Background and aim: It is well known that the test acceptability for forced vital capacity varies according to the characteristics of examinees and examiners. However, there have been only a few studies investigating the relationship between the acceptability and the characteristics of examinees or examiners in a large number of patients. We investigated the relationship between three acceptability indices for forced vital capacity [extrapolated volume (EV), peak flow time (PEF time), and forced expiratory time (FET)] and the characteristics of patients [age, gender, body mass index (BMI), forced expiratory volume in one second (FEV₁%), vital capacity, number of examinations] or experience of laboratory technologists. Results: Female patients had significantly better EV-index than male patients. Patients with a low BMI had a significantly worse FET-index than those with a high BMI. Patients with FEV₁% < 70% had better indices of EV, PEF-time, and FET than those with FEV₁% ≧ 70%. Less experienced technologists performed better tests in FET-index than well-experienced technicians. Conclusion: The test acceptability for forced vital capacity was different according to the characteristics of patients and experience of laboratory technicians.

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In this study, we examined the effects of β-glucan and lactic acid bacteria, which are components of what are commonly called functional foods. Beta-glucan, lactic acid bacteria, and a mixture of the two were administered orally separately to three groups of BALB/c mice. Each group was then injected with the thyroglobulin antigen, and antibody titers were measured at different stages of immunization. Results showed that antibody production was enhanced at an early stage in mice administered a mixture of β-glucan and lactic acid bacteria as compared with those that received β-glucan or lactic acid bacteria alone. In addition, the mixture induced a significant increase in the phagocytic capacity of macrophages in vitro, for which the synergetic effects of β-glucan and lactic acid bacteria were observed. These results suggest that a mixture of β-glucan and lactic acid bacteria actively stimulates the gut immune system as compared with either component alone, and may be used as an adjuvant for immune antibody production.

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For postmortem diagnosis of diabetes mellitus including fulminant type 1 diabetes, it is necessary to determine the blood glucose concentration before death, and the concentrations of plasma and cerebrospinal fluid 1,5-anhydroglucitol (1,5-AG) and HbA1c were investigated in 59 individuals including diabetic patients (n = 13) and nondiabetic subjects (n = 46) without renal failure on the basis of anamnesis. The postmortem 1,5-AG concentration in the plasma showed linearity with that in the cerebrospinal fluid (r = 0.59). The mean 1,5-AG concentrations in the plasma and cerebrospinal fluid were significantly lower in diabetes mellitus patients than in nondiabetic subjects (plasma 1,5-AG: 6.8 ± 8.3 µg/mL (diabetes mellitus) vs 23.6 ± 13.0 µg/mL (nondiabetes mellitus); cerebrospinal fluid 1,5-AG: 6.1 ± 5.3 µg/mL (diabetes mellitus) vs 19.4 ± 7.9 µg/mL (nondiabetes mellitus)). The plasma and/or cerebrospinal fluid 1,5-AG concentrations in all the diabetic patients were low. Furthermore, the plasma and cerebrospinal fluid 1,5-AG concentrations in nondiabetic subjects with HbA1c 6.5% or more was low. In nondiabetic subjects with normal HbA1c, only the plasma 1,5-AG concentration was low in those who died because of drowning. In conclusion, we consider that measurements of plasma and cerebrospinal fluid 1,5-AG concentrations in addition to HbA1c concentration are useful for the forensic diagnosis of postmortem diabetes mellitus including fulminant type 1 diabetes, and it is better to apply the measurement of cerebrospinal fluid 1,5-AG concentration in those who died because of drowning.

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The reaction principle of the turbidimetric method for urine protein using a cationic detergent (benzethonium chloride) was investigated. The turbidity generation of serum albumin and γ-globulin occurred at approximately pHs 5.5 and 6.0, respectively. This reaction was reversible and dependent on pH, and the generated turbidity was eliminated by decreasing the pH by adding hydrochloric acid. The turbidity generation was considered to be caused by the binding of the positively charged cationic detergent to the negatively charged protein.

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Objective: To study effective restaining techniques for discolored cytological slides Subject and Procedure: Cytological slides, mainly positive cases, from gynecological, respiratory, urinary and body cavity fluid samples, composed of discolored Papanicolaou (PAP) and Giemsa-stained slides were examined. After the removal of coverslips from the PAP-stained slides using xylene, they were processed in various ways, including immersion in phosphate buffer solution or a sodium citrate buffer solution, to analyze the mounting medium removal time, immersion time in a buffer solution, pH and reproducibility. The faded Giemsa-stained slides were immersed in the sodium citrate buffer solution (pH 6.8) before restaining with the Giemsa stain. Result: The immersion in phosphate buffer solution of approximately pH 7.0 for over three hours gave good results in the case of the faded PAP-stained slides. The sodium citrate buffer solution also showed some effect but the most effective was the phosphate buffer solution. The faded Giemsa-stained slides were somewhat improved but their result was not as good as that of the PAP-stained slides owing to the red cells being overstained in blue. Conclusion: The technique that we designed using phosphate buffer solution was considered useful for restaining valuable but discolored slides from previous examinations. We will further study problems including an improved restaining of faded Giemsa-stained slides.

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Ammonium acid urate (AAU) crystal formation in the urine is also seen in children with nephrotic syndrome (NS). For this reason, we investigated the risk factors for AAU crystal formation in children with NS retrospectively. In pediatric nephrology, we confirmed that 85％ of patients presenting with AAU crystal formation were NS patients. The mean age was higher in pediatric NS patients than in general pediatric patients (7.8 ± 5.5 vs 2.2 ± 2.9 years old). Urinary pH was also higher in these patients than in general pediatric patients (6.7 ± 0.3 vs 6.3 ± 0.3). While 25 of 59 (42.4%) general pediatric patients were presented with acute enterocolitis, only 1 of 27 (3.7%) pediatric nephrology patients was presented with acute enterocolitis. Many of the patients examined seemed to have used a steroid, a diuretic drug or an albumin preparation, or have been subjected to infusion therapy. Blood and urinary analyses revealed that low serum total protein level, low serum albumin level, high blood urea nitrogen level, high serum total cholesterol level, high urinary protein-creatinine ratio, and high urinary hyaline cast level were risk factors for AAU crystal formation. Children with proteinuria caused by NS have a higher chance of AAU crystal formation than those without it. AAU crystal formation is linked to the risk of developing postrenal acute renal failure during the recovery phase of NS treatment. Thus, AAU crystal formation is a valuable predictor of the development of postrenal acute renal failure.

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The genotype of the hepatitis B virus (HBV) is an important factor for treatment. Genotyping by EIA has been performed in Japan. In this study, we developed a new system using type-specific polymerase chain reaction (PCR). A total of 84 specimens (EIA genotype A, 8; genotype B, 11; genotype C, 60; genotype D, 1; not typed: 4) were analyzed using the new system. The genotype was determined in 82 of the 84 specimens (97.6%) (genotype A, 8; genotype B, 13; genotype C, 60; genotype D, 1). In addition, it was possible to analyze subgenotypes Aa/Ae and Bj/Ba by tetra-primer ARMS-PCR. Regarding the clinical utility of this new system, it has high capacity to classify the HBV genotypes and subgenotypes, which can support diagnosis.

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The spread of extended-spectrum β-lactamase (ESBL)-producing bacteria in wide areas has become a problem in infection control and infectious disease treatment. In this study, we examined ESBL-producing bacteria isolated in Kagawa University Hospital from 2006 to 2013. Among a total of 434 ESBL-producing isolates, 372 (85.7%) were Escherichia coli, 49 (11.3%) were Klebsiella pneumoniae and 13 (3.0%) were others species. The number of ESBL-producing E. coli increased from 15 strains in 2006 to 84 strains in 2013 and that of ESBL-producing K. pneumoniae from 2 to 19 strains. The detection rates of ESBL-producing isolates gradually increased and reached 20.0% and 9.0% for E. coli and K. pneumoniae, respectively, in 2013. For E. coli, the most commonly detected genotype belonged to the CTX-M-9 group (145/309 strains; 46.9%). In addition, the detection rate of the CTX-M-1 group tended to increase. For K. pneumoniae, the CTX-M-9 group (61/81 strains; 75.3%) was the most, prevalent, suggesting the spread of a single genotype by nosocomial transmissions. It is considered that active fecal surveillance and appropriate infection control are necessary for preventing the spread of ESBL-producing bacteria in the future.

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The patient was an 8-year-old girl who developed high-grade fever and examined by a primary care doctor. High C-reactive protein level (CRP, 14.5 mg/dL) and white blood cell count (WBC, 18,900/μL) in a blood test were observed. She was administered an intravenous antibiotic called ceftriaxone (CTRX; total dose of 3 g/kg body weight). An elevated WBC in urine indicated urinary tract infection; thus, she was subjected to intensive examination and treatment. On admission, a contrast-enhanced computed tomography revealed a poorly enhanced area in the bilateral kidneys. Ultrasonography (US) revealed left hydronephrosis with ureteropelvic wall thickening. The diagnosis was acute focal bacterial nephritis for which she was treated with a panipenem/betamipron after administration of 1 g of CTRX (CTRX total dose of 4 g/kg body weight for 2 days at this point). Two days later, US revealed a focal hyperechoic area with poorly defined margins and a hypovascular area by color Doppler imaging. US showed no stones in the gall bladder on admission. However, on hospital day 3, US revealed sandlike and floating hyperechoic foci in the gall bladder, that is, a biliary sludge was formed. The patient had no abdominal symptoms. The follow-up US, which was performed on days 5 and 7, revealed that the size of the biliary sludge gradually decreased. Eventually, on day 11, US showed that the biliary sludge had disappeared. We encountered a patient with CTRX-associated pseudolithiasis, and US was helpful in demonstrating the appearance and disappearance of a biliary sludge. Additionally, it is important that we determine whether a primary doctor used CTRX as part of the patient’s medication history.

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A general examination of cerebrospinal fluid has a high clinical significance in the diagnosis of central nervous system diseases. Furthermore, it is a very important examination for the confirmation of a neoplastic disease of the central nervous system, the determination of a treatment strategy, and the observation of recurrence and progress. The observation of cells in the cerebrospinal fluid is one of the most important aspects of the examination. Cell observation by a general examination of cerebrospinal fluid can be carried out promptly without fixation and complicated staining procedures. Because a general clinical technologist grade system has been established, the cell observation technology for puncture liquid as well urine sediment has improved. The certification of general clinical technologists has to be advanced to the next stage. I report on four cases wherein information about the disseminated transition of cerebrospinal fluid was promptly submitted to the clinical side.

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Monoclonal immunoglobulins (M-proteins) have various effects on measurement results. We encountered a case in which a patient serum showed an abnormal reaction time course in the AST assay. In this case, we proved that the presence of the IgM-κ-type M-protein in the patient serum results in turbidity in the reaction of the patient serum with the first reagent in the AST assay. We found no abnormal reaction in comparative studies using eight other reagents from different manufacturers, such as reagents of the JSCC transferable method. Then, we focused on the buffer solution of the reagent. We confirmed the reaction of the patient serum with three types of buffer solution. We therefore concluded that the abnormal reaction of the present case was due to the turbidity caused by the low ionic strength of the reagent in the AST assay.

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Mitochondrial encephalomyopathies include various symptoms such as central nervous system damage, muscle strength loss and diabetes. Leigh syndrome, which primarily affects children, is one of the mitochondrial encephalomyopathies classified on the basis of presenting clinical conditions and laboratory findings. In this study, we report the association between evoked potentials and clinical conditions in Leigh syndrome patients. The study subjects included five patients diagnosed as having Leigh syndrome. We recorded the short-latency somatosensory evoked potential, auditory brainstem response and visual evoked potential of all the patients. All the patients aged <1 year showed early symptoms of poor sucking or eye nystagmus; with time, all the patients regressed, but their symptoms worsened. We found no specific abnormality pattern for any patient by comparing the evoked potentials and clinical conditions. However, the patients with N20 and N30 scalp waves showed relatively better early clinical manifestation than those who lost scalp waves owing to gradual disease progression after regression. This observation can be attributed to the functions of the primary sensor area of N20 origin, basal ganglia of N30 origin, or early development of the subcortical pathway. The cumulative findings may indicate an association between the existence of scalp waves and early clinical manifestations. Thus, the scalp wave may be used as a reference indicator for predicting Leigh syndrome prognosis.

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Mediace TPLA (Kyokuto Pharmaceutical Industrial Co., Ltd) has been used at our hospital as a diagnostic reagent for syphilis by the latex agglutination method. However, we often encountered high false-positivity rates owing to the influences of chyle and hemolysis. In this study, we evaluated anti-Treponema pallidum antibody reagents based on recombinant antigens from three manufacturers (Fujirebio Inc., Denka Seiken Co., Ltd., and Shino-Test Corporation). Good results were obtained in terms of reproducibility, prozone test, and other types of test. High sensitivity and specificity were obtained; in particular, the specificity was comparable to that obtained by the FTA-ABS method. It is expected that the required frequency of confirmatory testing by the TP antigen neutralization test and other methods will be reduced with the use of the anti-T. pallidum antibody reagents based on recombinant antigens in place of those based on antigens prepared from purified bacterial cells.

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In medical practice, misidentification of patients is a serious problem. It is possible that a reverse in steps will occur after fixation, especially in the washing of organs, photography and dissection. However, when identification tags are directly attached to materials, the number of accidents decreases. Here, we present a new identification method using “Bano’k-gun” and “Tag-pin”, which are used as price tags in the clothing industry. A trial involving 42 cases of surgical materials and authentication of matching surgery patients to pathological specimens after resection was carried out. This technique was 100% successful in terms of both recognition rate and labeling index. The handling procedure was simple and coordination with the digital data was smooth. Washing can be carried out separately, and needles can also be used for infected materials. Tissue damage around the “pin” is negligible and does not affect the diagnosis. We believe that this method is feasible and safe for preventing misidentification. Most of the misidentifications of pathological specimens are caused by human error. We propose a method of preventing such misidentifications.

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Good results were obtained using sputum specimens and Grocott’s staining of bronchoalveolar lavage specimens using the BD SurePath^TM liquid-based cytology system. The system removed mucus and blood from all the specimens, and staining was carried out with a smear of 13 mm circumference through ionic bonds. Cell abrasion was improved, and good Grocott’s staining was possible.

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Heart-type fatty acid-binding protein (H-FABP) is an early marker of acute coronary syndrome (ACS). Immunochromatography is a widely used method for the measurement of H-FABP. However, this method is restricted by individual differences owing to visual judgment, and only a qualitative judgment is possible. Therefore, we evaluated the reagent from the LATEX H-FABP KIT (YAMASA) for the quantitative measurement of H-FABP. This reagent had a favorable within-run precision coefficient of variation (CV) in the range of 0.65–3.98% and between-day precision CV in the range of 1.28–5.57%. Additionally, coexisting materials had no effect on the reaction. In ACS patients, the sensitivity and specificity were 72% and 59%, respectively. However, it was considered desirable to consider the time for obtaining a blood sample from a heart attack victim.

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A specific medical examination in Kumamoto Prefecture demonstrated that the rates of examinees with HbA1c levels above the reference level (5.6%) significantly varied among regions. To find the cause and take corrective action, differences in the measurement results due to instruments and reagents were investigated. Blood samples were taken from 20 patients from whom informed consent was obtained. JCCRM411-2 was used as a reference standard. HbA1c level was measured for each sample using HPLC, turbidimetric immunoassay, and an enzymatic method. The measurements demonstrated a maximum difference of 0.3% in blood samples from patients with a HbA1c level of 5.6% among the methods. The measurements were 0.2 and 0.1% higher by the HPLC and enzymatic methods than that of the reference standard, respectively, and closely corresponded to those by the turbidimetric immunoassay. Different instruments and reagents may have caused the differences in measured HbA1c levels. To solve this problem, efforts should be immediately made to standardize the HbA1c measurement methods.

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We performed basic studies of the reagent ‘auto ASO・BML’ for antistreptolysin O (ASO) measurement based on the latex turbidmetric immunoassay. This reagent showed CVs of within-run precision of less than 1.0% and CVs of between-day precision of less than 2.0%. The detection limit was 8.0 IU/mL and the dilution linearity was also good. No effects from interfering substances were detected. In addition, the correlation with ‘LPIA ACE ASO Ⅱ’ of the comparison product, the measurement principle of which is similar to that of this reagent, was good (y = 0.939x – 0.9, r = 0.991, n = 155). ‘Auto ASO・BML’ showed sufficient reagent performance in routine tests.

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Here, we report on a study of acid-fast bacilli detection that we conducted at our clinic. We examined the detection of acid-fast bacilli, age distribution, sex differences, and the positivity rates for different test items in 1,898 specimens that were tested for acid-fast bacilli during a six-year period between April 2006 and March 2012. Over these six years, 12.0% of the specimens tested positive for acid-fast bacilli. When the age distribution was examined, no major changes were observed in the rates of detection of tuberculous or nontuberculous mycobacteria among patients aged 70 years or older. When sex differences were examined, the rate of detection of tuberculous mycobacteria was found to be high among men initially, but slowly decreased over time and became 50.0% for both men and women in 2010. The rate of detection of nontuberculous mycobacteria was high among women in 2006, but decreased afterward. Although clear differences between the sexes (for example, tuberculosis is predominant among men whereas nontuberculous mycobacterial infection is predominant among women) have been found, this study did not show such a trend. The patient age was 70 years or older in more than half of the cases where tuberculous or nontuberculous mycobacteria were detected. We sometimes also see young patients infected with tuberculosis bacilli at our clinic; thus, it is necessary to proactively consider the possibility of tuberculosis in young patients as well as elderly patients. When compared by test item, culture tests showed that a high percentage (≥ 80%) of specimens were positive for both tuberculous and nontuberculous mycobacteria. However, bacteria can sometimes be detected by smearing or Polymerase Chain Reaction (PCR) even when a specimen culture shows negative results, which indicates that it is important to conduct an assortment of tests. In addition, it is important to manage infections quickly and appropriately to prevent their spread after considering the geographical distribution.

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Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph⁺ALL) develops owing to the formation of BCR-ABL tyrosine kinase due to a reciprocal translocation between chromosomes 9 and 22. Although Ph⁺ALL treated by conventional chemotherapy reportedly has a poor prognosis, imatinib mesylate has been shown to markedly improve both complete remission (CR) rates and overall survival rates. Therefore, we retrospectively investigated the outcome of chemotherapy combined with a tyrosine kinase inhibitor (TKI) in 5 patients with newly diagnosed Ph⁺ALL (median age, 46 years; 2 males and 3 females) treated in the past decade. According to the results, the CR rate was 100% and 3 of the 5 patients survived without progression of the disease. TKI showed remarkable efficacy in these patients. A majority of the Ph⁺ALL patients achieved CR during the TKI treatment, but some patients relapsed with the emergence of imatinib resistance. Therefore, we should select the best TKI among several available drugs, and establish the best combination of TKI and stem cell transplantation to improve the clinical outcome of Ph⁺ALL. It is important to simultaneously monitor BCR-ABL chimera while analyzing the ABL gene mutation.

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