Orientador: Miguel Gozalo Marcilla / Resumo: O objetivo principal do estudo foi avaliar os efeitos sedativos e cardiorrespiratórios da dexmedetomidina administrada pela via intravenosa em três diferentes doses em asininos Nordestinos. Na metodologia foram utilizadas seis jumentas, adultas (idade entre 3 e 5 anos), hígidas, com peso 132 kg ± 14 kg, provenientes da tropa da FMVZ- Unesp Botucatu. Os efeitos sedativos e comportamentais foram avaliados através da variação da altura da cabeça (HHAG), grau de ataxia, escala visual analógica (VAS) e respostas aos estímulos tátil (membro pélvico, torácico e orelha), auditivo e visual. Adicionalmente, foram analisadas as variáveis cardiorrespiratórias, frequência cardíaca (FC), pressão arterial sistólica (PAS) e frequência respiratória (FR), além de temperatura retal (Tº) e motilidade gastrointestinal (MGI). Cada animal recebia um dos quatro tratamentos que consistiam em: controle com solução fisiológica 0,9% (SAL), dexmedetomidina 3 μg/kg (D3), dexmedetomidina 5 μg/kg (D5) e dexmedetomidina 7 μg/kg (D7). As variáveis de sedação e cardiorrespiratórias foram registradas aos 5, 10, 20, 30, 45 e 60 minutos após a realização de cada tratamento. Dois avaliadores experientes também analisaram posteriormente a qualidade de sedação pela observação de registros em vídeo. Os dados normais foram analisados por medidas repetidas ANOVA sendo que as médias foram avaliadas pelo teste de Tukey (p > 0,05). Os testes de Friedman e de Dunn´s (p > 0,05) foram utilizados para a análise das variáveis ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The main objective of the study was to evaluate the sedative and cardiorespiratory effects of dexmedetomidine administered intravenously in three different doses in Northeastern asinines. In the methodology, six healthy adult donkeys weighing 132 kg ± 14 kg from the FMVZ-Unesp Botucatu troop were used. The sedative and behavioral effects were evaluated through head height variation (HHAG), ataxia degree, visual analog scale (VAS) and responses to tactile stimuli (pelvic, thoracic and ear member), auditory and visual. In addition, cardiorespiratory variables, heart rate (HR), systolic blood pressure (SBP) and respiratory rate (RR), as well as rectal temperature (Tº) and gastrointestinal motility (MGI) were analyzed. Each animal received one of the four treatments consisting of: control with physiological solution 0.9% (SAL), dexmedetomidine 3 μg / kg (D3), dexmedetomidine 5 μg / kg (D5) and dexmedetomidine 7 μg / kg (D7). Sedation and cardiorespiratory variables were recorded at 5, 10, 20, 30, 45 and 60 minutes after each treatment. Two experienced evaluators also subsequently analyzed the quality of sedation by watching video records. The normal data were analyzed by repeated measures ANOVA and the means were evaluated by the Tukey test (p> 0.05). The Friedman and Dunn's tests (p> 0.05) were used for the analysis of the variables that did not present normal distribution. The reduction in HHAG was significant in comparison to baseline, in all dexmedetomidine treatments (up to ... (Complete abstract click electronic access below) / Mestre

Submitted by SAMUEL DOS SANTOS SOUSA null (samuerr@hotmail.com) on 2016-11-25T13:56:01Z No. of bitstreams: 1 DOC. FINAL - DISSERTAÇÃO.pdf: 1281598 bytes, checksum: da30a95bebfc6a71081a5f812835a040 (MD5) / Approved for entry into archive by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-11-29T13:18:47Z (GMT) No. of bitstreams: 1 sousa_ss_me_jabo.pdf: 1281598 bytes, checksum: da30a95bebfc6a71081a5f812835a040 (MD5) / Made available in DSpace on 2016-11-29T13:18:47Z (GMT). No. of bitstreams: 1 sousa_ss_me_jabo.pdf: 1281598 bytes, checksum: da30a95bebfc6a71081a5f812835a040 (MD5) Previous issue date: 2016-08-18 / Existem poucos protocolos para a contenção e sedação de jumentos . Os agonistas alfa-2 são os fármacos mais amplamente administrados e é sabido que essa classe de fármacos pode produzir alguns efeitos deletérios no sistema cardiorrespiratório do animal. Os fármacos da classe dos opioides vêem ganhando espaço na prática anestésica com asininos, pois esses fármacos são utilizados como uma alternativa para a sedação desses animais. Além de produzirem certo grau de sedação, possuem característica analgésica, sem promover efeitos adversos. Na literatura pesquisada não foi encontrado, nenhum estudo sobre o uso associado de xilazina em infusão contínua com meperidina e seus efeitos hemodinâmicos nos muares. Diante deste exposto, objetivou-se estudar os efeitos da associação da infusão contínua da xilazina com bollus intramuscular de meperidina como protocolo de sedação em jumentos. Para tal, foram utilizados seis jumentos, SRD, sendo um macho e cinco fêmeas, com peso médio de 141±23 kg. Os animais foram submetidos a três protocolos experimentais dividos em três grupos, Grupo A: infusão contínua intravenosa de 1,1 mg/kg/hora de xilazina a solução salina por via intramuscular; Grupo B: infusão contínua intravenosa de 0,8 mg/kg/hora de xilazina e solução salina por via intramuscular e Grupo C: infusão contínua intravenosa de 0,8 mg/kg/hora de xilazina e 4 mg/kg de meperidina por via intramuscular. Foi observado redução na frequência respiratória, pressão arterial sistólica nos animais do grupo A e C, diminuição da pressão arterial diastólica e média em todos os grupos. Foi observada redução na altura da cabeça em todos os grupos. A associação da meperidina com xilazina em infusão contínua não provocou alterações cardiorrespiratórias significativas e produziu grau de sedação satisfatório / To the present, there has been few descriptions of anesthetic protocol for restraint and sedation in donkey (Equus asinus). Alpha-2-agonists are widely administered class of drugs. Known to produce some deleterious effects over the cardiopulmonary system. The use of opioids in donkeys has sained popularity anesthetic practicioners, since these drugs are used as an alternative sedative with analgesic characteristics with minimal side effects. To our knowledge, there has not been study evaluating the effects of the association of continuous rate infusion of xilazine with meperidine given intramuscularly over the cardiovascular parameters of donkeys. Therefore, the objective of our study evaluate was to the capacity of sedation and the cardiorespiratory implications of the anesthetic association in donkeys. In order to performe the study, six mixed breed, one male and five females, were used. The average weight has 141 ± 23 kg. The animals were subjected to three experimental protocols. Group A: Continuous intravenous infusion of 1.1 mg / kg / hour of xylazine and saline solution intramuscularly; Group B: Continuous intravenous infusion of 0.8 mg / kg / hour of xylazine and saline solution intramuscularly and Group C: continuous intravenous infusion of 0.8 mg / kg / hour of xylazine and 4 mg / kg of meperidine intramuscularly. Respiratory rate and systolic blood pressure decreased in animals of group A and C. While diastolic blood pressure and mean arterial pressure decreased in all three groups. Head Height lowered following treatment in all groups. The combination of meperidine with continuous rate infusion of xylazine did not cause significant cardiorespiratory implications and produced satisfactory degree of sedation.

Orientador: Paulo Sergio Patto dos Santos / Resumo: Objetivou-se com este trabalho avaliar os efeitos na hemodinâmica, respiração, motilidade ruminal e sedação, da associação de detomidina e nalbufina em ovinos. Foram utilizados 8 ovinos hígidos, jovens, fêmeas ou machos, pesando 54,85 ± 20,31kg. Foi instalado na veia jugular esquerda um introdutor e, posteriormente, posicionado um cateter de Swan-Ganz com a extremidade distal alocada no lumen da artéria pulmonar. Foi administrado pela via intramuscular detomidina (10μg/kg) associado a nalbufina (0,1mg/kg). Foram avaliadas FC, PAS, PAD, PAM, PVC, PAPm, IC, IS, IRVS, FR, pH, PaO2, PaCO2, HCO3, TC, sedação e motilidade ruminal antes do início da administração dos fármacos (MB) e a cada quinze minutos após a aplicação durante sessenta minutos (M15, M30, M45 e M60). Houve redução do IC, FR e aumento da PAS, PAPm, temperatura central, PaCO2 e HCO3 após administração dos fármacos. A sedação foi considerada satisfatória durante 45 minutos. Com os resultados obtidos neste estudo, conclui-se que a neuroleptoanalgesia promovida pela associação de detomidina e nalbufina em ovinos, nas doses utilizadas, promove sedação satisfatória. As alterações hemodinâmicas, respiratórias e na motilidade ruminal observadas podem ser bem toleradas por animais sadios. / Abstract: The aim of this study was to evaluate the effects on hemodynamics, respiration, ruminal motility and sedation of the combination of detomidine and nalbuphine in sheep. Were used eight healthy young, female or male sheep, weighing 54.85 ± 20.31kg. A Percutaneous Sheath Introducer was placed in the left jugular vein and then a Swan-Ganz catheter was positioned with the distal port allocated to the lumen of the pulmonary artery. Association of detomidine (10µg/kg) and nalbuphine (0,1mg/kg) was administered intramuscular. HR, SAP, DAP, MAP, CVP, MPAP, CI, SI, SVRI, RR, pH, PaO2, PaCO2, HCO3, CT, sedation and ruminal motility before drug administration (MB) and at each fifteen minutes after application for sixty minutes (M15, M30, M45 and M60). There was a reduction in CI, RR and increase in SAP, mPAP, CT, PaCO2 and HCO3 after drug administration. Sedation was considered satisfactory for 45 minutes. The results of this study allowed us to conclude that neuroleptoanalgesia promoted by the association of detomidine and nalbuphine in sheep at the doses used, promotes satisfactory sedation for short procedures. The hemodynamic, respiratory and ruminal motility changes observed can be well tolerated by healthy animals. / Mestre

No description available.

alpha2-adrenerge Rezeptoren, von denen drei verschiedene Subtypen (alpha2A, alpha2B, alpha2C) kloniert wurden, gehören zur Familie der G-Protein-gekoppelten Rezeptoren und vermitteln vielfältige physiologische Funktionen der Transmitter Adrenalin und Noradrenalin. Im Rahmen dieser Arbeit sollte untersucht werden, inwieweit Rezeptorsubtypen, die subzelluläre Lokalisation von Rezeptoren oder der Differenzierungsstatus einer Zelle für die funktionelle Diversität der alpha2-Rezeptor-Effekte in vivo verantwortlich sind. Im ersten Teil des Projektes wurde ein transgenes Mausmodell untersucht, bei dem selektiv alpha2A-Rezeptoren unter Kontrolle des Dopamin-beta-Hydroxylase Promotors in adrenergen Neuronen exprimiert wurden. In diesem Modell sollte getestet werden, ob ein einzelner Rezeptorsubtyp in den verschiedenen Neuronen des sympathischen Nervensystems in vivo identische Funktionen hat. Transgene alpha2A-Rezeptoren hemmten in vivo zwar die Freisetzung von Noradrenalin aus sympathischen Nervenfasern nicht aber die Exozytose von Adrenalin aus dem Nebennierenmark. Deshalb stellte sich die Frage, ob die Rezeptorfunktion von der Morphologie, dem Differenzierungsstatus der Zellen oder von der subzellulären Lokalisation der Rezeptoren abhängt. Hierfür wurden alpha2A-Rezeptoren durch Varianten des grün fluoreszierenden Proteins markiert und mittels FRET-Fluoreszenzmikroskopie untersucht. In PC12 Phäochromozytomzellen, die durch NGF zum Auswachsen neuronaler Fortsätze stimuliert wurden, waren die Agonist-bedingten Konformationsänderungen von alpha2A-Rezeptoren jedoch weder vom Differenzierungsstatus der Zellen noch von deren subzellulärer Lokalisation abhängig. Lediglich in transient transfizierten Zellen waren im Vergleich zu stabil transfizierten Zellen höhere Agonist-Konzentrationen zur Rezeptoraktivierung erforderlich. Diese Befunde zeigen, dass zusätzlich zur Diversität der Rezeptorsubtypen auf Proteinebene der zelluläre Kontext, in dem ein Rezeptor exprimiert wird, eine ganz wesentliche Rolle für dessen Funktion spielt. / alpha2-adrenergic receptors of which three different subtypes were cloned (alpha2A, alpha2B, alpha2C), are part of the family of G-protein coupled receptors and mediate many physiological functions of the transmitters adrenaline and noradrenaline. This study was initiated to determine whether receptor subtypes, their subcellular localization or the status of differentiation of a cell are responsible for the functional diversity of effects of alpha2-adrenergic receptors in vivo. In the first part of this project a transgenic mouse model was characterized, in which alpha2-adrenergic receptors were expressed under control of the dopamine-beta-hydroxylase-promotor in adrenergic neurons selectively. This model was used to test whether a single receptor subtype has identical functions in different neurons of the sympathetic nervous system in vivo. Transgenic alpha2A-adrenergic receptors inhibited the release of noradrenaline from sympathetic nerves in vivo, but not the exocytosis of adrenaline from the adrenal medulla. Therefore the question arose whether the functions of receptors are dependent on cell morphology, the status of differentiation of cells or the subcellular localization of receptors. To address this question, alpha2A-receptors were tagged with variants of the green fluorescent protein and investigated by means of FRET fluorescence microscopy. In PC12 rat pheochromocytoma cells which were stimulated by nerve growth factor to develop neurites, the conformational changes of alpha2A-adrenergic receptors upon agonist activation, however, did not dependent on the status of cellular differentiation or on the subcellular localization of receptors. Only in transiently transfected cells, higher agonist concentrations were necessary for the activation of receptors as determined by FRET microscopy. These findings demonstrate that the cellular context in which receptor subtypes are expressed play an essential role for their function.

Retinal ganglion cells play a crucial role in the relay of visual signals from the eye to the brain. This cell type is affected and eventually lost in the eye disease glaucoma, resulting in progressive and irreversible loss of vision. Studies of the molecular mechanisms leading to retinal ganglion cell death are important for the understanding of the disease and for designing future treatments. This thesis addresses and studies these molecular mechanisms, including alterations in gene expression after experimental retinal injuries. The effects of a neuroprotective drug, brimonidine, after transient retinal ischemia were also studied in order to help explain the mechanisms behind the protective properties of this drug. Several methods, including quantitative reverse transcriptase PCR, micro-arrays, western blot and immunohistochemistry, were used. The results showed that transient retinal ischemia triggers cell division in Müller cells and alters the gene expression of growth factors, their receptors, and intermediate filaments in the retina. Several genes related to the apoptosis process were less affected. Pre-treatment with brimonidine increased the levels of certain growth factors (BDNF, NT3, CNTF, FGF9) compared with vehicle. Brimonidine also had marked effects on genes related to progenitor cells, among them the recognized neural stem cell marker nestin. The increase in levels of nestin after ischemia was countered by brimonidine treatment. Moreover, retinal ganglion cell death following either optic nerve transection or optic nerve crush appears to involve the extrinsic apoptotic pathway although the gene expression response appears to differ between these injuries. The results obtained in this work contribute to an increased understanding of retinal injuries and highlight the importance of Müller cells in the endogenous defense against retinal injuries.

Thesis (Ph.D. in Clinical Science) -- University of Colorado Denver, 2007. / Typescript. Includes bibliographical references (leaves 130-142). Free to UCD affiliates. Online version available via ProQuest Digital Dissertations;

Diabetic kidney disease (DKD) is the leading cause of end stage renal disease worldwide and occurs in up to 40% of patients with diabetes. The standard of care for DKD treatment has not kept up with the current health epidemic, which has led to a heavy economic toll and substantial health burden. Targeting either cell surface (cs)GRP78, activated α2-macroglobulin (α2M*) or preventing their interaction may provide a novel anti-fibrotic therapeutic target for the treatment of DKD and potentially non-diabetic chronic kidney disease (CKD) as well. Previously our lab has shown that HG-induced csGRP78 is a mediator of PI3k/Akt signaling and downstream extracellular matrix (ECM) protein production in glomerular mesangial cells (MC). However, the ligand responsible for activating high glucose (HG)-induced csGRP78 had not yet been determined. We have shown thus far that α2M is endogenously produced, secreted, and activated (denoted α2M*) in HG by MC, which leads to its binding to and activation thereof csGRP78. Further, α2M knockdown or α2M* neutralization attenuated Akt activation, the production of the profibrotic cytokine connective growth tissue factor (CTGF) and ECM proteins fibronectin and collagen IV. We have also shown that integrin β1 (Intβ1), a transmembrane receptor, associated with csGRP78 under HG conditions and likely acts as a tether to present csGRP78 completely extracellularly on MC. Interestingly, Intβ1 activation, even in the absence of HG, was sufficient to induce csGRP78 translocation. Further, inhibition of either csGRP78 or Intβ1 prevented synthesis, secretion and signaling of TGFβ1. This data implicates a role for Intβ1 as a required signaling partner for csGRP78-mediated profibrotic signaling. To further our understanding of csGRP78/ α2M*’s role in DKD, we investigated their ability to mediate TGFβ1 signaling through its non-proteolytic activator thrombospondin-1 (TSP1). Here, HG-induced TSP1 expression, ECM deposition, and activation of TGFβ1 was regulated by the PI3k/Akt pathway via csGRP78/α2M* in MC. Furthermore, we assessed whether this csGRP78/ α2M* axis is relevant to promoting profibrotic signaling in other renal cell types, including proximal tubule epithelial cells (PTEC) and renal fibroblasts (RF), that contribute to the pathogenesis of both later stage DKD and non-diabetic CKD. We show evidence here that HG and direct treatment with TGFβ1, a key pathologic regulator of kidney fibrosis, induce GRP78 surface translocation as well as the endogenous production and activation of α2M in both PTEC and RF. Inhibition of either csGRP78 or α2M* prevented TGFβ1 signaling measured as Smad3 activation as well as downstream ECM production. Interestingly, inhibition of this pathway under direct TGFβ1 treatment did not prevent Smad3 activation, implicating a role for Smad-independent TGFβ1 signaling through this axis. We identified the known noncanonical TGFβ1 signaling partners, yes associated protein (YAP) and transcriptional co-activator with PDZ binding motif (TAZ), are mediated by csGRP78 and α2M*. Lastly, we evaluated the potential therapeutic benefit of inhibiting csGRP78/α2M* interaction in the kidney fibrosis model, unilateral ureteral obstruction (UUO). Here, we show evidence that inhibition of this signaling axis using an inhibitory peptide can prevent renal fibrosis. Whether this peptide also prevents fibrosis in DKD is currently being assessed. Together, these studies strongly implicate targeting csGRP78/α2M* interaction as a novel anti-fibrotic therapeutic intervention for early and late stage DKD, as well as a potential role in non-diabetic CKD. / Thesis / Doctor of Philosophy (Medical Science) / Diabetic kidney disease is the leading cause of kidney failure in developed nations. This progressive disease leads to the loss of kidney function due to an accumulation of scar proteins in the kidney over time. High glucose is a major factor that causes this to occur. Our lab studies specific kidney cells called mesangial cells, proximal tubule epithelial cells, and fibroblasts that produce scar proteins in the presence of high glucose. We have shown that when these cells are treated with high glucose, this causes the movement of a protein called GRP78 that normally resides inside the cell to move to the cell’s surface where it can interact with other proteins. My research has established that the proteins alpha 2-macroglobulin (ɑ2M), integrin β1 (Intβ1), and thrombospondin-1 (TSP1) can bind to GRP78 on the cell surface and cause cells to make scar proteins. Preventing ɑ2M or Intβ1 from binding to GRP78 or preventing TSP1 production prevents mesangial cells from making scar proteins when exposed to high glucose. In a mouse model that overproduces these scar proteins, we showed that preventing cell surface GRP78 and α2M interaction prevents scar protein production and is thus a novel potential treatment option for kidney disease.

<p>Grade 2 gliomas are malignant brain tumours affecting otherwise healthy adults. Although the long-term prognosis is poor, many patients are well and may have a high quality of life for several years. There is, however, a large variability in the natural course of the disease which makes it essential to identify patients who might benefit from early surgery or radio-therapy. The aim of the present thesis was to define new and clinically useful prognostic markers that may assist in the initial treatment decision and in patient follow-up.</p><p>A retrospective study of 189 patients with gliomas WHO grade 2 showed no advantage in survival of early tumour resection or radiotherapy, and confirmed that histological subtype and patient age are the most important predictors of survival (I). In 89 patients, the pre-treatment uptake of 11C-methionine (MET) measured with positron emission tomography (PET) was identified as a prognostic marker for survival (II). At the time of tumour progression, irradiated tumours demonstrated signs of a residual radiotherapeutic effect that correlated with the pre-treatment uptake of MET (III). Pre-treatment uptake of MET may, therefore, be important both in predicting the natural course of the disease and the response after treatment. Immunohistochemical staining of 40 tumour samples showed an inverse association between the number of tumour cells expressing platelet-derived growth factor alpha receptor (PDGFRa) and survival (IV). Also, a reduction was observed in the number of receptor-positive cells after malignant transformation, supporting the prognostic value of PDGFRa.</p><p>Lumbar puncture was performed in eight patients with newly diagnosed low-grade gliomas to identify three important growth factors in tumour development. Neither PDGF nor vascular endothelial growth factor (VEGF) were detected in the cerebrospinal fluid (CSF), and fibroblast growth factor 2 (FGF-2) was measurable at extremely low concentrations in two of the patients (V). A proteome screening of the CSF, using two-dimensional gel electrophoresis and mass spectrometry, detected alpha 2-HS glycoprotein at significantly higher concentrations than in a control group (VI). This glycoprotein emerges as a novel substance in glioma research and may be of great interest because of its suggested involvement in the embryonic development of the neocortex.</p>

Grade 2 gliomas are malignant brain tumours affecting otherwise healthy adults. Although the long-term prognosis is poor, many patients are well and may have a high quality of life for several years. There is, however, a large variability in the natural course of the disease which makes it essential to identify patients who might benefit from early surgery or radio-therapy. The aim of the present thesis was to define new and clinically useful prognostic markers that may assist in the initial treatment decision and in patient follow-up. A retrospective study of 189 patients with gliomas WHO grade 2 showed no advantage in survival of early tumour resection or radiotherapy, and confirmed that histological subtype and patient age are the most important predictors of survival (I). In 89 patients, the pre-treatment uptake of 11C-methionine (MET) measured with positron emission tomography (PET) was identified as a prognostic marker for survival (II). At the time of tumour progression, irradiated tumours demonstrated signs of a residual radiotherapeutic effect that correlated with the pre-treatment uptake of MET (III). Pre-treatment uptake of MET may, therefore, be important both in predicting the natural course of the disease and the response after treatment. Immunohistochemical staining of 40 tumour samples showed an inverse association between the number of tumour cells expressing platelet-derived growth factor alpha receptor (PDGFRa) and survival (IV). Also, a reduction was observed in the number of receptor-positive cells after malignant transformation, supporting the prognostic value of PDGFRa. Lumbar puncture was performed in eight patients with newly diagnosed low-grade gliomas to identify three important growth factors in tumour development. Neither PDGF nor vascular endothelial growth factor (VEGF) were detected in the cerebrospinal fluid (CSF), and fibroblast growth factor 2 (FGF-2) was measurable at extremely low concentrations in two of the patients (V). A proteome screening of the CSF, using two-dimensional gel electrophoresis and mass spectrometry, detected alpha 2-HS glycoprotein at significantly higher concentrations than in a control group (VI). This glycoprotein emerges as a novel substance in glioma research and may be of great interest because of its suggested involvement in the embryonic development of the neocortex.