Since ultrasonography equipment was fully digitalized, the resolution performance has considerably improved, and examination techniques, including elastography, contrast-enhanced ultrasound, fusion imaging and speckle tracking imaging, have been developed. The examination has been applied to various clinical settings from general checkups and close examinations to aiding medical procedures, proving its usability in emergencies and patient bedsides as well as examination rooms. Ultrasonography requires diversified comprehension and techniques, but in most cases, it highly depends on the abilities of lab technicians. The Japan Society of Ultrasonics in Medicine has launched a system to certify skillful technicians, which makes them highly motivated to play active roles in clinical medicine. In the future, new examination techniques and methods are expected to be developed, and we, the technicians, must continue our efforts in the quest for new knowledge.
To address the needs and problems of plate media for the identification of fermentative, nonfermentative, and pure cultures of Gram-negative bacilli, we have prepared an experimental medium. The medium was prepared by dissolving 11 g of agar, 5 g of NaCl, 8 g of Gelysate peptone, 0.5 g of heart infusion broth, and 0.5 g of liver broth in 970 mL of purified water while heating. Then, 2.1 mL of 41.7% dipotassium hydrogen phosphate solution and 10 mL of 0.2% phenol red solution were added, and the resultant mixture was autoclaved at 115ºC for 15 min. When the medium temperature reached approximately 50ºC, a sterile cysteine/sugar solution was added. The composition of the cysteine/sugar solution was as follows: 0.2 g of L-cysteine, 1.5 g of D-glucose, 0.5 g of sucrose, 0.5 g of D(−)-mannitol, 0.25 g of L(+)-arabinose, and 0.25 g of D(−)-sorbitol, in a total volume of 20 mL. After stirring well, 20 mL of the medium was dispensed in a petri dish for use. In the observation of acid production from growing colonies, a satisfactory test result was obtained for pure cultures and the identification of fermentative and nonfermentative bacteria.
Because the strips used for the urine bilirubin test by the diazo coupling method often show a false positive result, confirmatory examinations of bilirubin true positivity are necessary. To determine urine bilirubin, there are the Rosin method, the Harrison method, and the Watson-Howkinson method, which are based on oxidation reaction. We evaluated these methods, including the procedures of the Harrison and Watson-Howkinson methods, to determine the most suitable confirmation method. As a result, we chose the Watson-Howkinson method (W method-C). In the W method-C, the preservation of the absorption pad was easy, and the procedure was simple, easy, and required a short time and a small sample volume. Then, we examined the W method-C in terms of the absorption pad to be used, the smallest sample volume, the detection limit, and the coincidence rate with Ictotest. We concluded that One-Shot Dry was the most suitable absorption pad, because the color that developed was easy to judge with only three drops of urine. The rate of agreement with Ictotest was good at 92.6%. The W method-C is also cheap, and thus it can sufficiently be used as a confirmation test of urine bilirubin on a routine basis.
Ammonium acid urate (AAU) crystals in urine sediment have recently started to attract attention for the possibility of calculus formation. Here, we report on our experience of treating four pediatric cases of nephrotic syndrome (NS) with AAU crystals in urine sediment. The patients, who were all boys aged 1, 4, 7, and 11 years, were hospitalized for edema and severe proteinuria. They were discharged after achieving remission with steroid therapy. In all the four patients, concentrated urine with high urine specific gravity and low urine output was noted. AAU crystals were detected in the urine only once or twice during the hospitalization. In one of the patients, AAU calculi stuck at the balloon catheter tip were noted on the same day as when the patient was admitted. Many of the mechanisms of calculus formation are unknown. In cases of pediatric NS, however, severe dehydration and urine concentration were considered to develop even without diarrhea or vomiting observed in rotavirus enterocolitis and lead to AAU crystal formation. Crystals in the urine, which indicate the saturation of components, are an important finding indicative of a risk of the onset of acute postrenal failure caused by AAU calculi in pediatric NS. In all of the cases, AAU crystals appeared during the period of severe NS. However, the longitudinal data on protein concentration, specific gravity, and volume of urine suggest that the crystals tended to appear immediately before the recovery from the disease.
Neurocutaneous melanosis is a rare congenital disease that is characterized by the presence of giant pigmented nevus of the skin and proliferation of melanocytes producing melanin in the central nervous system. We describe a case of neurocutaneous melanosis, in which we detected cells in the cerebrospinal fluid showing abnormal Samson staining. The diagnosis was confirmed by the detection of abnormal cells that have pigment granules by cytological analysis of the cerebrospinal fluid. We were able to detect pigment granule cells not only by cytological analysis but also by Samson staining of the cerebrospinal fluid. This case shows that screening by Samson staining can provide clinically important information in terms of the diagnosis of neurocutaneous melanosis.
We report here a rare case of blastic plasmacytoid dendritic cell neoplasm (BPDCN). Tumor cells in this case had a weakly stained basophilic cytosol and nucleus with fine nucleoreticulum and plural numbers of nucleoli. However, pseudopodia-shaped cytoplasmic expansions and vacuoles along the cytoplasmic outline, which are specifically seen in BPDCN cells, were not identified. In BPDCN, various patterns of cell morphology are observed. Similarly to the current case, especially in cases showing blastic cells without morphologic features specific to BPDCN cells that are observed in peripheral blood or bone marrow, it is extremely difficult not only to speculate BPDCN but also to perform differential diagnosis with other types of acute leukemia and leukemic changes of lymphomas by May-Giemsa staining. Many cases do not show typical expression patterns of cell surface antigens (CD markers), and the diagnosis cannot be obtained unless plasmacytoid-dendritic-cell-related CD markers such as CD123, which was confirmed in the current case, are examined. Moreover, rearrangements of the T cell receptor and immunoglobulin genes and of the Epstein-Barr virus (EBV)-encoded RNA 1 (EBER-1) should be negative for a rapid and definite diagnosis. BPDCN is rare but the prognosis is extremely poor. The patient described in this report died at 6 months after the diagnosis. To perform therapy at the early stages, it is important that medical technologists must assume that BPDCN is the disease.
Endoscopic ultrasonography fine needle aspiration (EUS-FNA) cytology has proven to be useful for the evaluation of pancreatic tumors or gastric submucosal tumors. In the present study, utilizing the cell block procedure, we examined a diagnostic approach of endoscopic ultrasonography fine needle aspiration (EUS-FNA) cytology in two patients with these lesions. In patient 1, EUS-FNA cytology showed round cells in loose small clumps with focal rosettes or a ribbonlike structure. Immunohistostaining by the cell block procedure revealed the positivity for chromogranin A and synaptophysin in the tumor cells. We diagnosed a neuroendocrine tumor. In patient 2, EUS-FNA cytology showed spindle tumor cells with nuclear palisading. Immunohistostaining by the cell block procedure revealed the positivity for DOG-1, c-kit, and CD34 in the tumor cells. We diagnosed a gastrointestinal stromal tumor. Our findings suggest that EUS-FNA cytology with the cell block procedure offers diagnostic efficacy in patients with pancreatic tumors or gastrointestinal submucosal tumors.
Tympanometry is a method of measuring the compliance of the tympanic membrane and the middle ear pressure of the eardrum and of diagnosing the existence of middle ear storage liquid. Tympanometry using a probe sound of 226 Hz has generally been used, but it shows an A-type tympanogram in neonates with a soft external auditory canal even if there is middle ear storage liquid, owing to a change in pressure. On the other hand, there is a report showing that tympanometry using a probe sound of 1,000 Hz is more reliable. In this study, we compared and examined the differences between the impedance audiometers RS-22 and GSI Tympstar Version 2 with probe sounds of 1,000 and 678 Hz in our institution. Eleven ears showed the A type when using a probe sound of 226 Hz and the B type when using probe sounds of 1,000 and 678 Hz. All 11 ears were those of babies less than twelve months of age. Our results indicated that tympanometry with probe sounds of 1,000 and 678 Hz is useful for babies less than 6 months of age. The probe sound of 1,000 Hz showed artifact-like waveforms and reexamination is needed. GSI requires time for apparatus operation because it is of the syringe type. These results indicated that its measurement was difficult for children who cannot keep still. An audiometer with the structure and function that can fix an earplug and have the probe sound of 1,000 Hz and with the pump type is required.
Over 18 months between October 2012 and March 2014, in 168 patients with a violent or lingering cough that needed to be differentiated from pertussis, a laboratory diagnostic tests of pertussis was performed retrospectively. To detect the pathogen, postnasal swabs were collected from all the patients for the isolation of Bordetella pertussis and the detection of DNA using the loop-mediated isothermal amplification (LAMP) assay. For serodiagnosis, the PT-IgG titer was measured in 126 patients (67 sets of paired sera and 59 single serum samples). Diagnosis of pertussis was defined as the isolation or DNA detection of B. pertussis or meeting the criteria for serodiagnosis. The diagnosis of pertussis based on laboratory test findings was made in 34 of the 168 patients (20%). The median age at their first visit was 0.9 years (17 days to 12.3 years). DNA was detected in 16 patients (47%), and B. pertussis was isolated in 9 (26%), all of whom were positive for DNA. Thirty-one patients (91%) met the serodiagnosis criteria, 18 (53%) of whom only falsely met these criteria. Among 7 of the 18 patients, who were unvaccinated young infants (aged 17 days to 3 months), 6 showed a decrease in the antibody titer in paired sera and one vomited after coughing caused by pyloric stenosis. These 7 patients were considered to have falsely met the serodiagnosis criteria owing to the transplacental transfer of maternal antibodies. Pathogen detection is more reliable for the diagnosis of pertussis based on laboratory test findings, with the LAMP assay being convenient, rapid, and sensitive for this purpose.
Urinary C-peptide examination is used as an indirect method of assessing the level of insulin production. We use 24-h urine samples as materials for examination. We use a Na2CO3-containing stabilizer of urinary C-peptide. Under acidic conditions, C-peptide is unstable and is decomposed by growing bacteria. However, the effect of the concentration of the Na2CO3-containing stabilizer on the measurements of collected urine volume is unknown. We examined the effects of the concentration of the Na2CO3-containing stabilizer, storage temperature, and bacterial presence on the measurements of collected urine volume. According to our results, CPR was decomposed by growing bacteria, and the urinary CPR concentration decreased when the collected urine volume per package of the stabilizer exceeded 4,000 mL at room temperature. When we chilled and preserved urine with low concentrations of the stabilizer, CPR was stable. In addition, in the case of the collected urine volume that was less than 300 mL per package of the stabilizer, the urine CPR concentration decreased. After examining the effects of both pH and salt concentration, we considered that salt concentration has a greater effect than pH on CPR when a high concentration of the stabilizer is used. Therefore, we must determine the appropriate concentration of the Na2CO3-containing stabilizer for a particular collected urine volume before measurements.
The activated partial thromboplastin time (APTT) cross-mixing test is useful for screening coagulation factor deficiencies and inhibitors including lupus anticoagulants. Because the mixing test procedure is complicated owing to the manual preparation of specimens, the test is performed in only a few laboratories. Here, we used the fully automatic clinical laboratory system STACIA for mixing test, which is a general-purpose machine (LSI Medience, Inc.) and we deviced a more easily mixing test method as the SATCIA method. In addition, we compared the STACIA method with the TOP method which is the semiautomatic mixing test method using ACL-TOP500 (Instrumentation Laboratory, Inc.). The TOP method requires the manual preparation of a diluted sample for warming, but in the STACIA method, this is performed automatically. So the STACIA method is simple and easy, because dispensing work is not necessary. Also, we calculated the sensitivity for detecting inhibitors by the waveform pattern method, ICA, and CMT index, respectively for evaluating the STACIA method. The sensitivity values observed in the waveform pattern method, ICA, and CMT index were 75.0%, 79.2%, and 91.7%, respectively. The CMT index was the most sensitivity is good. The STACIA method can be measured both of immediate test and incubated test with half the sample volume required in the TOP method. Also, this method leads to both labor saving and increased accuracy of the results. In addition, numerical values were obtained for the evaluation of the results by this method. We conclude that the mixing test using STACIA can be useful for detecting coagulation abnormalities such as factor deficiencies or inhibitors as part of routine duties.
CicaFit AMY-G7 is a reagent kit that was designed as a transferable method proposed by the Japan Society of Clinical Chemistry (JSCC) for the measurement of amylase (AMY) using 4,6-ethylidene-4-nitrophenyl-α-D-maltoheptaoside (Et-G7-PNP) as a substrate. The values of AMY activity in patient serum and urine measured using CicaFit AMY-G7 were compared with those obtained by the Standard Operating Procedure for the measurement of the catalytic concentration of AMY established by the Japanese Committee for Clinical Laboratory Standards (JCCLS-SOP). The results obtained using CicaFit AMY-G7 were in good agreement with those determined by the JCCLS-SOP, regardless of variations in the proportion of pancreatic and salivary isoenzymes (P/T%) in each sample or sample type (serum or urine). For assays using α-2-chloro-4-nitrophenyl-β-D-galactopyranosylmaltoside (Gal-G2-CNP) and 2-chloro-4-nitrophenyl-α-D-maltotrioside (G3-CNP) as substrates, correlation analysis revealed a marked difference in the slopes of regression lines between serum and urine samples. Other kits employing the same substrate, namely, Et-G7-PNP, tended to give higher values for serum samples. The values of AMY activity in external quality assessment (EQA) samples measured using other kits differed from those obtained by the JCCLS-SOP when recombinant human AMY was added to the samples. Other kits using the Et-G7-PNP substrate gave slightly higher values and those using the G3-CNP substrate provided slightly lower values (within ± 5% of the target value for both), while those using the Gal-G2-CNP substrate gave values that were below the permissible lower limit.
We devised a new cell block (CB) manufacturing method called the ‘paraffin-agar sandwich method’ and then compared it with conventional CB manufacturing methods such as the ‘fibrin clot method’, ‘agar method’, and ‘sodium alginate method’. The purpose of this study was to investigate the cellular collections and staining conditions of body fluid cavity specimens that were processed using the four CB manufacturing methods. Our results showed that among the four CB manufacturing methods, the paraffin-agar sandwich and agar methods provided significantly larger numbers of cells derived from cell pellets of 10 or 50 μL volume than the fibrin clot method. When cell pellets of 100 μL volume were used, the paraffin-agar sandwich method provided significantly larger numbers of cells than both the fibrin clot and sodium alginate methods. The investigation of the staining conditions in the different preparation backgrounds showed more contamination of the preparation background in both the fibrin clot and sodium alginate methods than in the other methods. Therefore, it was shown that the paraffin-agar sandwich method is an improvement over the other CB methods for cellular collection and that it may be used for various purposes.
Hepatitis B virus (HBV) reactivation may occur not only in hepatitis B surface antigen (HBsAg)-positive patients but also in patients with resolved HBV infection who are seronegative for HBsAg but seropositive for the anti-hepatitis B core antibody (HBcAb) and/or anti-hepatitis B surface antibody (HBsAb). The detection of HbcAb is important before chemotherapy. In this study, we compared the sensitivity and specificity of HBcAb in 77 low-titer samples with nearly 1.0 cut-off index (C.O.I) using 3 kits for the detection of HBcAb, namely, HISCL [chemiluminescent enzyme immunoassay (CLEIA)], Presto (CLEIA) and Architect [chemiluminescent immunoassay (CLIA)]. The rate of agreement between Presto and Architect was 96.1% owing to the use of the same hepatitis B core antigen (HbcAg)-anti-human IgG antibody sandwich assay. However, because only HISCL was applied to the HBcAg-HBcAg sandwich assay, the rates of agreement between HISCL and Presto and HISCL and Architect were 79.2% and 80.5%, respectively. As shown by additional information, each assay had false-positive and -negative data, suggesting that it is difficult to determine which of the three is better, particularly in low-titer samples. To detect the low titers of HBcAb, a more sensitive HBcAb assay or a highly sensitive HBsAg assay is required to manage HBV reactivation.
Particularly in Japan, the number of individuals affected by cedar pollen allergy is increasing every year owing to continuous mass scattering of pollens and similar allergens. In the future, the number of affected individuals will increase and younger patients are expected. Since pollen dissemination is considerably affected by the terrain and climate change, information on the region can be used to establish antigen avoidance measures and prevent pollen exposure. Furthermore, providing pollen information together with the results of specific IgE antibody tests can help improve the quality of life (QOL) of sufferers. Thus, in order to grasp the actual cedar pollen situation, we performed measurements of airborne pollen count from 2012 to 2014 and considered the relationship between the contract number and the rate of positivity for the cedar-specific antibody, as determined by the tests conducted in the Institute. Pollen count has been suggested to affect the contract number and positivity rate. Thus, it is important to take advantage of research and information specific to the region.
Mycoplasma pneumoniae (Mp) is a major pathogen causing pneumonia in children. We have used the loop-mediated isothermal amplification (LAMP) method to detect Mp DNA from patients, and here, we report our results obtained over the last 5 years. From April 2009 to March 2014, we collected pharyngeal swabs from 2,215 children who were hospitalized with pneumonia, and performed the LAMP method to detect Mp DNA. Among the 2,215 children, 712 (32%) were positive for Mp DNA. Their ages ranged from 1 month to 15 years and 6 months, and the median age was 7 years and 2 months. Regarding the annual number of patients positive for Mp DNA, there were 37 patients in 2009, 161 in 2010, 308 in 2011, 147 in 2012, and 59 in 2013. In 2011, there was a nationwide epidemic of Mp pneumonia, and the highest number of patients with Mp pneumonia was also seen in September 2011 at our hospital. When we examined the number of patients by calendar month, there was a seasonal trend for more Mp DNA-positive patients from summer to early winter and fewer patients in spring. In all the 712 children, we measured the antibody titer in paired serum samples and confirmed seroconversion or an increase in the titer by 4-fold or more in 559 patients (79%). In conclusion, Mp DNA detection by the LAMP method can be performed simply and quickly with high sensitivity, making it a useful laboratory diagnostic method for children with Mp pneumonia.
Recently, it has become a concern that the number of young medical technologists who participate in the meetings organized by the Association of Medical Technologists has been decreasing. Therefore, an open discussion forum was established to solve this problem nationwide. As a local trial, the Nagano Association of Medical Technologists organized the “Executive Committee of Young Medical Technologists”, which was subsequently renamed the “Department of Young Medical Technologists”. This organization asked young members’ opinions about what they would like to request to the Association of Medical Technologists. Then, in accordance with their requests, the Department of Young Medical Technologists held some workshops such as “Specialized Workshops for Communication” and “Skills Training Courses” for four years. These trials achieved good results; for example, the number of academic conference presentations that involve collaboration with young members among different institutions has increased, and several independent study meetings that were planned by young members have been held. In addition, more young members participated in the different research teams of the Nagano Association of Medical Technologists. For establishing an association of young medical technologists, we should always make an effort to understand their needs and plan appropriate events. Therefore, the Association of Medical Technologists need to activity promote the position of young members in order to form an organization composed of young members.