We investigated contents and classes of urinary and stone matrix lipids, and evaluated their clin... more We investigated contents and classes of urinary and stone matrix lipids, and evaluated their clinical relevance in nephrolithiasis patients. Lithogenic role of major lipid classes was explored. Urine (24 h) and stone samples were collected from 47 patients with nephrolithiasis. Control urines were obtained from 29 healthy subjects. Urinary 8-hydroxy-deoxyguanosine (8-OHdG), malondialdehyde (MDA), N-acetyl-β-glucosaminidase (NAG) activity and total proteins were measured. Total lipids were extracted from centrifuged urines (10,000 rpm, 30 min) and stones by chloroform/methanol method. Major classes of lipids were identified using multi-one-dimensional thin-layer chromatography (MOD-TLC). Influence of each lipid class purified from stone matrices on stone formation was evaluated using crystallization and crystal aggregation assays. Urinary NAG activity and 8-OHdG were significantly elevated in nephrolithiasis patients. Total lipids in centrifuged urines of the patients were not significantly different from that of controls. In nephrolithiasis, urinary excretion of total lipids was linearly correlated to urinary MDA, 8-OHdG, NAG activity and total proteins. Lipid contents in stone matrices varied among stone types. Uric acid stone contained lower amount of total lipids than calcium oxalate and magnesium ammonium phosphate stones. MOD-TLC lipid chromatograms of healthy urines, nephrolithiasis urines and stone matrices were obviously different. Triacylglyceride was abundant in urines, but scarcely found in stone matrices. Stone matrices were rich in glycolipids and high-polar lipids (phospholipids/gangliosides). Partially purified glycolipids significantly induced crystal aggregation while cholesterol was a significant inducer of both crystal formation and agglomeration. In conclusion, total lipids in centrifuged urines did not differ between nephrolithiasis and healthy subjects. Our finding suggests that the significant sources of lipids in patients’ urine may be large lipids-containing particles, which are removed in centrifuged urines. However, urinary lipid excretion in nephrolithiasis patients was associated with the extent of oxidative stress and renal tubular injury. Triacylglyceride was abundant in urines, but rarely incorporated into stones. Glycolipids were principal lipid constituents in stone matrices and functioned as crystal aggregator. Cholesterol purified from stone matrices bared crystal nucleating and aggregating activities.
We have previously used agarose gel electrophoresis and immunoblotting to qualitatively measure s... more We have previously used agarose gel electrophoresis and immunoblotting to qualitatively measure serum MUC5AC mucin for diagnosing cholangiocarcinoma. In this study, we developed a quantitative determination of serum MUC5AC by sandwich ELISA using MUC5AC mucin monoclonal antibody and soybean agglutinin. A cut-off value of the absorbance 0.074 was obtained from a complete statistical Receiver Operating Characteristic curves with an area under the curve=0.8141. The assay could discriminate cholangiocarcinoma patients from the controls with 71% sensitivity and 90% specificity. The test is simple to perform, reproducible, and probably used for detecting cholangiocarcinoma in a high-risk group or suspected patients.
8-hydroxydeoxyguanosine (8-OHdG) is an oxidatively modified guanosine, which has been widely used... more 8-hydroxydeoxyguanosine (8-OHdG) is an oxidatively modified guanosine, which has been widely used as an oxidative DNA damage marker in various diseases. The present study aimed to determine urinary 8-OHdG in nephrolithiasis patients and evaluate its clinical significance. Thirty-six nephrolithiasis patients and 30 healthy subjects were recruited. Urine volume, creatinine, malondialdehyde, β-N-acetylglucosaminidase (NAG) activity and proteins were measured in 24 h urine samples. Urinary 8-OHdG was determined by competitive enzyme-linked immunosorbent assay. Mineral composition of stones was analyzed using Fourier-transformed infrared spectroscopy. Nephrolithiasis patients excreted urinary 8-OHdG significantly higher than healthy controls. Urinary 8-OHdG levels compared among patients with calcium oxalate, struvite and uric acid stones were insignificantly different. The urinary NAG activity correlated positively with urinary 8-OHdG. Multiple linear regression showed that urinary NAG activity was an independent predictor of urinary 8-OHdG level. Receiver operating characteristic analysis revealed that the urinary 8-OHdG test was adequate for diagnosing nephrolithiasis. At 10 μg/g creatinine cutoff, the 8-OHdG test imparted high specificity (96.67%) and a positive predictive value (91.67%). In conclusion, this is the first report of elevated urinary 8-OHdG excretion in nephrolithiasis patients indicating increased oxidative DNA damage. Increased renal tubular damage was independently associated with elevated urinary 8-OHdG. Elevated urinary 8-OHdG levels adjunct with metabolic profile may be useful for identifying people at risk of stone development.
Potassium citrate has long been used as a prophylactic remedy for nephrolithiasis recurrence. Lem... more Potassium citrate has long been used as a prophylactic remedy for nephrolithiasis recurrence. Lemonade consumption is also suggested as an option. We compared the efficacy of consumption of solution containing manufactured lime powder with that of potassium citrate, on the improvement of metabolic risk factors, oxidative stress and renal tubular damage in nephrolithiasis patients. Patients with kidney stone were enrolled and randomly assigned to three treatment programs for 3 month period consisting of consumption of solution containing lime powder (Group 1, n = 13), potassium citrate (Group 2, n = 11) and lactose as placebo regimen (Group 3, n = 7). Lime powder and potassium citrate contained equal amounts of potassium (21 mEq) and citrate (63 mEq). After treatment, there was an increase in urinary pH, potassium and citrate in Group 1 and 2. Increased plasma potassium and red blood cell glutathione (R-GSH) and decreased urinary malondialdehyde were found in Group 1, but not observed in Group 2. R-GSH was decreased in Group 2. Urinary N-acetyl-β-glucosaminidase activity and fractional excretion of magnesium, as renal tubular damage indicators, were decreased only in Group 1. In Group 3, all measured parameters were unaltered except for an increased urinary chloride. In conclusion, consumption of our in-house lime powder exerted citraturic and alkalinizing actions as efficient as consumption of potassium citrate. In addition, it provided an antioxidative effect and was able to attenuate renal tubular damage. These pharmacological properties may be clinically useful to diminish the stone-forming potential in kidney stone patients and hence for preventing recurrent calculi.
OBJECTIVESTo investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MC... more OBJECTIVESTo investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in patients with nephrolithiasis, and to evaluate whether their expression is associated with renal function, as oxidative stress and inflammation are involved in the pathogenesis of nephrolithiasis.To investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in patients with nephrolithiasis, and to evaluate whether their expression is associated with renal function, as oxidative stress and inflammation are involved in the pathogenesis of nephrolithiasis.PATIENTS, SUBJECTS AND METHODSRenal biopsies from near the stone, and blood and 24-h urine specimens were collected from 29 patients with nephrolithiasis. Control renal tissues were taken from non-cancerous and cancerous portions of nephrectomy from six patients with renal cancers, and control 24-h urine samples were obtained from 30 healthy subjects. Corrected creatinine clearance, urinary N-acetyl-β-glucosaminidase activity and 8-hydroxy-deoxyguanosine (8-OHdG) were determined. The mRNA expressions of MCP-1 and IL-6 in the tissues were measured by real time reverse transcription-polymerase chain reaction.Renal biopsies from near the stone, and blood and 24-h urine specimens were collected from 29 patients with nephrolithiasis. Control renal tissues were taken from non-cancerous and cancerous portions of nephrectomy from six patients with renal cancers, and control 24-h urine samples were obtained from 30 healthy subjects. Corrected creatinine clearance, urinary N-acetyl-β-glucosaminidase activity and 8-hydroxy-deoxyguanosine (8-OHdG) were determined. The mRNA expressions of MCP-1 and IL-6 in the tissues were measured by real time reverse transcription-polymerase chain reaction.RESULTSPatients with nephrolithiasis had significantly greater renal tubular damage and oxidative stress than the healthy controls. Intrarenal mRNA expressions of MCP-1 and IL-6 in stone-adjacent renal tissues were significantly lower than in cancerous renal tissues, but not statistically different from that in non-cancerous renal tissues. In stone-adjacent renal tissues, the mRNA level of MCP-1 was significantly higher than that of IL-6, but their expressions were significantly correlated with each other. Histological examination showed that the number of infiltrated leukocytes corresponded well with the intrarenal mRNA levels of MCP-1 and IL-6. Patients with nephrolithiasis and compromised renal function had significantly higher intrarenal mRNA levels of MCP-1 and IL-6 than those with preserved renal function. Also, the mRNA levels in patients with severe renal tubular damage were significantly greater than in those with less renal tubular damage. There was no association between intrarenal mRNA expression and urinary 8-OHdG.Patients with nephrolithiasis had significantly greater renal tubular damage and oxidative stress than the healthy controls. Intrarenal mRNA expressions of MCP-1 and IL-6 in stone-adjacent renal tissues were significantly lower than in cancerous renal tissues, but not statistically different from that in non-cancerous renal tissues. In stone-adjacent renal tissues, the mRNA level of MCP-1 was significantly higher than that of IL-6, but their expressions were significantly correlated with each other. Histological examination showed that the number of infiltrated leukocytes corresponded well with the intrarenal mRNA levels of MCP-1 and IL-6. Patients with nephrolithiasis and compromised renal function had significantly higher intrarenal mRNA levels of MCP-1 and IL-6 than those with preserved renal function. Also, the mRNA levels in patients with severe renal tubular damage were significantly greater than in those with less renal tubular damage. There was no association between intrarenal mRNA expression and urinary 8-OHdG.CONCLUSIONNephrolithiasis was associated with low-grade intrarenal inflammation. A greater intrarenal mRNA expression of MCP-1 and IL-6 was associated with enhanced renal impairment. Thus, expression of MCP-1 and IL-6, at least in part, contributed to the progression of nephrolithiasis.Nephrolithiasis was associated with low-grade intrarenal inflammation. A greater intrarenal mRNA expression of MCP-1 and IL-6 was associated with enhanced renal impairment. Thus, expression of MCP-1 and IL-6, at least in part, contributed to the progression of nephrolithiasis.
We evaluated the effects of lifestyle modification (LM) on lipid profile, oxidative stress and se... more We evaluated the effects of lifestyle modification (LM) on lipid profile, oxidative stress and serum-stimulated human coronary artery endothelial cell (HCAEC) viability in coronary artery disease (CAD) patients after 6 months.Thirty patients with CAD were randomly assigned to LM intervention (n = 15) and usual care control (n = 15) groups. LM-intervened patients were instructed to consume low-fat, high-antioxidants and fiber diets. Moderate exercise and stress management were also advised. Group support to maintain patients' compliance was applied.Serum cholesterol, triglyceride, oxidized LDL and protein carbonyl were decreased in LM group. Serum triglyceride was increased in control group. HCAEC viability was increased, while intracellular reactive oxygen species was decreased, by serum from the LM group.LM is capable of improving lipid profile, reducing oxidative stress and increasing HCAEC survival in the patients with CAD, hence lowering a risk for the future cardiovascular event.
We investigated contents and classes of urinary and stone matrix lipids, and evaluated their clin... more We investigated contents and classes of urinary and stone matrix lipids, and evaluated their clinical relevance in nephrolithiasis patients. Lithogenic role of major lipid classes was explored. Urine (24 h) and stone samples were collected from 47 patients with nephrolithiasis. Control urines were obtained from 29 healthy subjects. Urinary 8-hydroxy-deoxyguanosine (8-OHdG), malondialdehyde (MDA), N-acetyl-β-glucosaminidase (NAG) activity and total proteins were measured. Total lipids were extracted from centrifuged urines (10,000 rpm, 30 min) and stones by chloroform/methanol method. Major classes of lipids were identified using multi-one-dimensional thin-layer chromatography (MOD-TLC). Influence of each lipid class purified from stone matrices on stone formation was evaluated using crystallization and crystal aggregation assays. Urinary NAG activity and 8-OHdG were significantly elevated in nephrolithiasis patients. Total lipids in centrifuged urines of the patients were not significantly different from that of controls. In nephrolithiasis, urinary excretion of total lipids was linearly correlated to urinary MDA, 8-OHdG, NAG activity and total proteins. Lipid contents in stone matrices varied among stone types. Uric acid stone contained lower amount of total lipids than calcium oxalate and magnesium ammonium phosphate stones. MOD-TLC lipid chromatograms of healthy urines, nephrolithiasis urines and stone matrices were obviously different. Triacylglyceride was abundant in urines, but scarcely found in stone matrices. Stone matrices were rich in glycolipids and high-polar lipids (phospholipids/gangliosides). Partially purified glycolipids significantly induced crystal aggregation while cholesterol was a significant inducer of both crystal formation and agglomeration. In conclusion, total lipids in centrifuged urines did not differ between nephrolithiasis and healthy subjects. Our finding suggests that the significant sources of lipids in patients’ urine may be large lipids-containing particles, which are removed in centrifuged urines. However, urinary lipid excretion in nephrolithiasis patients was associated with the extent of oxidative stress and renal tubular injury. Triacylglyceride was abundant in urines, but rarely incorporated into stones. Glycolipids were principal lipid constituents in stone matrices and functioned as crystal aggregator. Cholesterol purified from stone matrices bared crystal nucleating and aggregating activities.
We have previously used agarose gel electrophoresis and immunoblotting to qualitatively measure s... more We have previously used agarose gel electrophoresis and immunoblotting to qualitatively measure serum MUC5AC mucin for diagnosing cholangiocarcinoma. In this study, we developed a quantitative determination of serum MUC5AC by sandwich ELISA using MUC5AC mucin monoclonal antibody and soybean agglutinin. A cut-off value of the absorbance 0.074 was obtained from a complete statistical Receiver Operating Characteristic curves with an area under the curve=0.8141. The assay could discriminate cholangiocarcinoma patients from the controls with 71% sensitivity and 90% specificity. The test is simple to perform, reproducible, and probably used for detecting cholangiocarcinoma in a high-risk group or suspected patients.
8-hydroxydeoxyguanosine (8-OHdG) is an oxidatively modified guanosine, which has been widely used... more 8-hydroxydeoxyguanosine (8-OHdG) is an oxidatively modified guanosine, which has been widely used as an oxidative DNA damage marker in various diseases. The present study aimed to determine urinary 8-OHdG in nephrolithiasis patients and evaluate its clinical significance. Thirty-six nephrolithiasis patients and 30 healthy subjects were recruited. Urine volume, creatinine, malondialdehyde, β-N-acetylglucosaminidase (NAG) activity and proteins were measured in 24 h urine samples. Urinary 8-OHdG was determined by competitive enzyme-linked immunosorbent assay. Mineral composition of stones was analyzed using Fourier-transformed infrared spectroscopy. Nephrolithiasis patients excreted urinary 8-OHdG significantly higher than healthy controls. Urinary 8-OHdG levels compared among patients with calcium oxalate, struvite and uric acid stones were insignificantly different. The urinary NAG activity correlated positively with urinary 8-OHdG. Multiple linear regression showed that urinary NAG activity was an independent predictor of urinary 8-OHdG level. Receiver operating characteristic analysis revealed that the urinary 8-OHdG test was adequate for diagnosing nephrolithiasis. At 10 μg/g creatinine cutoff, the 8-OHdG test imparted high specificity (96.67%) and a positive predictive value (91.67%). In conclusion, this is the first report of elevated urinary 8-OHdG excretion in nephrolithiasis patients indicating increased oxidative DNA damage. Increased renal tubular damage was independently associated with elevated urinary 8-OHdG. Elevated urinary 8-OHdG levels adjunct with metabolic profile may be useful for identifying people at risk of stone development.
Potassium citrate has long been used as a prophylactic remedy for nephrolithiasis recurrence. Lem... more Potassium citrate has long been used as a prophylactic remedy for nephrolithiasis recurrence. Lemonade consumption is also suggested as an option. We compared the efficacy of consumption of solution containing manufactured lime powder with that of potassium citrate, on the improvement of metabolic risk factors, oxidative stress and renal tubular damage in nephrolithiasis patients. Patients with kidney stone were enrolled and randomly assigned to three treatment programs for 3 month period consisting of consumption of solution containing lime powder (Group 1, n = 13), potassium citrate (Group 2, n = 11) and lactose as placebo regimen (Group 3, n = 7). Lime powder and potassium citrate contained equal amounts of potassium (21 mEq) and citrate (63 mEq). After treatment, there was an increase in urinary pH, potassium and citrate in Group 1 and 2. Increased plasma potassium and red blood cell glutathione (R-GSH) and decreased urinary malondialdehyde were found in Group 1, but not observed in Group 2. R-GSH was decreased in Group 2. Urinary N-acetyl-β-glucosaminidase activity and fractional excretion of magnesium, as renal tubular damage indicators, were decreased only in Group 1. In Group 3, all measured parameters were unaltered except for an increased urinary chloride. In conclusion, consumption of our in-house lime powder exerted citraturic and alkalinizing actions as efficient as consumption of potassium citrate. In addition, it provided an antioxidative effect and was able to attenuate renal tubular damage. These pharmacological properties may be clinically useful to diminish the stone-forming potential in kidney stone patients and hence for preventing recurrent calculi.
OBJECTIVESTo investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MC... more OBJECTIVESTo investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in patients with nephrolithiasis, and to evaluate whether their expression is associated with renal function, as oxidative stress and inflammation are involved in the pathogenesis of nephrolithiasis.To investigate the intrarenal mRNA expression of monocyte chemoattractant protein-1 (MCP-1) and interleukin-6 (IL-6) in patients with nephrolithiasis, and to evaluate whether their expression is associated with renal function, as oxidative stress and inflammation are involved in the pathogenesis of nephrolithiasis.PATIENTS, SUBJECTS AND METHODSRenal biopsies from near the stone, and blood and 24-h urine specimens were collected from 29 patients with nephrolithiasis. Control renal tissues were taken from non-cancerous and cancerous portions of nephrectomy from six patients with renal cancers, and control 24-h urine samples were obtained from 30 healthy subjects. Corrected creatinine clearance, urinary N-acetyl-β-glucosaminidase activity and 8-hydroxy-deoxyguanosine (8-OHdG) were determined. The mRNA expressions of MCP-1 and IL-6 in the tissues were measured by real time reverse transcription-polymerase chain reaction.Renal biopsies from near the stone, and blood and 24-h urine specimens were collected from 29 patients with nephrolithiasis. Control renal tissues were taken from non-cancerous and cancerous portions of nephrectomy from six patients with renal cancers, and control 24-h urine samples were obtained from 30 healthy subjects. Corrected creatinine clearance, urinary N-acetyl-β-glucosaminidase activity and 8-hydroxy-deoxyguanosine (8-OHdG) were determined. The mRNA expressions of MCP-1 and IL-6 in the tissues were measured by real time reverse transcription-polymerase chain reaction.RESULTSPatients with nephrolithiasis had significantly greater renal tubular damage and oxidative stress than the healthy controls. Intrarenal mRNA expressions of MCP-1 and IL-6 in stone-adjacent renal tissues were significantly lower than in cancerous renal tissues, but not statistically different from that in non-cancerous renal tissues. In stone-adjacent renal tissues, the mRNA level of MCP-1 was significantly higher than that of IL-6, but their expressions were significantly correlated with each other. Histological examination showed that the number of infiltrated leukocytes corresponded well with the intrarenal mRNA levels of MCP-1 and IL-6. Patients with nephrolithiasis and compromised renal function had significantly higher intrarenal mRNA levels of MCP-1 and IL-6 than those with preserved renal function. Also, the mRNA levels in patients with severe renal tubular damage were significantly greater than in those with less renal tubular damage. There was no association between intrarenal mRNA expression and urinary 8-OHdG.Patients with nephrolithiasis had significantly greater renal tubular damage and oxidative stress than the healthy controls. Intrarenal mRNA expressions of MCP-1 and IL-6 in stone-adjacent renal tissues were significantly lower than in cancerous renal tissues, but not statistically different from that in non-cancerous renal tissues. In stone-adjacent renal tissues, the mRNA level of MCP-1 was significantly higher than that of IL-6, but their expressions were significantly correlated with each other. Histological examination showed that the number of infiltrated leukocytes corresponded well with the intrarenal mRNA levels of MCP-1 and IL-6. Patients with nephrolithiasis and compromised renal function had significantly higher intrarenal mRNA levels of MCP-1 and IL-6 than those with preserved renal function. Also, the mRNA levels in patients with severe renal tubular damage were significantly greater than in those with less renal tubular damage. There was no association between intrarenal mRNA expression and urinary 8-OHdG.CONCLUSIONNephrolithiasis was associated with low-grade intrarenal inflammation. A greater intrarenal mRNA expression of MCP-1 and IL-6 was associated with enhanced renal impairment. Thus, expression of MCP-1 and IL-6, at least in part, contributed to the progression of nephrolithiasis.Nephrolithiasis was associated with low-grade intrarenal inflammation. A greater intrarenal mRNA expression of MCP-1 and IL-6 was associated with enhanced renal impairment. Thus, expression of MCP-1 and IL-6, at least in part, contributed to the progression of nephrolithiasis.
We evaluated the effects of lifestyle modification (LM) on lipid profile, oxidative stress and se... more We evaluated the effects of lifestyle modification (LM) on lipid profile, oxidative stress and serum-stimulated human coronary artery endothelial cell (HCAEC) viability in coronary artery disease (CAD) patients after 6 months.Thirty patients with CAD were randomly assigned to LM intervention (n = 15) and usual care control (n = 15) groups. LM-intervened patients were instructed to consume low-fat, high-antioxidants and fiber diets. Moderate exercise and stress management were also advised. Group support to maintain patients' compliance was applied.Serum cholesterol, triglyceride, oxidized LDL and protein carbonyl were decreased in LM group. Serum triglyceride was increased in control group. HCAEC viability was increased, while intracellular reactive oxygen species was decreased, by serum from the LM group.LM is capable of improving lipid profile, reducing oxidative stress and increasing HCAEC survival in the patients with CAD, hence lowering a risk for the future cardiovascular event.
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