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Biomedical subjects

K Shimura

Publications and source records attributed to K Shimura.

At least 37 records · Page 2Linked to original sources

[CD5 positive B cell leukemic lymphoma associated with BCL6 rearrangement].

A 59-year-old man was admitted in December 1995 because of general fatigue without lymphadenopathy. Increased abnormal lymphocytes (70%) were observed in peripheral blood. Bone marrow aspiration was a dry tap. Biopsy specimens revealed hypercellularity with infiltration of abnormal lymphocytes. Surface marker analysis of tumor cells was positive for CD5, CD19, CD20, HLA -DR, kappa, and sIgM and negative for CD10. Cytogenetic analysis disclosed a complex abnormal karyotype including t(3;22) and rearrangement of the BCL6 gene. The patient was given a diagnosis of CD5 positive B-cell lymphoma, but died in January 1997 despite repeated chemotherapy. This case was unique because BCL6 rearrangement has been reported in various types of B-cell lymphoma but rarely associated with leukemic types without lymphadenopathy.

CD5 Antigens↗

Mouse alloantigen Ly10 is identical to murine fusion regulatory protein-1 (mFRP-1)/4F2/CD98: aberrant expression of mFRP-1/Ly10 allotypes in cells derived from CDF1 mice due to the gene deletion.

Murine FRP-1 (mFRP-1) and Ly10 antigens are alloantigens. Anti-Ly10.1, monoclonal antibody (mAb) reacts to HeLa cells stably expressing mFRP-1.1 (HeLa cells/mFRP-1.1), but does not react to HeLa cells/ mFRP-1.2. On the other hand, anti-Ly10.2 mAb reacts to HeLa/mFRP-1.2 cells, but does not react to HeLa cells/mFRP-1.1. These findings indicate that Ly10.1 and Ly10.2 antigens are identical to the heavy chains of mFRP-1.1 and mFRP-1.2 molecules, respectively. Thymocytes and fibroblast cells obtained from CDF1 and BDF1 mice showed reactivity to both anti-Ly10.1 and anti-Ly10.2/FRP-2 antibodies, indicating that mFRP-1/Ly10 alloantigens are codominantly expressed in the F1 mice. Intriguingly, IMC carcinoma cells, derived from CDF1 mice, express mFRP-1.2, but do not express mFRP-1.1. Surprisingly, DBT cells, derived from CDF1 mice, consist of two different cell populations: one expresses both mFRP-1.1 and mFRP-1.2 antigens and the other expresses only mFRP-1.1 antigen. Consequently, we tried to isolate cloned DBT cell lines by limiting dilution. Finally, six cloned DBT cell lines were obtained, and three clones of these cloned DBT cells expressed both mFRP-1.1 and mFRP-1.2, and another three clones of these cloned DBT cells expressed both mFRP-1.1 and mFRP-1.2, and another three clones including clone 5 cells expressed only mFRP-1.1 antigen. No induction of mFRP-1.2/Ly10.2 by 5-azacytidine and sodium n-butyrate could be detected in DBT clone 5 cells. Neither mFRP-1.2 mRNA nor the genomic cDNA clone encoding mFRP-1.2 could be detected in DBT clone 5 cells, nor could the genomic cDNA encoding mFRP-1.1 be detected in IMC cells, indicating that aberrant expression of mFRP-1/Ly10 allotypes in CDF1 mice-derived cells is due to the gene deletion.

Animals↗

Capillary affinophoresis as a versatile tool for the study of biomolecular interactions: a mini-review.

Combination of capillary electrophoresis and bioaffinity interaction gave rise to powerful research tools for analyzing molecular recognition. They take advantages of the electrophoretic behavior of the complex formed between a target biomolecule and a specially designed mobile ligand molecule (affinophore or affinity probe), and enable detection of complex formation, determination of the equilibrium constants and stoichiometry, etc.

Affinity Labels↗

Capillary affinophoresis of pea lectin with polyliganded affinophores: a model study of divalent-polyvalent interactions.

Affinophoresis is a type of affinity electrophoresis using an affinophore, a soluble ionic carrier bearing affinity ligand(s). It was reported previously that an affinophore, prepared by coupling multiple p-aminophenyl alpha-D-mannoside ligands to a part of the carboxyl groups of succinylated polylysine, specifically changed the mobility of pea lectin in agarose gel. The affinophoresis of this divalent lectin with the polyliganded affinophore was investigated by using capillary electrophoresis. Analysis of the mobility change of the lectin in the presence of differently modified affinophores showed that the affinity was larger for affinophores having higher ligand density. Analysis of the inhibition of the mobility change by a neutral ligand, with a known affinity constant for the lectin, allowed estimation of the contributions of monovalent and divalent interactions to the binding in the lectin-affinophore complex. The proportion of divalent complexes was greater for affinophores having higher ligand density. This approach to estimate the contribution of divalency in complex formation should be generally applicable to the analysis of divalent interactions with different techniques other than electrophoresis.

Aniline Compounds↗

Assay of trypsin activity by capillary isoelectric focusing with laser-induced fluorescence detection.

Capillary isoelectric focusing is a highly effective method for the separation of proteins due to focusing as a function of their pI values in the separation process. This technique is also effective for certain types of peptides that focus well. Fluorescence labeling and subsequent detection by laser-induced fluorescence farther enhance the sensitivity of this technique. This paper demonstrates the utility of this technique in an enzyme assay. A synthetic nona peptide, H-Gly-Cys-His-Glu-Ala-Arg-Ala-Glu-Glu-OH, was labeled with an iodoacetyl derivative of Lissamine rhodamine B at the thiol group of the cysteine residue as a substrate for trypsin. Trypsin catalyzed the cleavage of the Arg-Ala bond of the labeled substrate, which focused at pH 4.8, and liberated a shortened, labeled product, H-Gly-*Cys-His-Glu-Ala-Arg-OH that focused at pH 6.9 (* indicates the label). The product peptide at 3-300 pM was determined with a relative standard deviation of 5.5% (n = 5) by fluorescence detection at 590 nm with excitation by a green line of He-Ne laser. Incubation of trypsin with the substrate for 10 min at 37 degrees C allowed the determination of 50-250 pg of trypsin, with a relative standard deviation of 5.3% (n = 5).

Amino Acid Sequence↗

Flow cytometric analysis of Thy-1 expression in myelodysplastic syndrome.

We analyzed the expression of Thy-1 (CD90) antigen on CD34+ bone marrow mononuclear cells (BMMNC) obtained from 25 patients with myelodysplastic syndrome (MDS) by two-color flow cytometry. Five of nine patients (55.6%) with refractory anemia with excess of blasts (RAEB) and two of 16 (12.5%) with RAEB in transformation (RAEB-t) showed more than 20% of Thy-1 expression of their CD34+ BMMNC. Regarding chromosomal abnormalities, -5/5q- or -18 might be correlated with the expression of Thy-1 on CD34+ BMMNC in MDS. Nine patients were analyzed twice, once before and once after leukemic transformation and showed no significant change in Thy-1 expression. These results show that Thy-1 was expressed on CD34+ BMMNC in certain patients with MDS before leukemic transformation and that it was maintained during the disease progression. In contrast, expression of Thy-1 did not seem essential to the leukemic transformation in MDS though the patients with high Thy-1 expression might have poorer prognosis compared with those with low Thy-1 expression.

Adolescent↗

Detection of endogenous retrovirus antigens in NOD mouse pancreatic beta-cells.

We characterized C-type retroviruses expressed in the pancreatic beta-cells of non-obese diabetic (NOD) mice by immunohistochemical techniques and by inhibiting the production of viral particles using antisense oligonucleotides. Some cells in the pancreatic islets from both NOD and diabetes-resistant NOD-related mice (NON) reacted with a monoclonal antibody directed against the envelope protein(s) of polytropic viruses. On the other hand, NOD islet cells also showed strong immunoreactivity with an anti-gag protein monoclonal antibody and another anti-envelope protein(s) monoclonal antibody that is specific for xenotropic viruses. In antisense oligodeoxynucleotide inhibition assays, a xenotropic virus-specific phosphorothionate-particles antisense oligodeoxynucleotide significantly inhibited the occurrence of C-type virus particles in NOD mouse islet beta-cells. Therefore, C-type retrovirus-like particles expressed in NOD mouse pancreatic beta-cells were considered to be endogenous xenotropic virus. The expression of the xenotropic viral genome may be involved in the pathogenesis of the diabetic syndrome in NOD mice.

Animals↗

Sarcocystis suihominis detected for the first time from pigs in Japan.

Sarcoystis suihominis was detected for the first time in Japan from the heart and diaphragm of 5 out 600 older culled breeding pigs slaughtered in Saitama Prefecture, Japan. Fresh cysts were 1,080-2,040 x 106-170 microns in size. Bradysoites measured 15 x 4 microns on average. The cyst wall was usually observed thick, 4-6 microns, and striated, but occasionally thin and smooth according to the difference in sectioning angle and in portion of cysts. Scanning electron microscopy showed that many palisade-like villar protrusions, 6-6 x 0.3-0.5 microns in size, were closely folded onto the surface of cyst. A small number of microtubules were seen in the core of protrusion. No dogs nor domestic cats fed with 20 fresh cysts each excreted oocysts or sporocysts in the feces throughout the experimental period of 30 days.

Animals↗

Effects of spin labeled derivatives of podophyllotoxin on cell cycle and macromolecular synthesis in human lymphoid leukemia Molt 4B cells.

AIM: To examine the effect of the spin labeled derivatives of podophyllotoxin, N-podophyllic acid-N"-[4-(2,2,6,6-tetramethyl-1-piperidinyloxy)] thiosemicarbazide (GP4) and 4-[4"-(2",2",6",6"-tetramethyl-1"-piperidinyloxy) amino]-4'-demethylepipodophyllotoxin (GP7) on the cell cycle and macromolecular synthesis of human lymphoid leukemia Molt 4B cells in vitro. METHODS: MTT assay, 3H incorporation, and flow cytometer were used. RESULTS: GP4, GP7, and etoposide 0.02-100 mmol.L-1 cultured for 48 h inhibited the proliferation of human lymphoid leukemia Molt 4B cells. IC50 values of GP4, GP7, and etoposide were 0.11, 4.7, and 1.6 mmol.L-1, respectively. DNA and protein syntheses were obviously suppressed by GP4, GP7, and etoposide 10 mmol.L-1 for 48 h. After Molt 4B cells were treated with GP4, GP7, and etoposide 10 mmol.L-1 for 6 and 12 h, the mitotic index was increased by GP4 and reduced by GP7 and etoposide. According to flow cytometric BrdU/DNA analysis, GP4 slightly retarded S phase and mainly arrested cell cycle progression in G2/M phase, whereas GP7 similar to etoposide induced cells accumulated at S phase and retarded the cells in G2 phase. CONCLUSION: GP4 and GP7 inhibit the proliferation of Molt 4B cells, but the mechanisms are different.

Antineoplastic Agents, Phytogenic↗

Unsharp masking technique using multiresolution analysis for computed radiography image enhancement.

An unsharp masking technique (USM) is one of the image processing methods used in the computed radiography (CR) system. To further promote the performance of the USM, we developed the NEW-USM processing that can control the frequency enhancement characteristics flexibly and accommodate an extensive range of diagnostic targets. The NEW-USM and USM were performed on femur images acquired by computed radiography (model FCR9000; Fuji Medical Systems USA Inc, Stamford, CT), and the resulting images were compared. In the NEW-USM image, bone structures are enhanced as sharply as in the USM image, whereas the surrounding soft tissue structures, such as muscle are enhanced more strongly than in the USM image. Furthermore, the absence of the bone structure that may suggest pathological change is more obvious in the NEW-USM image. The newly developed NEW-USM can appropriately enhance diagnostic information over the whole range of image frequencies, thereby expanding utility of the USM.

Algorithms↗

Discrimination of eight chicken Eimeria species using the two-step polymerase chain reaction.

A method was developed for the discrimination of 8 Eimeria species of chickens, i.e., E. acervulina, E. brunetti, E. mitis, E. maxima, E. necatrix, E. praecox, E. tenella, and E. hagani using the 2-step polymerase chain reaction (PCR). In the first PCR, the small subunit ribosomal RNA (srRNA) gene was amplified from the parasite genome using conserved sequences for the Apicomplexa srRNA gene as the primers. The srRNA gene amplified from the parasite genome was discriminated in the second step by random-amplified polymorphic DNA (RAPD) PCR using 10 arbitrary primers. Each arbitrary primer produced species-specific RAPD patterns that provided a simple method for species identification from the srRNA genes of the 8 Eimeria species. This method should be useful for discrimination of the parasite species for diagnosis or epidemiological surveys of chicken coccidiosis.

Animals↗

[A case of IAHS (infection associated hemophagocytic syndrome) successfully treated with etoposide].

This report details a case of infection associated hemophagocytic syndrome (IAHS). A 20-year-old female was admitted to our hospital with persistent high fever in July, 1994. Physical examination revealed high body temperature (40 degrees C), marked hepatosplenomegaly and no superficial lymph node swelling. Laboratory examination revealed leukopenia and abnormal liver function on admission. Serum ferritin levels were surprisingly elevated. The coagulation tests showed high FDP and D-dimer. Specific viral antibody titers were not elevated such as Epstein-Barr virus or Cytomegalovirus. Bone marrow examination revealed histiocytic hyperplasia with hemophagocytosis, and the histiocytes were well matured. We diagnosed IAHS. Corticosteroids were administered on the 3rd hospital day (methylprednisolone 1 g/day, 3 days), but persistent high fever and laboratory findings did not improve. So we tried etoposide (etoposide 200 mg/day, 5 days) therapy on the 13th hospital day. After administration of etoposide, she failed to recover from severe leukopenia and suffered from meningitis. We administered G-CSF, gamma-globulin and antibiotics for intensive supportive therapy. As the leukocyte count increased, her symptoms and laboratory data improved. There was no hemophagocytosis in her bone marrow before discharge. Recently, etoposide is said to be effective for reactive monocytic proliferation. Administration of etoposide was very effective for IAHS, although corticosteroids, were ineffective.

Adult↗

Determination of the affinity constants of pea lectin for neutral sugars by capillary affinophoresis with a monoligand affinophore.

Affinophoresis is a type of affinity electrophoresis in which an affinophore, a conjugate of an affinity ligand and a multiply charged soluble matrix, causes a change in migration velocity of proteins which have a specific affinity for the ligand. A monoligand affinophore bearing a mannoside was prepared by coupling iodoacetylated p-aminophenyl alpha-D-mannoside to the free thiol group of N-succinylated glutathione, and used for the affinophoresis of pea lectin in a capillary. The electrophoretic mobility of pea lectin towards the anode increased in the presence of the affinophore as a function of its concentration in a manner that is described by the equation for affinity electrophoresis. Analysis of the suppression of the affinophoresis on the addition of neutral sugars to the system allowed the determination of the dissociation constants of the lectin for these neutral sugars. The dissociation constants obtained on affinophoresis agreed well with the values in the literature. The preparation of a monoligand affinophore for ligands bearing an amino group should facilitate the application of this type of microscale analysis (0.14 ng of protein for each run) to protein ligand interactions.

Affinity Labels↗

An increase in cortical excitability with no change in spinal excitability during motor imagery.

During motor imagery, to estimate changes in excitability of flexor carpi radialis muscle motoneurons of the spinal and cortical levels, electrical stimuli for recording H-reflex and transcranial magnetic stimulation (TMS) for recording motor evoked potentials (MEPs) were used. In the absence of movement or detectable EMG activity during motor imagery, there was an increase in cortical excitability with no change in spinal excitability.

Adult↗