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Joseph A Frank - One of the best experts on this subject based on the ideXlab platform.
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expression of transferrin receptor and ferritin following ferumoxides Protamine Sulfate labeling of cells implications for cellular magnetic resonance imaging
NMR in Biomedicine, 2006Co-Authors: Edyta Pawelczyk, Ali S Arbab, Sunil D Pandit, Elbert Hu, Joseph A FrankAbstract:Ferumoxides–Protamine Sulfate (FE-Pro) complexes are used for intracellular magnetic labeling of cells to non-invasively monitor cell trafficking by in vivo MRI. FE-Pro labeling is non-toxic to cells; however, the effects of FE-Pro labeling on cellular expression of transferrin receptor (TfR-1) and ferritin, proteins involved in iron transport and storage, has not been reported. FE-Pro-labeled human mesenchymal stem cells (MSCs), HeLa cells and primary macrophages were cultured from 1 week to 2 months and evaluated for TfR-1 and ferritin gene expression by RT-PCR and protein levels were determined using Western blots. MTT (proliferation assay) and reactive oxygen species (ROS) analysis were performed. FE-Pro labeling of HeLa and MSCs resulted in a transient decrease in TfR-1 mRNA and protein levels. In contrast, Fe-Pro labeling of primary macrophages resulted in an increase in TfR-1 mRNA but not in TfR-1 protein levels. Ferritin mRNA and protein levels increased transiently in labeled HeLa and macrophages but were sustained in MSCs. No changes in MTT and ROS analysis were noted. In conclusion, FE-Pro labeling elicited physiological changes of iron metabolism or storage, validating the safety of this procedure for cellular tracking by MRI. Published in 2006 by John Wiley & Sons, Ltd.
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expression of transferrin receptor and ferritin following ferumoxides Protamine Sulfate labeling of cells implications for cellular magnetic resonance imaging
NMR in Biomedicine, 2006Co-Authors: Edyta Pawelczyk, Ali S Arbab, Sunil D Pandit, Joseph A FrankAbstract:Ferumoxides-Protamine Sulfate (FE-Pro) complexes are used for intracellular magnetic labeling of cells to non-invasively monitor cell trafficking by in vivo MRI. FE-Pro labeling is non-toxic to cells; however, the effects of FE-Pro labeling on cellular expression of transferrin receptor (TfR-1) and ferritin, proteins involved in iron transport and storage, has not been reported. FE-Pro-labeled human mesenchymal stem cells (MSCs), HeLa cells and primary macrophages were cultured from 1 week to 2 months and evaluated for TfR-1 and ferritin gene expression by RT-PCR and protein levels were determined using Western blots. MTT (proliferation assay) and reactive oxygen species (ROS) analysis were performed. FE-Pro labeling of HeLa and MSCs resulted in a transient decrease in TfR-1 mRNA and protein levels. In contrast, Fe-Pro labeling of primary macrophages resulted in an increase in TfR-1 mRNA but not in TfR-1 protein levels. Ferritin mRNA and protein levels increased transiently in labeled HeLa and macrophages but were sustained in MSCs. No changes in MTT and ROS analysis were noted. In conclusion, FE-Pro labeling elicited physiological changes of iron metabolism or storage, validating the safety of this procedure for cellular tracking by MRI.
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labeling of cells with ferumoxides Protamine Sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells
NMR in Biomedicine, 2005Co-Authors: Ali S Arbab, Gene T Yocum, Aarif Y Khakoo, Elizabeth J Read, Ali M Rad, Vicki Fellowes, Joseph A FrankAbstract:Two FDA-approved agents, ferumoxides (Feridex), a suspension of superparamagnetic iron oxide (SPIO) nanoparticles and Protamine Sulfate, a drug used to reverse heparin anticoagulation, can be complexed and used to label cells magnetically ex vivo. Labeling stem cells with ferumoxides-Protamine Sulfate (FePro) complexes allows for non-invasive monitoring by MRI. However, in order for stem cell trials or therapies to be effective, this labeling technique must not inhibit the ability of cells to differentiate. In this study, we examined the effect of FePro labeling on stem cell differentiation. Viability, phenotypic expression and differential capacity of FePro labeled CD34 + hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) were compared with unlabeled control cells. Colony-forming unit (CFU) assays showed that the capacity to differentiate was equivalent for labeled and unlabeled HSC. Furthermore, labeling did not alter expression of surface phenotypic markers (CD34, CD31, CXCR4, CD20, CD3 and CD14) on HSC, as measured by flow cytometry. SDF-1-induced HSC migration and HSC differentiation to dendritic cells were also unaffected by FePro labeling. Both FePro-labeled and unlabeled MSC were cultured in chondrogenesis-inducing conditions. Alcian blue staining for proteoglycans revealed similar chondrogenic differentiation for both FePro-labeled and unlabeled cells. Furthermore, collagen X proteins, indicators of cartilage formation, were detected at similar levels in both labeled and unlabeled cell pellets. Prussian blue staining confirmed that cells in labeled pellets contained iron oxide, whereas cells in unlabeled pellets did not. It is concluded that FePro labeling does not alter the function or differentiation capacity of HSC and MSC. These data increase confidence that MRI studies of FePro-labeled HSC or MSC will provide an accurate representation of in vivo trafficking of unlabeled cells.
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efficient magnetic cell labeling with Protamine Sulfate complexed to ferumoxides for cellular mri
Blood, 2004Co-Authors: Ali S Arbab, Gene T Yocum, Heather Kalish, Elaine K Jordan, Stasia A Anderson, Aarif Y Khakoo, Elizabeth J Read, Joseph A FrankAbstract:Recently, there have been several reports using various superparamagnetic iron oxide (SPIO) nanoparticles to label mammalian cells for monitoring their temporal and spatial migration in vivo by magnetic resonance imaging (MRI). The purpose of this study was to evaluate the efficiency and toxicity of labeling cells using 2 commercially available Food and Drug Administration (FDA)-approved agents, ferumoxides, a suspension of dextran-coated SPIO used as an MRI contrast agent, and Protamine Sulfate, conventionally used to reverse heparin anticoagulation but also used ex vivo as a cationic transfection agent. After labeling of human mesenchymal stem cells (MSCs) and hematopoietic (CD34+) stem cells and other mammalian cells with ferumoxides-Protamine Sulfate complexes (FE-Pro), cellular toxicity, functional capacity, and quantitative cellular iron incorporation were determined. FE-Pro-labeled cells demonstrated no short- or long-term toxicity, changes in differentiation capacity of the stem cells, or changes in phenotype when compared with unlabeled cells. Efficient labeling with FE-Pro was observed with iron content per cell varying between 2.01 ± 0.1 pg for CD34+ cells and 10.94 ± 1.86 pg for MSCs with 100% of cells labeled. Cell labeling using these agents should facilitate the translation of this method to clinical trials for evaluation of trafficking of infused or transplanted cells by MRI. (Blood. 2004;104:1217-1223)
Ali S Arbab - One of the best experts on this subject based on the ideXlab platform.
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effects of ferumoxides Protamine Sulfate labeling on immunomodulatory characteristics of macrophage like thp 1 cells
PLOS ONE, 2008Co-Authors: Branislava Janic, Ali M Rad, A S M Iskander, Hamid Soltanianzadeh, Ali S ArbabAbstract:Superparamagnetic Iron Oxide (SPIO) complexed with cationic transfection agent is used to label various mammalian cells. Labeled cells can then be utilized as an in vivo magnetic resonance imaging (MRI) probes. However, certain number of in vivo administered labeled cells may be cleared from tissues by the host's macrophages. For successful translation to routine clinical application of SPIO labeling method it is important that this mode of in vivo clearance of iron does not elicit any diverse immunological effects. The purpose of this study was to demonstrate that SPIO agent ferumoxides-Protamine Sulfate (FePro) incorporation into macrophages does not alter immunological properties of these cells with regard to differentiation, chemotaxis, and ability to respond to the activation stimuli and to modulate T cell response. We used THP-1 cell line as a model for studying macrophage cell type. THP-1 cells were magnetically labeled with FePro, differentiated with 100 nM of phorbol ester, 12-Myristate-13-acetate (TPA) and stimulated with 100 ng/ml of LPS. The results showed 1) FePro labeling had no effect on the changes in morphology and expression of cell surface proteins associated with TPA induced differentiation; 2) FePro labeled cells responded to LPS with slightly higher levels of NFκB pathway activation, as shown by immunobloting; TNF-α secretion and cell surface expression levels of CD54 and CD83 activation markers, under these conditions, were still comparable to the levels observed in non-labeled cells; 3) FePro labeling exhibited differential, chemokine dependent, effect on THP-1 chemotaxis with a decrease in cell directional migration to MCP-1; 4) FePro labeling did not affect the ability of THP-1 cells to down-regulate T cell expression of CD4 and CD8 and to induce T cell proliferation. Our study demonstrated that intracellular incorporation of FePro complexes does not alter overall immunological properties of THP-1 cells. The described experiments provide the model for studying the effects of in vivo clearance of iron particles via incorporation into the host's macrophages that may follow after in vivo application of any type of magnetically labeled mammalian cells. To better mimic the complex in vivo scenario, this model may be further exploited by introducing additional cellular and biological, immunologically relevant, components.
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expression of transferrin receptor and ferritin following ferumoxides Protamine Sulfate labeling of cells implications for cellular magnetic resonance imaging
NMR in Biomedicine, 2006Co-Authors: Edyta Pawelczyk, Ali S Arbab, Sunil D Pandit, Elbert Hu, Joseph A FrankAbstract:Ferumoxides–Protamine Sulfate (FE-Pro) complexes are used for intracellular magnetic labeling of cells to non-invasively monitor cell trafficking by in vivo MRI. FE-Pro labeling is non-toxic to cells; however, the effects of FE-Pro labeling on cellular expression of transferrin receptor (TfR-1) and ferritin, proteins involved in iron transport and storage, has not been reported. FE-Pro-labeled human mesenchymal stem cells (MSCs), HeLa cells and primary macrophages were cultured from 1 week to 2 months and evaluated for TfR-1 and ferritin gene expression by RT-PCR and protein levels were determined using Western blots. MTT (proliferation assay) and reactive oxygen species (ROS) analysis were performed. FE-Pro labeling of HeLa and MSCs resulted in a transient decrease in TfR-1 mRNA and protein levels. In contrast, Fe-Pro labeling of primary macrophages resulted in an increase in TfR-1 mRNA but not in TfR-1 protein levels. Ferritin mRNA and protein levels increased transiently in labeled HeLa and macrophages but were sustained in MSCs. No changes in MTT and ROS analysis were noted. In conclusion, FE-Pro labeling elicited physiological changes of iron metabolism or storage, validating the safety of this procedure for cellular tracking by MRI. Published in 2006 by John Wiley & Sons, Ltd.
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expression of transferrin receptor and ferritin following ferumoxides Protamine Sulfate labeling of cells implications for cellular magnetic resonance imaging
NMR in Biomedicine, 2006Co-Authors: Edyta Pawelczyk, Ali S Arbab, Sunil D Pandit, Joseph A FrankAbstract:Ferumoxides-Protamine Sulfate (FE-Pro) complexes are used for intracellular magnetic labeling of cells to non-invasively monitor cell trafficking by in vivo MRI. FE-Pro labeling is non-toxic to cells; however, the effects of FE-Pro labeling on cellular expression of transferrin receptor (TfR-1) and ferritin, proteins involved in iron transport and storage, has not been reported. FE-Pro-labeled human mesenchymal stem cells (MSCs), HeLa cells and primary macrophages were cultured from 1 week to 2 months and evaluated for TfR-1 and ferritin gene expression by RT-PCR and protein levels were determined using Western blots. MTT (proliferation assay) and reactive oxygen species (ROS) analysis were performed. FE-Pro labeling of HeLa and MSCs resulted in a transient decrease in TfR-1 mRNA and protein levels. In contrast, Fe-Pro labeling of primary macrophages resulted in an increase in TfR-1 mRNA but not in TfR-1 protein levels. Ferritin mRNA and protein levels increased transiently in labeled HeLa and macrophages but were sustained in MSCs. No changes in MTT and ROS analysis were noted. In conclusion, FE-Pro labeling elicited physiological changes of iron metabolism or storage, validating the safety of this procedure for cellular tracking by MRI.
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labeling of cells with ferumoxides Protamine Sulfate complexes does not inhibit function or differentiation capacity of hematopoietic or mesenchymal stem cells
NMR in Biomedicine, 2005Co-Authors: Ali S Arbab, Gene T Yocum, Aarif Y Khakoo, Elizabeth J Read, Ali M Rad, Vicki Fellowes, Joseph A FrankAbstract:Two FDA-approved agents, ferumoxides (Feridex), a suspension of superparamagnetic iron oxide (SPIO) nanoparticles and Protamine Sulfate, a drug used to reverse heparin anticoagulation, can be complexed and used to label cells magnetically ex vivo. Labeling stem cells with ferumoxides-Protamine Sulfate (FePro) complexes allows for non-invasive monitoring by MRI. However, in order for stem cell trials or therapies to be effective, this labeling technique must not inhibit the ability of cells to differentiate. In this study, we examined the effect of FePro labeling on stem cell differentiation. Viability, phenotypic expression and differential capacity of FePro labeled CD34 + hematopoietic stem cells (HSC) and mesenchymal stem cells (MSC) were compared with unlabeled control cells. Colony-forming unit (CFU) assays showed that the capacity to differentiate was equivalent for labeled and unlabeled HSC. Furthermore, labeling did not alter expression of surface phenotypic markers (CD34, CD31, CXCR4, CD20, CD3 and CD14) on HSC, as measured by flow cytometry. SDF-1-induced HSC migration and HSC differentiation to dendritic cells were also unaffected by FePro labeling. Both FePro-labeled and unlabeled MSC were cultured in chondrogenesis-inducing conditions. Alcian blue staining for proteoglycans revealed similar chondrogenic differentiation for both FePro-labeled and unlabeled cells. Furthermore, collagen X proteins, indicators of cartilage formation, were detected at similar levels in both labeled and unlabeled cell pellets. Prussian blue staining confirmed that cells in labeled pellets contained iron oxide, whereas cells in unlabeled pellets did not. It is concluded that FePro labeling does not alter the function or differentiation capacity of HSC and MSC. These data increase confidence that MRI studies of FePro-labeled HSC or MSC will provide an accurate representation of in vivo trafficking of unlabeled cells.
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efficient magnetic cell labeling with Protamine Sulfate complexed to ferumoxides for cellular mri
Blood, 2004Co-Authors: Ali S Arbab, Gene T Yocum, Heather Kalish, Elaine K Jordan, Stasia A Anderson, Aarif Y Khakoo, Elizabeth J Read, Joseph A FrankAbstract:Recently, there have been several reports using various superparamagnetic iron oxide (SPIO) nanoparticles to label mammalian cells for monitoring their temporal and spatial migration in vivo by magnetic resonance imaging (MRI). The purpose of this study was to evaluate the efficiency and toxicity of labeling cells using 2 commercially available Food and Drug Administration (FDA)-approved agents, ferumoxides, a suspension of dextran-coated SPIO used as an MRI contrast agent, and Protamine Sulfate, conventionally used to reverse heparin anticoagulation but also used ex vivo as a cationic transfection agent. After labeling of human mesenchymal stem cells (MSCs) and hematopoietic (CD34+) stem cells and other mammalian cells with ferumoxides-Protamine Sulfate complexes (FE-Pro), cellular toxicity, functional capacity, and quantitative cellular iron incorporation were determined. FE-Pro-labeled cells demonstrated no short- or long-term toxicity, changes in differentiation capacity of the stem cells, or changes in phenotype when compared with unlabeled cells. Efficient labeling with FE-Pro was observed with iron content per cell varying between 2.01 ± 0.1 pg for CD34+ cells and 10.94 ± 1.86 pg for MSCs with 100% of cells labeled. Cell labeling using these agents should facilitate the translation of this method to clinical trials for evaluation of trafficking of infused or transplanted cells by MRI. (Blood. 2004;104:1217-1223)
Goksel şener - One of the best experts on this subject based on the ideXlab platform.
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protective effects of taurine on Protamine Sulfate induced bladder damage
World Journal of Urology, 2006Co-Authors: Ali Zeybek, şule Cetinel, Feriha Ercan, Beyhan Saglam, Esra Cikler, Goksel şenerAbstract:The present study was designed to investigate the putative protective effects of taurine on Protamine Sulfate (PS) induced bladder injury. Wistar albino female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or PS (PS group) dissolved in phosphate buffered solution. In the PS + taurine (PS+Tau) group, after the PS instillation, taurine (50 mg/kg) was injected intraperitoneally for 3 days. Histopathological changes were investigated by light and scanning electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde (MDA) (a biomarker of oxidative damage) and glutathione (GSH) (a biomarker of protective oxidative injury) levels. In the PS group ulcerated areas, an irregular mucus layer, inflammatory cell infiltration, and increased number of mast cells were observed. In the PS+Tau group, a relatively normal urothelial topography, glycosaminoglycan layer, and decreased number of mucosal mast cells and inflammatory cells were observed. Increased MDA levels as a result of PS induction lead us to propose that free radicals may have a critical role in this injury. The significant decrease in MDA and increase in GSH levels in the PS+Tau group compared to PS group was in accordance with morphological findings. Based on the results, taurine treatment significantly prevented PS induced degenerative morphological and biochemical changes of urinary bladder mucosa.
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the ameliorating effect of melatonin on Protamine Sulfate induced bladder injury and its relationship to interstitial cystitis
The Journal of Urology, 2003Co-Authors: şule Cetinel, Feriha Ercan, Serap şirvanci, Ozer şehirli, Yasemin Ersoy, Tangul şan, Goksel şenerAbstract:ABSTRACTPurpose: The pineal hormone melatonin was recently shown to have free radical scavenging ability and it reduces lipid peroxidation. In this morphological study we investigated the effects of melatonin on Protamine Sulfate (Sigma Chemical Co., St. Louis, Missouri) induced bladder injury.Materials and Methods: Albino Wistar female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or Protamine Sulfate (bladder injury group) dissolved in phosphate buffered solution. In the Protamine Sulfate plus melatonin group after Protamine Sulfate instillation melatonin was injected intraperitoneally. Bladder morphology was investigated by light and electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde levels.Results: In the bladder injury group ulcerated areas, an irregular glycosaminoglycan layer, increased number of mast cells, vacuole formation, dilated perinuclear cistern, formation of pleomorphic and uniform microvilli, an...
şule Cetinel - One of the best experts on this subject based on the ideXlab platform.
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protective effects of taurine on Protamine Sulfate induced bladder damage
World Journal of Urology, 2006Co-Authors: Ali Zeybek, şule Cetinel, Feriha Ercan, Beyhan Saglam, Esra Cikler, Goksel şenerAbstract:The present study was designed to investigate the putative protective effects of taurine on Protamine Sulfate (PS) induced bladder injury. Wistar albino female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or PS (PS group) dissolved in phosphate buffered solution. In the PS + taurine (PS+Tau) group, after the PS instillation, taurine (50 mg/kg) was injected intraperitoneally for 3 days. Histopathological changes were investigated by light and scanning electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde (MDA) (a biomarker of oxidative damage) and glutathione (GSH) (a biomarker of protective oxidative injury) levels. In the PS group ulcerated areas, an irregular mucus layer, inflammatory cell infiltration, and increased number of mast cells were observed. In the PS+Tau group, a relatively normal urothelial topography, glycosaminoglycan layer, and decreased number of mucosal mast cells and inflammatory cells were observed. Increased MDA levels as a result of PS induction lead us to propose that free radicals may have a critical role in this injury. The significant decrease in MDA and increase in GSH levels in the PS+Tau group compared to PS group was in accordance with morphological findings. Based on the results, taurine treatment significantly prevented PS induced degenerative morphological and biochemical changes of urinary bladder mucosa.
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the ameliorating effect of melatonin on Protamine Sulfate induced bladder injury and its relationship to interstitial cystitis
The Journal of Urology, 2003Co-Authors: şule Cetinel, Feriha Ercan, Serap şirvanci, Ozer şehirli, Yasemin Ersoy, Tangul şan, Goksel şenerAbstract:ABSTRACTPurpose: The pineal hormone melatonin was recently shown to have free radical scavenging ability and it reduces lipid peroxidation. In this morphological study we investigated the effects of melatonin on Protamine Sulfate (Sigma Chemical Co., St. Louis, Missouri) induced bladder injury.Materials and Methods: Albino Wistar female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or Protamine Sulfate (bladder injury group) dissolved in phosphate buffered solution. In the Protamine Sulfate plus melatonin group after Protamine Sulfate instillation melatonin was injected intraperitoneally. Bladder morphology was investigated by light and electron microscopy. Tissue samples were also obtained to determine bladder malondialdehyde levels.Results: In the bladder injury group ulcerated areas, an irregular glycosaminoglycan layer, increased number of mast cells, vacuole formation, dilated perinuclear cistern, formation of pleomorphic and uniform microvilli, an...
Leaf Huang - One of the best experts on this subject based on the ideXlab platform.
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intravesical Protamine Sulfate and potassium chloride as a model for bladder hyperactivity
Urology, 2003Co-Authors: Yao Chi Chuang, Leaf Huang, Michael B Chancellor, Satoshi Seki, Naoki Yoshimura, Pradeep Tyagi, John P Lavelle, William C De Groat, Matthew O FraserAbstract:Abstract Objectives An acute animal model for hyperactive bladder in rats was developed using intravesical infusion of Protamine Sulfate (PS), an agent thought to break down urothelial barrier function, and physiologic concentrations of potassium chloride (KCl). Methods Continuous cystometrograms (CMGs) were performed in urethane-anesthetized female rats by filling the bladder (0.04 mL/min) with normal saline followed by intravesical infusion of a test solution consisting of either KCl (100 or 500 mM) or PS (10 or 30 mg/mL) for 60 minutes. Subsequently, the 10 mg/mL PS-treated animals were infused intravesically with 100, 300, or 500 mM KCl. Some animals were pretreated with capsaicin (125 mg/kg subcutaneously) 4 days before the experiments. Results Unlike KCl (100 or 500 mM) or a low concentration of PS (10 mg/mL) alone, the intravesical administration of a high concentration of PS (30 mg/mL) produced irritative effects with a decreased intercontraction interval (by 80.6%). After infusion of a low concentration of PS, infusion of 300 or 500 mM KCl produced irritative effects (intercontraction interval decreased by 76.9% or 82.9%, respectively). The onset of irritation occurred more rapidly after 500 mM KCl (10 to 15 minutes) than after 300 mM KCl (20 to 30 minutes). Capsaicin pretreatment delayed the onset (approximately 60 minutes) and reduced the magnitude (intercontraction interval decreased by 35.5%) of irritative effects. Conclusions Intravesical administration of KCl after PS treatment activates capsaicin-sensitive afferents and detrusor muscle and presumably capsaicin-resistant afferents. Modest, noncytotoxic affronts to urothelial barrier function can result in dramatic irritative responses. This model may be useful in the study of bladder irritation and hyperactivity.
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Protamine Sulfate enhances lipid mediated gene transfer
Gene Therapy, 1997Co-Authors: F Sorgi, Sanchita Bhattacharya, Leaf HuangAbstract:A polycationic peptide, Protamine Sulfate, USP, has been shown to be able to condense plasmid DNA efficiently for delivery into several different types of cells in vitro by several different types of cationic liposomes. The monovalent cationic liposomal formulations (DC-Chol and lipofectin) exhibited increased transfection activities comparable to that seen with the multivalent cationic liposome formulation, lipofectamine. This suggests that lipofectamine’s superior in vitro activity arises from its ability to condense DNA efficiently and that Protamine’s primary role is that of a condensation agent, although it also possesses several amino acid sequences resembling that of a nuclear localization signal. While the use of polycations to condense DNA has been previously reported, the use of Protamine Sulfate, USP as a condensation agent was found to be superior to poly- L-lysine as well as to various other types of Protamine. These differences among various salt forms of Protamine appear to be attributable to structural differences between the Protamines and not due to differences in the net charge of the molecule. The appearance of lysine residues within the Protamine molecule correlate with a reduction in binding affinity to plasmid DNA, as well as an observed loss in transfection-enhancing activity. This finding sheds light on the structural requirements of condensation agents for use in gene transfer protocols. Furthermore, Protamine Sulfate, USP is an FDA-approved compound with a documented safety profile and could be readily used as an adjuvant to a human gene therapy protocol.
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Protamine Sulfate provides enhanced and reproducible intravenous gene transfer by cationic liposome dna complex
Journal of Liposome Research, 1997Co-Authors: Leaf HuangAbstract:AbstractA novel lipid/polycation/DNA (LPD) formulation has been developed for in vivo gene transfer. It involves the condensation of plasmid DNA with Protamine Sulfate, a cationic polypeptide, followed by the addition of DOTAP cationic liposomes. Compared with DOTAP/DNA complex, LPD offers greater protection of plasmid DNA against enzymatic digestion and gives consistently higher gene expression in mice via tail vein injection. The in vivo efficiency of LPD was dependent upon charge ratio and was also affected by the lipid used. Increasing the amount of DNA delivered induced an increase in gene expression. The optimal dose was approximately 50 μg per mouse, at which concentration approximately 10 ng luciferase protein per mg extracted tissue protein could be detected in the lung. Gene expression in the lung was detected as early as 1 h after injection, peaked at 6 h, and declined thereafter. Using LacZ as a reporter gene, it was shown that endothelial cells were the primary locus of transgene expression i...
