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Biomedical subjects

I Tsuda

Publications and source records attributed to I Tsuda.

At least 19 recordsLinked to original sources

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

Size distribution curves of blood cells in thalassemias and hemoglobin H diseases.

Thalassemias and hemoglobinopathies in Thailand have been examined with a blood cell counter based on electroimpedance principle and obtained size distribution curves of red cells and platelets. Among various disorders, beta-thalassemia/hemoglobin E and homozygous hemoglobin Constant Spring showed severe anemia. Their red cell size distribution curve shifted to the left and overlapped with the platelet size distribution curve. Red cell distribution width expressed by coefficient of variation and the degree of the overlapping were stronger in beta-thalassemia/HbE than HbH. Heterozygous beta-thalassemia showed a narrow red cell distribution curve width with small standard deviation and low England's value. Although the overlapping of size distribution curves cause inaccurate red cell count and platelet count, careful observation of the size distribution curves was proved to have high diagnostic value.

Blood Platelets

Reticulocytes in human preserved blood as control material for automated reticulocyte counters.

The authors studied the changes in the percentage of reticulocytes in blood preserved for three weeks. The samples were obtained from healthy adults and stored at 4 degrees C. An automated reticulocyte counter was used for measurements. After one week, the relative percentage of reticulocytes had decreased to around 80% of the initial value, and it reached 60% after three weeks of storage. When the y-axis of plots of the change against time was converted to a log scale, negative correlation was strong (r = -0.9972), and the curve was useful for estimation of the reticulocyte count in preserved blood. Stored blood might be used as a control material for reticulocyte counting by automated methods.

Adult

An automated optoelectronic reticulocyte counter.

Microscopic reticulocyte counting is time consuming and imprecise. A new reticulocyte counter has been developed, and the authors evaluated its utility for laboratory use. The counter, R-1000 of Sysmex-TOA Medical Electronics Company, Kobe, Japan, is based on the principles of flow cytometry. Reticulocytes are detected as fluorescent cells stained with a basic dye, auramine O, under argon-laser light. The automated count had high correlation to the manual count (r = 0.941). Linearity and reproducibility were both high. About 60 specimens were tested in one hour. Not only the reticulocyte percentage and count but also the maturity of reticulocytes was found from the intensity of the fluorescence, whether high, moderate, or slight. Normal reference values were 0.007 +/- 0.0055 (0.70 +/- 0.55%) for the reticulocytes, (4.63 +/- 1.09) X 10(9)/L for the reticulocyte count, 2.3 +/- 1.9% for highly fluorescent cells, 18.7 +/- 5.1% for moderately fluorescent cells, and 78.8 +/- 6.6% for cells with slight fluorescence. In patients with suppressed bone marrow function, such as is caused by chemotherapy, the reticulocyte fraction and count were low, and cells with slight fluorescence increased. In patients in whom bone marrow function was stimulated, such as with hemolytic anemia, the reticulocyte percentage, reticulocyte count, and highly fluorescent cells were high. Patients with chronic renal failure being treated by hemodialysis had a similar reticulocyte pattern to that in hemolytic anemia except that the reticulocyte count was decreased. Results for elderly patients were not different from those of healthy young controls. Some patients with a normal reticulocyte count and percentage had numerous highly fluorescent cells, perhaps because of hemolytic anemia not yet identified. Automated reticulocyte counting provides reliable data, so such measurement should be useful for analysis of the kinetics of red blood cells and for the study of the pathogenesis of anemia.

Adult

Trial of ABO and Rh blood typing with an automated blood cell counter.

We designed a method for ABO and Rh typing using a blood cell counter. Blood specimens were diluted with physiological saline, mixed with an antiserum, and incubated. The cell count and size distribution curves were obtained by an automated blood cell counter. The samples with agglutination had a decreased cell count and a characteristic size distribution curve with a population of doublet cells on the right side. The results were correlated with the degree of agglutination observed by the manual method, used as a reference. It should be possible to adapt blood cell counters for use in blood typing.

ABO Blood-Group System

Inaccuracy and imprecision of reticulocyte counting.

Reticulocyte count is an essential clinical test to detect erythropoietic activity. The test has been done manually with the use of one of three dyes: new methylene blue, brilliant cresyl blue, and azure B. When reticulocyte counts with the different dyes were compared, correlation was good. When reticulocyte counts obtained by use of the dyes were compared with the count obtained by an automated counter, correlation was high. In a sampling study, the specimen prepared by a spinner method gave smaller variations than a wedge method. Intertechnologist bias was higher than inter-specimen or intra-specimen bias: The variation of the reticulocyte count was examined with from 200 to 2,000 cells; with 1,000 cells or more, variation was low. From these results, we concluded that more than 1,000 erythrocytes on one slide prepared by the spinner method should be analyzed. Clear standards for cell identification are also needed.

Blood Cell Count

Analysis of hematological data of thalassemia cases in Thailand.

We analyzed the complete blood count (CBC) obtained from patients with various types of thalassemia in Thailand. As a control, samples from healthy donors and patients with iron deficiency anemia were also collected. Most cases of thalassemia could be differentiated from iron deficiency anemia by using a discrimination function equation. This screening method is applicable for mass-screening of patients with latent thalassemia in Thailand.

Erythrocyte Count

An automated reticulocyte counting method: preliminary observations.

We evaluated the counting of reticulocytes in the peripheral blood with a newly developed flow cytometer type of automated counter that performs a single test within 60 s. The volume of sample needed is 100 microliters and the cells are stained with auramine-O in the counter. The mean within-run reproducibility was 4.66% (CV, n = 50), and dilution of blood gave highly linear results with an r value of 0.996. Correlation was good between manual reticulocyte counts and those performed with the counter (r = 0.893). Samples with large numbers of leucocytes, erythrocytes, or platelets did not interfere with the automated reticulocyte counting, and provided accurate and precise data.

Autoanalysis

Electric sizing of platelets stimulated by adenosine diphosphate (ADP).

The platelet-rich plasma stimulated by a low dose of adenosine diphosphate (ADP) was examined by an electric counter, oscilloscopy, an electron microscopy and packed cell volume analysis. The size distribution curve obtained by the electric counter shifted to the left, and the cell volume was decreased. On the oscillograph, those small cells were seen as pulse signals with a regular shape. However, microscopic observation did not find as many microplatelets or fragments. Also, packed cell volume analysis showed no difference between ADP-stimulated and unstimulated samples. On the electronmicrograph, a slight aggregation was seen, with many of the ADP-stimulated platelets exhibiting spiny forms. The shape factors influences strongly the electric sizing, thus, the deformation of platelets may be the reason for the disagreement between the results by electric observation and those by other methods. Our results indicated that the small platelets observed on the size distribution curve do not always reflect the actual small cells.

Adenosine Diphosphate

Leukemic blasts detected by the Technicon H-1 blood cell counter.

The authors studied the usefulness of the flagging system of the Technicon H-1 Cell Counter using samples from 45 patients with acute leukemia, 24 with acute lymphoblastic leukemia (ALL), and 21 with acute nonlymphoblastic leukemia (ANLL). Results by the automated method were compared with those by the eye-count method. Four out of 24 specimens were falsely negative; no false positive results were found. For ANLL specimens, there were no false positives or negatives. When 1% or more of the cells were blasts by the eye-count method, the automated system almost always flagged the sample. The results suggested that the flagging system programmed in the H-1 is useful for the diagnosis and follow-up of leukemia.

Blastomeres