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Biomedical subjects

Y Kaneda

Publications and source records attributed to Y Kaneda.

At least 487 records · Page 27Linked to original sources
PubMedDate not supplied

EBV replicon vector system enhances transgene expression in vivo: applications to cancer gene therapy.

BACKGROUND: A major limitation of current cancer gene therapies is low and transient expression of the therapeutic gene. For long-term expression of transgenes in vivo, an Epstein-Barr virus (EBV) replicon vector has been developed. The present study examines the effect of the EBV replicon vector system and its application to a suicide gene therapy for melanoma in mice. METHODS: An EBV replicon vector system, pEBc, consisting of EBV nuclear antigen-1 (EBNA-1) and the origin of latent viral DNA replication, oriP, was used to express either the luciferase gene or the herpes simplex virus (HSV) thymidine kinase (TK) gene. The expression vector was introduced in vivo into melanoma tumor masses in mice by means of HVJ-cationic liposomes. The time-course of gene expression and the anticancer effect of the EBV replicon vector were investigated in comparison with pcLuc, which lacks the EBV components. RESULTS: Luciferase expression was sustained in both cultured cells and melanoma masses by pEBc but not by pcLuc. The luciferase expression level in melanoma masses was higher by pEBcLuc than by pcLuc, although Southern blot analysis showed the number of copies of pEBcLuc retained in the melanoma masses to be fewer than that of pcLuc. The effectiveness of EBV replicon vector on suicide gene therapy of melanoma in mice was also demonstrated. CONCLUSION: The EBV replicon vector appears useful for cancer gene therapy. Analysis of the transgene in tumors suggests that the EBV replicon system may be responsible for efficient transcription but not retention of the transgene.

Animals↗
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Characterization of local inflammatory response in an isolated lung perfusion model.

BACKGROUND: Current phase I trials of isolated lung perfusion for treatment of pulmonary metastases have an arbitrarily determined length of perfusion. Our objective was to examine the temporal course of the local and distant inflammatory response as a function of the length of perfusion (ischemia) and subsequent reperfusion in an equivalent animal model. METHODS: Sixty male Fischer 344 rats were randomized into four groups (n = 15). Each group underwent left isolated lung perfusion with buffered Hespan for 10, 30, 60, or 90 minutes. Subsequently, two subgroups of five animals within each group were allowed to reperfuse for 1 or 3 hours, respectively. Non-perfused right lung was used as control. At each time point, lung specimens were assayed for TNF-alpha by ELISA and histologic sections were examined. RESULTS: There was no significant difference between the left and right lung tissue levels of TNF-alpha at the termination of the ischemic period. However, on reperfusion, the left lung TNF-alpha levels increased significantly above the ischemia baseline in all groups, with a greater magnitude of rise in the groups with 60 and 90 minutes of preceding ischemia (12,757 +/- 1985 vs. 3524 +/- 494 pg/g, and 16,914 +/- 1657 vs. 6530 +/- 1104 pg/g, respectively; p < 0.05). There was no significant elevation in tissue levels of TNF-alpha in the right lung. Histologic changes consistent with early pulmonary edema were first detected at 12 hours following onset of reperfusion. CONCLUSIONS: Reperfusion following prolonged pulmonary ischemia during isolated lung perfusion results in a significant elevation of local tissue levels of TNF-alpha and may render the perfused lung vulnerable to the adverse effects of the inflammatory cascade.

Animals↗
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Development of a novel fusogenic viral liposome system (HVJ-liposomes) and its applications to the treatment of acquired diseases.

Toward human gene therapy and gene analysis in vivo, a novel hybrid vector based on liposome has been developed for more efficient gene delivery and gene expression. The liposome was decorated with HVJ (Sendal virus) envelope fusion proteins to introduce DNA directly into the cytoplasm, and contained DNA and DNA-binding nuclear protein to enhance expression of the gene. Recently, several types of HVJ-liposomes were developed by altering the lipid components of the liposomes. HVJ-cationic liposomes increased gene delivery 100-800 times more efficiently in vitro than the conventional HVJ-anionic liposomes. HVJ-cationic liposomes were also more useful for gene expression in restricted portions of organs and for gene therapy of disseminated cancers. It was further discovered that the use of anionic liposomes with a virus-mimicking lipid composition (HVJ-AVE liposomes) increased transfection efficiency by several fold in vivo, especially in liver and muscle. By coupling the Epstein-Barr (EB) virus replicon apparatus to HVJ-liposomes, transgene expression was sustained in vitro and in vivo. Most animal organs were found to be suitable targets for the fusigenic-viral liposome system, and numerous gene therapy strategies using this system were successful in animals.

Gene Expression↗
PubMedDate not supplied

Minute fragment observed in a bovine pedigree with Robertsonian translocation.

A minute fragment was found in the metaphase plates obtained from one Holstein bull carrying Robertsonian translocation. The pedigree investigation indicated that three out of 34 offspring had the same minute fragment as their sire. Fluorescence in situ hybridization (FISH) studies with bovine satellite DNA revealed that the minute fragment was hybridized with this DNA and was part of the bovine centromere. We propose that the minute fragment is the product of centric fusion and is transmitted to offspring without being lost in the subsequent cell divisions.

Animals↗
PubMedDate not supplied

Reactivity of monoclonal antibodies to species-specific antigens of Entamoeba histolytica.

Twenty monoclonal antibodies were produced against trophozoites of Entamoeba histolytica strains HK-9 and HM-1: IMSS. When reactivity to various enteric protozoa was examined by an indirect fluorescence antibody test, 15 of the monoclonal antibodies were strongly reactive with E. histolytica trophozoites. Species-specific antigens recognized by these monoclonal antibodies were located on the plasma membrane, nucleus, cytoplasm, and cytoskeletal structures of the trophozoites. Two of the remaining five monoclonals reacted strongly with trophozoites of the E. histolytica-like Laredo strain. The determinant antigen was located in the cytoplasm. The three remaining monoclonal antibodies were found to recognize cross-reactive antigens between E. histolytica and E. histolytica-like Laredo, E. hartmanni, E. coli, Dientamoeba fragilis, Giardia lamblia, and Trichomonas hominis. These three antibodies were also reactive with T. vaginalis and mammalian cells such as HeLa cells. Thus, the combined use of monoclonal antibodies seems capable of distinguishing E. histolytica and/or E. histolytica-like Laredo from other enteric protozoa.

Animals↗
PubMedDate not supplied

The HVJ liposome method.

Recent advancement of gene technology allows us a practical approach to gene therapy. Among various in vivo gene transfer techniques available, the HVJ liposome method is an efficient procedure which could target the glomerular cells. Using this method, HVJ-mediated cell fusion activity enables us to introduce genetic materials directly into the cytosol without degradation. In addition, cointroduction of non-histone nuclear protein, high-mobility group (HMG-1), efficiently facilitates migration of foreign DNA to the nucleus. Although there still exist some limitations, the HVJ liposome method may be applicable to the treatment of glomerular diseases as well as to analysis of the molecular aspects of renal pathophysiology.

Animals↗
PubMedDate not supplied

22-Oxacalcitriol upregulates p21(WAF1/Cip1) in human parathyroid glands. A preliminary report.

In the era of 22-oxacalcitriol (OCT), newly synthesized 1 alpha,25-dihydroxyvitamin D(3) analogue, against secondary hyperparathyroidism, the indications of parathyroidectomy (PTx) has been restricted. Recent investigations on animal models have revealed the inhibitory effects on PTH secretion after OCT treatment, whereas there has been no evidence about human parathyroid glands. A 38-year-old man with a 19-year history of hemodialysis was performed PTx after the failure of OCT treatment. Expressions of proliferative nuclear cell antigen (PCNA), calcium-sensing receptor (CaSR), vitamin D receptor (VDR), p53 and p21(WAF1/Cip1) were analyzed by Western blotting and immunohistochemistry on resected parathyroid glands. We confirmed up-regulations of CaSR and VDR, which contribute the reduction of serum PTH, by OCT treatment. Concomitant up-regulation of p21(WAF1/Cip1) but not p53, especially in nodular hyperplasia, can be considered to induce cell cycle arrest of the parathyroid cells, but not cytocidal effect of OCT.

Adult↗
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In vitro and in vivo antibacterial activity and pharmacokinetics of SC-002 and its derivative, SC-004: new oral cephalosporins.

SC-002 is a novel oral cephalosporin possessing a unique thiadiazolylethenyl moiety at the 3 position. In the present study, it was the most active against gram-positive bacteria among oral cephalosporins such as cefdinir (CFDN), cefpodoxime, cefditoren and cefaclor (CCL). It was equal to or 16 times more active than CFDN against standard and clinical strains. In particular, against clinical isolates of Morganella morganii and Haemophilus influenzae, SC-002 was 8-64 times more active than CFDN. The antibacterial activity of SC-002 against some beta-lactam-resistant strains was superior to that of CFDN. The in vivo antibacterial activity of SC-004, a pivaloyloxymethyl ester of SC-002, was 1.2-8 times more protective against systemic infections due to Streptococcus pneumoniae, Escherichia coli and Klebsiella pneumoniae than that of CFDN. The therapeutic effects of SC-004 on experimental respiratory tract infections caused by S. pneumoniae or H. influenzae were superior to those of CFDN and CCL. SC-004 showed higher and longer-lasting blood levels and higher urinary excretion in pharmacokinetics in mice.

Administration, Oral↗
PubMedDate not supplied

Experimental study on external tibial rotation of the knee.

Using biplanar roentgenographic photogrammetry, we investigated posterolateral rotatory instability of the knee joint both before and after sectioning of posterolateral structures, the posterior cruciate ligament, and the lateral collateral ligament. Fifteen fresh-frozen cadaveric knees were used. Compared with the intact state, sectioning of the posterior cruciate ligament alone did not increase the amount of external tibial rotation, but the axis of external tibial rotation shifted when the anterolateral bundle of the posterior cruciate ligament was cut. When the posterior cruciate ligament was cut after sectioning of the posterolateral structures and the lateral collateral ligament, external tibial rotation increased and the axis of external rotation shifted. The results demonstrated that sectioning of the anterolateral bundle of the posterior cruciate ligament is associated with a change in the location of the axis of tibial rotation. Therefore an isolated posterior cruciate ligament injury can alter the kinematics of the knee joint by changing the axis of external tibial rotation. The present results also demonstrate that the posterior cruciate ligament serves as a kind of secondary restraint to posterolateral rotatory instability in knees with injured posterolateral structures. Helical motion analysis using biplanar roentgenographic photogrammetry is a useful method for evaluating knee kinematics.

Aged↗
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Epidemiological survey of Trypanosoma cruzi infection in north-eastern Brazil using different diagnostic methods.

A survey of the prevalence of Trypanosoma cruzi infection was carried out in Oitis, a small community in the State of Piaui, Brazil. Two hundred and sixty five individuals were screened by microscopic examination, hemoculture, indirect immunofluorescence (IFA), enzyme-linked immunosorbent assay (ELISA), and competitive enzyme-linked immunosorbent assay (C-ELISA) using the monoclonal antibody TCF87 against to a 25kd T. cruzi antigen. Seropositivity was 14.3% by the IFA test, 14.7% by ELISA, and 13.2% by C-ELISA. The C-ELISA using the TCF87 monoclonal antibody seems to be applicable in serodiagnosis of Chagas' disease.

Adolescent↗
PubMedDate not supplied

Allogeneic class I major histocompatibility complex gene transfer in murine neuroblastoma in vivo.

In vivo intratumoral gene transfer of allogeneic class I major histocompatibility complex (MHC) genes augments the immune response against weak tumor antigens. In this study, mice inoculated with the allogeneic MHC molecule (H-2Kb), had transduced-murine neuroblastoma C1300S3 cells showed prolonged survival relative to non-transduced or neo transduced tumors (p < 0.005). Interestingly, direct in vivo gene transfer of H-2Kb plasmid DNA complexed with HVJ-liposomes into S3 tumors was highly efficient, resulting in transduction of 8% of the interstitial cells within the tumor but rarely within tumor cells. Regression of established tumors and prolonged survival occurred in 50% of mice injected with H-2Kb, in contrast to no tumor regression in mice receiving control plasmid (p < 0.005). This study concludes that interstitial cells could serve as an important target of intratumoral gene transfer, and further that HVJ-liposome complexes could be a vehicle for in vivo gene transfer.

Animals↗
PubMedDate not supplied

Digitonin enhances the anti-tumor effect of cisplatin against methylcholanthrene-induced rat sarcoma cells in vitro.

BACKGROUND: This study was designed to evaluate cellular uptake and cytotoxicity of cisplatin in methylcholanthrene (MCA)-induced rat sarcoma cells when used in combination with a detergent, digitonin. MATERIALS AND METHODS: In the cellular intake study, after MCA sarcoma cells (10(7)) were treated with cisplatin alone (50 micrograms/ml) and with cisplatin (50 micrograms/ml) in combination with digitonin at 20 microM and 50 microM, the cells were washed twice with PBS and the platinum levels were measured by flameless atomic spectrometry. For the anti-tumor effect MCA sarcoma cells (10(3)) were seeded in cell culture dishes and loaded for 10 minutes with PBS, digitonin (5 microM), cisplatin (5 micrograms/ml) or a combination of cisplatin (5 micrograms/ml) and digitonin (5 microM). The cells were then washed and incubated for 72 hours. Bromodeoxyuridine uptake was measured with an ELISA system for determining viable cells counts. RESULTS: Cell platinum levels were significantly elevated in proportion to the increase of digitonin (p < 0.0001). The number of viable cells was significantly decreased with, the combined cisplatin (5 micrograms/ml)--digitonin (5 microM) treatment (p < 0.0001 CONCLUSION: Digitonin enhances the antitumor effect of cisplatin against methylcholanthrene-induced rat sarcoma in vitro.

Animals↗