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Biomedical subjects

M Ono

Publications and source records attributed to M Ono.

At least 505 records · Page 28Linked to original sources
PubMed1994

Protective effect of individual glycoproteins of Newcastle disease virus expressed in insect cells: the fusion protein derived from an avirulent strain had lower protective efficacy.

Recombinant hemagglutinin-neuraminidase (rHN) and fusion (rF) glycoproteins of virulent and avirulent strains of Newcastle disease virus (NDV) expressed by using baculovirus expression system were used to investigate their protective immunization effects in chickens. The efficacy of immunization with these recombinant proteins was evaluated by challenge infection. The chickens immunized with either rHN or rF protein of a virulent strain or rHN protein of an avirulent strain were completely protected from the lethal infection of virulent NDV. On the other hand, the rF protein of an avirulent strain, in which precursor F protein was not cleaved, showed lower protective effects. Significant levels of specific antibodies against respective proteins were detected in sera from survivors, whereas relatively lower levels of antibodies were found in chickens which were killed by challenge infection. These data indicate that either HN or F protein alone could induce protective immune responses and the cleavage of F protein might be important for its immunological potential.

Animals↗
PubMed1994

External pH modifies sodium channel block by mexiletine in guinea pig ventricular myocytes.

OBJECTIVE: The aim was to examine the kinetics and the degree of Na+ channel block by mexiletine under different values of external pH to produce a fractional variation of charged and uncharged forms of mexiletine. METHODS: The Na+ current (INa) was recorded using the patch clamp technique of whole cell configuration in guinea pig ventricular myocytes. RESULTS: In the presence of 20 microM mexiletine, INa block developed with a single exponential when depolarising pulse trains of 5 ms were applied to -30 mV from a holding potential of -100 mV at a diastolic interval of 500 ms. At the 30th pulse, the degree of use dependent block of INa was 12.3(SEM 0.41)% (n = 4) of the 1st pulse. When the duration of pulses was prolonged to 200 ms, the degree of use dependent block was increased to 25.6(4.7)%, which had the block development of two exponentials with fast and slow rate constants. The slow component at the 200 ms pulses was comparable to the single exponential component at the 5 ms pulses. Drug binding to the inactivated channel was increased by external alkalinisation, which increased the uncharged form of mexiletine, whereas the block at the 5 ms pulses was not affected by the change of external pH. CONCLUSIONS: Uncharged mexiletine caused use dependent block with a fast rate constant having affinity for the inactivated state of the Na+ channel, while the charged form produced a slow component of the block having affinity for the activated state. Therefore the fraction of the uncharged to the charged form of mexiletine could be one of the important determinants in the development and mechanism of INa block.

Animals↗
PubMed1994

Three different types of dry eye syndrome.

We analyzed patients with dry eye syndrome with regard to autoimmune conditions. A total of 116 patients with dry eye syndrome were divided into three groups: simple dry eye (SDE), i.e., dry eye with no circulating autoantibodies; autoimmune positive dry eye (ADE), dry eye with circulating autoantibodies; and Sjogren's syndrome (SS), dry eye associated with Sjogren's syndrome. Schirmer test showed values of 3.0 +/- 2.2 mm in SDE, 3.1 +/- 2.0 mm in ADE, and 2.4 +/- 2.3 mm in SS reflecting the inadequacy of this test in differentiating among the groups. However, Schirmer test with nasal stimulation showed values of 19.1 +/- 12.4 mm in SDE and 16.4 +/- 10.9 mm in ADE, which were significantly higher than the 7.0 +/- 6.6 mm found in SS (p < 0.01). Moreover, ocular surface alterations evaluated by vital staining and brush cytology were significantly milder in SDE and ADE than in SS. SDE and ADE have less ocular surface abnormalities with good reflex tearing, whereas SS has less reflex tearing and more squamous metaplasia.

Adult↗
PubMed1994

Inhibition of feline immunodeficiency virus gene expression and replication by alphaherpesvirus ICP4 homologues.

We investigated the effects of pseudorabies virus (PRV), herpes simplex virus type 1 (HSV-1), and equine herpesvirus type 1 (EHV-1) ICP4 homologues on feline immunodeficiency virus (FIV) long terminal repeat (LTR)-directed gene expression. This was done by using the transient expression chloramphenicol acetyltransferase (CAT) assay in Crandell feline kidney (CRFK) and Felis catus whole fetus 4 cells transfected with a chimeric FIV LTR-CAT reporter construct in combination with effector plasmids expressing the PRV, HSV-1 or EHV-1 ICP4 homologue. The experiments demonstrated that the ICP4 homologues could significantly inhibit FIV LTR-directed gene expression. Moreover, the ICP4 homologues also exhibited a marked inhibitory effect on FIV replication in CRFK cells cotransfected with an infectious molecular clone of FIV.

Animals↗
PubMed1994

Cooperative roles of hepatocyte growth factor and plasminogen activator in tubular morphogenesis by human microvascular endothelial cells.

Epidermal growth factor (EGF) or transforming growth factor-alpha (TGF-alpha) stimulated cell migration, chemotaxis, and the expression of tissue-type plasminogen activator (t-PA) in human omental microvascular endothelial (HOME) cells. Hepatocyte growth factor (HGF) stimulated cell proliferation, but had a negligible stimulatory effect on cell migration, the expression of t-PA and tube-like formation into collagen gel in HOME cells. Basic fibroblast growth factor stimulated cell proliferation, cell migration, tubulogenesis and the expression of urokinase-type plasminogen activator (u-PA) in bovine aortic endothelial (BAE) cells. HOME and BAE cells had both high- and low-affinity receptors for HGF. In BAE cells, u-PA activity and tube-like structures in collagen gel were induced in the presence of HGF alone. In contrast, in HOME cells, t-PA activity and tube-like structures were induced in the presence of TGF-alpha alone, but not in the presence of HGF alone. However, we observed a marked induction of tube formation by HOME cells when both t-PA and HGF were added simultaneously. In the model system for tumor angiogenesis, when HOME cells were co-cultured with a renal cancer cell line, KPK13, tube-like structures were induced in the presence of HGF:KPK13 cells expressed large amounts of t-PA mRNA. Our present study suggested that HGF in concert with active t-PA could be angiogenic in HOME cells.

Cell Line↗
PubMed1994

Characterization and localization of mouse hypothalamic growth hormone-releasing factor and effect of gold thioglucose-induced hypothalamic lesions.

Hypothalamic growth hormone-releasing factor (GRF) in higher mammals, including human GRF, is a 44 amino acid residue peptide and is highly homologous in structure. By contrast, mouse GRF (mGRF) recently deduced by cDNA cloning consists of only 42 residues and shows relatively low homology to the GRFs of higher mammals and the same rodent species, rat. To characterize and localize the predicted mature mGRF peptide in the hypothalamus, we have generated its antiserum and developed a homologous radioimmunoassay. Immunoreactive mGRF in the acid hypothalamic extract was eluted as a single peak at a position identical to that of synthetic peptide on both gel filtration chromatography and reverse-phase high-performance liquid chromatography (HPLC). Secretion of immunoreactive mGRF from incubated hypothalami increased several fold in response to 50 mM K+, and this rise was abolished in the absence of medium Ca2+. Only a single peak of immunoreactive mGRF that coeluted with synthetic replicate was observed after the K(+)-stimulated medium was extracted on Bond Elut C18 cartridges and applied on reverse-phase HPLC. Immunohistochemistry identified many mGRF-positive cell bodies in the arcuate nucleus and dense bundles of immunoreactive fibers in the median eminence. Treatment of mice with gold thioglucose (GTG), a chemical agent known to cause hypothalamic lesions, markedly depleted both content and in vitro secretion of immunoreactive mGRF. The decline in mGRF secretion was greater in GTG obese than in nonobese mice, whereas somatostatin secretion was not affected by GTG treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

N-methyl-D-aspartate induces phase shifts in circadian rhythm of neuronal activity of rat SCN in vitro.

The suprachiasmatic nucleus (SCN) acts as a pacemaker for mammalian circadian rhythms. Receptors for excitatory amino acids like N-methyl-D-aspartate (NMDA) and non-NMDA receptors have both been found to play an important role in the transmission of photic information from the retina to the SCN. Therefore, we investigated whether the application of glutamate receptor agonists could reset the phase of the circadian rhythm of SCN firing activity in vitro. Treatment with NMDA (0.1-10 microM) for 15 min or 1 h during the early part of the subjective night produced phase delay, whereas treatment during the late subjective night caused an advance in phase. The phase-response curve for NMDA was similar to that previously obtained in response to light pulses in vivo. Application of DL-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid hydrobromide (AMPA) (1 or 10 microM), a non-NMDA-receptor agonist, also produced a dose-dependent phase delay of SCN activity. The NMDA-induced phase delay was antagonized by an NMDA-receptor antagonist MK-801. These findings suggest that both NMDA and non-NMDA receptors may be involved in the transmission of information to the SCN via the retinohypothalamic tract. In addition, both the advances and delays in phase caused by NMDA were potentiated by cotreatment with neuropeptide Y, whereas AMPA-induced phase delay was not potentiated by neuropeptide Y. This points to a functional link between NMDA and neuropeptide Y receptor-mediated mechanisms in the SCN.

Animals↗
PubMed1994

Pit-1/GH factor-1 involvement in the gene expression of somatolactin.

A presumed pituitary hormone, somatolactin (SL), belonging to the GH/PRL family, is produced in the intermediate lobe of teleost pituitary. The pituitary-specific transcription factor, Pit-1, is indispensable not only for the expression of mammalian GH and PRL genes, but for the development of GH- and PRL-producing cells present in the anterior lobe of the pituitary as well. Thus, in this study, examination was made of the possible involvement of Pit-1 in the intermediate lobe-specific expression of the SL gene. Using antibodies against chum salmon Pit-1, the presence of the 40-kilodalton Pit-1 protein in anterior and neurointermediate lobes of rainbow trout pituitary was demonstrated. By immunohistochemical examination, Pit-1 protein was shown to be located in the nuclei of SL-producing cells in the intermediate lobe. In experiments of cotransfection into HeLa cells, rat Pit-1 enhanced the promoter activity of SL gene, and a 0.5-kilobase upstream region from the transcriptional start site was sufficient for this enhancement. It follows from these results that Pit-1 protein may possibly be involved in SL gene expression as well as the development of SL-producing cells.

Animals↗
PubMed1994

The heparin-binding domain of heparin-binding EGF-like growth factor can target Pseudomonas exotoxin to kill cells exclusively through heparan sulfate proteoglycans.

Heparin-binding EGF-like growth factor (HB-EGF) is a smooth muscle cell mitogen composed of both EGF receptor and heparin-binding domains. To better understand the function of its domains, intact HB-EGF or its heparin-binding (HB) domain (amino acids 1-45) were fused to a mutant Pseudomonas exotoxin with an inactivated cell-binding domain. The resulting chimeric toxins, HB-EGF-PE* and HB-PE*, were tested on tumor cells, proliferating smooth muscle cells and a mutant Chinese hamster ovary cell line deficient in heparan sulfate proteoglycans (HSPGs). Two targets were found for HB-EGF-PE*. Cells were killed mainly through EGF receptors, but the HB domain was responsible for killing via HSPGs. HB-PE* did not bind to the EGF receptor and thus was cytotoxic by interacting exclusively with HSPGs. We conclude that the HB domain of HB-EGF is able to mediate internalization through HSPGs, without requiring the EGF receptor.

ADP Ribose Transferases↗
PubMed1994

Microtubule reorganization and lysosome redistribution by a viral v-src oncogene, in mouse Balb/3T3 cells expressing human EGF receptor.

The epidermal growth factor (EGF)-induced endocytosis of its receptor is an obligatory pathway for the cellular regulation of the EGF-specific receptor (EGF-R). BNER4 is a mouse Balb/3T3 cell line transfected with human EGF-R complementary DNA (cDNA). B4/src-13 and B4/src-24 are BNER4 cells transfected with a viral oncogene v-src. Indirect immunofluorescence study demonstrated that EGF-R was mostly localized at the perinuclear region in BNER4 cells at 60 min after EGF addition, whereas it was diffusely distributed throughout the cytoplasm in its v-src transfectants. Double indirect immunofluorescence study further confirmed that EGF-R was localized in lysosomes in BNER4 and B4/src-13 cells at 60 min after EGF addition. Intracellular distribution of the Golgi apparatus, clathrin-coated vesicles and early endosomes were similar in all cell lines. However, the lysosomes detected by anti-lysosomal membrane protein (LGP85) antibodies were diffusely distributed throughout the cytoplasm in the v-src transfectacts. By contrast, in the parental BNER4 cells, the lysosomes were mostly localized in the perinuclear region. The organization of microtubules, but not of actin, was markedly different between BNER4 cells and its v-src transfectants. Nocodazole, which depolymerizes microtubules, altered the distribution of the lysosomes and EGF-R in BNER4 cells. Both intracellular lysosome distribution and microtubule organization in nocodazole-treated BNER4 cells were found to be similar to those in its v-src transfectants without nocodazole treatment. These findings support the notion that changes in lysosome distribution may be correlated with microtubule reorganization by v-src in mouse Balb/3T3 cells.

3T3 Cells↗
PubMed1994

Case report: splenic vein aneurysm--report of a lesion that progressively expanded.

A case of splenic vein aneurysm is reported. The patient was diagnosed as having a cirrhotic liver with portal hypertension. Computed tomography and angiography demonstrated a splenic vein aneurysm with saccular dilatation. The splenic vein aneurysm increased in size as the hepatic cirrhosis deteriorated; however, there were no complications such as rupture or thrombosis. To our knowledge this is the first report describing the development of a portal system aneurysm.

Aneurysm↗
PubMed1994

Nephritogenic antibodies in MRL/lpr lupus mice: molecular characteristics in pathological and genetic aspects.

MRL/lpr mice spontaneously develop a lethal glomerulonephritis (GN). We found that IgG3 production in this strain of mice has a critical role on the development of GN; 1) IgG3 levels were high in kidney-extracted IgG and in circulating IgG immune complexes (IC), 2) serum IgG3 was selectively reduced by cyclosporin A treatment, associated with amelioration of GN, and 3) the mRNA levels of IgG3 correlated well with the severity of GN among the MRL/lpr x (MRL/lpr x C3H/lpr) F1 backcross mice with the rearranged genetic profile. Based on these results, we have successfully established five hybridoma clones which produce nephritogenic IgG3 antibodies from an unmanipulated MRL/lpr mouse. When they were injected to normal mice, four of the five clones generated cell-proliferative GN associated with the marked cellular infiltrates, while the remaining clone induced wire loop-like lesions. This result suggests that particular antibodies generated in MRL/lpr mice have a different pathogenic potency. The V-region sequence study of these nephritogenic antibodies revealed that the two types of the glomerular lesions were mediated by a different B cell precursor. In conclusion, GN in MRL/lpr lupus mice is thought to be generated by the expansion of clonally different B cells producing nephritogenic antibodies with a different pathogenic potency.

Amino Acid Sequence↗
PubMed1994

Pituitary-specific transcription factor Pit-1 in the rdw rat with growth hormone- and prolactin-deficient dwarfism.

The pituitary gland of the rdw rat (gene symbol: rdw) with hereditary dwarfism expresses 30-100 times less GH and prolactin (PRL) mRNA than normal controls. To clarify the features of rdw rats, TSH and the pituitary-specific transcription factor Pit-1, which is involved not only in the gene expression of GH and PRL but in somatotroph, lactotroph and thyrotroph development as well, were examined. The rdw pituitary contained about seven times more TSH beta mRNA than the normal control, whereas Pit-1 mRNA expression in rdw and control was the same. Nucleotide sequencing of PCR-amplified Pit-1 cDNA indicated that the deduced amino acid sequence of rdw Pit-1 was identical with that of the normal rat. Using an antibody against rat Pit-1 protein produced in E. coli, Western blotting analysis demonstrated the presence of the same amount of Pit-1 protein in rdw and normal rat pituitaries. The distribution of Pit-1-positive cells in the anterior pituitary was essentially the same in rdw and normal rats. It follows from these findings that the defective gene in the rdw rat is unrelated to the Pit-1 gene and the normal quantity of Pit-1 protein is insufficient to produce normal amounts of GH and PRL in the rdw pituitary. These and previous results suggest that the reduction in GH and PRL production in the rdw pituitary might be due to that in thyroid hormone production.

Amino Acid Sequence↗
PubMed1994

Fever of unknown origin: a review of 80 patients from the Shin'etsu area of Japan from 1986-1992.

In this survey involving 10 hospitals, we analyzed data on 80 Japanese patients from the Shin'etsu area (Nagano-ken and Niigata-ken) who were observed for fever of unknown origin (FUO). Our objectives were to identify the underlying causes and the relevant diagnostic methods. Fourteen of the patients died of the underlying illness. The cause of the FUO was infection in 43 patients, allergic or autoimmune disease in 13, neoplasm in 7, miscellaneous causes in 3, and undetermined in 14. FUO was self-limited in 13 patients and persistent in one patient. Methods successfully used to establish the final diagnosis in 66 patients were: evaluation of the clinical course or response to treatment in 16, serologic tests in 12, bacteriologic studies in 10, biopsy in 9, cytologic examination in 6, conventional radiology in 6, necropsy in 3, endoscopy in 2, and biochemical testing in 2.

Adolescent↗
PubMed1994

Molecular cloning of cDNA encoding the precursor for hamster hypothalamic growth hormone-releasing factor.

The structure of rat and mouse growth hormone-releasing factor (GRF) peptide and precursor shows considerable divergence from that of the human counterpart and also within rodents themselves. To study such structural divergence in another rodent, we cloned a cDNA encoding the GRF precursor from golden hamster. The hamster GRF (haGRF) cDNA clone had an open-reading frame that predicts a haGRF precursor protein with 107 amino acids. The haGRF precursor bore greater overall homology (82%) to the human than the same rodent homologue (58-64%) and contained two processing sites identical to the human sequence that would generate mature haGRF peptide. Furthermore, the haGRF peptide, like human but unlike rat or mouse GRF, consisted of 44 amino acids and also had greater homology to the human (89%) than the rodent sequence (64-75%), conserving a Tyr residue at the N-terminus and an amidated Leu residue at the C-terminus. Thus, both haGRF precursor and peptide are structurally more related to those of human than of other rodents, suggesting that rodent GRF precursor diverged from the human sequence at differential rates within the species.

Animals↗
PubMed1994

Physicochemical and biochemical characteristics of glycosylated recombinant human granulocyte colony stimulating factor (lenograstim).

Lenograstim, glycosylated recombinant human granulocyte colony stimulating factor (rHuG-CSF), is a glycoprotein that contains 4% O-glycosides with an approximate molecular weight of 20 kDa. The carbohydrate chain contributes to stabilisation of the protein by its ability to suppress polymerisation due to pH changes and to prevent denaturation at elevated temperatures. The carbohydrate chain also has an important role in maintaining biological stability in normal human serum. Furthermore, it is shown that glycosylated rHuG-CSF is more stable and resistant to degradation than non-glycosylated rG-CSF in vitro. In addition, we show the remarkable effects of combined therapy of lenograstim with an antibiotic in severe neutropenic infection models. These data demonstrate the possibility that such combination therapy may be applied to neutropenic patients with serious infections and that there is increased efficacy of antibiotic treatment when administered concurrently with lenograstim.

Adjuvants, Immunologic↗