The aim of this study is to define the role of human C-reactive protein (CRP) in lipid metabolism, especially in relation to the binding capacity of CRP with serum lipoproteins (LP) by using gel filtration. The uptake of the CRP-LP complex through macrophage was observed by means of a light microscope.
Column chromatography of hyperlipidemic human serum with or without the addition of CRP revealed that a complex was formed between CRP and LP, when the serum contained more than 180mg/dl of total cholesterol and also with more than 110mg/dl of triglyceride. CRP binds sufficiently with LP if the column is passed through with serum in which the titer of total cholesterol is around 250-299mg/dl. Very low density lipoproteins (VLDL) were found to bind with CRP in the hyperlipidemic serum. Binding of CRP with LP was calcium ion-dependent and was inhibited by ligands such as phosphorylcholine or 6-amino-n-caproic acid. The CRP-LP complex was ingested by macrophage.

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Rates of oxidation of bilirubin species were determined spectrometrically using enzymatic oxidation of bilirubin by peroxidase (POD: EC 1.11.1.7) and hydrogen peroxide. Rates of oxidation of conjugated and unconjugated bilirubin (Bc and Bu) were examined with and without serum albumin and compared, and those of bilirubin bound covalently to albumin were determined using naturally occurring delta-bilirubin (Bd). In the presence of serum albumin (Alb), Bc-Alb showed the highest rate of oxidation by POD oxidation followed by Bu-Alb and Bd. Without serum albumin Bu showed a higher rate of oxidation than Bc, despite the fact that Bc exerted more interference than Bu on the enzymatic determination of uric acid (coupled with POD reaction and dye formation).

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We describe a new enzymatic assay method to measure sulfated bile acids in urine, in. which the essential components are a novel bile acid sulfate sulfatase (BSS) and β-hydroxysteroid dehydrogenase (β-HSD: EC 1.1.1.1.51). The BSS produced inducibly by Pseudomonas testosteroni was able to efficiently hydrolyze various kinds of bile acid 3α-sulfates to form 3β-hydroxyl bile acids and sulfuric acid. Total sulfated bile acid (TSBA) in urine was determined by the combined reaction of desulfation of bile acid 3α-sulfates by BSS, followed by dehydrogenation of desulfated substrates by β-HSD and conventional colorimetric assay of reduced nicotinamide adenine dinucleotide (NADH) as formazan dye in the presence of nitrotetrazolium blue and diaphorase. The calibration curve for glycolithocholic acid 3-sulfate as the standard was linear up to 250μmol/l. Analytical recovery of various sulfated bile acids in urine averaged 98% excepting the recovery of taurolithocholic acid 3-sulfate which was approximately 80%. The CV for intra-and inter-assay variations was ≤3.2% and ≤3.4%, respectively. This method is accurate and simple, and less time-consuming than those previously reported. We determined the concentration of TSBA in adult urine samples by this method, and observed that urinary TSBA of normal subjects was less than 7.6μmol/g creatinine (mean2.0±1.8μmol/g creatinine), while in patients with acute hepatitis, liver cirrhosis, and intra-and extra-hepatic biliary obstruction urinary TSBA was markedly increased.

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We describe a simple, sensitive and rapid counting immunoassay (CIA) method which we use for measuring serum C-reactive protein (CRP) with a PAMIA-20 (Sysmex), based on latex agglutination and particle counting. The detection limit was 0.1μg/dl which is equivalent to that of RIA. The within-assay coefficient of variation (CV) ranged from 0.96% to 2.59% in the concentration range of 8.05-152.30μg/dl. The between-assay CV ranged from 2.51% to 3.41% in the concentration range of 16.2-8553.8μg/dl. The results of the counting immunoassay correlated well with those of latex photometric immunoassay using LPIA-300 (n=176, r= 0.997, y=0.969x+17.14). There was no difference in CRP values between serum and EDTA-plasma. The results obtained for the serum CRP of 234 healthy adults (119 male and 115 female) ranging from twenty to sixty years old showed the log-normal distribution in the range of 3.03-159.25μg/dl and a median value of 21.96μg/dl. Male subjects exhibited higher values (median; 29.30μg/dl) of serum CRP than those of female (median; 16.49μg/dl). Plasma CRP of 130 neonates displayed a great change several days after birth. In non-infected infants, the average plasma CRP was 119.1μg/dl (n=10, SD=108.8) on the first day. It showed a peak concentration of 432.8μg/dl (n=41, SD=536.5) on the second day, then decreased exponentially and finally reached a plateau level of 11.8μg/dl (n=7, SD=6.6) on the 7th day.

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1. In a comparison between modified ALBK (Abell-Levy-Brodie-Kendall) method and the enzymatic method of serum total cholesterol, the enzymatic method had better precision from the results of simultaneous multiplex measurement, duplicate random measurements and differences of results among institutions.
2. The measurement of total cholesterol in SRM 909, that is freeze-dried serum sample, by modified ALBK method and the enzymatic method led to low accuracy in the enzymatic method. While the measurement of total cholesterol in fresh serum led to the same level accuracy between these methods.
3. From these findings, it was considered that the enzymatic method may be used for the standardization of serum total cholesterol measurement, if the intact (no freeze-dried) standard serum is prepared.

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Problems exist for the improvement of the latex agglutination method for fecal occult blood testing. The main problems involve the necessity for the punctuality of reaction time and the need for skills in the judgment of test result.
With the intention of improving on these problems, we developed an immuno-gold staining method for fecal occult blood testing using colloidal gold bound with and human hemoglobin monoclonal antibody and membrane bound with anti human hemoglobin.
When the 0.2-400μg/ml of human hemoglobin were tested by this method, the clear purple color of colloidal gold appeared on the membrane. The reaction was completed within 3min and the result color did not change for 60min.
As far as testing was concerned, the other animal hemoglobins, the food materials and the drugs had no effect on the test result by this method. There was a good correlation between the test results of the 499 samples obtained at hospitals by this method, by the latex agglutination method and by the hemagglutination method.
In the three methods, the test results obtained by this method was correlated very well with the results of endoscope testing. As mentioned above, this method is very well suited for operation and may be used as a routine testing fecal occult blood.

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