Quantitation of α2u-Globulin in Rat Kidney Cytosol by Capillary Electrophoresis

1999 ◽  
Vol 267 (1) ◽  
pp. 203-211 ◽  
Author(s):  
Axel Pähler ◽  
Kai Blumbach ◽  
Jörg Herbst ◽  
Wolfgang Dekant
Keyword(s):  
Capillary Electrophoresis ◽  
Rat Kidney ◽  
Rat Kidney Cytosol

Fate and Toxicity of 2-(Fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene (Compound A)-derived Mercapturates in Male, Fischer 344 Rats 

1998 ◽  
Vol 89 (5) ◽  
pp. 1174-1183 ◽  
Author(s):  
Vinita Uttamsingh ◽  
Iyer A. Ramswamy ◽  
Raymond B. Baggs ◽  
M. W. Anders
Keyword(s):  
Spectroscopic Analysis ◽  
Rat Kidney ◽  
Urinary Metabolites ◽  
Fischer 344 Rats ◽  
Compound A ◽  
Fischer 344 ◽  
Mercapturic Acid Pathway ◽  
Magnetic Resonance Spectroscopic Analysis ◽  
Acid Pathway ◽  
Rat Kidney Cytosol

Background 2-(Fluoromethoxy)-1,1,3,3,3-pentafluoro-1-propene (compound A) is formed in the anesthesia circuit by the degradation of sevoflurane. Compound A is nephrotoxic in rats and undergoes metabolism by the mercapturic acid pathway in rats and humans to yield the mercapturates S-[2-(fluoromethoxy)-1,1,3,3,3-pentafluoropropyl]-N-acetyl-L -cysteine (compound 3) and S-[2(fluoromethoxy)-1,3,3,3-tetrafluoro-1-propenyl]-N-acetyl-L-cys teine (compound 5). These experiments were designed to examine the fate and nephrotoxicity of compound A-derived mercapturates in rats. Methods The deacetylation of compounds 3 and 5 by human and rat kidney cytosol and with purified acylases I and III was measured, and their nephrotoxicity was studied in male Fischer 344 rats. The metabolism of the deuterated analogs of compounds 3 and 5, [acetyl-2H3]S-[2-(fluoromethoxy)-1,1,3,3,3-pentafluoropropyl ]-N-acetyl-L-cysteine (compound 3-d3) and [acetyl-2H3]S-[2-(fluoromethoxy)-1,3,3,3-tetrafluoro-1-propenyl]-N -acetyl-L-cysteine (compound 5-d3), respectively, was measured. Results Compound 5, but not compound 3, was hydrolyzed by human and rat kidney cytosols and by acylases I and III. 19F nuclear magnetic resonance spectroscopic analysis showed no urinary metabolites of compound 3, but unchanged compound 5 and its metabolites 2-(fluoromethoxy)-3,3,3-trifluoropropanoic acid and 2-[1-(fluoromethoxy)-2,2,2-trifluoroethyl]-4,5-dihydro-1,3-thiazol e-4-carboxylic acid were detected in urine. Compound 5 (250 microM/kg) produced clinical chemical and morphologic evidence of renal injury in two of three animals studied. Conclusions Compounds 3 and 5 underwent little metabolism. Compound 5, but not compound 3, was mildly nephrotoxic. These results indicate that compound A-derived mercapturate formation constitutes a detoxication pathway for compound A.

scholarly journals Characterization of glutathione S-transferases in rat kidney. Alteration of composition by cis-platinum

1988 ◽  
Vol 252 (1) ◽  
pp. 127-136 ◽  
Author(s):  
G M Trakshel ◽  
M D Maines
Keyword(s):  
Specific Activity ◽  
Rat Kidney ◽  
Male Rats ◽  
Glutathione S Transferase ◽  
Aberrant Form ◽  
Glutathione S Transferases ◽  
Exchange Matrix ◽  
Rat Kidney Cytosol

We have developed chromatographic and mathematical protocols that allowed the high resolution of glutathione S-transferase (GST) subunits, and the identification of a previously unresolved GST monomer in rat kidney cytosol; the monomer was identified tentatively as subunit 6. Also, an aberrant form of GST 7-7 dimer appeared to be present in the kidney. This development was utilized to illustrate the response of rat kidney GST following cis-platinum treatment in vivo. Rat kidney cytosol was separated into three ‘affinity families’ of GST activity after elution from a GSH-agarose matrix. The affinity peaks were characterized by quantitative differences in their subunit and dimeric compositions as determined by subsequent chromatography on a cation-exchange matrix and specific activity towards substrates. By use of these criteria, the major GST dimers of affinity peaks were tentatively identified. The major GST dimers in peak I were GST 1-1 and 1-2, in affinity peak II it was GST 2-2, and in peak III they were GST 3-3 and 7-7. GST 3-6 and/or 4-6, which have not been previously resolved in kidney cytosol, were also present in peak II. Alterations in the kidney cytosolic GST composition of male rats were detected subsequent to the administration of cis-platinum (7.0 mg/kg subcutaneously, 6 days). This treatment caused a pronounced alteration in the GST profile, and the pattern of alteration was markedly different from that reported for other chemicals in the kidney or in the liver. In general, the cellular contents of the GSTs of the Alpha and the Mu classes decreased and increased respectively. It is postulated that the decrease in the Alpha class of GSTs by cis-platinum treatment may be related to renal cortical damage and the loss of GSTs in the urine. The increase in the Mu class of GSTs could potentially stem from a lowered serum concentration of testosterone; the latter is a known effect of cis-platinum treatment.

scholarly journals Regulation of aldosterone receptor in rat kidney cytosol by atrial natriuretic factor.

Hypertension ◽  
1989 ◽  
Vol 13 (4) ◽  
pp. 334-340 ◽  
Author(s):  
M Horiuchi ◽  
N Kohashi ◽  
H Nishiyama ◽  
J Hama ◽  
T Takenaka ◽  
...  
Keyword(s):  
Atrial Natriuretic Factor ◽  
Rat Kidney ◽  
Natriuretic Factor ◽  
Aldosterone Receptor ◽  
Rat Kidney Cytosol ◽  
Atrial Natriuretic

scholarly journals Isoenzymes of glutathione transferase in rat kidney cytosol

1985 ◽  
Vol 230 (3) ◽  
pp. 609-615 ◽  
Author(s):  
C Guthenberg ◽  
H Jensson ◽  
L Nyström ◽  
E Österlund ◽  
M K Tahir ◽  
...  
Keyword(s):  
Rat Liver ◽  
Liver Enzymes ◽  
Glutathione Transferase ◽  
Ethacrynic Acid ◽  
Rat Kidney ◽  
Glutathione Transferases ◽  
Substrate Specificities ◽  
Additional Peak ◽  
Rat Kidney Cytosol ◽  
Rat Liver Cytosol

Glutathione transferases from rat kidney cytosol were purified about 40-fold by chromatography on S-hexylglutathione linked to epoxy-activated Sepharose 6B. Further purification by fast protein liquid chromatography with chromatofocusing in the pH interval 10.6-7.6 resolved five major peaks of activity with 1-chloro-2,4-dinitrobenzene as the second substrate. Four of the peaks were identified with rat liver transferases 1-1, 1-2, 2-2 and 4-4 respectively. The criteria used for identification included physical properties, reactions with specific antibodies, substrate specificities and sensitivities to several inhibitors. The fourth major peak is a ‘new’ form of transferase, which has not been found in rat liver. This isoenzyme, glutathione transferase 7-7, has a lower apparent subunit Mr than any of the transferases isolated from rat liver cytosol, and does not react with antibodies raised against the liver enzymes. Glutathione transferases 3-3 and 3-4, which are abundant in liver, were only present in very small amounts. In a separate chromatofocusing separation in a lower pH interval, an additional peak was eluted at pH 6.3. This isoenzyme is characterized by its high activity with ethacrynic acid.

Some problems associated with assay of 25-hydroxycalciferol in human serum.

1977 ◽  
Vol 23 (5) ◽  
pp. 806-810 ◽  
Author(s):  
R S Mason ◽  
S Posen
Keyword(s):  
Human Serum ◽  
Silicic Acid ◽  
Rat Kidney ◽  
Extraction Procedure ◽  
Normal Subjects ◽  
Good Alternative ◽  
Alternative Methods ◽  
Control Serum ◽  
Free Material ◽  
Rat Kidney Cytosol

Abstract Methods available for the assay of 25-hydroxycalciferol in human serum are evaluated and compared to one another. Ethanol was chosen for use in the initial extraction procedure and rat-kidney cytosol as the binding protein, although good alternative methods are also available. We used silicic acid for chromatography and found this an essential step. Reproducibility was increased when, after "bound" and "free" material were separated, an aliquot of the supernate was pipetted into the counting vial instead of the entire supernatant fluid being decanted. Beta-lipo-protein added to the assay system was of no advantage; added bovine serum albumin interfered with the assay by giving rise to high blank values. With ethanol extraction, silicic acid chromatography, rat kidney cytosol and separation on dextran-coated charcoal, sera from normal subjects showed a mean 25-hydroxycalciferol concentration of 28.5 microng/liter (range, 13.1 to 43.9) during the fall season. Coefficients of variation for a control serum were 4.9% (intra-assay) and 10.9% (interassay).

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