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Biomedical subjects

T Takubo

Publications and source records attributed to T Takubo.

At least 73 records · Page 4Linked to original sources

An effective method for recovering CD34 positive progenitor cells from peripheral blood stem cell apheresis products cryopreserved with simplified method.

The use of small volume collection chamber (SVCC) during peripheral blood stem cell (PBSC) apheresis, combined with simplified cryopreservation without rate-controlled freezing, have successfully been applied to clinical PBSC transplantation following myeloablative chemotherapies. However, the method to effectively select CD34+ progenitor cells from frozen apheresis products obtained with these simplified methods has not been reported. For this goal, after washing the thawed apheresis products with medium containing Dnase I, two different approaches to purify CD34+ progenitor cells from washed WBCs were compared. In method I, CD34+ cells were purified on the same day using immunomagnetic method. In method II, the cells after wash were cultured for overnight in RPMI-1640/10% FCS containing SCF and IL-3, followed by enrichment of CD34+ cells as in method I on the next day. In both methods, CD34+ cells were recovered with high viability. However, subsequent liquid culture revealed that the cells obtained from method II have clearly higher growth potential compared with the cells from method I. In addition, these CD34+ cells from method II well-tolerated to further refreeze and thaw. Thus, allowing to "rest" overnight after thaw may be critical for processing of the simply cryopreserved apheresis products as in method II.

Adult↗

Country reports from Japan: external quality assessment scheme in Japan.

Several external quality assessment schemes (EQAS) have been conducted in Japan. Results obtained from nation-scale EQAS reveal the current quality of laboratory testing in each laboratory. The largest nation-scale EQAS in Japan is that conducted by the Japan Medical Association. The numbers of participants and of items evaluated have increased in EQAS by JMA over its history of 32 years. Improvement in inter-laboratory differences has been observed for most items in EQAS in recent decades. In 1998, about 2,500 laboratories from throughout the country participated in this surveillance, and 47 items were evaluated. The coefficient of variations for the group of all participants was less than 5% for about one third of all test items. On the other hand, very high variations over 20% were observed for 6 items. Also, inter-method differences exist for many items, which may be or may not be related to matrix effects. Retrospective evaluation of all EQAS data suggests that there is still room for improvement in inter-laboratory differences.

Bacteriological Techniques↗

Incidence of abnormalities in laboratory tests found in surveillance of adults over 40 years of age.

National health check-up systems have been used for 5 years in Japan for adults who are over 40 years of age. As part of a national project, Osaka prefecture is also conducting a program for health check-up testing and cancer screening for this age group. This surveillance revealed that incidence of obesity, hypertension, cholesterolemia, albuminuria, or abnormal ECG was high. Analysis of surveillance results should contribute to understanding the present status and recent trends in diseases in the aged. With continuation of this surveillance for a number of years, trends in life-style related diseases in Japan should be detectable.

Adult↗

Quality control in a manual and an automated leukocyte differential count.

Quality control (QC) has been introduced in laboratories, and QC surveys in leukocyte differential count to enhance quality have been performed by College of American Pathologists, Japanese Association of Medical Technologists, Osaka Medical Association and manufacturers. The results of QC survey in a manual leukocyte differential count indicated problems on the differentiation of segmented neutrophils and band neutrophils and the detection of pathological blood cells on blood smear. While the results of QC survey in an automated leukocyte differential count performed by same manufacturer with an automated blood cell counter were satisfactory, however, there was a difference in leukocyte differential cell counts among laboratories with other manufacturer's instruments because the synthetic blood material used in QC is an exclusive item for an instrument. It is necessary to further reeducate the medical technologists in order to improve morphological performance, and to standardize the synthetic blood material for compatibility with various automated blood cell counters.

Automation↗

External quality control results for hormones, tumor markers and CRP testing.

Most hormones, tumor markers, C-reactive protein, and rheumatoid factor (RF) are measured immunologically. Immunological methods based on the antigen-antibody reaction have certain specific problems, including their principle of determination, character of antibodies used, reaction conditions, and others. Free thyroxine (FT4) and thyroid stimulating hormone (TSH), as well as alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), prostate antigen, carcinoantigen 19-9 (CA 19-9), and CA 125 are very commonly measured in the routine medical laboratory. Authentic materials can be obtained for hormones and CRP, and efforts to improve quality control and standardization have been made for years. Results of surveillance for FT4, TSH, and AFP were not poor, but inter-laboratory differences for CEA, CA 19-9, and RF were not insignificant.

Biomarkers, Tumor↗

Quality control in urinalysis.

Quality control (QC) has been introduced in laboratories, and QC surveys in urinalysis have been performed by College of American Pathologist, by Japanese Association of Medical Technologists, by Osaka Medical Association and by manufacturers. QC survey in urinalysis for synthetic urine by the reagent strip and instrument made in same manufacturer, and by an automated urine cell analyser provided satisfactory results among laboratories. QC survey in urinalysis for synthetic urine by the reagent strips and instruments made by various manufacturers indicated differences in the determination values among manufacturers, and between manual and automated methods because the reagent strips and instruments have different characteristics, respectively. QC photo survey in urinalysis on the microscopic photos of urine sediment constituents indicated differences in the identification of cells among laboratories. From the results, it is necessary to standardize a reagent strip method, manual and automated methods, and synthetic urine.

Automation↗

Localization of myosin, actin, alpha-actinin, tropomyosin and vinculin in surface-activated, spreading human platelets.

We observed the localization of the contractile proteins myosin, filamentous actin, alpha-actinin, tropomyosin, and vinculin in surface-activated, spreading human platelets using a single fluorescence staining procedure and conventional fluorescence microscopy. Myosin was distributed in a speckled pattern that extended radially from the granulomere. F-actin demonstrated cable-networks. Tropomyosin and alpha-actinin occurred in a punctuate distribution, and vinculin was localized at adhesion sites. Although myosin, F-actin, alpha-actinin, tropomyosin, and vinculin were not studied in resting platelets, our data support the idea that these contractile proteins are reorganized and reassembled in activated platelets during platelet function.

Actinin↗

Peripheral blood stem cell transplantation for non-Hodgkin's lymphoma in complete remission.

High-dose chemotherapy with PBSCT as consolidation therapy was administered to 15 patients with non-Hodgkin's lymphoma admitted to our department between November 1991 and March 1998. The average number of CD34-positive cells harvested was 3.1 x 10(6)/kg and the median time to a granulocyte count above 500/microliter was 12.8 days. Of the patients, one died from cytomegalovirus pneumomonia after WBC count recovered, 2 relapsed, and 12 patients are still alive and disease-free. The average disease-free survival period was 50.0 months. Stem cell harvesting is relatively simple and hematopoietic recovery is mory rapid with PBSCT than with ABMT. Our findings suggest that PBSCT can be used for consolidation therapy in NHL patients who have achieved complete remission with standard chemotherapy.

Adolescent↗

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↗