specific protein apo(a), which is very similar to plasminogen. Apo(a) contains repeated kringle structures and a serine protease domain, which cannot be activated by t-PA. Lp(a) is considered to be a predictor for atherosclerotic disease. It has been found incorporated in atherosclerotic plaques and inhibits in vitro fibrinolysis. Lp(a) was determined in 1527 randomly selected individuals participating in the Northern Sweden WHO-MONICA project. A weak but significant relation between Lp(a) and increasing age was found. Menopausal status was the strongest independent predictor of Lp(a) level in women. Fibrinogen was independently related to Lp(a) in both sexes. Only a minor fraction of Lp(a) variance could be explained for in a multiple regression model, which is in agreement with the contention that Lp(a) is highly genetically determined. Lp(a) was determined in 1571 patients investigated with coronary angiography because of suspected severe coronary artery disease (CAD). Patients with proven CAD at elective angiography had significantly higher Lp(a) than patients without significant CAD or healthy controls. Lp(a) was found to be an independent discriminator of CAD in both sexes. HLA-DR genotype 13 or 17 was found more frequently in 30 male patients with angiographic CAD at young age (&lt; 50 years) than in 30 age matched controls. These genotypes were common in patients with high Lp(a) levels, which indicates that Lp(a) may be related to immunological processes. The reaction of Lp(a) was investigated in 32 patients with acute myocardial infarction (AMI). Lp(a) increased during the first week, but the response was comparatively weak. Individual Lp(a) responses were heterogeneous and no correlations to infarct size or changes in the acute phase proteins were found. In a randomized cross-over study on 36 hypercholesterolaemic patients treated with simvastatin/placebo during 12+12 weeks Lp(a) did not change significantly, but patients with high Lp(a) levels at baseline tended to develop further increased Lp(a). To conclude, Lp(a) was found to be an independent predictor of angiographic CAD in both men and women. Lp(a) levels are primarily genetically determined and only a small fraction of Lp(a) variance could be explained by other factors in this study. Lp(a) may be related to HLA DR types and immunological processes involved in atherosclerotic disease. Lp(a) increased slightly during the first week of AMI, but was not related to changes in the acute-phase proteins. The effective LDL-lowering agent simvastatin did not influence Lp(a) significantly. / <p>Diss. (sammanfattning) Umeå : Umeå universitet, 1993, härtill 5 uppsatser.</p> / digitalisering@umu

Die Bestimmung der Plasmaviskositaet und des Haptoglobin beim Rind erweisen sich als sinnvolle Ergaenzungen in der Labordiagnostik. Ihre Bestimmung als Suchtest zur Fruehdiagnostik von praxisrelevanten Rinderkrankheiten ist zu empfehlen.

Die Bestimmung der Plasmaviskositaet und des Haptoglobin beim Rind erweisen sich als sinnvolle Ergaenzungen in der Labordiagnostik. Ihre Bestimmung als Suchtest zur Fruehdiagnostik von praxisrelevanten Rinderkrankheiten ist zu empfehlen.

C-reactive protein (CRP) is synthesized in hepatocytes. The serum concentration of CRP increases dramatically during the acute phase response. In human hepatoma Hep3B cells, maximal CRP expression occurs in cells treated with the combination of IL-6 and IL-1β. IL-6 induces transcription of the CRP gene and IL-1β synergistically enhances the effects of IL-6. We investigated the role of IL-6-activated transcription factor STAT3, also known as STAT3α, in inducing CRP expression since we identified four consensus STAT3-binding sites centered at positions - 72, - 108, - 134 and - 164 on the CRP promoter. It has been shown previously that STAT3 binds to the site at - 108 and induces CRP expression. We found that STAT3 also bound to the other three sites, and several STAT3-containing complexes were formed at each site, suggesting the presence of STAT3 isoforms and additional transcription factors in the complexes. Mutation of the STAT3 sites at - 108, - 134 or - 164 resulted in decreased CRP expression in response to IL-6 and IL-1β treatment, although the synergy between IL-6 and IL-1β was not affected by the mutations. The STAT3 site at - 72 could not be investigated employing mutagenesis. We also found that IL-6 activated two isoforms of STAT3 in Hep3B cells: STAT3α which contains both a DNA-binding domain and a transactivation domain and STAT3β which contains only the DNA-binding domain. Taken together, these findings raise the possibility that IL-6 not only induces CRP expression but also regulates the induction of CRP expression by activating STAT3 isoforms and by utilizing all four STAT3 sites.