PubMed Health⌕ Search

Biomedical subjects

K Yoshimura

Publications and source records attributed to K Yoshimura.

At least 19 recordsLinked to original sources
PubMed1992

Tumor necrosis factor modulation of expression of the cystic fibrosis transmembrane conductance regulator gene.

Based on the knowledge that expression of the cystic fibrosis transmembrane conductance regulator (CFTR) gene can be modulated at the transcriptional level, and that the CFTR gene promoter contains sequences homologous to elements in other promoters that respond to tumor necrosis factor-alpha (TNF), we evaluated the hypothesis that TNF might modulate CFTR gene expression in epithelial cells. Studies with HT-29 cells, a colon epithelium-derived tumor cell line known to express the CFTR gene, demonstrated that TNF downregulated CFTR mRNA transcript levels in a dose- and time-dependent fashion. Interestingly, nuclear run-on analyses demonstrated that TNF did not affect the rate of transcription of CFTR gene, but exposure of the cells to TNF did modify the stability of CFTR mRNA transcripts, resulting in a mRNA half-life that was reduced to 65% of the resting level. These observations suggest that CFTR gene expression can be modulated by TNF, at least in part, at the posttranscriptional level.

Blotting, Northern↗
PubMed1992

Uveitis associated with human T-cell lymphotropic virus type I.

Seroepidemiologic, clinical, and virologic studies were performed to determine whether human T-cell lymphotropic virus type I was closely associated with uveitis in two hospitals. One hospital was in an endemic area of the virus (Miyakonojo, Miyazaki) and the other hospital was in a less endemic area (Kurume). In the endemic area, the seroprevalence of the virus in patients with uveitis without defined causes (35.4%, 62 of 175 patients) was significantly higher than that in patients with nonuveitic ocular diseases (16.1%, 42 of 261 patients), or in patients with uveitis with defined causes (10.3%, eight of 78 patients). The seroprevalence in younger patients (20 to 49 years of age) with uveitis without defined causes in the area was 44.8% (30 of 67 patients), whereas it was only 9.3% (ten of 107 patients) in the other two groups. A similar observation was recorded even in the less endemic area (Kurume). Because the seroprevalence of the virus in the general population is known to be low in younger patients and to increase with age, these findings were interpreted to indicate that the association of human T-cell lymphotropic virus type I with uveitis was significant. Most patients, particularly those aged 20 through 49 years, had an intermediate uveitis characterized by a moderate inflammation in the vitreous body accompanied by an iritis and retinal vasculitis. The ocular symptoms in the patients differed from those of other types of uveitis common in Japan (Behçet's disease, Vogt-Koyanagi-Harada's disease, and toxoplasmosis, for example).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

Expression of the cystic fibrosis transmembrane conductance regulator gene can be regulated by protein kinase C.

Epithelial cells utilize at least two types of apical Cl- channels, the cAMP-activated cystic fibrosis transmembrane conductance regulator (CFTR) and the Ca2+/calmodulin-dependent Cl- channel. While phorbal ester (PMA) activates only CFTR-dependent Cl- secretion and the Ca2+ ionophore A23187 only the Ca2+/calmodulin-dependent Cl- secretion, PMA and A23187 share the ability to down-regulate expression of the CFTR gene at the transcriptional level. Since both PMA and A23187 can activate protein kinases, we hypothesized that protein kinase pathways may be involved in the regulation of CFTR gene expression. Exposure of HT-29 human colon carcinoma cells to the protein kinase C activator SC9 down-regulated CFTR mRNA levels in a dose-dependent fashion, similar to that seen with PMA. The reduction in CFTR transcript levels by SC9 and PMA was blocked by H7, an inhibitor of protein kinases. In a similar fashion, the down-regulation of CFTR transcript levels by A23187 was blocked by H7 as well as staurosporine, another protein kinase inhibitor. Interestingly, both H7 and staurosporine themselves increased CFTR mRNA levels. Quantification of CFTR gene transcription rate showed a reduction by SC9 (similar to that with PMA and A23187) that was prevented by H7 and that H7 by itself increased CFTR transcription. Together, these observations suggest that protein kinase pathways, likely including protein kinase C, are involved in the regulation of CFTR gene expression, with activation or inhibition of protein kinase activity down-regulating or up-regulating CFTR gene expression, respectively.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1992

Expression of the human cystic fibrosis transmembrane conductance regulator gene in the mouse lung after in vivo intratracheal plasmid-mediated gene transfer.

As an approach to gene therapy for the respiratory manifestations of cystic fibrosis (CF), in vivo plasmid-mediated direct transfer of the normal CF transmembrane conductance regulator (CFTR) gene to the airway epithelium was investigated in mice. To evaluate the feasibility of this strategy, pRSVL, a plasmid composed of a firefly luciferase gene driven by the Rous sarcoma virus long terminal repeat (RSV-LTR), along with cationic liposomes was instilled into the trachea of C57BI/6NCR mice. With administration of 200-400 micrograms plasmid DNA, luciferase expression could be detected in the mouse lung homogenates for at least 4 wk. With this background, a CFTR expression plasmid vector (pRSVCFTR) constructed by replacing the luciferase cDNA from pRSVL with the normal human CFTR cDNA was evaluated in vivo in mice. Intratracheal instillation of pRSVCFTR with cationic liposomes followed by analysis of mouse lung RNA by polymerase chain reaction amplification (after conversion of mRNA to cDNA) using a RSV-LTR specific sense primer and a human CFTR-specific antisense primer demonstrated human CFTR mRNA transcripts from one day to 4 wk after instillation. Further, in vivo evaluation of beta-galactosidase activity after intratracheal administration of an E. coli lacZ gene expression plasmid vector directed by the cytomegalovirus promoter (pCMV beta) demonstrated that the airway epithelium was the major target of transfer and expression of the exogenous gene. These observations demonstrate successful plasmid-mediated gene transfer to the airway epithelium in vivo. This strategy may be feasible as a form of gene therapy to prevent the pulmonary manifestations of CF.

Animals↗
PubMed1992

Transcriptional and posttranscriptional modulation of human neutrophil elastase gene expression.

Human neutrophil elastase (NE), a 29-Kd potent serine protease stored in azurophilic granules of mature neutrophils, is coded for by the NE gene, a single copy gene with 5 exons spanning a 6-kb segment of chromosome 11 at q14. With the knowledge that the NE gene expression is limited to early myeloid cell differentiation, mechanisms modulating expression of the NE gene were evaluated in the HL-60 promyelocytic leukemia cell line, a model of early bone marrow precursor cells. Consistent with the presence of NE messenger RNA (mRNA) transcripts in undifferentiated HL-60 cells, nuclear transcription run-on analyses showed that HL-60 cells actively transcribed the NE gene. However, the transcription rate of the NE gene was relatively low, only 40% of the myeloperoxidase gene, a gene expressed in parallel with NE. When induced toward the mononuclear phagocytic lineage with phorbol 12-myristate 13-acetate (PMA), HL-60 cells exhibited marked suppression of NE gene transcription, declining to 17% of the resting rate within 2 days. Induction toward mononuclear phagocytic lineage differentiation caused no change in NE mRNA transcript half-life (T1/2), but mRNA levels decreased markedly over time, with levels undetectable 1.5 days after PMA stimulation. In contrast, when induced toward the myelocytic lineage with dimethyl sulfoxide, the rate of NE gene transcription increased 1.9-fold within 5 days. Interestingly, the mRNA transcript levels increased 2.5-fold by 5 days despite the fact that induction toward myelocytic lineage differentiation was accompanied by a marked reduction of NE mRNA transcript T1/2. Together, these observations suggest that the NE gene expression during bone marrow differentiation is modulated mainly at the transcriptional level, with some posttranscriptional modulation contributing, particularly during myelocytic lineage differentiation.

Blotting, Northern↗
PubMed1992

Interaction between the calcium and cyclic AMP messenger systems in perifused rat parotid acinar cells. Possible mechanism for potentiation of amylase secretion.

Potentiation of amylase secretion induced by a combination of isoproterenol and carbamylcholine was examined in perifused rat parotid acinar cells. The time course of changes in the augmented amylase secretion induced by isoproterenol plus carbamylcholine was similar to that induced by carbamylcholine alone, but not to that caused by isoproterenol. Concentration-response analysis showed that isoproterenol increased the apparent affinity for carbamylcholine to stimulate amylase secretion with the maximum effect attained by isoproterenol plus carbamylcholine being higher than that attained by isoproterenol or carbamylcholine. 8-Bromo cyclic AMP, forskolin and 3-isobutyl-1-methylxanthine mimicked the effect of isoproterenol. Calcium ionophores (A23187 and ionomycin), but not phorbol 12,13-dibutyrate, mimicked the effect of carbamylcholine. Chelation of intracellular free calcium with 1,2-bis-[2-aminophenoxyl]-ethane-N,N,N',N'-tetraacetic acid, but not that of extracellular calcium with [ethylenebis(oxyethylenenitrile)]-tetraacetic acid (EGTA), abolished the potentiation. Calmodulin antagonists inhibited amylase secretion induced by isoproterenol plus carbamylcholine or carbamylcholine alone, but not that induced by isoproterenol alone. These results suggest that the potentiation is mainly, if not completely, caused by a coordinated interaction between the cyclic AMP system and the Ca2+ system at a step distal to second messenger generation, probably via a cyclic AMP-induced increase in the sensitivity of the Ca2+ response element to calcium.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1992

In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium.

Direct transfer of the normal cystic fibrosis (CF) transmembrane conductance regulator (CFTR) gene to airway epithelium was evaluated using a replication-deficient recombinant adenovirus (Ad) vector containing normal human CFTR cDNA (Ad-CFTR). In vitro Ad-CFTR-infected CFPAC-1 CF epithelial cells expressed human CFTR mRNA and protein and demonstrated correction of defective cAMP-mediated Cl- permeability. Two days after in vivo intratracheal introduction of Ad-CFTR in cotton rats, in situ analysis demonstrated human CFTR gene expression in lung epithelium. PCR amplification of reverse transcribed lung RNA demonstrated human CFTR transcripts derived from Ad-CFTR, and Northern analysis of lung RNA revealed human CFTR transcripts for up to 6 weeks. Human CFTR protein was detected in epithelial cells using anti-human CFTR antibody 11-14 days after infection. While the safety and effectiveness remain to be demonstrated, these observations suggest the feasibility of in vivo CFTR gene transfer as therapy for the pulmonary manifestations of CF.

Adenoviruses, Human↗
PubMed1992

Role of prostaglandin E2 in cholinergic-mediated glycoprotein synthesis in canine antrum.

We studied the mechanism of cholinergic stimulation of mucin synthesis in canine antral explants, including the role of PGE2 as an intermediate messenger. Isolated antral mucosa was incubated with 10(-5) M carbachol (Cb), 10(-5) M indomethacin (IND), 10(-5) M pirenzepine (PZ), 10(-5) M Cb + 10(-5) M PZ, 10(-5) M Cb + 10(-5) M IND, and 10(-5) M IND + PGE2 (10(-8), 10(-7) and 10(-6) M) in the presence or absence of [3H]glucosamine. After 24 hr, total glycoprotein synthesis was quantitated by Sepharose-4B chromatography and by 10% TCA/1% PTA precipitation with lipid extraction. PGE2 released into the media was measured by radioimmunoassay (RIA). Cb significantly increased total glycoprotein synthesis and produced a significant increase in PGE2 release. The increase in glycoprotein synthesis and the release of PGE2 was blocked by the addition of muscarinic antagonist PZ. The addition of IND significantly inhibited glycoprotein synthesis and almost entirely suppressed PGE2 secretion. IND also inhibited the effect of Cb on glycoprotein synthesis and PGE2 release. Moreover, PGE2 (10(-6) and 10(-7) M) significantly increased the glycoprotein synthesis in the canine stomach. This suggests the coordinate participation of PGE2-releasing cell population in modulation of glycoprotein synthesis in gastric mucosa.

Animals↗
PubMed1992

Effect of sucralfate on components of mucosal barrier produced by cultured canine epithelial cells in vitro.

The mucous gel maintains a neutral microclimate at the epithelial cell surface, which may play a role in both the prevention of gastroduodenal injury and the provision of an environment essential for epithelial restitution and regeneration after injury. Enhancement of the components of the mucous barrier by sucralfate may explain its therapeutic efficacy for upper gastrointestinal tract protection, repair, and healing. We studied the effect of sucralfate and its major soluble component, sucrose octasulfate (SOS), on the synthesis and release of gastric mucin and surface active phospholipid, utilizing an isolated canine gastric mucous cells in culture. We correlated these results with the effect of the agents on mucin synthesis and secretion utilizing explants of canine fundus in vitro. Sucralfate and SOS significantly stimulated phospholipid secretion by isolated canine mucous cells in culture (123% and 112% of control, respectively). Indomethacin pretreatment significantly inhibited the effect of sucralfate, but not SOS, on the stimulation of phospholipid release. Administration of either sucralfate or SOS to the isolated canine mucous cells had no effect upon mucin synthesis or secretion using a sensitive immunoassay. Sucralfate and SOS did not stimulate mucin release in the canine explants; sucralfate significantly stimulated the synthesis of mucin, but only to 108% of that observed in untreated explants. No increase in PGE2 release was observed after sucralfate or SOS exposure to the isolated canine mucous cells. Our results suggest sucralfate affects the mucous barrier largely in a qualitative manner. No increase in mucin secretion or major effect on synthesis was noted, although a significant increase in surface active phospholipid release was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

The phase of sperm flagellar beating is not conserved over a brief imposed interruption.

We have studied the phase component of flagellar beating by holding the head of a sea urchin sperm in the tip of a sinusoidally vibrating micropipet and then abruptly displacing the pipet laterally at a speed of 2.5 microns/ms for various durations. This rapid displacement of the pipet delayed the initiation of the next bend for as long as the displacement continued, up to a duration of 1 beat cycle, corresponding to a delay of 0.5 beat cycle. At the end of this displacement, the movement of the pipet was stopped completely without resumption of the initial vibration. Analysis of the flagellar waveform showed that immediately when the pipet was stopped, the flagellum started to beat by spontaneously initiating the bend that had been delayed. The flagellum then continued steady-state beating, with normal waveform and a new phase that was independent of the original phase of beating. These data suggest that the information on the phase of beating is located only at the basal end of the flagellum, and not in oscillators distributed along the axoneme. After this information has been lost, the flagellum can resume beating at any arbitrary phase relative to its original phase.

Animals↗
PubMed1992

Millardia meltada, a new host for Acanthocheilonema viteae and a simple technique for separation of microfilariae from peripheral blood.

Millardia meltada were infected with Acanthocheilonema viteae and examined for their susceptibility. The morbidity of infected M. meltada was low compared with that of jirds. On day 47 post-infection (p.i.), 13 of 14 M. meltada developed microfilaremia. Male M. meltada then showed gradually increasing microfilaremia with a peak level of 7000 per 30 microliters blood at week 20 p.i., which was much higher than that (3000) of male jirds. In contrast, microfilarial densities of female M. meltada were markedly low with a peak level of 200 during weeks 10-12 p.i. A simple centrifugation technique with Lympholyte-M was devised for microfilarial separation from the peripheral blood of infected M. meltada and yielded approximately 17 x 10(5) viable microfilariae from 1 ml of blood. This method also makes it possible to collect microfilariae from the same individuals repeatedly. M. meltada, coupled with this microfilarial separation technique, serves as a useful animal model for microfilarial studies of A. viteae.

Animals↗
PubMed1992

The influence of Mesocestoides corti on subsequent Angiostrongylus cantonensis infections in mice.

The influence of Mesocestoides corti on subsequent Angiostrongylus cantonensis infection in mice (C57BL/6 and BALB/c) was assessed. Both strains of mice infected with M. corti demonstrated a peak blood eosinophilia at around 3 weeks post-infection (p.i.). C57BL/6 and BALB/c mice primarily infected with M. corti were given A. cantonensis infection 18 days later, but pre-existing M. corti infection did not affect the recovery of intracranial worms of A. cantonensis at day 21 p.i. BALB/c mice with mixed parasite infections showed low morbidity and mortality as compared with mice singly infected with A. cantonensis and some mice demonstrated a pulmonary migration of intracranial worms. In C57BL/6 mice, intracranial worms were killed and thus all mice survived. C57BL/6 mice with mixed parasite infections failed to resist A. cantonensis reinfection. The blastogenic responses of spleen cells against A. cantonensis antigen were lower in BALB/c than in C57BL/6 mice and mixed parasite infections also resulted in less blastogenic responses against both concanavalin A and A. cantonensis antigen than monoinfection. The recovery of M. corti biomass was significantly higher in mice with mixed parasite infections than mice with monoinfection with M. corti. These data suggest a distinct difference in response to A. cantonensis infection between C57BL/6 and BALB/c mice, and the induction of immunosuppression in both mouse strains following M. corti infection. Blood eosinophilia provoked by M. corti infection is not directly associated with the killing of worms in subsequent A. cantonensis infection.

Angiostrongylus cantonensis↗
PubMed1992

Restoration of the defective natural defence of beige mice against tissue-migrating larvae of Strongyloides ratti by transfer with normal peritoneal cells.

The effects of cell transfer on the defective natural defences of beige (bgj/bgj) mice against Strongyloides ratti were studied by assessing recovery of tissue-migrating larvae from head and lung. Transfer of peritoneal resident cells from normal bgj/+ mice restored the defective natural defence of beige mice. The non-adherent population of normal peritoneal cells did not have the restorative capacity. Macrophages may be important to the natural defence against S. ratti.

Animals↗
PubMed1992

Effects of polyoxyethylene (20) sorbitan monooleate on the acute toxicity of linear alkylbenzenesulfonate (C12LAS) to fish.

Effects of polyoxyethylene (20) sorbitan monooleate (SMOE20) on the acute toxicity of linear alkylbenzenesulfonate (C12LAS) to fish were investigated in red killifish (Orizias latipes) and carp (Cyprinus carpio). By adding polyoxyethylene sorbitan ester to C12LAS solution, the acute toxicity of C12LAS decreased. As a decreasing toxic effect of these nonionic surfactants, the depression of gill damage was histopathologically observed. No significant hematological and blood biochemical differences were observed between two tests using a single C12LAS solution and a mixed solution of C12LAS/SMOE20. The addition of SMOE20 to C12LAS decreased the incorporation of C12LAS into blood and each organ except spleen and gall bladder as well as the adsorption of C12LAS to the gill. The farther inward the phenyl position in the alkyl side chain, the lower the adsorption of C12LAS to gill. Since the acute toxicity of LAS phenyl isomers to fish was known to decrease according to the order of adsorption, these results suggest that the decreasing toxic effect of SMOE20 on the acute toxicity of C12LAS is due to depression of more toxic C12LAS adsorption on the gill.

Adsorption↗
PubMed1992

Postsurgical inflammation after phacoemulsification and extracapsular extraction with soft or conventional intraocular lens implantation.

A one-year prospective study was conducted in 120 patients to assess the time course of changes in intraocular inflammation after three cataract surgery procedures: planned extracapsular extraction with poly(methyl methacrylate) (PMMA) intraocular lens (IOL) implantation (11 mm incision group), phacoemulsification with PMMA IOL implantation (7 mm incision group), and phacoemulsification with foldable silicone single-piece IOL implantation (4 mm incision group). Each group was carefully matched for patients' ophthalmologic and systemic backgrounds. Patients with hard nuclei were excluded. The degree of inflammation was evaluated by quantitating aqueous flare intensity and cell count with the laser flare-cell meter. In the early postoperative period, both aqueous flare intensity and cell count were highest in the 11 mm incision group followed, in decreasing order, by the 7 mm and 4 mm incision groups. Significant between-group differences were observed at one, two, and seven postoperative days for flare and one day through one week for cells. Both parameters in each group decreased to a similar level one month after surgery, but flare intensity in all groups remained significantly higher than that of age-matched normal controls up to six months postoperatively.

Aged↗
PubMed1992

Protease resistant interleukin-3 stimulating components in excretory and secretory products from adult worms of Strongyloides ratti.

Excretory and secretory (ES) products collected from adult worms of Strongyloides ratti stimulated interleukin-3 (IL-3) production with mesenteric lymph node cells from infected C57BL/6 mice, but not with normal mesenteric lymph node cells. The IL-3 stimulating components were not major IgG binding antigens. Activity of the IL-3 stimulating components was stable by treatment with protease, although reduced by heating in boiling water.

Animals↗
PubMed1992

A case of adult T cell leukemia with bullae in the palmoplantar regions followed by a crisis.

A 48-year-old Japanese female who had had chronic ATL for 4 years suddenly developed vesicles on the palms and soles. Histologically, these bullae were specific lesions of ATL. After a tendency toward improvement, a crisis appeared with increases in the following: peripheral white blood cells, atypical lymphocytes, CD25 positive cells, serum LDH, and soluble IL-2R. Palmoplantar bullae, a rather rare finding, may be indicative of a following crisis.

Female↗