Reference values for bisphosphoglycerate mutase protein content and specific activity in human erythrocytes.
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Adult↗
Biomedical subjects
I Tsuda
Publications and source records attributed to I Tsuda.
DNA extraction from human urinary sediment.
DNA was extracted from urinary sediments and was sufficient for polymerase chain reaction (PCR) and enzymatic analysis, even if DNA from microorganisms coexisted. From urine samples, the yield of DNA ranged from trace levels to 20 micrograms per 10 mL urine. When urinary sediment was stored in ethanol, DNA remained stable for 2 weeks or more. Individual identification and sex determination could easily be performed using either fresh or ethanol-fixed urine. In conclusion, urine can be used as a source for PCR-based investigations and genetic studies.
Adolescent↗
An enzyme-linked immunosorbent assay and reference ranges for bisphosphoglycerate mutase in human erythrocytes.
We established an enzyme-linked immunosorbent assay (ELISA) system for the determination of human bisphosphoglycerate mutase (BPGM) protein content in human erythrocytes using a polyclonal anti-BPGM antibody, we determined reference ranges for BPGM protein content, synthase activity, and specific activity in human erythrocytes. We produced a recombinant human BPGM (rBPGM) by gene manipulation using E. coli and then obtained the polyclonal antibody by immunizing rabbits with purified rBPGM. The reproducibility of the ELISA was in an acceptable range with a coefficient of variation under 1.5%. The ELISA was reliable in the range of 0.1 to 10 ng/mL. The polyclonal anti-rBPGM antibody did not show any cross-reaction with recombinant human B type phosphoglycerate mutase, which is highly homologous to rBPGM. The ELISA was found to be practical for the determination of BPGM protein content in human erythrocytes. The mean BPGM protein content was 56.3+/-9.7 microg/mL in whole blood (mean+/-SD, n = 50). The ELISA can be used to examine various hematologic disorders with abnormal red cell size and cell counts, and to detect BPGM enzymopathy in human erythrocytes.
Adult↗
A logic-based dynamical theory for a genesis of biological threshold.
To help the study of constructing a formal neuron by computer, we propose a logic-based dynamical theory for a genesis of the biological threshold which specific proteins, such as ion channel proteins, or their networks can produce. By viewing such a protein or a protein network as a computational machine, the statements concerning the states of reaction chains which eventually activate or inactivate the protein are treated. By introducing dynamical systems, associated with an inference process based on statements with continuous truth values, we investigated invariant characters of such dynamics and we obtained a sigmoidal function for an invariant distribution function of the truth values. The domain of solutions of the functional equations regarded as representing the self-description of proteins or protein networks as a machine indicates the emergence of a threshold, namely the realization of dyadic value, 0 or 1, based on the continuous truth values. The results obtained may highlight the mechanism of neuronal threshold in a framework which differs from population dynamics. The derived dynamical systems may also provide a simple model of 'demon' rectifying the thermal fluctuations to drive unidirectional movements.
Animals↗
[Current problems in quality control (QC) in hematology].
Total quality control (TQC) is one of the most important issues in laboratory hematology. As for complete blood count (CBC), almost all blood cell counters possesses quality control (QC) programs for its determination, and values of high quality can be provided by the counters. Reticulocytes can be determined precisely as well. However, mean cell volume (MCV) were slightly different among blood cell counters when semifixed QC materials were used for its surveillance. No QC materials to evaluate white cell differential functions have not be found for the common use now. Thus, urgent matter would be the development of appropriate QC materials to be used for overall blood cell counters. In coagulation testings, trials for common use of international normalized ratio (INR) for the prothrombin time (PT) test has been continued yearly due to the varieties of biophysical characteristics among PT reagents. Similar variation are also present among activated partial thromboplastin time (APTT) reagents. Some improvement would be required.
Blood Cell Count↗
[Effective utilization of simple testing instruments equipped with microanalytic technology].
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Clinical Chemistry Tests↗
Polymorphonuclear neutrophils in saliva and blood: a comparative study of morphology, function and phenotype.
The morphology, phagocytic activity, production of hydrogen peroxide (H2O2) and expression of lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18), complement receptor type 3 (CR3; CD11b/CD18), p150,95 (CD11c/CD18), and platelet/endothelial cell adhesion molecule-1 (PECAM-1; CD31) by oral polymorphonuclear neutrophils (PMN) are assessed and the results compared with those of blood PMN. There were no differences in the morphology and phagocytic activity between oral and blood PMN. H2O2 production was measured following stimulation with phorbol myristate acetate (PMA) and N-formyl-L-leucyl-L-phenylalanine (FMLP) as indicators of bactericidal activity. There was a significant difference in the H2O2 production by the two groups when stimulated with FMLP; the level of H2O2 production by oral PMN was significantly higher than that by blood PMN. However, there was no significant difference in H2O2 production between oral and blood PMN when stimulated with PMA. The percentage of CD11a- and CD11c-positive concentrated oral PMN was significantly lower than that seen in blood PMN, as was the percentage of CD31-positive cells. Higher H2O2 production by oral PMN following stimulation with FMLP may result in enhanced bactericidal activity. Low expression of CD31 may lead to the accumulation of PMN in the mouth by blocking their return to the bloodstream. These phenomena may be necessary for oral PMN to protect periodontal tissues from bacteria in the mouth.
Cell Adhesion Molecules↗
A new type of self-organization associated with chaotic dynamics in neural networks.
A new type of self-organized dynamics is presented, in relation with chaos in neural networks. One is chaotic itinerancy and the other is chaos-driven contraction dynamics. The former is addressed as a universal behavior in high-dimensional dynamical systems. In particular, it can be viewed as one possible form of memory dynamics in brain. The latter gives rise to singular-continuous nowhere-differentiable attractors. These dynamics can be related to each other in the context of dimensionality and of chaotic information processings. Possible roles of these complex dynamics in brain are also discussed.
Animals↗
Measurement of Fe2+ ion by coulometry method at incubation of Thiobacillus ferrooxidans.
Thiobacillus ferrooxidans is a chemoautotrophic bacterium that is capable of using Fe2+ oxidation by O2 as the sole source of energy for growth and CO2 fixation. The idea of the solar bacterial biomass farm by using of this bacterium is proposed. The incubation experiment of these bacteria was carried out, and the 9K culture medium as the standard medium for T. ferrooxidans was used. The measurement of Fe2+ in the growth stage was carried out as the first step of the experiments to clarify the possibility of this system. The items of measurement were Fe2+ ion density, pH of the medium, bacterium density and quantity of total organic carbon (TOC). The density of Fe2+ ion in the medium was measured by coulometry method. This method has the following advantage, high accuracy (<1%), easy operation, short measurement time (a few minutes) and small sample quantity (about 0.1 ml). The experimental results show that the Fe 2+ ion density is measured as same as the accuracy of pH measurement. At the final stage of the growth, the pH decreased due to the generation of the iron hydroxide (Fe(OH)3). The bacterium density and TOC slightly increased after that Fe2+ runs short. This result shows that the CO2 fixation speed is slower than Fe2+ oxidation speed. It is shown by the experiment that the growth limit of T. ferrooxidans is caused by the disappearance of the Fe2+ ion. It may be possible that the bacterium density increases by the continuous supply of Fe2+ ion.
Biomass↗
DNA extraction and amplification from Giemsa-stained blood smears.
DNA extraction was attempted from Giemsa-stained blood smears on glass slides that had been stored for several years. High molecular weight DNA bands were clearly visible after electrophoresis when DNA was extracted from specimens stored up to 2 years. Specimens more than 4 years old demonstrated a smeared pattern, suggesting degeneration of the DNA, but they could be rescued by PCR amplification using primers of HLA-DQA1 genes. The recovery could be pursued even in 11-year-old specimens.
Azure Stains↗
Inaccuracy and inefficiency of urinary sediment analysis.
When the efficiency of urine sediment analysis was studied: we found that: (1) In tests of repeatability, counts of red blood cells, white blood cells, and epithelial cells, but not of casts, showed high precision, even when they were conducted by relatively inexperienced technologists; (2) Correlation of counts of red blood cells, white blood cells, and epithelial cells was good for results obtained by an expert technologist and those obtained by relatively inexperienced technologists, but the correlation was low in the case of casts, perhaps because of their low incidence in urine sediment; (3) Hematuria and leukocyturia were often detected in specimens positive for occult blood, albumin, or both; (4) Most casts were hyaline, which are physiologic in nature. Pathologic casts such as granular casts, were found in specimens containing red and white blood cells; (5) Although urine sediment analysis was frequently requested by clinicians, most analyses showed negative results; (6) Microscopic sediment analysis requires considerable time, labor, and experience, and is more sensitive than tape tests. The results of our study indicate that there are advantages to the present method for analyzing urine sediment, but that its efficiency is doubtful.
Humans↗
Detection of microplatelet formation using an electroimpedance cell counter.
Changes in the platelet size distribution curves during the process of ADP-induced aggregation were observed using a particle distribution analyzer (PDA) based on electronic impedance. The maximum primary aggregation was seen at a final ADP concentration of 3 microM. After stimulation, the platelet distribution curve changed rapidly from a log normal curve to a curve skewed to the left. When the changes were evaluated at 1 min after the addition of ADP, the peak height was increased, and the median and total volume were decreased significantly. These findings indicate that microplatelets are formed by membrane stimulation and this formation can be monitored with the use of this automated instrument.
Adenosine Diphosphate↗
Flow cytometric detection of recombinant human granulocyte-colony stimulating factor binding to leukemic cells.
To examine binding of recombinant human granulocyte-colony stimulating factor to myeloid cells, the factor was labeled with fluorescein isothiocyanate, and incubated with blood specimens, which were then analyzed by flow cytometry. Neutrophils demonstrated an increased fluorescence, while lymphocytes were negative. These cell fractions were used as controls for cytometric binding assays of leukemic cells. Six patients with lymphocytic leukemia were negative in this assay. Ten of 15 patients with myelocytic leukemia were positive. All patients (n = 5) in myeloblastic crisis of chronic myelogenous leukemia were also positive. The flow cytometry results correlated well with the results of colony formation in response to granulocyte-colony stimulating factor. The results indicate that our method is useful in predicting the susceptibility of leukemic cells to recombinant growth factors.
Flow Cytometry↗
Reassessment for reference values for total nucleated cell count and myeloid/erythroid ratio of human bone marrow aspirate.
Total uncleated cell count and Myeloid/Erythroid (M/E) ratio of bone marrow aspirate in Japanese was determined in 1962 and no reassessment has been done until now. Therefore, the reassessment work has been performed using our laboratory data. Criterias for healthy individuals were settled and data obtained were applied first for probability plot analysis chart and data which gave asymmetry for the distribution curve were expelled. Then the data sets were processed with Hoffman's 3 time cutting method. As a result, reference values of the count and the ratio were obtained in total and by gender and age. The former values were lower than those reported by previous papers, but the latter values were similar to those reported previously. On the other hand, our manual counting method was compared with the electronic counting method, and good correlation was observed between the both. From the results we concluded that our reference values were acceptable and useful for routine use.
Bone Marrow Cells↗
[Standardization in flow cytometry].
Analysis of lymphocyte subsets and determination of leukemic cells by flow cytometry has now become an important clinical test. With recent technological developments, reliability was improved and the procedure became easier. To establish quality assurance procedures for flow cytometry, standardization is necessary. In the United States, the National Committee for Clinical Laboratory Standards (NCCLS) has been working actively and has published several tentative guidelines. However, in Japan, such work lags behind that in western countries. The results of quality control surveys show that differences in data exist among instruments and laboratories. To minimize such differences, nation-wide surveillance must be established. Also, efforts should be made to establish reference methods and reference materials that will lead to standardized results. We hope standardization will ensure precision and accuracy in flow cytometric results in clinical laboratories.
Antibodies, Monoclonal↗
[Standardization in hematology and the use of the data in clinical laboratories].
Recently developed automated blood cell counters provide accurate and precise data on complete blood count (CBC). The results of nationwide surveys obtained using them are usually better than those obtained by biochemical tests. The precision of CBC counting has been improved owing to the efforts of laboratories and manufacturers. As for hemoglobinometry, the cyanmethemoglobin and oxyhemoglobin methods used in blood cell counters gave different results for fresh and artificial blood in control surveys. Now instead of the oxyhemoglobin method, the sodium lauryl sulfate-hemoglobin method is used to minimize the variation among different counters. The biggest problem at present is the lack of control blood which can be used for any blood cell counter. Another problem is the laboratory report sent to clinicians, which gives only numerical data. However, other comments and normal reference values should also be presented. Sufficient care may not be taken for normal reference values; they vary considerably among different laboratories. Also, most laboratories and manufacturers now use various conventional units for CBC, but the SI unit should be used for comparison of the data. In conclusion, nationwide unification of CBC values should be established in the near future with cooperative efforts by laboratories and manufacturers.
Blood Cell Count↗
Measurement of the zeta potential of human platelets by the use of laser-light scattering.
An instrument was developed to detect the shift in scattering of laser light that occurs when particles in suspension move in a chamber with an electrical load. The instrument measures the zeta potential of particles. We applied the instrument to study human blood cells. Platelet-rich plasma was used because of the stability of the suspension, without the sedimentation or autoaggregation that is often seen with red or white blood cells. The reproducibility of the measurements was satisfactory when there were enough platelets in the suspension. Platelets from healthy controls (n = 136) had a potential of -14.20 +/- 1.64 mV at the detection angle of 17.1 degrees. Platelets from patients with essential thrombocytosis (n = 16) or polycythemia vera (n = 8) had higher potentials than the healthy controls.
Blood Platelets↗
Automatic measurement of hemoglobin F in blood obtained from patients with hemoglobin E/E and beta-thalassemia/hemoglobin/E.
This paper presents an automated determination of hemoglobin (Hb) F in Hb E/E disease using Hi-Auto A1c. Blood specimens collected in Bangkok were frozen, and sent to Japan by air mail for the determination. The automatically determined values showed a high correlation with the values obtained by the classical alkali denaturation method. Hb E/E cases showed 4.24 +/- 1.75% of Hb F. On the other hand, Hb, Hct, MCV and MCH in the disease samples were lower than in the controls, but higher than those of beta-thalassemia/HbE disease. From the results it was concluded that Hb E/E could be differentiated from beta-thalassemia/HbE by combination of Hb F value and MCH or Hb in CBC.
Adult↗