<p>Tokom perioda rane laktacije brojni unutra&scaron;nji i<br />spolja&scaron;nji faktori vr&scaron;e uticaj na produktivnost i<br />zdravlje mlečnih krava. Najznačajniji unutra&scaron;nji<br />faktori na početku laktacije su zapaljenski<br />procesi i metabolički stres. S obzirom da su<br />metaboličke i imunolo&scaron;ke promene najče&scaron;ći<br />okidač za nastanak bolesti, neophodno je bilo<br />ispitati da li postoji mogućnost regulisanja ovih<br />promena pomoću nesterodinih antiinflamatornih<br />lekova. Cilj ove doktorske disertacije je bio da<br />se ispita uticaj parenteralne primene<br />ketoprofena na metabolički status, inflamatorne<br />promene, odnos između metaboličkih i<br />inflamatornih promena kod krava posle teljenja,<br />kao i na proizvodnju mleka kod krava u ranoj<br />laktaciji. Za ogled je odabrano 30 krava.<br />Oglednoj grupi od 15 krava je i.m. aplikovan<br />ketoprofen u terapijskoj dozi (3mg/kg telesne<br />mase) tokom tri uzastopna dana nakon teljenja.<br />Kontrolna grupa od 15 krava je predstavljala<br />negativnu kontrolu. Uzorci krvi za analizu su<br />uzimani iz repne vene (vena coccygea) na dan<br />teljenja, u prvoj i drugoj nedelji posle teljenja.<br />Procena inflamacije je vr&scaron;ena na osnovu<br />određivanja koncentracije proteina akutne faze<br />(haptoglobina i fibrinogena) i proinflamatonih<br />citokina (interleukina-1&alpha;, faktora nekroze<br />tumora-alfa i interferona gama), dok je procenametaboličkog statusa vr&scaron;ena na osnovu određivanja vrednosti metaboličkih parametarau svim uzorcima krvnog seruma i u svim nedeljama ogleda. Ketoprofen primenjivan parenteralno kod krava posle teljenja je ublažio metabolički stres, uticao na smanjenje inflamatornih procesa i na smanjnje međusobne povezanosti i uticaja između metaboličkih i inflamatornih promena kod krava posle teljenja u poređenju sa netretiranim kravama, a imao je i pozitivan uticaj na proizvodnju mleka u ranoj laktaciji.</p> / <p>During the period of early lactation numerous<br />internal and external factors influence the health<br />and productivity of dairy cows. The most<br />important internal factors at the beginning of<br />lactation are inflammatory processes and<br />metabolic stress. Since the metabolic and<br />immunologic changes are the most common<br />trigger for the disease, it was necessary to<br />examine whether there is a possibility of<br />regulating these changes by using non-steroidal<br />anti-inflammatory drugs. The aim of this<br />doctoral thesis was to investigate the effect of<br />parenteral administration of ketoprofen in the<br />metabolic status, inflammatory changes, the<br />relationship between metabolic and<br />inflammatory changes in cows after calving and<br />milk production in cows in early lactation. This<br />study included 30 cows. Experimental group of<br />15 cows is treated i.m. ketoprofen administered<br />in a therapeutic dose (3 mg / kg body weight)<br />for three consecutive days after calving. A<br />control group of 15 cows accounted for a<br />negative control. Blood samples for analysis<br />were taken from the tail vein (vena coccygea)<br />on the day of calving, in the first and second<br />week after calving. Evaluation of inflammation<br />was based on the determination of the<br />concentration of acute phase proteins<br />(haptoglobin and fibrinogen) and<br />proinflamatonih cytokines (interleukin-1&alpha;,tumor necrosis factor-alpha and interferongamma),and the evaluation of the metabolic status performed on the basis of determining the value of metabolic parameters in a blood sample serum and in all the weeks of the experiment. Ketoprofen administered parenterally in cows after calving eased metabolic stress, had impact on reducing inflammatory processes and the decrease of interconnection and influence between metabolic and inflammatory changes in cows after calving compared with untreated cows, and had a positive impact on milk production in the early lactation.</p><p>&nbsp;</p>

A distrofia muscular de Duchenne (DMD) é uma alteração neuromuscular hereditária e progressiva que afeta humanos do sexo masculino. O modelo canino Golden Retriever Muscular Dystrophy (GRMD) é considerado modelo experimental para estudos de novas propostas terapêuticas e melhor entendimento da fisiopatogênica da DMD. O processo progressivo da distrofia está relacionado com alterações nas populações celulares que compõe o sistema imune dos pacientes, pois devido a ausência da proteína distrofina na membrana sarcoplasmática, o músculo fica mais susceptível à lesões, ocorrendo liberação de citocinas, que recrutam e estimulam células do sistema imune, principalmente macrófagos e linfócitos T. A longo prazo, essa resposta inflamatória contínua e persistente, leva a uma série de reações que culminam com danos cada vez maiores, a ponto de ocorrer um esgotamento de células satélites e fibrose do tecido muscular. Uma das propriedades mais estudadas das células tronco mesenquimais (MSCs) é sua capacidade imunomoduladora, fazendo com que essas células se tornem promissoras na utilização da terapia celular. Neste contexto, esta ferramenta imunomoduladora pode atuar como uma estratégia interessante na manipulação do sistema imune. O estudo proposto foi elaborado com a finalidade de trazer subsídios para viabilização de experimentos de terapia celular na distrofia muscular, por meio do conhecimento sobre a imunomodulação durante o tratamento in vitro, contribuindo para uma possível aplicação terapêutica em humanos. Para tanto, foram estudados dois grupos de cães, um grupo controle (GR; n=5) e um grupo de cães afetados (GRMD; n=9), compostos por machos e fêmeas. O estudo consistiu na avalição da proliferação de linfócitos na presença de MSC em diferentes concentrações, bem como da proliferaçao de linfócitos específicos como o Tauxiliar (CD4+FoxP3-) e Tregulatórios (CD4+FoxP3+) com as MSCs. Adicionalmente, realizamos a dosagem de nitrito com o intuito de quantificar a produção de óxido nítrico (NO) através do cocultivo de macrófagos com as MSCs. Neste estudo foi possível observar que as MSCs estimularam a proliferação significativa de linfócitos T regulatórios. Adicionalmente, essa porcentagem de divisão aumentou em cocultivos que utilizaram maiores concentrações de MSC. Maior concentração de nitrito também foi encontrada no cocultivos de MSC e macrófagos estimulado com LPS. Estas informações geram incrementos no entendimento de como as MSCs podem agir no organismo distrófico e como poderemos explorar essa fonte para promover um retardamento no processo inflamatório e consequentemente melhorar a qualidade de vida do paciente. / The Duchenne muscular dystrophy (DMD) is a progressive hereditary neuromuscular disorder that affects human males. The canine model Golden Retriever Muscular Dystrophy (GRMD) is considered experimental model for studies of new therapies and better understanding of the DMD. The process of progressive dystrophy is related to changes in cell populations that comprise the immune system of patients, because due to the absence of the protein dystrophin in the sarcoplasmic membrane, the muscle is more susceptible to injury, occurring release of cytokines that recruit and stimulate cell immune, mainly macrophages and T lymphocytes in the long term, this continuous and persistent inflammatory response, the system takes a series of reactions that culminate with increasing damage to the point of exhaustion of satellite cells of muscle tissue and fibrosis occur. One of the most studied properties of mesenchymal stem cells (MSCs) is their immunomodulatory capacity, making these cells become promising in the use of cell therapy. In this context, this immunomodulatory can act as an interesting strategy in manipulating the immune system. The proposed study was designed in order to provide support for the feasibility of cell therapy trials in muscular dystrophy, through the knowledge of immunomodulation during treatment in vitro, contributing to a possible therapeutic application in humans. In this purpose, two groups were studied, a group of affected dog (GRMD, n=9) and a control group (GR, n=5 GR). The study consisted of rating of lymphocyte proliferation in the presence of MSCs in different concentrations as well as the proliferation of specific lymphocytes as Thelper (CD4+FoxP3-) and Tregulatory (CD4+FoxP3+) to MSCs. In addition, the dosage of nitrite performed in order to quantify the production of nitric oxide (NO) by coculture with macrophages MSCs. In this study we observed that MSCs stimulated significant proliferation of regulatory T lymphocytes. Additionally, this percentage split increased cocultivos that used higher concentrations of MSC. Highest concentration of nitrite was also found in cocultivos of MSC and macrophages stimulated with LPS. This information generates increments in understanding how MSCs may act in the body dystrophic and how we exploit this source to promote a delay in the inflammatory process and consequently improve the quality of life of patients

Embora camundongos sejam resistentes à infecção por Leptospira interrogans, eles têm sido pouco utilizados para se entender os mecanismos imunes efetores contra esta bactéria. Adler & Faine mostraram em 1976 que esta resistência é dependente da resposta imune, uma vez que camundongos imunossuprimidos tornavam-se suscetíveis à L. interrogans. Outros autores mostraram que camundongos portadores de imunodeficiência grave combinada também morriam após inoculação de L. interrogans. Sabendo da importância do Sistema Complemento em infecções bacterianas, investigamos se animais C5 deficientes (C5-) são mais suscetíveis à infecção por L. interrogans que animais C5 normais (C5+). Observamos que camundongos C5- possuem menores porcentagens de linfócitos T CD8+ na circulação periférica e maiores níveis de IL-12p40 no rim e de TNF e IL-6 no pulmão que os animais C5+. Animais C57Bl/6 (B6) C5+ possuem maior porcentagem de linfócitos T CD4+ que B6 C5-, além de lesões hepáticas mais intensas, mostrando um efeito dependente de C5 e do fundo genético dos camundongos. / Although mice are resistant to Leptospira interrogans infection, they are not usual models to study the imune response against this bacteria. Adler and Faine demonstrated in 1976 the importance of the imune response, since immunosuppressed mice were suceptible to L. interrogans. Other authors showed that mice that had severe combined immunodeficiency also died when inoculated with L. interrogans. Due to the activity of the Complement System in infections, we analyzed whether C5 deficient (C5-) mice are more susceptible than C5 sufficient (C5+) mice to L. interrogans infection. C5- mice have lower percentages of T CD8+ lymphocytes in peripheral circulation and upper levels of IL-12p40 in the kidney and TNF and IL-6 in the lungs than C5+ mice. C57Bl/6 (B6) C5+ mice has higher percentages of T CD4+ lymphocytes than B6 C5- mice, in addition to stricter liver injures, exhibiting an effect dependent of C5 and of the genetic background.

Embora camundongos sejam resistentes à infecção por Leptospira interrogans, eles têm sido pouco utilizados para se entender os mecanismos imunes efetores contra esta bactéria. Adler & Faine mostraram em 1976 que esta resistência é dependente da resposta imune, uma vez que camundongos imunossuprimidos tornavam-se suscetíveis à L. interrogans. Outros autores mostraram que camundongos portadores de imunodeficiência grave combinada também morriam após inoculação de L. interrogans. Sabendo da importância do Sistema Complemento em infecções bacterianas, investigamos se animais C5 deficientes (C5-) são mais suscetíveis à infecção por L. interrogans que animais C5 normais (C5+). Observamos que camundongos C5- possuem menores porcentagens de linfócitos T CD8+ na circulação periférica e maiores níveis de IL-12p40 no rim e de TNF e IL-6 no pulmão que os animais C5+. Animais C57Bl/6 (B6) C5+ possuem maior porcentagem de linfócitos T CD4+ que B6 C5-, além de lesões hepáticas mais intensas, mostrando um efeito dependente de C5 e do fundo genético dos camundongos. / Although mice are resistant to Leptospira interrogans infection, they are not usual models to study the imune response against this bacteria. Adler and Faine demonstrated in 1976 the importance of the imune response, since immunosuppressed mice were suceptible to L. interrogans. Other authors showed that mice that had severe combined immunodeficiency also died when inoculated with L. interrogans. Due to the activity of the Complement System in infections, we analyzed whether C5 deficient (C5-) mice are more susceptible than C5 sufficient (C5+) mice to L. interrogans infection. C5- mice have lower percentages of T CD8+ lymphocytes in peripheral circulation and upper levels of IL-12p40 in the kidney and TNF and IL-6 in the lungs than C5+ mice. C57Bl/6 (B6) C5+ mice has higher percentages of T CD4+ lymphocytes than B6 C5- mice, in addition to stricter liver injures, exhibiting an effect dependent of C5 and of the genetic background.

Coronary heart disease is the leading cause of death in American women, accounting for one in six deaths in 2004. One third of women over the age of forty will develop coronary heart disease in their lifetime. The role of chronic and excessive inflammation and risk factors, such as smoking and high cholesterol, are now well-established factors contributing to coronary heart disease pathology. A knowledge gap exists in that little is known about the mechanisms by which psychosocial factors, such as anger, may be associated with pro-inflammatory processes that contribute to cardiovascular morbidity and mortality in women. The purpose of this study was to determine if there were differences in serum levels of the proinflammatory biomarker, C-reactive protein, in post-menopausal women who scored high on anger characteristics compared to those with low anger characteristics. Mean levels of C-reactive protein were not found to be different in a sample of 42 women with high trait anger or high anger expression compared to those with low trait anger or low anger expression. Significant relationships were found in C-reactive protein and some anger control characteristics (anger control-in) and might imply that certain anger expression styles may play a role in pro-inflammatory responses in post-menopausal women.

Biologically inspired materials are being developed with the aim of improving the integration of medical implants and minimizing non-desirable host reactions. A promising strategy is the design of topographically patterned surfaces that resemble those found in the extracellular environment. Nanoporous alumina has been recognized as a potential biomaterial and as an important template for the fabrication of nanostructures. In this thesis in vitro studies were done to elucidate the role of alumina nanoporosity on the inflammatory response. Specifically, by comparing alumina membranes with two pore sizes (20 and 200 nm in diameter). Complement and platelet activation were evaluated as well as monocyte/macrophage behaviour. Whole blood was incubated with the alumina membranes and thereafter the biomaterial surfaces were evaluated in terms of protein and platelet adhesion as well as procoagulant properties. The fluid phase was analyzed for complement activation products and platelet activation markers. Besides, human mononuclear cells were cultured on the alumina membranes and cell adhesion, viability, morphology and release of pro-inflammatory cytokines were evaluated. The results indicated that nanoporous alumina with 200 nm pores promotes higher complement activation than alumina with 20 nm pores. In addition, platelet response to nanoporous alumina was found to be highly dependent on the material porosity, as reflected by differences in adhesion, PMP generation and procoagulant characteristics. A clear difference in monocyte/macrophage adhesion and activation was found between the two pore size alumina membranes. Few but highly activated cells adhered to the 200 nm membrane in contrast to many but less activated monocytes/macrophages on the 20 nm surface. The outcome of this work emphasizes that nanotopography plays an important role in the host response to biomaterials. Better understanding of molecular interactions on nano-level will undoubtedly play a significant role in biomaterial implant development and will contribute to design strategies for controlling specific biological events.

ABSTRACT In the modern era, the most common post-operative complications following cardiopulmonary bypass (CPB) are neurocognitive deficits (NCD) and atrial fibrillation (AF). Both morbidities have been linked to inflammation resulting from surgery, anesthesia and CPB. Microemboli, inadequate oxygen delivery and the inflammatory response consequent to blood contacting artificial components of the CPB circuit have all been linked to postoperative NCD and to a lesser extent post-operative AF. The artificial components of the CPB circuit consist of stainless steel, polyvinylchloride (PVC), polycarbonate and other carbon-based plastics. In order to attenuate the negative sequelae of blood-circuit contact related inflammatory response, industry developed the biocompatible circuit (BCC) coating for the disposable CPB circuits. Four such coatings were studied and compared to an uncoated control group in a total of 101 patients undergoing routine CPB-assisted cardiac surgical procedures. Soluble adhesion molecule (SAM) activation was studied at different time points and common clinical outcomes such as white blood cell activation, serum renal function parameters urea and creatinine, postoperative bleeding, transfusion requirements, intensive care and hospital length of stay, CPB pump volume balances, changes in weight, postoperative serum lactate and glucose and the development of AF postoperatively, were compared. Additionally, postoperative neurocognitive testing was performed using a simple bedside neurocognitive test called the antisaccadic eye movement test. The patients in all groups were tested for comparison preoperatively and 72 hr postoperatively. Results: The mandate of BCC coating development and manufacture is to attenuate the well-documented and demonstrated inflammatory response consequent to the contact of blood with artificial CPB surfaces. The studied BCCs significantly decreased platelet transfusions in females. In addition, the BCCs decreased the concentrations of 2 SAMs when measured 6 hours after surgery and CPB. The difference in SAM expression seen between the coated and uncoated groups at 6 hr was no longer apparent at 72 hr. Very little difference was noted between the four BCC groups. Patients who developed AF postoperatively seemed predisposed to do so as the serum levels of soluble vascular cell adhesion molecule was significantly higher at baseline and remained so at 6 and 72 hr. The decreased platelet transfusions in females resulting from BCC use is a highly significant finding within this high-risk group of patients. As most platelet transfusions occur soon after the patient is disconnected from CPB, the short-term decrease in SAM activation can be linked to this improved clinical finding. The studied BCC coatings have achieved limited success in their intended mandate to attenuate inflammatory response in terms of improved clinical and laboratory desired outcomes.

ABSTRACT In the modern era, the most common post-operative complications following cardiopulmonary bypass (CPB) are neurocognitive deficits (NCD) and atrial fibrillation (AF). Both morbidities have been linked to inflammation resulting from surgery, anesthesia and CPB. Microemboli, inadequate oxygen delivery and the inflammatory response consequent to blood contacting artificial components of the CPB circuit have all been linked to postoperative NCD and to a lesser extent post-operative AF. The artificial components of the CPB circuit consist of stainless steel, polyvinylchloride (PVC), polycarbonate and other carbon-based plastics. In order to attenuate the negative sequelae of blood-circuit contact related inflammatory response, industry developed the biocompatible circuit (BCC) coating for the disposable CPB circuits. Four such coatings were studied and compared to an uncoated control group in a total of 101 patients undergoing routine CPB-assisted cardiac surgical procedures. Soluble adhesion molecule (SAM) activation was studied at different time points and common clinical outcomes such as white blood cell activation, serum renal function parameters urea and creatinine, postoperative bleeding, transfusion requirements, intensive care and hospital length of stay, CPB pump volume balances, changes in weight, postoperative serum lactate and glucose and the development of AF postoperatively, were compared. Additionally, postoperative neurocognitive testing was performed using a simple bedside neurocognitive test called the antisaccadic eye movement test. The patients in all groups were tested for comparison preoperatively and 72 hr postoperatively. Results: The mandate of BCC coating development and manufacture is to attenuate the well-documented and demonstrated inflammatory response consequent to the contact of blood with artificial CPB surfaces. The studied BCCs significantly decreased platelet transfusions in females. In addition, the BCCs decreased the concentrations of 2 SAMs when measured 6 hours after surgery and CPB. The difference in SAM expression seen between the coated and uncoated groups at 6 hr was no longer apparent at 72 hr. Very little difference was noted between the four BCC groups. Patients who developed AF postoperatively seemed predisposed to do so as the serum levels of soluble vascular cell adhesion molecule was significantly higher at baseline and remained so at 6 and 72 hr. The decreased platelet transfusions in females resulting from BCC use is a highly significant finding within this high-risk group of patients. As most platelet transfusions occur soon after the patient is disconnected from CPB, the short-term decrease in SAM activation can be linked to this improved clinical finding. The studied BCC coatings have achieved limited success in their intended mandate to attenuate inflammatory response in terms of improved clinical and laboratory desired outcomes.

Purpose: Traumatic brain injury (TBI) leads to important and deleterious neuroinflammation, as evidenced by edema, cytokine production, induction of nitric oxide synthase, and leukocyte infiltration. Strategies that block inflammatory and oxidative mediators have been shown to induce neuroprotective and anti-inflammatory effects after brain injury. After TBI, cerebral vascular endothelial cells play a crucial role in the pathogenesis of inflammation. In this study, we hypothesized that cerebral vascular endothelial cells play a crucial role in the pathogenesis of inflammation after TBI and, in conjunction with leukocytes, represent a key cellular target for statin therapy. We investigated the effect of acute and continuous treatment of simvastatin on behavior and inflammation in adult rats following experimental TBI. Materials and Methods: Cortical contusions were induced using a device adapted from the impact method. There were 3 groups: (1) sham group, craniotomy only; (2) control group, TBI without treatment; and (3) treatment group, TBI with simvastatin administration. The treatment group received 15 mg/kg of simvastatin daily for 3 days. Neurological function was assessed with the grip test (Grip strength meter, Singa). Results: Non-treatment control group had a significantly greater increase in ICAM-1 expression from pre-injury to the post-injury 72 h time point, compared to the simvastatin treatment group. The treatment group had a significantly smaller amount of reduction in successful trials in grip test than the control group did from baseline to 72 h. The analysis of western blot and pathological study also demonstrated similar results. Conclusion: Our findings indicate that continuous administration of simvastatin after injury attenuates the cerebral vascular endothelial inflammatory response and improves functional and histological outcomes in a rat model of TBI. This improvement is associated with a reduction in expression of ICAM-1 in the blood and brain after rat TBI when compared with the untreated control group. Hence, we recommend simvastatin administration in the first 72 h following TBI.

Early detection of infection, identification of microorganism, and correct choice of antibiotics are critical in the management of sepsis. Quantitative real-time polymerase chain reaction (RT-PCR) has the potential to improve the timeliness, sensitivity, and accuracy of detecting pathogens. In this study we utilize this method to detect the enteric bacterial counts in the blood from patients with systemic inflammatory response syndrome (SIRS) in the emergency department (ED). The universal primers utilized in RT-PCR are specific for 23S ribosomal DNA (rDNA) and wec F gene. The results show that in SIRS patients with positive culture results from specimen collected within 10 days after presenting to ED, and patients surviving for less than 28 days, the serum bacterial DNA load of enteric Gram negative bacilli is higher. In SIRS patients with shock, patients fulfilling both white blood cell counts and respiratory criteria of SIRS, and patients fulfilling both white blood cell counts and respiratory criteria of SIRS with Acute Physiology and Chronic Health Evaluation II score more than 20, the serum bacterial DNA load of enteric Gram negative bacilli and 28-day mortality are both higher. These results suggest that bacterial translocation may happen in patients with SIRS and may be related to higher mortality in patients with SIRS.