SUMMARY. A new, totally enzymatic procedure for the determination of creatinine in serum and urine, using creatinine amidohydrolase, creatine amidinohydrolase, sarcosine oxidase and formaldehyde dehydrogenase is described. The assay was adapted to a discontinuous analyser with each analysis requiring only 20 J.lL of serum or 3 J.lL of urine. Analytical recovery of creatinine in serum and urine averaged 100· 61170. Within-run and between-run precision studies gave coefficients of variation of 1·1 % and I· 8%, respectively, for a serum with mean values of 83 J.lmollL (9' 4 mg/L) creatinine.Creatinine concentrations in serum and urine were measured by this procedure, in Japanese children and adults. The reference intervals for serum creatinine concentrations in adults 'were 55-96J.lmol/L (6'2-10'9mg/L) in men and 40-66J.lmollL (4'5-7'5mg/L) in women, and for urine, 9'46-19'0Immollday (1070-2150 mg/day) in men and 6,75-10,61 mmollday (764-1200 mg/day) in women. The reference intervals of creatinine clearance were 88· 0-176' 4 mL/min in men and 75'7-173'OmL/min in women.
Additional key phrases: creatine and creatinine amidohydrolases; interference with enzymatic methodUsing our procedure, creatine is produced from creatinine by creatinine amidohydrolase (EC 3.5.2.10) and degraded to sarcosine and urea by creatine amidinohydrolase (BC 3.5.3.3). The sarcosine formed then reacts with sarcosine oxidase (BC 1.5.3.1), in the presence of formaldehyde dehydrogenase (BC 1.2.1.1) and NAD. The NADH + H+ produced is measured at 340 nm.
Splenectomy and TNF-α inhibition both protect the kidney from I/R injury by reducing the accumulation of renal macrophages/monocytes and induction of major inflammatory cytokines.
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