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Biomedical subjects

E Tatsumi

Publications and source records attributed to E Tatsumi.

At least 19 recordsLinked to original sources

Amorphism and physicochemical stability of spray-dried frusemide.

The physicochemical properties of amorphous forms of frusemide, prepared by spray-drying at 50 or 150 degrees C, and their hygroscopic stability at temperatures of 25 and 40 degrees C, and at 0 and 75% relative humidity were investigated. The glass transition temperature of the amorphous form A was 44.2 degrees C as measured by differential scanning calorimetry, while that of the amorphous form B was 54.4 degrees C. The activation energies for glass transition and crystallization processes were calculated from the differential scanning calorimetry thermograms of amorphous forms A and B, respectively. Stability determined by X-ray diffraction at 0% relative humidity, 25 and 40 degrees C suggested that form B was more stable than form A. However, the stability of form A at 75% relative humidity and 25 and 40 degrees C was similar to that of form B.

Drug Stability

Detection of Epstein-Barr virus genome in natural-killer-like cell line, YT.

YT cells, originally reported as a natural-killer-like (NK-like) lymphoid cell line, were investigated for Epstein-Barr virus (EBV) genome, gene rearrangement for T-cell receptor (TCR), phenotype and function. The YT cells of the original batch (YT-0) and two subclones (YT2C2 and YTC3) expressed EBV-associated nuclear antigen, and the BamHI-digested DNA showed the 3.4 kb hybridizing band with the BamHIW probe of EBV DNA in Southern blot analysis. When tested with latent-infection membrane protein probe, an identical hybridizing band was shared, indicating that all three sources of YT cells were of monoclonal derivation in terms of the terminal repeat junctional structure of EBV DNA, and that the original YT cells had been infected with EBV before the isolation of the two subclones. The cell-surface antigen analysis revealed the expression of CD7, CD28, CD30, CD45R0, TLiSA and S6F1 antigen besides the originally recorded CD25, CD56 and HLA-DR antigen. Gene rearrangement analysis showed the germ-line genotype, including TCR gamma and delta as well as beta chain. The Northern blot study using the CD3 epsilon and CD3 delta chain gene probes revealed CD3 epsilon, but not CD3 delta RNA. The YT-0 cells exhibited NK and antibody-dependent cellular cytotoxicity activity, but the YT2C2 and YTC3 cells did not. It was not resolved whether the fresh neoplastic NK-like cells of the YT-cell donor carry EBV genome, but YT cells, the first lymphoid cell line found to have EBV genome and non-B lymphoid properties, are valuable for investigating the relationship between EBV and human non-B lymphoid hematopoietic cells.

Antigens, Surface

Epstein-Barr virus (EBV) and human hematopoietic cell lines: a review.

Two facts need to be pointed out to help explain why the history of Epstein-Barr virus (EBV) research has been inseparable from that of the studies with human hematopoietic cell lines of neoplastic and non-neoplastic origin. One is that Burkitt lymphoma (BL) cell lines, EBV-positive or-negative, can be established in culture quite easily. Thus, the BL cell lines which Epstein established were indeed some of the first hematopoietic as well as virus-carrying cell lines of human neoplastic origin. The other is that EBV-positive B-cell lymphoblastoid cell lines (B-LCL) of normal origin can be grown from samples of sero-positive individuals. B-LCL were often mistakenly regarded as being of neoplastic origin, but are almost always of normal cell origin. Very rarely, however, B-LCL with the same clonal markers as those of neoplastic cells have also been obtained. While the development of B-LCL has been referred to as the in vitro viral immortalization of human B cells and as a phenomenon representing the potential oncogenicity of EBV, the phenotypic and genotypic differences between B-LCL and EBV-carrying BL cells are obvious, indicating that the development of B-LCL per se does not prove the oncogenic activity of EBV. Two EBV-derived antigens, EBNA2 and latent-infection membrane protein (LMP), which are strongly expressed by B-LCL but not by BL cells, have recently been detected in EBV-positive proliferative B cells in patients with organ transplants, suggesting that the proliferating of B-LCL-like cells may take place as an initial step of the multi-step in vivo oncogenesis of EBV.(ABSTRACT TRUNCATED AT 250 WORDS)

B-Lymphocytes

[Suppression of myeloperoxidase gene expression by gamma-interferon].

The gene expression of myeloperoxidase (MPO) in HL60 cells, as tested by Northern blot analysis with a MPO gene probe, was markedly suppressed by treatment with gamma interferon (IFN-gamma)(200U/ml) for 9hr, whereas cytochemically detected MPO activity, cell surface antigen expression, and cell morphology remained unchanged even at 48hr after the treatment. IFN-gamma of 50U/ml was sufficient for the suppression at 24hr. When the HL60 cells treated with IFN-gamma (200U/ml) for 24hr were cultured in the absence of IFN-gamma for another 24hr, the transcript of MPO gene reverted to a level comparable to that of the HL60 cells cultured thoroughly in the absence of IFN-gamma, indicating the reversibility of the suppression. The suppression of MPO expression at RNA level, possibly independent of differentiation, is one of the biological activities exerted by IFN-gamma, which has not been previously reported.

Actins

[Norfloxacin-induced infectious mononucleosis (IM)-like syndrome with Stevens-Johnson syndrome].

A 77-year-old male who had suffered from an upper respiratory infection and had been given Norfloxacin (NFLX) on May 2, 1990, developed generalized erythema which did not subside with prednisolone. He was hospitalized on May 8, and Stevens-Johnson syndrome was diagnosed. The WBC was 115,400/microliter (Ly 61.0%, Aty Ly 39.5%). Sternal tap revealed hypercellular marrow with increased lymphocytes (48.5%; Aty Ly 24.5%) and eosinophils (7.0%). Clinical chemistry revealed slightly abnormal liver and renal function with LDH (1,745 IU/l) and IgE (803 IU/ml) elevation. No pathognomonic result was obtained with several viral antibodies. CD4+, CD8+ lymphocytes and the 4/8 ratio were 39%, 43% and 0.9, respectively. Clinical and laboratory abnormalities were normalized within 3 weeks after the discontinuance of all drugs. Positive lymphocyte stimulation test results were obtained by NFLX. While drug allergy is known to be a cause of IM-like syndrome, there are few reports regarding the subset characterization of the increased T lymphocytes. In this case, T lymphocytosis was remarkable, but the 4/8 ratio declined only slightly, indicating that CD4+ as well as C8+ cells were activated and increased, unlike IM. The record of this case helps to clarify the mechanisms of the lymphocyte activation shared and not shared by EBV-induced IM and IM-like syndrome.

Aged

Long-term respiratory assistance using a right ventricular assist device system and a membrane oxygenator.

A new method for long-term respiratory assistance using a right ventricular assist device (RVAD) system and a membrane oxygenator (MO) was designed and evaluated. An air-driven RVAD was implanted between the right atrium and the pulmonary artery in five goats and an MO was set on the outlet conduit of the RVAD 2 weeks postoperatively. Two types of MOs [the SciMed (Life Systems) silicone sheet MO and a prototype MO using a novel multilayered composite membrane] were used in this study. Respiratory support with this system under systemic heparinization could be conducted for 4-8 days without major problems accompanied with conventional extracorporeal membrane oxygenation (ECMO) such as surgical bleeding, destruction of the blood components, serum leakage, insufficient bypass flow, and fatal systemic thrombosis. This method opened the possibility for ECMO to be used as a bridge before lung or heart-lung transplantation.

Animals

[The absence of monocyte-granulocyte antigen expression in a case of AML].

The brief record of a 25 y. o. male patient with AML (FAB M1) is shown, in whom the blast cells did not express any of the 5 myeloid antigens or the other-lineage related antigens, as detected with the monoclonal antibodies. The blast cells were induced to express CD13 antigen after a short-term culture in vitro. This result suggests that CD13 antigen can be expressed virtually by all AML cells, since CD13 antigen is known to cover fresh AML cells at the highest incidence. No unusual clinical feature was noted in this patient as AML case. The collected documentation of antigen-free AML cases seems necessary for the relevant understanding of AML heterogeneity.

Adult

[The artificial heart: present status and future prospects].

Artificial heart (AH) and heart transplantation (HTx) are complementary means of cardiac assistance and replacement. Temporary use AH including ventricular assist systems (VASs) and total artificial hearts (TAHs) have been put into clinical use. VASs applied to 848 patients in the world and 187 in Japan with acute profound heart failure. The short-term survival rates are 23.3% and 27.3%, respectively. HTx has become an accepted therapeutic procedures. To date, 255 patients received VAS and 182 received TAH as a bridge to HTx, because the number of candidates exceeds the supply of donor hearts. Long-term implantable AH is the future alternatives for cardiac replacement. It is recognized that several key components will be necessary for development of an AH with high performance including biomaterial with antithrombogenecity and durability, an implantable actuator, a noninvasive monitoring system and sensors, a miniature auto-control system, and an energy converter. Another types of devices are on the horizon. These are a muscle-powered biomechanical AH, an AH powered by artificial muscle, and an integrated artificial heart-lung device. With remarkable progress in medicine and engineering, we can expect to see totally implantable permanent use cardiac assist and replacement devices as an alternative for HTx in the near future.

Heart, Artificial

Detection of Epstein-Barr virus genome in benign polyclonal proliferative T cells of a young male patient.

Epstein-Barr virus (EBV) DNA was detected in polyclonal T cells that proliferated transiently in a 21-year-old male (referred to as H.J.) who underwent an apparently benign lymphocytosis (white blood cells, 31 x 10(6)/microL; lymphocyte, 79%) with fever, tonsillar swelling, lymphadenopathy, and hepatosplenomegaly. The symptoms and signs subsided mostly within a month of hospitalization. The major population of the lymphocytes at admission was positive for CD3, CD8 (4/8 ratio, 0.16), WT31, and DR antigen. Eight percent of the leukocytes were too blastoid to be classified as atypical lymphocytes of infectious mononucleosis (IM). The blastoid lymphocytes and the duration and degree of the lymphocytosis and hypergammaglobulinemia appeared inconsistent with IM, whereas the EBV serology indicated either EBV primary infection or a secondary alteration of normal seropositive EBV immunity. The genomic analysis of T-cell receptor beta chain in the peripheral blood mononuclear cells (PBMC) at admission with a C beta probe did not show a monoclonal rearrangement. EBV genome was detected in these cells, using the BamHI W and K probe, but not in the cells after discharge. Analysis of the EBV terminal repeat junctional sequence, using Xho I fragment of the latent membrane protein (LMP) probe binding with the terminus, did not show monoclonal or oligoclonal populations. EBV-associated nuclear antigen (EBNA) was detected in 36% of the PBMC at admission, but not in the later cells. These EBNA-positive cells were found to form rosette with sheep erythrocytes. The PBMC of six acute IM patients contained neither EBV DNA nor EBNA-positive cells. The observations in this case show a unique type of EBV infection in T cells that has not been previously reported.

Adult

Selective expression of the p70 subunit of the interleukin-2 receptor on lymphocytes from patients with infectious mononucleosis.

The lymphocytosis manifested in infectious mononucleosis (IM) during acute phase is ascribed to a reactive expansion of CD8+ T lymphocytes caused by Epstein-Barr virus (EBV)-infected B lymphocytes. Expression of HLA-DR antigen on IM lymphocytes suggests that these T lymphocytes are somehow activated in vivo. In the present study, we analyzed the interleukin-2 (IL-2) receptor expression on lymphocytes from six patients with acute IM. Radiolabeled IL-2 binding assay revealed that IM lymphocytes from all patients examined had a considerable number of IL-2 binding sites with an intermediate affinity, although they did not express the IL-2 receptor recognized by anti-Tac antibody (p55). The number of binding sites (1,070 to 4,600 sites per cell) was larger than that of a normal, resting T lymphocyte-enriched population (650 sites per cell). Furthermore, IM lymphocytes showed marked proliferative responses to higher concentrations of IL-2, which were almost completely blocked by an anti-p70 IL-2 receptor antibody, indicating that their IL-2 receptor is a functional receptor. The results of an affinity cross-linking study seem to indicate that the IL-2 receptor expressed on IM lymphocytes is p70, the second chain of the IL-2 receptor distinct from p55. Flow cytometric analysis following immunofluorescent staining with anti-p70 IL-2 receptor antibody confirmed p70 expression on CD8+ HLA-DR+ lymphocytes. These data suggest that p70 IL-2 receptor expression is involved in the immune response triggered by EBV infection.

Acute Disease

Frequent expression of myeloid antigen (CD13) on immature T cells in culture.

Leukemic cells from 12 patients with lymphoblastic lymphoma (LBL) and T cell acute lymphoblastic leukemia (T-ALL) were studied to determine the inducibility of myeloid antigens in culture in the presence and absence of 12-O-tetradecanoylphorbol-13-acetate (TPA) in association with discrete phenotypic and genotypic analyses on these cells. The investigation revealed that leukemic cells corresponding to common or mature thymocytes were never induced to express any myeloid antigens, and showed rearrangements of T cell antigen receptor (TcR) beta and gamma chain genes. Concomitant examination on leukemic cells from mature T cell malignancies, including adult T cell leukemia (ATL), T cell chronic lymphocytic leukemia (T-CLL) and T cell non-Hodgkin's lymphoma (T-NHL), also failed to express myeloid antigens in culture. By contrast, one of the panmyeloid antigens, CD13 (MCS-2) antigen was induced on leukemic cells corresponding to early thymocytes in 5 out of 7 cases in TPA-added culture and in 3 cases even in TPA-free culture. All of these CD13 antigen inducible cases exhibited the germ line configurations of TcR beta and gamma chain genes except for one case of T-ALL with sole TcR gamma chain gene rearrangement. These findings suggest that primitive T cells, still not undergoing TcR gene rearrangements, retain the characteristics of multipotent progenitor cells to possess different lineage markers and are able to express myeloid antigen not exceptionally. Both phenotypically and genotypically immature thymocytes are considered to be less restricted in the differentiation pathway of hematopoietic cells committed to T cell lineage.

Antigens, Differentiation, Myelomonocytic

[Recovery of severely decompensated hearts with a left ventricular assist device].

We evaluated the limits of recovery of decompensated hearts experimentally and clinically and studied the problems of applying a left ventricular assist device (LVAD). In the chronic experiments, 16 adult goats were studied as follows: Group I consisted of seven with left ventricular infarction in a 70-80% area of the free wall induced by the multiple-ligation method; Group II, three with infarcted areas larger than 80% of the free wall; Group III, three with a 30 min anoxic arrest, and Group IV, a 45-min anoxic arrest. An LVAD was applied in all goats, and a right ventricular assist device was applied simultaneously for two goats in Group IV. Clinically, 21 patients receiving treatment with an LVAD from December 1982 to September 1988 in our center were examined. No goat in Groups II and IV could be weaned from the LVAD. Thus, the severity of artificial heart failure in Groups I and III was considered to be the limit for accomplishing restoration. In successfully-weaned cases, both in experimental and clinical settings, periods of LVAD therapy with or without IABP were less than two weeks, which was thought to be the upper time limits for LVAD application. All but one successfully-weaned patients were alive in the experiment, while clinically many patients died of multiple organ failures regardless of the results of the LVAD assistance. Analysis of data suggested that the difference was caused by the durations of the LVAD applications and weanings, the degree of right ventricular failure, the doses of catecholamine used, and the systemic care afforded, especially during tracheal intubation.

Animals

Ultrastructural analysis of megakaryoblasts by simultaneous detection of platelet peroxidase and platelet glycoproteins.

We studied the phenotype of megakaryoblasts (MKB) in two patients with blastic crisis of chronic myeloid leukemia by simultaneous detection of platelet peroxidase (PPO) activity and platelet glycoproteins (GP) with monoclonal antibodies at the ultrastructural level. When 227A (anti-GPIb) was used, the finding of GP correlated well with the presence of PPO in the majority of MKB, and a few MKB were found to express PPO but lack GP. When 224B (anti-GPIIb/IIIa) was applied, we found a few MKB of a further type, which expressed GP but lacked PPO. These results indicate that the expression of platelet markers is partially aberrant in neoplastic MKB, and that MKB are heterogenous in their phenotypic expressions. Thus, the detection of PPO or GP alone may miss some MKB. For the accurate identification of MKB and the demonstration of aberrant expression of platelet markers, simultaneous detection of the markers seems to useful.

Blast Crisis

Treatment of acute myocardial infarction with cardiogenic shock using left ventricular assist device.

We have treated ten cardiogenic shock patients after acute myocardial infarction (AMI) with a left ventricular assist device (LVAD). These patients were later divided into three groups: the first group with ventricular septal perforation, the second with aorto-coronary bypass grafting (ACBG) before LVAD implantation and the third group without ACBG. LVAD maintained the systemic circulation in each group, and cardiac function recovered enough to remove LVAD in 70% of the total patients. Two of three patients in the first group were discharged from hospital. Two weaned cases in the second group died of multiple organ failure and one was discharged, and hemorrhagic necrosis was seen in the bypassed area of the myocardium. One patient of the third group could not be weaned from LVAD because of respiratory failure though his heart function began to recover. Another case in the third group underwent bypass grafting after removal of LVAD. However ACBG surgery should be done very carefully because a patient in shock is occasionally intolerant to major surgery. In all groups, the major cause of death was multiple organ failure which was probably caused by the prolonged low output condition prior to LVAD application. In the light of this experience, it appears that LVAD should be applied before irreversible damage occurs to major organs, including the heart itself. To ensure the timely application of LVAD, some way must be found to introduce systematic application of LVAD into the normal course of AMI treatment.

Aged