Kazunari Kumasaka

Frequency of intron 14 splicing defect of cholesteryl ester transfer protein gene in the Japanese general population — relation between the mutation and hyperalphalipoproteinemia

Cholesteryl ester transfer protein (CETP) deficiency, which has been found only in Japan, is characterized by marked hyperalphalipoproteinemia (HALP) and abnormalities of both low density and high density lipoproteins. We have reported that this deficiency is commonly associated with a G-->A mutation at the intron 14 splice donor site of the CETP gene (Yamashita et al., Biochem. Biophys. Res. Commun., 170 (1990) 1346-1351). In the current study, we determined the frequency of this mutation in Japanese subjects by using polymerase chain reaction. A single primer-template mismatch of one base pair from the CETP gene mutation permitted the introduction of a cleavage site for Nde I in mutant alleles but not in normal ones. Out of 171 patients with marked HALP whose serum HDL-cholesterol was more than 100 mg/dl, 6 (3.5%) subjects were homozygous and 48 (28.1%) were heterozygous for this mutation. Furthermore, in unrelated 512 healthy Japanese subjects, 5 (0.98%) were identified as heterozygotes. Relative allelic frequency of A at the intron 14 splice donor site was 0.0049 and the frequency of homozygous CETP deficiency was estimated to be approximately 1/42,000. These results demonstrate that this common mutation may be frequent in the Japanese population. Although HALP is very heterogenous, this mutation could be one of the major causes of marked HALP.

Body mass index variations by age and sex, and prevalence of overweight in Japanese adults

OBJECTIVE: To determine the reference value of BMI for Japanese subjects and to estimate the prevalence of overweight based on this reference value. DESIGN: Epidemiological analysis with the LMS method, which provides a way of obtaining normalized BMI distributions. SUBJECTS: 7508 Japanese subjects aged 18–69 y in 1993. MEASUREMENTS: Height, age and body weight. CALCULATION: BMI was calculated and tables for percentiles of BMI were plotted against age and sex. Furthermore, the prevalence of overweight was estimated based on 85th percentile of BMI in the men and women 20–29 y of age, who were considered the reference group. RESULTS: The geometric mean BMI and the prevalence of overweight in men was highest in the 30–39 y age group. For women the maximum BMI and prevalence of overweight occurred in the decade 50–59 y. The cut-off points for overweight in this sample were 24.7 kg/m2 for men and 22.6 kg/m2 for women. These are considerably lower than the figures of 27.8 kg/m2 and 27.3 kg/m2 estimated for Americans. CONCLUSIONS: The prevalence of obesity in Japanese populations should be estimated using ethnic specific values of BMI, rather than those drawn from Caucasians who tend to have higher BMI in each age group. The prevalence of overweight is increased as age increased in both sexes, especially in women.

Nephrotic syndrome with portal, splenic and renal vein thrombosis. A case report.

Background: Thromboembolism is known as a major complication of nephrotic syndrome, but only 4 cases of portal vein thrombosis have been reported as a complication of nephrotic syndrome. All of these 4 cases had acute symptoms, and 3 of 4 were in relapsing phase of nephrotic syndrome when thrombi were found. We describe here a case of 51-year-old woman with fresh nephrotic syndrome that was asymptomatically complicated by portal, splenic and renal vein thrombosis. Conclusion: In the presence of fresh nephrotic syndrome of minimal change, asymptomatic and widely distributed, including portal vein, thrombus formation occurred. If the clinical course shows resistance to therapy, we must consider the complication of venous thrombosis. Anticoagulant therapy with heparin and warfarin was effective and all thrombi disappeared without any other complications.

Development and performance of an enzyme immunoassay to detect creatine kinase isoenzyme MB activity using anti-mitochondrial creatine kinase monoclonal antibodies

Abstract Objective: The MB fraction of creatine kinase (CK-MB) has long been used as a cardiac marker. It is known that the CK-MB immunoinhibition method lacks selectivity and accuracy, because the appearance of macro CK type 2, corresponding to mitochondrial creatine kinase (MtCK) in some patient serum may render CK-MB activity measured by conventional method abnormally high. Thus, to improve the specificity and accuracy of the CK-MB assay, we developed two types of monoclonal anti-MtCK antibodies against sarcomeric MtCK and ubiquitous MtCK, and present herein the performance of a new method using these antibodies. Material and methods: The performance of our test for detecting CK-MB activity was compared with other methods, and the range of CK-MB activities in normal human serum was investigated. Results: The two types of monoclonal antibodies developed by us were isoenzyme-specific to sMtCK or uMtCK. The correlation coefficients of our method and conventional method to electrophoresis were 0.973 and 0.873, respectively. The mean CK-MB activity in normal human serum by our method and the conventional method was 2.4 and 11.7 U/L, respectively. Thus, our data indicated that about 80% of CK-MB activity, determined using the conventional method, seems to correspond to the MtCK activity. Conclusion: Our method is novel in offering higher accuracy of measuring true CK-MB contents in human serum as compared to the conventional method. The possibility of accurately estimating CK-MB activity by our method which can inhibit MtCKs in healthy person and patient serum is likely to bring a break-through in clinical diagnostics.

A case of Benign familial hyperphosphatasemia of intestinal origin

We recently encountered a case of hyperphosphatasemia, in which > 90% of serum alkaline phosphatase (ALP) was of intestinal origin. The patient, a 51-year-old man, was found to have hyperphosphatasemia (2,341 U/L) during a routine medical check-up. All other laboratory tests and physical findings were normal. The agarose gel electrophoresis pattern of the patients serum ALP was identical to that of common intestinal ALP from healthy adults, and only a single band of intestinal ALP was detected by immunoaffinity electrophoresis. In addition, 89% of total ALP was defined as intestinal ALP by an immunoprecipitation method. The molecular mass of the ALP was 154 kDa, almost identical with that of adult duodenal ALP. Analysis of the sugar chain structure showed an increased la fraction (74%) compared with adult duodenal ALP. Genealogical study revealed that two persons in the 5 members of the probands family had hyperphosphatasemia of intestinal origin, indicating possible autosomal dominant inheritance.

Peritonitis Associated With Pasteurella multocida: Molecular Evidence of Zoonotic Etiology

A patient on continuous cyclic peritoneal dialysis for chronic kidney disease due to type 2 diabetes mellitus developed peritoneal dialysis‐associated peritonitis induced by Pasteurella multocida that was isolated from a sample of dialysis effluent. The route of infection was unknown for this case; however, P. multocida was also isolated from a culture of a pharyngeal swab obtained from the patients cat. There was no evidence that the cat had bitten and ruptured the peritoneal dialysis tubing or bags. Pulsed‐field gel electrophoresis (PFGE) showed that the P. multocida isolated from the patient was completely identical to the strain isolated from the domestic cat. As there is a rise in the pet‐keeping population, an increase in zoonoses is to be expected. It is necessary to be carefully informed of hygiene rules in keeping pets because a pet may transmit zoonoses, even on casual contact.

Abnormal alkaline phosphatase of hepatic type in cerebrospinal fluid of a patient with intracranial metastasis from lung cancer.

High alkaline phosphatase (ALP) activity was found in the cerebrospinal fluid of a patient with intracranial metastases from adenocarcinoma of the lung. On agarose gel electrophoresis of the major ALP isoenzyme found in the cerebrospinal fluid, its mobility was different from those of the usual serum ALP isoenzymes. This abnormal mobility might be due to the linked glycan phosphatidylinositol anchor in the ALP molecule, as the mobility became the same as that of the common liver type ALP after treatment with phosphatidylinositol specific phospholipase. The immunochemical antigenicity of the cerebrospinal fluid ALP was identical with that of the common serum liver type ALP, but its sugar moiety was similar to the membranous liver-type ALP rather than the serum liver type ALP. The molecular size of the cerebrospinal fluid ALP was 140 kilodaltons, 12 less than the common serum liver type ALP, suggesting that the ALP in the patients cerebrospinal fluid was derived from the intracranial metastatic carcinoma.

Relationship between oligomer and functional serum mannose-binding lectin in chronic renal failure

Eur J Clin Invest 2010; 40 (10): 865–873

Circannual Rhythm of Laboratory Test Parameters among Chronic Haemodialysis Patients

Background/Aims: Seasonal variations in laboratory test results have been pointed out in dialysis patients. Although the mechanism for this phenomenon is not clear, this could result in changes in dialysis and medication prescriptions. We investigated the effect of the circannual rhythm on laboratory test parameters in chronic haemodialysis patients. Methods: Data of 38 laboratory test parameters were collected every month and analyzed for 150 stable haemodialysis patients, with non-linear sine wave regression and paired t test between data of peak and trough months. Results: Serum urea nitrogen, unsaturated iron binding capacity, lactate dehydrogenase, alkaline phosphatase, amylase, and neutrophil count showed significant circannual rhythms with high amplitudes. Additionally, serum creatinine, uric acid, chloride, calcium, phosphate, magnesium, total cholesterol, total protein, leucocyte count, mean corpuscular haemoglobin level, mean corpuscular haemoglobin concentration, and platelet count showed significant circannual rhythms with little amplitudes. Conclusions: The circannual rhythm of laboratory test parameters could be attributed to seasonal variations in food intake. Awareness of these variations should be taken into account in the interpretation of laboratory results.

Evaluation of Serological Diagnosis Tests for Tuberculosis in Hemodialysis Patients

Abstract:  Patients receiving hemodialysis are generally considered to be at increased risk of developing tuberculosis. In the current study, in order to evaluate the usefulness of serological tests in dialysis patients, serum antibodies for tuberculous glycolipids antigen (TBGL) and for lipoarabinomannan (LAM) were measured in hemodialysis patients. The present study included 243 hemodialysis patients. Serum antibodies for TBGL and LAM were measured. Tuberculin skin tests were carried out and chest X‐rays evaluated at the same time. There were no patients with active tuberculosis at the time of blood sampling. Thirty‐six patients (14.8%) and 25 patients (10.3%) were positive for anti‐TBGL antibody and anti‐LAM antibody, respectively. One hundred and fifty‐five patients (63.8%) were positive for tuberculin skin testing and 123 patients (50.6%) had old pulmonary tuberculosis on their chest X‐ray. There was no significant correlation between the results of anti‐TBGL antibody and anti‐LAM antibody. There were no relationships among the results of tuberculin skin test and the two serological tests. However, positivity of anti‐TBGL antibody and anti‐LAM antibody was significantly higher in patients with findings of old tuberculosis on the chest X‐ray than those without findings. The current results show that these serological tests are positive more frequently in hemodialysis patients without any proof of active tuberculosis than in healthy subjects (2%) and careful interpretation is necessary for relevant results.