Vinculin is an essential component for normal myofibrillar arrangement in fetal mouse cardiac myocytes

Vinculin is a cytoskeletal protein that is believed to be an essential component in the linkage of cytoskeletal actin filaments to the plasma membrane. To investigate the precise function of vinculin in the development of cardiac myofibrils, antisense oligodeoxynucleotides complementary to vinculin mRNA were used to perturb the expression of the protein during myofibril assembly and arrangement in mouse cardiac myocytes. Fetal (day 18-20 post-conception) mouse cardiac myocytes were isolated by collagenase digestion, separated by Percoll density gradient centrifugation, and plated on aligned collagen gels. By 72 h of culture, mouse myocytes displayed an elongated in vivo-like phenotype in parallel with the aligned fibrils of the collagen gels with polarized arrays of myofibrils. Two different antisense oligonucleotides (20-mer) altered the formation of the tissue-like phenotype of myocytes. These antisense oligonucleotides suppressed vinculin protein expression at 43.5+/-26.8% and 48.7+/-20.9% when compared to myocytes that were not treated. Examination of these myocytes by confocal scanning laser and transmission electron microscopy revealed a disruption of the aligned in vivo-like phenotype, assembly of thick and thin filaments, and formulation of Z-bands. Random sequence 20-mer oligonucleotides used as controls had little detectable effect on vinculin protein expression (94.2+/-14.8%), cell shape, normal alignment or assembly of myofibrils. These results indicate that vinculin is a critical cytoskeletal component, that functions in the determination of cell shape and the arrangement and organization of developing myofibrils.     

Detection of coronary stenoses on source and projection images using three-dimensional MR angiography with retrospective respiratory gating: preliminary experience

OBJECTIVE: Our objective was to study the ability of three-dimensional MR angiography with retrospective respiratory gating to reveal stenoses in proximal coronary arteries on source and projection images. CONCLUSION: Proximal coronary artery stenoses can be identified using three-dimensional MR angiography with retrospective respiratory gating, both with projection images and on source images alone. Reasons for missed lesions included collateral vessels and retrograde flow distal to complete occlusion and volume averaging of vessels with adjacent structures. Causes of false-positive interpretations included small foci of decreased signal intensity distal to complete occlusion, partial volume effects on individual partitions, and regions of distal vessels leaving the imaging plane.     

Geranoyl-CoA carboxylase: a novel biotin-containing enzyme in plants

Geranoyl-CoA carboxylase (EC 6.4.1.4) is a biotin-containing enzyme previously described in two genera of bacteria. Here we report the presence of geranoyl-CoA carboxylase in kingdom Plantae. Geranoyl-CoA carboxylase was purified 180-fold from maize leaves. The enzyme has a biotin-containing subunit of 122 kDa. The pH optimum for activity is 8.3. The apparent Km values for the substrates geranoyl-CoA, bicarbonate, and ATP are 64 +/- 5 microM, 0. 58 +/- 0.04 mM, and 8.4 +/- 0.4 microM, respectively. Subcellular fractionations indicate that geranoyl-CoA carboxylase is located in plastids. Geranoyl-CoA carboxylase activity is ubiquitous in organs of monocots and dicots and varies with development. We postulate that geranoyl-CoA carboxylase plays an important role in isoprenoid catabolism in plants, in a pathway analogous to that shown in Psuedomonas sp. In plants, this catabolic pathway would require the interaction of at least three subcellular compartments (plastids, microbodies, and mitochondria) and two biotin-containing enzymes, geranoyl-CoA carboxylase and 3-methylcrotonyl-CoA carboxylase.     

Experimental hepatobiliary fascioliasis in rabbits: a radiology-pathology correlation

RATIONALE AND OBJECTIVES: The authors sought to correlate the radiologic findings of hepatobiliary fascioliasis with pathologic features. METHODS: Serial ultrasound, computed tomography (CT), and magnetic resonance findings in seven rabbits with experimentally induced fascioliasis were obtained every other week. Direct cholangiogram was also obtained after the rabbits were killed. Radiology-pathology correlation was done in specimens. RESULTS: In the parenchymal phase (an acute phase of parenchymal invasion of a larva), CT showed subcapsular clustered areas of low attenuation. Magnetic resonance appearance was similar in shape but better than CT in characterizing the hemorrhagic nature of the lesion. Ultrasound findings were nonspecific in this phase. In the ductal phase (a stationary phase after residing in the bile duct), CT showed dilatation of central ducts with symmetric periportal hypoattenuation (periportal tracking). Magnetic resonance could not depict mild ductal dilatation. Ultrasound was most valuable in demonstrating the moving worm within the dilated duct. Pathologically, the hepatic parenchymal lesions consisted of a cluster of eosinophilic granulomas with hemorrhagic change (migratory tract of the flukes). Ductal changes were observed predominantly in the central bile ducts. Periportal lymphangiectasia was also noted. CONCLUSIONS: Computed tomography or magnetic resonance can demonstrate the characteristic evolutionary pattern of fascioliasis that reflects the unique life cycle of Fasciola hepatica. The role of ultrasound, although limited in the parenchymal phase, was most useful in the ductal phase in that it demonstrated the moving worms themselves.     

Preliminary core set of domains and reporting requirements for longitudinal observational studies in rheumatology

The minimized energy mapping of annexin V Trp-187 chi1 x chi2 isomerization supports the existence of two preferential rotameric orientations of the Trp side chain upon annexin V binding to membranes, in agreement with the time-resolved fluorescence results. They correspond to the perpendicular trans (-173 degrees, 73 degrees) and g- (-71 degrees, 83 degrees) rotamers and represent 59 and 28% of the population, respectively. The analysis of their local environment makes it possible to assign the trans rotamer to the long component and the g- rotamer to the short component of the biexponential fluorescence decay. The orientation of these rotamers relative to the protein core suggests a dual role for Trp-187, which might be involved both in the interaction with the phospholipid bilayer and in the formation of the annexin V 2-D array at the surface of the membrane.     

Automated detection of bolus arrival and initiation of data acquisition in fast, three-dimensional, gadolinium-enhanced MR angiography

Automatic triggering of magnetic resonance (MR) angiography with detection of a contrast material bolus was evaluated. Signal intensity changes with time were tracked in a prescribed tracking or monitoring volume by a parallel signal processing unit that automatically started data acquisition once user-defined thresholds were exceeded. This technique, referred to as MR Smartprep, was reliable and avoided the inconsistencies of manual timing.