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Biomedical subjects

K Yokoyama

Publications and source records attributed to K Yokoyama.

At least 19 recordsLinked to original sources
PubMed1992

Single site hydrolysis of 2',3'-O-(2,4,6-trinitrophenyl)-ATP by the F1-ATPase from thermophilic bacterium PS3 is accelerated by the chase-addition of excess ATP.

The interaction of 2',3'-O-(2,4,6-trinitrophenyl)-adenosine 5'-triphosphate (TNP-ATP) and TNP-ADP to F1-ATPase from a thermophilic bacterium PS3 (TF1) was investigated. When TNP-ADP or TNP-ATP was added to the isolated alpha or beta subunit of TF1, characteristic difference spectra were generated for each subunit. Difference spectra generated on addition of these analogs to TF1 resembled those observed for the beta subunit, indicating TNP analogs bind to the beta subunits in the molecule of TF1. Results of equilibrium dialysis showed that TNP-ADP binds to a single high affinity site on TF1 in the presence of Mg2+ with a dissociation constant of 2.2 nM. When TNP-ATP was added to TF1 in a substoichiometric molar ratio, it rapidly bound to TF1 and was slowly hydrolyzed. The hydrolysis proceeded nearly to completion without showing stable equilibrium between bound species of TNP-ATP and TNP-ADP. Similar to beef heart mitochondrial F1, this hydrolysis was greatly accelerated by the chase-addition of 100 microM ATP. However, the hydrolyzed product, TNP-ADP, remained bound on the beta subunit even after the chase.

Adenosine Diphosphate↗
PubMed1992

Auditory event-related potential (P300) in relation to peripheral nerve conduction in workers exposed to lead, zinc, and copper: effects of lead on cognitive function and central nervous system.

By measuring auditory event-related potential (P300 and N100) and peripheral nerve conduction velocities, the subclinical effects of lead, zinc, and copper on the central and peripheral nervous system were examined in 22 male gun metal foundry workers exposed to these metals. Their blood lead (BPb) concentrations ranged from 12 to 59 micrograms/dl (median 30). Control subjects were 14 healthy workers, employed at the same factory, who had never been occupationally exposed to these metals. In the gun metal foundry workers, the latencies of P300 and N100 were significantly prolonged; the latency of P300 was significantly correlated with BPb concentrations and other indicators of lead absorption. Similarly, the maximal motor and sensory conduction velocities in the radial and median nerves were significantly slowed, and were significantly correlated with indicators of lead absorption. The data suggest that lead exposure at low levels affects cognitive and central auditory nervous system function together with peripheral nerve conduction.

Adult↗
PubMed1992

Transcriptional regulation of the c-jun gene by retinoic acid and E1A during differentiation of F9 cells.

Differentiation of mouse F9 embryonal carcinoma (EC) cells can be induced by exposure to retinoic acid (RA) or by expression of adenovirus E1A. The transcription of the c-jun gene is stimulated by either RA or E1A. We report here that both RA and E1A strongly induce the expression of chloramphenicol acetyltransferase (CAT) from c-jun promoter/CAT reporter construct (c-jun/CAT), which is stably integrated into F9 cells, in a manner that is independent of both copy number and integration locus. The induction of c-jun/CAT expression is observed in undifferentiated F9 cells, but not in differentiated F9 cells, adenovirus-infected F9 cells or HeLa cells. Deletion analysis of the promoter region of the c-jun gene indicates that the sequence elements required for the RA- and E1A-mediated induction are identical and they have been defined as a region of 145 bp between -190 and -46 of the 5' flanking region of c-jun. This RA and E1A response element (RERE) contains five variants of the motif CGCGGTGACGNT. The upstream two motifs are adjacent and extend in opposite directions, creating an imperfect palindrome. The downstream four motifs are located at 35 or 36 bp intervals in the same orientation. Substitution and insertion analysis indicates that these motifs and their regular intervals are important for the activity of the RERE.

Adenoviruses, Human↗
PubMed1992

Oxidative metabolism of polymorphonuclear leukocytes (PMN) in patients with IgA nephropathy.

The production of hydrogen peroxide (H2O2) by neutrophilic polymorphonuclear leukocytes (PMN) after stimulation and the infiltration of PMN in glomeruli were determined in 20 patients with primary IgA nephropathy. The H2O2 production of PMN after the stimulation was measured with a spectrophotometer using horseradish peroxidase as substrate. The results were as follows: 1) when PMN were pretreated with cytochalasin B, H2O2 production after stimulation with heat-aggregated IgG (IgG) or serum-treated zymosan (STZ) was significantly higher in patients with IgA nephropathy than in controls, and 2) there was an increased amount of PMN localized in glomeruli in patients with IgA nephropathy using immunofluorescence of monoclonal anti-PMN antibody. It appeared that the increased renal infiltration of PMN which have a high potential for production of reactive oxygen species might induce the glomerular injuries in patients with IgA nephropathy.

Adult↗
PubMed1992

Mapping of the human GSPT1 gene, a human homolog of the yeast GST1 gene, to chromosomal band 16p13.1.

The GSPT1 gene, a human homolog of the yeast GST1 gene (formerly named GST1-Hs), was mapped on human chromosome 16p13.1 by a combination of nonradioactive in situ hybridization and Giemsa staining. Southern blot hybridization with a panel of human-rodent somatic cells confirmed the location of the GSPT1 gene on chromosome 16 and also showed the existence of a homologous gene on the X chromosome. A breakpoint for nonrandom chromosome rearrangements has been found in the region of GSPT1 in patients with acute nonlymphocytic leukemia.

Blotting, Southern↗
PubMed1992

Molecular-resolution images of aspirin crystals with atomic force microscopy.

The atomic force microscope was developed in order to image arrays of molecules on the (001), (100) and (011) faces of aspirin crystals in water. Lattice spacings and symmetries of methyl groups and the part of the phenyl groups on the (001) face in water are consistent with X-ray diffraction data. Surfaces of the (100) face that show the best-developed cleavage, in general, are difficult to image. Although the lattice constant derived from (100) image is consistent with the X-ray diffraction data, the c-glide plane symmetry is lost and diffused spots are observed that are 1.8-3.1 times more elongated than (001) spots. It is not possible to image the (011) surface. This initial success in imaging the molecular crystal surfaces of drug compounds has clarified the different submolecular structures of the three different crystal faces, and the close relationship between the imaging difficulties and the differences in the dissolution process in water.

Aspirin↗
PubMed1992

Mapping of the gene family for human heat-shock protein 90 alpha to chromosomes 1, 4, 11, and 14.

The HSP90 family of heat-shock proteins (encoded by genes for HSP90 alpha and beta) constitutes one of the major groups of proteins that are synthesized at increased rates in response to heat and other forms of stress. We previously isolated two distinct cDNA clones for HSP90 alpha from human peripheral blood lymphocytes and from HeLa cells transfected with the adenovirus E1A gene, respectively. To determine the organization of this complex multigene family in the human genome, we used three complementary approaches: Southern analysis of a panel of human/hamster somatic cell hybrids, molecular cloning of the cosmid HSP90 alpha clones from libraries prepared with DNAs from human lymphoblastoid cells, and in situ hybridization to human chromosomes. We demonstrate here that nucleotide sequences that encode HSP90 alpha map to human chromosomes 1q21.2-q22, 4q35, 11p14.1-p14.2, and 14q32.3. The chromosomal mapping of the loci, HSPCAL1, HSPCAL2, HSPCAL3, HSPCAL4, and the characterization of the respective genes should facilitate clarification of the organization of this gene family and lead to a better understanding of the biological functions of the gene product.

Animals↗
PubMed1992

Assessment of slow axonal transport in lead-exposed rats.

To evaluate the effect of lead on slow axonal transport, mean transport velocity (Vmean) of radiolabeled proteins in the motor fibers of the sciatic nerve was calculated by simulating the distribution of radioactivity along the nerve 2 weeks after intraspinal injection of L-[35S] methionine in eight rats given drinking water containing lead acetate for 13 weeks. The transport of individual radiolabeled proteins was also inspected visually by fluorography. It was observed that the Vmean was significantly slowed in the lead-exposed rats and was correlated with blood lead concentration; fluorography indicated that the transport of neurofilament proteins and of tubulins was mildly retarded in the most severely affected lead-exposed rat. These findings suggest that slow axonal transport was impaired by lead. Further study using a large number of animals will be necessary to confirm the findings of the present study.

Animals↗
PubMed1992

Generation of 19 STS markers that can be anchored at specific sites on human chromosome 21.

Sequence-tagged sites (STSs) are short stretches of DNA that can be specifically detected by the polymerase chain reaction (PCR) and can be used to construct long-range physical maps of chromosomal DNA. These STSs can be detected by PCR assays developed by reference to data obtained from the sequencing of restriction fragment length polymorphism-DNA markers for chromosome 21, which were derived from recombinant lamba-phage and plasmid clones made from DNA of a human-hamster hybrid cell line. In this report, we describe the generation of 19 new STSs that are specific for human chromosome 21.

Base Sequence↗
PubMed1992

Development of human monoclonal antibodies against human cytomegalovirus.

Human monoclonal antibodies (HMAbs) against human cytomegalovirus (HCMV) have been developed by fusion of human spleen cells and human lymphoblastoid cell lines (NP101 and NP197). The cell line NP101 had great advantages in its high fusion frequency and the stability of the resultant hybridomas. The specificity of HMAbs was confirmed by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence staining. Two of the six HMAbs obtained, which were IgG3 subclass, neutralized viral infectivity in the absence of complement. The neutralizing activity of one of these two HMAbs was enhanced in the presence of human complement, whereas the other was not. Another IgG1 subclass HMAb neutralized viral infection only in the presence of complement. The remaining three HMAbs showed no neutralizing activity. Those HMAbs may provide an important approach to studying human immune responses to HCMV. HMAbs having neutralizing activity may prove to be useful for passive immunotherapy of HCMV diseases.

Antibodies, Monoclonal↗
PubMed1992

Asymmetric abnormality of renal perfusion with symmetric function in aortic dissection.

Two cases of aortic dissection are presented in which discordance between renal perfusion and function has been demonstrated with Tc-99m DTPA. Renal perfusion on the affected side was decreased on blood flow imaging, whereas renal function, including parenchymal accumulation and excretion assessed by subsequent sequential imaging, were similar for both kidneys. These results suggest that blood flow imaging more readily demonstrates a unilateral renal abnormality in patients with aortic dissection.

Aortic Dissection↗
PubMed1992

Discordant hepatic uptake of Tc-99m NGA and Tc-99m PMT in a patient with hepatoma.

The authors report discordant hepatic uptake of Tc-99m NGA and Tc-99m PMT in a patient with hepatoma. Tc-99m PMT uptake was delayed and Tc-99m NGA concentrated in another area, thereby demonstrating that the uptake mechanisms for Tc-99m NGA and Tc-99m PMT are different. Tc-99m NGA imaging may be useful in characterizing the focal hepatic lesion of Tc-99m IDA or Tc-99m PMT concentration.

Albumins↗
PubMed1992

Tc-99m HMPAO-labeled leukocytes for hematopoietic marrow imaging. Comparison with In-111 chloride.

The current study was initiated to define whether Tc-99m leukocyte imaging, compared to In-111 chloride imaging, could provide additional information on the hematopoietic activity of the bone marrow in diffuse hematologic disorders. A total of 11 patients with hematologic disorders were studied both with Tc-99m leukocytes and In-111 chloride. One patient with myelofibrosis showed disparate tracer distribution in the bone marrow between the two; Tc-99m leukocyte uptake in the central marrow was highly decreased, whereas In-111 chloride showed expanded marrow. The disparate results of granuloid hypoplasia and erythroid hyperplasia obtained by bone marrow biopsy and aspiration in this patient were thought to give an explanation for the radionuclide findings. The remaining 10 patients showed similar tracer distribution in the marrow, and the observation of hematopoietic cellularity confirmed by bone marrow biopsy and aspiration correlated well with the radionuclide findings. These results demonstrate that Tc-99m leukocytes may be a potential imaging agent for evaluating hematopoietic activity in the bone marrow, in particular granulopoietic activity.

Adult↗
PubMed1992

Protective effects of coenzyme Q10 on ischemia-induced reperfusion injury in ischemic limb models.

We examined simple cold preservation of rat limbs in Euro-Collins' solution to elucidate the protective effect of coenzyme Q10 (CoQ10) on the ischemia-induced reperfusion injury in an ischemic extremity replant model. A total of 126 Lewis rat limb replants were performed. Limbs were amputated from donor rats and preserved at 4 degrees C in Euro-Collins' solution and were orthotopically grafted to isogeneic rats by microsurgical technique. In the experimental groups (n = 42), coenzyme Q10 (10 mg/kg) was injected intraperitoneally into the recipients about 1 hour before reperfusion. In the control groups (n = 84), the same dose of solvent was given by the same route. We evaluated vascular patency of anastomoses by direct observation or microangiogram and performed histologic examinations 7 days after replantation. In the control groups, the ischemic limit was 96 hours. Ischemic limbs treated with coenzyme Q10 showed a statistically significant (p < 0.05) improvement in vascular patency after 72 and 96 hours of ischemia. Histologically, bone viability with osteoblastic activity was maintained in coenzyme Q10-treated animals of the 72-hour ischemic group. We conclude that the protective effect of coenzyme Q10 on reperfusion injury is suggested in this replant model.

Animals↗