The target patients were those with a bedsore onset (ulcer group: 17 patients) and those with non-bedsore onset (nonulcer group:15 patients). It was slight, and the OH scale used was moderate. I assumed that the standard N-L ratio is less than 5. There were many patients in whom the N-L ratio was less than 5. The nonulcer group examined showed a change in the N-L ratio every week, and the ulcer since the hospitalization of the nonulcer and ulcer groups showed significant differences in N-L ratio. Few had an N-L ratio less than 5, and there were many with an N-L ratio greater than or equal to 5, which did not show a transient increase after the following week. An N-L ratio greater than or equal to 10 was observed in the ulcer group. In addition, I found that the N-L ratio was high on the bedsore onset day and I frequently observed patients with an N-L ratio greater than or equal to 5 one week before an ulcer developed (72.7%; 8/1 patients). Ulcer prediction factors such as the OH scale score are important in determining the way that a patient is made to lie down and in patients with difficulty in moving their body themselves in order to determine the change in the N-L ratio. I was able to confirm in particular the usefulness of the OH scale from the viewpoint of ulcer prediction.
In the routine examination for urinary sediments, we often detect hyaline casts in patients under cardiovascular medication. In this study, we established five conditions for excluding patients presenting with renal dysfunction and in whom hyaline casts were also detected and analyzed the obtained data so that we can evaluate the relationship between the presence of hyaline casts and the brain natriuretic peptide (BNP) as a marker of heart failure (HF). Detection of hyaline casts from urinary sediment showed a positive correlation with that of BNP, which suggests that the presence of hyaline casts is related to cardiovascular disease. Regarding this relationship, hyaline casts formed in the kidney and BNP as a maker of HF may be associated with cardiorenal interaction. Moreover, the rate of hyaline cast detection and the number of hyaline casts gradually increase, reflecting the BNP data. It is suggested that the number of detected hyaline casts may be clinically useful as an estimate of the severity of cardiovascular disease.
It has been revealed that lipopolysaccharide, which is a component of the cell walls of Gram-negative bacteria, induces monocytes and vascular endothelium cells to produce interleukin-6 (IL-6) and tissue factor (TF), and its relevance to the development of disseminated intravascular coagulation (DIC) its symptoms has attracted attention. We investigated the relationship between the positivity rates for TF/IL-6 and bacterial strain in DIC sepsis. In this study, we used Gram-negative bacteria, Gram-positive bacteria and yeast as stimulants. Moreover, we stimulated cells with both bacterial cells and their extracts. We prepared cells of the human monocyte cell line U937 and peripheral blood mononuclear cells (PBMCs), stimulated them with stimulant LPS, various clinical bacteria [Escherichia coli (E. coli), Staphylococcus aureus, coagulase-negative Staphylococci (CNS), Klebsiella pneumoniae, Candida albicans] and their extracts. As detection procedures, we immunostained U937 cells or PBMCs with an FITC-labeled anti-human TF antibody and a PE-labeled anti-IL-6 antibody, and measured by flow cytometry the extracellular and intracellular products of TF and IL-6 before and after the addition of the stimulant. There were significant differences in positivity rates among bacterial species, whereas no significant differences were observed among bacteria strains. Furthermore, there were no significant differences in positivity rates between bacterial bodies and their extracts. In U937 cells, E. coli induced the highest positivity rate for TF, and CNS induced the highest positivity rate for IL-6. In PBMCs, there were individual differences in positivity rates for TF and IL-6. As to the reactivity of a living body with sepsis, we suggest that differences in individual specificities and sensitivities of living bodies are larger than those in bacterial species and strains.
Decreased red cell A and B antigen expression levels are observed in some hematological malignancies, including acute leukemia. Expression dynamics of I antigens that reside in the subterminal portions of oligosaccharides converted to A, B, and H antigens were analyzed by flow cytometry (FCM) to differentiate from decreased A and B antigen expression levels due to serovars or chimera. We statistically compared I antigen expression level and fluorescence intensity between hematological and nonhematological malignancies with decreased A and B antigen expression levels. Results showed that the I antigen expression level was significantly low for only hematological malignancies. Thus, analysis of I antigen by FCM is more likely to differentiate decreased red cell A and B antigen expression levels due to hematological malignancies from a congenital variant blood type.
The leading pathological causes of chronic heart failure (CHF) are hypertension and myocardial ischemia, which are intimately related to atherosclerosis. Dyslipidemia is one of the major risk factors for atherosclerosis. Therefore, we investigated the relevance of serum lipid profile to B-type natriuretic peptide (BNP) in patients with CHF. CHF patients (n=81) and healthy control subjects (n=90) were investigated. BNP was measured by enzyme immunoassay. The lipoprotein profile was analyzed by anion-exchange chromatography. The correlations between lipid profile and BNP levels in patients with CHF were studied. HDL and VLDL cholesterol levels in CHF patients were lower and higher than those in healthy control subjects, respectively. Simple correlation analysis showed that HDL, LDL, and VLDL cholesterol levels significantly and inversely correlated with BNP levels in CHF patients. Moreover, multivariate stepwise regression analysis revealed that HDL cholesterol levels significantly and inversely correlated with BNP levels. These results suggest that decreasing HDL cholesterol levels may be a major therapeutic target for atherosclerosis progression in CHF patients with dyslipidemia, but this requires further investigation.
AmpC genes are encoded on the chromosome of Enterobacter cloacae. Excessive production of AmpC β-lactamases is induced by an antimicrobial agent. We examined the excessive production of AmpC β-lactamases using the E. cloacae strain that had been isolated from a clinical specimen by two methods. Consequently, AmpC β-lactamases were detected in 3.7% of the strains by the Cica β test, which is a third-generation cephem antibiotics susceptibility test in which AmpC genes were induced by the disc diffusion method. Additionally, for induction, the cells were cultured in CAZ Sub-MIC liquid culture medium with shaking, and the results showed 80% positivity for the lactamases. It is considered that there is a possibility that strains excessively producing AmpC β-lactamases exist in vivo, even when AmpC β-lactamases are not detected by clinical isolation because it is considered that a newer method is closer to the in vivo environment in terms of conditions. Moreover, this result shows that it is very important in antibiotic therapy to follow the dosage regimen, because antibiotic therapy with insufficient dosage promotes the excessive production of AmpC β-lactamases, and the patient may acquire an intractable infectious disease.
There are reports on new parameters (NEUT-X and NEUT-Y) of neutrophils and their relationships with toxic granules in patients with inflammatory diseases. However, there is no report yet on vacuolization and on these parameter relationships. Therefore, we analyzed the relationship between the results of immunoassays and morphological changes of neutrophils in relation to NEUT-X and NEUT-Y in patients with inflammatory diseases. In this study, the patient group showed significantly higher NEUT-X and NEUT-Y levels than the control group. These levels showed a significant correlation with CRP level and PCT, but not with neutrophil count and white blood cell count. Also, toxic granules and vacuolization were reflected as cytoplasmic complexities formed by NEUT-X, and the increase in the number of immature neutrophils was reflected as an increase in the quantity of intracellular RNA induced by NEUT-Y. On the other hand, without intending to reflect bacterial infection directly, NEUT-X and NEUT-Y had captured the reactivity change of neutrophils in response to inflammation. Furthermore, in this study, the NEUT-X and NEUT-Y levels were high owing to severe sepsis. NEUT-X and NEUT-Y used as markers showed insufficient specificity and diagnostic sensitivity, but objective data can be obtained quickly and easily using an analyzer. In addition, these parameters may be helpful for diagnosing sepsis.
The monitoring of Epstein-Barr virus (EBV) DNA loads is important for the detection and prevention of EBV-related disorders. EBV loads have been evaluated by quantitative real-time PCR analysis requiring the preparation of standards. The standard materials for the quantification of EBV loads vary among clinical laboratories. Therefore, it is difficult to obtain comparable EBV DNA quantitative values among laboratories. In this study, the copy number of plasmids containing a fragment corresponding to the EBV BNRF1 gene was calibrated with the 1st WHO International Standard for EBV quantification tests, and then the calibrated plasmids were used as secondary standards to quantitate the EBV DNA loads of subsequent samples from patients. Additionally, the theoretical conversion factor was calculated to convert the EBV DNA loads obtained using noncalibrated standards into those obtained using the present secondary standards. The EBV DNA loads in hematopoietic-stem-cell-transplanted patients suspected of having post-transplant lymphoproliferative disorder (PTLD) and patients with EBV-related hemophagocytic lymphohistiocytosis (EBV-HLH) were 8.74 × 102−1.51 × 104 and 1.58 × 104−1.33 × 106 IU/μg DNA, respectively, as determined by these methods. The utilization of secondary unified standards for the quantification of EBV DNA loads available for many clinical laboratories would contribute to the establishment of a diagnostic reference and therapeutic intervention criteria based on the quantitative values of EBV DNA, not only in PTLD patients but also in those with various disorders related to EBV infection.
We report two cases of severe endometritis in which encapsulated bacillus infection was detected by endometrial cytology, and vaginal culture revealed Klebsiella pneumoniae infection. Case 1: The patient presented with leucorrhea. Encapsulated bacilli were observed in endometrial smears, and vaginal culture revealed K. pneumoniae infection. One month later, the patient was brought to an emergency hospital and diagnosed as having tuboovarian abscess and peritonitis. Case 2: The patient presented with metrorrhagia, abdominal pain, and fever. Encapsulated bacilli were observed in endocervical and endometrial smears. Bilateral oophorectomy and hysterectomy were performed following the diagnosis of acute endometritis, pyosalpingitis and multiple leiomyomas of the uterus. Usually, the aim of cytology is to diagnose the existence of malignant tumors, but not infectious diseases. However, endometrial culture has not been performed for the diagnosis of gynecologic infectious diseases. Thus, we should provide information of infection status in cytology reports.
We would like to report the case of a 69-year-old man who visited a local physician after suffering from abdominal pain and fever that lasted for a week. The patient was diagnosed as having mesenteric adenitis, and levofloxacin (LVFX) was prescribed. However, his symptoms did not improve, and the patient was admitted to our hospital two days later. The antibiotic LVFX was changed to cefmetazole, and the condition of the patient was monitored. Three days after admission, nausea, decreased blood pressure, brown urine, and yellowing of the eyeballs were observed, and significant anemia (Hb level, 6.7 g/dL) and hemolysis were observed in a blood test. On Day 4, his anemia advanced (Hb level, 4.0 g/dL) and positivity for autoantibodies was shown by the direct antiglobulin test (DAT) and indirect antiglobulin test (IAT) carried out as a pretransfusion test, and autoimmune hemolytic anemia was diagnosed. On Days 4 and 5, two units of packed red blood cells were transfused, and steroid pulse therapy was initiated on Day 5. Improvement in anemia conditions and a rapid decrease in the number of autoantibodies were observed. Negative IAT and DAT results were obtained on Days 13 and 28, respectively. A drug lymphocyte stimulation test was carried out on Day 43, which showed positivity for LVFX. In this patient, on the basis of his clinical course, drug-induced immune hemolytic anemia (DIIHA) was presumed to be induced by LVFX. Although a rare disease, there are reports of DIIHA caused by frequently used drugs such as antibiotics and antihypertensive agents. Thus, the possibility of DIIHA should be considered in connection with their use. As seen in this case, symptoms can become severe; thus, it is important that we do not overlook hemolytic anemia and keep in mind the possibility of DIIHA when DAT results are positive.
DiaPack3000, developed as the next model following DiaPack2000, is a specific IgE antibody analyzer. Its measurement time is 12 min/test, and it can perform 90 tests in 39 min. This analyzer enabled the shortening of measurement time, improvement of processing capacity compared with previous analyzers, insertion of samples at any time, and enhanced operability. We evaluated the basic performance of DiaPack3000, and compared it with those of DiaPack2000 and Phadia5000. DiaPack3000 was found to be superior to DiaPack2000 in terms of reproducibility and sensitivity. Dilution linearity was obtained throughout the measurement range. In addition, no carryover was observed. The coefficients of correlation between DiaPack3000 and DiaPack2000 were 0.882–0.999 in six items. In addition, the rates of agreement of the class with Phadia5000 were 80.7–91.9% in six items. The performance of DiaPack3000 was adequate; therefore, DiaPack3000 could be more efficient for allergy testing in routine clinical practice than previous models.
Fluorescence in situ hybridization (FISH) methods are an important technique for pathological diagnosis and the estimation of the efficacy of molecular targeted drug therapy. The HISTRA HER2 FISH kit (JOKOH) is a commercially available system by which FISH methods can be performed in a short time. We examined the conditions of heat treatment and enzyme treatment time in FISH methods using the HISTRA HER2 FISH kit in gastric cancer. Ten cases of gastric cancer were diagnosed in Mie University Hospital. The signal visibility was improved under the study conditions, namely, 40 min of heat treatment and 10 min of enzyme treatment, and 60 min of heat treatment and 10 min of enzyme treatment. Extension of the heat treatment time was considered to improve signal visibility.
A preliminary assessment was conducted before implementing the RAPIDPoint® 500 blood gas system (RP500), a maintenance-free system using cartridges, to investigate its fundamental performance. We compared RP500 with the RAPIDLab 1265 blood gas system (RL1265) in terms of the following ten parameters: pH, pCO2, pO2, tHb, and levels of Na+, K+, Cl-, Ca2+, glucose (Glu) and lactate (Lac). We also compared RP500 with the LABOSPECT 008 biochemical analyzer (LAB) in terms of the levels of Na+, K+, Cl-, and Glu, and with Dimension (Dim) in terms of Lac level. Although repeatability and intralaboratory precision were satisfactory, Lac levels tended to start increasing gradually after a cartridge was inserted into the system. RP500 results showed good correlations with RL1265 results and also showed favorable correlations for plasma samples analyzed using LAB and Dim. For this maintenance-free RP500 system, users only need to replace a wash cartridge once every ten days and a reagent cartridge once every 28 days. By using this system, users can feel much less burdened than with conventional analyzers. In addition, RP500 can be used concurrently with the Automatic Quality Control (AQC) cartridge and the RAPIDComm data management system, which enables the centralized management of blood gas analyzers that are compatible with RAPIDComm and located in different places, such as ICUs, emergency medical care centers and hospital wards, by connecting these systems on a network. RAPIDComm-connected analyzers can be monitored and remotely provided troubleshooting as required, which may help users reduce or eliminate the time for moving around in a facility and will consequently be laborsaving and contribute an efficient laboratory environment.
High-resolution sonography is capable of depicting peripheral nerves with excellent visual quality. However, it is unknown whether changes related to aging can be detected by sonography. The aim of this study was to examine the power of sonography to detect aging. For sonography, a high-frequency linear array transducer (central frequency, 12 MHz) was used. We examined the median nerve by measuring motor conduction velocity (MCV), fascicular cross-sectional area (CSA) at the levels of the wrist and elbow, and anteroposterior diameter at the levels of the forearm, lunate and capitate bones in 34 healthy subjects (12 males, 22 females; median age, 34.5 years; age range, 22−78). MCV tended to decrease with aging; however, CSA was not significantly different among age subgroups. A significant correlation between MCV and CSA was observed in the subgroups of twenties and thirties, but not in the subgroups over 40. This is probably due to the fact that CSA of the median nerve measured by sonography does not change with aging, whereas MCV decreases with aging because of nerve degeneration. In this study, sonography depicted physiological size differences of the median nerve at the carpal tunnel but not age-related changes.
This study was conducted to evaluate the basic performance of a newly developed squamous cell carcinoma (SCC) assay reagent for the AIA based on fluorescence enzyme immunoassay (FEIA). The reactivity of this reagent with SCC antigens was also evaluated and compared with actual measured values from patients with various diseases. The basic performance of the new reagent was shown to offer good precision and specificity. An evaluation of its correlation with the performance of the reagent used in the ARCHITECT, the conventional assay for SCC, revealed a number of cases with markedly different values. To identify causal factors, the reactivities of the reagents for AIA and ARCHITECT with the recombinant antigens SCCA-1 and SCCA-2 were compared, which showed them to be highly reactive with SCCA-2 and SCCA-1, respectively, suggesting that this difference in reactivity may be a cause of the markedly different values. We also encountered another case of a benign disease with deviating elevated values obtained using the ARCHITECT, in which polymerized SCC antigens resulting from the binding of IgG and SCC antigens in the blood were thought to be associated with the falsely elevated values determined using the ARCHITECT. Other evaluations revealed a high positivity rate for SCC and positivity rates equivalent to those obtained using the ARCHITECT for various other diseases. This study shows that the new SCC assay reagent for the AIA, which offers good precision and specificity and allows measurements to be taken in approximately 20 min, appears to be highly useful in routine laboratory tests, particularly those performed prior to clinical consultation.
The effects of calcium and magnesium ions on the measurement of the specific gravity of urine with a test strip were investigated. Both ions gave significantly higher values than NaCl. The salts of calcium and magnesium indicate the characteristic that both ions lower the pHs of the citrate buffer, phosphate buffer and EDTA2Na solution. From this result, it can be interpreted that calcium and magnesium ions react with substances contained in the detection system to liberate hydrogen ions, resulting in a false positivity.
The 2D code is useful in the management of clinical pathology systems. When we introduced a new pathology system, Micro QR Code printable cassette printers were installed. However, frequent recognition error by the Micro QR Code was observed on the cassettes after paraffin embedding. Here, we investigated the recognition rate by comparing the Micro QR Code with Data Matrix ECC200 under the same conditions. As a result, 14 errors were observed in 522 routine cassette samples when using the Micro QR Code, whereas only 1 error was observed when using the Data Matrix ECC200. Thus, the recognition of Data Matrix ECC200 was quicker and more accurate than that of the Micro QR Code. With the Micro QR Code, the position detection pattern had a scratch and a defect on the cassette, which were not observed when using the Data Matrix ECC200. In a pathology system, it was assumed that the position detection pattern affected the recognition rate or speed of the 2D code system. These data indicated that Data Matrix ECC200 is useful for the cassette printing in our operation, in terms of maintaining the position detection pattern and recognition speed.
A decision was made to renew an existing information system along with our facility reconstruction. The main objective of the project is twofold: shortening of inspection time and strengthening of countermeasures against inspection malpractices. We verified the process from outpatient admission to inspection completion in order to reduce the inspection time. A system was developed to improve the efficiency of specimen reception via the centralized system of blood sampling, urine collection, and physiological function test. For countermeasures against inspection malpractice, we selected a departmental system that meets required specifications, and amended the authentication process in the blood transfusion test. Since the centralized admission could not be achieved only by utilizing standard functions, a system was developed to admit patients in an all-inclusive manner by a method that allows aggregation of information requested by departments pertaining to various inspection types and master code systems. Unavoidable problems associated with specific attributes of the system were addressed by amending the operation of the system. As a result, the average time required to report on inspections was reduced from 45 to 35 min. On the other hand, the development of countermeasures against inspection malpractice requires further elaboration.
The number of end-stage kidney disease patients in Japan has exceeded 300,000, and a large amount of money is spent for dialysis. To solve these important problems, it is necessary to determine renal damage at an early stage. In this study, we used CLINITEK Novus provided by Siemens Healthcare Diagnostics, and examined the correlation between albumin and creatinine in urine detected by a dipstick method and a quantitative method. Simultaneously, we analyzed the albumin/creatinine (A/C) ratio, and then evaluated the usefulness of these findings using the renal function parameter (estimated eGFR) and chronic kidney disease (CKD) risk stratification. The results of the dipstick and quantitative methods showed good correlation, and the coincidence rates were 85.9% for albumin and 65.7% for creatinine. Moreover, the A/C ratio determined using these values also showed a good coincidence rate of 85.9% for the dipstick and quantitative methods. The A/C ratio was positive in more than 74.6% of the patients with renal dysfunction. These findings suggest that the dipstick method has potential usefulness for rapid screening.