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Results 701 - 750 of 1868
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Dailey L A - - 2004
This study examined the effect of nebulization on the cellular uptake and transfection efficiency of polyplexes from four polyethylenimine (PEI) modifications: branched 25 kDa PEI (bPEI), linear 22 kDa PEI (linPEI), pegylated PEI (pegPEI) and biodegradable PEI (bioPEI). Polyplexes were aerosolized with air-jet and ultrasonic nebulizers. The aerosol was collected ...
Farson Deborah - - 2004
BACKGROUND: One of the major limitations to the use of adeno-associated virus (AAV) vectors for gene therapy has been the difficulty in producing enough vector to supply a clinical trial. More than 20 000 roller bottles may be required to generate AAV by the traditional transient transfection process to treat ...
Jin Ying - - 2004
1. This study describes the use of an oligodendroglial cell line (158N) to study the protective effects of X-chromosome-linked inhibitor of apoptosis (XIAP) overexpression. 2. 158N cells were transiently transfected with either pCMV-Myc-XIAP or control pCMV-Myc vector. At 48 h post-transfection, immunoblotting and immunocytochemical staining showed robust myc-XIAP overexpression in ...
Henderson Gail - - 2004
Infection of cattle or bovine cells with bovine herpesvirus 1 (BHV-1) leads to increased apoptosis. Previous studies indicated that BHV-1 infected cell protein 0 (bICP0), the major transcriptional regulatory protein of BHV-1, is toxic in transiently transfected cells. Point mutations within the zinc RING finger of bICP0 reduced toxicity and ...
Chen Qinzhong - - 2004
PURPOSE: We have previously confirmed that cyclooxygenase-2 (COX-2) is expressed in a human renal cell carcinoma (RCC) cell line and it has an important role in cell tumorigenesis and angiogenesis. In the current study we evaluated the impact on cell adhesion and tumor invasiveness in human RCC cell lines by ...
Smales C M - - 2004
We have employed an inverse engineering strategy based on quantitative proteome analysis to identify changes in intracellular protein abundance that correlate with increased specific recombinant monoclonal antibody production (qMab) by engineered murine myeloma (NS0) cells. Four homogeneous NS0 cell lines differing in qMab were isolated from a pool of primary ...
Abdul-Wahid A - - 2004
Cyst wall proteins 1 and 2 (CWP1 and CWP2) are major constituents of the giardial cyst wall and are expressed with similar kinetics by encysting trophozoites. In the present study, we were interested to determine if the expression of giardial CWPs as heterologous proteins in a higher eukaryotic cell would ...
Endesfelder Stefanie - - 2005
Increased levels of the cyclin-dependent kinase inhibitor p21 associated with decreased myoblast proliferation may be involved in the dystrophic process in Duchenne muscular dystrophy (DMD). Therefore we are interested to improve the proliferation of primary myoblasts of DMD patients by a reduction in p21 using either antisense oligonucleotides (ASO) or ...
Zhou Qiong - - 2004
Expression of polo-like kinase 1 (Plk1) is elevated in lung cancer and has been proposed as having prognostic value and related to resistance to chemotherapy and radiation. In addition, Plk1 has several functions in mitotic progression. In this study, the authors investigated the effect of Plk1 depletion on cell cycle ...
Hampson Lynne - - 2004
Previous work has indicated that the PDZ domain Tax interacting protein 1 (Tip-1) is a target of the HTLV1 Tax protein and is a potential RhoA effector. We have used the yeast two-hybrid system to show that Tip-1 also interacts with the HPV16 E6 protein. This interaction was confirmed by ...
Mason J I - - 2004
3Beta-hydroxysteroid dehydrogenase (3beta-HSD) activity is essential for the synthesis of all classes of steroid hormones, converting various delta5-3beta-hydroxysteroids into hormonally active delta4-3-ketosteroids in NAD+ -dependent reactions. Certain 3beta-HSD isoforms have been reported to exhibit additional dehydrogenase character (e.g., 17-hydroxysteroid dehydrogenase/reductase). We have investigated whether mouse type I (adrenal/gonadal) and type ...
Bae Gyu-Un - - 2004
We previously reported that hydrogen peroxide (H2O2) mediates mitogen activation of ribosomal protein S6 kinase 1 (S6K1) which plays an important role in cell proliferation and growth. In this study, we investigated a possible role of H2O2 as a molecular linker in Rac1 activation of S6K1. Overexpression of recombinant catalase ...
Chu Chun - - 2004
PURPOSE: Establish and characterize a MDCK cell line that stably expressed PepT1. METHODS: MDCK cells stably transfected with N-terminal HA-tagged hPepT1 were examined by immunostaining using antibodies to HA and hPepT1. HA-hPepT1 expression was verified by immunoprecipitation and Western blotting from cell lysates. The function of the expressed HA-hPepT1 was ...
Hazarika Parul - - 2004
Flotillin 2 (flot-2) is a highly conserved protein isolated from caveolae/lipid raft domains that tether growth factor receptors linked to signal transduction pathways. Flot-2 protein and mRNA were increased in tumorigenic and metastatic melanoma cell lines in vitro, and the immunostaining intensity increased substantially across a tissue array of melanocytic ...
Haas Michael J MJ Division of Endocrinology, Diabetes, and Metabolism, Department of Internal Medicine, Saint Louis University School of Medicine, Saint Louis, MO 63104, - - 2005
The thyroid hormone responsive protein (THRP) is a novel gene product that remains responsive to thyroid hormone (TH) in the cerebral cortex of adult rats. To study the effects of THRP on neuronal cell survival, primary neurons cultured from rats at embryonic day 19 were treated with either 10(-7) mol ...
Liu Y - - 2005
We have studied the in vitro transfection of a plasmid DNA with the lacZ gene to HeLa-S3 cells and hemolysis in a red blood cell (RBC) suspension under pulsed ultrasound with duty cycles gamma of 10, 20 and 30% using a digital sonifier at a frequency of 20 kHz and ...
Yamamoto Kozo - - 2004
The cytolethal distending toxin (Cdt) from Actinobacillus actinomycetemcomitans consists of three proteins, CdtA, CdtB, and CdtC, which are responsible for cell cycle arrest and apoptosis. In the present study, local delivery systems of recombinant CdtB and CdtB-expressing plasmid were established using Ca9-22, human gingival squamous cell carcinoma cell line. When ...
Williams M V - - 2004
A small interfering double stranded RNA molecule (siRNA, 21 bp) corresponding to a portion (nucleotides 337 to 357) of domain 3 of the human dUTPase was synthesized and used to determine whether it could down-regulate dUTPase activity in human cells. Transfection of the siRNA into HeLa and HT29 cells resulted ...
Iczkowski Kenneth A - - 2004
Transfection efficiency of the novel reagent metafectene has not been compared with that of lipofectamine in the published English literature. We used these agents to transfect two prostate cancer cell lines, PC3 and G(s) alpha, with a deoxyribonucleic acid (DNA) expression vector that generates double-stranded ribonucleic acid (RNA) for RNA ...
Zhang Yan - - 2004
The recombinant vector (pLLTK), containing murine serum albumin (ALB) gene promoter/enhancer directing the herpes simplex virus thymidine kinase (HSV-tk) gene expression was constructed to study its effect on hepatic cell-specific damage. Firstly,in order to compare the hepatic cell-specific transcriptional activity, three vectors were constructed in which green fluorescent protein (GFP) ...
Kabilova T O - - 2004
Suppression of c-myc protooncogene expression in KB-3-1 cells by siRNA was investigated. The siRNA duplex targeted to the exon 3 of c-myc mRNA was prepared by in vitro transcription with T7 RNA polymerase on short dsDNA-templates. It was found that incubation of KB-3-1 cells in the presence of 75 nM ...
Aucoin Rebecca - - 2004
Heparanase (HPSE-1) is an endo-beta-D-glucuronidase involved in the degradation of cell-surface/extracellular matrix heparan sulfate (HS) in normal and neoplastic tissues. HPSE-1 represents the first example of purification and cloning of a mammalian HS-degradative enzyme. Elevated HPSE-1 levels are known to be associated with metastatic cancers, directly implicating HPSE-1 in metastatic ...
Shimamura Takeshi - - 2004
Dysadherin is a membrane glycoprotein expressed strongly in several human cancers. Overexpression of dysadherin in tumor cells is closely associated with malignant phenotype (e.g., metastasis) and poor prognosis. In our analysis, six pancreatic cancer cell lines showed a positive correlation between dysadherin expression and cell motility. Introduction of small interfering ...
Ning Shuangli - - 2004
Survivin is recognized as a general target in cancer therapy because of its selective overexpression in the majority of tumors. In bladder cancer (BCa), its expression correlates with tumor grade, recurrence risk and survival. In this study, we compared the therapeutic efficiency of two survivin specific small interfering RNA (siRNA) ...
Ogawa Ryohei - - 2004
PURPOSE: We have previously reported that cell membrane modification by lidocaine or heat can enhance ultrasound mediated transfection (USMT) on PC-3 cells in vitro. In the current study we investigated whether such enhancement could be observed using the T24 human bladder cancer cell line in vitro along with PC-3 in ...
Cui Jing - - 2004
OBJECTIVE: To observe the in vitro expression of DNA vaccine (recombinant eukaryotic expression plasmid pcDNA3-TspE1) encoding a Mr 31,000 antigen of Trichinella spiralis in Chinese hamster ovary (CHO) cells and analyze the antigenicity of the products expressed. METHODS: The recombinant plasmid pcDNA3-TspE1 was transfected into CHO cells by using cationic ...
Zaunbrecher, Gretchen Marie
Together with the advancements in animal cloning, the ability to efficiently target specific genes in somatic cells would greatly enhance several areas of research. While it has been possible for quite some time to target specific genes in the germ cells of mice, the advancements in somatic cell gene targeting ...
Raul Ujwala - - 2004
The cytokeratin (CK) pair 8 and 18 is normally expressed in all simple epithelia. This pair is not normally seen in stratified epithelial cells. Squamous cell carcinomas derived from stratified epithelia show anomalous expression of this CK pair. It is not known whether CKs 8 and 18 in any way ...
Dragomir Anca - - 2004
Cationic liposomes are widely used as gene transfer agents in in vitro and in vivo studies of cystic fibrosis. In this study we report comparative results of cationic mediated transfection in several cell lines. We have tested epithelial cell lines expressing the wild-type cystic fibrosis transmembrane protein CFTR (bronchial epithelium-16HBE14o-, ...
Gupta Mona - - 2004
This study provides a method for enhancing the delivery of nucleic acid molecules to cells by encapsulating it inside the hydrogel pullulan nanoparticles. In this study, pullulan nanoparticles encapsulating pBUDLacZ plasmid has been prepared inside the aqueous droplets of w/o microemulsions. Transmission electron microscopy (TEM) image showed that the particles ...
Schneider Frank - - 2004
We developed an assay allowing the detection and quantification of cell death after transient expression of apoptotic genes in B- and T-lymphocytes. For efficient gene transfer, B- and T-cells were electroporated under optimized conditions. To blind out the high background of non-transfected cells and cell death caused by the electroporation ...
Kato Koichi - - 2004
Cell-based microarrays are emerging as a tool for analyzing the functions of genes in cells. However, partly due to the difficulty of cell immobilization, the application of this method has been limited to adherent cells. We previously reported a method that rapidly and strongly attached living nonadherent cells to glass ...
Erfle Holger - - 2004
RNA interference (RNAi) is a recent advance that provides the possibility to reduce the expression of specific target genes in cultured mammalian cells with potential applications on a genome-wide scale. However, to achieve this, robust methodologies that allow automated and efficient delivery of small interfering RNAs (siRNAs) into living cultured ...
Tuo Xiao-ye - - 2004
OBJECTIVE: To establish a cell line for stable expression of human beta-defensin 3 (hBD3). METHODS: Full length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofectamine. Cell clones survived ...
Rocha A - - 2004
Several methods to improve transfection of epithelioma papulosum cyprini (EPC) carp cells have been tested and are reported here. By modifying the cell cycle state of EPC cell monolayers and selecting the best promoter for the plasmid to be transfected, we increased transfection efficiency from 12.8% to 55.1% and decreased ...
Ma Dengbo - - 2004
Screening of stably transfected cells on multi-well plates is most efficient when a maximum number of wells contain a single colony. The multinomial distribution has been used to derive a novel equation that relates the number of wells containing a specified number of cell-lines and the total number of viable ...
Li Ke-qiang - - 2004
BACKGROUND: This study was designed to obtain a recombinant retroviral vector containing the human hepatocellular carcinoma-related gene ANGPTL4 (angiopoietin-like 4) cDNA and to evaluate the anti-tumor effect of recombinant retroviral vector-mediated human ANGPTL4 gene transfection. METHODS: ANGPTL4 cDNA was cloned in vitro from normal human liver cells HL-7702 by using ...
Wang Zei Wen - - 2004
The thermoregulated T7 expression system was previously reported to be an effective way to produce massive amounts of recombinant proteins (Chao, Y. P.; Law, W. S.; Chen, P. T.; Hung, W. B. High production of heterologous proteins in Escherichia coli using the thermo-regulated T7 expression system. Appl. Microbiol. Biotechnol. 2002b, ...
Kii Isao - - 2004
We studied cadherin-11 function in the differentiation of mesenchymal cells. Teratomas harboring the cadherin-11 gene generated bone and cartilage preferentially. Cadherin-11 transfectants of C2C12 cells and cadherin-11 and/or N-cadherin transfectants of L cells showed that cadherin-11 together with N-cadherin-induced expression of ALP and FGF receptor 2. These results suggest that ...
Matsuyama Syujirou - - 2004
Senescence marker protein-30 (SMP30) is highly expressed in cytosol of hepatocytes, and its amount decreases with aging. Human hepatocellular carcinoma cell line was transfected with pcDNA3/SMP30 (SMP30 transfectants), or as a control with pcDNA3 (mock transfectants). When cells were exposed to 20 ng/ml tumor necrosis factor-alpha (TNF-alpha) plus 10 ng/ml ...
Fellig Yakov - - 2004
Current in vitro models for hepatitis B virus (HBV) are based on human hepatoblastoma cell lines transfected with HBV genome. The objective of this work was to develop an in vitro, hepatocellular carcinoma (HCC)-based system supporting HBV full replication and producing mature viral particles. The FLC4 human HCC cell line ...
Wang Ying - - 2004
Cross-talk between ERalpha and STAT5a was demonstrated to mediate through a direct physical association between the two proteins. By GST pull-down assays and functional assays with various constructs of ERalpha and STAT5a, it was shown that the C-termini of these two proteins were mainly responsible for this interaction. Furthermore, the ...
Krämer Michael - - 2004
Polycationic dendrimers are interesting nonviral vectors for in vitro DNA delivery. We describe a simple approach to the synthesis of dendritic polyamines with different molecular weights and adjustable flexibility (degrees of branching; DB). Both parameters influence the transfection efficiency and the cell toxicity of the polymer. Functionalization of hyperbranched polyethylenimine ...
Tzavelas Christos - - 2004
Previous work from this laboratory has shown that immunoporation has the potential for the selective transfection of a range of different animal cells based on their immunological identity. The unique ability of immunoporation to target cells for transfection combined with the high efficiency of transfection and the high viability of ...
Ciccaglione A R - - 2004
The E1 protein of hepatitis C virus (HCV) shows the ability to induce cell lysis by the alteration of membrane permeability when expressed in Escherichia coli cells. This function seems to be an intrinsic property of a C-terminal hydrophobic region of E1 as permeability changes and cell lysis can be ...
Jin Qi-Huang - - 2004
AIM: To study the potential function of acetylcholinesterase (AChE) in apoptosis through overexpression of AChE in Normal Rat Kidney (NRK) cells. METHODS: AChE activity was detected by the method of Karnovsky and Roots. Activated caspase-3 was analyzed by Western blotting and immunofluorescence with antibody special to activated caspase-3 fragment. The ...
Zhu-Ge Jian - - 2004
AIM: To study the enzyme activity of CYP2C18 variant with exon 5 skipped. METHODS: A full length CYP2C18 cDNA X1 and an exon 5 skipped variant CYP2C18 X2 were separately subcloned into mammalian expression vector pREP9 to transfect HepG2 cells. The expression of CYP2C18 mRNA in transgenic cells and human ...
Xu Xin-bao - - 2004
BACKGROUND: Transforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90. METHODS: The ...
Yoo Hyuk Sang - - 2004
A new cationic emulsion system with high density was prepared increasing in vitro transfection efficiencies of adherent cells. Lipiodol with a density of 1.3 (g/ml) was selected to increase the density of the DNA/emulsion complex. Cationic lipid emulsions were formulated with mixtures of lipiodol and squalene as the oil phase ...
Wang Ji-Shi - - 2004
20-hydroxyeicosatetraenoic acid (20-HETE), a CYP4A-derived arachidonic acid metabolite, is a potent vasoconstrictor and a modulator of vascular reactivity. We have shown that CYP4A1 and CYP4A2 are the major CYP4A isoforms expressed in the rat renal microcirculation. In the present study, we constructed two bicistronic vectors, pIRES2-EGFP-4A1 and pIRES2-EGFP-4A2, and examined ...
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