Aim: In Azan staining, reagents considered as powerful drugs or carcinogenic substances are used. To avoid the use of harmful reagents, it is necessary to improve the staining quality of azocarmine G solution. We developed new methods of preparing azocarmine G solution to obtain improved staining results. Methods: We prepared azocarmine G solutions by using two methods for dissolving sparingly soluble materials. For comparison, a freshly prepared azocarmine G solution was made. The staining qualities of these solutions were compared with that of a commercially available azocarmine G solution. Result: The staining qualities of the azocarmine G solutions produced by the method of dissolving sparingly soluble materials were excellent. In particular, the azocarmine G solution produced by microwave heat decomposition showed a better staining result without requiring differentiation or heating than the commercially available staining solution that requires differentiation. Conclusion: Using azocarmine G solution produced by microwave heat decomposition, we can avoid the use of harmful reagents during Azan staining. Furthermore, we can largely simplify the staining procedure using this solution.

WBC-DIFF scattergrams obtained from newborn blood samples using an XE2100 multiparameter automatic hematology analyzer show low side fluorescence (SFL) intensities. We found six adult blood samples that exhibited significantly low SFL intensities. The affected and newborn blood cells were divided into red blood cells (RBCs), white blood cells (WBCs), and mixed with WBCs or RBCs separated from the blood of normal adults. Extremely low SFL intensities determined using XE2100 were observed for paired samples of newborn RBCs or affected RBCs mixed with normal WBCs. Our previous studies indicated that hemoglobinopathy showed abnormal WBC scattergrams obtained using an NE7000 multiparameter automatic hematology analyzer. Therefore, we speculated that extremely low SFL intensities obtained using XE2100 may have been due to some variations in RBCs or hemoglobinopathy; thus, we performed genetic sequencing analyses of globin genes from four of the six affected patients. Three patients showed HbJ-Calabria-type mutation and the other had Hb Yamagata-type mutation. It is of interest that hemoglobinopathy was found on the basis of DIFF scattergrams obtained using XE2100. However, the mechanism underlying the extremely low SFL intensities in hemoglobinopathy remains unclear.

【Objectives】The advantages of using liquid-based cytology (LBC) in the diagnosis of cervical squamous lesions are well established. However, the possible advantages of conducting endometrial LBC have not been fully explored. The aim of this study was to clarify the characteristics of endometrial LBC using microscopic cell images and by image analysis. 【Materials and Methods】We conducted the LBC of G1 and G2 endometrial carcinomas. We determined the numbers of component cells and overlapping layers in the clusters using microscopic cell images and by image analysis. Then, the nucleolus/nucleus area (N/N) ratios obtained by digital analysis in conventional measurement methods and LBC were compared. 【Results】The numbers of component cells and overlapping layers were lower in the G2 cluster than in the G1 cluster. Moreover, the number of overlapping layers obtained by image analysis was smaller than that obtained by examination of microscopic cell images. The nuclear and nucleolar areas in both the G1 and G2 endometrial cancer cells determined by LBC were significantly smaller than those determined by the conventional method. Conversely, the N/N ratio determined by LBC was larger than that determined by the conventional method. 【Conclusion】It is advantageous to employ LBC as a standard method of endometrial cytology. The establishment of new diagnostic criteria for endometrial LBC could lead to an increased accuracy of endometrial cytology in the future.

Domestically, the number of diabetic patients is increasing. As a means of daily glycemic control by patients, devices for the self-monitoring of blood glucose (SMBG) are increasingly gaining importance. Many SMBG devices are being sold by manufacturers, and their measurement accuracies are achieved in accordance with ISO15197, which was revised in 2013. However, it has been reported that the measurement accuracies differ between models. In this study, medical staff members perform tests using selected models in order to contribute to their improvement so that patients can use the SMBG devices with peace of mind. They also conducted precision tests using 7 models of the SMBG devices and evaluated their basic performance on the basis of the new criteria [ISO15197 (2013)].

Recently, standardization and transparency have been demanded in various industry fields, and their importance is increasing in the medical field. ISO 15189 is an international standard of clinical laboratories, and the Japan Accreditation Board (JAB) authorizes it. However, for the introduction and maintenance of ISO 15189, complicated office work such as preparing large amounts of documents is required, and balancing ISO 15189 with routine work is important. Therefore, we developed an original management support system adapted to ISO 15189 to simplify the complicated office work. Using the management support system enabled the digitization of documents and the unification of information, and office work became expedited. The management support system that we developed contributed to the efficiency of office work and was useful in the introduction and maintenance of ISO 15189.

Protein induced by vitamin K absence or antagonist-II (PIVKA-II), a tumor marker of hepatocellular carcinoma, is the abnormal variant of pro-thrombin produced in the absence of vitamin K. In this study, we evaluated the basic performance of a new assay kit, ARCHITECT PIVKA-II, in measuring PIVKA-II. The coefficients of variation (C.V.) of within-run and between-day precisions with quality control samples for ARCHITECT PIVKA-II were 2.2 to 3.0% and 2.8 to 4.9%, respectively. Using pooled sera of patients, we determined the C.V. to be 2.0 to 2.7% and 3.2 to 4.9%, respectively. It showed good linearity from 0 to 30,000 mAU/mL, and the limit of quantitation was 3.45 mAU/mL. No interference by endogenous substances was observed. The correlations between Picolumi or Lumipulse Presto and ARCHITECT PIVKA-II assay were good. The differences in the values of the samples prepared using plain serum tubes, serum separator tubes, and rapid serum tubes, which include thrombin, were not significant. These results indicate that ARCHITECT PIVKA-II has high basic performance and greatly contributes to routine tests in laboratories.

Parathyroid hormones (PTHs) are analyzed by intact-PTH or whole-PTH assay. The intact-PTH assay detects PTH (1–84) and PTH (7–84), whereas the whole-PTH assay detects only PTH (1–84). The whole-PTH assay could be performed using only the immunoradiometric assay (IRMA) method, but now, it can also be performed using CLEIA and ECLIA with an automatic analyzer. Therefore, we performed a basic study of CLEIA and ECLIA and compared the two methods. Both methods have good repeatability and intermediate precision. However, there was a high tendency of ECLIA to show dilutional linearity in the low concentration range. No effect of interfering substances was observed up to a final concentration of each substances. The preservation stabilities for both methods are poorer in serum than in EDTA-2Na plasma, and PTH value was most unstable at room temperature. The IRMA method has a lower PTH value than CLEIA or ECLIA. Changes in the trend of the concentration band are seen in the comparison between CLEIA and ECLIA. A good correlation was obtained in the comparison between serum and EDTA-2Na plasma in CLEIA and ECLIA. Higher concentrations of PTHs determined using MPC-treated containers than using untreated containers were confirmed. Since the adsorption reaction to the container is known to be suppressed by applying the MPC process, this phenomenon was considered to be due to the adsorption of PTH to the storage container. The IRMA method, which is a conventional method with relatively high stability, was shown in the present study to be useful in combination with CLEIA.

This study was conducted between July 2014 and March 2016 to determine the prevalence of extended spectrum β-lactamase (ESBL)-producing Enterobacteriaceae in the fecal samples from 111 outpatients, who provided their informed written consent, in the Department of Infectious Diseases of Tokyo Metropolitan Bokutoh Hospital. ESBL-producing Enterobacteriaceae isolates were detected in 32 fecal samples: Escherichia coli in 31 (27.9%) and Klebsiella pneumoniae in one (0.9%). The prevalence of ESBL-producing organisms was significantly higher (p < 0.05) in individuals with a history of recent travel (48.8%) than in those without it (16.2%). Although patients had a history of travel to a variety of areas including Asian, African, Middle-Eastern and South American countries, travels to India were the most common. In the case of hospitalization of patients with a history of recent overseas travel, hospitals should strengthen surveillance to prevent an outbreak of nosocomial infection because of the high possibility that these patients might be carriers of ESBL-producing Enterobacteriaceae.

We report the case of a 74-year-old man who presented with a tuberculoid granuloma in the scrotal skin after bacillus Calmette–Guérin (BCG) therapy. The patient was treated for bladder cancer with transurethral resection of the bladder tumor, MVAC (methotrexate, vinblastine, adriamycin and cisplatin) chemotherapy, and BCG therapy from 2008 to 2015. After the second BCG treatment, the patient found a tubercle in the right scrotal skin and visited the Department of Dermatology at our hospital. A Mycobacterium sp. was isolated from the biopsy sample. PCR and immunochromatography revealed that the isolated Mycobacterium sp. belonged to the Mycobacterium tuberculosis complex. However, it is necessary to distinguish M. tuberculosis from M. bovis BCG because the treatment methods and infection control measures for the two organisms differ. We performed another PCR, which showed that the M. tuberculosis complex strain was M. bovis BCG. Most strains of the M. tuberculosis complex, which usually infect humans, are identified as M. tuberculosis; and infection by M. bovis BCG is quite rare. However, we should consider M. bovis BCG in cases of urological infection in patients with bladder cancer.

Roseomonas is a pink-pigmented, nonfermentative, Gram-negative bacillus having clinical importance as an opportunistic pathogen that can lead to infection, especially in immunosuppressed individuals. Here, we report a case of bacteremia caused by R. gilardii subsp. gilardii from blood culture. An 82-year-old man was diagnosed as having pneumonia and was hospitalized. When the patient developed fever after ten days of hospitalization, blood was collected for two sets of bacterial culture. Gram-negative bacillus cells were observed in one of the sets, and pink mucoid colonies grew on sheep blood agar, chocolate agar, and modified Drigalski agar. The strain was identified using automated WalkAway40SI and IDtestNF-18. However, it could not be parsed. By sequencing analysis of the 16S rRNA gene, the bacillus was identified as R. gilardii subsp. gilardii. Sulfamethoxazole-trimethoprim was administered on the basis of the antimicrobial susceptibility results and the patient’s condition improved. In many laboratories, it is difficult to identify the species of the genus Roseomonas by routine microbial tests. Therefore, their clinical significance in immunocompromised hosts is not known. However, it is important to integrate the antimicrobial susceptibility and infection treatment data of Roseomonas infection.

We report a case of invasive pneumococcal disease (IPD) that was presumably diagnosed by Gram staining of blood in culture bottles still under incubation. A man in his sixties with chief complains of generalized fatigue and chill was brought to our Emergency Department. On arrival at the hospital, spreading purpura was observed in his whole body, and his general condition deteriorated rapidly after admission. To identify the causal pathogens as soon as possible, we performed Gram staining of a blood culture before an automated system showed positive results. The Gram staining showed Gram-positive diplococci, and the rapid urinary antigen test was positive for Streptococcus pneumoniae. We finally diagnosed him as having IPD earlier than the usual process but he died without any improvement of his clinical condition. This case indicates that Gram staining of cultured blood can be used to detect pathogenic bacteria earlier than the usual blood culture system. Although there are some issues that need to be resolved before implementing this method, it is clearly worth considering for further investigation.

An 80-year-old woman presented with leukocytosis. Laboratory blood data showed a WBC count of 11,100/μL (LY 71.5%), a Hb level of 12.6 g/dL, and a PLT count of 29.1 × 10⁴ /μL. Flow cytometric analysis results showed CD2+, CD3+, CD4+, CD5+, CD7+, and CD8+ cells. She was diagnosed as having T-PLL. Shortly thereafter, her condition worsened and treatment was started. Fludarabine as the first treatment or THP-COP as the second treatment was not effective. Then, Alemtuzumab (ALZ) treatment was started, which is a new drug against refractory CLL. Three days after starting the treatment, her WBC count became normal and her condition was improved. The duration of administration was 12 weeks, which was within the guidelines. Two months after the final dose, her condition became stable and she is now under treatment. This time, we obtained a result that makes us expect a recovery from T-PLL, which is considered a refractory disease. Now, it is very important in terms of quality assurance that we understand the causes of changes in laboratory data and correct information about new treatments and drugs.