A new direction in automated laboratory testing in Japan: five years of experience with total laboratory automation system management.

The introduction of integrated laboratory systems has proceeded rapidly in Japan in these 15 years, but they require large initial investment for installation and do not always succeed in reducing laboratory cost. We also experienced three major events that taught us that total laboratory systems are not always effective: these were an earthquake, a nerve gas attack, and an outbreak of food poisoning. Political changes in the national health care system in Japan have forced the cutting of expenses for laboratory testing.

HLA class II polymorphism in Thai patients with non-Hodgkin's lymphoma.

The distribution of HLA-DRB1 alleles and DQB1 alleles in 100 Thai patients with non-Hodgkin's lymphoma (NHL) was analysed using the polymerase chain reaction with sequence-specific primer (PCR-SSP) method, and the association between the disease and the presence of certain HLA class II alleles was investigated. The frequencies of HLA-DRB1*1502 and DRB1*09012 were increased while those of DRB1*0404, DRB1*0803 and DRB1*1106 were decreased. On the other hand, the incidence of HLA-DQB1 alleles was similar to that in the normal population.

Pathophysiological significance of simultaneous measurement of reticulated platelets, large platelets and serum thrombopoietin in non-neoplastic thrombocytopenic disorders.

An automated reticulocyte counter using flowcytometric analysis, the R-3000 (Sysmex Inc. Kobe, Japan), has recently been modified to determine reticulated platelets (RPs) and large platelets (LPs). We measured frequencies of RPs, LPs in total platelet count and serum thrombopoietin concentration comprehensively in non-neoplastic thrombocytopenic patients with immune thrombocytopenic purpura (ITP, n = 23), aplastic anemia (AA, n = 21), liver cirrhosis (LC, n= 17), and hematologically normal subjects (control, n = 151).

[Non-hemophilic patient with inhibitor against factor VIII produced after the second delivery].

A 33-year-old woman who had been healthy and had no history of abnormal bleeding developed widespread ecchymoses and intramuscular bleeding 4 months after her second delivery. On admission, laboratory examination data revealed that factor VIII activity was markedly reduced (4%) and APTT was prolonged (119.7s).

[Recurrent hemolytic uremic syndrome induced by pranoprofen].

A 25-year-old woman was admitted to our hospital because of dark red urine in 1993. A diagnosis of hemolytic uremic syndrome (HUS) because of findings of hemolytic anemia with fragmented erythrocytes, thrombocytopenia, and renal dysfunction. The patient achieved remission with steroids and diuretics.

[PBSCT and GVHD].

Allogeneic peripheral blood stem cell transplantation (PBSCT) has been increasingly used as an alternative to allogeneic bone marrow transplantation. Allo-PBSCT can provide rapid engraftment of neutrophils and platelets. Although the recipients of allogeneic PBSCT are infused 10-fold T cells compared with BMT, there is no evidence for a significant difference between PBSCT and BMT with regard to incidence and severity of acute graft-versus-host disease (GVHD).

[Allogeneic peripheral blood stem cell transplantation].

Allogeneic peripheral blood stem cell transplantation (allo-PBSCT) has been increasingly used as an alternative to allogeneic bone marrow transplantation (allo-BMT). Medication of granulocyte colony stimulating factor (G-CSF) and apheresis are well tolerated by donors and supply adequate numbers of stem cells for the engraftment. Patients engraft sooner using PBSCT compared to allo-BMT.

Microscopic co-distributions of myosin, actin, alpha-actinin and vinculin in human neutrophils during movement.

Human neutrophils initiate their characteristic ameboid movement in response to external stimuli via a mechanochemical system of contractile proteins using mechanical energy derived from hydrolysis of adenosine triphosphate. We have visualized contractile proteins, including myosin, filamentous actin (F-actin), alpha-actinin, tropomyosin, and vinculin in human neutrophils during movement by fluorescence microscopy using double immunofluorescence staining procedures of fluorescein isothiocyanate-labeled antibodies, rhodamine-labeled antibodies, or rhodamine-conjugated phalloidin. From dual color images, both myosin and F-actin, both alpha-actinin and F-actin, both vinculin and F-actin showed co-localization, both myosin and alpha-actinin showed a partial co-localization, at the leading edge in the lamellipodia, and both vinculin and alpha-actinin were co-localized at the leading edge and appeared reciprocally at regular intervals in the lamellipodia.

Assisted reproduction for infertile patients with 9 + 0 immotile spermatozoa associated with autosomal dominant polycystic kidney disease.

We investigated the clinical feature of patients with totally immotile spermatozoa due to 9 + 0 ultrastructural flagellar defects and polycystic kidney disease. We also tried to establish the feasibility of applying modern assisted reproduction technology (ART) in these patients. During 6-year interval a total of 1956 Japanese men were referred to the male infertility clinic.

Automated enumeration of cellular composition in bone marrow aspirate with the CELL-DYN 4000 automated hematology analyzer.

The present study was designed to evaluate the automated analysis of bone marrow aspirates with the CELL-DYN 4000 (CD4000) hematology analyzer. Bone marrow aspirates were diluted twice with phosphate-buffered saline and assayed with the CD4000. The percentages of subpopulations including lymphocytes, neutrophils, and erythroblasts were obtained with the CD4000, and as a reference, differential counts by microscopic observation of May-Grünwald-Giemsa-stained films of bone marrow aspirate were performed (n = 48).

Relative distribution of myosin, actin, and alpha-actinin in adherent monocytes.

The characteristic amoeboid movement of human leucocytes uses mechanical energy derived from the hydrolysis of adenosine triphosphate through a mechanochemical system of the contractile proteins myosin, actin, and the actin-associated protein alpha-actinin. We observed the relative distribution of myosin, actin, and alpha-actinin in adherent monocytes during movement by a double-fluorescence staining procedure. The results indicate that myosin and alpha-actinin are closely associated with the actin cable network, and that alpha-actinin is in close association with the plasma membrane and anchors filamentous actin (F-actin) beneath the plasma membrane; F-actin and alpha-actinin play an important role at the leading edge during the formation of lamellipodia.

Effects of heparin on polymerase chain reaction for blood white cells.

Polymerase chain reaction (PCR) has been used with increasing frequency to diagnose infectious and genetic diseases. In this study, the effects of heparin on PCR were investigated, because heparinized blood may sometimes be used in PCR studies. HLA-DQA1 gene amplification was used as a model.

Limited polymorphism in the HLA-DOA gene.

The HLA-DO molecule, a heterodimer consisting of two novel members of the class II gene family, DOA and DOB, has recently been suggested to function as an important modulator in the HLA class II restricted antigen presentation pathway by interaction with the HLA-DM molecule. In this study, we have analyzed genetic polymorphism and allelic variation of the HLA-DOA gene in 37 HLA class II homozygous typing cells using the direct DNA sequencing technique. As a result, we recognized at least eight allelic variations, DOA*01011, *0101201, *0101202, *0101203, *01013, *0101401, *0101402 and *01015.

Molecular heterogeneity of Krabbe disease.

Krabbe disease (globoid cell leukodystrophy) is an autosomal recessive neurodegenerative disorder that affects both the central and peripheral nervous system due to an enzymatic defect of galactocerebrosidase (GALC). Following its cloning, many mutations in the galactocerebrosidase gene have been reported, but the correlation between phenotype and genotype was not clear in many cases. In this study we further investigated the molecular defects in another 10 patients (6 Japanese and 4 non-Japanese), using cultured skin fibroblasts, and found 10 mutations, of which 8 were novel, including a nonsense mutation (W647X) and 7 missense mutations (G43R, S52F, T262I, Y319C.

[Parvovirus B19 infection-induced anemia and leukocytopenia after myomectomy].

A 38-year woman was hospitalized because of myoma uteri. She underwent myomectomy on September 30, 1997 with 2,000 ml blood loss. No blood transfusion was required, but she received a plasma protein product.

[Detection of anti-IgA alloantibody in a case of non-hemolytic transfusion reaction].

A 59-year-old man with myelodysplastic syndrome exhibited unticaria, dyspnea, fever, and hypotension immediately after the start of platelet concentrate (PC) transfusion on the 15th day of hospitalization. On the 17th and 21st day of hospitalization, the patient exhibited severe unticaria resembling that of erythroderma after the start of PC transfusion. Anti-IgA IgG antibody was detected by enzyme-linked immunosorbent assay (ELISA) on the 28th day of hospitalization.

Klinefelter's syndrome in the male infertility clinic.

The clinical features of patients with Klinefelter's syndrome attending a male infertility clinic have been investigated in order to consider their assisted reproduction treatment options. Over 12 years, a total of 148 patients with sterility due to azoospermia had Klinefelter's syndrome. Eight patients were shown by fluorescence in-situ hybridization (FISH) on metaphase spreads to be mosaic (46,XY/47,XXY), and 140 patients showed only 47,XXY.

Which type of underlying disease facilitates cytomegalovirus infection? Comparison of benign disease, hematopoietic malignancy, and post-bone-marrow or renal transplantation status by using the first standardized objective PCR test for cytomegalovirus detection.

All immunocompromised hosts, such as infants, the elderly, patients with advanced cancer, and patients being treated with immunosuppressants, etc., are said to be more susceptible to cytomegalovirus (CMV) infection or CMV disease. However, we questioned the validity of this conclusion and attempted to detect CMV viremia in the plasma of subjects by using the AMPLICOR CMV test (Roche Diagnostics Systems, Branchburg, NJ), the first standardized PCR kit for CMV infection.

Clinical usefulness of conjugated bilirubin levels in patients with acute liver diseases.

We used a specific method to measure conjugated bilirubin levels in patients with acute liver diseases to examine its clinical usefulness. Conjugated and total bilirubin levels were measured in 102 samples obtained from six patients with acute liver diseases (three with fulminant hepatic failure, one with acute severe hepatitis and two with acute hepatitis; see text for criteria). Total and conjugated bilirubin levels were measured with Iatro T (total)-Bil and D (direct)-Bil kits (Iatron Laboratories Tokyo, Japan) and with conventional Nescauto T(total)-Bil and D(direct)-Bil VE kits (Nippor Shoji, Osaka, Japan).

[Human parvovirus B19-induced aplastic crisis in a patient treated with fibrin sealant].

A 42-year-old woman underwent a myomectomy on March 31, 1998. On the 10th postoperative day, leukopenia and reticulocytopenia were observed. Bone marrow aspiration revealed severe erythroblastopenia with giant proerythroblasts, suggesting a recent parvovirus infection.

A comparative study of administration methods of granisetron injection used to treat nausea/vomiting induced by cancer chemotherapy without cisplatin in tumors of hematopoietic organs. Keihanshin Study Group of Hematological Malignancies.

Purpose: The antiemetic effect of granisetron injection at a dose of 40 microg/kg used in the treatment of nausea/vomiting induced by multidrug combined cancer chemotherapy excluding cisplatin in patients with tumors of hematopoietic organs was evaluated by comparing a 30-min infusion and a slow intravenous injection given over 30 s.

Methods: A two-group random-allocation comparative study was performed with the cooperation of multiple institutions using a central registration system.

Results: In the treatment of acute clinical symptoms, appetite was described as "similar to that during good health" by 61.

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.