A strategy for the evaluation of sensory and pulmonary irritation due to chemical emissions from indoor sources.

Sensory and pulmonary irritation are physiological responses to chemical exposure which result in characteristic, measurable changes in respiratory activity in mice. A standard method has been applied to the estimation of sensory irritation associated with a specific chemical exposure. This method has been correlated with human responses to these chemicals. Symptoms associated with chemical irritants are consistent with complaints due to problems with indoor air quality, which may include eye and upper respiratory tract irritation, headaches, and nausea. A stepwise strategy for assessing the contribution of indoor products to sensory and pulmonary irritation is discussed in the current paper. The strategy includes product emissions testing using dynamic environmental chambers, the selection of suspected irritants for respiratory irritation testing, respiratory irritation testing of individual compounds are representative mixtures using synthesized atmospheres, and the evaluation of test data to determine those compounds which may contribute to sensory and pulmonary irritation in humans. The current strategy is being applied to evaluate carpet system materials and their constituent chemicals.

Toxic epidermal necrolysis.

Toxic epidermal necrolysis is a potentially fatal dermatological disease. Large bullae covering extensive areas of the body cause continuous exfoliation of skin, which requires immediate medical attention. Intraoral manifestations may precede cutaneous lesions. Two cases with different treatment protocols are presented.

Immunohistochemical localization of glutamine transaminase K, a rat kidney cysteine conjugate beta-lyase, and the relationship to the segment specificity of cysteine conjugate nephrotoxicity.

Rat kidney glutamine transaminase K is a major rat kidney cysteine conjugate beta-lyase and is a key enzyme in the nephrotoxicity of some cysteine conjugates. However, it has not been demonstrated that the beta-lyase is present in the target cells. Furthermore, although all segments of the proximal tubule are affected by high doses of nephrotoxic cysteine conjugates, the S3 segment is the most sensitive. Because heterogeneous distribution of the beta-lyase could account for the enhanced sensitivity, antibody raised against rat kidney cysteine conjugate beta-lyase has been prepared and used to investigate the distribution of the enzyme in kidney and other tissues. The data show that the enzyme is highest in rat kidney, consistent with enzyme activity data. By immunohistochemical staining, no enzyme is present in the glomeruli or distal tubular elements of the kidney. The enzyme is present only in the target cells, the renal proximal tubular epithelium. However, the distribution of the beta-lyase within the proximal tubule is not consistent with the hypothesis that a higher concentration of the enzyme in the S3 segment accounts for the greater sensitivity of S3 to nephrotoxic cysteine conjugates compared to S1 and S2. Several alternative hypotheses are discussed.

Mechanism of transport for toxic cysteine conjugates in rat kidney cortex membrane vesicles.

Cysteine conjugate transport plays a key role in the interorgan transport of xenobitoic metabolites which are formed via the mercapturic acid pathway. In the rat, transport of cysteine conjugates could be an important factor in the selective nephrotoxicity of some toxic cysteine conjugates. However, little information is available on the molecular mechanism(s) of cysteine conjugate transport in the rat kidney. Therefore, we have investigated the polarity and the molecular driving forces for the transport of S-(1,2-dichlorovinyl)-L-cysteine (DCVC) in isolated membrane vesicles from rat kidney cortex. Our data suggest that Na+-dependent transport on the lumenal side is responsible for the uptake of cysteine conjugates across the apical membrane. No Na+-stimulated transport was found on the basolateral side and uptake of DCVC in basolateral membrane vesicles was not saturable. Na+-dependent transport in brush border membrane vesicles was inhibited by a variety of neutral amino acids and cysteine conjugates, but not by polar amino acids. Therefore, the transporter is similar to the Na+-dependent neutral amino acid transporter of rat kidney brush border membranes. The system L-specific substrate, 2-amino-2-norbornane carboxylic acid, was not inhibitory. The Km for the Na+-stimulated transport system in brush border membrane vesicles was 225 microM and the Vmax was 782 pmol/15 sec/mg of protein. We propose that the driving force for the apical transport of cysteine conjugates may be the coupling of the lumenal transport to the Na+-gradient. The data are discussed with regard to a transepithelial transport model for cysteine conjugates and the role transport plays in the molecular mechanism of cysteine conjugate toxicity.

The transport of S-cysteine conjugates in LLC-PK1 cells and its role in toxicity.

The transport of S-cysteine conjugates was studied in the kidney cell line, LLC-PK1, using the nephrotoxin, S-(1,2-dichlorovinyl)-L-cysteine (L-DCVC), as the model compound. The saturable uptake of this conjugate did not require sodium and was selectively inhibited by the amino acid transport system L-specific substrate, 2-amino-2-norbornane carboxylic acid, as well as a variety of other S-cysteine conjugates and neutral amino acids with large, nonpolar side chains. Kinetic studies suggested the existence of both low and high affinity transport systems with Km values that differed by 25-fold. Although these uptake systems showed no discernible differences in substrate specificity, the low affinity transport was more sensitive to trans-stimulation. L-DCVC uptake in subconfluent cultures was about 3-fold that of confluent cells, suggesting either adaptive regulation to cell growth or polarization of transport to the basolateral membrane. L-DCVC toxicity in LLC-PK1 cells was inhibited in the presence of nontoxic transport substrates but was potentiated when cells were preloaded with many of the same compounds, indicating that transport may be a rate-limiting factor in L-DCVC-induced toxicity under certain circumstances. The possible role of this system L-like uptake in the transport of S-cysteine conjugates in vivo is discussed.

Analysis of monobutyryl and dibutyryl derivatives of adenosine 3',5'-monophosphate in biological samples using isocratic ion pair high-performance liquid chromatography.

Adenosine 3',5'-monophosphate (cyclic AMP), its dibutyryl and monobutyryl derivatives, and a number of other naturally occurring adenine-containing compounds were separated by isocratic ion pair high-performance liquid chromatography. A mobile phase consisting of 30% methanol in 0.1 M KH2PO4 (pH 4.0) containing 1 mM tetramethylammonium hydroxide as the counterion was used to separate the butyryl derivatives. To sufficiently separate cyclic AMP from other adenine-containing compounds, a mobile phase containing 6% methanol in the same aqueous buffer plus counterion was used. Extraction of these cyclic nucleotides from deproteinized biological samples using disposable reverse-phase extraction columns is described. This not only eliminated lipophilic contaminants, but also served to concentrate the samples. The outlined procedures were used to determine the concentrations of the butyryl derivatives in lung tissue and perfusate following a 35-min lung perfusion with 100 microM N6-O2'-dibutyryl cyclic AMP. The role of this technique in the analysis of cyclic nucleotide derivatives as compared with conventional assay procedures is discussed.