Polymorphonuclear leukocytes: possible mechanism of accumulation in psoriasis.

Extracts of involved and uninvolved skin from nine patients with untreated psoriasis were studied for chemotactic activity. Psoriatic plaque contains increased amounts of a complement-dependent chemotactic factor that is inhibited by diisopropyl fluorophosphate. This factor may be human skin serine proteinase.

Characterization of a chemotactic and cytotoxic proteinase from human skin.

A proteinase (EC 3.4.-.-) active at physiological pH has been isolated from human skin utilizing gel filtration and affinity chromatography techniques. The proteinase has a molecular weight of approx. 28 000 and it is inhibited by alpha 2-macroglobulin, alpha 1-antitrypsin, C-1 inactivatory, soybean trypsin inhibitor and diisopropyl fluorophosphate. 2njection of 1 ng of purified proteinase into rabbit skin induces polymorphonuclear leukocyte infiltration of the cutis. Inhibition of enzyme activity with diisopropyl fluorophosphate inhibits the chemotactic effect. Addition of 0.2 microgram/ml of purified proteinase to fibroblast cultures kills the cells within minutes. This proteinase may play a significant role in modulating the inflammatory response after cellular injury.

An affinity column method for determination of the immunoreactivity of 131I-chimeric L6 monoclonal antibody and comparison to in vivo tumor localization.

An affinity column method was developed to determine the immunoreactivity of 131I-ChiL6 (chimeric L6 monoclonal antibody), a candidate for radioimmunotherapy. This method involved assessing the binding of the radiolabeled antibody to antigen containing membranes bound to a Reacti-gel agarose matrix. The immunoreactivity determined by the affinity column method correlated to other in vitro binding assays including the Lindmo infinite antigen excess method. In tumor-bearing mice which had been injected with 131I-ChiL6, which possessed high immunoreactivities (90-82%), a high tumor uptake (13.5-10.5% ID/g) was observed. A decrease in tumor uptake (5.2-4.8% ID/g) was observed with 131I-ChiL6 samples of low immunoreactivity (42% and 31%, respectively). While a moderate loss of immunoreactivity (4-18%) of the 131I-ChiL6 samples could be detected by the affinity column method, the loss of tumor uptake in vivo observed was not as significant. This method was found to be an efficient and sensitive method for detecting damage to the antibody during radiolabeling and applicable as a quality control method for clinical trials. This rapid method, compared to the other in vitro binding assays (including the Lindmo infinite antigen excess method) has distinct advantages as a quality control method since it requires less manipulation and can be semi-automated.

Prekallikrein activation, C1 esterase inhibitor, and factor XII as predictors of adverse reaction to contrast media. A prospective study.

The susceptibility of 152 patients to idiosyncratic reactions resulting from the administration of radiographic contrast media was studied. The rate of activation of plasma prekallikrein was measured in samples taken from these patients before they received contrast agents. Kallikrein inhibitor and factor XII levels were also determined. The tests were of no value in selecting the ten patients who subsequently experienced mild reactions. However, the possibility remains that one or more of the tests may have predictive value for patients who experience moderate or severe reactions.

A predictive test for adverse reactions to contrast media. Preliminary results.

In a prospective study, whole blood samples drawn from patients prior to their being injected with contrast media were incubated with zymosan to activate the complement cascade. The samples were tested for various analytes, including C3a, thromboxane B2 (TxB2), beta thromboglobulin and platelet factor 4 (PF4). Of 207 patients receiving contrast media, only eight experienced reactions, which were mild. Levels of the platelet constituents were generally elevated in these patients. Specificity and sensitivity were 89% and 83%, respectively, for the combined TxB2 and PF4 radioimmunoassay data. Using the Wilcoxon-Mann-Whitney rank sum test, both PF4 and TxB2 were collected with RCM reactions at the R less than .05 level. Although preliminary, the results suggest that RCM reactions are predictable by the in vitro test procedures described.

Comparative chemical structure and pharmacokinetics of MRI contrast agents.

The blood clearance kinetics of five gadolinium complexes, Gd(L), were determined in rats and the results interpreted in terms of an open two-compartment pharmacokinetic model. The complexes were tested in vitro for stability in serum and in aqueous solutions of ions that they might encounter in vivo and that might be expected to react with the Gd(L) complexes to produce uncomplexed gadolinium. Reaction with serum was observed in two instances. Chemical structural differences among the chelating ligands appear to govern the overall reactivity of their Gd(L) complexes. It may be inferred from the results that a preferred structural feature of the ligand is the presence of a 12-membered 1,4,7,10-tetraaza macrocycle.

A specific radioimmunoassay for the measurement of gadoteridol, a contrast agent for magnetic resonance imaging in biological fluids.

Gadoteridol, a nonionic gadolinium chelate, is currently being evaluated for contrast-enhanced magnetic resonance imaging. A radioimmunoassay (RIA) has been developed for the measurement of gadoteridol in biological fluids. The RIA has a range of 0 to 25 micrograms/mL and has the sensitivity to detect 0.05 microgram/mL of gadoteridol. Satisfactory zero binding and sensitivity were obtained after an overnight incubation at 4 degrees C. Separation of the antibody-bound and free radiolabel was achieved with 12.5% polyethylene glycol. A quantitative recovery of the exogenous analyte was obtained at all concentrations of gadoteridol tested. Linearity in both serum and urine was satisfactory. Intraassay coefficients of variation were 6.4 and 2.8% for the low and medium controls, respectively. Interassay coefficients of variation were 5.4, 3.8, and 12.2% for the low, medium, and high controls, respectively. Cross reactivities of the ligand 5 and the calcium salt 6 were 37 and 29%, respectively. Clinical samples from the ascending dosage studies were analyzed by the gadoteridol RIA. The results obtained from the serum specimens demonstrated an excellent linear proportionality between drug concentration in blood and administered dosage of gadoteridol. Cumulative urinary excretion data showed that 94% of the drug was excreted in the urine within 24 h.

Superoxide dismutases of Escherichia coli: intracellular localization and functions.

Escherichia coli B contains two superoxide dismutases which differ with respect to their localization within the cell, the nature of their prosthetic metals, their responses to changes in (p)O(2), and their functions. One of these enzymes, which was liberated from the cells by osmotic shock and which was therefore presumed to be localized in the periplasmic space, is an iron-containing superoxide dismutase. The amount of this iron enzyme did not vary in response to changes in (p)O(2) during growth. In contrast, the other superoxide dismutase was not solubilized by osmotic shock, was a mangano-protein, and was found in greater amounts in cells which had been grown at high (p)O(2). E. coli, which had low levels of the iron-enzyme and high levels of the mangano-enzyme, as a consequence of growth in iron-deficient aerated medium, was killed by exposure to an exogenous flux of O(2) (-) which was generated either photochemically or enzymatically. The addition of bovine superoxide dismutase to the suspending medium protected these cells against this stress. On the other hand, E. coli, which had high levels of the iron-enzyme and low levels of the mangano-enzyme, as a consequence of growth in iron-rich anaerobic medium, was resistant to exogeneous O(2) (-). On the basis of these and of previously reported results (4a, Yost, F. J. and I. Fridovich, J. Biol. Chem., 1973, in press), it appears that the iron superoxide dismutase, of the periplasmic space, serves as a defense against exogenous O(2) (-), whereas the mangano-superoxide dismutase, in the matrix of these cells, serves to counter the toxicity of endogenous O(2) (-).

Production and secretion of proteolytic enzymes by normal and neoplastic cells.

A possible mechanism for tumor cell invasion of normal tissue might be secretion of proteolytic enzymes. This study compares and contrasts production and secretion of proteinases by cell cultures of normal and chemically transformed mouse epithelial cells. Lysates of normal and neoplastic cells contain similar amounts of neutral proteinase, cathepsin D and plasminogen activator. Neither collagenase nor elastase could be identified in lysates of, or serum-free culture medium bathing, normal or neoplastic cells. Neoplastic cells secrete ten times more plasminogen activator than normal cells. Our data support the hypothesis that plasminogen activator produced by neoplastic cells could fuction to activate latent proteolytic enzymes secreted by connective tissue cells which might result in spread of neoplastic cells into normal tissue.