Eclampsia is a hypertensive disorder of pregnancy and a leading cause of maternal death. The primary explanation for eclampsia is that it represents a form of hypertensive encephalopathy (HTE) with neurological symptoms including headaches, nausea, vomiting, visual disturbances, and seizures. The etiology of HTE involves an acute increase in arterial blood pressure that exceeds the autoregulatory capacity of the brain leading to forced dilatation of cerebral vessels, decreased cerebrovascular resistance, hyperperfusion, blood-brain barrier (BBB) disruption, and vasogenic cerebral edema formation. Due to the central role of the cerebral circulation in mediating these symptoms, a better understanding of how pregnancy affects the cerebral circulation is important to the treatment and prevention of eclampsia. A central goal of this dissertation was to determine pregnancy’s effect on cerebral blood flow (CBF) autoregulation, edema formation, and BBB permeability during acute hypertension. Women with eclampsia often seize at lower blood pressures than HTE patients. We hypothesized that pregnancy may predispose the brain to eclampsia by lowering the pressure of autoregulatory breakthrough and enhancing cerebral edema formation. Using an in vivo model of HTE, we found that the pressure of autoregulatory breakthrough was not different between nonpregnant (NP) and late-pregnant (LP) rats; however, cerebral edema formation was significantly increased only in LP animals. Nitric oxide synthase inhibition significantly increased the upper limit of autoregulation in both NP and LP animals and attenuated cerebral edema formation in LP animals. BBB permeability during acute hypertension was not different between these groups. Magnesium sulfate (MgSO4) is widely used to treat eclampsia despite an unclear mechanism of action. A second goal of this dissertation was to determine the cerebrovascular effects of MgSO4 during pregnancy. Specifically, we investigated the effect of MgSO4 on in vitro resistance artery vasodilation and in vivo BBB permeability during acute hypertension. We hypothesized that dilation to MgSO4 would be greater in mesenteric than cerebral vessels. MgSO4 elicited concentration-dependent vasodilation in all arteries, as determined by measuring lumen diameter of isolated and pressurized arteries, however, mesenteric arteries were considerably more sensitive than cerebral arteries. In addition, there was no effect of pregnancy on MgSO4 sensitivity in mesenteric arteries, whereas pregnancy decreased sensitivity to MgSO4 in cerebral arteries. We further hypothesized that MgSO4 would decrease BBB disruption during acute hypertension, thereby protecting the brain in eclampsia. Using an in vivo model of HTE, we showed that MgSO4 treatment decreased BBB permeability during acute hypertension in LP rats, with the greatest effect observed in the posterior cerebrum. In conclusion, this dissertation determined CBF autoregulation and cerebral edema formation during pregnancy, and also the effect of MgSO4 on cerebral resistance artery vasodilation and BBB permeability during acute hypertension in LP rats. Although pregnancy did not influence autoregulatory breakthrough, cerebral edema formation was enhanced in LP animals and this may potentiate neurological symptoms in eclampsia. In addition, MgSO4-induced cerebral vasodilation is likely not a primary mechanism of eclampsia treatment, rather MgSO4 may limit edema formation by attenuating BBB permeability during hypertension.

Memoria para optar al Título Profesional de Médico Veterinario / Las acciones estrogénicas de los fitoestrógenos han sido ampliamente estudiadas en las últimas décadas. Estos efectos agonistas y/o antagonistas estrogénicos en diversos órganos, despertó el interés de la comunidad científica en la medicina tradicional y ancestral. El presente estudio in vivo, se realizó en útero de ratas impúberes de 21 días de edad, tratadas con dos extractos preparados a partir de la misma planta medicinal, pero procesados de forma distinta. Las ratas fueron inyectadas subcutáneamente con cada uno de los extractos o su vehículo y/o 17β estradiol; los cuernos uterinos fueron extraídos 6 y 24 h post-tratamiento y luego procesados histológicamente. Las variables no genómicas evaluadas fueron: número de eosinófilos uterinos, su distribución dentro del útero, porcentaje de eosinófilos degranulados y edema en estroma endometrial profundo y superficial. Entre las respuestas genómicas se evaluó hipertrofia celular en miometrio circular. En ausencia de estradiol, los extractos 21a (solución hidroalcohólica) y 22a (solución etanólica) no inducen eosinofilia uterina, pero generan edema en estroma, aunque de menor magnitud que el generado por estrógeno. Los dos extractos generaron una hipertrofia de miometrio circular de magnitud comparable a la generada por estradiol. La administración del extracto 21a, previo a la administración de 17β estradiol, disminuyó drásticamente la eosinofilia, efecto que no fue generado por el extracto 22a. Sin embargo, con ambos extractos se evidenció una mayor proporción de eosinófilos en los tejidos más profundos de útero (miometrio circular y estroma) y una mayor proporción de degranulados, en comparación a lo observado con 17β estradiol. La administración de cada extracto asociado a 17β estradiol, generó edema endometrial de mayor magnitud al generado por estrógeno, para el caso del extracto 22a la máxima respuesta se observó a las 6 h y para el extracto 21a a las 24 h. Se observó un efecto potenciador de la hipertrofia celular de miometrio circular a las 24 h, luego de administrar los extractos asociados a 17β estradiol. El extracto 21a mostró la capacidad de generar respuestas no genómicas (edema) y genómicas (hipertrofia), e inhibir la eosinofilia en útero de rata prepúber, por lo que posee algún principio activo con capacidad de disociar respuestas estrogénicas. Se hace necesario evaluar otras respuestas estrogénicas consideradas de riesgo, tales como hiperplasia uterina para visualizar potenciales aplicaciones terapéuticas.

A natureza sempre despertou no homem um fascínio encantador, não só pelos recursos oferecidos para sua alimentação e manutenção, mas também sua principal fonte de inspiração e aprendizado. Allophylus racemosus Sw., objetivo deste trabalho, não apresenta relatos de sua utilização, e tampouco quando se trata de dados científicos em bases de pesquisa sobre estudos de suas atividades farmacológicas. Esta planta pertence à família Sapindaceae que compreende cerca de 140 gêneros e 1900 espécies pelo mundo todo. Dentre eles, o gênero Allophylus apresenta cerca de 250 espécies. Apesar de um número considerável de espécies, há uma escassa quantidade de pesquisas relacionadas a este gênero, e tampouco dessa espécie. As folhas dessa planta foram coletadas na região da mata atlântica mineira nas proximidades do município de Alfenas-MG. Para o estudo fitoquímico e teste de atividade anti-inflamatória, o extrato bruto etanólico das folhas de Allophylus racemosus Sw. foi obtido por maceração em etanol e submetida ao teste de edema de pata induzido por carragenina e edema de orelha, onde se constatou uma atividade anti-inflamatória estatisticamente significativa frente aos controles negativo e positivo (indometacina) utilizados. Em seguida realizou-se o particionamento do extrato bruto por extração líquido-líquido com hexano, acetato de etila e uma mistura etanol:água (1:2), dando origem as frações hexânica, acetato e hidroalcoólica, o que permitiu separar compostos por diferença de polaridade. Posteriormente, as três frações obtidas foram submetidas ao bioensaio, na concentração de 1mg/orelha, para determinar o grupo de maior atividade, sendo a fração hidroalcoólica a que apresentou maior atividade (43,6%). Posteriormente, as três frações obtidas foram submetidas ao bioensaio, na concentração de 1mg/orelha, para determinar o grupo de maior atividade, sendo a fração hidroalcoólica a que apresentou maior atividade (43,6%). Através desta fração, foi possível isolar (por LC-DAD utilizando uma coluna semi-preparativa) e identificar (por LC-MS) uma substância que apresenta o núcleo de uma apigenina ligada a uma pentose. Além do extrato bruto e das frações de Allophylus racemosus Sw. foi testada a atividade anti-inflamatória dos óleos essenciais, sendo que o mesmo apresentou resultados estatisticamente significativos frente aos controles negativo e positivo (indometacina e Acheflan®). A atividade anti-inflamatória dos óleos essenciais está relacionada, principalmente aos compostos majoritários, γ-muuroleno (30,66%), (E)–cariofileno (18,56%) e biciclogermacreno (17,38%), além do composto α-humuleno (2,33%), também encontrado no fitomedicamento Acheflan® e sendo o responsável, junto com (E)–cariofileno, pela atividade anti-inflamatória do fitomedicamento como relatado na literatura. / The nature always aroused in man a charming allure, not only by the resources available for their food and maintenance, but also for their main source of inspiration and learning. Allophylus racemosus Sw., objective of study, has no reports of its use, nor when it comes to scientific research data bases on studies of their pharmacological activities. It belongs to the Family Sapindaceae that comprises 140 genus and about 1900 species worldwide. Among them the Allophylus genus has about 250 species. Although a considerable number of species, there is a scant amount of research related to this genre, nor that kind. This plant was collected in the region of Minas Gerais Atlantic Forest near city Alfenas-MG. For the phytochemical study and anti-inflammatory activity, the crude ethanol extract of Allophylus racemosus Sw. leaves were obtained by extraction in ethanol and subjected to paw edema induced by carrageenan and ear edema test, which demonstrated a statistically significant anti-inflammatory activity compared to the negative and positive controls (indomethacin). After that, the crude extract there was partitioning by liquid-liquid extraction with hexane, ethyl acetate and a mixture ethanol: water (1: 2) to give three fractions allowing separating compounds according polarity. Subsequently, the three fractions obtained were subjected to bioassay to determine the group of higher activity. The hydroalcoholic fraction presented the highest activity(43,6%). Through this fraction could be isolated (by LC-DAD using a semi-preparative column) and identifying (LC-MS) a substance that has a core apigenin linked to a pentose. In addition to the crude extract and fractions of Allophylus racemosus Sw. Was tested for anti-inflammatory activity of essential oils, and it showed statistically significant (24.8% and 41.3% at concentrations of 0.25% and 25%, respectively) results compared to the negative and positive controls (indomethacin and Acheflan®). The anti-inflammatory activity of the essential oils is mainly related to the major compounds, γ-muurolene (30.66%), (E) -caryophyllene (18.56%) and bicyclogermacrene (17.38%), and the compound α -humulene (2.33%), also found in Acheflan® phytomedicine and being responsible for along with (E)-caryophyllene, the anti-inflammatory activity of phytomedicine as reported in the literature.

The volume homeostasis of retinal glial cells is mediated by an autocrine purinergic mechanism of ion channel opening which is activated in response to a decrease in the extracellular osmolarity. Here, I show that erythropoietin (EPO) prevents the osmotic swelling of glial somata in retinal slices from control and diabetic rats, with a half-maximal effect at approximately 0.01 nM. The downstream signaling evoked by EPO includes a release of vascular endothelial growth factor from the cells which was blocked by Janus kinase and extracellular signal-regulated kinases (ERK)1/2 inhibitors. Transactivation of kinase insert domain-containing receptor/fms-like tyrosine kinase 1 (KDR/flk-1) evokes a calcium-dependent, exocytotic release of glutamate, followed by activation of group I/II metabotropic glutamate receptors which results in calcium-independent release of ATP and adenosine from the cells. The final step in this cascade is the activation of adenosine A(1) receptors which results in protein kinase A- and phosphoinositide 3-kinase-mediated opening of potassium and chloride channels. EPO receptor protein was immunohistochemically localized to the inner retina and photoreceptor inner segments. In isolated glial cells, EPO receptor protein is selectively localized to fibers which traverse the inner nuclear layer in situ. Inhibition of glial swelling might contribute to the neuroprotective action of EPO in the retina under pathological conditions.

ObjetivosEstudiar el papel de la extracción de la membrana limitante interna (MLI) durante la vitrectomía posterior por vía pars plana (VPP), en el tratamiento quirúrgico de lo agujero macular (AM) y de lo edema macular crónico diabético (EMCD). Hemos estudiado las tasas de éxito clínico de la extracción de la MLI durante la VPP. Además hemos analizado el procedimiento técnico de extracción de la MLI, el aporte de la extracción de la MLI en la resolución clínica del EMCD después de la VPP y el valor de la tomografía de coherencia óptica (TCO) en el seguimiento de pacientes con EMCD y AM.El objetivo secundario consistía en establecer una correlación clínico-patológica mediante el examen histológico de la MLI. Este estudio abarcaba los descubrimientos histológicos y las diferencias de la MLI observadas mediante microscopia electrónica de transmisión (MET) y microscopia óptica (MO).Material y MétodosEstudio prospectivo de un año de 28 pacientes, no randomizado, no comparativo, con dos grupos de pacientes: 1) grupo con AM; 2) grupo con EMCD resistente al tratamiento láser.Los pacientes habían sido sometidos, antes y después de la VPP, a un examen oftalmológico: refracción objetiva, mejor agudeza visual (AV) corregida con teste de Snellen, medición de la presión intraocular, biomicroscopia, examen del fondo ocular y oftalmoscopia indirecta. El seguimiento postoperatorio fue efectuado durante el primer año a un día, una semana, 1, 3, 6 y 12 meses del postoperatorio. Los pacientes fueron sometidos a angiografía fluoresceínica digital, fotografía del fondo de ojo y mapado macular con TCO, como exámenes diagnósticos complementarios. Todos los pacientes fueron sometidos a la VPP y el objetivo principal era la disección epiretiniana y extracción de la MLI. Las MLIs fueron preparadas para un análisis con MO y MET. ResultadosEn el grupo con AM formado por diez ojos, seis tenían AM de tipo 2 y 3 y cuatro de tipo 4 (clasificación de Gass). Cinco pacientes presentan AM idiopáticos. Todos, salvo uno, mantuvieron o mejoraron la mejor AV corregida al final del estudio. La mejora de la VA era significativa estadísticamente sólo después del tercer mes del postoperatorio. El seguimiento mediante OCT demostró que el cierre del los AM fue alcanzado en nueve de ellos. La TCO pudo acompañar la evolución anatómica del cierre del agujero. En los 18 pacientes con EMCD, trece ojos mejoraron la AV, en dos no se produjo ningún cambio y tres empeoraron la AV, al final del seguimiento. La mejora de la AV comenzó a ser significativa entre el tercer y sexto mes del postoperatorio. El espesor foveal postoperatorio se redujo de manera significativa y progresiva respecto al observada en el preoperatorio. Hemos observado una disminución graduada y evidente del espesor macular paralelamente con una mejora progresiva de la AV: cuanto menos espessa era la fovea (TCO), mayor resultaba la mejor AV corregida. Hemos analizado las características ultraestructurales de la MLI normal en las retinas extraídas de dos donadores de ojos y comparado la MLI obtenida de los ojos sometidos a la VPP. Hemos realizado el examen histopatológico en 10 muestras del grupo con AM. Hemos podido observar la presencia de la MLI en 9 muestras. Hemos encontrado residuos de retina neurosensorial asociada con la superficie retinal de la MLI en 3 muestras. Pocas MLI presentaban un componente celular asociado con la superficie vítrea. En algunas MLI, hemos observado la presencia de fragmentos condensados de la hialoides posterior, a veces asociado con componente celular. Hemos llevado a cabo 75 mediciones correspondientes a distintos segmentos de la MLI. Los resultados han demostrado una significativa diferencia (p = 0.00011) entre el promedio del espesor de la MLI en el grupo con AM y en máculas normales. Al comparar el promedio de espesor observado en el grupo con AM con los obtenidos en el grupo con EMCD (2.2 ± 0.78 µm), la diferencia era siempre significativa (p = 0.03).En las muestras diabéticas examinadas mediante MET, hemos podido observar la presencia de la MLI en 7 muestras sobre 10. En 3 muestras sobre 10 no había ninguna MLI y la membrana epiretiniana presentaba una gran cantidad de matriz extracelular (colágeno y una gran variabilidad de células). El único descubrimiento histopatológico observado en 5 muestras mediante MET consistía en segmentos de MLI sin componentes fibrilares o celulares. Cuatro muestras presentaban una abundante disposición de colágeno compatible con el espesoramiento de la membrana hialoides o MER. Distintos tipos que rodeaban el colágeno extracelular componen la población celular asociada al tejido diabético: a) células glials; b) células fibroblásticas; c) células epiteloides; d) células plasmáticas macrófagas. En la MET hemos podido demostrar la presencia de componentes retinales neurosensoriales asociadas con la superficie retiniana de la MLI en 3 muestras.ConclusionesGrupo de Agujeros Maculares: 1. La VPP asociada a la extracción de la MLI es un procedimiento de utilidad para el tratamiento quirúrgico de los AM. En el presente estudio prospectivo de un año, la tasa de cierre anatómico fue de un 90% y la mejor media de la AV final corregida se obtuvo en los ojos con éxito anatómico.2. La TCO contribuyó para el diagnóstico, se presentó como una alternativa no invasiva para la evaluación del estado foveal y permitió documentar y analizar la evolución del cierre anatómico de los AM.Grupo de Edema Macular Diabético:1. El presente estudio clínico prospectivo de un año demostró los beneficios de la extracción de la MLI, mediante VPP, como tratamiento quirúrgico del EMCD. La extracción de la MLI conllevó a una disminución del engrosamiento de la retina, a la resolución clínica del edema macular y a una mejor AV.2. La TCO permitió un seguimiento cualitativo del perfil de la retina y un análisis cuantitativo de las pequeñas variaciones del espesor macular. El espesor de la macula ha disminuido significativamente al final del primer mes del postoperatorio (p = 0,00016) comparado con una mejoría significativa de la AV corregida a partir del sexto mes de seguimiento clínico (p = 0,05).3. Mediante MO y MET, el estudio histológico mostró la ausencia de fragmentos de MLI en algunos de los especimenes analizados. Algunas piezas histológicas eran membranas epirretinianas o la hialoides posterior engrosada. Ambos Grupos:1. La correlación clínico-patológica indicó que la presencia de elementos de la retina neurosensorial observados en la cara retiniana de algunos especimenes de MLI fue extraída de ojos que presentarían una peor AV final corregida.2. El espesor de la MLI observado en el grupo de AM fue mayor que el observado en el grupo de EMCD (p = 0,03) que, a su vez, fue mayor que el espesor observado en la MLI de la retina normal (p = 0,00003).

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.

The heart and microvasculature have characteristics of a complex adaptive system. Extreme challenges faced by these organ systems cause structural changes which lead to global adaptation. To assess the impact of myocardial interstitial edema on the mechanical properties of the left ventricle and the myocardial interstitium, we induced acute and chronic interstitial edema in dogs. With chronic edema, the primary form of collagen changed from type I to III and left ventricular chamber compliance significantly increased. The resulting functional adaptation allows the chronically edematous heart to maintain left ventricular chamber compliance when challenged with acute edema, thus, preserving cardiac function over a wide range of interstitial fluid pressures. To asses the effect of microvascular occlusions, we reintroduced the Pallid bat wing model and developed a novel mathematical model. We hypothesized that microvessels can switch from predominantly pressure-mediated to shear-mediated responses to ensure dilation during occlusions. Arterioles of unanesthetized Pallid bats were temporarily occluded upstream (n=8) and parallel (n=4) to vessels of interest (20-65 mm). In both cases, the vessels of interest rapidly dilated (36+24 %, 37+33 %), illustrating that they responded appropriately to either decreased pressure or increased shear stress. The model not only reproduced this switching behavior, but reveals its origin as the nonlinear shear-pressure-radius relationship. The properties of the heart and microvasculature were extended to characterize a “Research-Intensive Community” (RIC) model, to provide a feasible solution consistent with the Boyer Commission, to create a sustainable physiology research program. We developed and implemented the model with the aim of aligning diverse goals of participants while simultaneously optimizing research productivity. While the model radically increases the number of undergraduate students supported by a single faculty member, the inherent resilience and scalability of this complex adaptive system enables it to expand without formal institutionalization.

Stimulation of C-fiber sensory neurons innervating the respiratory tract with electricity or capsaicin leads to the liberation of substance P, CGRP and other neuropeptides from the nerve terminals. Substance P (SP) binds to the NK-receptors on the membrane of vascular endothelial cells and elicits neurogenic inflammatory responses. These inflammatory responses include plasma leakage and the subsequent edema formation (Lundberg and Saria, 1983¡FMcDonald et al., 1988). Evans blue is a hydrophilic dye and is often used as a tracer of plasma leakage due to its¡¦ high affinity to the plasma proteins. Plasma leakage causes Evans blue extravasates from the blood vessels and remains in the tissues. The more plasma leaks from the blood vessels, the more Evans blue will extravasate into the tissues. Measuring extravasated Evans blue dye that is extracted from tissues, is useful for evaluation of the amount of plasma leakage. Potassium channel openers can inhibit neurogenic plasma leakage in the airways and urinary bladder (Hollywood et al., 1998). Evans blue directly stimulates large-conductance Ca2+-activated K+ channels in cultured endothelial cells of human umbilical vein (Wu et al., 1999). This suggests that it may influence the permeability of the microvessels in vivo. A previous study shows that Evans blue dye blocks capsaicin-induced cough and bronchospasm in the guinea pig (Bolaer et al., 1995). We postulated that pretreatment with Evans blue may influence the extent of neurogenic inflammation in the rat airways induced by the application of either SP or capsaicin. India ink was used as a colloidal tracer dye to label the leaky vessels. The present study investigated whether different concentration of Evans blue (0, 3, 15 and 30 mg/ml/kg) pretreatment could affect the plasma leakage and edema formation in rat lower airways in response to intravenous injection of either SP or capsaicin. The amount of plasma leakage was expressed by the area density of India ink-labeled leaky blood vessels. We also investigated whether Evans blue influenced the ultra-structural change in tracheal serous cells induced by intravenous injection of SP. Our results showed that pretreatment with high concentration of Evans blue reduced more than seven tenths of the area density of plasma leakage in the trachea caused by SP application (P<0.01); reduced more than seven tenths in the left main bronchus (P<0.01) and reduced about seven tenths in the right main bronchus (P<0.01), compared to the control group that received saline prior to SP. However, no statistical significance was observed in edema ratio between any two groups (P>0.05). In the neurogenic inflammation of the airways caused by injection of capsaicin, pretreatment with high concentration of Evans blue reduced more than seven tenths of the area density of plasma leakage in the trachea (P<0.01); reduced more than seven tenths in the left main bronchus (P<0.01) and reduced about seven tenths in the right main bronchus (P<0.01), respectively, compared to the control group that received saline prior to capsaicin. Pretreatment with high concentration of Evans blue prior to capsaicin also reduced more than eight-tenth in edema ratio (P<0.01). In the ultra-structure change of serous cells and the stastical analysis of the number of active serous per 1000£gm2 of tracheal epithelium, Evans blue pretreatment prior to SP significantly reduced the number of active serous cells by seven tenths (P<0.01) as compared to control group that received saline prior to SP. Therefore, we concluded that pretreatment with high concentration of Evans blue exerted its¡¦ effect by opening large-conductance Ca2+-activated K+ channels and inhibited the plasma leakage induced by SP or capsaicin. But no significant inhibition was observed in edema formation induced by SP application. Low concentration of Evans blue might enhance the neurogenic inflammation of the airway. Under the observation with SEM, we found that SP activated serous cells in airway epithelium, and high concentration of Evans blue pretreatment lowered the secretory activity of serous cells. Therefore Evans blue might inhibit the activation of serous cells.

FUNDAMENTOS. En nuestro medio no conocemos los factores desencadenates de edema agudo de pulmón cardiogénico (EAPC)) y la tasa de mortalidad al año en pacientes ³70 años. METODOLOGIA. El presente trabajo es un estudio retrospectivo-descriptivo y longitudinal. Se incluyeron 35 casos de pacientes ³70 años con Diagnostico de Edema Agudo de Pulmón Cardiogénico, atendidos en la Unidad de Trauma Shock del Hospital Nacional Alberto Sabogal Sologuren entre el 1 de enero del 2001 y 31 de Junio del 2002 siendo seguidos por un año para conocer su mortalidad. Sus datos clínicos, y ecocardiográficos fueron obtenidos de sus historias clínicas.

Abuta grandifolia (Mart.) Sandwith y Mansoa alliacea (Lam.) A. H. Gentry son dos especies vegetales endémicas utilizadas en la medicina tradicional durante siglos por la población peruana para el tratamiento de problemas inflamatorios, dolores de cuerpo y reumatismo. Objetivos: Determinar el efecto antiinflamatorio del extracto hidroalcohólico de la corteza de Abuta grandifolia (Mart.) Sandwith y el efecto antiinflamatorio del extracto hidroalcohólico de las hojas de Mansoa alliacea (Lam.) A. H. Gentry. Materiales y Métodos: Los extractos hidroalcohólicos de ambas especies se obtuvieron con la corteza de A. grandifolia y las hojas de M. alliacea, las cuales fueron maceradas con etanol 96% y agua destilada (7:3). La Prueba Límite, test de toxicidad aguda (DL50), utilizó ratones hembras Mus musculus albinus balb/c adultas y se les administró por vía oral las diferentes dosis de los extractos (100, 1000, 2000 y 5000 mg/Kg). El test de Edema Pedal inducido por carragenina se realizó con ratas hembras cepa albina Sprague Dawley (150-200g). Éstas fueron agrupadas en 8 grupos (n=6) y tratadas por vía oral con los extractos de A. grandifolia (100, 300 y 600 mg/Kg), M. alliacea (25, 50 y 100 mg/Kg) y el fármaco standard (diclofenaco sódico 10 mg/mL) media hora antes de la aplicación del agente flogístico λ-carragenina. Resultados: DL50 para ambos extractos es mayor a 5000 mg/Kg. A. grandifolia, a las dosis de 600 y 300 mg/Kg, y M. alliacea, a las dosis de 100, 50 y 25 mg/Kg, fueron efectivas en reducir el edema pedal inducido por carragenina con porcentajes de eficacia antiinflamatoria de 45,74%, 30,88%, 20,91%, 29,94% y 19,80% respectivamente. Conclusiones: Los extractos hidroalcohólicos de la corteza de A. grandifolia a las dosis de 300 y 600 mg/Kg, y de las hojas de M. alliacea a las dosis de 25, 50 y 100 mg/Kg tienen efecto antiinflamatorio. / Abuta grandifolia (Mart.) Sandwith and Mansoa alliacea (Lam.) AH Gentry are two endemic species used in traditional medicine for centuries by the Peruvian population to treat inflammatory problems, body aches and rheumatism. Aim of the study: To determine the anti-inflammatory effect of Abuta grandifolia (Mart.) Sandwith hydroalcoholic extract of the bark and anti-inflammatory effect of Mansoa alliacea (Lam.) A.H. Gentry hydroalcoholic extract of the leaves. Materials and Methods: The hydroalcoholic extracts of both species were obtained from the bark of A. grandifolia and leaves of M. alliacea, which were macerated with 96% ethanol and distilled water (7: 3). Limit test, acute toxicity test (LD50) were carried out with Mus musculus albinus balb/c mice adults, they were orally administered with diferent doses of extracts (100, 1000, 2000 and 5000 mg/Kg). The Paw oedema induced by carrageenan test was carried out with Sprague Dawley albino female rats (150-200g). They were grouped into 8 groups (n = 6) and were treated orally with the extracts of A. grandifolia (100, 300 and 600 mg/Kg), M. alliacea (25, 50 and 100 mg/kg) and standard drug (diclofenac sodium 10mg/ml) half an hour before the application of the phlogistic agent λ-carrageenan. Results: LD50 for both extracts is greater than 5000 mg/Kg. A. grandifolia at doses of 600 and 300 mg/Kg and M. alliacea at doses of 100, 50 and 25 mg/kg, were effective in reducing carrageenan-induced paw oedema with efficacy anti-inflammatory percentages of 45,74%, 30,88%, 20,91%, 29,94% and 19,80% respectively. Conclusions: The A. grandifolia hydroalcoholic extract of the bark at doses of 300 and 600 mg/Kg, and M. alliacea hydroalcoholic extract of the leaves at doses of 25, 50 and 100 mg/kg have antiinflammatory effect. Key words: paw oedema, hydroalcoholic extract, carrageenan, Abuta grandifolia, Mansoa alliacea, anti-inflammatory.