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Y Kaneda

Publications and source records attributed to Y Kaneda.

At least 361 records · Page 20Linked to original sources
PubMed1994

A novel gene-transfer technique mediated by HVJ (Sendai virus), nuclear protein, and liposomes.

There have been many reports of in vivo direct gene transfer methods, but there are problems with these methods. Recently, we developed a novel, nontoxic and efficient in vivo direct gene transfer method mediated by Sendai virus (HVJ) and liposomes. In our system, foreign genes and nuclear proteins were encapsulated into the same liposomes, which were then treated with inactivated HVJ. The properties of our system are that HVJ enables foreign genes to be introduced directly into the cytoplasm by membrane fusion and that nuclear proteins transport the foreign genes rapidly into the nuclei. In the present study, we succeeded in introducing and expressing the functional gene for human insulin, and we observed the expression of this gene in mouse plasma and the reduction of plasma glucose. Our system will provide a new method of gene transfer aimed at postnatal gene therapy in various diseases.

Analysis of Variance↗
PubMed1994

[Modified Naclerio-Langer method under thoracoscopic procedure for pneumothorax due to rupture of a large bulla].

We managed a patient with pneumothorax caused by a large bulla by so called modified Naclerio-Langer method under thoracoscopic procedure. A 76-year-old female was hospitalized with recurrent left pneumothorax. Uncer the right lateral position, 3 Surgi ports were inserted through 5, 7, 9th intercostal space. There was a large bulla, 10 cm in diameter, with rupture at the ventral side. The bulla wall was partially resected and many bronchiolar openings were seen at the caudal side. Gregarious bronchiolar openings were resected with Endo GIA. Two residual openings were closed with 3-0 Dexon suture. The base of bulla was pleuralised by suturing the edges of the removed cyst with running suture of 3-0 Dexon. Because no air leaks was detected, a chest drain was inserted. Operation time was 215 minutes and she was discharged on 20th postoperative day. This procedure is usefull and not invasive.

Aged↗
PubMed1994

[Mediastinal lymph node dissection improves survival rate in N2(-) non-small cell lung cancer patients].

The effect of mediastinal lymph node dissection on survival rate in clinical N2(-) non-small cell lung cancer patients was assessed. N2(-) was evaluated by both pre-operative computed tomography studies and physical examination during surgery. Systematic mediastinal lymph node dissection, including nodes #1, 2, 3, 3a, 3p, 4, 7, 8 and 9 was performed in 27 patients (T1 or 2, N0 or 1, M0) since 1987 (dissected group). Survival rate and complications were compared with those in 21 patients treated before 1986, in whom mediastinal lymph nodes were not dissected (non-dissected group). 1, 3 and 5 year survival rates in the dissected group were 92.6%, 74.9%, and 74.9%, respectively, which were significantly higher than those in the non-dissected group (71.4%, 61.9% and 41.3%, respectively). Mediastinal lymph node metastases were detected histologically found in three patients, 11% of the dissected group. Respiratory complications showed a significantly higher incidence in the dissected group (p = 0.001). However these complications did not increase the mortality rate in that group. Among elderly patients over 70 years old (8 patients in the dissected group, 7 in the non-dissected group), survival rates were similar in the two groups. In conclusion, systematic mediastinal lymph node dissection significantly improved the survival rate in clinical N2(-) non-small cell lung cancer patients.

Aged↗
PubMed1993

Single intraluminal delivery of antisense cdc2 kinase and proliferating-cell nuclear antigen oligonucleotides results in chronic inhibition of neointimal hyperplasia.

To develop an effective strategy to prevent neointima formation after angioplasty injury, we have identified cell-cycle regulatory proteins as targets for inhibition by using antisense oligonucleotides (ODNs). We utilized an intraluminal molecular delivery method that employs the protein coat of a Sendai virus complexed with liposomes that enhances markedly the efficiency of ODNs uptake. First, we examined the effect of antisense cdc2 kinase and proliferating-cell nuclear antigen (PCNA) ODNs in vitro. Cotransfection of antisense cdc2 kinase and PCNA ODNs inhibited serum-stimulated vascular smooth muscle cell growth, whereas antisense cdc2 kinase ODNs alone or PCNA ODNs alone failed to show any inhibitory effect. Transfection of the combination of antisense cdc2 kinase and PCNA ODNs into balloon-injured arteries in vivo provided a marked decrease in cdc2 and PCNA mRNA expression as determined by reverse transcription-PCR, compared to sense controls. Antisense ODN treatment significantly inhibited the increase in DNA synthesis induced by balloon injury. Moreover, antisense ODN administration inhibited completely neointima formation at 2 weeks after angioplasty in an apparent dose-dependent manner. Moreover, the inhibitory effect of antisense ODN on neointima formation persisted up to 8 weeks after a single transfection. The present study documents that a single intraluminal molecular delivery of combined cdc2 kinase and PCNA antisense ODNs results in a sustained inhibition of neointima formation in the rat carotid balloon-injury model.

Animals↗
PubMed1993

Transfection of interleukin 2 gene into human melanoma cells augments cellular immune response.

A preclinical model was used to determine if transfection of the interleukin 2 (IL-2) gene into human melanoma cells would augment the response of autologous and allogeneic peripheral blood lymphocytes (PBLs) from melanoma patients. IL-2 gene was transfected into three human melanoma cell lines; secretion of IL-2 from stable transfected cells was confirmed by enzyme-linked immunosorbent assay. The PBL response to these melanoma cells was then examined in a mixed-lymphocyte tumor reaction using PBLs from eight melanoma patients. The PBL response to autologous (P < 0.01) or human leukocyte antigen A cross-reactive (P < 0.05) transfected melanoma cells was significantly higher than it was to nontransfected melanoma cells. These data suggest that IL-2 gene transfection may be an important strategic approach to enhancing specific immune responses induced by a polyvalent melanoma cell vaccine.

Base Sequence↗
PubMed1993

Construction of radiation-reduced hybrids and their use in mapping of microclones from chromosome 10p11.2-q11.2.

Radiation-reduced hybrids for mapping of DNA markers in the pericentromeric region of chromosome 10 were developed. A Chinese hamster/human somatic cell hybrid (762-8A) carrying chromosomes 10 and Y as the only human material were exposed to 40,000 rads of irradiation and then rescued by fusion with non-irradiated recipient Chinese hamster cells (GM459). Southern hybridization analyses revealed that 10 of 128 HAT-resistant clones contained human chromosomal fragments corresponding to at least one marker locus between FNRB (10p-11.2) and RBP3 (10q11.2). These hybrids were then used to map micro-dissection clones previously isolated and roughly mapped to this chromosomal region by fluorescence in situ hybridization (FISH). Two of the six microclones studied could be mapped to the proximity of the D10-S102 locus. These radiation hybrids are useful for the construction of refined genetic maps of the pericentromeric region of chromosome 10.

Animals↗
PubMed1993

Novel in vitro gene transfer method for study of local modulators in vascular smooth muscle cells.

Although many in vitro gene transfer methods already exist, such as calcium phosphate precipitation, electroporation, or cationic liposomes, these methods cause significant cell injury and cell death. The study of the biology of endogenous autocrine-paracrine vasoactive systems such as the renin-angiotensin system in vascular cells is limited by the lack of a suitable gene transfer method with high efficiency of transfection and expression that will permit cell biology studies. Recently, the Sendai virus (hemagglutinating virus of Japan, HVJ)-liposome-mediated gene transfer method has been shown to be an efficient and nontoxic method of gene transfer. In this study, we characterized the efficiency and suitability of the HVJ method for vascular biology research. Using SV40 T-antigen complementary DNA (cDNA), we initially compared the efficiency of the HVJ method and lipofection for transfection of cultured vascular smooth muscle cells (VSMCs). We observed that after 35 minutes of incubation, the HVJ method exhibited a 10-fold higher efficiency of transfection than lipofection. We used this method to study vascular angiotensin converting enzyme (ACE) expression in cultured VSMCs and cultured rat carotid arteries in vitro. The HVJ method of transfection of human ACE cDNA into VSMCs and COS cells was significantly more efficient than lipofection. Using this method, we demonstrated that transfection of ACE cDNA resulted in increased DNA synthesis, which was inhibited by the specific angiotensin II receptor antagonist DuP 753 (10(-6) M).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Hypertensive rats produced by in vivo introduction of the human renin gene.

We established an efficient and nontoxic in vivo gene transfer method mediated by the Sendai virus (hemagglutinating virus of Japan [HVJ]), liposomes, and nuclear protein. In this study, to produce a hypertensive model rat that is dependent on human renin, the human renin gene was introduced into adult rat liver by our efficient in vivo gene transfer method using HVJ and liposomes (HVJ-liposomes). The rats treated with HVJ-liposomes containing the human renin gene showed a significant elevation of blood pressure for 6 days compared with control rats, which received injections of HVJ-liposomes without the human renin gene. On day 5 after the transfer, human active renin as well as angiotensin II were found in the plasma of rats in which the human renin gene was introduced. Moreover, the blood pressure of these rats was significantly correlated with the plasma levels of human active renin and angiotensin II. To confirm that the elevated blood pressure was due to the expression of the human renin gene, we administered a newly developed specific human renin inhibitor, FK 906. The elevated blood pressure was normalized by the intravenous administration of this drug. These data indicate that this hypertensive rat was produced by the in vivo transfer of the human renin gene into rat liver and that the expressed human renin cleaved rat substrate (angiotensinogen). This hypertensive rat produced by in vivo gene transfer should be useful in further studies on hypertension.

Angiotensin II↗
PubMed1993

Novel and effective gene transfer technique for study of vascular renin angiotensin system.

Vascular renin angiotensin system (RAS) has been reported to exist in vascular wall. However, there is no direct evidence whether the vascular RAS per se can modulate growth of vascular smooth muscle cells (VSMC), because there is no suitable method to investigate the effect of endogenously produced vasoactive substances on growth of these cells. In this study, we transferred angiotensin-converting enzyme (ACE) and/or renin cDNAs into cultured VSMC using the efficient Sendai virus (hemagglutinating virus of Japan) liposome-mediated gene transfer method, to examine their relative roles in VSMC growth in vitro. Within 35 min or 6 h, the transfection of ACE cDNA into VSMC by hemagglutinating virus of Japan method resulted in a twofold higher ACE activity than control vector, whereas a cationic liposome (Lipofectin)-mediated method failed to show any effect. This in vitro system provided us with the opportunity to investigate the influence of endogenous vascular RAS on VSMC growth. Transfection of ACE or renin cDNA resulted in increased DNA and RNA synthesis, which was inhibited with the specific angiotensin II receptor antagonist (DuP 753: 10(-6) M). Angiotensin I added to ACE-transfected VSMC increased RNA synthesis in a dose-dependent manner. Cotransfection of renin and ACE cDNAs stimulated further RNA synthesis as compared to ACE or renin cDNA alone. These results showed that transfected components of RAS can modulate VSMC growth through the endogenous production of vascular angiotensin II, and that ACE as well as renin are rate limiting in determining the VSMC RAS activity. We conclude that the hemagglutinating virus of Japan liposome-mediated gene transfer technique provides a new and useful tool for study of endogenous vascular modulators such as vascular RAS.

Animals↗
PubMed1993

Glomerulosclerosis induced by in vivo transfection of transforming growth factor-beta or platelet-derived growth factor gene into the rat kidney.

Glomerulosclerosis, a final common lesion of various glomerular diseases, is characterized by mesangial cell proliferation and extracellular matrix (ECM) expansion. TGF-beta and PDGF are known to play a critical role in the regulation of ECM metabolism and mesenchymal cell proliferation, respectively. However, there is little evidence to demonstrate the direct role of each of these growth factors in the pathogenesis of glomerulosclerosis. Using an in vivo transfection technique, we could realize the selective overexpression of single growth factor in the kidney. The introduction of either TGF-beta or PDGF-B gene alone into the kidney induced glomerulosclerosis, although the patterns of action of these growth factors were different; TGF-beta affected ECM accumulation rather than cell proliferation and PDGF affected the latter rather than the former.

Animals↗
PubMed1993

The analysis of 40 kDa nuclear protein, p40, in interphase cells and mitotic cells.

We previously reported that the monoclonal antibody M108 recognized a 40 kDa protein both in the nucleus and the cytoplasm. This nuclear 40 kDa antigen was located in the nuclear envelope in interphase cells and in the perichromosomal region during mitosis. Now, we have analyzed this nuclear 40 kDa protein (p40) further, through morphological and biochemical approaches. At the beginning of mitosis, the perinuclear p40 detached from the nuclear envelope and moved to surround the condensing chromatin, while in the late stage of mitosis, the perichromosomal p40 moved back to the reassembled nuclear envelope. Most of the perichromosomal p40 on the metaphase chromosome was solubilized only by DNase I treatment, not by either high salt or detergent treatment. On the other hand, the perinuclear p40 was not solubilized by DNase1 alone, or high salt detergent alone. Sequential treatments with DNase I and high salt detergent were required to extract p40 in interphase nuclei. These results suggest that p40 was associated both with the nuclear envelope and chromatin DNA in interphase nuclei, while it bound only to chromatin DNA in mitosis.

Animals↗
PubMed1993

[Analysis of pathogenicity of Entamoeba histolytica by polymerase chain reaction].

DNA from five isolates of Entamoeba histolytica were examined for their pathogenicity by polymerase chain reaction. Three isolates SH-3,SH-6,SH-8 were isolated from patients with acute amoebic dysentery, whereas SH-5 and SH-7 were isolated from asymptomatic cyst passers. Gel electrophoresis of PCR products showed that primers P11, P12 for pathogenic strains could amplify genomic DNA extracted from SH-8, and primers P13, P14 for nonpathogenic strains could amplify genomic DNA extracted from SH-3,SH-5,SH-6 and SH-7. Furthermore, zymodeme analysis and the reactivity of McAb 4G6, which recognizes the 30 kDa antigen of pathogenic E. histolytica indicated that only SH-8 was pathogenic, while the others were nonpathogenic. The results of the genotypic analysis by PCR were in accord with the phenotypic properties. It is suggested that there are differences in genomic DNA between pathogenic and nonpathogenic strains. PCR is a highly sensitive and specific method for genomic DNA analysis of E. histolytica.

Animals↗
PubMed1993

[Chondrosarcoma of the trachea--report of a case].

A case of chondrosarcoma of the trachea in a 64-year-old man was reported. He was pointed out of benign tracheal tumor in September 1988. He was transferred to our hospital in September 1991 with respiratory distress attributable to asthma-like attack after bronchoscopic examination. Under right posterolateral thoracotomy, 1.7 cm length (3 rings) of the mediastinal trachea was resected and the trachea was reconstructed by end-to-end anastomosis with 3-0 Maxon interrupted sutures. The pathological diagnosis was a moderately well differentiated chondrosarcoma (grade II) of the trachea. Postoperative clinical course was uneventful without serious anastomotic complications. No local recurrence was seen 1 year after operation. It is necessary to follow meticulously the patients with chondrosarcoma of the trachea after surgery because of its low grade malignancy.

Chondrosarcoma↗
PubMed1992

Direct in vivo gene introduction into rat kidney.

We established a simple and highly efficient method for in vivo gene transfer using HVJ (Sendai virus) and liposomes. Plasmid DNA and high mobility group 1 (HMG1) protein were co-encapsulated in liposomes by agitation and sonication and were co-introduced into cells by HVJ-mediated membrane fusion. pACT SVT DNA, as a reporter gene, was introduced into the kidney of intact rats through a cannula in the renal artery, and SV40 large T antigen was detected by enzyme immunohistochemistry in glomerular cells 4 days after its introduction. This newly developed kidney-directed gene transfer method should be useful not only in basic research but also in potential gene therapeutics of renal diseases.

Actins↗
PubMed1992

A long synthetic peptide containing a nuclear localization signal and its flanking sequences of SV40 T-antigen directs the transport of IgM into the nucleus efficiently.

Synthetic short peptides containing only the nuclear localization signal (NLS) direct the transport of nonnuclear proteins into the nucleus. As a conjugate of the synthetic peptide with immunoglobulin M (IgM) did not enter the nucleus, there was believed to be a size limit for nuclear transport of NLS-conjugated proteins. However, we found that IgM conjugated with purified nucleoplasmin, a nuclear protein of Xenopus oocytes, rapidly accumulated in the nucleus. For direct comparison with the short peptide, we prepared a long peptide containing the NLS and its flanking sequences of SV40 large T-antigen and its mutated long peptide, in which possible phosphorylation sites located at the amino terminal of the NLS were changed to alanine. Kinetic experiments showed that wild-type long peptide-IgM conjugates were almost entirely taken up into the nucleus within 30 min after their injection, whereas almost 60 min was required for the mutated long peptide-IgM conjugates to enter the nucleus of all the cells examined, and there was no apparent accumulation of short peptide-IgM conjugates in the nucleus within 60 min. These results indicate that even when the kinetics of transport are affected by amino acid substitutions, the long peptide directs the transport of large molecules such as IgM into the nucleus.

Amino Acid Sequence↗