We found and identified five heterozygous dysfibrinogenemias with γN308K (AAT→AAG) mutation in three families by coagulation screening tests and direct sequence analysis of PCR-amplified DNA fragments. Three dysfibrinogens were designated as fibrinogens Matsumoto II, Matsumoto XI, and Nagano I. The patients’ fibrinogens purified from plasma using immunoaffinity-chromatography were subjected to thrombin-catalyzed fibrin polymerization. In the absence of Ca²⁺, the patients’ fibrinogens showed impaired thrombin-catalyzed fibrin polymerization in comparison with normal control fibrinogen. On the other hand, in the presence of 1.0 mM Ca²⁺, the polymerization of all the patients’ fibrinogen was improved as compared with those in the absence of Ca²⁺. Although Single Nucleotide Polymorphism (SNPs) analysis showed that all patients might be derived from a single proband of mutation, the degree of impairment of polymerization was markedly different among affected patient’s fibrinogens. Moreover, SDS-PAGE analysis indicated that the ratio of the amount of the mutant γ-chain to that of the normal γ-chain was not significantly different among the affected patients’ fibrinogens. In conclusion, we speculate that the increase in the amount of heterodimers consisting of normal and mutant γ-chains (namely, the decrease in the amount of homodimers consisting of the normal γ-chain) leads to the marked impairment of thrombin-catalyzed fibrin polymerization.

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Purpose: The purpose of this study is to investigate whether the wireless 12ch ECG system can reduce the required number of operators and time compared with the traditional cabled wired 12ch ECG system during exercise stress echocardiography.
Subjects and Methods: Thirty-one consecutive subjects (24 males; mean age, 66 ±11 years) who were suspected of having angina pectoris and planned to undergo exercise stress echocardiography were enrolled in this study. The stress echocardiography using the wireless 12ch ECG system was performed by two operators. Meanwhile, the stress echocardiography using the cable wired 12ch ECG system was carried out by two strategies, i.e., one performed by three operators (Method A) and the other by two operators (Method B). The elapsed time from the end of exercise to acquiring echo images was compared among three methods.
Results: All of the stress echocardiographic studies using the wireless 12ch ECG system (n = 12, 10 males; mean age, 67 ± 8 years), cable wired 12ch ECG system by three operators (Method A) (n = 12, 9 males; mean age, 64 ± 14 years), and cable wired 12ch ECG system by two operators (Method B) (n = 7, 5 males; mean age, 68 ± 12 years) were performed without any compilations. The elapsed time for the with wireless 12ch ECG system was 19 ± 3 seconds and that for Method A was 18 ± 4 seconds (p = N.S.), which shows that the wireless 12ch ECG system is as useful as the standard cabled wired method even when performed by two operators. However, it took 31 ± 9 seconds for Method B, which is longer than the times for the other two methods.
Conclusion: The wireless 12ch ECG System might reduce the operator’s burden and make the exercise stress echocardiography easier, implying that it is a valuable tool during exercise stress echocardiography.

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We investigated the isolation rates of the four species of ESBL-producing intestinal bacteria isolated from urine and blood in four medium-size hospitals in fiscal years 2007, 2009, and 2011. Regarding the transition of the number of ESBL-producing isolates from both urine and blood, the number increased with each passing year in both outpatients and inpatients. The ESBL-producing bacteria with the highest isolation ratios are Proteus mirabilis, Escherichia coli, Klebsiella oxytoca, and Klebsiella pneumoniae, in this order.
Significant differences in the isolation rates of ESBL-producing bacteria from inpatients among facilities were observed. These results suggest that active fecal surveillance on admission for patients with risk factors would be useful for hospital infection control.

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Background and aim: Ultrasonography of an inguinal mass is not common in clinical practice; so the aim of this study is to determine its clinical significance. Materials and methods: Two hundred and fifteen patients with an inguinal mass and who underwent ultrasonography between June 2010 and May 2012 were involved in the study. The monthly number of ultrasonographies of an inguinal mass, the ultrasonographic diagnosis, in comparison with the diagnosis made by palpation, and accuracy of the diagnosis of the type of groin hernia were evaluated. Results: The monthly number of ultrasonographies of an inguinal mass increased to 17, which was higher than the number of operations for an inguinal mass. The diagnoses included groin hernia, spermatic (Nuck) hydrocele, lymph node, round ligament varicosity, and ectopic endometriosis. The ultrasonographic diagnosis was different from the diagnosis made by palpation in 13 cases (6.0%). Comparison of the ultrasonographic diagnosis with the final diagnosis made by the operative findings in 153 cases revealed that the diagnostic accuracy for the type of groin hernia was 82%. Conclusion: Ultrasonography is useful for the diagnosis and treatment of an inguinal mass.

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In this report, we discuss our analytical and clinical evaluations of assays of HCC tumor markers, namely, AFP, AFP-L3, and PIVKA II, using a fully automated microfluidic immunoassay system with the laser-induced fluorescence (LIF) detector, the μ TAS Wako i30 (Wako Pure Chemical Industries, Ltd., Osaka, Japan). The robust performance of these assays on μ TAS Wako i30 satisfied the requirements for daily clinical use, and the combination of AFP-L3%, PIVKA II, and total AFP measurements resulted in higher true positive and true negative rates for HCC diagnostics than AFP measurement alone. These observations suggest that the μ TAS Wako i30 AFP-L3% and PIVKA II assay combination is a useful tool for aiding in the diagnosis of HCC.

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Campylobacter jejuni usually hydrolyzes hippuric acid to benzoic acid and glycine, and a simple hippurate hydrolysis test is used to rapidly distinguish C. jejuni from C. coli in many clinical microbiological laboratories. However, sometimes the test gives variable results, which might be due to the inadequate number of bacterial cells in the test sample. In this study, we investigated the optimum number of bacterial cells required to perform the test smoothly. A total of 100 Campylobacter strains were included in this study, and test bacterial cell suspensions were prepared matching the McFarland units 0.5, 1, 2, 3, 4, and 5. Ninhydrin was added to the cell suspension and the color change was measured at an optical density of 540 nm. The test of bacterial suspensions of C. jejuni matching the McFarland units 1, 2, 3, 4, and 5 showed positive results for all suspensions. On the other hand, bacterial cell suspension of C. coli matching a McFarland unit of more than 5 gave false positive results. Reproducible results were obtained with a bacterial suspension that matched the McFarland unit 2 and this number of bacterial cells may be used for the hippurate hydrolysis test to distinguish C. jejuni from C. coli.

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In this study, we aimed to explore the relationship of HLA allele frequency with the primary disease of IgA, NIDDN, and CGN in 354 renal transplant recipients.
The HLA allele genotype was measured by the PCR-rSSO technique. The frequency of DRB1*04:05 in the IgAN group was significantly higher than that in the donor control group (23.611% vs. 15.395%, odds ratio (OR) = 1.699, 95% confidence interval (CI): 0.404–0.856, p = 0.010). The frequency of DRB1*04:05 in the NIDDN group was significantly lower than that in the donor control group (9.615% vs. 15.395%, OR = 0.585, 95% CI: 1.033–2.831, p = 0.040).
The frequency of DRB1*04:05 in the CGN group was significantly higher than that in the donor control group (22.1873% vs. 15.395%, OR = 0.638, 95% CI: 1.107–2.215, p = 0.010).
We conclude that our study suggests an association between HLA allele frequency and primary disease in renal allograft recipients.

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Cytological preparations of cervical cancer samples collected after radiotherapy may be dry owing to vaginal dryness, with which the detailed observation of cells is difficult. We processed cytological specimens of 12 patients after radiotherapy for cervical cancer using two procedures and compared the cytological findings: samples were processed employing the standard alcohol fixation method in one procedure, and dried and rehydrated before fixation in the other procedure. About one-half and more than 75% of cells were dried in 4 and 1 standard alcohol-fixed preparations, respectively, but more than 75% of cells could be observed favorably in all dried-and-rehydrated preparations, confirming the usefulness of the drying-and-rehydration method. The drying-and-rehydration method may be useful for preparing samples when dryness after radiotherapy is expected.

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The sulfosalicylic acid method is used to measure urine protein concentration. The relationship between the turbidity/precipitated protein concentration ratio and the pH of the reaction mixture in this turbidimetry was examined. The ratio decreased with increasing pH of the reaction mixture. This finding indicates that the radius of the particle precipitated by sulfosalicylic acid decreases with increasing the pH and that urinary pH affects turbidity generation and causes variations in measurements.

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The patient was a 22-year-old female diagnosed as having acute myeloid leukemia. The ABO blood typing test using anti-A showed mixed field agglutination by both the tube test and the column agglutination technology on admission. The transferase activity of A antigen was stable. The A antigen activity decreased after the start of remission-induction therapy, and disappeared on the 11^th day after admission. She was transfused with two units of group A, D-positive red blood cells (RBCs) on the same day, and the A antigen activity increased to (4+) (256 times) on the 14^th day. Interestingly, in this case, at the time when the A antigen activity of RBCs decreased in the body, the A antigen activity also decreased simultaneously in blood samples kept in cold storage. However, the A antigen activity did not decrease even in cold storage after reactivation. The cause of the decrease in ABO antigen activity in leukemia still remains incompletely clarified. In this case, the decrease in A antigen activity is considered to be closely associated with the weakness of A antigen structure because the A antigen activity of RBCs in the body and cold storage decreased at the same time.

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This is a case report of an 86-year-old male with pneumonia caused by Pasteurella multocida infection. The patient had hypertrophic cardiomyopathy and was on postoperative hormone therapy for bladder cancer. The patient presented with a four-day history of productive cough and nocturnal wheeze, and was diagnosed as having pneumonia during a routine outpatient visit. Following admission, treatment with sulbactam/ampicillin was initiated, but was later changed to Ceftriaxone as the sputum Gram staining showed small Gram-negative rods leading to suspicion of infection by Haemophilus species. His condition improved rapidly and the patient was discharged after eight days. The bacterium seen on Gram staining was later identified by culture as Pasteurella multocida, and the occurrence of pneumonia caused by this organism was consistent with the fact that the patient had intimate daily contact with his pet dog. On Gram staining, Pasteurella species is often mistaken as a Haemophilus species owing to their resemblance in appearance. Infection due to a Pasteurella species should be suspected when careful history-taking reveals intimate contact with animals. Besides the well-known association with animal bites, increasingly more cases of respiratory infections with Pasteurella species secondary to intimate contact with pets are being reported, as in our case, and intimate contact with pets should be recognized as an important mode of transmission of these organisms.

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We have encountered an example of a case of autoimmune pancreatitis (AIP) accompanied by lower bile duct contraction in a male patient who was operated on the basis of cytological and radiological findings. Here we report bibliographic considerations in relation to this case. A 54-year-old male with jaundice was introduced to our hospital by a general practitioner. In a blood test, the bilirubin and biliary enzyme levels were high. Since the extension of the internal and external bile duct of the liver was detected by CT, he was diagnosed as having obstructive jaundice and hospitalized for detailed examination and medical treatment. High levels of the tumor markers CEA and CA19-9 were observed in a blood test at the time of hospitalization. Magnetic resonance cholangiopancreatography and ultrasonography were performed, which revealed a lower bile duct tumor and he was diagnosed as having obstructive jaundice. A cytological examination of the bile, led to a suspicion of malignancy and the head of the pancreas duodenum was excised. A cytological diagnosis of the excised duodenum revealed a mammillary cluster, which overlapped with the suspicion of neoplastic lesion. Macroscopic analysis showed no tuberosity lesion in the pancreas, but the histopathological diagnosis was lymphoplasmacytic sclerosing pancreatitis (LPSP) and cholangitis. Blood plasma cell analysis revealed IgG4 positivity in about 50 cells in the high power field. He was diagnosed as having AIP accompanied by IgG4-related sclerosing cholangitis (IgG4-SC). AIP is an inflammatory disorder for which steroids are effective. At present, It is not clear whether it becomes a risk factor for development of malignant tumor symptoms or whether an operation is necessary or not. There is also no standardization of cytological characteristics and it is necessary to collect cases from now on and to explore the feature of the cell that appears in bile.

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Exercise-induced vasospastic angina (EIVA) is a relatively rare disease reported to be recognized in about 40% of vasospastic angina. EIVA must be differentiated from common effort angina irrelevant to circardian rhythm. We report the case of a 61-year-old male with an angina episode with ST segment elevation in ECG during a treadmill exercise test. A significant vasospasm of the right coronary artery was demonstrated by coronary angiography and intracoronary injection of methylergometrine maleate. These findings suggest that the expected mechanism of EIVA might be ascribed to increased sympathetic tone in the morning closely associated with circadian variation, endothelial vascular dysfunction in the epicardial coronary artery, and smoking. Moreover, determination of when EIVA occured may be very supportive in the diagnosis of EIVA. As a result, recalling EIVA may lead to a reduction in unfavorable complications when the exercise stress test is carried out to diagnose EIVA.

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Trichophyton tonsurans is a pathogenic dermatophytic fungus causing tinea capitis. It has been identified among judo and wrestling athletes in Japan since 2000. In this study, we isolated a dermatophytic fungus from the head pus of an 8-year-old male who practices judo. It was classified as belonging the genus Trichophyton by observation of its slide culture under a microscope. The ITS and D1/D2 regions, which are encoded by the rRNA gene, were amplified by PCR and then sequenced to identify the Trichophyton species. The fungus isolated from this case was identified as T. tonsurans because it showed high homology to this species (ITS, 98.3%; D1/D2, 100%) in a BLAST search using the DDBJ database (DNA Data Bank of Japan). The suggested identification method improved the credibility of results by employing simultaneous confirmation of homologies from two or more independent regions of the gene. Hence, it may contribute to the treatment of tinea capitis with an appropriate antibiotic.

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We conducted studies on sensitivity and discrepancies in results among interpreters using five rapid influenza diagnostic test kits with different chromogenic principles, including a kit that used automatic interpretation with a dedicated device. The studies did not show any significant difference in sensitivity among the kits that used visual interpretation. The kit that used automatic interpretation via silver amplification showed significantly higher sensitivity than the kits that used visual interpretation. There were discrepancies in interpretation at the lower detection limit of each visual interpretation kit among interpreters. The automatic interpretation kit requires a dedicated device and cannot measure several specimens simultaneously, but it is considered useful in that it is more sensitive and objective than visual interpretation kits.

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Multidrug-resistant Pseudomonas aeruginosa (MDRP) is one of the most important pathogens in hospital infection control. Rapid and efficient methods of detecting MDRP are needed because early recognition of MDRP colonization/infection is important in the prevention of MDRP nosocomial spread. We evaluated the fundamental performance of CHROMagar^TM MDRP medium, a novel chromogenic agar medium, in the detection of MDRP. We cultured 31 P. aeruginosa and 30 other Gram-negative bacilli that were resistant to one or more of the following antibiotics on CHROMagar^TM MDRP medium: imipenem, ciprofloxacin and amikacin. As a result, all 8 isolates of MDRP were observed as blue-green colonies on the medium, although some of the non-MDRP P. aeruginosa isolates also grew. Among the bacteria other than P. aeruginosa, 8 of 30 strains were recovered. The observed colonies were easily distinguished from that of P. aeruginosa by the difference of color. Thus, our data suggest that CHROMagar^TM MDRP medium could be highly useful as a screening tool for detecting MDRP although antimicrobial susceptibility testing is necessary for confirmation.

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A new, particle-enhanced immunoturbidometric CK-MB assay that is applicable to most automated clinical analyzers has been developed. In this study, we compared the performance of the new method with an immunoinhibition-based enzymatic assay and a chemiluminescence-based immunoassay. The test results of the new method showed good run-to-run precision, good linearity, and good correlation with the chemiluminescence immunoassay. When CK anomalies were present in the serum of patients with malignant tumors, CK-MB activity was abnormally high and its correlation with the enzymatic assay was affected. When the patient samples with anomalies were excluded, the new method and CK-MB immunoinhibition method using anti-MtCK antibodies highly correlated. These results indicate that the new method, applicable to automated clinical analyzers, is useful for rapid measurement of CK-MB serum concentration.

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Two hundred and sixteen strains of β-hemolytic Streptococcus were detected at five institutions in fiscal years 2007, 2009, and 2011. These strains were divided into three bacterial groups: Streptococcus pyogenes (Group A Streptococcus, GAS), Streptococcus agalactiae (Group B Streptococcus : GBS), and β-hemolytic Streptococcus strains other than GAS or GBS (non-AB). Changes in detection rate and antibiotic sensitivity of these strains were surveyed. No marked change was noted in detection rate, but that of the bacterial group assumed to include Streptococcus equisimilis increased. Regarding antibiotic sensitivity, all bacterial groups were sensitive to penicillin antibiotics. Regarding erythromycin (EM) sensitivity, two of seven GAS strains (28.6%) isolated in 2011 were resistant, showing that resistant strains were present although the number of isolates was small. The EM resistance rate did not markedly change in GBS, but tended to increase in non-AB. Levofloxacin (LVFX) resistance increased yearly in GBS and reached 61.1% in 2011. Resistance was also noted in one non-AB strain in 2011. It is necessary to pay attention to the acquisition of antibiotic resistance in the future.

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The committee for laboratory data standardization of Tokyo Metropolitan Association of Medical Technologists conducts an annual quality control survey using human fresh materials to evaluate testing data from participating institutions without distinct machines and reagents. However, in serum albumin testing, there is a significant difference between values of the bromocresol green (BCG) method and those of the modified bromocresol purple (BCP) method; this testing item was not evaluated for the past two years. In this report, we analyzed the serum albumin data from the surveys conducted in 2010 and 2011. As a result, there was a large difference between the data obtained by the two methods; the difference was smaller in 2011 than in 2010. The ratio of α-globulin in survey samples was smaller in 2011 than in 2010, and similar to the ratio in each manufacturer’s standard product. Moreover, there was an obvious difference among values obtained by the modified BCP method. It is our mission to elucidate the cause of the data difference among reagents based on the modified BCP method and to further standardize serum albumin measurement.

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Autologous blood transfusion is currently considered to be the safest among blood transfusion methods. However, it still has adverse events and risks during collection, storage and transfusion of blood. To minimize the adverse events and risks, a centralized team for preoperative autologous blood collection was established in January 2010 with great effort of medical staff including doctors, nurses and technologists. In addition, a standard operating procedure of preoperative autologous blood collection in our hospital was established. As a result, potential adverse events and risks were reduced. Furthermore, we have been able to have more time to have discussion for the reduction of risks in each patient requiring autologous blood transfusion.

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