Patients with chronic renal failure were divided into two groups according to their hemodialyzed period; the one is a short-term group in which patients had been treated for 2 years, and the other was a long-term group in which they had been treated for 6 years. Their serum cadmium and lead levels were analysed by Zeeman-effect atomic absorption spectrophotometry. Serum cadmium levels in both groups were statistically higher than in the normal healthy group (control group: male 0.36 ppb, female 0.41 ppb; short-term group: male 2.13 ppb, female 1.32 ppb; long-term group: male 3.20ppb, female 2.45 ppb). Al so lead content in long-term group was statistically higher (control group: male 103.75 ppb, femle 102.50 ppb ; short-term group: male 177.50 ppb, female 144.38 ppb; long-term group: male 259.00 ppb, female 215.64 ppb). However, there was no difference in the level of either metal between male and female. These levels in the serum were unchanged at the time of pre- and post-dialysis for single treatment. The same trend was also found in non-protein bound metal levels. The increase of serum cadmium and lead levels in the patients treated with hemodialysis did not result from the dialysis fluid but from exogenous origins such as diet.

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Previonsly, we examined the proteases in normal and metaplastic human stomach mucosa, preparing zymograms of extract fluid from tissue with tosyl-α-lysine-α-naphthylester and α-prolyl-α-phenylalanyl-α-arginene α-naphthylester as substrates. As a result, an additional band (Rf-0.64) newly appeared in both zymograms of the metaplastic mucosa but not in those of normal mucosa. This band has also been observed with zymograms of tissue extracts of normal small intestine and gastric cancer.
In order to confirm the identity of these enzymes (Rf-0.64), which appeared in three kinds of tissue extracts, metaplastic human stomach mocosa, small intestine and gastric cancer, we purified them and examined their properties. The purified enzymes were identified as one and the same from the results of SDS-polyacrylamide gel electrophoresis and zymogram.
The molecular weight of each enzyme was found to be 55,000 and 54,000 by gel filtration on Sephadex G-200 and SDS-PAGE respectively. The effects of various inhibitors on the enzyme activities were almost the same. They were inhibited by DFP, leupeptin, Trasylol and SBTI but not by iodoacetate, EDTA, chymostain and esterastin. Moreover, the result of substrate specificities or optimum pH was almost the same with each enzyme. From these results, it was indicated that the these enzymes were the same and found to be serine protease.
Furthermore, we examined the association between intestinal metaplasia and gastric adenocarcinoma using this enzyme, detected in the preparation of the zymogram, as an indication. Examining two types of gastric cancer, the enzyme was not detected in scirrhus and other organ cancer, but only in well-differentiated adenocarcinom. This indicates that adenocarcinoma may originate from the intestinal metaplastic region.

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A rapid and sensitive single-isotope radioenzymatic assay for norepinephrine has been developed. This method is based on the conversion of norepinephrine to tritiated epinephrine by the enzyme phenylethanolamine-N-methyltransferase (PNMT) and ³H-S-adenosylmethionine (³H-SAM).
100 μl of untreated plasma were added directly to the reaction mixture. After incubation, labelled epinephrine was extracted with alumina and separated from ³H-SAM by precipitation of ³H-SAM with tungstate and adenosine. Adenosine could be substituted for S-adenosylmethionine (SAM) to coprecipitate the remaining radioactive SAM. In order to eliminate the effects of other plasma components, catecholamine-free plasma and an internal standard had to be assayed in parallel with a sample. 400 μl of plasma was sufficient for duplicate determinations of norepinephrine.
This method is simple and 25 plasma samples (100 test tubes) can be processed within 4 hours.

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Decrease of serum triglyceride concentration and reciprocal increase of free fatty acid were observed at 30 minutes after intravenous administration of 300mg dextran sulfate in 5 healthy subjects, while serum total cholesterol and phospholipids were not affected. HDL cholesterol level was tended to increase, which was due to the increase of HDL₂ cholesterol. HDL₃ Cholesterol was not influenced essentially.
Serum VLDL concentration was markedly reduced by dextran sulfate. Although IDL and HDL were elevated, LDL level was unaffected. Above all, IDL triglyceride and HDL lipids incresed concomitantly.
Serum apo A-I, A-II, B, C-II, C-III and E protein concentrations were almost identical when compared with the pretreatment levels. However, apo B, C-II, C-III and E protein concentrations in VLDL fraction decreased remarkably following by the reduction of VLDL concentration. Since the changes of apo C-II and C-III concentration were noticeably, the percentages of these apoproteins in VLDL apoproteins became lower than those in the pretreatment. By contrast, apo C-II, C-III and E proteins increased in HDL fraction. The increases of apo C-II and C-III proteins were more pronaunced than that of apo E. Therefore, those apoproteins are considered to be transferred shift from VLDL to HDL by the lipolysis induced by dextran.
As for serum phospholipid fractions, the percentage of serum lysophosphatidyl choline increased, meanwhile the phosphatidyl choline/lysophosphatidyl choline ratio in serum and the percentage of phosphatidyl ethanolamine in VLDL decreased after the dextran sulfate injection. It suggests that dextran does not only stimulate lipoprotein lipase but also phospholipase activity at the same time as heparin.

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Serum cholinesterase activity were determined by a kinetic method using acetylcholine as substrate (AK method) with a Hitachi 736 autoanalyzer system. Measurement of serum cholinesterase by AK method were linear from 0 to 8000IU/L. The coeffient of variation in within-run was 0.99% for a sample with cholinesterase activity of 3305IU/L, and 0.92% for a sample with activity of 6803IU/L (n=10). The CV in between-run was 1.2% for a sample with activity of 3295IU/L (n=7). Neither ascorbate (up to 50mg/dl) nor bilirubin (up to 40mg/dl) nor hemoglobin (up to 500mg/dl) did interfere with the measurement by this method.
The serum cholinesterase activities of both the human being and several kinds of animals, such as dog, rat, rabbit and monkey, as determined by the pH change (ΔpH) produced by a reaction using acetylcholine as substrate were correlated well with those determined by AK method, but not well as a whole with those measured by a method using o-toluoylcholine as substrate. Some control serum samples show abnormally high activities of cholinesterase when determined by AK method, as compared with the indicated value. Because acetate intereferes with measurement of cholinesterase activity by AK method, these sample sera may be spiked with an acetate solution by the manufacturer without notice.

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Changes in serum manganese superoxide dismutase level in the course of liver diseases were monitored in order to elucidate more precisely the relationship between diseases andserum superoxide dismutase level. Manganese-superoxide dismutase activity had good correlations with some other enzyme activities in patients with acute hepatitis, liver cancer, liver cirrhosis and alcoholic cirrhosis, but it had no correlation with such enzyme activities in case of subacute hepatitis and chronic hepatitis. Studies onthe clinical course in liver diseases demostrated that the change of manganese-superoxide dismutase was later than that of other enzymes. The amounts of manganese-superoxide dismutase may be in- creased not only due to release from the injured cells, but also due to induction in order to exclude the superoxide anion radical which is produced in inflammation.

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Using a radioimmunoassay, we studied the significance of bombesin as a specific marker for small-cell carcinoma of the lung (SCCL). Bombesin-like immunoreactivity (BLI) was detectable in serum4of15patients with SCCL (27%), but was undetectable in serum of all 15 healthy subjects. BLI was also detectable in 7 of 31 patients with non-SCCL tumors (23%) and 4 of 31 patients with other types of lung disease (13%). The present results indicate that bombesin is unlikely to be a specific marker for the early detection of SCCL though bombesin could be formed by SCCL and its metastasis, and might be released into the circulation.

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Antisera against spermine were obtained by immunization of rabbits with o-phthalaldehydic spermine conjugated to bovine serum albumin. Characterization of the antiserum and radioimmunoassay of polyamines were described. Polyamine can be determined after reaction with o-phthalaldehyde. As little as 40 and 30 pmol of respective spermine and spermidine were detected by this assay.

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