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Biomedical subjects

Takayuki Takubo

Publications and source records attributed to Takayuki Takubo.

At least 19 recordsLinked to original sources

Detection of various platelet-associated immunoglobulins by flow cytometry in idiopathic thrombocytopenic purpura.

BACKGROUND: In 1975, Dixson reported that anti-platelet IgG on platelets from patients with idiopathic thrombocytopenic purpura (ITP) is greater than in normal people, by determining anti-platelet antibodies directly on the platelet surface with a quantitative complement lysis-inhibition-assay. Since then, platelet-associated IgG (PAIgG) has been thought of as evidence of ITP. Although platelets from ITP patients show significantly higher PAIgG values than from normal control individuals, PAIgG is not specific for autoantibody because it increases in other than immune ITP patients. METHODS: We analyzed positive platelet percentage with various platelet-associated immunoglobulins: IgG, IgM, IgA, and total immunoglobulins, in the blood from 17 normal donors and 23 ITP patients. RESULTS: The specificity for ITP disease was better in flow cytometry than in ELISA, because, other than ITP, only aplastic anemia was positive in flow cytometry; however, various disorders (aplastic anemia, chronic lymphocytic leukemia, acute myeloid leukemia, and myelodysplastic syndrome) showed positive in ELISA. Flow cytometry methods had the same sensitivity for ITP disease as ELISA. However, it is supposed that there was no nonimmune ITP in this study because the PAIgG negative patients (n = 1) showed positive results in flow cytometry. CONCLUSION: Flow cytometry method was effective for ITP screening, especially for specificity.

Adult↗

Evaluation of noninvasive hemoglobin monitoring for hematological disorders.

The Astrim is a noninvasive hemoglobin monitoring device that uses near-infrared radiation. We studied the usefulness of this device for patients with hematological disorders. We measured hemoglobin levels 309 times in 140 subjects, using an automated hematology analyzer (K-4500) and the Astrim. The coefficient of correlation between Hb levels with the K-4500 (K-Hb) and the Astrim (A-Hb) was r = 0.591 (P < 0.001). The coefficient of correlation between A-Hb and K-Hb from 174 specimens with anemia and 135 without anemia was r = 0.531 (P < 0.001) and r = 0.345 (P < 0.001), respectively. In the 309 specimens, the Astrim showed a diagnostic sensitivity and specificity for anemia of 78.3% and 69.0%, respectively. While the r-values, sensitivity, and specificity were passable, they appeared to be insufficient for an accurate evaluation. We believe this was caused by the measurement conditions (i.e., the finger selected for measurement, the finger position in relation to the CCD camera, and the finger temperature). On the other hand, the precision test results were good. Therefore, if careful attention is paid to measurement conditions, the noninvasive Astrim is clinically useful for continuous hemoglobin monitoring of hematological disorders and patients with hemorrhagic diseases, such as perioperative hemorrhage and gastrointestinal bleeding. However, it is difficult to determine Hb concentrations in one measurement using the Astrim because the r-values, sensitivity, and specificity are insufficient for accurate evaluations of in-patients, particularly patients with advanced diseases. Therefore, we believe it is necessary to improve the system so that A-Hb can be accurately determined in one measurement.

Adolescent↗

[Effect of glucocorticoids on bone mineral density in patients with idiopathic thrombocytopenic purpura].

To elucidate the effects of glucocorticoid on bone mineral density in idiopathic thrombocytopenic purpura (ITP) patients, we retrospectively evaluated the relationship between bone mineral density (BMD) and the total dose of glucocorticoid or the mean daily dose given. We found decreased BMD in 66.7% of the patients with ITP to whom glucocorticoid was given, though rather normal bone BMD was observed in 28.6% of ITP patients treated without steroids. The mean level of BMD was markedly decreased in steroid-treated patients compared with steroid non-treated patients (p < 0.01). The relationship between BMD and the total dose of glucocorticoid (p = 0.023) or the mean daily dose revealed a negative correlation (p = 0.022). This study showed that glucocorticoid-induced osteoporosis was observed in patients with ITP, similar to other diseases already reported. When we think of this disease, many cases tend to be followed for a long time, and as the majority of ITP patients is female, we should pay particular attention in the prevention of glucocorticoid-induced osteoporosis.

Adult↗

A case of a long-time survivor with chronic active Epstein-Barr virus infection.

Epstein-Barr virus (EBV) is associated with hypersensitivity to mosquito bites (HMB) and fatal EBV-associated hemophagocytic syndrome (HPS). The prognosis of patients with chronic active EBV infection (CAEBV) is very poor. We report a rare case of an adult woman patient with a 28-yr history of HMB, who developed EBV-HPS. EBV genome was detected in the serum and peripheral blood lymphocytes. Clonal proliferation of EBV was demonstrated by Southern blot analysis using an EBV genome terminal-repeat probe. This is a very rare case of a long-term survivor with CAEBV. The patient was initially treated with immunochemotherapy and achieved complete remission. However, the patient immediately relapsed and underwent allogeneic bone marrow transplantation (BMT) from her HLA-matched brother. Peripheral blood cell recovered well, and EBV genome disappeared from the peripheral blood. Allogeneic BMT may be effective in eradicating EBV-HPS. Unfortunately, the patient died of graft vs. host disease on the 92nd day after BMT.

Adult↗

Automatic analysis of normal bone marrow blood cells using the XE-2100 automated hematology analyzer.

The bone marrow aspiration test conventionally has been performed by visual methods, using a light microscope, because automatic blood cell analyzers cannot adequately capture erythroblasts and immature granulocytes (IGs) (Tatsumi et al.: Osaka City Med J 1988;34:135-146; Tatsumi et al.: Am J Clin Pathol 1986;86:50-54). With the development of the XE-2100 automatic blood cell analyzer (Sysmex Corporation, Kobe, Japan) in 1999, the classification of erythroblasts and IGs by means of flow cytometry (Zini et al.: Infus Ther Transfus Med 2001;28:277-279; Briggs et al.: Sysmex J Int 1999;9:113-119) became possible. In the present study we classified cells in 65 bone marrow aspiration specimens by the microscopic method and with the XE2100, and compared the results. A good correlation was found in the nucleated red blood cell (NRBC), white blood cell (WBC), and total nucleated cell (TNC) counts; the myeloid/erythroid ratio (M/E ratio); neutrophils, lymphocytes, eosinophils, and IGs in the immature myeloid information (IMI) channel; and the total cell count. These items can all be analyzed in about 54 sec with the XE2100, which is faster than with the microscopic method. Therefore, analysis of bone marrow aspiration specimens with this apparatus appears to be very useful for clinical screening as well as laboratory testing.

Biopsy, Needle↗

Reduced-intensity stem cell transplantation in two cases of metastatic renal cell carcinoma.

Allogeneic hematopoietic stem-cell transplantation can induce curative graft-versus-leukemia reactions in patients with hematological malignancies. There is also evidence of such an effect in patients with solid tumors. We report two patients with metastatic renal cell carcinoma who underwent RIST. In both patients, disease progression was observed 6 months after transplantation. However, one patient had transient symptoms of tumor progression after the occurrence of acute graft-versus-host disease, consistent with graft-versus-tumor effects.

Bone Neoplasms↗

[Analysis of factors for poor mobilization after administration of recombinant human granulocyte stimulating factor (rHuG-CSF) in healthy donors].

Allogeneic hematopoietic stem cell transplantation is an effective treatment for hematological malignancies. Peripheral blood stem cells (PBSCs) are increasingly used as an alternative to bone marrow for allogeneic transplantation. However, predictive factors for the response to recombinant human granulocyte stimulating factor (rHuG-CSF) in healthy donors have not been extensively studied. We analyzed the side effects, laboratory test results after administration of rHuG-CSF and the factors influencing mobilization of peripheral blood stem cells in 30 healthy donors. Bone pain, fever and headache were observed with high frequency after administration of rHuG-CSF. WBCs and reticulocytes increased, and RBCs and platelets decreased significantly after administration rHuG-CSF. Biochemical examination revealed significant elevations of LDH, CRP, ALP and UA. Univariate analysis showed the age of donors (< 50 vs. > 50, p = 0.041) and the lymphocyte counts before administration of rHuG-CSF (p = 0.032) to be correlated with the number of CD34 positive cells. From a multivariate analysis, the tendency for good mobilization with a twice daily dose of rHuG-CSF (p = 0.065) was observed. The rHuG-CSF schedule may be the most important factor affecting peripheral blood stem cell mobilization and collection in healthy donors.

Adolescent↗

[Successful treatment with foscarnet for disseminated varicella-zoster infection after reduced intensity stem cell transplantation in a case of relapsed refractory central nervous system lymphoma].

Visceral disseminated varicella-zoster virus (VZV) infection occurred with acute graft-versus-host disease in a 33-year-old Japanese male with non-Hodgkin lymphoma who had undergone allogeneic stem cell transplantation from an HLA-identical sibling after reduced intensity conditioning chemotherapy. Although ganciclovir and acyclovir treatment was effective temporarily, the number of VZV-DNA copies in the blood remained at a high level, and the hepatitis was prolonged. The patient was treated with foscarnet, which led to improvement of the VZV viremia and the hepatic dysfunction. Foscarnet therapy should be considered for acyclovir-resistant VZV infection in the setting of allogeneic hematopoietic stem cell transplantation.

Acyclovir↗

[An XXX female with essential thrombocythemia].

We describe an XXX female patient accompanied with essential thrombocythemia. To our knowledge this is the first case ever to have been reported. The patient was asymptomatic, but her platelet count had increased to 111.2 x 10(4)/microliter, and she was diagnosed as having essential thrombocythemia based on the diagnostic criteria of the Polycythemia Vera Study Group. At the same time, chromosome analysis of bone marrow cells revealed that she was an XXX female. The patient remained asymptomatic throughout the course of treatment.

Chromosomes, Human, X↗

Simple high-density lipoprotein cholesterol assay based on dry chemistry.

BACKGROUND: Currently, high-density lipoprotein cholesterol (HDL-C), a factor which prevents progression of arteriosclerosis, is measured using laboratory-based chemistry analyzers without a pretreatment step. Because HDL-C is measured with a pretreatment step in many point-of-care testing systems, a direct assay is needed. METHODS: A dry-chemistry-based assay using surfactants has recently been developed in parallel with the development of a dedicated reagent. A simple analyzer that accepts whole blood samples was also developed. RESULTS: The assay demonstrated excellent precision, dilution linearity and intermethod comparison. In an interference test, assay values tended to be lower in the presence of high concentrations of hemoglobin, conjugated or unconjugated bilirubin. Neither ascorbic acid up to 20 mg/dl, nor formazin turbidity up to 2100, had an effect on the assay. CONCLUSIONS: This dry-chemistry assay using only surfactants for specificity in the direct HDL-C method was judged useful for point-of-care instrumentation in terms of equipment compactness, operational simplicity and rapid responsiveness.

Blood Chemical Analysis↗

Differential effects of fibroblast growth factor-4, epidermal growth factor and transforming growth factor-beta1 on functional development of stromal layers in acute myeloid leukemia.

The hematopoietic supporting abilities are known to be impaired in marrow stromal layers developed from patients with acute myeloid leukemia (AML). In this study, fibroblast growth factor-4 (FGF-4), epidermal growth factor (EGF) or transforming growth factor-beta1 (TGF-beta1) were studied to see whether these growth factors can modify the functional development of leukemic stromal layers. Adherent stromal layers from 13 patients with AML and from six non-leukemic controls were established with 3ng/ml of FGF-4, EGF or TGF-beta1. Established stromal layers were washed three times and irradiated, followed by recharge of allogenic peripheral CD34 positive cells as an indicator of supportive function. Progenitor-outputs into supernatant were evaluated at biweekly interval with colony-forming assay until 6 weeks. The results showed that both leukemic and non-leukemic stromal cells established with FGF-4, but not with EGF, showed significantly higher progenitor cell-outputs compared with control stromal cells. By contrast, stromal cells developed with TGF-beta1 showed significantly lower progenitor cell-outputs compared with control. These differences were significant at later than 4 weeks after the recharge of indicator cells, suggesting that the stromal layer developed with EGF or TGF-beta1 preferentially affected the primitive progenitors rather than committed ones. These results indicate that FGF-4 and TGF-beta1 differentially affect the functional development of leukemic as well as of normal stromal layers.

Acute Disease↗

Combination assay of urinary free L-fucose and trypsin inhibitor may be useful indicators of disease activity in patients with hematologic malignancies.

L-Fucose is a monosaccharide located at the non-reducing ends of sugar chains of glycoconjugates. Urinary L-fucose (U-FC) is excreted as free L-fucose, and clinically useful as a tumor marker of digestive organ cancers. We evaluated the clinical usefulness of U-FC levels in patients with various hematologic malignancies because U-FC for hematologic malignancies has only rarely been described. The mean U-FC levels in the acute nonlymphocytic leukemia (ANLL), myelodysplastic syndromes (MDS) and myeloproliferative disorders (MPD) groups were significantly higher than in the control group (P < 0.05). Recently, we reported that urinary trypsin inhibitor (UTI) levels in patients with ANLL, MDS, non-Hodgkin's lymphoma and multiple myeloma were significantly elevated, compared with those in healthy adult volunteers. Noninvasive combination assays of UTI and U-FC may have a higher accuracy in diagnosis of ANLL and MDS than those of UTI or U-FC alone. UTI and U-FC combination assays, noninvasive for patients, could be expanded as useful indicators in hematologic malignancies.

Biomarkers↗

[A case of amelanotic melanoma of the nasal cavity].

A 68-year-old male visited Hospital A for treatment of epistaxis, his chief complaint. He was told that he had an easily-bleeding tumor in the nasal cavity. Based of biopsy, a diagnosis of amelanotic melanoma was made. Operation was performed for removal of the tumor. About 8 months after discharge, he visited Hospital B with complaints of lumbar pain and epistaxis. After biopsy at Hospital B, malignant lymphoma (diffuse large cell) was diagnosed, and the patient was referred to our hospital. On bone marrow puncture and biopsy, tumor cell infiltration was observed. Flow cytometric surface marker analysis revealed that these tumor cells were negative for CD45. Results of HE staining of the nasal cavity tumor were insufficient for diagnosis, and staining by immunohistochemistry was necessary to confirm the diagnosis. On immunohistochemical staining of the nasal cavity tumor tissue and bone marrow biopsy tissue, LCA, L26 and UCHL-1 were negative, and S-100 and HMB-45 positive. Recurrence of amelanotic melanoma accompanied by bone marrow infiltration was therefore diagnosed. The incidence of amelanotic melanoma with primary lesions in the nasal cavity is low. However, in making the diagnosis of a nasal cavity lesion, the possibility of such a melanoma should be kept in mind. In many cases, it is difficult to diagnose amelanotic melanoma with HE staining alone, and immunohistochemistry must be used.

Aged↗

[Reticulated platelet and its clinical significance].

Reticulated platelets (RP) retain some residual mRNA in their cytoplasm and are thought to be the most recently produced platelets in circulation. They can be visualized on a blood film with new methylene blue staining. RP is a flow cytometric assay utilizing a fluorescent dye, either thiazole orange or auramine O. There is a difference in reference values for RP between thiazole orange and auramine O. From the analytic results of RP in patients with thrombocytopenic disorders, RP measurement is considered useful for estimating thrombopoiesis in bone marrow and for differential diagnosis and elucidating the pathophysiology of thrombocytopenic disorders. In the future, test for RP might have a possibility of routine examination because RP can be rapidly and simply measured using whole blood stained with auramine O using an automated reticulocyte counter modified to determine RP. Furthermore, it is anticipated that standardization and morphological definition for RP are established and accuracy for RP is improved.

Blood Platelets↗

Problems on national and locale external quality assurance schema (NEQAS and LEQAS); based on Japanese experience.

The quality and stability of surveillance materials is a critical issue to perform good external quality assessment schema. In the case of nationwide surveillance, more stable materials are required than those in the case of locale external quality control schema. For locale surveillance, fresh blood, plasma or serum materials are prepared for hematology and chemistry tests. On the other hand, dried materials are used for the chemistry test and semi-fixed blood is used for hematology tests in the national surveillance. However to evaluate automated white cell differential in hematology, there has been no good materials. We therefore prepared a mixture of EDTA and ACD solution for our locale surveillance. It gave us excellent results on the differential when we used it within 3 days after blood collection.

Chemistry, Clinical↗