sodium-dodecyl-sulfate and Kidney-Diseases

The goal of replacement, refinement and reduction of animal testing is critically dependent on the development and assessment of novel in vitro methodologies and the further development of existing methodologies. Here, we evaluated the use of a modified perfusion cell culture apparatus for application to chronic in vitro nephrotoxicity testing using DMSO, SDS, paracetamol and cyclosporine A as test compounds. Renal epithelial monolayers were cultured on microporous growth supports and exposed to test compounds under static or perfusion conditions. Alamar Blue reduction, gamma-glutamyl transpeptidase activity (GGT), lactate dehydrogenase activity (LDH) and remnant protein were used to assay cell toxicity. There was no significant difference in IC(50) values between static and perfusion cultures up to 72 hours exposure. However, the perfusion system allowed continuous real-time monitoring of plasma membrane damage, which gives important information of time, duration and scale of toxicity. The complexity of the system restrains its use to low-throughput analysis. However, the real and theoretical advantages of this and similar systems merit further investigations.

Topics: Acetaminophen; Analgesics, Non-Narcotic; Animal Testing Alternatives; Animals; Cell Culture Techniques; Coloring Agents; Cyclosporine; Dimethyl Sulfoxide; Dose-Response Relationship, Drug; gamma-Glutamyltransferase; Humans; Immunosuppressive Agents; Inhibitory Concentration 50; Kidney Diseases; L-Lactate Dehydrogenase; LLC-PK1 Cells; Oxazines; Perfusion; Sodium Dodecyl Sulfate; Surface-Active Agents; Swine; Time Factors; Toxicity Tests, Chronic; Xanthenes

To evaluate results of SDS-agarose gel electrophoresis (AGE) of urinary proteins for use in defining glomerular and tubulointerstitial derangements, investigate patterns of high-molecular-weight (HMW) proteins for differentiating among glomerular disorders, and assess low-molecular-weight (LMW) proteins as markers of severity of tubulointerstitial disease in dogs.. 49 dogs with increased serum creatinine concentrations or abnormal renal protein loss.. Urinary proteins were examined by use of SDS-AGE and differentiated on the basis of molecular weight. The HMW proteins (> or = 69 kd) were considered indicative of glomerular origin, whereas LMW proteins (< 69 kd) were of tubular origin. Renal specimens were examined by use of light microscopy. Glomerular and tubulointerstitial lesions were differentiated by use of the classification for the World Health Organization and semiquantitative grading, respectively.. Sensitivity of SDS-AGE was 100% for detection of glomerular lesions and 92.6% for tubulointerstitial lesions; specificity was 40% and 62.5%, respectively. Although HMW urinary proteins were not significantly associated with the type of glomerular lesion, LMW urinary proteins were significantly associated with the grade of tubulointerstitial damage. Detection of 12- or 15-kd proteins or both was highly indicative of a severe tubulointerstitial lesion.. SDS-AGE of urinary proteins in dogs represents a noninvasive test with high sensitivity for identifying glomerular and tubulointerstitial damage, but low specificity limits its validity as a stand-alone test to differentiate between glomerular and tubulointerstitial lesions. The test is particularly useful for identifying dogs with advanced tubulointerstitial disease but cannot be used to characterize glomerular disorders.

Topics: Animals; Creatine; Dog Diseases; Dogs; Electrophoresis, Agar Gel; Kidney Diseases; Kidney Glomerulus; Proteinuria; Sensitivity and Specificity; Sodium Dodecyl Sulfate

This paper reports the morphological changes in the kidney and spleen of gilthead (Sparus aurata, L), caused by acute action of the anionic detergent, sodium dodecyl sulphate (SDS). Twenty-five giltheads were exposed to SDS concentrations of 5, 8.5, 10 and 15 mg/l. Morphological changes depending on detergent concentrations and length of exposure were seen. Kidney shows loss of normal structure with tubular and renal corpuscle retraction; spleen shows tendency to damage the reticulae structure and a progressive increase of leucocytes and red cells infiltration.

Topics: Animals; Coloring Agents; Kidney; Kidney Cortex; Kidney Diseases; Perciformes; Sodium Dodecyl Sulfate; Spleen; Splenic Diseases

The guinea pigs were dermally exposed to nickel (Ni), sodium lauryl sulphate (SLS) and in their combination for 7 and 14 days. The exposure to Ni and SLS produced changes in enzymes and lipid peroxidation in kidney. The exposure to Ni or SLS depicted slight changes while combined exposure to Ni plus SLS exhibited more degenerative changes in kidney. The result of the study suggests that industrial workers and/or populations exposed simultaneously to Ni and SLS produces more damage to kidney.

Topics: Administration, Cutaneous; Animals; Guinea Pigs; Kidney Diseases; Lipid Peroxidation; Nickel; Skin Absorption; Sodium Dodecyl Sulfate

Renal toxicity is a common manifestation to the exposure of laboratory animals and humans to a wide range of xenobiotics. Traditional methods for evaluating renal damage by clinical chemistry such as blood urea nitrogen (BUN) and serum creatinine are not sensitive to early, mild changes. The use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to measure the molecular weight spectrum of urinary proteins allows for an evaluation of the functional changes associated with renal damage. The ability of the kidney to filter and reabsorb proteins is related to the functional ability of glomeruli and the proximal tubules. Gentamicin sulfate produces injury to the S-1 and S-2 segments of the proximal tubule in laboratory animals and humans. While severe damage to the tubules is associated with increased BUN, serum creatinine, and N-acetyl-beta-glucosiminadase (NAG), mild injury is not detected by these means. The evaluation of urinary proteins by SDS-PAGE demonstrated renal toxicity at a dose of 6 mg/kg after 2 days of sc treatment. The NAG: creatinine ratio was shown to be elevated after 2 days of treatment at 63 mg/kg. The use of SDS-PAGE as described in this paper provides a sensitive method for detecting renal injury.

Topics: Acetylglucosaminidase; Animals; Blood Urea Nitrogen; Creatinine; Dose-Response Relationship, Drug; Electrophoresis, Polyacrylamide Gel; Female; Gentamicins; Kidney; Kidney Diseases; Kidney Tubules, Proximal; Male; Molecular Weight; Proteinuria; Rats; Rats, Sprague-Dawley; Sodium Dodecyl Sulfate

Immunoblotting methods have allowed the identification of 14 urinary proteins. From this study, it appears that some low-relative-molecular mass proteins are derived from disrupted high-relative-molecular mass proteins. The only reliable bands for the diagnosis of tubular lesions, on the polyacrylamide gel stained with Coomassie Blue, are retinol-binding-protein and beta 2-microglobulin. alpha 1-microglobulin can overlap with breakdown products of albumin. The visualization of Ig light chains and lysozyme is rather poor after Coomassie Blue stainings and the accurate identification of these bands may be only improved by the use of immunoblotting.

Topics: Collodion; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Humans; Immunochemistry; Immunoglobulin G; Kidney Diseases; Kidney Tubules; Orosomucoid; Proteinuria; Sodium Dodecyl Sulfate

Topics: Adolescent; Adult; Aged; Child; Diabetes Mellitus; Electrophoresis, Polyacrylamide Gel; Female; Humans; Hypertension; Kidney Diseases; Kidney Glomerulus; Kidney Tubules; Male; Methods; Middle Aged; Molecular Weight; Nephrotic Syndrome; Proteinuria; Sodium Dodecyl Sulfate; Urine

Topics: Animals; Cellulose; Chromatography, Gel; Diabetes Mellitus; Dialysis; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Evaluation Studies as Topic; Horses; Humans; Hydrogen-Ion Concentration; Hypertension; Immunoelectrophoresis; Kidney Diseases; Methods; Molecular Weight; Proteinuria; Sodium Dodecyl Sulfate; Time Factors; Ultrafiltration

Topics: Albuminuria; Bence Jones Protein; Blood Proteins; Chromatography, Gel; Electrophoresis, Disc; Fructose-Bisphosphate Aldolase; gamma-Globulins; Humans; Kidney Diseases; Kidney Glomerulus; Kidney Tubules; L-Lactate Dehydrogenase; Methods; Molecular Weight; Muramidase; Ovalbumin; Pepsin A; Protein Binding; Proteinuria; Sodium Dodecyl Sulfate

Topics: Acute Kidney Injury; Blood Proteins; Creatinine; Electrophoresis, Polyacrylamide Gel; Fanconi Syndrome; Glomerular Filtration Rate; Humans; Immunoglobulin Fragments; Immunoglobulin G; Kidney; Kidney Diseases; Kidney Glomerulus; Nephrons; Nephrotic Syndrome; Proteinuria; Renal Tubular Transport, Inborn Errors; Sodium Dodecyl Sulfate; Uremia