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The metabolism and nutrition section of JSLM2015 was constructed following the previous version JSLM2012. The examinations of dyslipidemia and glucose metabolism and weight loss and obesity were described in the first chapter Laboratory data approach and the second chapter Symptoms, respectively. Dyslipidemia, diabetes, gout and hyperuricemia, and osteoporosis were described in the third chapter Diseas- es. This paper provides an outline with a focus on dyslipidemia examination and dyslipidemia in the first and third chapters, respectively. [Review].

Many clinical studies, including Framingham Heart study, have demonstrated that increased LDL choles- terol and decreased HDL cholesterol are associated with the risk of coronary heart disease (CHD), and lipo- protein evaluation is of importance in the prevention and treatment of CHD risk. Lipoprotein analysis methods by high performance liquid chromatography (HPLC) are divided broadly into 2 categories (HPLC with gel-filtration column and with anion-exchange column), and both methods can determine lipid levels of fractionated serum lipoproteins in a small volume of around 10 pL within 30 min. In contrast, with the gel- filtration HPLC method by Gaussian approximation following the particle sizes, the anion-exchange HPLC method elutes lipoproteins following the ion intensity of the lipoprotein particle surface and the hydrophobic property, and determines cholesterol levels of separated lipoproteins without overlapping lipoprotein frac- tions. We have established a method with anion-exchange chromatography using a 1.0-mL injection volume that 5.2 min takes to assay one sample. The within-day and between-day assay coefficients of variation of lipoprotein cholesterol values were 0.33-4.31 and 2.37-9.19%, respectively. Previous studies generating fractionated lipoprotein cholesterol data using anion-exchange HPLC revealed the clinical significance in a variety of settings and, in Japan, this rapid anion-exchange HPLC assay was approved as a medical examina- tion covered by health insurance for medical service fees in 2013, and the anion-exchange HPLC method for lipoprotein fraction cholesterol measurement has the potential to offer further clinical benefit. [Review].

When diagnosed with hyperlipidemia, type classification should be performed first. Next, in conjunction with a medical interview about any family history of hyperlipidemia, secondary hyperlipidemia should be dis- tinguished and the treatment of underlying disease should take priority. Administering LDL-lowering ther- apy with HMG CoA reductase inhibitors (statins) to patients with a hypothyroid state without a differential diagnosis of secondary hyperlipidemia may be associated with a high risk of myopathy and liver dysfunction. This symposium was organized to shed light on the etiology of secondary hyperlipidemia and abnormalities of lipid laboratory tests because medical care for secondary hyperlipidemia is of marked significance. Lipid metabolism disorders induced by diabetes, endocrine diseases, liver diseases, renal diseases, and drug thera- pies were presented by respective specialists. All of the contents will benefit medical care and the examina- tion of dyslipidemia in the future, and they are also useful for routine medical activities. [Review].

The Japanese Society of Laboratory Medicine has been running its own Medical Safety Committee, and holding a symposium on medical safety during the annual meeting every year. Adopting a team approach to medicine plays a critical role in the development of medical safety culture and advancing medical safety in clinical practice. The infection control team plays a major role in team medical care. This time, the review of safety management from infection control perspective was discussed in the medical safety symposium, which is hoped will help advance medical and patient safety, leading to improvements in the quality of medical care.

We established a method for measurement of the cholesterol concentrations of high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), intermediate density lipoprotein cholesterol (IDL-C) and very low density lipoprotein cholesterol (VLDL-C) by using high performance liquid chromatography (HPLC) with anion-exchange column. The HPLC method has been covered by insurance in 2013, and HLC-729LPII (LPII) system constructed by this method has come on the market in 2014. We evaluated the fundamental precision data of lipoprotein cholesterol values measured by HLC-729 LPII. The within-day and between-day assay coefficients of variation of lipoprotein cholesterol values were 1.4-10.7 (%CV). The lipoprotein profiles of patients with type 2 diabetes mellitus (T2DM, n = 60) without dialysis therapy were measured by LPII. HDL-C obtained by LPII was highly correlated with that obtained by direct assay. LDL-C obtained by LPII was highly correlated with those obtained by direct assay and calculated by Friedewald's formula. In addition, IDL-C obtained by LPII was negatively correlated with estimated Glomerular Filtration Ratio (eGFR). These results suggest that the new HPLC method can be applied to estimate lipoprotein profile of T2DM patients. Particularly, IDL cholesterol may be useful for the evaluation of impaired lipid metabolism in T2DM patients without dialysis therapy, but it remains to be cleared.

The Japanese Society of Laboratory Medicine has been running its own Medical Safety Committee, and holding a symposium on medical safety during the annual meeting every year. The activities based on a team approach to medicine play a critical role in the development of medical safety culture and the driving forward of medical safety in clinical practice. The best medical practice involves cooperation among information-sharing medical staff from a variety of professions, providing medical care based on medical safety, which patients hope for. It is hoped that the present symposium on medical safety can help drive medical safety into the future.

The LDL-C level measures with homogeneous (direct) assays in almost of clinical laboratories. Several reports however showed differences in measured values among the assay reagents. We investigated the differences in LDL-C values among direct assays and Friedewald formula (F-f) in 58 LP-X positive serum samples from jaundice patients by comparing LDL-C values measured by anion-exchange chromatography (AEX-HPLC), largely comparable to ultracentrifugation method. Changes in LDL-C values during the treatment of 8 patients were also investigated. Direct assay reagents from Sekisui Medical (S-r), Denka-Seiken (D-r), Wako Chemical (W-r), and Kyowa Medics (K-r) were used for comparison. F-f, S-r, and D-r correlated with AEX-HPLC with r values < 0.6 while W-r and K-r correlated with AEX-HPLC with r-vales > 0.6. Two samples in which F-f values provided 500 mg/dL plus bias to AEX-HPLC (LDL-C value of 220 mg/dL) demonstrated increased levels of IDL-C before treatment. LDL-C values (S-r and D-r) of the 2 samples were relatively high and near to F-f data while LDL-C values (W-r and K-r) were relatively low and close to AEX-HPLC data. The jaundice treatment decreased LDL-C values (S-r and D-r) and converged to 220 mg/dL, indicating that S-r and D-r might react markedly to IDL. These changes were consistent with decreases in serum free cholesterol and phospholipid in support of LP-X. By contrast, W-r and K-r data showed upward tendency and also converged to 220 mg/dL. These results suggest that LDL-C direct assay reagents would be classified into 2 groups with respect to the reagent reactivity to LP-X.

Chronic kidney diseases (CKD) have been cited as major risk factors not only for endstage renal failure, but also for the development of cardiovascular diseases and death. In the former criteria for CKD diagnosis (NKF K/DOQI, 2002), estimation of the severity of CKD was simply based on GFR, in order for it to be widely and easily understood by general physicians and patients. Therefore, the use of the CKD guideline without information on the causes and grades of albuminuria was limited for estimation of the prognosis. The revised guideline for CKD diagnosis (KDIGO CKD guideline 2012), with the disease category specified for diabetes mellitus and the different scoring system of microalbuminuria, is presently being effectively utilized by nephrologists as well as general physicians. In this symposium, to advice understanding of the causes and evaluation methods of CKD, speakers were invited to discuss topics in kidney pathology, urine sample examination, urinary biomarkers, and GFR.

We conducted a questionnaire survey regarding the current activities for protecting patients' privacy and the security of information systems (IS) related to the clinical laboratory departments of university hospitals, certified training facilities for clinical laboratories, and general hospitals in Yamaguchi Prefecture. The response rate was 47% from 215 medical institutions, including three commercial clinical laboratory centers. The results showed that there were some differences in management activities among facilities with respect to continuing education, the documentation or regulation of operational management for paper records, electronic information, remaining samples, genetic testing, and laboratory information for secondary use. They were suggested to be caused by differences in functions between university and general hospitals, differences in the scale of hospitals, or whether or not hospitals have received accreditation or ISO 15189. Regarding the IS, although the majority of facilities had sufficiently employed the access control to IS, there was some room for improvement in the management of special cases such as VIPs and patients with HIV infection. Furthermore, there were issues regarding the login method for computers shared by multiple staff, the showing of the names of personnel in charge of reports, and the risks associated with direct connections to systems and the Internet and the use of portable media such as USB memory sticks. These results indicated that further efforts are necessary for each facility to continue self-assessment and make improvements.

The Japanese Society of Laboratory Medicine has been running its own Medical Safety Committee, and holding a symposium on medical safety during the annual meeting. The medical world is filled with a considerable amount of personal information, including genetic information, the ultimate personal information. We, as medical staff, have to manage such personal information not only in times of peace but also during disasters or emergency situations. In Japan, the Act on the Protection of Personal Information is currently being implemented, but a number of problems remain. Human beings have entered the information technology era, including electrical medical record systems, which is useful for research and education besides medical practice. This is why personal information must be more effectively protected from leakage, misconception, and abuse. We should create a sound system to manage personal information, with the spirit of protecting patient information that originated from the Oath of Hippocrates.

Measures against atherosclerotic diseases, accounting for one-third of the causes of death in the Japanese population, are critical for increasing human healthy life expectancy. In Japan, guidelines for the prevention of atherosclerotic cardiovascular diseases in 2012 and a dyslipidemia therapy guide were published by the Japan Atherosclerosis Society, and an outline of the clinical laboratory field, including serum lipid measurements, was also described in guidelines of the Japanese Society of Laboratory Medicine in 2012. Calculation using the Friedewald formula is recommended for LDL-cholesterol measurement, but the calculated LDL cholesterol values are not applicable in samples that are postprandial or with serum triglyceride (TG) levels of more than 400 mg/dL, and, if this is the case, non-HDL cholesterol (total cholesterol minus HDL cholesterol) should be evaluated. Such descriptions of LDL-cholesterol in these guidelines may be attributed to problems of homogenous LDL cholesterol assay reagents, including insufficient standardization and dispersion of LDL cholesterol values using a variety of reagents. Their accuracies have been questioned, especially in diseased subjects from the report of a joint U.S.-Japan study (Clin Chem 2010; 56: 977-86), and, subsequently, Miida T and Yoshida H et al. reported that the LDL-cholesterol homogenous assay agrees with the LDL cholesterol reference method (BQ) in non-diseased subjects, but exhibits positive bias for subjects with hypertriglyceridemia in diseased subjects with some reagents (Atherosclerosis 2012; 225: 208-15). Future improvements and advanced standardization of the LDL cholesterol homogenous assay are expected to greatly improve the credibility of serum lipid testing for the prevention and clinical treatment of atherosclerotic cardiovascular diseases.

The revised version of the guideline JSLM 2012 was published in December 2012 by the Japanese Society of Laboratory Medicine. This version included new sections, such as blood gas analysis, sleep apnea syndrome, interstitial lung disease, liver and pancreas cancer, chronic kidney disease, acute kidney disease, gout and hyperuricemia, bone metabolism abnormality, malignant lymphoma, and rheumatoid arthritis. The guideline committee was composed of specialists in each field of internal medicine, who were responsible for selecting and requesting the authors and also in promoting and proofreading the manuscripts. In special program I at the 60th annual meeting, each specialist gave lectures concerning the points of the revision in their fields. The questionnaire surveys were performed using FAX or the Internet. Analysis of eighty-seven (2.5%) responses from 3,500 individuals/facilities, which were sent the guideline, revealed that this guideline was graded as excellent by 38 readers, fair by 42, and average by in 6. Significant opinions on the guideline were obtained from the readers, and they will be the bases for the next revision. The main subjects of this guideline were confirmed to be the residents and general physicians, by whom it is hoped the routine laboratory tests will be properly utilized. Therefore, based on the section of 'approach of the laboratory test results', which is a representative characteristic of this guideline, the sections of 'symptoms' will be fulfilled for the next version. This guideline needs to be periodically revised with advances in medicine.

Since the launch of the "National Joint Action of Risk Management" back in May, 2008, as a participating organization, The Japanese Society of Laboratory Medicine has been running a Medical Safety Committee of our own. As part of our awareness education activities, our society has been holding symposia on medical safety at our annual meetings. In clinical practice, misidentification of patients is directly connected to serious medical mistakes. Although clinical practice itself must be safe for all, the actual clinical setting poses many hazards that could lead to medical professionals making errors. Since misidentification of patients regarding obtained specimens such as blood or physical function exams and misinterpretation of the exams itself can result in a serious outcome, every institute is using ingenious ways to keep close checks to prevent such misidentifications from happening. In this symposium, lectures were presented in the following subjects: A review on countermeasures against specimen misidentification in pathological exams in hospitals using electronic patient verification systems to prevent misidentification of patient blood samples, and hospitals where such systems are not employed; an introduction of countermeasures against specimen misidentification in pathological exams; and lastly, characteristics and limitations of human abilities, and process management, entitled "Human Factor Approaches to Medical Safety".

The pseudohyperkalemia in thrombocytosis is assumed to be due to potassium released from blood cells during blood clotting as reported previously, but its mechanisms remain to be cleared. Although plasma potassium measurements with blood collection tubes containing heparin are performed in many hospitals to avoid pseudohyperkalemia, the burden on patients may come out with further blood sampling by another heparinized tube. Taken together, we investigated laboratory data possibly involved in pseudohyperkalemia in 184 samples from patients with myeloproliferative neoplasma (MPN), and studied estimation capability for true values of serum potassium, driving a correction formula by means of several laboratory data to explain the difference of measured potassium values (K-difference: serum value minus plasma value). Platelet count and mean corpuscular volume (MCV) were adopted as significant variables correlated to K-difference as a result of multiple regression analysis. A correction formula was driven by multiple regression equation with these two variables as follows: y = 0.0006 x 1+0.0004 x 2-0.177 (r= 0.885; x 1, platelet count; x 2, MPV). The correction formula was considered to be useful for estimating the true value of serum potassium in samples from patients with MPN because the corrected serum potassium value correlated highly with plasma potassium value (r = 0.885). These results propose that true values of serum potassium can be estimated by the correction of measured serum potassium values with platelet count and MCV, suggesting that not only quantitative factors but also qualitative factors may be involved in pseudohyperkalemia.

Lowering low-density lipoprotein (LDL) cholesterol (LDL-C) can reduce the risk of cardiovascular disease (CVD) by approximately 30%, and the remaining 70% should be the second front of CVD risk reduction. Such residual risks include high triglyceride (TG) concentrations and low levels of high-density lipoprotein (HDL) cholesterol (HDL-C) in terms of dyslipidemia. TG-rich lipoproteins are heterogenous and composed of a variety of subfractions, all of which are not necessarily relevant to atherosclerosis and CVD risk. However, remnant lipoproteins, TG-rich lipoproteins, are atherogenic and related to CVD risk. Two different methods (RLP-C and RemL-C) have been developed to measure cholesterol levels of remnant lipoproteins. Although there is a difference in affinity to intermediate-density lipoprotein (IDL) between the two methods, they may be better qualified as biomarkers of CVD risk than TG itself. TG measurements play a certain role in the evaluation of CVD risk, but the remnant lipoprotein cholesterol measurement can provide better screening for patients at high CVD risk than TG and may be a useful examination in both quantity and quality.

I started my life as a medical doctor at Amagasaki Prefectural Hospital after graduation from the Faculty of Medicine, Kyoto University in September 1971. I joined the newly established section of nephrology in the second year. The chief was Dr. Kazuro Kanatsu who had just moved from Kyoto University at the time of the campus disturbances. Dr. Kanatsu not only oriented me in clinical nephrology, but also guided me in medical research. I used to go to the laboratory of Dr. Tadao Tamura, Kyoto University once a week to learn renal biopsy study. In 1977, I entered the Department of Pathology, Postgraduate School of my university to learn immunopathology from Prof. Yoshihiro Hamashima. In the second year, I was willingly involved in the research group on murine SLE organized by the newly invited associate professor, Dr. Toshikazu Shirai, who taught young researchers such as myself how to consider, practice and enjoy experiments. In 1982, I went abroad to Prof. Peter Miescher, University of Geneva, who was a friend of Prof. Hamashima and organized immunopathology research groups. In the laboratory of Prof. Shozo Izui, I performed an isoelectric focused study on anti-DNA antibodies in lupus-prone mice and identified the pathogenetic role of the clonal expansion of autoantibodies. After 3 years, I came back to the 3rd Division of Internal Medicine of my university. Meanwhile, Prof. Chuichi Kawai guided me to go back to Prof. Hamashima's Pathology Department, where I helped young doctors publish a series of papers, including studies on SLE and a murine model of IgA glomerulonephritis. Later, I was obliged to leave the Pathology Department, and moved to Himeji National Hospital in 1992 as a clinical nephrologist by the invitation of the Director, Dr. Tamura. At that time I was very much encouraged by Prof. Shirai at Juntendo University, who gave me a letter with an old saying "Jinkan itarutokoro seizan ari". After 3 years, I moved to Kitano Hospital, Osaka, where I learned up-to-date information and techniques in clinical nephrology. From this hospital, I published a paper in Kidney International entitled, "Mesangiolytic glomerulopathy in severe congestive heart failure", based on the autopsy cases collected at the Pathology Department. This paper became a milestone in starting to study the role of chronic hypoxia in CKD. In 1999, I was elected as a professor of the Department of Clinical Laboratories, Faculty of Medicine, University of Fukui. In Fukui, I could extend my hypoxia study to cellular levels and diabetic mouse experiments in collaboration with Dr. Kimura, Dr. Li, Dr. Takahashi and many other doctors and technicians. When overviewing my research history, I realize that I was fortunate to be involved at the starting point of every laboratory with energetic mood and that I was supported and helped by many people.

Diurnal variations in serum iron concentration were examined to investigate the influence of sampling time in hemodialysis (HD) patients and healthy subjects. The serum iron concentration and TIBC of HD patients decreased significantly (p<0.01, p<0.01 respectively) compared to those of healthy subjects. Inversely, the serum ferritin concentration of HD patients increased significantly (p<0.01) compared to that of healthy subjects. These findings show the disturbance of iron transport system: under such condition intracellural iron transition out into peripheral blood stream is low in HD patients. Serum iron concentration in samples collected in the evening decreased significantly both in HD patients (p<0.05) and in healthy subjects (p<0.01). Diurnal variations in serum iron concentration reveal almost similar decrement in both groups. In HD patients, serum iron concentration of the blood samples collected on the third day morning after HD and second day morning after HD was examined to see the influence from changes of circulating plasma volume. The serum iron concentration and Hct value in the second day sampling increased significantly compared to the third day sampling (p<0.01, p<0.01 respectively). In addition, the serum iron concentration corrected by Hct in the second day sampling increased significantly (p<0.01) compared to the third day sampling. We conclude from our results that diurnal variations of serum iron concentration vary in sampling time in HD patients as well as in healthy subjects. We also deduct that there may be other factors concerning changes in circulating plasma volume.

Frequency domain analysis of heart rate variability (HRV) is used for the evaluation of autonomic activity. Non-linear domain parameters from HRV are also considered useful. However, properties of the latter have not yet been clearly characterized. Therefore, we studied the relationships among the frequency domain and non-linear parameters from HRV. Continuous Holter electrocardiographic monitoring was conducted on 43 healthy female medical staff including laboratory technologists and nurses during an 8-hour working period in our hospital. Low and high frequency components (LF and HF, respectively) of the frequency domain, recurrence rate (REC%) on recurrence plot analysis, scaling exponents al and a2 on detrended fluctuation analysis, and approximate entropy (ApEn) were obtained from HRV. Both the LF/HF ratio and HF were correlated with al and ApEn. REC% was correlated with ApEn and alpha2, whereas alpha2 was correlated only with REC%. Although autonomic parameters from the frequency domain are closely related with some of the non-linear parameters, it is suggested that a2 and REC% reflect different physiological activities.

Safety management is essential for providing patients with medical services. Our hospital, opened at Kashiwa in 1987, has been building up systems and taking a number of steps to reduce the blood collection related problems, including venipuncture-related infection, nerve injury, and vasovagal reflex with syncope in accordance with guidelines for the standard method of venipuncture blood collection. We also have made efforts for improving medical services, including reductions in patient waiting time and prevention of patient misidentification, medical test malpractice, and patient privacy. However, ultimately, it is of obvious significance to educate and train communication skills for humanity and friendly kindness because most of medical accidents are basically attributed to communication errors between patients and medical staff.

Thioredoxin (TRX) is a redox-regulating protein, induced in response to oxidative stress. The function of TRX in the urine is unknown. We show here that urinary TRX begin to increase within one hour and peaks within two hours after ischemia reperfusion of mice. Serum levels of TRX are not changed by the ischemia/reperfusion. In a time-dependent study of immunohistochemistry, TRX appears diffusely in the tubular cytosol in sham-operated mice. On the other hand, immediately after renal ischemia/reperfusion, TRX become to eccentrically-locate in the apical side of the tubular cytosol, and then TRX is detected only in the urinary lumen. In contrast, when we examine the immunolocalization of glutaredoxin, which is a member of the TRX superfamily, we find that the immunoreactivity is unchanged after renal ischemia/reperfusion. Northern blotting and in situ hybridization show that epithelial cells constitutively express TRX mRNA but neither expression levels nor distribution are altered by ischemia-reperfusion. An overexpression of hTRX in transgenic mice attenuates the reperfusion injury. These data suggests that TRX is produced in tubular cells in a steady state. The increase in the urine after ischemia-reperfusion is not mediated by a de novo induction of TRX mRNA but by a discharge of TRX protein from tubular epithelial cells. TRX is useful for the diagnosis of AKI in association with oxidative stress.

Lipoprotein analysis methods employing high-performance liquid chromatography (HPLC) are divided broadly into 2 categories (HPLC with a gel-filtration column, and with an anion-exchange column), and both methods can determine lipid levels of fractionated serum lipoproteins of small amounts around 10 microl within 30 min. The former HPLC method separates lipoproteins based on the particle sizes, and larger-sized lipoproteins are eluted earlier, but this gel-filtration HPLC method determines lipid levels of lipoprotein fractions by Gaussian approximation. The latter HPLC method elutes lipoproteins based on the ion intensity of the lipoprotein particle surface and the hydrophobic properties, and determines cholesterol levels of separated lipoproteins without overlapping lipoprotein fractions. A large amount of research data on lipoprotein cholesterol levels measured using the anion-exchange HPLC method have been reported in patients with various diseases (diabetes, dyslipidemia, coronary heart disease, or chronic renal failure), and the anion-exchange HPLC method deserves a position of much greater clinical significance.

Ever since oxidized low density lipoprotein (LDL) was first identified in human atherosclerotic lesions, probucol was found to suppress the development of atherosclerosis in animals, and LDL incubated with vascular cells underwent oxidative modification, the so-called oxidative modification hypothesis of atherogenesis was nurtured, and a number of significant pieces of evidence have accumulated to suggest the underlying contributions of oxidized LDL to the development of atherosclerotic diseases. Recently, oxidized LDL in blood circulation can be measured, and plasma-oxidized LDL is a well-known risk marker of cardiovascular diseases. Oxidized LDL-mediated macrophage foam cell formation is probably attributed to the lack of down-regulation of oxidized LDL receptors, but high-density lipoprotein (HDL) or apolipoprotein A1-mediated cholesterol efflux from cells is also promoted, probably by oxidized LDL. This may be why macrophage cholesterol accumulation induced by oxidized LDL does not advance unilaterally. However, such a suppressive effect of HDL may be inhibited once HDL undergoes inflammatory or oxidative modification because oxidized HDL could be dysfunctional. Remnant lipoproteins can be taken up by macrophages without oxidative modification in contrast to LDL, and remnant lipoproteins exhibit many atherogenic actions similar to oxidized LDL. However, oxidized remnant lipoproteins do not function as an effective material of foam cells. Recent papers have demonstrated that oxidized phospholipid (PL)/apolipoprotein B, playing a potential role in predicting cardiovascular diseases, correlates strongly with lipoprotein (a) [(Lp (a)] because Lp (a) is susceptible to binding to oxidized PL, and oxidized Lp (a) is also detected in human coronary atherosclerotic lesions. Although many lines of evidence support the oxidized LDL hypothesis of atherosclerosis, we face important problems to be overcome, such as the fact that oxidized lipoproteins may not be relevant to human atherosclerosis because antioxidation treatments with vitamins do not necessarily prevent human cardiovascular diseases. Oxidized LDL could play a role in the initiation of foam cell generation, but become less important in the later stages of atherosclerotic disease. Taken together, we have to keep an open mind regarding the incompletely understood pathogenesis in association with oxidized lipoproteins.

Excess adiposity has been shown to play a crucial role in the development of the metabolic syndrome. Characteristics for dyslipidemia in the metabolic syndrome are elevated fasting and postprandial triglyceride (TG) and decreased high-density lipoprotein-cholesterol (HDL-C). Diacylglycerol(DAG) has been suggested to suppress postprandial hyperlipidemia and promote negative caloric balance by increasing energy expenditure, due to intestinal physiochemical dynamics that differ from triacylglycerol (TAG). Our study (Study 1) demonstrated that DAG suppressed postprandial increase in TG-rich lipoprotein, very low density lipoprotein (VLDL), and insulin as compared with TAG in young male individuals. Further, our another study using the apolipoprotein C-II deficient subject demonstrated that DAG suppressed postprandial increase in VLDL-cholesterol and remnant-like particle-cholesterol compared with TAG, suggesting that DAG suppress postprandial TG-rich lipoprotein independent of lipoprotein lipase. Study 1 also showed that DAG significantly increased plasma serotonin, which is mostly present in intestine and mediates thermogenesis, proposing a possible mechanism for a postprandial increase in energy expenditure by DAG. Our studies presented DAG-mediated amelioration in postprandial TG-rich lipoprotein, insulin, and energy metabolism, indicating the therapeutic application of DAG for the metabolic syndrome.

Dr. Hideyo Noguchi (Noguchi) is the most well-known scientist in Japanese history because of his eventful life and research on syphilis and yellow fever; however, details of his scientific research, especially in the seroimmunological field, performed in the USA have not been recognized. More than 200 papers were published, mostly in English, and about half of them were published in J. Exp. Medicine. Arbitrary evaluation was performed of his research, recognizing the value of his seroimmunological studies. In this lecture, the background at that time and a small part of the contents of 10 papers on serology and biochemistry are introduced and explained, in connection with recent seroimmunology: (1) On snake venom: In the first experiment, under the guidance of Drs. Flexner and Mitchel, many new findings were obtained regarding bacteriolysis, hemolysis, leucolysis, and the toxicity of venoms in relation to antibody and complement. Especially, the bacteriolytic activity of serum induced by venom was disclosed later to be caused by the activation of alternative pathway of complement by a factor termed cobra venom factor. (2) Heat-stable anticomplementary factor: Preheating sera produced an anticomplementary factor, termed protectin. (3) The pleurality of cytolysins (natural antibody) in sera not only of mammals but also of a reptile and an amphibian was identified. (4) Toxin-antitoxin reaction: One of the fundamental analyses of the antigen-antibody reaction. (5) Gel diffusion of antigen and antibody. (6) Various factors affecting the Wassermann reaction, in which the butyric acid method to detect globulin was introduced. (7) A new and simple method for the serodiagnosis of syphilis. (8) A study of syphilis antigen, in which the antigenicity of acetone-soluble lipid was stressed. (9) Pure cultivation of Treponema pallidum. (10) Demonstration of T. pallidum in the brain. More than 40 papers on syphilis were published, for which he was nominated for the Nobel prize 3 times.

Objective: Methicilllin-resitant Staphylococcus aureus (MRSA) is still the most important bacterium for hospital infection control, and is known to exhibit beta-lactam resistance. Moreover, the increase in PBP2'-producing methicillin-resistant coagulase-negaive Staphylococcus (MR-CNS), especially methicillin-resistant S. epidermidis (MRSE) has been problematic. In this study, we investigated the induction of PBP2' by MPIPC, other antibiotics and disinfectants in MRSE., Materials and Methods: The bacterial strains used were MRSE isolated in our clinical laboratory. MRSA-LA 'Seiken' was used for the detection of PBP2'. To investigate induction of PBP2' by MPIPC in MRSE, MRSE was cultured on the medium containing MPIPC at 11 different concentrations from 0.0001 to 6 microg/ml, and PBP2' induction was investigated. Strains in which no induction was noted at a low MPIPC concentration were cultured with other antibiotic discs and discs impregnated with various disinfectants, and PBP2' was detected in colonies that grew around the disc and PBP2' induction was investigated., Results: In the culture on MPIPC-supplemented medium, PBP2' was detected in all strains at 0.01-6 microg/ml. At 0.001 and 0.0001 microg/ml, 8/10 and 4/10 were positive, respectively. Addition of another beta-lactam, particularly cephem antibiotics, induced PBP2' in some strains that were negative at 0.0001 microg/ml. In cultures with disinfectants, inhibition zones were noted, but no PBP2' was induced., Conclusion: PBP2' was induced by a low beta-lactam and was not by disinfectants in MRSE.

Mental arithmetic (MA) tasks have been reported to induce an increase in blood pressure and heart rate by enhancing sympathetic activity. However, there are individual differences in the change of autonomic nervous activity during MA task. In the present study, we quantitatively analyzed EEG changes and autonomic nervous activity using heart rate variability (HRV) during MA task in 38 healthy young volunteers. They were classified into two groups, stress responders and non-responders, according to whether both LF/HF and blood pressure increased above the baseline or not in response to MA. Then autonomic and EEG changes were compared between the two groups. The MA task induced an increase in slow beta power in the stress responders, whereas it induced a decrease in slow beta power in the stress non responders. Further evaluation showed a significant positive correlation between the percent change from the baseline in slow alpha power and that in LF/HF. These results suggest that there are correlations between the autonomic response in an individual and EEG power changes during MA tasks.

Preanalytical issues in laboratory medicine have been recognized to be important but not so much effort have been focused to them. Urinalysis is one of the most susceptible examinations to preanalytical issues because urine samples are collected by patients themselves and so is stool examination too. Midstream urine catch is well-known appropriate procedure for many urinalysis. Surface scratching is an appropriate method of stool occult blood testing for colorectal cancer and also strategies to avoid the contact of stool with toilet water is another important technique. We also report a case of abnormal results of creatinine clearance (Ccr) caused by the urine collection apparatus as an example of preanalytical issue.

Increase in triglyceride (TG) -rich lipoproteins is one of the symptoms clustered in metabolic syndrome and is associated with increased plasma free fatty acid level derived from central obesity and insulin resistance. Increase in triglyceride (TG) -rich lipoproteins is also related to several coronary risk factors such as remnant hyperlipidemia, decreased HDL-cholesterol, elevated small dense LDL, postprandial hyperlipidemia, and hypercoagulability. The first line of treatment for hypertriglyceridemia is the modification of individual life-style, among which, restriction of over-eating and practice of regular exercise are both essential. The consideration of dietary composition, not only the quantity but also the quality of nutrients, such as fat and carbohydrate, and behavior toward diet are also important to manage abnormal lipid profile. Statins, fibrates, nicotinic acid derivatives, and EPA are the drugs recommended for the treatment of dyslipidemias in metabolic syndrome.

Differences in bleeding time and self-monitoring blood glucose (SMBG) level at various skin sites have been scarcely examined in humans. The within-run variation (n=6) of bleeding time in earlobes (Duke's method) ranged from 1 min to 3 min and 30 sec (mean = 1 min and 40 sec), and in day-to-day variation (n= 8), it ranged from 1 min to 3 min (mean = 1 min and 36 sec). Site difference in bleeding time was speculated. In SMBG, firstly, sequential measurement of blood collected from 10 sites in the left forearm was performed, and secondly, comparative measurements of venous blood and blood from finger tips (1-5th), palm (3 sites), forearm (4 sites) and earlobe (1 site) were sequentially performed within 30 min before and after glucose (Trelan-G, 75g) intake. Glucose levels in blood from the fingertips, palm and forearm were generally higher than that of venous blood, and site differences were observed among fingertips, palm and forearm. It was speculated that bleeding time and capillary blood glucose or SMBG level differ among skin sites.

Clinical laboratory staff should provide laboratory data showing or reflecting the exact patho-physiological conditions of patients or examinees (exact data). Intralaboratory quality management and assurance have been improved to satisfactory levels; however, preanalytical processes have been poorly checked and the situation is still unsatisfactory. The improvement of preanalytical issues is essential to provide exact data. Among various issues affecting the data, inadequate sampling has been and will be the major problem to be tackled by laboratory staff. In blood collection, more than 90% of samples are collected by nurses and most urine, feces and sputum are collected by patients themselves. To improve the current situation, regular meetings with other hospital staff and providing information on clinical laboratory medicine are possible ways to make them aware of current problems. Two examples are presented: the clinical laboratory practice of medical students in Fukushima Medical University and the Clinical Laboratory Forum held in 1991-1999, which was performed in collaboration with Fukushima Associations of Nurses, Medical doctors and Laboratory technician and laboratory staff of Fukushima Medical University. Additionally, the effect of freezing and thawing at different temperatures were compared for 30 biochemical items. About half of individual sera showed reduced levels of ALT and LD when frozen at -20 degrees C and thawed at 25 degrees C compared with those frozen at - 80 degrees C and thawed at 37 degrees C.

As the final lecture of Clinical Laboratory Medicine in Fukushima Medical University, I showed some of our findings obtained by the analyses of unexpected laboratory data. Most of these unexpected or unusual data were first noticed by laboratory staffs in charge. Items mentioned were; 1) false negative reaction of immunological fecal occult blood test, induced by the contact of hemoglobin with toilet sanitizers or detergents, 2) highly elevated serum K level, induced by the cold storage of whole blood, 3) false positive reactions in ELISA, induced by the aggregated IgG developed by the heat-pretreatment of sera, 4) panic values developed by the regurgitation of EDTA blood, 5) markedly low values due to the non-mixing of frozen thawed serum sample, 6) precision of the bleeding time in earlobes, and 7) differences in capillary blood glucose levels among forearm, palm and fingers. In the analyses of 6) and 7), the methodology of quality assurance could be introduced, although we realized the difficulty in examining in our physical body. Every issue examined is practically important, and laboratory staffs including physicians should pay more attention on the unexpected laboratory data and know why and how the data are produced.

Objectives: To disclose the current situation of rheumatoid factor (RF) measurement in Japan., Method: A small-scale survey was performed among members of the committee for the improvement of RF measurement in 2003 using manufacturers' RF reference materials valued based on WHO RF reference and pooled sera. A nationwide questionnaire survey was also performed. Questionnaires were sent to 356 educational institutes certified by the Japan College of Rheumatology in December 2004., Results: The small-scale survey showed inter-laboratory differences of measured RF values, especially using RF references, and less in pooled sera. In the questionnaire survey (187 responses, recovery, 53%), qualitative or semi-quantitative RF measurement methods were used in a small number of institutes and quantitative methods were used in large numbers of institutes, the latex immunoassay (turbidimetric or nepherometric) was predominantly employed. The measuring instruments were various. The upper limit of the reference interval was distributed widely from 5 IU/ml to 40 IU/ml, indicating obvious inter-laboratory differences in RF measurement. Many institutes used the reference intervals recommended by the manufacturers., Conclusion: There are remarkable inter-laboratory differences in RF measurement. Clearly, one of the major issues is the lack of reference materials. We therefore need to establish a reference material of RF, even if it is a temporary one. Scientific societies (Japan College of Rheumatology and Japanese Society of Laboratory Medicine) and manufacturers providing RF measurement reagents should work together for the improvement of RF measurement.

Meditation is a specific consciousness state in which deep relaxation and increased internalized attention coexist. Autonomic nervous activity during meditation was reported to be characterized by decreased sympathetic activity and increased parasympathetic activity. And concerning EEG power, slow alpha power and fast theta power increased during meditation. On the other hand, mental arithmetic (MA) tasks have been reported to induce an increase in blood pressure and heart rate with enhanced sympathetic activity. However, changes in autonomic nervous activity and EEG power during MA tasks and their mutual relationship have not been adequately studied. In the present study, we quantitatively analyzed the changes in autonomic nervous activity using heart rate variability (HRV) and EEG power during MA task in 30 healthy volunteers. MA task induced significant increases in normalized LF, LF/HF ratio (as a sympathetic index), and a decrease in normalized HF (as a parasympathetic index). On the other hand, significant decrease in EEG power (slow theta: 4-6 Hz and fast alpha: 10-13 Hz in the posterior region and fast theta: 6-8 Hz and slow alpha: 8-10 Hz in all the regions) were induced by MA task. Further evaluation of these changes showed a negative correlation between the change rates (with the control resting condition as the baseline) in fast alpha power and that in LF/HF. These results suggest that specific correlated relationships exist between the change in autonomic nervous activity and EEG power depending on the difference in mental task (i.e., meditation or MA).

In Japan, self-monitoring of specific physical conditions is needed, and clinical laboratory testings have been increasingly performed outside of the hospital without the control and/or the supervision of medical doctors. Various items have been measured, which include not only body fat but biochemical markers, cellular analyses for malignancies, sexually transmitted bacterium and so on. Qualitative urinalyses for glucose and protein and for pregnancy, the reagents for which are purchased from pharmacy stores, have been estimated by the examinees themselves. Specimens for most other items described above are collected by examinees themselves, transported via mail or other media and measured in the commercial clinical laboratories. Quality assurance of sampling, transportation and measurement is essential. Therefore, a check-system not only by the owner or manager of the laboratories but also by the non-laboratory staff specialist is needed. Appropriate and exact informations regarding the results must be given to the examinees in consideration of their responsibility concerning the total processes, from preanalytical and analytical to postanalytical processes, of the measurement. Laboratory physicians and technologists should pay more attention to laboratory tests being performed by examinees and outpatients themselves, and participate more in improving and evaluating the usefulness of these tests.

Chronic hypoxia has been newly proposed as a common mechanism of tubulointerstitial fibrosis in the progression of various chronic inflammatory renal diseases, where PAI-1 plays an important role in the accumulation of extracellular matrix (ECM) through inhibition of plasmin-dependent ECM degradation. In the present study, we investigated the presence of PAI-1 in renal tubular cells by immunostaining renal biopsy samples. We also closely examined the effects of hypoxia and TNF-alpha on PAI-1 expression in cultured human proximal renal tubular cells (HRCs). Confluent cells growth-arrested in DMEM for 24h were exposed to hypoxia (1% O2) and/or TNF-alpha at 10 ng/ml for 24 hours. Amounts of PAI-1 protein and mRNA after stimulation were measured by ELISA and TaqMan quantitative PCR, respectively and compared to those in cells incubated under control conditions (18% O2 without TNF-alpha). HIF-1alpha was demonstrated by immunoblot analysis. In crescentic glomerulonephritis, clusters of proximal tubules were specifically stained for PAI-1. Treatment of 24 hours with hypoxia, TNF-alpha and their combination induced a 2.7-fold, a 1.8-fold, and a 4.6-fold increase in PAI-1 protein secretion, and produced a 3.6-fold, a 3.3-fold, and a 12.1-fold increase at the PAI-1 mRNA level, respectively. Immunoblot analysis revealed that hypoxia-inducible factor-1alpha (HIF-1alpha) was markedly accumulated in the nuclear fraction after 16-hours exposure of HPTECs to hypoxia but not to TNF-alpha. In conclusion, hypoxia induces PAI-1 expression via remarkable nuclear accumulation of HIF-1alpha in HRCs. TNF-alpha can enhance this hypoxia-induced PAI-1 expression.

We have recently developed an HPLC method able to separate five lipoproteins (HDL, LDL, IDL, VLDL and chylomicron) followed by cholesterol measurement on each lipoprotein. As an application of this method, this study focused on analyses of triglyceride (TG)-rich lipoproteins, one of risk factor for atherosclerosis. The appearance of midband on electrophoresis is conceivably implicated in atherogenesis. The present study revealed that cholesterol levels in VLDL, IDL and LDL were significantly higher in midband-positive sera than negative sera. Cholesterol levels in remnant-like proteins, another atherogenic indicator, were significantly related to those in VLDL and Chylomicron measured by the present HPLC method. In conclusion, this novel HPLC method can provide valuable information for analyses on TG-rich lipoproteins.

Plasminogen activator inhibitor-1 (PAI-1) may contribute to renal fibrosis because of its involvement in matrix (ECM) accumulation through inhibition of plasmin-dependent ECM degradation. The aim of this study is to determine urinary PAI-1 concentrations and its intrarenal localization in patients with various renal diseases and to identify inducers for PAI-1 expression in human cultured proximal renal tubular cells (HRCs). Urinary PAI-1 concentrations were significantly higher in patients with overt diabetic nephropathy (DN, n=36) than in proliferative glomerulonephritis (PGN, n=8), nephrotic syndrome (NS, n=10) and healthy controls (n=12). Urinary PAI-1 concentrations (ng/gCr) were directly correlated with urinary N-acetyl glucosaminidase (NAG) levels (r=0.58, p<0.05). As for intrarenal localization of PAI-1 antigen, strong stainings for PAI-1 were observed in proximal tubular cells of renal biopsy samples from patients with DN, while no stainings for PAI-1 were found in renal tissues of PGN or NS. Immunoblot analysis revealed the presence of PAI-1 protein in whole cell lyzates from HRCs grown to semiconfluency. Exposure of growth-arrested HRCs with hypoxia (1% O2) or TNF-alpha (10 ng/ml) for 24 hours increased the secretion rate of PAI-1 protein by about 2.0-fold, while 24-hour treatment with high glucose (450 mg/dl) did not increase PAI-1 secretion at all, compared with that of the control cells under normal glucose (100 mg/dl) and normoxia (18% O2). These findings suggest that PAI-1 expression is upregulated especially in the proximal renal tubular cells of DN, which may be explained partially by hypoxia and inflammatory cytokines but not high glucose.

One of the important ethical issues in clinical laboratory medicine is whether organs and/or specimens should belong to the examinees. Tracing back to ancient Greece, an episode of the death of Asklepios, killed by Zeus to revive the dead, and the great contribution of Hippocrates to medicine including the vow and ethics of medicine, have been described. In the relationship between doctors and patients, the former had been superior to the latter for more than 2400 years, however, the situation has been changing from that to the same position since 1960th, along with the development of bioethics from medical ethics. For the promotion of bioethics, world medical associations have contributed declarations and continuous discussion. The declarations are based on the avoidance of actions detrimental to the life, health, privacy or dignity of examinees. On the medical use of human organs and specimens in relation to human rights, the mind and the body, discussion has continued, however, a consensus on the details has not been reached. A view on the use of residual samples for methodological study, teaching and research in the clinical laboratory was proposed by the Japanese Society of Laboratory Medicine in 2002. Briefly, it included confidentiality of the laboratory staff, responsibility of the laboratory director, the absence of a necessity to obtain consent for the use of residual samples for methodological study when they are made anonymous or pooled, and the recommendation to obtain a judgement by an ethics committee for research use. The background and discussion for the proposal and the current situation on how to obtain consent from patients in Japan are mentioned.

Education of clinical laboratory medicine in the medical course of Fukushima Medical University School of Medicine is described. It include lectures (90 min, 15 times) for the 4th grade students and practices for the 5th grade students (one week/2 w, 16 times) and for the 6th grade students (one week/2 w, 4-5 times). In the practice course of 5th grade students, subjects concerning preanalytical issues have been employed to understand the importance. Specimens for practices are taken from every student, which include urines for urinalysis, feces for occult blood test, bloods for Wright-Giemsa staining and cell morphology and coagulation tests, and pharyngeal fluids for Gram staining. In the 6th grade course, residual specimens from anonymous patients are used, and ECG examination is performed by the permission of patients. Through the experiences of sample collections and practices of elementary laboratory tests, students are expected to realize anxiety and pain of patients, to understand the importance of preanalytical variables and to master fundamental laboratory techniques as physicians.

As it has been required to identify pathogenic microbes in shorter times, simple and rapid methods have been developed and used. Here, we summarized the present situation of rapid diagnostic testing in clinical microbiology in Japan, and also presented our results on PBP2' detection. The rapid test kits available in Japan for E. coli, Helicobacter pylori, Salmonella, Streptococcus and Staphylococcus aureus were described. Rapid examination methods are based mainly on immunologic reactions, which included slide agglutination using latex particle, immunochromatography and ELISA. Times required for the identification are 10 to 15 minutes. Moreover, rapid test kits employing PCR are also marketed. Further, we evaluated MRSA-LA "Seiken" which is a rapid detection kit for PBP2' produced by MRSA. The test was shown to be highly sensitive and specific. For the rapid identification of pathogenic microbes, simple and rapid test kits described here will be used more in clinical diagnosis.

Hemodialysis (HD) patients have a high mortality rate due to vascular disease (VD). Therefore, we investigated the effect of uremic dyslipidemia on VD in HD patients, with special consideration of the reverse cholesterol transport (RCT) system including high-density-lipoprotein cholesterol (HDL-C), cholesteryl ester transfer protein (CETP) and its genetic (D442G) mutation. In 414 HD patients, a sub-median HDL-C level (< 48 mg/dl) was an independent risk factor for VD. In the lower HDL-C status, the CETP mutation leading to CETP levels was independently associated with VD. In 210 selected patients, the CETP level was an independent protective factor against VD among those with higher HDL-C levels (> 45 mg/dl). We also measured serum homocysteine (Hcy) levels and examined its association with VD considering that hyperhomocysteinemia is a newly identified risk factor for atheroma. HD Patients (n = 545) had about 3 times the Hcy levels of the general population. A common C677T mutation in the gene of methylenetetrahydrofolate reductase (MTHFR) involved in Hcy metabolism was independently and directly related to serum Hcy levels with TT genotype patients having the highest levels. Patients with the TT genotype were younger and had a shorter duration of dialysis than those with the CT or CC genotype after adjustment for age at the initiation of dialysis, although there was no difference in VD prevalence among the genotypes and no association between Hcy levels and VD prevalence. In conclusion, lower HDL-C and CETP status was a risk factor for VD in HD patients, suggesting the importance of RCT. Serum Hcy levels were markedly increased in HD patients and the TT genotype may be associated with higher mortality. However, a large-scale prospective study is required to clarify whether hyperhomocysteinemia or the TT genotype is a VD risk factor among HD patients.

To know the present situation of rheumatoid factor(RF) measurements, the questionnaire was sent to 82 University or College hospitals in Japan and regional 133 hospitals in Fukushima prefecture. Evaluation of qualitative RF kits commercially purchased in Japan was also performed. In RF measurement, quantitative methods have been increasingly used compared with semiquantitative or qualitative methods. Among quantitative RF measurement methods, latex immunoassay including latex-turbidimetry and latex-nephelometry is most frequently used. The reference values of RF distributed widely ranging from 5 to 40 unit. The 1997 JMA survey still showed interlaboratory differences in quantitative RF measurement. Qualitative RF kits were also shown to have variety of reactivities. To promote the improvement of current situation in RF measurement, "arbitrary" reference materials should be established urgently.

The fecal K concentration and the K-flux activity of erythrocytes were examined to investigate the role of extrarenal K excretion and K transport in peripheral tissue cells in hemodialysis patients. The fecal K concentration of hemodialysis patients was increased significantly(p < 0.01) compared to that of healthy subjects. The K-flux activity of erythrocytes and the serum K concentration correlated negatively(r = -0.482, p < 0.01) in non-diabetic hemodialysis patients, but did not correlate in diabetic hemodialysis patients. K-flux activity and Kt/V for urea correlated significantly(r = 0.412, p < 0.05). The K-flux activity of erythrocytes collected after hemodialysis therapy significantly increased (p < 0.05) compared with that of erythrocytes collected before therapy. We speculated that the K-flux activity of various cells including erythrocytes participates in the regulation of serum K in hemodialysis patients and that the inhibitory factors on K-flux activity are removed partially by hemodialysis therapy.

Monocyte/macrophage (m phi) or mononuclear phagocytic system(MPS) play one of the key roles in defense mechanisms in human as well as animals. Informations as to their biological functions and significance in defense mechanisms or immune systems have increased. However, some of the details are still unknown and/or controversial. Prof. Takahashi lectured on some new findings: development of m phi precedes that of monocyte, factors regulating differentiation of m phi, differences between tissue m phi and monocyte/m phi and so on. He also stressed the complexity of the development and differentiation of monocyte/m phi. Prof Yamamoto lectured on a new factor, S19 ribosomal protein, which enhances migration of monocytes and connects natural immunity and acquired immunity. Following these lectures, all participants realized the complexity and also the mystery of defense mechanisms of our bodies.

It has been reported that serum levels of KL-6 and surfactant protein D(SP-D) can be useful indicators for interstitial pneumonia(IP). In the present study, we evaluated the clinical significance of KL-6 and SP-D by measuring the serum levels of patients with various pulmonary diseases by enzyme-linked immunosorbent assay. Serum levels of KL-6 in patients with idiopathic interstitial pneumonia(IIP), collagen disease with interstitial pneumonia(CDIP), lung cancer(LC) and LC with idiopathic interstitial pneumonia were significantly higher than of those in healthy controls. Moreover, serum levels of KL-6 were significantly higher in patients with active IP than in those with inactive IP. Serum levels of SP-D in patients with IIP and CDIP were significantly higher than of those in healthy controls. When a cut-off level of KL-6 or SP-D in sera was defined as a value of healthy controls representing the means + 2SD, the serum KL-6 positive diagnostic rate for IP(79.2%) was higher than that of SP-D(66.7%). The SP-D positive diagnostic rate for lung diseases other than IP(11.6%) was lower than that of KL-6(34.9%). The serum concentration of KL-6 in patients with the pulmonary diseases significantly correlated with that of SP-D. These findings suggest that KL-6 may be superior in the sensitivity of IP and can be used to evaluate the disease activity of IP. In addition, SP-D may be more specific for IP than KL-6.

Objectives: To determine reference values in several representative serological or immunological examinations in the elderly subjects compared with young adults., Materials and Methods: The group of elderly subjects over 70 years consisted of 630 people living at their own home and 170 people living in nursing homes. The young control group consisted of the 340 university students. ASO and RF were quantitatively measured by latex turbidimetry and FANA by immunofluorescence. Furthermore, several other autoantibodies were also examined by ELISA. The ninety-fifth percentile method was employed for the determination of reference values., Results: The upper limit of reference intervals for ASO was 190 U/ml in the elderly group, while 330 U/ml in the young group. As to RF, it was 25 U/ml (70-79 years) and 37 U/ml (80-89 years), while 15 U/ml in the young. In FANA, x 160 was determined in both the elderly and the young adult subjects. Relatively higher positive frequencies were obtained in the autoantibodies to desmogrein3, dsDNA, cardiolipin and thyroglobulin in the elderly., Conclusion: In the elderly, ASO titers were lower, RF titers were higher, and FANA titers were generally similar to those in young adults. Higher titers were observed for some other autoantibodies.

One of the causes of pseudohyperpotassemia is the induction of potassium (K) release from blood cells by various factors such as cold-storage of blood samples, various drugs including ouabain and so on. We speculated on the effect of storage temperature and therapeutic drugs used on the elevation of K levels in vitro. Dobutamine was selected for in vitro experiment. The therapeutic dose of dobutamine (76 micrograms/l) was mixed with whole blood and incubated at 37 degrees C. A significant increase in K concentration was observed after incubation for 6 hours. These results suggested that many injection drugs used clinically induce K release from blood cells in vitro as well as in vivo and the administration of such drugs could be one of the causes of pseudohyperpotassemia. We developed a simple and practical method to determine the K-flux activity of erythrocytes. Employing this method, hemodialysis patients and aged subjects were evaluated. These were shown to be reduced compared with those of healthy subjects. In 30 hemodialysis patients, K-flux activity and the mean serum K concentration expressed as the average of 4 measurements correlated negatively (r = -0.482, p < 0.01). The K-flux activity of the erythrocytes collected after hemodialysis therapy was increased significantly (p < 0.05) compared with that of erythrocytes collected before therapy. We conclude that the K-flux activity of erythrocytes might participate partly in the regulation of serum K concentration in hemodialysis patients and that the inhibitory factors on K-flux activity might be removed by hemodialysis therapy.