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Renal Effects
Published in Lars Friberg, Tord Kjellström, Carl-Gustaf Elinder, Gunnar F. Nordberg, Cadmium and Health: A Toxicological and Epidemiological Appraisal, 2019
Piscator193 showed that neither TCA nor SA totally precipitates all the protein in urine. The best precipitation was achieved by the “Tsuchiya” phosphotungstic acid reagent. This was confirmed by Shiroishi and co-workers226 who also showed that the sulfosalicylic acid precipitation is insufficient when the total urinary protein concentration is below 250 mg/ℓ.
Tissue Glutathione
Published in Robert A. Greenwald, CRC Handbook of Methods for Oxygen Radical Research, 2018
Picric, 5-sulfosalicylic, metaphosphoric, trichloroacetic, and perchloric acids have all been used for tissue homogenization and deproteinization. Picric acid (1% w/v) or 5-sulfosalicylic acid (SSA; 5% w/v) are preferred for homogenization and deproteinization because perchloric, trichloroacetic, and metaphosphoric acids may not maintain the GSH to GSSG ratio in all tissues.30
Community-Based Approach to Prevention of Chronic Kidney Disease: The Chennai Experience
Published in Meguid El Nahas, Kidney Diseases in the Developing World and Ethnic Minorities, 2005
In a pilot project before the start of the main program, urine dipsticks had been found to be unreliable. A number of subjects reported to have albuminuria when tested in the field were found to be quite normal when the test was repeated in the hospital. It is possible that transport and storage of the strips under the hot conditions and open sunlight in the field rendered them less reliable. Therefore, the standard and time-tested methods of sulfosalicylic acid for protein and Benedict’s reagent for glucose were used and the results have been found to be reproducible when the patients are brought to the hospital for detailed investigation.
Bromosulfophthalein suppresses inflammatory effects in lipopolysaccharide-stimulated RAW264.7 macrophages
Published in Immunopharmacology and Immunotoxicology, 2020
Fuai Cui, Sean B. Sequeira, Zhenyue Huang, Guowei Shang, Quanjun Cui, Xinlin Yang
The amounts of GSH, nitrite, and TNFα for cells growing on a 96-well plate were measured by the GSH assay kit (Sigma-Aldrich, St. Louis, MO), the Griess reagent (Promega, Madison, WI), and the ELISA kit (Sigma-Aldrich, St. Louis, MO), respectively, following the instructions provided by the manufacturers. The culture medium was first collected for measuring nitrite and TNFα production. To prepare samples for intracellular GSH level, 50 μL of 5% 5-sulfosalicylic acid solution was added to each well after removal of the culture medium. The cells were then frozen under −80 °C and thawed at room temperature (RT) and left at 4 °C for 5 min. The cell lysis was centrifuged at 10,000×g for 10 min. The supernatants were collected and GSH values were determined by a specific color reaction as instructed. All GSH, nitrite and TNFα were normalized by cell number obtained from the MTS assay. The autofluorescence of NAD(P)H, including hydrated nicotinamide adenine dinucleotide (NADH) and nicotinamide adenine dinucleotide phosphate (NADPH), was determined on a BD LSRFortessa™ cell analyzer (San Jose, CA) with UV (351–365 nm) for excitation and a 450/50 nm BP filter for emission detection [11]. The same cell analyzer was also used for assessing expression of CD11b and Fas using an FITC-labeled rat anti-mouse CD11b (clone: M1/70) and hamster anti-mouse Fas (clone: JO2) antibodies (Fisher Scientific, Hampton, NH), respectively. A total of 10,000 cells were harvested per sample, and FCS Express 6 Plus software was used for data analysis.
Impact of acute and subchronic inhalation exposure to PbO nanoparticles on mice
Published in Nanotoxicology, 2018
J. Lebedová, Z. Nováková, Z. Večeřa, M. Buchtová, J. Dumková, B. Dočekal, L. Bláhová, P. Mikuška, I. Míšek, A. Hampl, K. Hilscherová
The measurement of LPO was performed by HPLC/DAD analysis of thiobarbituric acid reactive substances (TBARS). The method is based on measuring of colored substances resulting from reaction of lipid peroxidation products and thiobarbituric acid (Lushchak et al. 2005; Bastos et al. 2012). Degree of lipid peroxidation is expressed as nmol TBARS per miligram of protein. GST activity was measured spectrofotometrically at 340 nm using 1-chloro-2,4-dinitrobenzene and GSH (Habig, Pabst, and Jakoby 1974). Specific activity was expressed as nmol of the product per minute per milligram of protein. Protein concentrations were determined by the method using the Folin-Ciocalteu phenol reagent (Lowry et al. 1951). GSH and GSSG were analyzed by LC–MS/MS. Samples were mixed with internal standard solution of glutathione glycine 13C2, 15N and treated with DTNB. The reaction was stopped by the addition of sulfosalicylic acid. Second internal standard (glutathione disulfide 13C4, 15N2) was injected directly into the samples before analyzes. Concentrations of GSH and GSSG were determined by correcting their responses in mass detector for the response of internal standards. Results were reported as nmol GSH or GSSG per gram of wet tissue.
Clinical spectrum of immunoglobulin A vasculitis in children and determining the best timing of urine examination to predict renal involvement
Published in Postgraduate Medicine, 2022
Fatma Yazılıtaş, Evrim Kargın Çakıcı, Eda Didem Kurt Şükür, Semanur Özdel, Tülin Güngör, Esra Bağlan, Evra Çelikkaya, Deniz Karakaya, Diclehan Orhan, Mehmet Bülbül
Proteinuria was first investigated by the sulfosalicylic acid method, which semi-quantitatively showed urinary proteins containing both albumin and low molecular weight proteins. The proteinuria was calculated using the spot urinary protein creatinine ratio and 24-hour urine protein excretion. Proteinuria was defined as the presence of protein in the early morning urine spot protein creatinine ratio greater than 0.2 mg/mg or in 24-hour urine higher than 4 mg/m2/hr. Nephrotic range proteinuria was defined as protein excretion of greater than 40 mg/m2/hr in 24-hr collected urine or spot protein creatinine ratio greater than 2 mg/mg in the early morning urine [7].