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BACKGROUND: Exposure of infant animals to clinically used anaesthetics is associated with acute structural brain abnormalities and development functional alterations. The α 2 -adrenoceptor agonist dexmedetomidine (DEX) induces sedation, analgesia, and provides neuroprotection in experimental brain injury models. However, it is unknown whether DEX also affords protection in the developing brain against anaesthesia using sevoflurane (SEVO), which is commonly used in paediatric anaesthesia. METHODS: Infant rats were exposed on postnatal day seven for six h to 2.5% SEVO and were given i.p. injections of saline or DEX (1-50 µg kg -1 ) three times during the exposure. Level of anaesthesia, respiratory rates, and arterial blood gasses were assessed for each animal. Apoptosis was determined in brain slices immunostained for activated caspase-3 (AC-3) using a computerised approach. RESULTS: SEVO alone induced a surgical plane of anaesthesia, and all animals survived the study. SEVO induced an approximately 10-fold increase in AC-3 positive cells in several cortical and subcortical brain regions compared with untreated control animals. Co-administration of DEX 1 µg kg -1 with SEVO significantly reduced apoptosis in all brain areas, affording the highest protection in the thalamus (84% reduction) and lowest in the hippocampus and cortical areas (â¼50% reduction). DEX 5-25 µg kg -1 plus SEVO dose-dependently increased infant rat mortality. CONCLUSIONS: SEVO anaesthesia induced widespread apoptosis in infant rat brain. Co-administration of DEX (1 µg kg -1 ) provided significant protection, whereas DEX (5 µg kg -1 or higher) plus SEVO increased mortality. Our findings suggest that DEX could be an attractive therapeutic for future studies investigating its neuroprotective potential in a translational animal model.
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Encéfalo/efectos de los fármacos , Dexmedetomidina/farmacología , Neuroprotección/efectos de los fármacos , Síndromes de Neurotoxicidad/prevención & control , Sevoflurano/efectos adversos , Anestésicos por Inhalación/efectos adversos , Animales , Animales Recién Nacidos , Apoptosis/efectos de los fármacos , Modelos Animales de Enfermedad , Hipnóticos y Sedantes/farmacología , Ratas , Ratas WistarRESUMEN
Quantum entanglement became essential in understanding the non-locality of quantum mechanics. In optics, this non-locality can be demonstrated on impressively large length scales, as photons travel with the speed of light and interact only weakly with their environment. Spontaneous parametric down-conversion (SPDC) in nonlinear crystals provides an efficient source for entangled photon pairs, so-called biphotons. However, SPDC can also be implemented in nonlinear arrays of evanescently coupled waveguides which allows the generation and the investigation of correlated quantum walks of such biphotons in an integrated device. Here, we analytically and experimentally demonstrate that the biphoton degrees of freedom are entailed in an additional dimension, therefore the SPDC and the subsequent quantum random walk in one-dimensional arrays can be simulated through classical optical beam propagation in a two-dimensional photonic lattice. Thereby, the output intensity images directly represent the biphoton correlations and exhibit a clear violation of a Bell-like inequality.
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RATIONALE: The MARAN (Macular Relocation in Age-related Neovascular disease) trial was planned to assess the effectiveness of full macular relocation (MR) in patients with neovascular age-related macular degeneration (AMD). DESIGN: Randomised, prospective, controlled clinical trial. METHODS: Patients suffering from visual loss because of AMD were randomised to either surgery or a control group receiving standard treatment (observation or photodynamic therapy (PDT)). The primary end point was the change of visual acuity (VA) (ETDRS) 52 weeks after randomisation compared with initial VA, and secondary end points included reading performance, contrast sensitivity, stability of fixation, eye-specific quality of life, and the absolute number of letters read correctly at 52 weeks compared with initial examination. RESULTS: Owing to early determination, only 28 patients were included in the study. The study did not show a difference between the two groups with respect to the final visual result or any of the secondary outcomes measured. The study was limited by the low recruitment that was, at least in part, attributed to the inherent risks for those patients randomised to the surgical arm of the study as well as to the emerging new treatments for AMD. CONCLUSION: The results of the MARAN trial failed to recruit a sufficient number of patients and a superiority of surgery over observation or PDT in patients with exudative AMD was not shown. There was a trend that the reading function was superior after surgery. In the light of the new pharmacological treatments, surgical options such as MR will be an option for only selected cases.
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Neovascularización Coroidal/cirugía , Degeneración Macular/cirugía , Anciano , Anciano de 80 o más Años , Neovascularización Coroidal/etiología , Neovascularización Coroidal/fisiopatología , Sensibilidad de Contraste/fisiología , Femenino , Fijación Ocular/fisiología , Humanos , Degeneración Macular/complicaciones , Degeneración Macular/fisiopatología , Masculino , Estudios Prospectivos , Calidad de Vida , Lectura , Análisis y Desempeño de Tareas , Agudeza VisualRESUMEN
Coordination Centres for Clinical Trials (Koordinierungszentren für Klinische Studien, KKS) were set up to increase the quality and number of clinical trials in Germany as well as to establish clinical trial training programs in order to improve international recognition of German clinical research. Over the past 6 years, 12 KKS have been set up at the respective universities with a public grant from the Federal Ministry of Education and Research (BMBF). Many non-clinical services have been established to ensure successful co-operation in clinical trials with clinical scientists and industry. KKS help researchers to efficiently conduct commercial and non-commercial clinical trials in various disease areas. Their expertise and infrastructure allow the university to assume sponsor responsibility in non-commercial drug trials. Because of their professional work and education activities KKS are well accepted by industry and the scientific community. Central professional trial organisations such as the KKS have been shown to be the pre-requisite for meeting the growing, manifold and complex requirements for clinical trials. Therefore they are considered essential for progress and success in clinical research.
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Estudios Multicéntricos como Asunto/legislación & jurisprudencia , Ensayos Clínicos Controlados Aleatorios como Asunto/legislación & jurisprudencia , Centros Médicos Académicos/legislación & jurisprudencia , Centros Médicos Académicos/organización & administración , Comités de Monitoreo de Datos de Ensayos Clínicos/legislación & jurisprudencia , Comités de Monitoreo de Datos de Ensayos Clínicos/organización & administración , Industria Farmacéutica/legislación & jurisprudencia , Alemania , Humanos , Comunicación Interdisciplinaria , Apoyo a la Investigación como Asunto/legislación & jurisprudencia , Apoyo a la Investigación como Asunto/estadística & datos numéricosRESUMEN
Despite the existence of many single ion sorption studies on iron and aluminum oxides, fewer studies have been reported that describe cosorption reactions. In this work, we present an in situ ATR FTIR study of synergistic adsorption of sulfate (SO4) and copper (Cu) on goethite, which is representative of the minerals and ions present in mine wastes, acid sulfate soils, and other industrial and agricultural settings. Sulfate adsorption was studied as a function of varying pH, and as a function of increasing concentration in the absence and presence of Cu. The presence of Cu ions in solution had a complex effect on the ability of SO4 ions to be retained on the goethite surface with increasing pH, with complete desorption occurring near pH 7 and 9 in the absence and presence of Cu, respectively. In addition, Cu ions altered the balance of inner vs outer sphere adsorbed SO4. The solid phase partitioning of SO4 at pH 3 and pH 5 was elevated by the presence of Cu; in both cases Cu increased the affinity of SO4 for the goethite surface. Complementary ex situ sorption edge studies of Cu on goethite in the absence and presence of SO4 revealed that the Cu adsorption edge shifted to lower pH (6.3 --> 5.6) in the presence of SO4, consistent with a decrease of the electrostatic repulsion between the goethite surface and adsorbing Cu. Based on the ATR FTIR and bulk sorption data we surmise that the cosorption products of SO4 and Cu at the goethite-water interface were not in the nature of ternary complexes under the conditions studied here. This information is critical for the evaluation of the onset of surface precipitates of copper-hydroxy sulfates as a function of pH and solution concentration.
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Cobre/química , Compuestos de Hierro/química , Sulfatos/química , Adsorción/efectos de los fármacos , Cobre/farmacología , Contaminantes Ambientales/química , Concentración de Iones de Hidrógeno/efectos de los fármacos , Iones , Minerales , Espectroscopía Infrarroja por Transformada de Fourier , TemperaturaRESUMEN
The nickel (Ni) hyperaccumulator Alyssum murale has been developed as a commercial crop for phytoremediation/phytomining Ni from metal-enriched soils. Here, metal co-tolerance, accumulation and localization were investigated for A. murale exposed to metal co-contaminants. A. murale was irrigated with Ni-enriched nutrient solutions containing basal or elevated concentrations of cobalt (Co) or zinc (Zn). Metal localization and elemental associations were investigated in situ with synchrotron X-ray microfluorescence (SXRF) and computed-microtomography (CMT). A. murale hyperaccumulated Ni and Co (> 1000 microg g(-1) dry weight) from mixed-metal systems. Zinc was not hyperaccumulated. Elevated Co or Zn concentrations did not alter Ni accumulation or localization. SXRF images showed uniform Ni distribution in leaves and preferential localization of Co near leaf tips/margins. CMT images revealed that leaf epidermal tissue was enriched with Ni but devoid of Co, that Co was localized in the apoplasm of leaf ground tissue and that Co was sequestered on leaf surfaces near the tips/margins. Cobalt-rich mineral precipitate(s) form on leaves of Co-treated A. murale. Specialized biochemical processes linked with Ni (hyper)tolerance in A. murale do not confer (hyper)tolerance to Co. A. murale relies on a different metal storage mechanism for Co (exocellular sequestration) than for Ni (vacuolar sequestration).
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Brassicaceae/metabolismo , Cobalto/metabolismo , Metales/metabolismo , Níquel/metabolismo , Biodegradación Ambiental , Calcio/química , Calcio/metabolismo , Cobalto/química , Manganeso/química , Manganeso/metabolismo , Metales/química , Níquel/química , Hojas de la Planta/metabolismo , Suelo/análisis , Zinc/química , Zinc/metabolismoRESUMEN
The publisher regrets that this was an accidental duplication of an article that has already been published in Eur. J. Echocardiogr., 4 (2003) 162-168, . The duplicate article has therefore been withdrawn.
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AIMS: The scope of this study was to assess the potential value of pulmonary venous flow diastolic deceleration time to predict end-diastolic pressure and stratify patients with regard to elevation of left ventricular end-diastolic pressures. METHODS AND RESULTS: In 174 consecutive patients, pulmonary venous flow diastolic deceleration time was determined and compared with left ventricular end-diastolic pressures measured invasively. The sample was randomly divided into two subgroups of equal size for modelling of prediction and independent testing of the model. Predicted left ventricular end-diastolic pressures calculated from pulmonary venous flow diastolic deceleration time (left ventricular end-diastolic pressures=-10.87 + 5261/pulmonary venous flow diastolic deceleration time) agreed well with measured left ventricular end-diastolic pressures (mean difference: -1.3 +/- 3.4 mmHg). The correlation of left ventricular end-diastolic pressures with pulmonary venous flow diastolic deceleration time is fair (r=0.73989). A value of pulmonary venous flow diastolic deceleration time <220 ms is suggestive of elevated left ventricular end-diastolic pressures and should be monitored. A value of pulmonary venous flow diastolic deceleration time <190 ms predicts elevated left ventricular end-diastolic pressures. A value of pulmonary venous flow diastolic deceleration time <165 ms predicts severely elevated left ventricular end-diastolic pressures. With 190 ms as a cut-off value for elevated and 165 ms for severely elevated left ventricular end-diastolic pressures, cross-table analysis classifies all patients with normal left ventricular end-diastolic pressures correctly. No patient with severe elevation (<18 mmHg) of left ventricular end-diastolic pressures is classified as normal (chi2=102, P<0.0001). CONCLUSION: Pulmonary venous flow diastolic deceleration time is an appropriate non-invasive measurement to stratify patients with respect to elevation of left ventricular end-diastolic pressures.
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Desaceleración , Venas Pulmonares/fisiopatología , Volumen Sistólico/fisiología , Presión Ventricular/fisiología , Adulto , Anciano , Anciano de 80 o más Años , Cardiomiopatía Dilatada/diagnóstico , Cardiomiopatía Dilatada/fisiopatología , Enfermedad de la Arteria Coronaria/diagnóstico , Enfermedad de la Arteria Coronaria/fisiopatología , Ecocardiografía , Femenino , Alemania , Ventrículos Cardíacos/diagnóstico por imagen , Ventrículos Cardíacos/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Insuficiencia de la Válvula Mitral/diagnóstico , Insuficiencia de la Válvula Mitral/fisiopatología , Valor Predictivo de las Pruebas , Venas Pulmonares/diagnóstico por imagen , Índice de Severidad de la Enfermedad , Estadística como Asunto , Factores de TiempoRESUMEN
The contribution of the angiotensin (Ang) II type 2 receptor (AT2R) to cardiac hypertrophy is still controversial. Here we examined the effect of overexpressing the human AT2R in cultured porcine cardiac fibroblasts (pFib) on proliferation, procollagen I mRNA expression, and - as putatively underlying signal-transduction pathways - on mitogen-activated protein kinase ERK1/ERK2 and phosphotyrosine phosphatase activities. As quantitated by 125I-(Sar1,Ile8)-Ang II binding, transduction of cardiac fibroblasts with the adenoviral AT2R expression vector led to a six- to tenfold higher AT2 than endogenous Ang II type 1 receptor (AT1R) expression. The overexpressed AT2R had the same apparent molecular mass as the endogenous AT2R in rat PC12W cells. Proliferation was not significantly lower in AT2R expressing pFib than in antisense-transduced controls (TA2) upon stimulation with Ang II (AT2R 110.5+/-4.8% vs. TA2 110.2+/-5.5%), Ang II plus the AT1R blocker Irbesartan (97.1+/-1.4% vs. 108.0+/-5.0; P=0.052) and the partial AT2R antagonist CGP42112 at the agonistic concentration of 50 nM (92.1+/-2.7% vs. 99.8+/-3.1%; P=0.053). Procollagen Ialpha2 (COL1A2) mRNA levels were quantitated by (a) northern blot analysis and (b) reverse transcriptase polymerase chain reaction. COL1A2/GAPDH (a) and COL1A2/beta-actin (b) ratios revealed no differences between AT2R-transduced fibroblasts and antisense controls when stimulated with Ang II (1 microM, 24 h) plus Irbesartan and 10 ng/ml transforming growth factor beta1. Ang II stimulation of the endogenous AT1R increased extracellular signal regulated kinase 1/2 activities. This response was reduced by Irbesartan, but PD123319 had no effect. Time course and magnitude of Ang II stimulated ERK1/ERK2 activation was identical in AT2R-transduced and control cells. Also, neither simultaneous nor Ang II pre-stimulation, suggested to induce gene expression of the MAP kinase phosphatase 1, modulated phorbol myristate acetate-stimulated ERK1/ERK2 activation in AT2R-transduced pFib, in AT2R-transduced human umbilical vein endothelial cells, and in PC12W cells. By the use of a tyrosine phosphatase assay we observed an inhibition of phosphotyrosine phosphatase activity by 30.8% (P=0.009, n=5) after 5 min Ang II stimulation of AT2R-expressing pFib. Immunoprecipitation-tyrosine phosphatase assays revealed that inhibition of phosphotyrosine phosphatase 1B, which regulates insulin signaling, contributed to this effect. In conclusion, stimulation of the overexpressed human AT2R in porcine cardiac fibroblasts inhibited tyrosine phosphatase activity but had no significant effect on fibroblast functions related to cardiac fibrosis. It is conceivable that possible antifibrotic AT2R effects are species specific and/or require the interaction between fibroblasts and cardiomyocytes, probably via paracrine factors, or mechanical load.
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Adenoviridae/metabolismo , Fibroblastos/metabolismo , Miocardio/metabolismo , Receptores de Angiotensina/metabolismo , Adenoviridae/genética , Animales , Western Blotting , División Celular , Células Cultivadas , Colágeno , Colágeno Tipo I/metabolismo , Electroforesis en Gel de Poliacrilamida , Activación Enzimática , Humanos , Sistema de Señalización de MAP Quinasas , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Modelos Genéticos , Fosforilación , Pruebas de Precipitina , Proteína Fosfatasa 1 , Proteína Tirosina Fosfatasa no Receptora Tipo 1 , Proteínas Tirosina Fosfatasas/metabolismo , ARN Mensajero/metabolismo , Receptor de Angiotensina Tipo 2 , Transducción de Señal , Porcinos , Factores de Tiempo , Transducción Genética , Tirosina/metabolismoRESUMEN
Tumor necrosis factor alpha (TNFalpha) interferes with insulin signaling in adipose tissue and may promote insulin resistance. Insulin resistance is associated with vascular injury, but little is known about the interaction of TNFalpha and insulin in the vasculature. By activating the Insulin receptor (IR) --> IRS-1 --> phosphatidylinositol-3-kinase (PI3K) --> Akt-pathway, insulin protects vascular smooth muscle cells (VSMC) from undergoing apoptosis. We therefore investigated the effect of TNFalpha on insulin's antiapoptotic signaling in rat aortic VSMC. Insulin induced rapid tyrosine-phosphorylation of the IR and IRS-1 and caused a 2.8-fold increase of IRS-1-bound PI3K. TNFalpha had no effect on insulin-induced tyrosine-phosphorylation of IR or IRS-1, but inhibited insulin-stimulated IRS-1/PI3K-association by 84%. Insulin-induced phosphorylation of Akt downstream of PI3K was inhibited by TNFalpha in a similar pattern. We next examined the effect of TNFalpha on insulin's protective actions on H(2)O(2)-induced apoptosis. Insulin alone prevented 72.8% of H(2)O(2)-induced apoptosis, which was significantly inhibited by TNFalpha. TNFalpha alone did not induce apoptosis. In contrast, TNFalpha had no effect on PDGF-induced antiapoptotic signal transduction via Akt. Thus, TNFalpha selectively interferes with insulin's antiapoptotic signaling in VSMC by inhibiting the association of IRS-1/PI3K and the downstream activation of Akt.
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Apoptosis , Endotelio Vascular/metabolismo , Insulina/metabolismo , Músculo Liso/citología , Tiazolidinedionas , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Aorta/citología , Western Blotting , Células Cultivadas , Activación Enzimática , Citometría de Flujo , Peróxido de Hidrógeno/farmacología , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Fosforilación , Pruebas de Precipitina , Unión Proteica , Ratas , Ratas Sprague-Dawley , Transducción de Señal , Tiazoles/farmacología , Factores de TiempoRESUMEN
Signal transduction pathways that mediate neuronal commitment to apoptosis involve the nuclear factor kappa B (NF-kappaB) transcription factor. The bcl-x gene is a member of the bcl-2 family of genes that regulate apoptosis, and gives rise to two proteins, Bcl-XL and Bcl-XS, via alternative mRNA splicing. BCl-XL protein, like Bcl-2, is a dominant inhibitor of apoptotic cell death, whereas Bcl-XS promotes apoptosis. While there is high expression of Bcl-XL in the developing and adult brain, few transcriptional control elements have been identified in the bcl-x promoter. There are two functional nuclear factor-kappa B (NF-kappaB) DNA binding sites clustered upstream of the brain-specific transcription start site in the upstream promoter region of murine bcl-x. Recombinant NF-kappaB proteins bind to these sites. Also NF-kappaB overexpression, coupled with bcl-x promoter/reporter assays using a series of murine bcl-x promoter and deletion mutants, has identified the downstream 1.1kb of the bcl-x promoter as necessary for basal promoter activity and induction by NF-kappaB in support of the hypothesis that NF-kappaB can act to enhance BCl-XL expression via highly selective interactions with the bcl-x promoter, where NF-kappaB binding and promoter activation are dependent on specific DNA binding site sequences and NF-kappaB protein dimer composition. Hypoxia induces apoptosis in the hippocampus where the NF-kappaB dimers c-Rel/p50 and p50/pS0 bind to the bcl-x promoter NF-kappaB site.
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Encéfalo/fisiopatología , Hipoxia/genética , Hipoxia/fisiopatología , FN-kappa B/fisiología , Proteínas Proto-Oncogénicas c-bcl-2/genética , Transcripción Genética/fisiología , Animales , Proteína bcl-XRESUMEN
RNA fingerprinting by arbitrarily primed PCR (RAP-PCR) is a powerful tool to screen differential gene expression. However, PCR-based screening techniques show a high incidence of false positive results (40-90%). In order to increase the efficiency and feasibility of RAP-PCR, the original protocol was modified and applied to analyse differential gene expression in human coronary macro- (HCEC) and microvascular (HCMEC) endothelial cells. The major modifications introduced were: (i) the use of two primers for PCR amplification, instead of reverse-transcription primer alone; (ii) the use of three cycles at low stringency followed by further amplification at high stringency; (iii) optimization of amplification cycle number, template amount, and concentration of primers, dNTP, Mg(2+); (iv) detection of fingerprints by silver staining; and (v) direct sequencing using RAP-PCR primers. Analysis of untreated and TNF alpha -stimulated (100 U ml(-1)for 1, 4, and 24 h) HCEC and HCMEC displayed 11 differentially expressed products by 18 primer combinations. Confirmation of results by RT-PCR showed that the rate of false positives attributable to our screening method was less than 20%. Among detected RAP-PCR products, the expression of Mn-superoxide dismutase, A20 zinc finger protein, and three novel genes (A/a, 4/d, 7/c) was more strongly modulated by TNF in HCEC than HCMEC. A further novel gene (B/e) was strongly expressed in HCMEC while only barely detectable in HCEC. In conclusion, modification of RAP-PCR strongly reduced the incidence of false positives, eliminated a radioactive requirement, and allowed sequencing without prior cloning, supplying an improved technology able to identify new differentially expressed genes between macro- and microvascular endothelial cells.
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Vasos Coronarios/metabolismo , Dermatoglifia del ADN/métodos , Endotelio Vascular/metabolismo , Expresión Génica , ARN Mensajero/metabolismo , ARN/química , Células Cultivadas , Cartilla de ADN , Reacciones Falso Positivas , Humanos , Reacción en Cadena de la Polimerasa , ARN/análisis , ARN Mensajero/genética , Técnica del ADN Polimorfo Amplificado Aleatorio , Reproducibilidad de los Resultados , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
BACKGROUND: This study examined whether 34 degrees C or 31 degrees C hypothermia during global cerebral ischemia with hyperglycemic cardiopulmonary bypass (CPB) in surviving pigs improves electroencephalographic (EEG) recovery and histopathologic scores when compared with normothermic animals. METHODS: Anesthetized pigs were placed on CPB and randomly assigned to 37 degrees C (n = 9), 34 degrees C (n = 10), or 31 degrees C (n = 8) management. After increasing serum glucose to 300 mg/dL, animals underwent 15 minutes of global cerebral ischemia by temporarily occluding the innominate and left subclavian arteries. Following reperfusion, rewarming, and termination of CPB, animals were recovered for 24 (37 degrees C animals) or 72 hours (34 degrees C and 31 degrees C animals). Daily EEG signals were recorded, and brain histopathology from cortical, hippocampal, and cerebellar regions was graded by an independent observer. RESULTS: Before ischemia, serum glucose concentrations were similar in the 37 degrees C (307+/-9 mg/dL), 34 degrees C (311+/-14 mg/dL), and 31 degrees C (310+/-15) groups. By the first postoperative day, EEG scores in 31 degrees C animals (4.2+/-0.6) had returned to baseline and were greater than those in the 34 degrees C (3.4+/-0.5) and 37 degrees C (2.5+/-0.4) groups (p < 0.05, respectively, between groups). Cooling to 34 degrees C showed selective improvement over 37 degrees C in hippocampal, temporal cortical, and cerebellar regions, but the greatest improvement in all regions occurred with 31 degrees C. Cumulative neuropathology scores in 31 degrees C animals (13.5+/-2.2) exceeded 34 degrees C (6.8+/-2.2) and 37 degrees C (1.9+/-2.1) animals (p < 0.05, respectively, between groups). CONCLUSIONS: Hypothermia during CPB significantly reduced the morphologic consequences of severe, temporary cerebral ischemia under hyperglycemic conditions, with the greatest protection at 31 degrees C.
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Isquemia Encefálica/patología , Encéfalo/patología , Puente Cardiopulmonar/métodos , Hiperglucemia/complicaciones , Hipotermia Inducida/métodos , Animales , Isquemia Encefálica/etiología , Modelos Animales de Enfermedad , Electroencefalografía/métodos , Femenino , Hemodinámica/fisiología , Miocardio/patología , Probabilidad , Distribución Aleatoria , Proteínas S100/análisis , Sensibilidad y Especificidad , Tasa de Supervivencia , PorcinosRESUMEN
Cell death often occurs after hypoxic/ischemic injury to the central nervous system. Changes in levels of the anti-apoptotic Bcl-X(L) protein may be a determining factor in hypoxia-induced neuronal apoptosis. The transcription factor NF-kappa B regulates bcl-x gene expression. In this study, we examined the role of NF-kappa B in the regulation of bcl-x in hypoxia-induced cell death. Rat hippocampus and basal forebrain tissues were collected at different time points after hypoxia (7%O(2), 93% N(2) for 10 or 20 min). We found that 1) hypoxia induced apoptosis in the hippocampus and basal forebrain; 2) the NF-kappa B dimers c-Rel/p50 and p50/p50 bound to the bcl-x promoter NF-kappa B sequence (CS4) in the hippocampus, but only p50/p50 bound to the CS4 sequence in the basal forebrain and hypoxia-induced differential binding patterns of c-Rel/p50 and p50/p50 correlated with the bcl-x expression pattern in the hippocampus; 3) the hypoxia-induced patterns of binding of c-Rel/p50 to the bcl-x promoter CS4 sequence were different from those to the IgG-kappa B enhancer sequence, whereas those of p50/p50 were similar to both sequences; 4) nuclear protein levels of c-Rel, but not p50, correlated with the c-Rel/p50 DNA binding patterns to the bcl-x CS4 site; and 5) there were differential responses to hypoxia among the different NF-kappa B protein subunits. These results suggest that there is a tissue-specific regulation of bcl-x gene expression by NF-kappa B in hypoxia-induced cell death in the hippocampus. The absence of these regulating features in the basal forebrain may account for the early appearance of apoptosis in response to hypoxia as compared with that in hippocampus.
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Hipocampo/metabolismo , Hipoxia Encefálica/metabolismo , FN-kappa B/metabolismo , Prosencéfalo/metabolismo , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Animales , Apoptosis/fisiología , Masculino , Regiones Promotoras Genéticas/fisiología , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Transcripción Genética/fisiología , Proteína bcl-XRESUMEN
The leukocyte adhesion molecule L-selectin, which mediates the initial steps of leukocyte attachment to vascular endothelium, is intensely glycosylated. Different glycoforms of L-selectin are expressed on different leukocyte subsets and differences in L-selectin glycosylation appear to be correlated with the leukocyte's ability to attach to different endothelial targets. In the present study we addressed the question whether glycosylation of L-selectin influences L-selectin-ligand interactions. To obtain different glycoforms of L-selectin, recombinant proteins were expressed both in the baby hamster kidney (BHK) cell line and in the human myelogenous cell line K562, resulting in sL-sel[BHK] or sL-sel[K562], respectively. The glycosylation characteristics of the purified proteins were determined. The most striking differences in glycosylation were seen in the terminal sialylation. Each of the two proteins carried sialic acids in the alpha 2-3 position, while alpha 2-6-bound sialic acids were found exclusively on sL-sel[K562]. To investigate their adhesive properties, both recombinant sL-selectins were used in cell adhesion assays and interactions with the ligands present on various hematopoietic cell lines or activated human cardiac microvascular endothelial cells were examined. The binding capacity of sL-sel[K562] was about 1.6 fold higher compared to sL-sel[BHK] under static as well as under flow conditions. These findings indicate that the terminal sialylation pattern of L-selectin modulates its binding characteristics.
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Adhesivos/química , Endotelio Vascular/química , Células Madre Hematopoyéticas/química , Selectina L/química , Amidohidrolasas , Adhesión Celular , Línea Celular , Citometría de Flujo , Glicosilación , Humanos , Selectina L/biosíntesis , Selectina L/aislamiento & purificación , Ligandos , Péptido-N4-(N-acetil-beta-glucosaminil) Asparagina Amidasa , Unión Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/aislamiento & purificación , Solubilidad , TransfecciónRESUMEN
This study explored the modulatory effects of nitric oxide and thromboxane A2 on contractions to ergonovine and methylergonovine in human coronary arteries. To elucidate the different role of nitric oxide synthase in the response to the ergot alkaloids, the serotonin (5-HT) receptors involved in nitric oxide synthase in the response to the ergot alkaloids, the 5-HT receptors involved in nitric oxide release and the contraction of the vascular smooth muscle were characterized with more selective 5-HT-receptor agonists and antagonists. Rings of human coronary arteries from explanted hearts were suspended in organ chambers for isometric tension recording. After testing for contractile (potassium chloride, 60 mM) and endothelial function (substance P, 10(-8) M), respectively, they were exposed to ergot alkaloids or other agonists in the absence or presence of U 46619 (10(-9) M), or nitro-L-arginine (10(-4) M), or both. Ergonovine and methylergonovine were comparable, weak vasoconstrictors in untreated preparations. Contractions to ergonovine were augmented by U 46619, but not by nitro-L-arginine. Contractions to methylergonovine were augmented only by combining U 46619 and nitro-L-arginine. Serotonin and methylergonovine, but not ergonovine, elicited endothelium-dependent, nitric oxide-mediated relaxations. Nonselective 5-HT(1B/1D)-receptor stimulation caused both contractions and relaxations; selective 5-HT1B stimulation caused relaxations only. In the human coronary artery, contractions to ergonovine are not dependent on NO release but are synergistically augmented by thromboxane. Methylergonovine causes similar effects on the vascular smooth muscle, but contractions are inhibited by the release of NO from the endothelium. The 5-HT receptor on the endothelium appears to be different from the receptor on the vascular smooth muscle, which mediates the contractile response to the ergot alkaloids.
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Ergonovina/farmacología , Metilergonovina/farmacología , Músculo Liso Vascular/efectos de los fármacos , Óxido Nítrico/farmacología , Oxitócicos/farmacología , Tromboxano A2/farmacología , Vasoconstricción/efectos de los fármacos , Adulto , Niño , Vasos Coronarios/efectos de los fármacos , Interacciones Farmacológicas , Ergonovina/antagonistas & inhibidores , Femenino , Humanos , Masculino , Metilergonovina/antagonistas & inhibidores , Persona de Mediana Edad , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacologíaRESUMEN
The tyrosine kinase receptor Tie2 (also known as Tek) plays an important role in the development of the embryonic vasculature and persists in adult endothelial cells (ECs). Tie2 was shown to be upregulated in tumors and skin wounds, and its ligands angiopoietin-1 and -2, although they are not directly mitogenic, modulate neovascularization. To gain further insight into the regulation of Tie2, we have studied the effect of hypoxia and inflammatory cytokines, two conditions frequently associated with neoangiogenic processes, on Tie2 expression in human ECs. Exposure to 1% O(2) led to a time-dependent significant rise of Tie2 protein levels in human coronary microvascular endothelial cells (HCMECs) and dermal microvascular ECs (HMEC-1) (3.2- and 2.5-fold within 24 hours), which was reversible after reoxygenation, and induced a less marked increase in human umbilical vein ECs (HUVECs; 1.7-fold). Hypoxia-conditioned medium and D-deoxyglucose did not change Tie2 expression, but desferrioxamine and cobalt, which are known to mimic hypoxia-sensing mechanisms, induced Tie2 at ambient oxygen tensions. Tumor necrosis factor-alpha induced Tie2 in a time- and dose-dependent fashion in all 3 EC types (HUVEC, 2.3-fold; HMEC-1, 2. 8-fold; and HCMEC, 3.0-fold; 10 ng/mL, 24 hours). Enhanced expression was also found after exposure to interleukin-1beta (1 ng/mL). Changes in Tie2 protein levels were paralleled by changes in mRNA expression. In accordance with these in vitro findings, immunohistochemistry revealed focal upregulation of Tie2 in capillaries at the border of infarcted human and rat myocardium. In conclusion, the data show that hypoxia and inflammatory cytokines upregulate Tie2, which may contribute to the angiogenic response in ischemic tissues.
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Hipoxia de la Célula , Citocinas/farmacología , Endotelio Vascular/metabolismo , Proteínas Tirosina Quinasas Receptoras/biosíntesis , Animales , Bovinos , Células Cultivadas , Vasos Coronarios , Modelos Animales de Enfermedad , Endotelio Vascular/efectos de los fármacos , Humanos , Inmunohistoquímica , Interleucina-1/farmacología , Infarto del Miocardio/metabolismo , Neovascularización Fisiológica , ARN Mensajero/análisis , Ratas , Proteínas Tirosina Quinasas Receptoras/genética , Receptor TIE-2 , Piel/irrigación sanguínea , Factor de Necrosis Tumoral alfa/farmacología , Venas Umbilicales , Regulación hacia ArribaRESUMEN
A total of 45 pregnant ewes were assigned to one of three treatment groups: 1 g ethanol/kg maternal body weight (n = 18); pair-fed control (n = 15); and ad lib control (n = 12). Dosing started at gestational day (GD) 106, and was administered every other day until GD 134. Parturition occurred between GD 144 and 147. Analysis of the placentas indicated that ethanol exposure decreased cotyledon diameter and cotyledon weight compared to the control groups (p < 0. 05). At birth, lambs were given a Vigor Score and then behavior was assessed using a videotape monitoring system for 24 h. Offspring in the ethanol treatment group were significantly less vigorous at birth (p < 0.05). This finding reversed during the subsequent 24 h such that the ethanol-exposed lambs were significantly more active (p = 0.001) than the control lambs. Morphometric and histologic examination of the cerebral cortex and hippocampus revealed no differences amongst the three treatment groups. Collectively, the data demonstrate that moderate ethanol exposure during the third-trimester equivalent of gestation can produce placental dysmorphology and postnatal behavioral anomalies in neonatal lambs in the absence of gross neurologic injury.
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Alcoholismo , Encéfalo/anatomía & histología , Trastornos Mentales/etiología , Actividad Motora , Efectos Tardíos de la Exposición Prenatal , Aborto Espontáneo , Animales , Animales Recién Nacidos , Encéfalo/citología , Corteza Cerebral/anatomía & histología , Corteza Cerebral/citología , Femenino , Muerte Fetal , Edad Gestacional , Hipocampo/anatomía & histología , Hipocampo/citología , Tamaño de la Camada , Tamaño de los Órganos , Embarazo , Complicaciones del Embarazo , Resultado del Embarazo , Ovinos , Grabación de Cinta de Video , Aumento de PesoRESUMEN
We retrospectively studied 17 cases of distal pectoralis major muscle rupture to compare the results of repair in acute and chronic injuries and to compare operative and nonoperative treatment. Thirteen patients underwent surgery (six acute injuries [less than 2 weeks after injury] and seven chronic injuries) and four had nonoperative management. The mean age of the patients at injury was 29, and 10 of the 17 injuries were the result of weight lifting. Follow-up ranged from 18 months to 6 years (mean, 28 months). All patients subjectively rated strength, pain, motion, function with strenuous sporting activities, cosmesis, and overall satisfaction. Objectively, patients were examined for range of motion, deformity, atrophy, and strength. Isokinetic strength testing was performed in eight patients: six treated operatively (three acute and three chronic) and two treated nonoperatively. Overall subjective ratings were 96% in the acute group, 93% in the chronic group, and only 51% in the nonoperative group. Isokinetic testing showed that patients operated on for acute injuries had the highest adduction strength (102% of the opposite side) compared with patients with chronic injuries (94%) or nonoperative treatment (71%). There were no statistically significant subjective or objective differences in outcome between the patients treated operatively for acute or chronic injuries, but these patients fared significantly better than patients treated nonoperatively.
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Traumatismos en Atletas/cirugía , Músculo Esquelético/cirugía , Levantamiento de Peso/lesiones , Adulto , Traumatismos en Atletas/patología , Femenino , Humanos , Masculino , Músculo Esquelético/patología , Dolor , Rango del Movimiento Articular , Estudios Retrospectivos , Rotura , Tórax/patología , Resultado del TratamientoRESUMEN
While the cellular mechanisms of atherosclerosis have been intensively studied, the mechanisms leading to preferential localization of atherosclerotic lesions are less well understood. To further define these mechanisms, endothelial cells from coronary arteries, i.e., vessels with frequent atherosclerotic lesions, were isolated and grown in vitro. In order to compare the reactions of both cell types, endothelial cells derived from microvessels of human hearts were isolated and cultured under identical conditions. Incubation of endothelial cells with oxidized LDL (75 microg/ml protein) induced a significant increase in PAI-1 activity (182%, p < 0.05) in coronary macrovascular endothelial cells. This stimulatory effect of ox-LDL was less significant in microvascular endothelial cells (144%, p < 0.05). n-LDL did not influence secreted PAI-1 activity. Stimulation with angiotensin II induced expression of E-selectin more effectively in coronary macrovascular than in microvascular endothelial cells. In addition, angiotensin II-induced E-selectin expression led to increased E-selectin-dependent adhesion of HL60 cells to coronary macrovascular endothelial cells under flow conditions, while only little effect was observed with cardiac microvascular endothelial cells. In contrast, L-selectin-dependent adhesion, which has been shown to play an important role in inflammatory reactions, was preferentially observed in cardiac microvascular endothelial cells and could only be stimulated with TNFalpha, not by angiotensin II. Therefore, these cellular differences may in part explain specific properties of cardiac endothelial cells: Such that atherosclerotic lesions are localized in macrovascular vessel segments, whereas inflammatory responses are predominantly found in the microvasculature.