Tempol attenuates atherosclerosis associated with metabolic syndrome via decreased vascular inflammation and NADPH-2 oxidase expression.

Oxidative stress is an important factor in the generation of vascular injury in atherosclerosis. Chronic administration of fructose in rodents is able to facilitate oxidative damage. In the present study we evaluated the role of Tempol, a superoxide dismutase mimetic, on the effect of high fructose intake in apolipoprotein E-deficient (ApoE-KO) mice. Rodents were fed with fructose overload (FF, 10% w/v) for 8 weeks and treated with Tempol 1 mg/kg/day the latest 4 weeks. Tempol revert the pro-oxidant effects caused by FF, diminished lipid peroxidation and impaired vascular NADPH oxidase system through the downregulation of p47phox expression in the vascular wall. Tempol inhibited the expression of vascular adhesion molecule 1 (VCAM-1) in aorta and reduced the development of atheroma plaques. Our results indicate that tempol attenuates oxidative stress by interfering with the correct assembly of Nox2 oxidase complex in the vascular wall and is able to reduce atherosclerosis. Thus tempol represents a potential therapeutic target for preventing risk factors associated with metabolic syndrome.

El síndrome de Rett. Estrategias actuales para el fisioterapeuta / The Rett syndrome. Current strategies for the physiotherapist

Introducción. El síndrome de Rett (SR) constituye una entidad única que suele producirse por una condición dominante ligada al cromosoma X, siendo más frecuente en la población femenina y produciendo un fenotipo clínico definido. Las características motoras que lo configuran requieren especial atención por parte de la fisioterapia, que junto con otras estrategias, tales como la musicoterapia, la hidroterapia y/o la terapia multisensorial, pretenden reducir y/o prevenir las deformidades ortopédicas, controlar los movimientos estereotipados de la mano y mantener las habilidades comunicacionales. Objetivos. Analizar y definir los estudios acerca de la intervención con terapias físicas y estrategias terapéuticas en el SR. Metodología. Se realizó una revisión bibliográfica centrada en la búsqueda de artículos en lengua inglesa relacionados con el tratamiento por medios físicos en el SR publicados entre 1986 y 2006; para ello se utilizaron las bases de datos The Cochrane Library, PEDro y Pubmed. Conclusiones. Los estudios consultados describen estrategias y terapias concretas, que favorecen la intervención terapéutica en estos pacientes

Introduction. The Rett Syndrome (RS) establishes a unique entity that used to arise by a dominant condition linked to the X chromosome, being more frequently in females and causing a specific clinical phenotype. The motor characteristics that form it, require special attention from the physiotherapist, that together with other strategies, like musictherapy, hydrotherapy and/or multi-sensorial therapy, try to decrease and/or prevent the orthopaedic deformities, to control the stereotyped movements of the hand and to maintain the communicational capacities. Objectives. To analyze and define the studies related to the intervention with physical therapies and therapeutic strategies in the RS. Methodology. A literature review was fulfilled and focused on the search of articles in English language related with the treatment by physical means in the RS published between 1986 and 2006; for this purpose, the following data bases were used: The Cochrane Library, PEDro and Pubmed. Conclusions. The studies describe strategies and concrete therapies that improve the therapeutic intervention in these patients

Crystalline polymorphism and molecular structure of sodium pravastatin.

In this work different crystallization processes of sodium pravastatin are explored and a new polymorph is obtained. The analytical results of powder X-ray diffraction (PXRD) and thermal analysis for this new polymorph indicate that it is different from the polymorphs previously reported. This new crystal form shows different physical-chemical properties than the previous forms, such as crystallographic structure, thermal behavior, and melting point, 181.5 degrees C. Besides, all crystallization processes previously reported use an aprotic solvent as antisolvent. However, we propose a new crystallization process for sodium pravastatin that uses only protic solvents, overcoming industrial scaling and environmental problems. Variable-temperature PXRD experiments show a transformation between different crystal forms in the range of 80-120 degrees C. Solid-state 13C NMR, reported in this work for the first time, and Fourier transform infrared (FT-IR) studies of some polymorphs show some differences in intermolecular interactions, especially with carboxylate and hydroxyl groups. Quantum mechanical calculations of the pravastatin molecule are also presented for the first time, obtaining a molecular structure similar to the experimental structure existing within the crystal lattice of the tert-octylamonium salt of pravastatin.

Changes in the elastase activity and colonization ability of Aspergillus fumigatus after successive inoculations in mice(AU)

En estudios previos en nuestro laboratorio hemos demostrado la existencia de una clara relación entre la actividad elastasa y la patogenicidad mediante el cálculo de lo que hemos denominado Índice de Actividad Elastasa (IAE). En el presente trabajo hemos evaluado la posibilidad de variación de este índice como consecuencia de inoculaciones sucesivas en ratones. Hemos utilizado dos cepas de Aspergillus fumigatus aisladas del ambiente que no presentaban actividad elastasa. Estas cepas se inocularon a grupos de diez ratones en sucesivos lotes. Nuestros resultados muestran que con cada inoculación se producía un incremento en el número de ratones de cada lote de los que se podía aislar la cepa fúngica, así como un incremento del número de aislados conun IAE>1. Esto nos sugiere que se produce una adaptación del hongo al medio en que se desarrolla y un incremento de su patogenicidad en su paso de hospedador a hospedador(AU)

In a previous work we demonstrated a clear link between elastase activity and pathogenicity using what we have named the Elastase Activity Index (EAI). In the present study we have evaluated the possible variability of this index as a consequence of successive inoculations in mice. Two strains of Aspergillus fumigatus isolated from the environment without elastase activity were used. These strains were inoculated into successive batches of ten mice. Our results showed that with each inoculation there was an increase in the number of mice on each batch from which the strain could be isolated and an increase in the number of strains with an EAI>1. This study suggests that A. fumigatus could adapt to the environment in which it is developed, increasing its pathogenic capabilities from host to host(AU)

Dehydroleucodine inhibits vascular smooth muscle cell proliferation in G2 phase.

Vascular smooth muscle cell (VSMC) proliferation plays an important role in the development of atherosclerosis and in the vascular changes seen in hypertension. Dehydroleucodine (DhL) is a sesquiterpene lactone that inhibits cell proliferation in plant cells. In this paper, we study the effect of DhL in the proliferation of VSMCs stimulated with 10% fetal bovine serum (FBS). Very low concentrations of DhL (2-6 microM) inhibited VSMC proliferation and induced cell accumulation in G2. DhL did not affect the dynamics of 3H-thymidine incorporation, and did not modify either the activity of DNA polymerase or the incorporation of deoxyribonucleotides in an in vitro assay. Moreover, DhL did not induce apoptosis in VSMCs. These results indicate that DhL, in very low concentration, induces a transient arrest of VSMCs in G2. Our data show that VSMCs are especially sensitive to DhL effect, suggesting that DhL could be potentially useful to prevent the vascular pathological changes seen in hypertension and other vascular diseases.

Effect of nebivolol on cardiovascular changes associated with a rat model of insulin-resistance.

Nebivolol is a vasodilator that combines beta-adrenergic blocking activity with a relaxant effect on vascular smooth muscle cells (VSMC) mediated by the endothelial nitric oxide (NO) pathway. FFR provide a model of dietary-induced insulin-resistance syndrome, which has been used to study the pathophysiological mechanisms associated with this syndrome. Our main objective was to examine the effect of long-term administration of nebivolol on metabolic and cardiovascular variables in fructose-fed rats (FFR), a model in which an altered bioavailability of NO has been already described. Male Wistar rats were randomly assigned to 4 groups (n = 8 each): I. Control (C); II. Control + nebivolol (C+N): 1 mg/kg(-1) x day(-1) in drinking water during the last 4 weeks. III. FFR: rats receiving fructose in drinking water as a 10% (w/v) solution during 8 weeks, and IV. FFR+N: idem II plus III. During the 8 weeks experimental period, variations in systolic blood pressure (SBP), glucose tolerance test (GTT) and plasma thiobarbituric acid-reactive substances (TBARS) were assessed. At the end of this experimental period, rats were killed and heart and kidneys were excised for calculation of relative heart weight (RHW) and histological evaluation of lumen to media ratio (L/M) in renal arteries. Rats from FFR group increased their SBP and RHW, showed glucose intolerance and an increment in lipid peroxidation. Moreover, FFR showed vascular remodeling in renal arteries evidenced by changes in L/M. Although the metabolic changes were not reverted by the administration of nebivolol, this drug successfully decreased SBP, TBARS levels and reverted structural changes such as cardiac hypertrophy and renal arterial remodeling. Data demonstrate that nebivolol administration could participate in the reversion of cardiovascular structural changes associated with the insulin-resistance syndrome.

Chronic administration of losartan reverses cardiovascular changes in hypertensive fructose-fed rats.

The cluster of risk factors including hyperinsulinemia, insulin resistance, hypertriglyceridemia and hypertension has been called syndrome X. Several evidences link the insulin resistance syndrome with endothelial dysfunction. Since the participation of the renin-angiotensin system (RAS) in this pathology is still unclear, the present study examined the effect of chronic administration of an angiotensin AT1 receptor antagonist, losartan (L), on endothelial nitric oxide synthase (eNOS) activity in aortic endothelium and cardiac tissue, and on the proliferation of primary cultured aortic smooth muscle cells (SMC), obtained from fructose-fed rats (FFR), an experimental model of syndrome X Male Wistar rats were used: Control, FFR and FFR+L (n = 8 in each group). After 8 weeks, tissue samples were obtained and 10% fetal calf serum (FCS) proliferative effect was examined in SMC by 3H-thymidine incorporation and cell counting. The eNOS activity was estimated in aortic endothelial lining and cardiac homogenates by conversion of 3H-arginine into 3H-citrulline. FFR aortic SMC showed a significantly increased 10% FCS-induced 3H-thymidine incorporation and cell number compared to controls. FFR aortic and cardiac eNOS activities were significantly decreased. Chronic treatment with L decreased systolic blood pressure,reverted cardiac hypertrophy, abolished the increased SMC proliferation and restoredeNOS activity. These data confirm that changes in SMC proliferation and endothelial dysfunction at different levels of the cardiovascular system are involved in syndrome "X", and that AT1 receptor blocking can revert those changes, suggesting an important role of the RAS, possibly mediated by AT2 receptors and kinins, in the physiopathological mechanisms of this model.

Body composition and vascular effects of growth hormone administration in old female rats.

Aging is associated with alterations in cardiovascular system and changes in body composition. The aim of this study was to investigate the effect of GH on body composition, vascular function and structure in old female rats. Old (20 months) and adult (4 months) female Wistar rats were used. One group of old animals was treated with GH (2 mg/kg/day) for four weeks. Periuterine fat weight, specific gravity index (SGI), dose response to Acetylcholine, Isoprenaline, Phenylephrine and Acetylcholine in the presence of L-NAME and vascular morphology in aortic rings, were studied. Old rats showed increased fat weight and decreased SGI (p<0.05) as compared to adult animals. GH reduced fat weight (p<0.05) and tended to increase SGI (NS). Old rats showed impaired vasodilatation to Acetylcholine and Isoprenaline (p<0.05), and GH improved these responses (p<0.05). Contraction response to Phenylephrine was higher in old than in adults rats (p<0.05), but GH did not show any effect. Contraction induced by Acetylcholine+L-NAME was higher in old rats than in adults, and GH tended to reduce this response, although not significantly. Aortic media area was increased in old rats, and GH reduced this parameter (p<0.05). In conclusion, GH shows beneficial effects on body composition, vascular function and morphology in old female rats.

Proteoglycans production by aortic vascular smooth muscle cells from hypertensive rats

Remodeling of large and small arteries contributes to the development and complications of hypertension. Artery structural changes in chronic sustained hypertension include vascular smooth muscle cells (VSMC) proliferation and extracellular matrix (ECM) modifications. Extracellular constituents such as proteoglycans (PGs), may modulate vascular stiffness and VSMC growth and differentiation. We examined the effect of growth factors on secreted and membrane-bound PGs synthesis by cultured aortic smooth muscle cells (SMC) from 12- to 14- week-old spontaneously hypertensive rats (SHR) and age-matched Wistar rats. After stimulation with platelet-derived growth factor (PDGF-BB), 10% fetal calf serum (FCS) or 0.1% FCS as control, PGs synthesis (dpm/ng DNA) was evaluated in the medium (M-ECM) and in the cell layer (P-ECM) by a double-isotopic label method using both [3H]-glucosamine and [35S]-sodium sulfate which are incorporated into all complex carbohydrates or only into sulfated dysaccharides, respectively. Data are presented as percent of the control (0.1% FCS). SHR VSMC displayed a significantly greater synthesis of M-ECM [3H]-PGs than Wistar rat cells, with both treatments, but no differences in M-ECM [35S] uptake were found in any case. In the P-ECM, both PDGF-BB and 10% FCS produced a greater effect on [3H]-PGs and sulfated PGs synthesis in VSMC from SHR. An important change seen in SHR cells was a significant decreased sulfation, assessed by [35S]/[3H] ratio, in basal and stimulation conditions. Present results indicate the existence of changes in PGS synthesis and modulation in VSMC from a conduit-artery of SHR and support the pathophysiological role proposed for matrix proteoglycans in the vascular wall changes associated to hypertension and related vascular diseases as atherosclerosis.

Proteoglycans production by aortic vascular smooth muscle cells from hypertensive rats

Remodeling of large and small arteries contributes to the development and complications of hypertension. Artery structural changes in chronic sustained hypertension include vascular smooth muscle cells (VSMC) proliferation and extracellular matrix (ECM) modifications. Extracellular constituents such as proteoglycans (PGs), may modulate vascular stiffness and VSMC growth and differentiation. We examined the effect of growth factors on secreted and membrane-bound PGs synthesis by cultured aortic smooth muscle cells (SMC) from 12- to 14- week-old spontaneously hypertensive rats (SHR) and age-matched Wistar rats. After stimulation with platelet-derived growth factor (PDGF-BB), 10% fetal calf serum (FCS) or 0.1% FCS as control, PGs synthesis (dpm/ng DNA) was evaluated in the medium (M-ECM) and in the cell layer (P-ECM) by a double-isotopic label method using both [3H]-glucosamine and [35S]-sodium sulfate which are incorporated into all complex carbohydrates or only into sulfated dysaccharides, respectively. Data are presented as percent of the control (0.1% FCS). SHR VSMC displayed a significantly greater synthesis of M-ECM [3H]-PGs than Wistar rat cells, with both treatments, but no differences in M-ECM [35S] uptake were found in any case. In the P-ECM, both PDGF-BB and 10% FCS produced a greater effect on [3H]-PGs and sulfated PGs synthesis in VSMC from SHR. An important change seen in SHR cells was a significant decreased sulfation, assessed by [35S]/[3H] ratio, in basal and stimulation conditions. Present results indicate the existence of changes in PGS synthesis and modulation in VSMC from a conduit-artery of SHR and support the pathophysiological role proposed for matrix proteoglycans in the vascular wall changes associated to hypertension and related vascular diseases as atherosclerosis. (AU)

Aortic smooth muscle cell proliferation and endothelial nitric oxide synthase activity in fructose-fed rats.

The aim of this study was to evaluate the proliferative behavior of vascular smooth muscle cells in primary culture (pC-SMC) and the endothelial nitric oxide synthase (eNOS) activity in the endothelial lining of the aorta of fructose-fed rats (FFR). This is an experimental model of syndrome X, a cluster of cardiovascular risk factors including hyperinsulinemia, insulin resistance, and hypertension that has been suggested to be of pathophysiologic importance for the development of atherosclerosis. Male Wistar rats were used: Control (n = 12) and FFR (n = 12). After receiving fructose in drinking water (10% w/v) during 8 weeks, biochemical parameters, systolic blood pressure (SBP) and relative heart weight (RHW) were determined. The proliferative effect of 10% fetal calf serum (FCS) was examined in aortic pC-SMC by [3H]thymidine incorporation and by cell counting. Ca2+/calmodulin-dependent NOS activity was estimated in aortic endothelial lining and in heart tissue homogenates by conversion of [3H]arginine into [3H]citrulline. Fructose-fed rats showed hyperinsulinemia (P = .0263), altered glucose tolerance test (P < .001), higher SBP (P < .0001), and RHW (P = .0145), compared to control rats. These animals also showed an increase of 10% FCS-induced [3H]thymidine incorporation (P < .0001) and cell number of aortic pC-SMC (P = .0049) and decreased eNOS activity in both aortic endothelium (P = .0147) and cardiac tissue (P < .0001). These data support the hypothesis that syndrome X is associated to changes in SMC proliferation and endothelial dysfunction, which could be involved in the onset or progression of the atherogenic process.

Un modelo de gestión de la calidad total para la enfermería hospitalaria / A model of total quality management for the hospital nursing

Introducción: El modelo Europeo para la Gestión de la calidad Total (EFQM) se sintetiza de la siguiente manera: La satisfacción de los clientes y empleados, se consiguen mediante un liderazgo que impulse la política y estrategia de la organización a través de una adecuada utilización de los recursos con una perfecta gestión de los procesos más importantes de la organización con objeto de conseguir unos resultados excelentes. Existen algunas experiencias aisladas en Europa y algunos proyectos en marcha en España que intenta adaptar el modelo EFQM para instituciones sanitarias. Este modelo, al igual que puede ser aplicable al contexto general de las actividades que se llevan a cabo en un hospital, también sería posible circunscribirlo al área de los cuidados de enfermería. Es precisamente ésta aplicación la que hemos llevado a cabo. Objetivo: Evaluar mediante un modelo de excelencia de gestión de calidad total la enfermería hospitalaria. Material y Métodos: Se presenta un estudio basando en técnicas de investigación cualitativas (brainstorming, Técnica de grupo nominal-TGN), en el que diferentes enfermeros expertos (clínicos, gestores y docentes) y pacientes lograron consenso sobre los aspectos, contenidos, instrumentos y métodos, que basados en los criterios del modelo EFQM, sirvan para medir la calidad de los cuidados de enfermería en los hospitales. Completado el modelo, que consta de más de 500 áreas o criterios de evaluación, se aplicó en el área materno infantil de 3 hospitales del Servicio Andaluz de Salud. Resultados: Se contrastan los resultados obtenidos en los tres centros y las dificultades que ha presentado su aplicación. De los resultados se desprende que en todos los criterios y en todos los hospitales existen innumerables aéreas que mejorar y que las puntuaciones obtenidas (373,4, 174,3 y 193,1 sobre 1.000) distan mucho de las obtenidas por organizaciones excelentes. Conclusión: El modelo nos muestra una herramienta útil y exhaustiva, que puede ser utilizada para la detección y priorización de oportunidades de mejora, sobre las cuales podemos intervenir para alcanzar mejoras en las distintas áreas abordadas (AU)

The value of the determination of anti-Aspergillus IgG in the serodiagnosis of canine aspergillosis: comparison with galactomannan detection.

Diagnosis of canine aspergillosis is difficult using currently available methods. It often passes unnoticed or is diagnosed in the later phases of the disease. We developed an ELISA technique to detect anti-Aspergillus antibodies in canine serum using an Aspergillus antigenic mycelial extract, which could then be used for the diagnosis of canine aspergillosis. We used a cut-off of X + 3SD obtained from 20 control sera. The test was performed on 46 dogs with lesions indicating possible aspergillosis and gave nine positive results: one systemic mycosis, two discospondylitis, one uveitis, two bronchopulmonary processes and three rhinitis. We compared this methodology with the PLATELIA technique in the follow-up of the affected dogs, obtaining the same limitations as in the diagnosis of human aspergillosis. We consider our ELISA technique using sera samples a speedy, safe and reliable method which enables us to follow up the evolution of the disease and the efficacy of the therapy chosen. A definitive diagnosis must still take into account the results of other tests such as clinical examination, radiographic studies, endoscopy and biopsy.

Proteoglycan production by vascular smooth muscle cells from resistance arteries of hypertensive rats.

Extracellular matrix (ECM) modifications in the vascular wall contribute to the narrowing of arteries in hypertension. Because direct evidence for the role of proteoglycans (PGs) in the pathological process of resistance-sized arteries has not already been demonstrated, we examined the effect of growth factors on secreted and membrane-bound PG synthesis by cultured mesenteric vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and Wistar rats. After 48 hours of stimulation with angiotensin II (Ang II), platelet-derived growth factor (PDGF-BB), and 10% fetal calf serum (FCS) or 0.1% FCS as control, PG synthesis (in dpm/ng DNA) was evaluated in the medium (M-ECM) and in the cell layer (P-ECM) by a double-isotopic label method with both [(3)H]-glucosamine and [(35)S]-sodium sulfate, which are incorporated into all complex carbohydrates or only into sulfated disaccharides, respectively. VSMC from SHR displayed a significantly lower level of synthesis of M-ECM [(3)H]-PGs than those of Wistar rats in all the experimental groups, including the control group (0. 1% FCS), but no differences in M-ECM [(35)S] uptake were found in any case. In the P-ECM, Ang II was the only factor that produced a lesser effect on [(3)H]-glucosamine and a greater effect on [(35)S]-sodium sulfate uptakes in VSMC from SHR than from Wistar rats. The most prominent change seen in VSMC from SHR was an increased sulfation, assessed by [(35)S]/[(3)H] ratio, in nonstimulated cells and in response to 10% FCS and Ang II but not to PDGF-BB compared with VSMC from Wistar rats. These data indicate the existence of changes in PG modulation in the resistance vessels of SHR, which suggests that PGs may contribute to the development of structural and functional modifications in hypertensive states.

[Accumulation of actin and adhesion to HEp-2 cells of strains of Escherichia coli isolated from children with diarrhea in Mendoza, Argentina]. / Acumulación de actina y adherencia a células HEp-2 de cepas de Escherichia coli aisladas de niños con diarrea en Mendoza, Argentina.

E. coli strains are the major bacterial cause of diarrhea among children under 2 years of age residing in Mendoza, Argentina. Detection of diarrheogenic E. coli is made after coproculture, by agglutination tests using O-group antisera including most enteropathogenic E. coli (EPEC) serogroups and others often isolated in diarrhea. Although there are many O serogroups and H serotypes of E. coli strongly associated with infantile diarrhea, a number of studies have shown differences in the rate of isolation of EPEC between cases and controls using DNA probes. We compared the diagnosis of EPEC infections by traditional serogrouping tests with other detection methods using cell culture, involving the screening of isolates for adherence patterns to HEp-2 cells. A total of 140 isolates from children less than 24 months old with acute, persistent and chronic diarrhea and 40 isolates from controls were recovered. Three distinct patterns of adherence, termed localised (LA), diffuse (DA) and aggregative (EAgg) adherence were found. The fluorescence actin staining assay was used as indicative of the ability of some EPEC strains that produce attaching-effacing (A/E) lesions. Positive serogrouping strains were strongly associated with adherence (P = 0.0001). LA adherence pattern occurred in 11% of cases with acute diarrhea associated with these serogroups (P = 0.001) and children under 12 months (P = 0.0001). The FAS test was positive in 80% of them. EAgg adherence was found only in patients (20% P = 0.0001) and DA occurred both in cases (29%) and controls (2.5% P = 0.0001). Diagnosis of EPEC infections has traditionally been performed by identifying organisms belonging to a number of serogroups or serotypes epidemiologically linked to diarrhea. Evidence is presented in this paper to show that pathogenicity is not restricted to serogroups. Isolation of many adherent strains not belonging to traditional EPEC O serogroups, shows the need for alternative methods to be used to detect and identify E. coli.

Proliferative effect of insulin on cultured smooth muscle cells from rat mesenteric resistance vessels.

The present study evaluates the growth promoting effect of insulin on the proliferative activity of cells from rat mesenteric arteries in culture in order to test the hypothesis that insulin may play a pathogenetic role in the hypertrophy of resistance vessels. The proliferative effect of insulin was studied in cultured vascular smooth muscle cells (SMC) obtained from two functionally different rat vessels: mesenteric arteries and aorta. Growth characteristics (cell number and growth rate) of mesenteric and aortic cells were determined after a quiescent period and followed-up for 24, 72, and 120 h after the addition of insulin. At all studied time intervals, aortic SMC exhibited a significatively higher cell number and specific growth rate than did mesenteric SMC. Aortic SMC also displayed a greater proliferation than did mesenteric SMC in the presence of 10% fetal calf serum (FCS). At a physiological concentration of 100 microU/mL, the proliferative effect of insulin, after a quiescent period, was seen only in aortic SMC, at 72 and 120 h. Higher insulin concentration (500 microU/mL) increased significantly the cell number in SMC of both arteries. The proliferative effect was significant at all studied periods for aortic SMC; however, in mesenteric SMC, insulin increased the cell number only at 72 and 120 h. The proliferative effect of insulin was observed on SMC obtained from functionally different arteries such as aorta and mesenteric, being greater in the former. The different behavior of these SMC in the presence not only of insulin, but also of 10% FCS, provides further evidence for the existence of intervascular heterogeneity. The mild stimulatory effect of insulin in vitro may contribute in this way to the vascular hypertrophy of pathological entities exhibiting hyperinsulinemia.

Acumulación de actina y adherencia a células HEp-2 de cepas de Escherichia coli aisladas de niños con diarrea en Mendoza, Argentina / Actin accumulation and HEp-2 cell adherence assay for Escherichia coli isolated from children in Mendoza, Argentina

Entre los niños menores de 2 años atendidos en el Hospital Infantil H. Notti de Mendoza, Argentina, es frecuente la diarrea causada por E. coli. La identificación se realiza por medio de marcadores bioquímicos y seroagrupamiento con antisueros que incluyen la mayoría de los serogrupos tradicionales de E. coli enteropatógeno (EPEC). Aunque se ha comprobado que existen serogrupos O y serotipos H de E. coli comúnmente asociados a diarrea infantil, en algunos estudios existe discrepancia tanto en relación a su presencia en controles sanos, en cuanto a su clasificación como EPEC en estudios genotípicos con sondas de ADN. En este trabajo se comparó la información proporcionada por la metodología estándar con los patrones de adherencia a células HEp-2, de cepas obtenidas de coprocultivos de 140 niños menores de 2 años con diarrea aguda, prolongada y crónica. Se hizo un grupo control de 40 casos sin diarrea. Las cepas se clasificaron según serología positiva (+) y negativa (-) frente a antisueros polivalentes. Se identificaron 3 tipos de adherencia: localizada (LA), difusa (DA) y EAagg (agregativa). Además, se realizó la prueba denominada "Fluorescence Actin Staining" (FAS) que pone en evidencia la lesión característica de EPEC, denominada "attaching-effacing" (A/E) producida por la formación de un pedestal de filamentos de actina en el sitio de adhesión bacteriana, en tejido intestinal y células en cultivo. En diarreas agudas, la adherencia fue relacionada con la serología + (P=0,001). La presencia de LA en diarreas fue del 11 por ciento con predominio significativo en niños menores de 12 meses (P=0,0001) y serología + (P=0,0001). El 80 por ciento de estas cepas dieron positiva la prueba FAS. El patrón DA apareció en diarreas (29 por ciento P=0,0001) y controles (2,5 por ciento); EAgg sólo en diarreas (20 por ciento P=0,0001). Este trabajo postula que la patogenicidad no estaría restringida a serogrupos. Debido a la frecuencia del aislamiento de cepas enteroadherentes de las que no se conoce bien la patogenia y que no son detectadas por las pruebas fenotípicas estándares, sería importante complementar al laboratorio clínico con técnicas que evidencien factores de virulencia

Acumulación de actina y adherencia a células HEp-2 de cepas de Escherichia coli aisladas de niños con diarrea en Mendoza, Argentina / Actin accumulation and HEp-2 cell adherence assay for Escherichia coli isolated from children in Mendoza, Argentina

Entre los niños menores de 2 años atendidos en el Hospital Infantil H. Notti de Mendoza, Argentina, es frecuente la diarrea causada por E. coli. La identificación se realiza por medio de marcadores bioquímicos y seroagrupamiento con antisueros que incluyen la mayoría de los serogrupos tradicionales de E. coli enteropatógeno (EPEC). Aunque se ha comprobado que existen serogrupos O y serotipos H de E. coli comúnmente asociados a diarrea infantil, en algunos estudios existe discrepancia tanto en relación a su presencia en controles sanos, en cuanto a su clasificación como EPEC en estudios genotípicos con sondas de ADN. En este trabajo se comparó la información proporcionada por la metodología estándar con los patrones de adherencia a células HEp-2, de cepas obtenidas de coprocultivos de 140 niños menores de 2 años con diarrea aguda, prolongada y crónica. Se hizo un grupo control de 40 casos sin diarrea. Las cepas se clasificaron según serología positiva (+) y negativa (-) frente a antisueros polivalentes. Se identificaron 3 tipos de adherencia: localizada (LA), difusa (DA) y EAagg (agregativa). Además, se realizó la prueba denominada "Fluorescence Actin Staining" (FAS) que pone en evidencia la lesión característica de EPEC, denominada "attaching-effacing" (A/E) producida por la formación de un pedestal de filamentos de actina en el sitio de adhesión bacteriana, en tejido intestinal y células en cultivo. En diarreas agudas, la adherencia fue relacionada con la serología + (P=0,001). La presencia de LA en diarreas fue del 11 por ciento con predominio significativo en niños menores de 12 meses (P=0,0001) y serología + (P=0,0001). El 80 por ciento de estas cepas dieron positiva la prueba FAS. El patrón DA apareció en diarreas (29 por ciento P=0,0001) y controles (2,5 por ciento); EAgg sólo en diarreas (20 por ciento P=0,0001). Este trabajo postula que la patogenicidad no estaría restringida a serogrupos. Debido a la frecuencia del aislamiento de cepas enteroadherentes de las que no se conoce bien la patogenia y que no son detectadas por las pruebas fenotípicas estándares, sería importante complementar al laboratorio clínico con técnicas que evidencien factores de virulencia (AU)

Renal functional reserve and microalbuminuria excretion in vesicoureteral reflux after surgery correction.

The acute effect of an oral protein load on glomerular filtration rate-renal functional reserve (RFR)- and albumin urinary excretion were evaluated in 9 patients with bilateral vesicoureteral grade IV reflux after surgical correction (Group I) and the results compared with 9 children with repeated urinary infection without reflux (Group II) and 6 healthy controls of similar age (Group III). Intravenous urography, performed in the year of the study, revealed renal scarring in 10 kidneys in Group 1 and 2 in Group II. All of them had normal values of plasma creatinine. Basal inulin clearance was significantly lower in vesicoureteral reflux patients. Good correlation was found between parenchymal area of both kidneys and baseline inulin. Following protein load an increase in creatinine and inulin clearance was recorded in urinary infection and control children. No change post load was observed in reflux patients. Microalbuminuria excretion was significantly higher in children with surgical correction during the control period. No changes were observed after load in any of the groups. We conclude that patients who had had bilateral vesicoureteral reflux grade IV showed an impaired renal response to a protein load. Long-term follow-up studies will confirm the value of this test for estimating the risk of further renal function deterioration in reflux nephropathy.

Renal functional reserve and microalbuminuria excretion in vesicoureteral reflux after surgery correction.

The acute effect of an oral protein load on glomerular filtration rate-renal functional reserve (RFR)- and albumin urinary excretion were evaluated in 9 patients with bilateral vesicoureteral grade IV reflux after surgical correction (Group I) and the results compared with 9 children with repeated urinary infection without reflux (Group II) and 6 healthy controls of similar age (Group III). Intravenous urography, performed in the year of the study, revealed renal scarring in 10 kidneys in Group 1 and 2 in Group II. All of them had normal values of plasma creatinine. Basal inulin clearance was significantly lower in vesicoureteral reflux patients. Good correlation was found between parenchymal area of both kidneys and baseline inulin. Following protein load an increase in creatinine and inulin clearance was recorded in urinary infection and control children. No change post load was observed in reflux patients. Microalbuminuria excretion was significantly higher in children with surgical correction during the control period. No changes were observed after load in any of the groups. We conclude that patients who had had bilateral vesicoureteral reflux grade IV showed an impaired renal response to a protein load. Long-term follow-up studies will confirm the value of this test for estimating the risk of further renal function deterioration in reflux nephropathy.