A pilot randomized controlled trial of renal protection with pioglitazone in diabetic nephropathy.

BACKGROUND: Diabetic nephropathy progresses relentlessly to end-stage renal disease (ESRD). Animal experiments have found that peroxisome proliferator activated receptor-gamma (PPAR-gamma)-based therapy can have a glucose independent effect on renal protection. We hypothesized that PPAR-gamma-based antidiabetic therapy would result in greater reduction in proteinuria compared to sulfonylurea-based therapy. METHODS: In 44 patients with overt diabetic nephropathy, an open-label, blinded end point trial was conducted in which subjects were randomized to either pioglitazone or glipizide to achieve similar glucose control. Proteinuria was assessed by two collections of 24-hour urine samples each month for 4 months. RESULTS: The glipizide group had an adjusted mean increase in proteinuria of 6.1% (95% CI -11.7%, 23.8%), whereas the pioglitazone group had a reduction of 7.2% (95% CI -24.9%, 10.6%). The adjusted reduction with pioglitazone of 13.2% (95% CI -38.4%, 11.9%) was not statistically significant (P= 0.294). Baseline proteinuria, diastolic ambulatory blood pressure, and serum albumin concentration were independent predictors of reduction in proteinuria. The frequency and patterns of adverse events were similar in the two groups. CONCLUSION: In patients with advanced diabetic nephropathy, we found no reduction in proteinuria over 4 months. These data are useful to design larger studies with longer duration of follow-up to demonstrate renal protection of PPAR-gamma agonists.

Evaluation of kidney function in patients with acute renal failure using high-performance liquid chromatography: a case report.

Accurate measurement of glomerular function rate (GFR) in the setting of acute renal failure is difficult to achieve with current clinical methods, such as measuring plasma creatinine concentration and 24-hour urine creatinine clearance. High-performance liquid chromatography was used to measure GFR directly in a critically ill patient with acute renal failure. This approach involved evaluating the elimination kinetics of nonionic contrast material administered intravenously for radiologic imaging. It required no additional patient exposure to radiographic contrast media and enabled caregivers to determine kidney function accurately in the presence of worsening clinical status and delayed changes in plasma creatinine. This and other methods for more accurate measurement of GFR in patients with acute renal failure may provide the foundation for clinical studies that assess the severity and management of acute renal failure.

Liquid chromatography for iothalamate in biological samples.

We have previously reported an iothalamate assay for the assessment of the glomerular filtration rate (GFR) that required a long column equilibration time and 22 min run time per sample. We now report a simpler assay that requires a run time of only 5.5 min and is more precise and accurate than the earlier technique. The mobile phase consisted of methanol-acetonitrile-50 mM sodium monobasic phosphate (10:5:85, v/v) at pH 4.4, pumped at a rate of 1.5 ml/min on a C(18) reversed-phase column. Samples of plasma and urine were deproteinized with 1 volume of 4% perchloric acid or 9 volumes of 2% perchloric acid, respectively. No internal standard was used. The diode array detection system collected absorbance at 240 nm and the peak height areas of iothalamate were determined. The iothalamate peak appeared at 3.5 min. Detector response was linear over the range tested (10-2000 microg/ml). Within-run precision was <3% for both plasma and urine and accuracy was 96-102%. Between-day precision for plasma and urine analyses were <7%. The recovery of iothalamate in urine and plasma were 102% and 91%, respectively. There was excellent thermal and pH stability of iothalamate. No interference was found with para-amino hippuric acid (PAH) or N-acetyl PAH, which can be simultaneously assayed, if desired.

Rapid, fluorimetric-liquid chromatographic determination of malondialdehyde in biological samples.

Current chromatographic methods of estimation of malondialdehyde, a marker of oxidative lipid injury, often require extensive extraction procedures, column cleaning or specialized equipment. A rapid and sensitive HPLC method is described for the determination of MDA in plasma and urine. The mobile phase consisted of 40:60 ratio (v/v) of methanol to 50 mM potassium monobasic phosphate at pH 6.8, pumped at a rate of 1.0 ml/min on a Hewlett-Packard Hypersil 5 micro ODS 100 x 4.6 mm placed in a column warmer set to 37 degrees C. Samples of plasma and urine were treated with the antioxidant, butylated hydroxytoluene and heat derivatized at 100 degrees C for 1 h with thiobarbituric acid at an acid pH. Samples were extracted with n-butanol and 10 microl of the extract was injected at 1 min intervals using an autosampler. The Hewlett-Packard model 1046A programmable fluorescence detector was set at excitation of 515 nm and emission of 553 nm. Retention time was 1.87 min, however absence of interfering peaks, allowed analysis to be carried out in increments of 1 min per sample. Within day variability in estimation was between 8.6% and 10.3%. Between days variability was 3.6-7.9%. Recovery was between 88 and 101%. Samples of urine and plasma from ten normotensive volunteers were 1.94 +/- 0.79 micromol/g creatinine and 0.69 +/- 0.13 micromol/l respectively and were similar to those reported in the literature. MDA degrades at room temperature at a rate of 10% per hour. It is therefore, suggested that the total assay time be limited to 1 h beginning with sample preparation.