Orientadores: Maria Alice da Cruz-Hofling, Stephen Hyslo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-03T17:09:30Z (GMT). No. of bitstreams: 1 Silva_GustavoHenriqueda_M.pdf: 4972230 bytes, checksum: 422b67c158699f0d6902a1376fbb2548 (MD5) Previous issue date: 2003 / Resumo: Envenenamentos humanos por lagartas da mariposa Lonomia obliqua (Saturniidae) que ocorrem no sul do Brasil produzem uma moderada resposta local (eritem~ edema e dor) e efeitos sistêmicos que incluem incoagulabilidade sangüíneo insuficiência renal e sérios acidentes hemorrágicos intracerebrais. Através de estudos quantitativos, histológicos, imunohistoquímicos e ultraestruturais foram analisadas as alterações morfológicas e a quebra da barreira hematoencefálica (BlIE) em ratos injetados intravenosamente (Lv.) com o veneno do extrato das espículas de L. obliqua (200 mglkg). Além disso, foi avaliada a biodistribuição do veneno no figado, rins e cérebro. Em relação à quebra de BlIE foram estudadas também cinco frações semipurificadas (Sephadex 075) do veneno (200 m glkg cada). Análises quantitativas morfológicas, imunohistoquímicas e ultraestruturais foram feitas 6, 18, 24 e 72 h após a injeção (p.L) Lv. do veneno e das fIações. A microscopia de luz mostrou que 6 h após o envenenamento havia edema cerebral, que diminuía em 72 h. Hemorragia intracerebral ocorreu apenas em um rato 24 horas após a injeção do veneno. A imunomarcação da proteína glial fibrilar ácida (OF AP) mostrou um aumento progressivo em sua expressão nos grupos de 6, 18, 24 e principalmente 72 h. A biodistribuição do veneno 6 e 18 h após injeção Lv. evidenciado pela imunohistoquímica mostrou que apenas no figado (6 h p.L) e nos rins (6 e 18 h p.i.) foram detectadas quantidades de veneno, o que não foi observado no cérebro. A quebra da BlIE, observada por microscopia eletrônica de transmissão, evidenciada pela passagem do traçador extracelular (nitrato de lantânio) através das células endoteliais do capilar cerebral. A ruptura da BlIE foi observado no cerebelo e no hipocampo 18 h após a injeção do veneno. Neste período, o cerebelo foi mais sensível ao veneno que o hipocampo, exibindo um número maior de vasos afetados. O número de capilares mostrando quebra foi menor após 72 h do que quando comparada ao grupo de 18 h. Nenhuma das fIações semi-purificadas aumentou significativamente a permeabilidade dos vasos. Estes resultados indicam que o. veneno de L. obliqua apresenta uma ação deletéria sobre a BlIE de ratos. O aumento na expressão da OF AP indica reatividade astrocitária provocada pela ação do veneno. A ausência de efeitos das frações semi-purificadas quando administradas separadamente, sugerem uma ação sinérgica dos componentes do veneno, podendo ser responsável pelos danos observados no Sistema Nervoso Central (SNC). A cinética da distribuição do veneno nos órgãos estudados está de acordo com o metabolismo de xenobióticos exercido pelo fígado e com a eliminação destes promovida pelos rins. A não detecção do veneno no cérebro, apesar da quebra da BHE, sugere que o veneno pode não entrar neste órgão nos períodos observados. Os distúrbios morfológicos permaneceram após a eliminação de veneno, explicando a longa duração dos sintomas do lonomismo / Abstract: Human envenoming by caterpillars of the saturniid moth Lonomia obliqua in southem Brazil produces a mild local response (erythema, some oedema, and pain) and systemic effects which include incoagulable blo~ renal failure and in severe accidents intracerebral hemorrhage. In this work, we used light and electron microscopy to investigate the morphological alterations in the brain and blood-brain barrier of rats injected intravenously with venom from L. obliqua spicules (200 mg/kg). Five semi-purified fractions ofvenom (200 mg/kg each) were also assayed. Quantitative morphological, ultrastructural and imunohistochemical analyses were done 6 to 72 h after the L v. injection of venom and its fractions. Light microscopy showed that 6 h after envenoming there was cerebral edema, which decreased by 72 h. Intracerebral hemorrhage occurred in only one rat 24 h after the injection of venom. The imunohistochemistry of the Glial Fibrillary Acidic Protein (GFAP) showed a progressive increase in its expression at 6, 18,24 and mainly 72 h. The venom biodistribution 6 and 18 h after Lv. injection (observed by imunohistochemistry) showed venom immunolabeling in the liver, 6 h post injection (p.L), and kidneys (6 and 18 h p.i.), however in the brain it was not possible to observe probably because the blood-brain barrier (BBB) specificity. BBB breakdown, assessed by transmission electron microscopy based on the passage of an extracellular tracer (lanthanum nitrate) between brain capillary endothelial cells, was observed in the cerebellum and hippocampus 18 h after venom injection. At this time, the cerebellum was more sensitive to the venom than the hippocampus, as shown by the greater number of Ieaky vessels. The number of capillaries showing breakdown was lower after 72 h than after 18 h. None of the semi-purified fractions significantly increased the number of Ieaky vesseIs. These results indicate that L. obliqua caterpillar venom has a deIeterious action on the rat BBB. The expression increase of GF AP astrocyte reactivity therefore that venom has a neurotoxic action. The Iack of effect of the venom fractions when administered alone suggests that a synergistic action of the venom components may be responsibIe for the damaf;!e in the Central Nervous System (CNS) after the whole venom injection. After 6 h for the liver and 18 h for the kidneys alI the venom brought by the systemic circulation seems to be entirely vanished from these organs, but morphological disturbances remained what explain the long lasting symptoms of the lonomism / Mestrado / Mestre em Farmacologia

Orientador: Edson Antunes / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-07-25T19:01:13Z (GMT). No. of bitstreams: 1 Penteado_CarlaFernandaFranco_M.pdf: 2582288 bytes, checksum: b84812ec12fc6c20a7e59cc8b9799a0b (MD5) Previous issue date: 1999 / Resumo: Neste trabalho investigamos a participação do NO no edema de pata, permeabilidade vascular e influxo de neutrófilos para cavidade peritoneal induzidos pela enterotoxina estafilocócica tipo B (SEB) em camundongos. Para tanto, utilizamos inibidores não-seletivos (L- NAME) e seletivos (aminoguanidina e 7 -nitroindazol) da NO-sintase, assun como a dexametasona (inibidor da indução da iNOS). Nossos resultados mostraram que o L-NAME (7-28 mg!kg, i.v.) e aminoguanidina (25-75 mg!kg, i.v.) reduziram significativamente o edema de pata induzido pela SEB (25 ~g/pata). A administração subplantar de tais inibidores também causou significante redução do edema de pata. Por outro lado, apenas as maiores doses destes inibidores (administrados pela via endovenosa) foram capazes de inibir de maneira significativa, o extrayasamento de proteínas induzido pela toxina. O D- NAME (enântiomero inativo), administrado local ou sistemicamente, não alterou a resposta edematogênica induzida pela SEB. A inibição do edema de pata causada pelo L-NAME e aminoguanidina foi revertida pelo iloprost (análogo da prostaciclina), sugerindo que o efeito inibitório destes inibidores deve-se, provavelmente, à redução de fluxo sangiiíneo local como conseqüência da inibição do NO. Aadministração sistêmica do L-NAME (14-56 mg!kg), aminoguanidina (25-75 mg!kg) ou dexametasona (0,25-1,0 mg!kg) atenuou o influxo de neutrófilos induzido pela SEB em cavidades peritoneais com número de macrófagos normais. Em cavidades com o número de macrófagos aumentado, os inibidores foram menos eficazes em reduzir o influxo dessas células. O D-NAME não afetou o influxo de neutrófilos induzido pela SEB nas duas cavidades estudadas. Os resultados da medida da atividade da NOS mostraram que a nNOS do cérebro foi significativamente inibida pelo L-NAME 4 (28 mg/kg) e 12 h (14-56 mg/kg) após administração endovenosa deste inibidor. A aminoguanidina (75 mg/kg, i.v.) reduziu a atividade da nNOS em 4 h, mas não em 12 h após a administração sistêmica. O 7-nitroindazol (5-15 mg/kg) não modificou a nNOS em quaisquer das doses utilizadas. A atividade da iNOS foi inibida significativamente pelo L-NAME (14-56 mg/kg, i.v., 12 h) e aminoguanidina (25-75 mg/kg, i.v., 12 h). Concluímos que o NO é um mediador importante na resposta edematogênica e no influxo de neutrófilos induzidos pela SEB. A síntese do NO, no sítio inflamatório, parece envolver a cNOS na fase inicial (4 h) e a iNOS na fase de mais tardia (12 h) / Abstract: In this study we attempted to investigate the role of nitric oxide (NO) on the inflammatory responses (hindpaw oedema, vascular permeability and neutrophil migration) induced by the staphylococcal enterotoxin type B (SEB) in mouse. To achieve this, both non-selective (L-NAME) and selective (aminoguanidine and 7-nitroindazole) NO-synthase (NOS) inhibitors as well as dexamethasone (iNOS induction inhibitor) were used. Our results showed that L-NAME (7-28 mg/kg, i.v.) and aminoguanidine (25-75 mg/kg, i.v.) significantly reduced SEB-induced paw oedema. Similar results were observed when these inhibitors were administered intraplantarly. The inactive enantiomer D-NAME had no effect on the SEB-induced oedema formation. The inhibition of paw oedema by both L- NAME and aminoguanidine were significantly reversed by the local . administration of the prostacyclin analogue iloprost. This suggest that these inhibitors probably acts by reducting regional blood flow. The compound 7¬nitroindazole, a substance which preferentially inhibit nNOS, had no effect on the exsudation and paw oedema induced by SEB. L-NAME and aminoguanidine as well as dexamethasone significant reduced neutrophil accumulation induced. by SEB into mouse peritonenl cavity. In thioglycollate-pretreated cavities, a condition where the number of macrophages is increased, L-NAME and aminoguanidine caused a smaller inhibition of SEB-induced neutrophil migration, as compared to naive cavities. D-NAME had no effect on the neutrophil migration induced by SEB, either in non-stimulated or thioglycollate-treated cavities. In addition, bNOS activity was reduced by L-NAME at 4 h (28 mg/kg) and 12 h (14-56 mg/kg) after endovenous administration, while aminoguanidine (75 mg/kg, i. v.) was able to reduce the nNOs activity only when this inhibitor was injected 4 h before. The nNOS activity was unchanged by 7 -nitroindazole. The activity of iNOS was significant1y inhibited by L-NAME (14-56 mg/kg, i.v., 12 h) and aminoguanidine (25-75 mg/kg, i.v., 12 h).In conclusion, our results indicate that NO modulate both oedema and neutrophil migration induced by SEB. We suggest that NO involved in inflammatory response was produced by cNOS in the early phase (4 h) and iNOS during the sustained phase (12 h) / Mestrado / Farmacologia / Mestre em Ciências Médicas

Ankle taping and bracing are important components in the world of sports medicine and athletic training. There are different types of tape that are used for athletes by athletic trainers. However, little research has compared the performance of Kinesio™ Tape and an ASO ankle brace on a lateral grade I ankle sprain. The purpose of this study was to determine if there are therapeutic effects of Kinesio™ tape on a grade I lateral ankle sprain, using an ASO as the control group. The gold standard protocol for a grade I lateral ankle sprain is an ASO ankle brace.Twenty five students from 9 different high schools in 2 sister counties participated in the study. These students were all athletes, who suffered a grade I lateral ankle sprain. Twelve student-athletes were in the Kinesio™ tape group and thirteen of the student-athletes were in the ASO brace group, also known as the control group. The student-athletes completed five functional tests; Single Leg Stance, Single Leg Squat, Single leg Hop, Box Drill , and the Illinois Test, at each of the three sessions 0, 4, and 8 weeks.Results found there was no significant difference between the Kinesio™ tape group and the ASO™ ankle brace group when it came to pain level, single leg hop for distance, the box drill or the Illinois test, when tested at three different times; week 0, week 4, and week 8. However, results showed that there was a significant difference between the two groups on the single leg squat test. It was determined that the control group also known as the ASO™ ankle brace group, were able to perform more squats at week 4 and week 8, than the treatment groups also known as the Kinesio™ Tape group. / Ph. D.

No description available.

Introduction Retinal vascular reactivity is impaired in patients with diabetes and is thought to be involved in the onset and progression of diabetic retinopathy (DR). Previous studies that have utilized hyperoxia to assess retinal vascular reactivity have been limited due to confounding factors associated with the administration of oxygen and have used a variety of different instruments to measure retinal blood flow. The influence of blood glucose at the time of blood flow assessment has also not been systemically investigated. The specific aims of each Chapter are as follows: Chapter 3: To compare three systems used to administer hyperoxia to human subjects. Chapter 4: To quantify the magnitude and timeline of change of retinal hemodynamic parameters induced by an isocapnic hyperoxic stimulus. Chapters 5, 6 and 7: To quantify the magnitude of change of retinal hemodynamic parameters induced by hyperoxia, hyperglycemia and combined hyperoxia / hyperglycemia, respectively, in groups of diabetic patients with no clinically visible, and mild-to-moderate, DR and in age-matched subjects without diabetes. Methods Chapter 3: Subjects breathed air followed by oxygen, or oxygen plus carbon dioxide using a non-rebreathing system, or air followed by oxygen using a sequential rebreathing system. The magnitude of change and variability of CO2 concentrations was compared between systems. Chapter 4: Baseline retinal blood flow data was acquired while the subjects breathed air using a sequential rebreathing system. An isocapnic hyperoxic stimulus was initiated and maintained for 20 minutes. Air was then re-administered for 10 minutes. Retinal blood flow measurements were acquired every minute over the course of the study. The magnitude of change of each hemodynamic parameter was determined by fitting individual data with a sigmoidal function. For Chapter 5, 6 and 7 diabetic patients with no clinically visible, and mild-to-moderate, DR were stratified into groups based upon their retinopathy status. Age-matched non-diabetic subjects were recruited as controls. Baseline retinal blood flow data was acquired while subjects breathed air. Retinal blood flow measurements were then acquired after exposure to (a) hyperoxia, (b) hyperglycemia and (c) combined hyperoxic / hyperglycemic stimuli. Change in hemodynamic parameters was compared between groups and correlated with objective measures of retinal edema. Results Chapter 3: The difference in group mean end-tidal CO2 levels between baseline and hyperoxia was significant for oxygen administration using a non-rebreathing system. The sequential rebreathing technique resulted in a significantly lower variability of individual CO2 levels than either of the other techniques. Chapter 4: An ~11% decrease of diameter, ~36% decrease of velocity and ~48% decrease of blood flow was observed in response to isocapnic hyperoxia in young, healthy subjects. A response time of 2.30??0.53 minutes and 2.62??0.54 minutes was observed for diameter and velocity, respectively. Chapter 5: Retinal blood velocity, flow, and WSR significantly decreased in response to isocapnic hyperoxia in all groups. The magnitude of the reduction of blood flow was significantly reduced with increasing severity of retinopathy. There was a significant relationship between baseline objective edema index values and retinal vascular reactivity. Chapter 6: A significant change in blood glucose level was observed for all groups. No significant change in any hemodynamic parameter was found in patients with diabetes and in age-matched subjects without diabetes. Chapter 7: Retinal blood velocity and flow significantly decreased in all groups in response to combined hyperoxic / hyperglycemic provocation. The vascular reactivity response was not significantly different across the groups. Conclusions Chapter 3: Control of CO2 is necessary to attain standardized, reproducible hyperoxic stimuli for the assessment of retinal vascular reactivity. Chapter 4: Arteriolar retinal vascular reactivity to isocapnic hyperoxic provocation occurs within a maximum of 4 minutes. Although there was a trend for diameter to respond before velocity, the response characteristics were not significantly different between diameter and velocity. Different response characteristics of the retinal vasculature to transmural pressure mediated autoregulation as opposed to metabolic mediated vascular reactivity are suggested. Chapter 5: The vascular reactivity response in terms of the reduction of blood flow relative to baseline was significant in all groups but the magnitude of the change in flow was significantly reduced with increasing severity of retinopathy. A loss of retinal vascular reactivity is indicated in patients with moderate DR without clinically evident diabetic macular edema (DME), and in patients with DME. Chapter 6: Unaltered retinal arteriolar blood flow was found 1 hour after glucose ingestion in patients with diabetes and in age-matched subjects without diabetes. These results do not support the theory that retinal blood flow is affected by an acute increase of blood glucose in diabetic patients and in subjects without diabetes. Chapter 7: The vascular reactivity response to a combined hyperoxic / hyperglycemic provocation produced a pronounced reduction in blood flow. Unlike the response to hyperoxia alone, the vascular reactivity response was not significantly different across the groups. This suggests that hyperglycemia may influence the retinal vascular reactivity response to hyperoxia.

No description available.

PURPOSE: Diabetic macular edema (DME) is the most common cause of vision loss in patients with diabetic retinopathy. This study aims to approach diabetic macular edema and diabetic retinopathy as ischemic conditions and explores a potential treatment through hyperoxia. The study measured changes in retinal thickness, visual acuity, and contrast sensitivity in subjects receiving normobaric oxygen. METHODS: Fifty-one patients with diabetic macular edema at Beth Israel Deaconess Medical Center Eye Clinic (Boston, MA) received oxygen via a face mask at 5 liters per minute for 3 hours. Retinal thickness at the central subfield and maximal retinal thickness were measured using optical coherence tomography. Contrast sensitivity, best corrected visual acuity, and intraocular pressure were measured before and after oxygen as well. RESULTS: Macular thickness from diabetic macular edema decreased by an average of 2.09% (p < .05) at the point of maximal thickness, and by 0.88% (p < .05) at the central subfield. Vision also improved by an average of 0.043 LogMAR units (p < .05). Changes in macular thickness and visual acuity were non-significant in healthy control eyes that received oxygen. The results of hyperoxia on contrast sensitivity were indeterminate. CONCLUSIONS: We found that normobaric hyperoxia for 3 hours reduces macular thickness from diabetic macular edema and improves visual acuity. This study offers additional evidence that diabetic macular edema is an ischemic disorder and suggests that oxygen therapy may serve as an alternate or complimentary treatment of DME. / 2020-06-17T00:00:00Z

Introdução Pacientes com apneia obstrutiva do sono exibem alterações morfológicas encefálicas. A hipóxia intermitente causa morte celular em 90% dos neurônios de Purkinje. Aquaporinas são proteínas de membrana que formam canais de água. O conhecimento do papel das aquaporinas nas alterações encefálicas causadas pela apneia obstrutiva do sono é limitado. Objetivos Avaliar o efeito da hipóxia intermitente isocápnica nas aquaporinas e no dano celular em diferentes tecidos de camundongos C57/bl. Métodos Conteúdo cerebral de água e expressão de aquaporinas 1, 3, 4, 9, e HIF-1a foram medidos. Realizou-se coloração com hematoxilina-eosina e marcação por imunohistoquímica, além de técnicas de Western blot e ELISA. Resultados Os níveis de HIF-1a no córtex frontal foram maiores no grupo exposto a hipóxia intermitente isocápnica do que no grupo controle, confirmando hipóxia efetiva. O peso molhado e o conteúdo de água do encéfalo foram maiores no grupo hipóxia. Os níveis cerebelares de aquaporina 1 foram menores no grupo hipóxia. Os níveis de aquaporina 3 no hipocampo e pele foram similares em ambos os grupos. Aquaporina 4 no córtex frontal foi mais expressa no grupo hipóxia. Aquaporina 9 apresentou maiores níveis no córtex frontal e estriado do grupo hipóxia. Observou-se dano celular em 88% das células de Purkinje no grupo hipóxia. A marcação de S100B no cerebelo de grupo hipóxia foi mais evidente do que nos controles. Conclusão Exposição crônica a hipóxia simulando apneia do sono causa edema encefálico e altera a concentração de aquaporinas. Alterações em proteínas de canal de água podem estar envolvidas no mecanismo do dano observado em estruturas cerebrais na apneia obstrutiva do sono. / Background Patients with obstructive sleep apnea exhibit cerebral morphological changes. The damaging effect of intermittent hypoxia was already demonstrated in a study where mice exposed to this condition had 90% of degeneration of Purkinje cells. Aquaporins are membrane proteins responsible for the transport and the balance of water content in the brain. Little attention has been dedicated to the role of aquaporins in sleep apnea-related brain alterations. Objectives To evaluate the effect of isocapnic intermittent hypoxia on water-transport proteins and cellular damage in different tissues of C57/bl mice. Methods Brain water content, aquaporins 1, 3, 4, 9, and HIF-1a were measured. Staining with hematoxylin-eosin and immunohistochemistry were performed. Results HIF-1a expression in the frontal cortex was higher in the exposed than in the sham group, confirming effective hypoxia. The brain wet weight and brain water content were higher in the hypoxia group. Cerebellar Aquaporin-1 levels were lower in the hypoxia group. Aquaporin 3 levels in hippocampus and skin were similar in both groups. Aquaporin 4 expression in the frontal cortex was higher in the hypoxia group. Aquaporin 9 expression of the hypoxia group was greater in the cortex and striatum. Death rate of Purkinje cells in the hypoxia group was 88%. Stronger immunoreactivity to S100B was detected in the hypoxia group. Conclusions Chronic exposure to hypoxia simulating sleep apnea causes brain edema and alters the concentration of aquaporins. Water channel proteins are possibly implicated in the cellular damage to brain structures seen in obstructive sleep apnea.

The retina is one of the most metabolically active tissue in the body, rendering it sensitive to vascular dysfunction. Consequently, diseases that disrupt normal retinal blood supply, including retinal vein occlusions (RVO) and diabetic retinopathy, are the leading causes of blindness in working-age adults. Despite available therapies, an estimated 50% of patients do not respond to treatment. We employed a mouse model of retinal vein occlusion (RVO), achieved by tail-vein injection of rose bengal, followed by laser photocoagulation of retinal veins. In vivo analyses – optical coherence tomography (OCT), fluorescein angiography, and electroretinograms (ERGs) - were conducted with the Micron IV system (Phoenix Research Labs). RVO induces acute retinal edema, which peaks during the first 24 hours following injury. Over a 7 day time course the edema resolves, revealing a permanent retinal thinning due to death of retinal neurons. We identified caspase-9, a protease traditionally associated with apoptosis, as an essential mediator of edema. Increased levels of activated caspase-9 were detected in vascular endothelial cells 1 hour following RVO. We tested RVO in mice with inducible endothelial-cell-specific deletion of caspase-9 (iC9 ECKO). Compared to littermate controls, iC9 ECKO mice develop less edema, and sustain less retinal degeneration after RVO injury. ERG analysis showed preservation of retinal function in iC9 ECKO mice. To study whether inhibiting caspase-9 would provide protection against RVO we utilized a highly specific caspase-9 inhibitor, which we can deliver to the retina using simple eyedrops. Treatment of wildtype mice with the caspase-9 inhibitor immediately after induction of RVO provided morphologic, biochemical and functional retinal protection. Inhibition of caspase-9 reduces edema, protects retinal morphology, and helps prevent vision loss following RVO injury. Our studies indicate that endothelial caspase-9 plays an essential role in regulating edema pathogenesis. Moreover, our novel cell permeant caspase-9 inhibitor abrogates the edema and may be a potential therapy for individuals suffering from vascular eye disease.

The concept of genetic penetrance, "the frequency of manifestation of a genetic factor," was introduced by Timofeef-Ressovsky (Naturwissenschaften 19:493,1931). Incomplete penetrance has been used to explain the absence of phenotypic expression when otherwise anticipated. Studies of Embryonic Chick Edema, ECE (Poultry Sci. 77(suppl. 1):69, 1998) have been conducted in order to determine the origins for incomplete penetrance of this disorder. ECE was originally reported as the expression of two autosomal recessive loci with incomplete penetrance. Pedigreed inter se mating of ECE individuals have resulted in familial incidences ranging for 0 to 100% with a mean of 48.2% in the most recent generation selected. With consideration of a third contributing locus, current data and 2 sets of previous data were evaluated. Heterogeneity and pooled chi square tests when applied to the data sets support the hypothesis that ECE was the result of two completely dominant loci and one homozygous recessive locus. / Graduation date: 2004