We evaluated the performance of liquid chromatography coupled with atmospheric-pressure chemical ionization (LC-APCI-MS) and electrospray ionization mass spectrometry (LC-ESI-MS) for the simultaneous extraction of five organophosphorus and two carbamate insecticides from human serum. Nonpolar and polar pesticides were simultaneously extracted from serum by solid-phase extraction using Oasis HLB columns connected with the activated carbon Jr. The chromatographic separation was performed on an Xterra MS C18 column (150 × 2.1 mm, 3.5 μm) using 0.1% acetic acid/0.05% acetic acid in methanol as the mobile phase at a flow rate of 0.2 mL/min. The assay linearity showed good results with correlation coefficients of >0.996. The lower limit of quantification was between 0.01 and 0.5 μg/mL. Interday assay reproducibility (coefficient of variation) was between 2.5 and 18.4%, and the recoveries were between 82.0 and 108.3%. The applicability of this method was also demonstrated for the analysis of serum samples from clinical poisoning cases.

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Myasthenia gravis (MG) is an autoimmune disease related to abnormalities of the neuromuscular junction, and occurs because of the presence of autologous antibodies for the acetylcholine receptor (AChR) or muscle-specific receptor tyrosine kinase (MuSK). Repetitive nerve stimulation (RNS), an assay for AChR, and the tensilon test are commonly performed to diagnose MG. We report the case of a girl aged 14 years and 8 months with generalized MG, and studied RNS during the clinical course. The patient experienced repeated remission and relapse. Waning on RNS disappeared during remission, whereas waning was observed at relapse. The results of RNS changed in accordance with her clinical course. RNS was a more sensitive test than the determination of the serum AChR value to detect her relapse.

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Agranulocytosis is defined as an absolute neutrophil count of less than 500/μL. Many drugs can cause agranulocytosis. We report a case of antibiotics-induced agranulocytosis in which information from our laboratory resulted in early treatment.The patient was a 67-year-old woman who had been diagnosed as having primary sclerosing cholangitis associated with bacterial infection. She was treated with several antibiotics. After two weeks, her leukocyte count decreased to 1.7×10³/μL and the neutrophil percentage was 0%. We immediately reported to the doctors that the patient might have agranulocytosis. Early treatment was started and further evaluation was performed.Drug-induced agranulocytosis is considered to be caused by immunological or toxic mechanisms. Previous reports have shown that maturation arrest may be observed in patients with agranulocytosis mediated by toxic mechanisms. In this patient, bone marrow examination showed maturation arrest at the myelocyte stage, and this finding suggested that drug-induced agranulocytosis mediated by toxic mechanisms occurred.Because patients with agranulocytosis are likely to show complications caused by severe infections, an early detection of agranulocytosis is very important. Inadequate follow-up may occur in patients under treatment with drugs that rarely cause agranulocytosis. Quick reports to doctors may be required for a rapid diagnosis of antibiotics-induced agranulocytosis.

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The patient was a 29-year-old male who visited our hospital complaining of pain from the abdomen to the back through the left side. Chest-abdominal CT showed a large number of nodules and enlarged lymph nodes with right pleural effusion and ascites. PET-CT showed FDG uptake of advanced pleural and peritoneal ascites. Levels of serum CEA, CA125 and sIL-2R were increased. From the above results, it was necessary to distinguish among peritonitis carcinomatosa, malignant lymphoma and malignant mesothelioma of tuberculous peritonitis. Since it was difficult to establish a definitive diagnosis, laparoscopic biopsy of the peritoneum was performed. A large number of white nodules were diffused in the peritoneum, the surface of the liver and the greater omentum. The parietal side of the peritoneum closely adhered to the greater omentum. On the basis of rapid perioperative histopathological diagnosis, the patient was diagnosed as having tuberculous peritonitis. Tuberculous peritonitis is rare. Thus, the diagnosis is difficult based on clinical findings and examinations. An exploratory laparoscopy and rapid perioperative histopathological examination would be effective for accurate diagnosis.

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Acute renal failure resulting from rhabdomyolysis is caused by the release of myoglobin into the circulation, leading to tubular epithelial cell injury or tubular lumen obstruction by myoglobin casts. Morphologically, myoglobin casts are similar to granular and waxy casts, and are difficult to distinguish by urine sediment analysis. In the diuretic phase of myoglobinuric acute renal failure, myoglobin casts may be contained in granular and waxy casts. We describe a case of myoglobin casts contained in waxy casts in the diuretic phase of acute renal failure resulting from rhabdomyolysis.

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Acquired cystic disease of the kidney (ACDK) has a strong relationship with dialysis. The incidence of renal cell carcinoma is high in ACDK. However, the cytologic features of ACDK-associated renal cell carcinoma (RCC) in voided urine have rarely been reported. A 37-year-old male with a long history of IgA nephropathy presented with a both-kidney mass, detected by magnetic resonance imaging. Urine cytology smears showed moderately cellular characteristics and were composed of three-dimensional clusters and single cells. These atypical cells showed abundant foamy cytoplasm, and the nuclei were round and showed fine chromatin, with single or multiple small nucleoli. In addition to atypical cells, smears showed many isomorphic erythrocytes in the background. In immunocytochemistry, these atypical cells showed positivity for CD15. The diagnosis of ACDK-associated RCC was established from the subsequent histologic examination of both kidneys. When urine cytology from long-term dialysis patients reveals a cluster of atypical cells with abundant foamy cytoplasm, finely granular chromatin, and nucleoli, one should suspect an ACDK-associated RCC. The correlation of clinical presentation and imaging analysis with cytologic findings is important, and can be further supplemented by immunocytochemistry to differentiate it from other tumor cells.

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Hemoglobinopathy is defined as mutations that change the amino acid sequence of one of the globin chains. More than 800 hemoglobin variants have been identified in the world. The frequency of mutant variants is estimated to be about one per 3,000. Recently, hemoglobinopathies have been found among individuals with a high or low hemoglobin A_1c level or with an abnormal chromatogram pattern. We encountered hemoglobinopathy of four patients in two unrelated families. The disease is hereditary and it is necessary to recognize its appearance in areas with little migration of inhabitants. As hemoglobin A_1c may be measured even if it is an unusual chromatogram, we require attention to the chromatogram pattern. Although it is necessary to fully consider gene analysis, in order to avoid unnecessary inspection and medical treatment, we believe that it is important for patients to understand their own morbidity. We must offer suitable information and recommend an appropriate examination for the clinical aspect.

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C-reactive protein (CRP) is often used as a marker of bacterial infection, tissue injury, and inflammatory diseases. Recent studies have shown that inflammation plays an important role in the development of atherosclerosis, which causes cardiovascular disease (CVD); therefore, CRP has received considerable attention as a risk marker of CVD. In this study, we evaluated the basic performance of three CRP assay reagents, which reportedly have comparable performance to the high-sensitivity CRP (hsCRP) reagent. Each reagent had a wide dynamic range (approximately 35 mg/dL), which was sufficient for its use as a conventional inflammation marker with satisfactory precision, accuracy, and reproducibility. The three reagents that we evaluated in this study could be clinically used in routine assay kits at a performance level comparable to that of the hsCRP assay reagent.

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We performed basic evaluation studies of the reagent ‘auto RF·BML’ for rheumatoid factor (RF) measurement by the latex turbidimetric immunoassay. This reagent showed that CVs of within-run precision were less than 2.0%, CVs of between-day precision were less than 3.0%, the detection limit was 2.5IU/mL, and the dilution linearity was also good. No effects from interfering substances were detected. In addition, its correlation with the comparison product ‘Iatro RF II’, the measurement principle of which is similar to that of this reagent, was good (y = 1.055x + 1.4, r = 0.995, n = 315). Moreover, its correlation with the comparison product ‘Auto TIA RF [Nissui]’, the measurement principle of which is the turbidimetric immunoassay, was also good (y = 1.066x + 4.0, r = 0.983, n = 342). ‘Auto RF·BML’ showed acceptable reagent performance in routine tests.

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We examined the usefulness of the dual-color in situ hybridization (DISH) method for HER2 gene amplification in breast cancer. We studied 27 cases of invasive breast cancer that scored 2+ by the HER2 immunohistochemistry (IHC) method. We investigated the occurrence of HER2 gene amplification by fluorescence in situ hybridization (FISH) and DISH methods, and compared the results obtained. Four investigators carried out the determinations by the HER2 DISH method and the results were compared. Comparison of the results obtained by the four investigators using the DISH method showed that 10 cases (37%) revealed a small dispersion in the HER2 gene/CEP17 signal ratios among the investigators, but the difference in the determined results was not confirmed in any of the cases. The FISH and DISH methods for HER2 gene diagnosis showed uniformity in 21 cases (78%). Inconsistencies were prevalent in 6 cases (22%). The HER2 gene/CEP17 signal ratio obtained by the DISH method significantly correlated with the result of the FISH method.

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The detection of inducible clindamycin (CLDM) resistance in Staphylococcus by the microdilution method was added to M100-S18 of the Clinical and Laboratory Standard Institute (CLSI). In response, for this revision, BD Phoenix^TM Gram-Positive PMIC/ID74, a panel for gram-positive bacteria with an additional new iMLSB well, was developed for BD Phoenix^TM, an automatic identification-sensitivity test system. This has allowed the detection of inducible CLDM resistance by the iMLSB test without additional tests in parallel with identification-sensitivity tests. Therefore, the basic performance of the iMLSB test was evaluated in 120 isolates of Staphylococcus. The agreement between the iMLSB test and the control test (D-test) for resistance was 99.2%, yielding superior results. The use of this panel was considered to be advantageous, allowing more rapid reporting on CLDM-inducible resistance than the conventional method.

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We evaluated the ‘L-type Wako CK-MB mass’ reagent in terms of fundamental performance. This reagent can be used to measure CK-MB protein values in a short time on an automated analyzer. In this study, the reagent exhibited satisfactory performance in terms of sensitivity, with-run precision, between-day precision, linearity, and effects of interfering substances. This reagent showed good correlation with the chemiluminescent enzyme immunoassay and the immunoinhibition method. This reagent did not have an effect on macro-CK type 1 and mitochondrial CK, but have little effect on CK-BB. Although extremely rare, we found that heterophile antibodies affected the results obtained using this reagent. On the basis of the present results, we conclude that the reagent performs sufficiently well in routine laboratory tests.

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Teicoplanin exhibits antibacterial activity against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), and monitoring its serum concentration under the “Guideline for therapeutic drug monitoring of antibiotics” is encouraged. In this study, the fundamental performance of the Teicoplanin assay “Nanopia^® TDM Teicoplanin” was evaluated. The interferences of C-reactive protein (CRP), total cholesterol (TC), and rheumatoid factor (RF) were also evaluated. The C.V. (%) values of within-run and between-run precisions were determined to be 1.60–6.59% and 0.46–11.95%, respectively. Deviations from the target values were observed toward the low end of approximately 10 μg/mL. The linearity was good and no prozone phenomenon was observed up to 100 μg/mL. There was no significant interference by CRP, TC, and RF observed in the analytical recovery study. The measured values of the patient samples became lower when saline was used for sample dilution, suggesting that the ideal diluents would be either special dilution buffer or Teicoplanin-free serum. Comparison with fluorescence polarization immunoassay (FPIA) showed good correlation with a regression equation of y = 0.94x + 1.87 and a correlation coefficient of r = 0.986.

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Recently, carbapenem-resistant Gram-negative bacteria belonging to the family Enterobacteriaceae (CRE) have become a general clinical concern, but it is very difficult to elucidate the molecular mechanisms underlying their carbapenem resistant phenotype. In this study, three Escherichia coli strains (NUBL-5310, NUBL-5317, and NUBL-9600), two Klebsiella pneumoniae strains (NUBL-5307, and NUBL-5309), one Enterobacter aerogenes strain (NUBL-5311), and one Enterobacter cloacae strain (NUBL-7700) demonstrating carbapenem nonsusceptibility or cephem resistant phenotypes were subjected to an antimicrobial susceptibility test, β-lactamase typing using β-lactamase inhibitors, and polymerase chain reaction (PCR). Conjugal transfer of plasmid, transformation, gene cloning, and nucleotide sequence analyses were additionally performed in some isolates. In E. cloacae NUBL-7700, genes for bla_IMP and bla_{CTX-M-1 group} were detected by PCR. In the remaining six strains exhibiting reduced susceptibility or resistance profiles to carbapenems, no new carbapenemase was detected, but genes for CMY-, DHA-, or CTX-M-type β-lactamases were detected in six isolates. Increased production of chromosomal AmpC was speculated in E. aerogenes NUBL-5311 because aminophenyl boronic acid reduced the resistance to cephems and no genes for plasmid-mediated β-lactamases were detected by PCR. Augmented production of CMY-, DHA-, or CTX-M-type β-lactamases and the decrease or deletion of certain outer membrane proteins probably contribute to the carbapenem-nonsusceptible phenotypes among the isolates tested.

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Aspergillus is a fungal genus causing deep mycosis. We isolated 344 strains (320 cases) of Aspergillus species from clinical specimens obtained from the Nagaoka Red Cross Hospital between 2000 and 2011. We evaluated the species, isolation frequency, underlying diseases, and relevance of microscopic and serologic tests. We isolated 126 strains of Aspergillus niger (36.6%), 113 strains of A. fumigatus (32.8%), 53 strains of A. terreus (15.4%), 22 strains of A. flavus (6.4%), and 30 strains of other Aspergillus species (8.7%). Interestingly, we found that A. fumigatus accounted for 46.0% of respiratory specimens, whereas A. niger accounted for 48.3% of otolaryngology specimens. Recently, the rates of isolation of A. fumigatus and A. niger from respiratory samples have been demonstrated to have increased. Many patients in whom Aspergillus species were isolated from the respiratory tract had respiratory disease as the underlying disease. The clinical significance of the isolation of Aspergillus species from clinical samples should be determined on the basis of the underlying disease and clinical findings, including those from chest images and serologic tests. The emergence of resistant Aspergillus strains and the differences in drug sensitivity between species have also been reported, indicating that continuous attention to this issue will be necessary in the future.

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A virtual slide is used in the imaging of a part or the whole of a specimen. We freely change the observation view and magnification, and we can observe a specimen using a computer as in the case of using a microscope. We report on the clinical use of the virtual slide during the urinary sediments workshop. We made a virtual slide using VS120. Participants observed a glass specimen under a microscope, and the presenter expounded on the use of the virtual slide. Many participants had few years of experience and had not used an autoanalyser. Therefore, we considered a virtual slide to be useful as a training tool for screening. We used the virtual slide during the workshop for the first time and it was popular among the participants. We expect that a virtual slide can be applied to various uses besides a workshop, for example, for precision control survey and the permanent preservation of rare specimens.

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The QuantiFERON (QFT) test is one of the interferon-γ release assays (IGRA) used as a specific diagnostic method for tuberculosis (Tb) infection. We investigated the status of QFT for the past 5 years and analyzed the data in our hospital. The QFT-positive rate was high in elderly patients. Undeterminable results were often obtained in immunosuppressed patients or those undergoing hemodialysis. We compared the data of QFT with those of acid-fast bacillus (AFB) tests (smear, culture and PCR examinations) and found that none of the QFT-negative cases revealed the detection of Tb. On the other hand, of the 157 QFT-positive cases, 18 cases showed Tb infection and 125 cases showed negativity in AFB tests. Next, we compared each QFT value with the results of AFB tests and found that the mean values of QFT seemed to increase in the order of AFB tests, namely, negative group, non-Tb AFB-positive group, and Tb-positive group, while there was no statistical difference between the groups. Considering that Tb was not detected in any of the QFT-negative cases in the following AFB tests, we found that QFT is worth conducting as the first screening test for the diagnosis of Tb. However, in QFT-positive cases, the patients require comprehensive medical examinations to determine the presence of Tb infection.

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We determined the attitudes of high school students towards sexually transmitted diseases using a questionnaire survey. Replies were obtained from 1,548 high school students who consented to the details of the research. It was hypothesized, on the basis of the degree of comprehension of sexually transmitted diseases, that there is a correlation between consciousness of sexually transmitted diseases and first sexual experience. Concerning new knowledge of sexually transmitted diseases, many answered that infection is not noticed in many cases. Given the present condition, the present young age group is in a dangerous situation. The degree of comprehension to the importance of condom use was high regardless of a grade or gender. There were many opinions concerning pregnancy or childbirth from female students, regardless of grade. There were also many comments concerning prevention methods from male students.

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Anaerobic bacteremia is uncommon since bacteremia is generally caused by aerobic or facultative bacteria. Anaerobic bacteria are found on the gastrointestinal mucosa and can cause polymicrobial bloodstream infection. However, the association between anaerobic bacteria and polymicrobial bloodstream infection is still unclear. In this study, we investigated the proportion of isolated anaerobic bacteria and the combination of anaerobic bacteria in polymicrobial bloodstream infection. During the 2-year study period, 738 episodes yielded bacteremia. Polymicrobial bloodstream infections were 83 episodes (83/738, 11.2%). Twenty-one out of the 83 episodes (21/83, 25.3%) include at least 1 anaerobic bacterium. A total of 36 anaerobic bacteria were cultured during 21 episodes of polymicrobial bloodstream infection. The most abundant anaerobic species were the Bacteroides fragilis group (11/36, 30.6%) and Clostridium species (7/36, 19.4%). The most common combinations of anaerobic bacteria in polymicrobial bloodstream infection were those comprising only anaerobic bacteria (9/21, 42.9%). Other combinations with anaerobic bacteria included gram-negative bacilli (4/21, 19.0%) and gram-positive cocci (5/21, 23.8%). The results of this study suggest that anaerobic bacteria are commonly found in polymicrobial bloodstream infection. Inappropriate antimicrobial therapy for patients with anaerobic bacteremia leads to poor prognosis. Polymicrobial Gram-staining findings or positive anaerobic blood culture underscores the need for the identification of anaerobic bacteria and for clinicians to choose an antibiotic treatment keeping anaerobes in mind.

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In recent years, the number of patients with amebic colitis, who are homosexual males or inmates of mental retardation facilities in many cases, have been increasing gradually. Although ameba detection in feces or serum antibody tests are currently used for diagnosis, there are some problems, namely, feces provide poor detection sensitivity and serum antibody tests may not always reflect the current infection status. We, therefore, performed direct microscopy on biopsy tissues by colonoscopy to investigate its usefulness for the diagnosis of amebic colitis. From June 2012 to April 2013, colonoscopies were carried out and biopsy tissues were harvested from patients who have lesions with ulcers or white mosses in the colon. Three specimens with amoeboid trophozoites were detected in six colonoscopies during the study period. While pathological examinations and serum antibody tests were also performed simultaneously, there were some discrepancies among their outcomes. Three patients judged positive were finally diagnosed as having amebic colitis and the treatments were started immediately. The results of direct microscopy on biopsy tissues obtained by colonoscopy were all available within 30 min. In conclusion, direct microscopy on biopsy tissues is effective for diagnosing amebic colitis in a general laboratory.

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