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We have developed hybridoma cell lines, each of which secretes a monoclonal antibody (MoAb) to rat renal and hepatic tissue antigens, from Lewis rat with Heymann's nephritis. Three antibodies bind to the brush border of proximal tubular epithelium (BB), one in a fine net-like pattern (no. 3- 11), another one in a coarse granular pattern (no. 1-8) and the third one in a typical pattern (no. 3-9), Three antibodies bind to glomerulus in characteristic patterns but not to BB. After repeated intravenous injections of MoAb (no. 3-11), granular mesangial deposits of rat IgG were observed and of MoAb (no. 1-8), fine granular deposition along capillary walls. These monoclonal autoantibodies should be of value in research on the mechanism of autoimmune membranous nephropathy. [ABSTRACT FROM AUTHOR]

An analytical method of ethephon in feeds by GC-FPD was developed. Ethephon was extracted with ethyl acetate-hydrochloric acid (100 : 1) from feed samples. The extract was treated with added trimethylsilyldiazomethane in acetone-acetic acid (99 : 1) and this methylation procedure was repeated three times. Methylated ethephon was cleaned up on a graphitized carbon mini-column and a silicagel mini-column, and determined by GC-FPD. A method performance study in 7 laboratories was conducted with three kinds of samples spiked with ethephon at 10 mg/kg, 1 mg/kg and 0.5 mg/kg. The recovery of ethephon ranged from 81.8% to 90.8% (the reproducibility standard deviation (RSDr) were within 11%) and HorRat values were 0.58 to 0.94. The limit of detection (S/N≧3) and the limit of quantitation (S/N≧10) of ethephon in samples except hay were 0.02 mg/kg and 0.05 mg/kg, respectively. In the case of hay, the corresponding values were 0.2 mg/kg and 0.5 mg/kg, respectively.

I have been working in the field of laboratory medicine for 45 years including the period of working as a physician. During my career, some of my dreams in laboratory diagnosis have been realized, and some have not. Although the issues changed with time, our basic philosophy has not changed at all. In this session, I would like to discuss "my philosophy of laboratory medicine", that is, what I have intended and what I have done. First, I describe the role of laboratory medicine in the field of clinical medicine. Second, I discuss the value of laboratory tests especially for clinical diagnostics. Third, I summarize the history of clinical pathology with reflection of the past, and my anticipation for the future. Fourth, I mention about the task and responsibility of the laboratory technicians and clinical pathologists, in view of their common purpose, clinical diagnosis. Finally, I discuss the future aspect of laboratory medicine.

We used bone marrow smears to examine how immunohistochemical identification of PCNA (proliferating cell nuclear antigens) which markers of proliferation, works for various hematopoietic cells in hematological disorders. Short-term fixation with buffered acetone formalin allowed us not only to observe morphosis properly but also to identify the occurrence of PCNA in erythroblasts, megakaryocytes, plasma cells and blasts. For blasts of leukemia patients, we could distinguish more positive cases from less positive ones regardless of the increase or decrease of cells. A change in PCNA due to the existence of neoplastic cells was also observed. These findings suggest that in the future, analyzing the occurrence of PCNA in hematopoietic cells can provide us with immunohistochemically important information on diagnosis of malignancy and follow up.

Serum cholesterol levels were determined in 5,843 normal subjects aged zero to seventy years. The accuracy of our assay method was checked by Standard Reference Material 909 and Certified Reference Serum, both of which were supplied by the National Bureau of Standards. Reference values of serum cholesterol were confirmed for each age group. That of male subjects in the 15-20 year age group was within a range of 109-203 mg/dl, and that of female subjects in the 20-25 year age group was within a range of 133-215 mg/dl. Therefore, among normal subjects, the above-mentioned age groups had the lowest serum cholesterol levels. Serum cholesterol levels increased with age in both male and female subjects. The upper limit of cholesterol levels was 248 mg/dl for males in the 50-60 year age group and 284 mg/dl for females in the same age group. We observed the necessity of paying consideration those changes which occur with aging, in the determination of reference values of serum cholesterol. Our findings also showed that serum cholesterol levels remained nearly constant in male subjects of all age groups over a period of 25 years. However, we found mean levels in female subjects in the 50-70 year age groups to be significantly elevated, when compared with those observed in persons in the same age group of 25 years previous. We also found that the mean cholesterol level in girls aged 12 years was higher than that of boys the same age.

The expressions of carbohydrate antigens were examined with panel of specific anti-carbohydrate monoclonal antibodies (MAbs) on human lung cancer tissues. The MAbs used were SH1, SH2, SNH3, AH6, CA3F4, TKH2, TKH6 and TKH5 which define Lex, dimeric Lex, sialosyl Lex, Ley, Lea, sialosyl Tn, Tn and fucosyl GM1, respectively. Paraffin-embedded tissue sections (30 squamous cell carcinomas, 30 adenocarcinomas and 27 small cell carcinomas) were tested by immunohistological staining. Evaluation of stained specimens was performed by taking mean value of scores evaluated by three independent examiners. In squamous cell carcinoma, expression of Ley, sialosyl Tn, sialosyl Lex and Lea was significantly higher than other antigens. Lex was also expressed especially in keratinized tissues. In adenocarcinoma, Lea was expressed most remarkably. Sialosyl Lex, Ley, sialosyl Tn were also highly expressed in malignant cells. There was no significant difference in staining patterns between well and poorly differentiated adenocarcinomas. Sialosyl Tn and sialosyl Lex were positive in morphologically normal mucous glands adjacent to tumors. In small cell carcinomas, Ley was expressed more than other types of tumors, whereas Lex and sialosyl Tn were less than others. Tn antigen was observed throughout adenocarcinomas, squamous and small cell carcinomas in a relatively weak manner. Dimeric Lex and fucosyl GM1 antigens were not detected. Most of normal lung sections showed negative staining with those MAbs. These findings indicate that there are differential expressions of certain carbohydrate antigens in different types of human lung cancers based on their origins.

In spite of the marked advances of laboratory medicine, diagnostic parameters or tools for the pancreatic disorders were now in obscure. However, advances of immunochemical procedures, especially advances of monoclonal antibodies and high performances of immunoassay techniques make it possible to reveal the chemico-pathological status of pancreatic disorders. Moreover, advances of ultrasonic echography and ultrasonically guided biopsy more than make up for the defects of biochemical and immunochemical information. In this symposium, these advances of diagnosis for pancreatic disorders will be studied and discussed.

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.

Analytical and clinical evaluations were made on the measurement of prostatic acid phosphatase (PAP) and prostate specific antigen (PA) by a fully automated enzyme immunoassay system. Results concerning reproducibility, recovery and sensitivity were good. PAP values by this method correlated well with those obtained by radioimmunoassay. PA values by this method were higher than those obtained by other enzyme immunoassays, although the correlation coefficient was high. PAP, PA and gamma-Seminoprotein (gamma-Sm), another prostatic tumor marker, were all poorly correlated to one another. Normal upper value, 2 SD of 720 healthy males was 1.2 ng/ml for PAP, and 3.7 ng/ml for PA. Positive ratios of these tests in 31 patients with prostatic carcinoma were high at advanced stages, and low at early stages. ROC (receiver operating characteristics) curve analysis on patients with prostatic carcinoma and benign prostatic hypertrophy indicated that PAP and PA were more effective than gamma-Sm for the differential diagnosis of prostatic carcinoma, and that the clinical cut off was 2.0 ng/ml for PAP, 7.4 ng/ml for PA and 4.0 ng/ml for gamma-Sm.

Inflammation is a local reactive process against tissue destruction. In inflamed tissue, there are cell reactions such as migration of leukocytes and mobilization of macrophages, and vascular reactions caused by various inflammatory agents. At this symposium, we discussed the recent trend of laboratory tests that might be sensitive indicators of inflammation. Although most of these indicators are nonspecific for diagnosing diseases, some of them might be important in the study for the pathogenesis and for planning treatment. First Dr. Kasahara spoke on the topic of interleukin 1, one of the most important agents that mediated the tissue reaction. The next speaker reported about a classical acute phase reactant, CRP, and a new indicator, SAA. Then there were presentations of two papers on leukocytes and elastase in granulocyte, respectively.

We compared the in vitro antibacterial activities of ceftizoxime (CZX), cefotaxime (CTX), cefmenoxime (CMX), cefoperazone (CPZ), ceftazidime (CAZ), latamoxef (LMOX) and cefotetan (CTT) against 2,729 strains of 11 organisms freshly isolated from 10 medical institutions in Japan between June 1983 and January 1984 and obtained the following results: Against S. pyogenes, LMOX and CTT, which have the methoxy group at the 7 position, were less active than the other drugs. LMOX inhibited 80% of S. pyogenes at 0.78 micrograms/ml; CTT, at 1.56 micrograms/ml; but CZX and CTX inhibited 100% at 0.025 micrograms/ml or lower; CMX, at 0.05 micrograms/ml; and CPZ and CAZ, at 0.20 micrograms/ml. Against H. influenzae, E. coli, K. pneumoniae, P. mirabilis and indole-positive Proteus, these test antibiotics, especially CZX, CTX and CMX, which have the aminothiazolyl methoxyimino group, were potently active. Against S. marcescens CZX and CAZ were more active than the other drugs and against P. aeruginosa CAZ was more active than the other drugs. The test organisms did not tend to acquire resistance to these cephems when our results were compared with the results obtained at the development period.

This study included 35 patients with liver cirrhosis, 23 patients with hyperthyroidism, 12 with hypothyroidism, and 2 with other endocrine disorders. In the various endocrine disorders an appreciable amount of triiodothyronine (T3) was excreted into the urine but the daily excretion was fairly constant in each patient. Urinary excretion of T3 was negligible or depressed in hypothyroidism, but increased with a rise in the serum level of T3. Serum and urinary T3 decreased in liver cirrhosis, but the serum thyroxine (T4) level was within the normal range. When the cirrhosis patients were divided into 3 groups according to the urinary excretion of T3, a decrease of urinary T3 was associated with a decrease in the serum levels of T3 and free T3. An increase of serum thyroid-stimulating hormone (TSH) either before or after injection of thyrotropin-releasing hormone (TRH) was inversely correlated with a decrease of serum and urinary T3. The decrease of serum and urinary T3 was correlated with the magnitude of lh a decrease in the serum levels of T3 and free T3. An increase of serum thyroid-stimulating hormone (TSH) either before or after injection of thyrotropin-releasing hormone (TRH) was inversely correlated with a decrease of serum and urinary T3. The decrease of serum and urinary T3 was correlated with the magnitude of lh a decrease in the serum levels of T3 and free T3. An increase of serum thyroid-stimulating hormone (TSH) either before or after injection of thyrotropin-releasing hormone (TRH) was inversely correlated with a decrease of serum and urinary T3. The decrease of serum and urinary T3 was correlated with the magnitude of liver damage as judged by indocyanine green retention and a decreased urinary excretion of cyclic adenosine 3',5'-monophosphate. In vitro experiments indicated that rat liver, as compared to the kidney, heart and skeletal muscle, strongly converts T4 to T3, but this activity is greatly reduced by liver damage induced by ligation of the bile duct. It is suggested that patients with liver cirrhosis are, to some extent, in a state resembling subclinical hypothyroidism because of inability of the liver to metabolize a sufficient amount of T3 from T4.

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.

A study of normal subjects showed that the 24-h urinary 6 beta-hydroxycortisol level is age-dependent, with a maximum at 14-20 yr in both men and women. The ratio of 6 beta-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 6 beta-hydroxycortisol/cortisol. Examination of circadian rhythm revealed that 6 beta-hydroxycortisol and cortisol correlated well in children, but showed poor correlation in adults, with a lowered morning peak of 6 beta-hydroxycortisol.

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.

A comparative study between normal groups showed that the 24-h urinary cortisol level is age-dependent both in men and women. The comparison between men and women with age-matched groups revealed a significant difference in the lower age groups up to 20-24 years. When urinary cortisol values were corrected for creatinine excretion, the age-related variation was eliminated to some extent, but not completely. The correction could correct the sex-related difference in the lower age groups, but it brought about another sex-related difference in the advanced age groups. There was no remarkable age- or sex-related difference in the circadian rhythm.

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.