Calcium-induced changes in the location and conformation of troponin in skeletal muscle thin filaments. (25/5118)

Troponin is the regulatory protein of striated muscle. Without Ca2+, the contraction of striated muscle is inhibited. Binding of Ca2+ to troponin activates contraction. The location of troponin on the thin filaments and its relation to the regulatory mechanism has been unknown, though the Ca2+-induced dislocation of tropomyosin has been studied. By binding troponin(C+I) to actin in an almost stoichiometric ratio and reconstituting actin-tropomyosin-troponin(C+I) filaments, we reconstructed the three-dimensional structure of actin-tropomyosin-troponin(C+I) with or without Ca2+ from electron cryomicrographs to about 2.5 or 3 nm resolution, respectively. Without Ca2+, the three-dimensional map reveals the extra-density region due to troponin(C+I), which extends perpendicularly to the helix axis and covers the N-terminal and C-terminal regions of actin. In the presence of Ca2+, the C-terminal region of actin became more exposed, and troponin(C+I) became V-shaped with one arm extending towards the pointed end of the actin filament. This structure can be considered to show the location of troponin(C+I) in at least one of the states of skeletal muscle thin filaments. These Ca2+-induced changes of troponin(C+I) provide a clue to the regulatory mechanism of contraction.  (+info)

Contrast-enhanced transthoracic second harmonic echo Doppler with adenosine: a noninvasive, rapid and effective method for coronary flow reserve assessment. (26/5118)

OBJECTIVES: The purpose of this study is to evaluate the feasibility in detecting blood flow in the left anterior descending coronary artery (LAD) using transthoracic color Doppler (CD) imaging (in both second harmonic and fundamental mode) along with contrast enhancement and to verify if this new noninvasive method along with adenosine is safe, rapid and effective in assessing coronary flow reserve (CFR). BACKGROUND: Feasibility of contrast-enhanced transthoracic Doppler recording (in both second harmonic and fundamental mode) of blood flow velocity in the LAD has not been assessed. Adenosine has a greater vasodilator potency and more favorable kinetics than dipyridamole and thus it can be more suitable for assessing CFR in conjunction with this method. METHODS: Sixty-one patients with angiographically patent LAD underwent CD (both in fundamental and harmonic mode) as well as color-guided pulsed wave (PW) Doppler recording of blood flow velocity in the distal LAD before and after intravenous contrast injection. A second group of patients (n = 77), undergoing coronary angiography, was submitted to transthoracic contrast-enhanced PW Doppler recording of blood flow velocity in the LAD using harmonic CD as a guide, at rest and during adenosine-induced hyperemia. RESULTS: Harmonic CD along with echo contrast consistently improved blood flow detection in the LAD; the success rate in detecting flow of optimal quality was 88% with this approach, whereas it was 11% and 16% with CD in fundamental mode, respectively, before and after contrast. Pulsed wave Doppler results paralleled those of harmonic CD (p < 0.001 contrast harmonic vs. fundamental). In the second group of patients coronary angiography revealed 0% to <40% stenosis in 24 patients (group I), > or =40% to < or =75% in 17 patients (group II) and >75% stenosis in 34 patients (group III). There was a significant difference in CFR among the three groups of patients; CFR for peak diastolic velocity was (mean +/- SD): 2.88+/-0.7 (group I), 2.09+/-0.5 (group II) and 1.51+/-0.5 cm/s (group II) (p < 0.05 group I vs. both group II and group III; p < 0.05 group II vs. group III). The whole examination took less than 10 min. CONCLUSIONS: Contrast-enhanced second harmonic Doppler recording of blood velocity in the LAD is highly feasible and in combination with adenosine it is a rapid, safe and effective method for assessing CFR ratio.  (+info)

Echo-enhanced Doppler sonography of focal nodular hyperplasia of the liver. (27/5118)

Lesions of focal nodular hyperplasia are hypervascular, benign focal liver lesions whose differentiation from other focal liver lesions is of significant clinical relevance. The purpose of this study was to investigate the echo-enhancing agent SHU 508A (Levovist) in the evaluation of focal nodular hyperplasia with Doppler sonography. We examined 49 patients with 71 lesions of focal nodular hyperplasia in the liver with gray scale and power Doppler sonography. In all patients Levovist was administered intravenously in a concentration of 300 to 400 mg galactose per milliliter. Visualization of the feeding vessels and the vascularity of the lesions were evaluated, and the resistive indices in the feeders and the hepatic arteries were assessed. In comparison with unenhanced power Doppler sonography, echo-enhanced power Doppler sonography yields a higher sensitivity in the detection of the feeding artery (97% versus 82%) in focal nodular hyperplasia and in the depiction of the radial vascular architecture in such lesions, especially those located in the left lobe of the liver. Lesions less than 3 cm in diameter do not consistently show a characteristic vascular architecture with echo-enhanced Doppler sonography. The resistive index of the tumor-feeding artery (mean, 0.51 +/- 0.09) is significantly (P < 0.0001) lower than that of the hepatic artery (mean, 0.65 +/- 0.06) and decreases as the size of the focal nodular hyperplasia increases. The administration of Levovist may improve the signal-to-noise ratio and thus visualization of the vascular architecture in focal nodular hyperplasia. Lesions located in the left lobe of the liver, which commonly are subject to disturbing motion artifacts in color Doppler sonography, will significantly benefit from the administration of Levovist. Echo-enhanced power Doppler sonography, however, is not capable of depicting a characteristic vascular pattern in small (< or = 3 cm) lesions of focal nodular hyperplasia that would guarantee a specific diagnosis.  (+info)

Confocal microscopy in biomedical research. (28/5118)

Confocal microscopy has allowed a major advance in biological imaging, since it represents a rapid, cost effective means of ecamining thick tissue specimens. In most cases, this involves fluorescence imaging and it is increasingly being used as a basic tool in biomedical research. Confocal microscopy allows the collection of thin optical sections, without the need for physical sectioning of the tissue. Additionally, confocal microscopes can usually produce images with greater sensitivity, contrast and resolution than those produced with normal light microscopes. We attempt to explain how this technology might be better used as a routine research tool. Since high quality, in-focus optical sections of thick tissue preparations can be generated quickly, confocal microscopy, in combination with immunofluorescence histochemistry, can now be used to examine complex three-dimensional distributions of distinct structures within tissues such as nerves within airways. Additionally, ultraviolet confocal microscopy allows the assessment of both dynamic and static phenomena in living cells and tissues. Thus, in addition to the imaging of fluorescence associated with structural elements, confocal microscopes can be used to quantitatively evaluate the distribution and fluxes of intracellular ions like calcium. Rapid, line-scanning confocal microscopes can be used in the assessment of dynamic events. For example, the in vivo imaging of microvascular permeability in airways becomes possible for the first time. By providing examples of some of our uses for confocal microscopy, we might encourage others to explore this relatively new and important texhnology for examining events and structures in single cells, tissue samples and in intact animals.  (+info)

Early homogeneously enhancing hemangioma versus hepatocellular carcinoma: differentiation using quantitative analysis of multiphasic dynamic magnetic resonance imaging. (29/5118)

The aim of this study was to determine the usefulness of quantitative analysis of multiphasic dynamic contrast-enhanced magnetic resonance (MR) imaging in differentiating early homogeneously enhancing hemangiomas from hepatocellular carcinomas (HCCs). Four-phased dynamic MR imaging at 10 sec (first phase of dynamic contrast-enhanced imaging, P1), 35 sec (second phase, P2), 60 sec (third phase, P3) and 300 sec (delay phase, P4) immediately after intravenous administration of 0.1 mmol/kg Gadolinium-DTPA was obtained with 1.5-T unit with breath-hold multisection FLASH (fast low angle-shot) sequence (TR/TE, 113-130 msec/4.1 msec; flip angle, 80 degrees). Thirty-three HCCs and 18 hemangiomas, homogeneously enhanced on P1, were included in the study. The images were evaluated quantitatively (SNR, signal-to-noise ratio; and CNR, contrast- to- noise ratio of lesions). Quantitatively, mean CNR was higher for hemangiomas than for HCCs on all phases, and the difference in CNRs between hemangioma and HCCs was statistically significant on P3 and P4 (p < 0.0001). When the cutoff for CNR was set at a value of 7.00 on P3 and 1.00 on P4, sensitivity, specificity and accuracy were 94.4%, 93.9%, and 94.1% on P3, and 94.4%, 81.8%, and 86.3% on P4, respectively. There was no statistically significant difference in SNRs between HCC and hemangioma. The differential diagnosis between early, homogeneously enhancing hemangiomas and HCCs was more confidently made with CNRs of lesions on P3 and P4 in dynamic contrast-enhanced MR imaging.  (+info)

Automated localisation of the optic disc, fovea, and retinal blood vessels from digital colour fundus images. (30/5118)

AIM: To recognise automatically the main components of the fundus on digital colour images. METHODS: The main features of a fundus retinal image were defined as the optic disc, fovea, and blood vessels. Methods are described for their automatic recognition and location. 112 retinal images were preprocessed via adaptive, local, contrast enhancement. The optic discs were located by identifying the area with the highest variation in intensity of adjacent pixels. Blood vessels were identified by means of a multilayer perceptron neural net, for which the inputs were derived from a principal component analysis (PCA) of the image and edge detection of the first component of PCA. The foveas were identified using matching correlation together with characteristics typical of a fovea-for example, darkest area in the neighbourhood of the optic disc. The main components of the image were identified by an experienced ophthalmologist for comparison with computerised methods. RESULTS: The sensitivity and specificity of the recognition of each retinal main component was as follows: 99.1% and 99.1% for the optic disc; 83.3% and 91.0% for blood vessels; 80.4% and 99.1% for the fovea. CONCLUSIONS: In this study the optic disc, blood vessels, and fovea were accurately detected. The identification of the normal components of the retinal image will aid the future detection of diseases in these regions. In diabetic retinopathy, for example, an image could be analysed for retinopathy with reference to sight threatening complications such as disc neovascularisation, vascular changes, or foveal exudation.  (+info)

Effect of software manipulation (Photoshop) of digitised retinal images on the grading of diabetic retinopathy. (31/5118)

AIMS: To determine whether software processing of digitised retinal images using a "sharpen" filter improves the ability to grade diabetic retinopathy. METHODS: 150 macula centred retinal images were taken as 35 mm colour transparencies representing a spectrum of diabetic retinopathy, digitised, and graded in random order before and after the application of a sharpen filter (Adobe Photoshop). Digital enhancement of contrast and brightness was performed and a X2 digital zoom was utilised. The grades from the unenhanced and enhanced digitised images were compared with the same retinal fields viewed as slides. RESULTS: Overall agreement in retinopathy grade from the digitised images improved from 83.3% (125/150) to 94.0% (141/150) with sight threatening diabetic retinopathy (STDR) correctly identified in 95.5% (84/88) and 98.9% (87/88) of cases when using unenhanced and enhanced images respectively. In total, five images were overgraded and four undergraded from the enhanced images compared with 17 and eight images respectively when using unenhanced images. CONCLUSION: This study demonstrates that the already good agreement in grading performance can be further improved by software manipulation or processing of digitised retinal images.  (+info)

Contrast-enhanced transcranial color-coded sonography in acute hemispheric brain infarction. (32/5118)

PURPOSE: The aim of the present study was to investigate the diagnostic potential of contrast-enhanced transcranial color-coded real-time sonography (CE-TCCS) in otherwise ultrasound-refractory acute stroke patients with an ischemia in the territory of the middle cerebral artery (MCA). Furthermore, correlations of CE-TCCS findings with clinical, angiographic, and CT results were investigated. METHODS: In 90 acute stroke patients with inadequate insonation conditions in unenhanced transcranial color-coded real-time sonography (TCCS) examinations, CE-TCCS, clinical, angiographic, and CT examinations were performed within 12 hours, 36 hours (CE-TCCS only), and 1 week after onset of clinical symptoms. A CT angiography (CTA) as reference method was available in 39 individuals. After application of a galactose-based echo-enhancing agent, the portion of conclusive ultrasound examinations of the MCA, as manifested by an MCA occlusion, decreased or increased flow velocity (FV), and symmetrical MCA FV, was evaluated. CE-TCCS findings on admission and during follow-up were correlated with infarction size as demonstrated on follow-up CT, and clinical findings were assessed by use of the European Stroke Scale. RESULTS: Adequate diagnosis was achieved in 74 of 90 patients (82%) by the use of echo contrast agents. MCA occlusion or reduction of MCA FV was found in 20 and 27 patients, respectively. MCA occlusion was confirmed by CTA in 17 cases. In one individual, false-positive diagnosis of MCA occlusion was made according to ultrasound criteria. In 5 patients with MCA occlusion, vessel recanalization was observed during follow-up; 15 of 27 patients with decreased flow velocities showed normalization after the third examination that was associated with a significantly better clinical outcome (P<0.0001). Furthermore, MCA occlusion or decreased FV in the first 12 hours were associated with significantly larger infarctions in the MCA territory compared with normal CE-TCCS findings (P<0.0001). CONCLUSIONS: CE-TCCS enables adequate diagnosis in approximately 80% of acute hemispheric stroke patients with insufficient unenhanced TCCS examinations. It is a reliable diagnostic tool regarding MCA mainstem and branch occlusions. Because this method conveys useful information concerning cerebral tissue and clinical prognosis, it may be useful to identify those patients who benefit most from local or intra-arterial thrombolytic therapy.  (+info)