Minoru Yakata

Sandwich enzyme immunoassay for serum amyloid A protein (SAA)

Serum amyloid A (SAA), a putative serum precursor of the AA protein which constitutes amyloid fibrils in secondary amyloidosis [l], has been clinically evaluated as one of the sensitive acute-phase reactants in serum [2,3]. The SAA concentration has been analyzed by radial i~unodiffusion (RID) f4,5] and radioi~unoassay (RIA) [6-81. The RID method was very simple and suitable for the analysis of SAA in its increased states, but could not determine the normal range of SAA because of low sensitivity. At present there are only a few descriptions outlining the use of enzyme immunoassay (EIA) in determining the concentration of SAA in serum [8,9]. It has also been mentioned that EIA may be unsuitable for this purpose because of poor reproducibility of results [S]. In this study, we attempted to develop a sensitive and highly reproducible EIA method for the measurement of SAA in serum, and to evaluate the normal range of the SAA.

Determination of urinary cortisol and 6β-hydroxycortisol by high performance liquid chromatography

A method using high performance liquid chromatography for the quantitative determination of cortisol and 6 beta-hydroxycortisol in urine is described. Urine extracts were fractionated by high performance liquid chromatography. The peak of 6 beta-hydroxycortisol was identified on the basis of retention time (24.4 min) and its area was measured. The peak of cortisol could not be measured simultaneously with the same column because of interference by other absorbing materials. The cortisol and dexamethasone peaks were collected (Rt values 9.6 and 8.1 min, respectively), rechromatographed on another column system, and the concentration was determined. Precision and accuracy of the present method were within the range commonly achieved by other methods established for both steroids. Clinical utility of the present method was evaluated by measuring urinary cortisol and 6 beta-hydroxycortisol in normal subjects, in Cushings syndrome and disease patients, and in patients receiving cortisol therapy.

The molecular abnormality of albumin Niigata: 269 Asp → Gly

A slow albumin variant was isolated from the serum of a patient with bisalbuminemia. Reverse-phase peptide mapping revealed a single altered peak when tryptic digests of the normal and variant albumin were compared. After rechromatography and amino acid analysis, a sequence of Tyr-Ile-Cys-Glu-Asn-Gln-Gly-Ser was obtained for the mutant peptide, while a sequence of Tyr-Ile-Cys-Glu-Asn-Gln-Asp-Ser was obtained for the normal peptide. This establishes the mutation as 269 Asp----Gly and the new albumin has been named albumin Niigata.

Reference limits of bone and liver alkaline phosphatase isoenzymes in the serum of healthy subjects according to age and sex as determined by wheat germ lectin affinity electrophoresis

Wheat germ lectin affinity electrophoresis was employed for quantitating the bone and liver isoenzymes of alkaline phosphatase (EC 3.1.3.1) in serum and for determining the reference limits of each isoenzyme activity in 488 healthy individuals. Bone phosphatase activity was detected even after bone growth, accounting for 60-70% of the total activity. An increase in bone phosphatase activity occurred in older females, but there was a decrease in older males. Liver phosphatase activity gradually increased with age in both sexes, males showing higher activity than females at all ages. Wheat germ lectin affinity electrophoresis of serum alkaline phosphatase is a simple and useful method for quantitating bone and liver alkaline phosphatase activities.

Sugar chain heterogeneity of bone and liver alkaline phosphatase in serum.

Fractionation of bone and liver alkaline phosphatase (EC 3.1.3.1; ALP) in serum by serial lectin affinity chromatography has demonstrated differences in the sugar chain structure of bone and liver ALP in serum from that previously reported in the corresponding tissues, with a lower content of high mannose or hybrid-type sugar chains and a higher content of biantennary complex-type chains. Furthermore, the bone and liver ALPs were found to differ in the latter with the bone fraction showing a greater content of fucose residues.

Age- and sex-related differences in urinary excretion of 6β-hydroxycortisol in humans

Abstract A study of normal subjects showed that the 24-h urinary 6β-hydroxycortisol level is age-dependent, with a maximum at 14–20 yr in both men and women. The ratio of 6β-hydroxycortisol cortisol also showed an age-dependent variation, being higher in children than adults. There was a significant difference between boys and girls aged 5–10 yr, but no difference in other subjects. No sex difference was demonstrable in the ratio of 6β-hydroxycortisol cortisol . Examination of circadian rhythm revealed that 6β-hydroxycortisol and cortisol correlated well in children, but showed poor correlation in adults, with a lowered morning peak of 6β-hydroxycortisol.

Isochronal map of chest wall vibration with cardiokymography

Mapping analysis of cardiokymography is helpful in clarifying the relationship between vibratory pattern and recording sites, but needs a highly stable recording technique. We have therefore studied the stability of recording cardiokymograms, and present here a new method for creating stable maps which enables diagnosis of chest-wall vibration abnormalities. Cardiokymograms were recorded on 16 points of the chest wall located at regular intervals horizontally and vertically. The time interval between a R-wave peak of the electrocardiograms and an E-point of the cardiokymograms were measured for all the 16 records. Isochronal maps in 23 healthy subjects were classified into four types. The map of a patient with myocardial infarction was quite different from any one of the four normal patterns and seemed to show a paradoxical movement of the chest wall.

Portable file management system in FORTRAN

It has been generally considered that FORTRAN is inferior to MUMPS and other computer languages in its facility to manipulate files and that it is not satisfactorily competent to handle databases. However, it is our finding that FORTRAN is the most widely used language in patient data management systems discussed in recently published papers. Therefore, it now seems appropriate to review and evaluate the validity of this general belief. The objective of this study is to establish a file management system in FORTRAN 77 for use in the construction of a clinical database. Comparative study is conducted on several problems associated with file manipulation, that is, time requirement for file access and access methods for data retrieval.

Evaluation of arterial pulse wave velocity on the basis of multivariate analysis

Two sets of photoplethysmographs were used to record arterial pulse waves. A linear predicting function that could predict pulse wave velocity (PWV) by means of age and blood pressure was obtained, and the values computed by this function were subtracted from the measured PWVs. These residuals of the normal subjects were compared with those of the patients in each disease category. The difference between the residuals of normal subjects and those of the patients was not significant.<<ETX>>

Three Families with Fast Bisalbuminaemia

We describe three cases of familial (SH, AY, and TM) fast-type bisalbuminaemia identified from 300 000 electrophoretic strips screened during the past six years. The immunological antigenicity and chemical composition of the isolated fast and normal albumins were essentially indistinguishable in all three cases. Detailed analyses on one of them (TM) by circular dichroism and fluorescent spectra measurements indicated that there was a marked change in the environment of the single tryptophan residue of the fast albumin, suggesting an alteration in the tertiary structure of the molecule. It is likely that this abnormality of the tertiary structure modulated (perturbed) the distribution of electric charges on the protein surface and thus changed its electrophoretic mobility.