In recent years, the treatment of invasive breast cancer (IBC) has been based on intrinsic subtype classification by gene expression profiling (GEP). Because GEP is very expensive for every single case of IBC, immunoprofiling has been widely used as an alternative method of classification. Although the opportunity of cytological examination has declined despite its advantages of being non-invasive, inexpensive, and time-saving, we evaluated the usefulness of the cytological approach for IBC diagnosis compared with the alternative intrinsic subtype classification based on immunohistochemistry. This study was performed on 97 cases of histopathologically confirmed IBC whose stamp preparations were available at the time of needle biopsy. We used six nuclear parameters with a value of 1 or 2, namely, the size of the nucleus, nucleus-to-cytoplasm ratio, nuclear irregularity, chromatin status, chromatin intensity, and degree of anisokaryosis. We also employed an architectural parameter with a value of 1, 2 or 3. The sum of each parameter is plotted on the basis of histological types classified by an alternative method. It was found that cases with a total score of less than 8 are highly likely to be in the luminal A-like group, whereas about 74% of IBC cases with a score of 13 or more are either in the luminal B-like (HER2-positive), HER2-enriched, or triple negative group. Our results suggest that the cytological method utilizing our seven parameters may be useful for the evaluation of the treatment and prognosis of IBC patients.
With the widespread use of molecular biological techniques, Mycobacterium species were reclassified into five genera, and the number of bacterial species increased to more than 188. Thus, it is difficult to identify nontuberculous mycobacteria by conventional genetic and phenotype tests. Therefore, we attempted to identify Mycobacterium species by phylogenetic tree analysis and BLAST (Basic Local Alignment Search Tool) analysis of secA1, which is one of the housekeeping genes. As a result, it was possible to identify Mycobacterium abscessus, Mycobacterium avium, Mycobacterium fortuitum, Mycobacterium lentiflavum, Mycobacterium mageritense, Mycobacterium marseillense, and Mycobacterium ulcerans, which were difficult to identify by the DDH method or 16S rRNA gene analysis. Although there are still a few problems in the identification of Mycobacterium intracellulare, the increase in the numbers of reference and standard strains registered in GenBank has expanded its usefulness as a tool for identifying nontuberculous mycobacteria.
Recently, the Japanese version of the Montreal Cognitive Assessment (MoCA-J) has been disseminated as a screening method for patients with mild cognitive impairment (MCI). We investigated the relationship between MoCA-J and MRI findings in cognitively normal patients. In addition, we compared the subscores of MoCA-J among various patients with cognitive impairment. We enrolled 75 patients (age, 74.6 ± 9.1 years; 30 females) who visited the outpatient memory care ward from August 2018 to March 2019. The median MoCA-J score was 21 (minimum 8, maximum 30). Among cognitively normal patients (n = 39), periventricular hyperintensity (cerebral white matter lesions) on MR images was independently associated with the MoCA-J score by multivariate analysis. For subscores of MoCA-J, attention and executive function subscores were markedly lower in vascular dementia patients. However, subscores of memory were lower for all the subjects because the memory task was difficult. The MoCA-J score consisted of orientation, memory, visuospatial function, language, attention, and executive function. The proportions of attention and executive function were large, which can be used for the diagnosis of frontal lobe dysfunction. In addition, the memory task was difficult, which made it easier to detect slight memory loss. Because of these characteristic features, MoCA-J is useful for evaluating various cognitive impairments.
The fraction of exhaled nitric oxide (FENO) is determined by various factors, but only a few studies have examined its environmental factors, which include seasonal variations in Japan. Therefore, to understand the pathology of bronchial asthma (hereinafter referred to as asthma), we investigated the characteristics of FENO as a useful biomarker of asthma. On the basis of the results of 396 FENO measurements performed from March 1, 2017 to February 28, 2018 on patients at our hospital, smoking status, sex, age, and three most common diseases were retrospectively investigated. NIOX MINO was used as the measurement device. Comparison of age and smoking status revealed that men in their 50s and 60s had higher FENO values, and smokers had higher FENO values than nonsmokers. When the seasons were categorized into four quarters, namely, from March to May (spring), June to August (summer), September to November (autumn), and December to February (winter), comparison of two groups (spring and winter) revealed a significant difference, with higher FENO values obtained during spring than during winter. Patients with asthma alone, those with asthma and allergic rhinitis (hereinafter referred to as asthma + allergy), and those with asthma and chronic obstructive pulmonary disease (hereinafter referred to as asthma + COPD, including asthma and COPD overlap) had significantly higher FENO values during spring. Examination at our hospital also confirmed that FENO values fluctuated during early spring, consistent with the findings described in international reports. In the present investigation, although it was unclear if inflammation of the respiratory tract was due to seasonal exacerbation or environmental factors, such as pollen, the results sufficiently reflected the seasonal increase in FENO values, and it appears that as a background factor, high FENO values were a major factor for the exacerbation of some cases of asthma + COPD.
Background—Pulmonary thromboembolism (PTE) and deep vein thrombosis (DVT) are generally associated with VTE. The plasma D-dimer has been used for the exclusive diagnosis of DVT, but little is known about its clinical significance in the association between the site of thrombus detected by lower limb vein sonography and the thrombotic property in VTE patients and PTE onset. Methods and Results—Sixty-six patients with VTE (mean age, 71.1 ± 14.6 years; male, 31; female, 35) were included in this study. The D-dimer level in the VTE patients with the thrombus of the central type was higher than that of the peripheral type (12.40 (7.89–25.30) μg/mL vs 4.59 (2.43–7.58) μg/mL, p < 0.01), and the thrombotic property of the acute thrombus was higher than that of the chronic thrombus (13.74 (8.29–28.05) μg/mL vs 4.74 (3.09–8.40) μg/mL, p < 0.01). Additionally, patients in the PTE group showed higher D-dimer levels than non-PTE group (16.32 (8.53–26.94) μg/mL vs 7.83 (4.10–13.57) μg/mL, p = 0.01). Receiver operating characteristic curve analysis showed that the cut-off of the D-dimer level was 8.04 μg/mL in VTE patients with PTE onset. Conclusion—It is suggested that the high plasma D-dimer level is related to the PTE onset and the high possibility of central type and acute thrombus in patients with VTE.
Serological diagnosis of novel coronavirus (SARS-CoV-2) infection (COVID-19) is expected to be used as an adjunct diagnostic method. There are two main types of anti-SARS-CoV-2 antibody reagents that detect antibodies to the spike protein S1 domain (S) and nucleocapsid protein (N) of SARS-CoV-2. In this study, we tested anti-SARS-CoV-2 antibody reagents using S and N antigens in longitudinal samples from patients diagnosed as having COVID-19. EUROIMMUN S-IgA and IgG reagents and VITROS S-total and IgG reagents were used as anti-SARS-CoV-2 antibody reagents tested using the S antigen. ARCHITECT N-reagent and cobas N-reagent were used as anti-SARS-CoV-2 antibody reagents using N antigen. The results showed that IgA antibodies to EUROIMMUN S-IgA reagent were positive from the time of hospitalization (day X). Anti-SARS-CoV-2 antibodies to VITROS S-total reagent were positive from the X + 5th day of hospitalization. IgG antibodies to ARCHITECT N-reagent were positive from the X + 8th day. In this case, the anti-SARS-CoV-2 antibody reagent tested using the S antigen showed positive results from early in the course of the disease. In particular, the anti-SARS-CoV-2 S-IgA antibody was suggested to be potentially useful as an adjunct for the early diagnosis of COVID-19.
As the novel coronavirus disease (COVID-19) pandemic evolves, anti-SARS-CoV-2 antibody (Ab) testing has emerged as an additional or alternative tool for COVID-19 diagnosis. The two antigens used in serological test are the nucleocapsid protein (N) and the spike protein S1 domain (S). In this study, we aimed to validate the performance of seven types of commercial lateral flow immunoassay (LFIA) kits and four types of antibody detection reagents using autoanalyzers as serological testing for COVID-19. Seven types of LFIA kits were purchased from five companies, Kurabo, RayBiotech, Innovita Biological Technology, LumiQuick Diagnostics, and Lepu Medical Technology. Four types of antibody detection reagents were obtained from three companies, Abbot, Roche Diagnostics, and Ortho-Clinical Diagnostics. We tested sequential sera from two patients diagnosed as having COVID-19 by reverse transcription (RT)-quantitative polymerase chain reaction (qPCR). The IgG Ab against the SARS-CoV-2 N protein was detected earlier than IgM Ab, whereas IgM and IgG Ab against the SARS-CoV-2 S protein were detectable from the early stage of infection. These results suggest that S-based Ab detection has a potential for the early detection of SARS-CoV-2 in hospitals, clinics, and test laboratories.
Background and Aim: In pathological diagnosis, Masson trichrome (MT) staining is one of the important staining methods. However, its dyeing process is complex and takes approximately 1 h. In this study, we examined a rapid method of MT staining. Methods: The dyeing process simplification, preparation of dyes, and time reduction effect of microwave (MW) were examined, and the dyeing result was compared with that of the conventional method. Results: The time of the rapid method was 1 min each for iron hematoxylin and the orange G/acid fuchsin mixture, and MW irradiation was 10 s each for phosphotungstic acid (PTA) and aniline blue (after 3 and 7 min for kidney and liver, respectively). Moreover, good staining results similar to the conventional method were obtained by the rapid method. Furthermore, to simplify the dyeing step, the steps of the first mordant, 1% hydrochloric acid alcohol, the second mordant, and 1% acetic acid aqueous solution were omitted. As a result, no decrease in staining property was observed. Conclusion: In MT staining, the simplification of the dyeing process and the reduction in time were realized. This is the shortest time among the presently reported staining methods. Time reduction was suggested to be made possible by the elimination of intermolecular competition in cytoplasmic staining and the enhancement of the mordanting effect of PTA by MW. Therefore, the rapid method can be used for clinical diagnosis by obtaining staining results equivalent to those of the conventional method.
The lactate dehydrogenase (LD) activity test was converted from the JSCC method to the IFCC method on 1 April 2020. We conducted a basic evaluation of LD activity using “L-type Wako LD·IF” manufactured by Fujifilm-Wako. Parallel-run precision and reproducibility for all control materials were good with a C.V. below 1.2%. The detection limit determined using 10 dilution series of high-activity LD samples was 2,470 U/L. There was no interference by bilirubin F, bilirubin C or chyle, but hemoglobin caused a false positive result. The correlation between currently used reagents was good with similar results, that is, correlation factor r = 0.994 and regression equation y = 0.923x + 9.8. Isozyme analyses of five samples using “Quick Gel LD” manufactured by Helena showed a small divergence from the regression line and indicated that the LD5 fraction was over 59%, which means that the M subunit was dominant in the five samples. The normal LD activity range was calculated as 108–202 U/L. The results showed that L-type Wako LD·IF reagent has sufficient performance for routine testing.
The alkaline phosphatase (ALP) activity test was converted from the JSCC method to the IFCC method on 1 April 2020. We conducted a basic evaluation of ALP activity using “L-type Wako ALP·IFCC” manufactured by Fujifilm-Wako. Parallel-run precision and reproducibility for all control materials were good with a C.V. below 1.4%. The detection limit determined from dilution linearity using 10 dilution series of high-activity ALP samples was 1,762 U/L. There was no interference by bilirubin F, bilirubin C, chyle or hemoglobin. The correlation between currently used reagents was good with the correlation factor r = 0.990, but the correlation factor for IFCC reagent result was 1/3 lower, as determined with the regression equation y = 0.344x + 0.58. ALP isozyme analyses of seven samples using “Quick Gel ALP” manufactured by Helena showed a small divergence from the regression line and indicated that the ALP5 fraction was dominant (over 57%). The normal ALP activity range was calculated as 34–106 U/L. The results indicate that L-type Wako ALP·IFCC reagent has sufficient performance for routine testing.
Rapid diagnostic kits are widely used as diagnostic tools for influenza virus infections, but false negatives are also obtained in the initial stage of infection and because of human errors. The aim of this study was to evaluate the usefulness of SPOTCHEM FLORA SF-5520 (SPOTCHEM FLORA) in the laboratory. The subjects were evaluated by 17 (33 ± 16 years of age) medical technicians of Kagoshima Medical Center. Immunoace FluAB (Towns) was used for visual judgment, and SPOTCHEM FLORA (Arkray) was used for densitometry judgment. Each dilution series was prepared from inactivated virus samples. The detection sensitivity, variation in the result judgment, and responses to the questionnaire were compared. Positive detection was achieved with 16-fold dilution for Immunoace and 64-fold dilution for SPOTCHEM FLORA. SPOTCHEM FLORA showed no variation in the judgment, whereas Immunoace showed positive judgment rates of 23.5% for type A (0% confident) and 64.7% for type B (47.1% confident). The results of analysis of the responses to the questionnaire confirmed the merits of SPOTCHEM FLORA, such as confident judgment, reduction of input errors, and examination in parallel with other work. The introduction of SPOTCHEM FLORA not only increased sensitivity and standardized criteria for influenza tests, but also improved work efficiency, such as parallel execution with other work.
DESIGN-R is used in the evaluation of bedsore in our hospital. In addition to this evaluation, we verified whether sonography and thermography are useful for bedsore evaluation. We originally devised an echo scoring system on the basis of the literature. Patients in whom a bedsore was found underwent sonography and thermography once a week, and the changes in the DESIGN-R score and echo score over time were determined in correlation with the bedsore size, which was an objective evaluation factor. The DESIGN-R score and echo score were compared among patients and verified. In 6 of 11 patients, the change in the DESIGN-R score over time correlated with bedsore size, and in 8 of 11 patients, the change in the echo score over time also correlated with bedsore size. By thermography, we found that in 4 of 11 patients, the temperature of the bedsore site was higher than that of the normal site. By sonography, we found that in 7 of 11 patients, the temperature of the bedsore site was lower than that of the normal site. The absence of a correlation between the bedsore size and the echo score seemed to affect the scoring, that is, the setting conditions of the sonography were not unified. It is considered that thermography can be used to evaluate inflammation objectively. From the above, we considered that sonography and thermography are useful for the evaluation of the progress of bedsore, in addition to using the DESIGN-R score.
Even when no change in the skin surface is recognized, sonography and thermography can sensitively detect changes in the structures of areas under the skin. Once a week, we performed sonography and thermography of the sacrum and right and left greater trochanter parts and the right and left heels of patients suspected of being at risk of bedsore. We compared the revised echo score system that we originally devised, and we also compared the temperature difference between the normal and abnormal parts measured by thermography. We investigated whether these methods could be used to predict the risk of bedsore. As a result, in an echo image, the muscle tissue structure of the heels was not distinct in all patients, whereas the sacrum and greater trochanter part were distinct in 10 of 18 patients. In thermography, the temperature decrease to the abnormal point was observed in 20 out of 30 patients. When the echo score was high, the temperature decrease of the abnormal point was observed by thermography. Bedsores are considered to more likely to occur after abnormal bedsore measures are detected. Even in a patient who has not yet developed a bedsore, a change in the muscle tissue structure and the temperature decrease of the abnormal point could be observed; thus, sonography and thermography seem useful for bedsore risk evaluation.
We performed basic evaluation studies of ‘STACIA MEBLuxTM TEST mitochondria M2’ (STACIA), a reagent for anti-mitochondrial M2 antibody measurement by chemiluminescent enzyme immunoassay. Results showed that the CVs of within-run and between-day precisions were less than 6.0%. No effects from interfering substances (bilirubin F, bilirubin C, hemoglobin and chyle) were detected. However, the rheumatoid factor showed negative effects. In addition, the correlation of STACIA with ‘Elia mitochondria M2’ (ELIa), a comparison product, was good (y = 0.815x + 0.691, r = 0.830, n = 184). The correlation between STACIA (< Index 7.0) and ELIa was shown by the correlation factor r = 0.482 and the regression equation y = 0.071x + 0.929 (n = 128). Nineteen samples were positive for the antibody in the measurement using ELIa and nineteen samples were negative in the measurement using STACIA. Positive and negative concordance rates were good (81.5%). However, the positive concordance rate (73.2%) was lower than the negative concordance rate (98.0%). STACIA showed basic performance good enough for routine use. However, the interpretation of the data seemed to require consideration of the clinical background.
Although matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has proven its high accuracy and rapidity for microorganism identification, the differentiation between Klebsiella oxytoca and Raoultella spp. by MALDI-TOF MS is difficult because of their highly similar mass spectra. Thus, the aim of this study was to evaluate the capability of MALDI Biotyper (Bruker Daltonics) to identify K. oxytoca and Raoultella spp. using biochemical tests. We tested a collection of 124 clinical isolates previously identified as K. oxytoca or Raoultella spp. by MALDI Biotyper and two ATCC strains. MALDI Biotyper correctly identified 100% of K. oxytoca isolates to the species level. For Raoultella spp., 100% of their isolates were identified to the genus level and only 8 (23.5%) isolates were identified to the species level. The use of additional phenotypic tests is recommended for the reliable identification of this genus.
The serum-soluble interleukin-2 receptor has been used as a diagnostic aid for malignant lymphoma and as an indicator of treatment follow-up. In this study, we compared the performance of reagents by chemiluminescent enzyme immunoassay (CLEIA; Lumipulse Presto IL-2R, manufactured by Fujirebio Co., Ltd.) and latex turbidimetric immunoassay (Nanopia IL-2R, manufactured by Sekisui Medical Co., Ltd.). Furthermore, we compared the correlation of CLEIA using these reagents with the outsourced assay (Determiner CL IL-2R, Hitachi Chemical Diagnostics Systems Co., Ltd.) and evaluated the suitability of these reagents in in-hospital measurement. The performance of both reagents was satisfactory and sufficient for daily measurements. However, since Nanopia IL-2R showed nonspecific reactions in some samples, it is desirable to use it considering the occurrence of nonspecific reactions. It is possible to immediately select the appropriate treatment method and expect a clinical diagnostic contribution by reporting the results on the same day.
Primary aldosteronism (PA) is the most frequent cause of secondary hypertension, and PA could result in organ injuries. Therefore, it is necessary to diagnose PA in individuals with hypertension. The ratio of aldosterone to renin is used for the screening for PA. However, since the measurements of aldosterone and renin levels are usually performed by an RIA method, they are not commonly performed in hospital laboratories. In this study, we evaluated the basic performances and the possible usefulness of Accuraseed, which is a fully automated chemiluminescent enzyme immunoassay machine (from FUJIFILM Wako Pure Chemical Corporation), for ARC and aldosterone measurements. We obtained favorable results of within-run and between-day precision tests, linearity tests, and interference tests. We also observed good correlations between measurements using Accuraseed and other methods (an EIA method for ARC and an RIA method for aldosterone). However, we should carefully interpret the results for renin because measurement tools are different between ARC and plasma renin activity (PRA).
Background: Platelet light transmission aggregometry (LTA) can be performed using a Sysmex CS series, an automatic measuring device. However, it is observed that the sample of platelet-rich plasma (PRP) moves to the left and stays for a long time in the device owing to the multisample processing. We investigated the time course of platelet activation following platelet-rich plasma preparations on CS-2000i. Methods: PRP samples from normal healthy donors were allowed to stand from 0 to 180 min after preparation. The platelet activity of these samples with or without mixing was assessed by LTA using CS-2000i. Final concentrations of 0.5 and 2 μM ADP and 0.5 and 2 mg/μL collagen were used. We also determined the platelet count at the supernatant or lower part of the sample. Results and Conclusions: The results showed that the time course of platelet aggregation was not affected within 120 min. However, the platelet activation was attenuated when the sample was weakly stimulated with ADP after 30–90 min of platelet preparation, or was stirred before the measurement after allowing the sample to stand for a long time. Although the platelet count at the lower part of the sample was approximately 10% higher than that at the supernatant, no considerable effect on platelet activation was observed. Therefore, it was suggested that leaving the sample for a long time does not affect the results of this assay under the given conditions. However, it is necessary to pay attention to the timing of the assay after PRP preparation in the determination of platelet activation.
We have already reported 24 fluorescent substances that we identified in urinary supernatant. Some fluorescent crystals and stones (e.g., sodium urate, ammonium urate, and uric acid), and mucopolysaccharide-like substances, plant fibers, and pollen are present in urinary sediments. By fluorescence spectroscopy and thin-layer chromatography, we have additionally identified 4–7 fluorescent substances in butanol extract from sodium urate and uric acid crystals and stones. The fluorescent substances in the crystals and stones may be incorporated during the formation of the crystals and stones within the urinary tract.
Recently, there have been reports of falsely high levels of hemoglobin A1c (HbA1c) when measured using the enzymatic HbA1c assay reagent Norudia N HbA1C in patients with low catalase activity. We performed an analytical investigation of an improved reagent, (RE) Norudia N HbA1c, which has been developed to lower the rate of obtaining falsely high levels, using an EV800 Clinical Analyzer. For within-run reproducibility, the coefficient of variation was from 0.42% to 0.68%. NGSP exhibited a linear dilution from 3.16% to 17.64%. The accuracy was from 98.4% to 101.4%. On-board stability was favorable up to 28 days. HbA1c levels were not affected by free bilirubin (up to 50 mg/dL), conjugated bilirubin (up to 50 mg/dL), formazin turbidity (up to 3,000 FTU), or ascorbic acid (up to 50 mg/dL). Correlation of (RE) Norudia N HbA1c with Norudia N was high, with a correlation coefficient of 0.998 (n = 56). The regression equation was y = 1.000x + 0.010, and no extremely large deviations among samples were found. From the results presented above, the analytical performance of (RE) Norudia N was found to be favorable. The two samples that yielded falsely high HbA1c levels measured using Norudia N did not reveal any significant difference in HbA1c level between (RE) Norudia N and other enzymatic assay reagents, suggesting that (RE) Norudia N suppressed falsely high HbA1c levels. The analytical performance of (RE) Norudia N is favorable, and it lowers the rate of obtaining falsely high HbA1c levels, making it useful for routine testing.
Death caused by arteriosclerotic diseases accounts for 24% of mortality cases according to the statistics of Japan. To prevent the development of arteriosclerosis, screening for the early detection of the deterioration of vascular endothelial function, which occurs at the early stage of arteriosclerosis, with high accuracy and efficiency is important. There have been many reports on the evaluation of vascular endothelial function in patients with lifestyle-related diseases and coronary artery disease. However, no survey of healthcare workers who work daily at medical institutions has been reported yet. In this study, we evaluated the vascular endothelial function in healthcare workers by blood-flow-dependent vasodilator response (FMD) tests. We then analyzed the relationship between the FMD test results and the blood data, physical examination findings, or living environmental factors. Twenty seven of 74 subjects showed decreased vascular endothelial function (decreased %FMD value). In the analysis of influencing factors in those individuals, previously reported factors, such as age, blood pressure, blood glucose level, and smoking, were also found to be associated with vascular endothelial dysfunction in our study. In addition, low serum levels of HDL cholesterol were shown to be associated with decreased %FMD values even when they were within the normal range, suggesting that these tests may be useful for the early diagnosis of vascular endothelial dysfunction. To reduce the incidence of atherosclerotic diseases, it is important to diagnose through a screening for those influencing factors and intervene early to prevent vascular endothelial dysfunction.
In recent years, several guidelines have recommended tissue fixation with 10% neutral buffered formalin solution (10% NBFS) to ensure the quality of companion diagnosis and specimens used for cancer genome medicine. When the Japanese Association of Medical Technologists (JAMT) conducted photo surveillances of histopathological examinations, JAMT has also been conducting questionnaire surveys and education since 2015 in order to standardize the procedures of tissue fixation in Japan. The adoption rate of 10% NBFS increased over time from 38.5% (416/1,081 pts) to 80.0% (902/1,127 pts) for biopsy specimens and from 31.6% (342/1,081 pts) to 72.1% (805/1,116 pts) for surgical specimens from the start of the survey to 2019. The facilities that refused to adopt 10% NBFS cited its little use in experiments and poor fixation capability as their reasons. On the other hand, the survey results suggest that the facilities that adopted 10% NBFS are actively responding to companion diagnosis. In the survey of tissue fixation time in 2017, most facilities completely fixed both biopsy and surgical specimens within 72 hours. The facilities that completed fixation with 10% NBFS within 48 hours were 59.6% for biopsy specimens and 41.9% for surgical specimens. In the future, we should improve the fixation procedure for the standardization of histopathological examination and the qualitative preservation of specimens, so that no patients will be disadvantaged in any region.
Lupus anticoagulant-hypothrombinemia syndrome (LAHPS) is characterized by positivity for the lupus anticoagulant (LA) accompanying hypoprothrombinemia and bleeding symptoms. Here, we report a case of LAHPS diagnosed by the activated partial thromboplastin time (APTT) mixing test. A 2-year-old girl presented with subcutaneous hemorrhage after developing tonsillitis. Since the prolongation of prothrombin time (PT) and APTT was pointed out in the coagulation tests, she was referred to our hospital. The APTT mixing test was carried out to investigate the cause of the prolongation of PT and APTT. As a result, the immediate-type and delayed-type waveforms exhibited similar convex upward patterns, which suggested the presence of LA. Additional tests confirmed the positivity for LA and low factor II activity (4.1%). Because she was negative for antinuclear antibodies, she was considered unlikely to have an autoimmune disease. On the basis of these results and her history of tonsillitis, the patient was diagnosed as having LAHPS that developed concomitantly with an infection. Therefore, we again realized that the APTT mixing test was useful for rapidly differentiating the causes of clotting time prolongation.
A male patient in his forties with disseminated gonococcal infection presented to our hospital with the complaint of joint pain after sexual activity. The results of blood tests indicated inflammation, and Neisseriagonorrhoeae was isolated from a blood culture. We were unable to distinguish N. gonorrhoeae from Neisseria meningitidis solely on the basis of good growth on sheep blood agar after incubation for 24 h at 35°C in an atmosphere containing 5% CO2 or on a positive result of a rapid antigen test using the N. meningitidis Y/W135 reagent of the PASTOREX Meningitis kit. We examined the immunological reactions of N. meningitidis using the PASTOREX Meningitis kit and evaluated the growth of seven N. gonorrhoeae and two N. meningitidis clinical isolates on sheep blood agar and chocolate agar. There was no difference in the growth on the two types of solid medium. However, four of the seven N. gonorrhoeae isolates showed weak agglutination with a sensitized latex reagent specific for N. meningitidis Y/W135. In conclusion, N. gonorrhoeae and N. meningitidis can be distinguished on the basis of the results of biochemical tests and/or mass spectrometry.
Erythopoietic protoporphyria (EPP) is an incomplete autosomal dominant disorder caused by an accumulation of protoporphyrin IX (PpIX) due to an impaired activity of ferrochelatase (FECH) in the heme synthetic pathway. EPP is often found in association with photosensitivity. Blood tests including an erythrocyte photohemolysis test (EPT) and an erythrocyte fluorescence test (EFT) were performed on a 10-year-old male with photosensitivity. In the patient’s erythrocytes, we observed hemolysis induced by sunlight in EPT and autofluorescence in EFT. Porphyrin analysis showed an increased level of protoporphyrin in erythrocytes, whereas the levels of other porphyrins were within normal limits. Hence, he was diagnosed as having EPP. Genetic analysis was performed and a point mutation was found in exon 6 of the FECH gene (c.683C>T, p.Pro228Leu) in heterozygotes, and a genetic polymorphism IVS3-48C was also identified on the contralateral side of the mutant allele. Taken together, we believe that in-house EPT and EFT should be performed, which could contribute to the early diagnosis of EPP.
We report a case of hairy cell leukemia (HCL), a rare tumor diagnosed on preoperative examination, in a 48-year-old male. The patient was diagnosed as having sinusitis by a previous physician; however, he did not improve with the treatment administered. The subsequent diagnosis of septal curvature necessitated surgery, and preoperative evaluation was performed. Blood test results showed an increased white blood cell count (14.7 × 109/L), and peripheral blood smear examination revealed cells with a large bright cytoplasm containing round-to-oval nuclei, and 79.0% of nuclear chromatin showed slightly aggregated lymphocytes. Chronic lymphocytic leukemia (CLL) was suspected; however, immunohistochemical analysis of cell surface markers revealed immunopositivity for CD19, CD20, and CD22 and immunonegativity for CD5 and CD23. Naturally dried peripheral blood smears showed cells with cytoplasmic hairy protrusions (hairy cells, HCs), and further surface marker analysis revealed CD11c immunopositivity, leading to the final diagnosis of HCL. Accurate diagnosis of HCL depends on comprehensive analysis, including evaluation of cell morphology, special staining procedures, cell surface marker analysis, and genetic testing. Patients suspected of having HCL in whom other mature B-cell lymphoproliferative disorders such as CLL are excluded should undergo examination of their naturally dried peripheral smear for early confirmation and diagnosis of HCL.
Anaplastic carcinoma (AC) is an aggressive tumor of the pancreas. Herein, we report the imprint cytology of an autopsy case of pancreatic invasive ductal carcinoma with a component of anaplastic carcinoma of the pleomorphic type (AC-P). A man in his 70s was admitted to our hospital owing to rapidly elevated levels of hemoglobin A1c. Computed tomography revealed a mass in the body of the pancreas and multiple hepatic masses. The patient died of pancreatic cancer after eight months and autopsy was performed. A well-defined mass of 25 mm diameter was noted in the body of the pancreas. Imprint cytology showed large pleomorphic atypical cells occurring both singly and in clusters, and isolated atypical multinucleated giant cells. Moreover, adenocarcinoma and squamous cell carcinoma were observed. Histopathology of the pancreatic mass showed adenocarcinoma admixed with squamous cell carcinoma and pleomorphic neoplastic cells with the same morphology as those in imprint cytology specimens. The diagnosis was invasive ductal carcinoma with a component of AC-P. The multiple hepatic masses were histopathologically diagnosed as metastases of pancreatic cancer. In this case, the imprint cytology showed the typical cytomorphology of AC-P.
A working group for the standardization of puncture fluid established by the Japanese Association of Medical Technologists has compiled the methods of cytometry, cytotaxonomy, and reporting for the puncture fluid test. First, cells were counted, except red blood cells. Next, we employed the Samson classification method and Giemsa staining for the cytotaxonomic classification of three cell types. The Samson classification method classified cells into polymorphonuclear cells, lymphocytes, and other cells. Giemsa staining classified cells into neutrophils, lymphocytes, and other cells, and the percentage of each type of cell was shown. If the details of other cells were known, they would be recorded as supplemental information. The reports based on the results of these classification methods may indicate acute inflammation, chronic inflammation, or leaky pathology. They were also useful for primary screening and for determining the next tests, such as microbiology tests and cytology.