Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 30
Filtrar
1.
J Phys Condens Matter ; 36(37)2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38843803

RESUMO

In this work we examine synthetic antiferromagnetic structures consisting of two, three, and four antiferromagnetic coupled layers, i.e. bilayers, trilayers, and tetralayers. We vary the thickness of the ferromagnetic layers across all structures and, using a macrospin formalism, find that the nearest neighbor exchange interaction between layers is consistent across all structures for a given thickness of the ferromagnetic layer. Our model and experimental results demonstrate significant differences in how the static equilibrium states of even and odd-layered structures evolve as a function of the external field. Even layered structures continuously evolve from a collinear antiferromagnetic state to a spin canted non-collinear magnetic configuration that is mirror-symmetric about the external field. In contrast, odd-layered structures begin with a ferrimagnetic ground state; at a critical field, the ferrimagnetic ground state evolves into a non-collinear state with broken symmetry. Specifically, the magnetic moments found in the odd-layered samples possess stable static equilibrium states that are no longer mirror-symmetric about the external field after a critical field is reached.

2.
J Proteomics ; 149: 15-22, 2016 10 21.
Artigo em Inglês | MEDLINE | ID: mdl-26975722

RESUMO

Integral membrane proteins are generally under-represented in routine proteomic analyses, mostly because of their relatively low abundance, hydrophobicity and lack of trypsin-cleavage sites. To increase the coverage of membrane proteomes, various strategies have been developed, targeting mostly the extra-membrane segments of membrane proteins. We focused our attention to the rather overlooked hydrophobic transmembrane segments. Such peptides can be isolated after carbonate stripping and protease "shaving" of membranes isolated by simple centrifugation procedure. The treated membranes with embedded hydrophobic peptides can then be solubilized in organic solvents, re-digested with CNBr, delipidated and subjected to LC-MS/MS analysis. We modified the original "hppK" method, and applied it for the analysis of human lymphoma cells. We identified 1224 proteins of which two-thirds were IMPs with 1-16 transmembrane segments. This method allowed us to identify 13 "missing proteins" - proteins with no previous evidence on protein level. BIOLOGICAL SIGNIFICANCE: Integral membrane proteins execute numerous essential functions and represent substantial part of eukaryotic proteomes. Our knowledge of their function and expression is, however, limited. Novel approaches extending our knowledge of membrane proteome are therefore highly desired. As we demonstrate here, a non-conventional method which targets rather overlooked hydrophobic transmembrane segments of integral membrane proteins has wide potential to provide the missing information on the membrane proteome. We show that it can deliver identification and potentially also quantification of hundreds of integral membrane proteins including the so called "missing proteins".


Assuntos
Linfoma de Célula do Manto/química , Proteínas de Membrana/análise , Proteínas de Neoplasias/análise , Proteoma/química , Proteômica/métodos , Tripsina/química , Cromatografia Líquida/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Peptídeos/análise , Espectrometria de Massas em Tandem/métodos
3.
Phys Rev Lett ; 116(5): 057601, 2016 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-26894733

RESUMO

Because of its transverse nature, spin Hall effects (SHE) provide the possibility to excite and detect spin currents and magnetization dynamics even in magnetic insulators. Magnetic insulators are outstanding materials for the investigation of nonlinear phenomena and for novel low power spintronics applications because of their extremely low Gilbert damping. Here, we report on the direct imaging of electrically driven spin-torque ferromagnetic resonance (ST-FMR) in the ferrimagnetic insulator Y_{3}Fe_{5}O_{12} based on the excitation and detection by SHEs. The driven spin dynamics in Y_{3}Fe_{5}O_{12} is directly imaged by spatially resolved microfocused Brillouin light scattering spectroscopy. Previously, ST-FMR experiments assumed a uniform precession across the sample, which is not valid in our measurements. A strong spin-wave localization in the center of the sample is observed indicating the formation of a nonlinear, self-localized spin-wave "bullet".

4.
Phys Rev Lett ; 111(7): 077201, 2013 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-23992078

RESUMO

We have patterned novel Permalloy thin films with quasicrystalline Penrose P2 tilings and measured their dc magnetization and ferromagnetic resonance absorption. Reproducible anomalies in the hysteretic, low-field data signal a series of abrupt transitions between ordered magnetization textures, culminating in a smooth evolution into a saturated state. Micromagnetic simulations compare well to experimental dc hysteresis loops and ferromagnetic resonance spectra and indicate that systematic control of magnetic reversal and domain wall motion can be achieved via tiling design, offering a new paradigm of magnonic quasicrystals.

5.
Clin Exp Allergy ; 41(7): 1031-43, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21623965

RESUMO

BACKGROUND: The incidence of food allergy to wheat is increasing. Its diagnosis depends on the purity of major allergens and their inclusion in tests. Isolation and characterization of wheat allergens are therefore of utmost importance. OBJECTIVE: To purify and identify wheat flour allergens most frequently recognized by patients' IgE antibodies and to study their allergenicity. METHODS: Water/salt-soluble extracts from wheat flour were prepared and separated using a combination of ultrafiltration, isoelectric focusing and liquid chromatography. Purified proteins were analysed by immunoblotting using pooled sera from patients with atopic dermatitis who possessed IgE specific to wheat. Wheat proteins found to bind IgE were subsequently identified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. The frequency and intensity of IgE binding of isolated proteins were tested using individual sera from patients and controls. RESULTS: We developed a procedure that allows isolation of wheat allergens from natural sources. Twenty-seven potential wheat allergens have been successfully identified; of these, the following seven are newly reported in food allergy: endogenous α-amylase/subtilisin inhibitor, trypsin/α-amylase inhibitor (AAI) CMX1/CMX3, thaumatin-like protein (TLP), xylanase inhibitor protein-1, ß-glucosidase, class II chitinase and 26 kDa endochitinase. TLP and wheatwin were shown to activate patients' basophils to a similar extent as two well-known allergens, lipid transfer protein (Tri a 14) and AAI 0.19 (Tri a 28.0101). CONCLUSION AND CLINICAL RELEVANCE: Our new approach enables the isolation of water/salt-soluble wheat allergens in their native form in amounts sufficient both for biological testing (in vivo and in vitro) and for physicochemical characterization. Such studies will lead to a more detailed knowledge of allergenicity of wheat proteins and to improved accuracy of diagnostic tests.


Assuntos
Alérgenos/isolamento & purificação , Basófilos/imunologia , Farinha/análise , Imunoglobulina E/sangue , Proteínas de Plantas/isolamento & purificação , Triticum/imunologia , Adolescente , Adulto , Alérgenos/química , Alérgenos/imunologia , Criança , Pré-Escolar , Cromatografia Líquida de Alta Pressão , Dermatite Atópica/diagnóstico , Dermatite Atópica/imunologia , Feminino , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Humanos , Focalização Isoelétrica , Masculino , Pessoa de Meia-Idade , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Triticum/química , Ultrafiltração , Hipersensibilidade a Trigo/diagnóstico , Hipersensibilidade a Trigo/imunologia , Adulto Jovem
6.
Am J Physiol Heart Circ Physiol ; 294(4): H1651-7, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18310524

RESUMO

An exaggerated increase in pulmonary arterial pressure is the hallmark of high-altitude pulmonary edema (HAPE) and is associated with endothelial dysfunction of the pulmonary vasculature. Whether the myocardial circulation is affected as well is not known. The aim of this study was, therefore, to investigate whether myocardial blood flow reserve (MBFr) is altered in mountaineers developing HAPE. Healthy mountaineers taking part in a trial of prophylactic treatment of HAPE were examined at low (490 m) and high altitude (4,559 m). MBFr was derived from low mechanical index contrast echocardiography, performed at rest and during submaximal exercise. Among 24 subjects evaluated for MBFr, 9 were HAPE-susceptible individuals on prophylactic treatment with dexamethasone or tadalafil, 6 were HAPE-susceptible individuals on placebo, and 9 persons without HAPE susceptibility served as controls. At low altitude, MBFr did not differ between groups. At high altitude, MBFr increased significantly in HAPE-susceptible individuals on treatment (from 2.2 +/- 0.8 at low to 2.9 +/- 1.0 at high altitude, P = 0.04) and in control persons (from 1.9 +/- 0.8 to 2.8 +/- 1.0, P = 0.02), but not in HAPE-susceptible individuals on placebo (2.5 +/- 0.3 and 2.0 +/- 1.3 at low and high altitude, respectively, P > 0.1). The response to high altitude was significantly different between the two groups (P = 0.01). There was a significant inverse relation between the increase in the pressure gradient across the tricuspid valve and the change in myocardial blood flow reserve. HAPE-susceptible individuals not taking prophylactic treatment exhibit a reduced MBFr compared with either treated HAPE-susceptible individuals or healthy controls at high altitude.


Assuntos
Doença da Altitude/prevenção & controle , Carbolinas/uso terapêutico , Dexametasona/uso terapêutico , Reserva Fracionada de Fluxo Miocárdico/efeitos dos fármacos , Glucocorticoides/uso terapêutico , Hemodinâmica/efeitos dos fármacos , Inibidores de Fosfodiesterase/uso terapêutico , Edema Pulmonar/prevenção & controle , Aclimatação , Adulto , Altitude , Doença da Altitude/diagnóstico por imagem , Doença da Altitude/fisiopatologia , Método Duplo-Cego , Exercício Físico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Montanhismo , Edema Pulmonar/diagnóstico por imagem , Edema Pulmonar/fisiopatologia , Tadalafila , Fatores de Tempo , Ultrassonografia , Vasodilatação/efeitos dos fármacos
7.
Folia Microbiol (Praha) ; 51(6): 573-8, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17455794

RESUMO

Dye decolorization capacity of two white-rot fungi, Irpex lacteus and Phanerochaete chrysosporium, was compared in N-limited liquid cultures. The agitated cultures showed lower ability to decolorize azo dyes Reactive Orange 16 and Naphthol Blue Black than static cultures. Similar effect was also observed with other structurally different synthetic dyes. The effect of surfactants on the decolorization process is discussed. A significant increase in the Reactive Orange 16 decolorization by the agitated I. lacteus cultures was observed after adding 0.1% Tween 80, following a higher Mn-dependent peroxidase production. The in vitro dye decolorization using the purified enzyme proved its decolorization ability.


Assuntos
Basidiomycota/enzimologia , Corantes/metabolismo , Peroxidases/metabolismo , Compostos Azo/metabolismo , Basidiomycota/efeitos dos fármacos , Basidiomycota/metabolismo , Indução Enzimática , Phanerochaete/enzimologia , Phanerochaete/metabolismo , Polissorbatos/farmacologia
8.
Biochem Soc Trans ; 32(Pt 5): 764-5, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15494009

RESUMO

beta-N-Acetylhexosaminidase from a filamentous fungus Aspergillus oryzae is a secreted enzyme known to be an important component of the binary chitinolytic system. Cloning of the hexA gene and sequencing of the enzyme revealed its unique preproprotein structure. While the enzyme's zincin-like and catalytic domain had significant similarities with members of the glycohydrolase 20 family, the propeptide was unique for the fungal enzyme. Detailed pulse-chase and inhibition studies revealed that propeptide was processed during the biosynthesis of the enzyme. Moreover, the presence of propeptide was necessary for enzyme activation, dimerization and secretion. The catalytic unit was N-glycosylated, and the propeptide was O-glycosylated, both in their C-terminal parts. Deglycosylation experiments revealed that the N-glycosylation increased the stability and solubility of the enzyme. In contrast, O-glycosylated propeptide was necessary to attain the full enzymic activity.


Assuntos
Aspergillus oryzae/enzimologia , Glicosilação , Hexosaminidases/química , Peptídeos/química , Domínio Catalítico , Quitina/química , Cromatografia em Gel , Clonagem Molecular , Hexosaminidase A , Concentração de Íons de Hidrogênio , Estrutura Terciária de Proteína , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Fatores de Tempo , beta-N-Acetil-Hexosaminidases/química
9.
Circulation ; 104(20): 2471-7, 2001 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-11705827

RESUMO

BACKGROUND: We hypothesized that by detecting regions with adequate collateral-derived myocardial blood flow (MBF) within the risk area (RA), we could predict ultimate infarct size (IS) at the time of coronary occlusion. METHODS AND RESULTS: Group 1 dogs (n=15) underwent coronary occlusion without reperfusion, whereas group 2 dogs (n=6) underwent both occlusion and reperfusion. RA was measured with aortic root injections of microbubbles. Myocardial contrast echocardiography (MCE) was performed with high mechanical index intermittent harmonic imaging at pulsing intervals (PIs) of <1 to 30 cardiac cycles during an intravenous infusion of microbubbles (Sonozoid). MBF was measured with radiolabeled microspheres, and postmortem tissue staining was used to determine IS. Perfusion defect size (PDS) on MCE varied with the PI and was largest at a PI of 2.6+/-0.4 seconds, where it correlated well with RA (r=0.82). PDS was smallest at a PI of >/=10.6+/-1.5 seconds, where it correlated closely with IS (r>/=0.92). Areas that underwent necrosis could be identified early after coronary occlusion as having the lowest microvascular flow velocity (beta) and MCE-derived MBF (Axbeta). The results were similar with or without reperfusion. Because of variability in collateral-derived MBF, there was no correlation between RA and ultimate IS (P=0.37). The extent of regional dysfunction also correlated poorly with IS (r=0.31). CONCLUSIONS: MCE can be used immediately after coronary occlusion to define ultimate IS by measuring the magnitude and spatial extent of collateral-derived residual MBF within the RA. Thus, it could help individualize risk and management in acute myocardial infarction.


Assuntos
Circulação Colateral , Circulação Coronária , Doença das Coronárias/diagnóstico por imagem , Ecocardiografia/métodos , Infarto do Miocárdio/diagnóstico por imagem , Animais , Doença das Coronárias/patologia , Doença das Coronárias/fisiopatologia , Cães , Previsões , Infarto do Miocárdio/patologia , Infarto do Miocárdio/fisiopatologia , Reperfusão Miocárdica
10.
Circulation ; 103(21): 2624-30, 2001 May 29.
Artigo em Inglês | MEDLINE | ID: mdl-11382734

RESUMO

BACKGROUND: Power Doppler is a new imaging method for detecting microbubbles during myocardial contrast echocardiography (MCE) based on the registration of variance resulting from ultrasound-induced nonlinear bubble behavior. We tested the hypothesis that power Doppler imaging can be used to quantify coronary stenoses. METHODS AND RESULTS: Three left anterior descending (LAD) coronary stenoses of varying severity were created in each of 9 open-chest dogs. MCE was performed by continuous intravenous infusion of a nitrogen-filled bilayer shell microbubble, PB127, during triggered power Doppler imaging at incremental pulsing intervals. MCE and radiolabeled microsphere measurements were made at baseline and during each stenosis, with and without adenosine stress. Videointensities in the LAD and left circumflex (LCx) beds were plotted against pulsing interval and fit to a previously described exponential function modeling microbubble destruction and replenishment, which was used to derive parameters of bubble velocity (beta) and peak plateau videointensity (A). Contrast defects matching the location of radiolabeled microsphere hypoperfusion were clearly seen, without need for image processing. The product of beta and A was linearly related to LAD/LCx flow (r=0.90, P<0.0001) and inversely related to stenosis gradient (r=-0.70, P<0.0001). Endocardial/epicardial flow ratios were visualized and quantifiable. CONCLUSIONS: As with B-mode harmonics, a model of microbubble destruction/replenishment can be applied to power Doppler data as a means to detect a broad range of stenoses. Image clarity and the lack of attenuation or requirement for background subtraction are additional advantages of this imaging approach. Power Doppler MCE imaging holds promise for the detection of coronary artery disease.


Assuntos
Doença das Coronárias/diagnóstico por imagem , Ultrassonografia Doppler , Animais , Circulação Coronária , Doença das Coronárias/patologia , Vasos Coronários/diagnóstico por imagem , Vasos Coronários/fisiopatologia , Cães , Ecocardiografia
11.
J Am Soc Echocardiogr ; 13(2): 124-30, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10668015

RESUMO

This prospective study was performed to test the hypothesis that the yield of 2-dimensional echocardiography (2DE) would be higher when it is ordered by a cardiologist than by a noncardiologist. Patients referred for transthoracic 2DE for the evaluation of left ventricular systolic function for the 11-month period between July 10, 1995, and June 10, 1996, were included in the study. Demographic, historical, and clinical findings were recorded. Whether the patient was referred by a cardiologist versus a noncardiologist was used as the predictor variable in a binary logistic regression analysis. To address the possibility that the yield of 2DE may be higher for cardiologists because the prevalence of disease in patients referred to them may be higher (selection bias), the analysis was subjected to a propensity score adjustment. Of 2176 patients referred for 2DE during the study, 1033 were referred for the evaluation of left ventricular function. The test had a positive yield in 52% of patients for cardiologists versus 31% for noncardiologists (chi(2) = 45.5, P <.0001, odds ratio 2.4 [CI = 1. 9-3.1]). This difference remained highly significant even when propensity score risk adjustment was made (chi(2) = 54.2, P <.0001, odds ratio 2.0 [CI = 1.5-2.8]). We conclude that the yield of 2DE is higher for cardiologists compared with noncardiologists and that this result was not related to differences in patient populations examined by the two groups. Thus, more efficient use of 2DE may be achieved if patients are referred to cardiologists rather than directly sent for 2DE.


Assuntos
Ecocardiografia/estatística & dados numéricos , Encaminhamento e Consulta , Função Ventricular Esquerda , Idoso , Cardiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Procedimentos Desnecessários
12.
Am Heart J ; 139(3): 394-8, 2000 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-10689252

RESUMO

BACKGROUND: We hypothesized that patients could be selected for echocardiographic evaluation of left ventricular (LV) systolic function on the basis of historic, clinical, radiographic, and electrocardiographic criteria. METHODS AND RESULTS: We prospectively evaluated 300 consecutive inpatients referred for the echocardiographic assessment of LV function, of whom 124 (41%) had LV systolic dysfunction (LVSD) (LV ejection fraction <0.45). Among the historic variables, male sex was the only predictor of LVSD, whereas of the abnormal physical and radiographic findings, cardiomegaly on chest radiography was the only predictor. Among the electrocardiographic findings, the presence of left bundle branch block was positively correlated with the presence of LVSD, whereas a normal electrocardiogram was negatively correlated with this finding. Only 2 patients with LVSD had a normal electrocardiogram. The addition of significant predictors on physical examination and chest radiography doubled the predictive value of the historic variables for determining LVSD. The addition of electrocardiographic findings further doubled the predictive value of the model. Almost 45% of the predictive power of the final multivariate model (chi-square of 48 of the total chi-square of 108) was based on the absence of normal electrocardiogram in patients with LVSD. When chest radiographic findings were excluded from the model, the overall predictive power of the model did not change, with the normal electrocardiogram gaining greater prominence: Full 56% of the predictive power of the model (chi-square of 60 of the total chi-square of 108) resided in the ability of a normal electrocardiogram to discriminate between patients with and those without LVSD. CONCLUSIONS: Historic, chest radiographic, and electrocardiographic variables can be used to predict low likelihood of LVSD on echocardiography. In particular, when the electrocardiogram is normal, it is extremely unlikely to have LVSD. It can be argued that such patients should not be referred for echocardiography.


Assuntos
Ecocardiografia/normas , Seleção de Pacientes , Disfunção Ventricular Esquerda/diagnóstico , Função Ventricular Esquerda/fisiologia , Idoso , Bloqueio de Ramo/diagnóstico , Cardiomegalia/diagnóstico por imagem , Cardiomegalia/epidemiologia , Distribuição de Qui-Quadrado , Técnicas de Apoio para a Decisão , Ecocardiografia/economia , Eletrocardiografia , Feminino , Humanos , Incidência , Funções Verossimilhança , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Exame Físico , Valor Preditivo dos Testes , Estudos Prospectivos , Radiografia , Distribuição por Sexo , Sístole , Disfunção Ventricular Esquerda/epidemiologia
13.
Am Heart J ; 139(2 Pt 1): 231-40, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10650295

RESUMO

BACKGROUND: The aim of this study was to perform a quantitative comparison between myocardial contrast echocardiography (MCE) and single-photon emission computed tomography (SPECT) in patients with prior myocardial infarction (MI). We also wanted to determine the optimal method for the intravenous administration of an ultrasound contrast agent in the clinical setting. METHODS AND RESULTS: Seventeen patients with resting perfusion defects in a single vascular territory on SPECT were studied. MCE was performed with intermittent harmonic imaging during continuous infusions of a second-generation ultrasound contrast agent (Sonovue, Bracco Diagnostics) in all 17 patients and after bolus injection in 8 of them. During continuous infusions, the video intensity (VI) ratio between the abnormal and normal myocardium at a pulsing interval (PI) of 8 cardiac cycles correlated well with the activity ratio between these segments on SPECT (r = 0.73, P <.01). When information regarding microbubble velocity (MV) denoted as change in VI with increasing PIs was added, the correlation with SPECT activity ratio improved (P <.05) significantly (r = 0.87, P <.0001). Higher microbubble doses resulted in higher VI during continuous infusions with good myocardial opacification and no far-field attenuation until the highest dose was reached. With bolus injections, the VI ratio between the abnormal and normal myocardium at PI of 1 and 5 cardiac cycles showed a modest correlation (r = 0.46 and r = 0.48, respectively, P <.05) with activity ratios between these regions on SPECT. When a dose of microbubbles administered as a bolus produced adequate myocardial opacification, it invariably resulted in far-field attenuation. CONCLUSIONS: In patients with prior MI, quantitative assessment of resting perfusion defects on MCE correlates well with regional activity on SPECT. Continuous infusions offer an advantage over bolus injections because they can provide an assessment of both relative VI and MV. Adjustment of the microbubble infusion rate produces adequate myocardial opacification without attenuation.


Assuntos
Circulação Coronária/fisiologia , Vasos Coronários/fisiologia , Coração/fisiologia , Infarto do Miocárdio/fisiopatologia , Tomografia Computadorizada de Emissão de Fóton Único , Vasos Coronários/diagnóstico por imagem , Vasos Coronários/fisiopatologia , Feminino , Hemodinâmica/fisiologia , Humanos , Infusões Intravenosas , Masculino , Microesferas , Infarto do Miocárdio/diagnóstico por imagem , Ultrassonografia
14.
Protein Sci ; 8(7): 1551-6, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10422846

RESUMO

The complete covalent structure of a novel boar DQH sperm surface protein resistant to many classical procedures of enzymatic fragmentation was determined. The relative molecular mass of the major form of this protein determined by ESI-MS and MALDI-MS was 13,065.2+/-1.0 and 13,065.1, respectively. However, additional peaks differing by 162 Da (i.e., minus hexose), 365 Da (i.e., minus hexose and N-acetylhexosamine), 146 Da (i.e., plus deoxyhexose), and 291 Da (i.e., plus sialic acid) indicated the heterogeneity due to differences in glycosylation. The complete covalent structure of the protein was determined using automated Edman degradation, MALDI-MS, and post-source decay (PSD) MALDI-MS, and shown to consist of N-terminal O-glycosylated peptide followed by two fibronectin type II repeats. The carbohydrates are O-glycosidically linked to threonine 10, as confirmed by PSD MALDI-MS of the isolated N-terminal glycopeptide. Eight cysteine residues of the protein form four disulfide bridges, the positions of which were assigned from MALDI-MS and Edman degradation data. We conclude that mass spectral techniques provide an indispensable tool for the detailed analysis of the covalent structure of proteins, especially those that are refractory to standard approaches of protein chemistry.


Assuntos
Proteínas de Transporte/química , Glicoproteínas/química , Glicoproteínas de Membrana/química , Sequência de Aminoácidos , Animais , Glicosilação , Masculino , Dados de Sequência Molecular , Peso Molecular , Conformação Proteica , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Suínos
15.
J Am Soc Echocardiogr ; 12(2): 85-93, 1999 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9950966

RESUMO

Optimal assessment of myocardial perfusion with contrast echocardiography by using B-mode imaging often requires image alignment and background subtraction, which are time consuming and need extensive expertise. Flash echocardiography is a new technique in which primary images are gated to the electrocardiogram and secondary images are obtained by transmitting ultrasound pulses in rapid succession after each primary image. Myocardial opacification is seen in the primary image and not in the secondary images because of ultrasound-induced bubble destruction. Because the interval between the primary and first few secondary images is very short, cardiac motion between these images should be minimal. Therefore we hypothesized that 1 or more secondary images could be subtracted from the primary image without the need for image alignment. The ability of ultrasound to destroy microbubbles was assessed by varying the sampling rate, line density, and mechanical index in 6 open-chest dogs. The degree of translation between images was quantified in the x and y directions with the use of computer cross-correlation. At sampling rates of 158 Hz or less and a mechanical index of more than 0.6, videointensity rapidly declined to baseline levels by 25 ms. Significant translation between images was noted only at intervals of more than 112 ms. It is concluded that flash echocardiography can be used for digital subtraction of baseline from contrast-enhanced B-mode images without image alignment. Background subtraction is therefore feasible on-line, potentially eliminating the need for off-line image processing in the future.


Assuntos
Meios de Contraste , Ecocardiografia/métodos , Técnica de Subtração , Animais , Cães , Eletrocardiografia , Processamento de Imagem Assistida por Computador
16.
Circulation ; 98(18): 1912-20, 1998 Nov 03.
Artigo em Inglês | MEDLINE | ID: mdl-9799213

RESUMO

BACKGROUND: We hypothesized that by using our newly defined method of destroying microbubbles and measuring their rate of tissue replenishment, we could assess the transmural distribution of myocardial perfusion. METHODS AND RESULTS: We studied 12 dogs before and after creation of left anterior descending coronary artery stenoses both at rest and during hyperemia (n=62 stages). Microbubbles were administered as a constant infusion, and myocardial contrast echocardiography (MCE) was performed with the use of different pulsing intervals. The video intensity versus pulsing interval plots derived from each myocardial pixel were fitted to an exponential function: y=A(1-ebetat), where A reflects microvascular cross-sectional area (or myocardial blood volume), and beta reflects mean myocardial microbubble velocity. The product A . beta represents myocardial blood flow (MBF). Average values for these parameters were derived from the endocardial and epicardial regions of interest placed over the left anterior descending coronary artery bed. Radiolabeled microsphere-derived MBF was also measured from the same regions. There was poor correlation between radiolabeled microsphere-derived MBF and A-endocardial/epicardial ratios (EER) (r=0.46). The correlation with beta-EER was better (r=0. 69, P<0.01). The best correlation with radiolabeled microsphere-derived MBF-EER was noted with A . beta-EER (r=0.88, P<0. 01). CONCLUSIONS: The transmural distribution of myocardial perfusion can be accurately assessed with MCE with the use of our newly described method of tissue replenishment of microbubbles after their ultrasound-induced destruction. In the model studied, an uncoupling of the transmural distribution of MBF and myocardial blood volume was observed during reversal of the MBF-EER.


Assuntos
Circulação Coronária/fisiologia , Ecocardiografia , Ar , Animais , Volume Sanguíneo/fisiologia , Meios de Contraste , Doença das Coronárias/diagnóstico por imagem , Doença das Coronárias/fisiopatologia , Cães , Endocárdio , Processamento de Imagem Assistida por Computador , Microesferas , Pericárdio
17.
Biochem Biophys Res Commun ; 238(1): 149-53, 1997 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-9299469

RESUMO

NKR-P1 represent a family of activating receptors in rodent natural killer cells related to C-type animal lectins. We identify here the elements involved in the reactivity of the major receptor of rat, NKR-P1A, with N-linked oligosaccharides of glycoproteins. Plate inhibition assays with isolated, structurally defined N-glycans as inhibitors of binding of NKR-P1A to GlcNAc16-BSA revealed that the removal of both the external sialic acids and the penultimate galactose residues resulted in attaining of significant inhibitory activities. Surprisingly, additional plate inhibition and glycoprotein overlay experiments brought evidence that the core chitobiose, depending on its substitution, can per se support the interaction with NKR-P1A. In a series of linear chitooligomers (n = 2-7), the inhibitory activities reached a maximum for the chitotetraose. The ability of NKR-P1 to recognize both the periphery and the core region of complex type oligosaccharides may define its dual specificity towards carbohydrate components of eukaryotic (e.g., tumor) cell surfaces, but also reflect an evolutionarily conserved reactivity with microbial saccharides important in immune recognition and signaling functions.


Assuntos
Antígenos de Superfície/metabolismo , Dissacarídeos/metabolismo , Células Matadoras Naturais/metabolismo , Lectinas Tipo C , Polissacarídeos/metabolismo , Receptores Imunológicos/metabolismo , Animais , Configuração de Carboidratos , Dissacarídeos/química , Glicosilação , Ácido N-Acetilneuramínico/química , Ácido N-Acetilneuramínico/metabolismo , Subfamília B de Receptores Semelhantes a Lectina de Células NK , Oligossacarídeos/metabolismo , Ovalbumina/metabolismo , Ovomucina/metabolismo , Polissacarídeos/química , Ratos
18.
Circulation ; 94(6): 1430-40, 1996 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-8823003

RESUMO

BACKGROUND: We hypothesized that microvascular reserve is a better indicator of the extent of viable myocardium postinfarction than contractile reserve, especially in the presence of a residual stenosis of the infarct-related artery. METHODS AND RESULTS: Fifteen dogs with various infarct sizes were studied after reperfusion. Contractile reserve, studied by use of dobutamine echocardiography, and microvascular reserve, studied by use of myocardial contrast echocardiography, were measured both before and after creation of a stenosis. In the absence of a stenosis, the relation between infarct size, expressed as percent of risk area, and wall thickening improved with increasing doses of dobutamine (r = .41, .71, and .90 for 5, 10, and 15 micrograms.kg-1.min-1, respectively; P < .01 for dobutamine 15 micrograms.kg-1.min-1). In the presence of a stenosis, however, the relation was poor for all doses of dobutamine (r = .22, .57, and .32 for 5, 10, and 15 micrograms.kg-1.min-1, respectively; P < .01 for 15 micrograms.kg-1.min-1 dobutamine in the absence of a stenosis). There was a fair correlation between infarct size and perfusion defect size on myocardial contrast echocardiography after reperfusion (r = .82), with the defect size underestimating infarct size by approximately 20%. This relationship improved (P < .01) during infusions of both adenosine (r = .99) and dobutamine (r = .94) in the absence of a stenosis. The correlations between infarct size and perfusion defect on myocardial contrast echocardiography also remained good in the presence of a stenosis (r = .95 and .81 for adenosine and dobutamine, respectively; P = NS compared with stenosis). CONCLUSIONS: Microvascular reserve is superior to contractile reserve for definition of the spatial topography of necrosis and hence the extent of viable myocardium within the infarct bed after reperfusion, particularly when a residual stenosis is present in the infarct-related artery.


Assuntos
Circulação Coronária , Doença das Coronárias/fisiopatologia , Contração Miocárdica , Isquemia Miocárdica/fisiopatologia , Reperfusão Miocárdica , Terapia de Salvação , Adenosina/farmacologia , Animais , Cardiotônicos/farmacologia , Fármacos Cardiovasculares/farmacologia , Meios de Contraste , Dobutamina/farmacologia , Cães , Ecocardiografia , Microcirculação , Isquemia Miocárdica/diagnóstico por imagem
19.
Circulation ; 94(4): 748-54, 1996 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-8772698

RESUMO

BACKGROUND: The aim of this study was to determine whether myocardial contrast echocardiography (MCE) during exogenous vasodilation can accurately delineate infarct size, and hence the extent of myocardial viability, both immediately (15 minutes) and late (3 hours) after reperfusion when postreflow coronary hyperema is still present. METHODS AND RESULTS: Twenty-one open-chest anesthetized dogs underwent 3 to 6 hours of coronary occlusion followed by reperfusion. MCE was performed 15 minutes after reflow before and during infusion of 0.2 mg.kg-1.min-1 adenosine i.v.. In 12 dogs, infarct size was measured at this time. In the remaining 9 dogs, reperfusion was continued for 3 hours, when MCE was repeated before and after an infusion of 0.56 mg.kg-1.min-1 dipyridamole i.v. and infarct size was measured. In the absence of adenosine, MCE perfusion defect at 15 minutes underestimated infarct sizes at both 15 minutes and 3 hours, whereas in the presence of adenosine, the estimate of infarct size was more accurate. Similarly, in the absence of dipyridamole, although MCE perfusion defect underestimated infarct size (both measured 3 hours after reflow), in the presence of dipyridamole, the estimate of infarct size was more accurate. CONCLUSIONS: By unmasking abnormalities in flow reserve within the infarct bed, MCE in conjunction with coronary vasodilators can accurately predict infarct size both 15 minutes and 3 hours after reperfusion. Thus, MCE can be used for assessing the extent of myocardial viability both immediately and late after reperfusion when postreflow coronary hyperemia is still present.


Assuntos
Circulação Coronária , Ecocardiografia , Infarto do Miocárdio/diagnóstico por imagem , Infarto do Miocárdio/fisiopatologia , Miocárdio/patologia , Adenosina/farmacologia , Animais , Dipiridamol/farmacologia , Cães , Coração/efeitos dos fármacos , Coração/fisiopatologia , Infarto do Miocárdio/patologia , Reperfusão Miocárdica , Análise de Regressão , Fatores de Tempo , Vasodilatação
20.
J Am Soc Echocardiogr ; 8(4): 453-64, 1995.
Artigo em Inglês | MEDLINE | ID: mdl-7546781

RESUMO

This study was designed to answer the question of whether, despite their theoretic superiority, integrated backscatter imaging (IBS) and digital data acquisition (DA) offer any advantage over conventional echocardiography (CE) during quantitative myocardial contrast echocardiography. In vitro experiments were performed (1) to determine the microbubble concentration versus videointensity relationships for CE and IBS and (2) to define the relationship between flow through and microbubble transit rates for CE and IBS. These data were stored on videotape. In vivo experiments were performed whereby microbubbles were injected into the left anterior descending artery at different flow rates in 14 dogs and IBS and CE data were stored both in digital format and on videotape. Although the level of compression did not affect the microbubble concentration versus videointensity plots during IBS compared with CE, in practical terms the mean transit rate, peak intensity, and area under the curve were not affected by the level of compression for both forms of imaging as long as the postprocessing used for CE imaging was linear and the microbubble dose was small. In addition, although DA resulted in higher peak intensity and area under the curve compared with storage on videotape because of its broader dynamic range, the correlation between these measurements was excellent with both forms of image storage. We conclude that, although differences exist between CE and IBS and between Da and analog acquisition, these differences do not significantly affect the derivation of parameters from time-intensity plots during myocardial contrast echocardiography when contrast material is injected into a coronary artery.


Assuntos
Ecocardiografia/métodos , Animais , Vasos Coronários , Cães , Processamento de Imagem Assistida por Computador , Injeções , Processamento de Sinais Assistido por Computador , Gravação em Vídeo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...