Gd-DTPA-enhanced cranial MR imaging in children: initial clinical experience and recommendations for its use.

Gd-DTPA was administered prospectively to 65 consecutive children (ages 1 day to 18 years, mean 9.6 years) to document its utility and safety for routine cranial MR imaging. Precontrast T1- and T2-weighted scans and postcontrast T1-weighted scans were obtained. No complications or significant adverse reactions were encountered. Contrast enhancement was seen in 14 lesions from seven patients, but each of these patients had some abnormality also present on precontrast images. Contrast enhancement was thought to be extremely helpful in characterizing four primary tumors and moderately helpful in characterizing four other lesions. Absence of contrast enhancement was helpful in clarifying the nature of abnormalities seen in an additional four patients. Gd-DTPA may be used safely in children, but this study does not support its routine administration. The highest incremental diagnostic yield from its use will likely be among patients with suspected neoplasms or inflammatory diseases and among those requiring further characterization of lesions seen on precontrast scans.

Radioiodide imaging of struma cordis.

This is the first reported case in which struma cordis was demonstrated with radionuclide imaging. A 56-year-old white woman underwent surgical excision of a benign intracardiac thyroid mass (struma cordis). Subsequent radionuclide imaging with I-123 sodium iodide and Tc-99m labeled red blood cells demonstrated a normal cervical thyroid gland as well as a focus of activity in the mediastinum consistent with intracardiac thyroid.

Association of the epidermal growth factor receptor kinase with the detergent-insoluble cytoskeleton of A431 cells.

The epidermal growth factor receptor (EGF-R) on human epidermoid carcinoma cells, A431, was found to be predominantly associated with the detergent-insoluble cytoskeleton, where it retained both a functional ligand-binding domain and an intrinsic tyrosine kinase activity. The EGF-R was constitutively associated with the A431 cytoskeleton; this association was not a consequence of adventitious binding. The EGF-R was associated with cytoskeletal elements both at the cell surface, within intracellular vesicles mediating the internalization of the hormone-receptor complex, and within lysosomes. The EGF-R became more stably associated with cytoskeletal elements after its internalization. The cytoskeletal association of the EGF-R was partially disrupted on suspension of adherent cells, indicating that alteration of cellular morphology influences the structural association of the EGF-R, and that the EGF-R is not intrinsically insoluble. Cytoskeletons prepared from EGF-treated A431 cells, when incubated with gamma-32P-ATP, demonstrated enhanced autophosphorylation of the EGF-R in situ as well as the phosphorylation of several high molecular weight proteins. In this system, phosphorylation occurs between immobilized kinase and substrate. The EGF-R and several high molecular weight cytoskeletal proteins were phosphorylated on tyrosine residues; two of the latter proteins were phosphorylated transiently as a consequence of EGF action, suggesting that EGF caused the active redistribution of the protein substrates relative to protein kinases. The ability of EGF to stimulate protein phosphorylation in situ required treatment of intact cells at physiological temperatures; addition of EGF directly to cytoskeletons had no effect. These data suggest that the structural association of the EGF-R may play a role in cellular processing of the hormone, as well as in regulation of the EGF-R kinase activity and in specifying its cellular substrates.

Nerve growth factor- and epidermal growth factor-stimulated phosphorylation of a PC12 cytoskeletally associated protein in situ.

Nerve growth factor (NGF) and epidermal growth factor (EGF) produce stable alterations in PC12 cells that persist in the detergent-insoluble cytoskeleton, resulting in the phosphorylation of a 250,000-mol-wt cytoskeletally associated protein in situ. Treatment of PC12 cells with NGF or EGF, followed by detergent lysis of the cells and incubation of the resulting cytoskeletons with gamma-32P-ATP, permitted detection of hormonally stimulated, energy-dependent events, which result in the enhanced phosphorylation of a cytoskeletally associated protein as an immediate consequence of receptor occupancy. These events were elicited only upon treatment of intact cells at physiological temperatures. The NGF- and EGF-stimulated events occurred rapidly; however, they were a transient effect of hormone action. NGF and EGF were found to act through independent mechanisms to stimulate the in situ phosphorylation of the 250,000-mol-wt protein, as the effects of NGF, but not EGF, were blocked by methyltransferase inhibitors. The 250,000-mol-wt protein was phosphorylated on serine and threonine residues in response to both NGF and EGF although in somewhat different proportions. The data suggest that the hormone-stimulated labeling of the 250,000-mol-wt protein may be the result of either the direct activation of a protein kinase, the redistribution of the kinase relative to its substrates as a consequence of hormone action, or the coincident occurrence of these events.

The effects of chemical cross-linking agents on calcium-induced structural changes in skinned muscle fibers. Origin within thick filaments detected by optical diffraction methods.

We have reported earlier (Sabbadini, R.A., Rieser, G.D. and Paolini, P.J. (1979) Biochim. Biophys. Acta 578, 526-533) that physiological levels of calcium (pCa 6.95-5.49) can produce structural changes in thick filaments which are detectable as an intensity loss of the first-order optical diffraction lines from chemically skinned skeletal muscle fibers stretched beyond myofilament overlap. We now show that the calcium-induced intensity decrease results from structural changes within, rather than between, thick filaments. Glycerinated, detergent-treated fibers from frog semitendinosus muscle were incubated in 1-10 mM concentrations of dimethylsuberimidate (DMS), dithiobis(succinimidylpropionate) (DTSP) or dimethyl-3,3'-dithiobispropionimidate (DTBP) for 4 h. These substances are homobifunctional lysine-modifying cross-linking reagents known to restrict movement of S-1 heads and limit changes in the association of myosin rods within the core of the thick filament without affecting interfilament lattice spacing. Diffraction patterns from cross-linked cells in relaxing solution were identical to those in control cells, but Ca2+ (pCa 5.49) totally failed to produce the typical 50-70% attenuation of first-order line intensity. Cleavage of the disulfide bond in DTBP-treated cells with dithiothreitol fully restored the Ca2+ sensitivity. Lysine group modification with methylacetimidate, a monofunctional lysine modification reagent equivalent to DMS, did not block the Ca2+ sensitivity. We observed that intensity reductions can also be produced by numerous other agents and mechanisms, such as nonionic polymeric solutions of polyvinylpyrrolidone, which reduces the lattice spacing, and alkaline pH, which probably displaces the S-1 heads from a resting position close to the thick filament surface. However, the prevention of the Ca2+ effect by cross-linkers indicates that intrafilament rather than interfilament changes in structure are responsible for the light diffraction intensity decrease accompanying activation.

Calcium-induced structural changes in chemically skinned muscle fibers. Detection by optical diffractometry.

The intensity of the first order diffraction line produced by chemically skinned muscle fibers was detected by a self scanning photodiode array and minicomputer system. Line intensity was observed to decrease in fibers stretched to zero filament overlap when subjected to calcium-EGTA buffers in the physiological pCa range. Calcium dependent intensity decreases were not observed for myosin extracted fibers indicating that the thick filament proteins may be the source of the calcium effect seen in non-extracted fibers. These results can be interpreted in terms of calcium dependent effects on thick filament disordering which are not dependent upon cross bridge formation.