Correlação e concordância entre medidas ecocardiográficas obtidas durante o exame no ecocardiógrafo com medidas de imagens digitalizadas: estudo transversal

OBJETIVO: Avaliar a correlação e concordância entre medidas ecocardiográficas das dimensões cardíacas, obtidas através do aplicativo Echo off-line (programa para obtenção de medidas de imagens digitalizadas em estação de trabalho dedicada), com as realizadas convencionalmente. MÉTODOS: Estudo transversal, contemporâneo, sendo randomizados 56 pacientes. Através do programa Echo off-line foram mensuradas as medidas ao modo M e 2D ao nível dos ventrículos, do átrio esquerdo e da aorta. Estas medidas foram comparadas às realizadas por outro profissional, através do teste de correlação de Pearson (r), com alfa crítico de 0,05 e pela análise de concordância (Bland e Altman). RESULTADOS: As mensurações realizadas no sistema Echo off-line demonstraram r de 0,85 a 0,98. A análise de concordância mostrou que, para a maioria das medidas, a diferença média entre os métodos foi aproximadamente zero. A variação de valores absolutos não apresentou, em média, significância clínica. O aplicativo Echo off-line permite uma redução de, aproximadamente, 30% no tempo para realização das medidas. CONCLUSÃO: Este trabalho demonstrou a acurácia do programa Echo off-line para mensurar as dimensões cardíacas em estação de trabalho dedicada, podendo ser utilizado rotineiramente nos laboratórios de ecocardiografia.

Índice de dissincronia ventricular: comparação com a fração de ejeção bidimensional e tridimensional

FUNDAMENTO: O acoplamento eletromecânico (sincronia) do ventrículo esquerdo (VE) tem importância na análise da performance sistólica, especialmente para a indicação da terapia de ressincronização cardíaca em pacientes com ICC avançada. OBJETIVO: Comparar a sincronia do VE analisada com ecocardiograma (eco) tridimensional (3D) em tempo real com medidas de FEVE obtidas com ECO 2D e 3D. MÉTODOS: Estudo prospectivo de 92 indivíduos (56 homens, 47±10 anos), 60 com anatomia cardíaca (eco) e ECG normais (Grupo N), 32 com cardiomiopatia dilatada (Grupo CMD). Com o emprego do ECO 3D foram aferidos FEVE, volumes e índice de dissincronia (ID)% para 16 segmentos do VE; com o ECO 2D foram medidos FEVE (método de Simpson) e volumes sistólico e diastólico do VE. Análise estatística: coeficiente de correlação (Pearson), 95% IC, teste de regressão linear, teste de Bland & Altman, p<0,05. RESULTADOS: O ID% variou de 0,2900 a 28,1000 (5,2014±6,3281), a FEVE 3D variou de 0,17 a 0,81 (0,52±0,17); a FEVE 2D variou de 0,3 a 0,69 (0,49±0,11). A correlação entre ID e FEVE 3D foi (r): -0,7432, p<0,0001, IC: -0,8227 a -0,6350, a relação linear entre ID (x) e FEVE 3D (y) foi y = 19,8124 + (-27,9578) x , p<0,0001. A correlação entre ID e FEVE 2D foi (r): -0,7012, p<0,0001, IC: -0,7923 a -0,5797. CONCLUSÃO: Nesta casuística foi observada boa correlação negativa entre o acoplamento sistólico tridimensional eletromecânico do VE e a FEVE medida ao ecocardiograma (3D e 2D).

Parâmetros ecocardiográficos de deposição de gordura epicárdica e sua relação com doença arterial coronariana

FUNDAMENTO: Gordura epicárdica tem sido associada com a presença de doença arterial coronariana (DAC) significante. Entretanto, a associação entre infiltração lipomatosa do septo atrial e a infiltração de gordura do ventrículo direito (VD) permanece incerta. Nenhum desses parâmetros foi totalmente estudado em pacientes hispânicos. OBJETIVO: Determinar a associação entre a gordura epicárdica, infiltração lipomatosa do septo atrial e a infiltração de gordura do VD na presença de DAC. MÉTODOS: Duzentos e cinquenta pacientes hispânicos (86 mulheres e 164 homens, média da idade 61,5 ± 8 vs 62 ± 10 respectivamente), submetidos à sua primeira angiografia coronariana invasiva (ACI) foram estudados. No dia seguinte ao ACI, parâmetros de deposição de gordura epicárdica foram avaliados através de ecocardiografia modo 2D. Variáveis clínicas (idade, sexo, antecedentes pessoais do hábito de fumar, hipertensão e diabete melito, bem como a apresentação clínica da DAC) e antropométricas (circunferência da cintura e índice de massa corporal [IMC]) também foram coletadas. RESULTADOS: A gordura epicárdica (OR 1,27 p = 0,009), bem como a infiltração de gordura no VD (OR 2,94 p = 0,027), apresentaram uma associação significante e independente com a presença, mas não com a extensão (p = 0,516) e apresentação clínica (p = 0,153) da DAC. A extensão da deposição da gordura epicárdica mostrou uma associação proporcional e significante (p = 0,001) com a presença de DAC. CONCLUSÃO: A Gordura epicárdica e a infiltração de gordura do VD foram fatores significantes e independentes associados com a presença de DAC, a qual estava proporcionalmente aumentada de acordo com a extensão da deposição de gordura epicárdica.

Estudo da Mecânica Cardíaca pelo Speckle Tracking

Speckles, ou marcadores naturais do miocárdio, originam se da interferência construtiva e destrutiva do feixe de ultrassom que incide sobre os tecidos, podem fornecer um diagnóstico precoce das alterações miocárdicas e atuar na predição de certos eventos cardíacos. Devido à sua relativa estabilidade temporal, os speckles podem ser rastreados durante o ciclo cardíaco por software dedicados, promovendo a análise da função sistólica e diastólica. São identificados tanto pela escala de cinza da ecocardiografia 2D convencional quanto pela ecocardiografia 3D, sendo independentes do ângulo de incidência do ultrassom, permitindo assim a avaliação da mecânica cardíaca nos três planos espaciais: longitudinal, circunferencial e radial. O objetivo do presente artigo é discutir o papel e o significado da deformação cardíaca obtida por meio do speckle tracking durante a avaliação da fisiologia cardíaca, e discutir as aplicações clínicas desta tecnologia ecocardiográfica inovadora.

A Comparison of Radiation Dose Between Standard and 3D Angiography in Congenital Heart Disease

Background: The use of three-dimensional rotational angiography (3D-RA) to assess patients with congenital heart diseases appears to be a promising technique despite the scarce literature available. Objectives: The objective of this study was to describe our initial experience with 3D-RA and to compare its radiation dose to that of standard two-dimensional angiography (2D-SA). Methods: Between September 2011 and April 2012, 18 patients underwent simultaneous 3D-RA and 2D-SA during diagnostic cardiac catheterization. Radiation dose was assessed using the dose-area-product (DAP). Results: The median patient age and weight were 12.5 years and 47.5 Kg, respectively. The median DAP of each 3D-RA acquisition was 1093µGy.m2 and 190µGy.m2 for each 2D-SA acquisition (p<0.01). In patients weighing more than 45Kg (n=7), this difference was attenuated but still significant (1525 µGy.m2 vs.413µGy.m2, p=0.01). No difference was found between one 3D-RA and three 2D-SA (1525µGy.m2 vs.1238 µGy.m2, p = 0.575) in this population. This difference was significantly higher in patients weighing less than 45Kg (n=9) (713µGy.m2 vs.81µGy.m2, P = 0.008), even when comparing one 3D-RA with three 2D-SA (242µGy.m2, respectively, p<0.008). 3D-RA was extremely useful for the assessment of conduits of univentricular hearts, tortuous branches of the pulmonary artery, and aorta relative to 2D-SA acquisitions. Conclusions: The radiation dose of 3D-RA used in our institution was higher than those previously reported in the literature and this difference was more evident in children. This type of assessment is of paramount importance when starting to perform 3D-RA.

Right Ventricular Doppler Echocardiographic Study of Indeterminate Form of Chagas Disease

Background: Patients with indeterminate form of Chagas disease/cardiac normality (ICD/CN) exhibited normal electrocardiograms and chest X-rays; however, more sophisticated tests detected some degree of morphological and functional changes in the heart. Objective: To assess the prevalence of systolic and diastolic dysfunction of the right ventricle (RV) in patients with ICD/CN. Methods: This was a case–control and prevalence study. Using Doppler two-dimensional echocardiography (2D), 92 patients were assessed and divided into two groups: group I (normal, n = 31) and group II (ICD/CN, n = 61). Results: The prevalence of RV systolic dysfunction in patients in groups I and II was as follows: fractional area change (0.0% versus 0.6%), mobility of the tricuspid annulus (0.0% versus 0.0%), and S-wave tissue Doppler (6.4% versus 26.0%, p = 0.016). The prevalence of global disorders such as the right myocardial performance index using tissue Doppler (16.1% versus 27.8%, p = 0.099) and pulsed Doppler (61.3% versus 68%, p = 0.141) and diastolic disorders such as abnormal relaxation (0.0% versus 6.0%), pseudonormal pattern (0.0% versus 0.0%), and restrictive pattern (0.0% versus 0.0%) was not statistically different between groups. Conclusion: The prevalence of RV systolic dysfunction was estimated to be 26% (S wave velocity compared with other variables), suggesting incipient changes in RV systolic function in the ICD/CN group.

Subclinical Ventricular Dysfunction Detected by Speckle Tracking Two Years after Use of Anthracycline

Background: Heart failure is a severe complication associated with doxorubicin (DOX) use. Strain, assessed by two-dimensional speckle tracking (2D-STE), has been shown to be useful in identifying subclinical ventricular dysfunction. Objectives: a) To investigate the role of strain in the identification of subclinical ventricular dysfunction in patients who used DOX; b) to investigate determinants of strain response in these patients. Methods: Cross-sectional study with 81 participants: 40 patients who used DOX ±2 years before the study and 41 controls. All participants had left ventricular ejection fraction (LVEF) ≥55%. Total dose of DOX was 396mg (242mg/ms2). The systolic function of the LV was evaluated by LVEF (Simpson), as well as by longitudinal (εLL), circumferential (εCC), and radial (εRR) strains. Multivariate linear regression (MLR) analysis was performed using εLL (model 1) and εCC (model 2) as dependent variables. Results: Systolic and diastolic blood pressure values were higher in the control group (p < 0.05). εLL was lower in the DOX group (-12.4 ±2.6%) versus controls (-13.4 ± 1.7%; p = 0.044). The same occurred with εCC: -12.1 ± 2.7% (DOX) versus -16.7 ± 3.6% (controls; p < 0.001). The S’ wave was shorter in the DOX group (p = 0.035). On MLR, DOX was an independent predictor of reduced εCC (B = -4.429, p < 0.001). DOX (B = -1.289, p = 0.012) and age (B = -0.057, p = 0.029) were independent markers of reduced εLL. Conclusion: a) εLL, εCC and the S’ wave are reduced in patients who used DOX ±2 years prior to the study despite normal LVEF, suggesting the presence of subclinical ventricular dysfunction; b) DOX was an independent predictor of reduced εCC; c) prior use of DOX and age were independent markers of reduced εLL.

Studies on bioactive components from Chinese medicinal plants

Several novel bioactive components isolated from Chinese medicinal plants will be presented. These include novel maytansinoid tumor, inhibitors, some new ent-kaurane and rosane diterpenoids from Mallotus anomalus Meer et Chun (Euphorbiaceae), as well asnovel insecticide, stemona alkaloids from Stemona parviflora C. H. Wright (Stemonaceae). Both are native plants of Hainan island, Chine. 2D NMR techniques such as mono and hetero-COSY, NOESY, COLOC as well as H-NMR line broadening effect were utilized for structure elucidation. The separation techniques, struture elucidations and bioassay results will be reported.

Separation and Mapping of Chromosomes of Parasitic Protozoa

Many protozoan parasites represent an important group of human pathogens. Pulsed Field Gradient Gel Electrophoresis (PFGE) analysis has been an important tool for fundamental genetic studies of parasites like Trypanosoma, Leishmania, Giardia or the human malaria parasite Plasmodium falciparum. We present PFGE conditions allowing a high resolution separation of chromosomes ranging from 500 to 4000 kb within a two day electrophoresis run. In addition, we present conditions for separating large chromosomes (2000-6000 kb) within 36 hr. We demontrate that the application of two dimentional PFGE (2D-PFGE) technique to parasite karyotypes is a very useful method for the analysis of dispersed gene families and comparative studies of the intrachomosomal genome organization

Trypanosoma cruzi: Maintenance in Culture Modify Gene and Antigenic Expression of Metacyclic Trypomastigotes

In this study we examined whether the maintenance of Trypanosoma cruzi by long-time in axenic culture produces changes in gene expression and antigenic profiles. The studies were made with a Dm30L-clone from a low-virulent strain and a non-cloned virulent EP-strain of T. cruzi. Both parasites were maintained, for at least seven years, by successive alternate passage triatomine/mouse (triatomine condition), or by serial passage in axenic medium (culture condition). The comparison of the [35S]methionine metabolic labeling products of virulent and non-virulent parasites by 2D-SDS-PAGE, clearly indicates that the expression of metacyclic trypomastigotes (but not of epimastigotes) proteins have been altered by laboratory maintenance conditions. Western blot analysis of EP and Dm30L-epimastigotes using a serum anti-epimastigotes revealed that although most of antigens are conserved, four antigens are characteristics of triatomine condition parasites and three other are characteristics of culture condition parasites. Anti-metacyclics serum revealed significative differences in EP- and Dm30L-metacyclic trypomastigotes from triatomine condition. However, avirulent metacyclic forms were antigenically very similar. These results suggest that besides a possible selection of avirulent subpopulation from T. cruzi strains genetically heterogeneous when maintained by long time in axenic culture, changes in virulence might be due to post-translational modifications of the antigens induced by the absence of the natural alternability (vertebrate-invertebrate) in the life-cycle of T. cruzi

In Vitro and in Vivo Assays of 3,5-Disubstituted-Tetrahydro-2H-1,3,5-Thiadiazin-2-Thione Derivatives against Trypanosoma cruzi

Cytotoxicity assays of 24 new 3,5-disubstituted-tetrahydro-2H-1,3,5-thiadiazin-2-thione derivatives were performed. The 17 compounds with higher anti-epimastigote activity and lower cytotoxicity were, thereafter, screened against amastigote of Trypanosoma cruzi. Out of these 17 derivatives S-2d was selected to be assayed in vivo, because of its remarkable trypanocidal properties. To determine toxicity against J774 macrophages, a method based on quantification of cell damage, after 24 h, was used. Cell respiration, an indicator of cell viability, was assessed by the reduction of MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] to formazan. Anti-amastigote activity was estimated after 48 h by microscopic counts of May Grünwald-Giemsa-stained monolayers. Nifurtimox and benznidazole were used as reference drugs. For the in vivo experiences, mice were infected with 10(4) blood trypomastigotes and then treated during 15 days with S-2d or nifurtimox by oral route. All of the compounds were highly toxic at 100 µg/ml for macrophages and a few of them maintained this cytotoxicity even at 10 µg/ml. Of the derivatives assayed against amastigotes 3k and S-2d showed an interesting activity, that was held even at 1µg/ml. It is demonstrated that the high anti-epimastigote activity previously reported is mainly due to the non-specific toxicity of these compounds. In vivo assays assessed a reduction of parasitemia after administration of S-2d to infected mice.

From genomes to vaccines via the proteome

An effective vaccine against schistosomiasis mansoni would be a valuable control tool and the high levels of protection elicited in rodents and primates by radiation-attenuated cercariae provide proof of principle. A major obstacle to vaccine development is the difficulty of identifying the antigens that mediate protection, not least because of the size of the genome at 280Mb DNA encoding 14,000 to 20,000 genes. The technologies collectively called proteomics, including 2D electrophoresis, liquid chromatography and mass spectrometry, now permit any protein to be identified provided there is extensive DNA data, and preferably a genome sequence. Applied to soluble (cytosolic) proteins from schistosomes, proteomics reveals the great similarity in composition between life cycle stages, with several WHO vaccine candidates amongst the most abundant constituents. The proteomic approach has been successfully applied to identify the secretions used by cercaria to penetrate host skin, the gut secretions of adult worms and the proteins exposed on the tegument surface. Soluble proteins can also be separated by 2D electrophoresis before western blotting to identify the full range of antigenic targets present in a parasite preparation. The next step is to discover which target proteins represent the weak points in the worm's defences.

Crescimento micelial e síntese de proteínas de choque térmico em três isolados de fungos ectomicorrízicos sob condições de temperaturas supra-ótimas

A síntese de proteínas de choque térmico é uma alteração fisiológica transiente na célula de organismos expostos a temperaturas supra-ótimas. A resposta fisiológica ao choque térmico é dependente, particularmente, do tipo de célula e da capacidade dos organismos em responder às alterações do meio. O presente trabalho teve como objetivo avaliar o crescimento micelial e a síntese de proteínas de choque térmico de dois isolados de Pisolithus sp. (RV82 e RS24) e de um isolado de Paxillus involutus em temperaturas supra-ótimas. No trabalho, foram feitas análises de crescimento micelial em placa de Petri com meio apropriado para o crescimento sob condições de temperaturas subletais, letais e de choque térmico. As proteínas nos micélios dos isolados foram marcadas com aminoácido radioativo (³H-leucina), e a radioatividade, quantificada em solução de cintilação. A síntese das proteínas de choque térmico (HSPs) foi avaliada em gel de poliacrilamida (SDS-PAGE e 2D-PAGE). Demonstrou-se, com ³H-leucina, que os fungos ectomicorrízicos apresentaram respostas diferenciadas em relação ao crescimento micelial quando expostos a temperaturas supra-ótimas. Os dois isolados de Pisolithus sp., RS24 e RV82, mostraram-se mais tolerantes a altas temperaturas, quando comparado ao isolado de P. involutus. Os isolados de Pisolithus sp. diferem quanto à síntese de proteínas de estresse, com a síntese de HSPs de alta e de baixa massa molecular. Em resposta ao choque térmico, o isolado RV82 sintetizou proteínas putativas dos grupos HSP70, HSP28 e HSP26 e as sHSPs (15-18 kDa), enquanto o isolado RS24 sintetizou as dos grupos HSP86, HSP60, HSP55 e HSP35 e as sHSPs (12-18 kDa). A baixa tolerância a temperaturas elevadas do isolado de P. involutus foi atribuída à ausência de síntese de proteínas putativas do grupo HSPs em condições de choque térmico. Os resultados sugerem que os isolados de fungos ectomicorrízicos diferem quanto ao mecanismo de indução de termotolerância.

Modeling water movement in horizontal columns using fractal theory

Fractal mathematics has been used to characterize water and solute transport in porous media and also to characterize and simulate porous media properties. The objective of this study was to evaluate the correlation between the soil infiltration parameters sorptivity (S) and time exponent (n) and the parameters dimension (D) and the Hurst exponent (H). For this purpose, ten horizontal columns with pure (either clay or loam) and heterogeneous porous media (clay and loam distributed in layers in the column) were simulated following the distribution of a deterministic Cantor Bar with fractal dimension H" 0.63. Horizontal water infiltration experiments were then simulated using Hydrus 2D software. The sorptivity (S) and time exponent (n) parameters of the Philip equation were estimated for each simulation, using the nonlinear regression procedure of the statistical software package SAS®. Sorptivity increased in the columns with the loam content, which was attributed to the relation of S with the capillary radius. The time exponent estimated by nonlinear regression was found to be less than the traditional value of 0.5. The fractal dimension estimated from the Hurst exponent was 17.5 % lower than the fractal dimension of the Cantor Bar used to generate the columns.

Genetic associations in the detection of QTLs for wheat spike-related traits

The objective of this work was to assess the genetic diversity and population structure of wheat genotypes, to detect significant and stable genetic associations, as well as to evaluate the efficiency of statistical models to identify chromosome regions responsible for the expression of spike-related traits. Eight important spike characteristics were measured during five growing seasons in Serbia. A set of 30 microsatellite markers positioned near important agronomic loci was used to evaluate genetic diversity, resulting in a total of 349 alleles. The marker-trait associations were analyzed using the general linear and mixed linear models. The results obtained for number of allelic variants per locus (11.5), average polymorphic information content value (0.68), and average gene diversity (0.722) showed that the exceptional level of polymorphism in the genotypes is the main requirement for association studies. The population structure estimated by model-based clustering distributed the genotypes into six subpopulations according to log probability of data. Significant and stable associations were detected on chromosomes 1B, 2A, 2B, 2D, and 6D, which explained from 4.7 to 40.7% of total phenotypic variations. The general linear model identified a significantly larger number of marker-trait associations (192) than the mixed linear model (76). The mixed linear model identified nine markers associated to six traits.