Biomedical subjects

Sung Won Choi

Publications and source records attributed to Sung Won Choi.

10 recordsLinked to original sources

PubMed2007

Spectroscopic investigation of cationic comb-type copolymers/DNA interaction: interpolyelectrolyte complex enhancement synchronized with DNA hybridization.

We have demonstrated that cationic comb-type copolymers consisting of a polycation backbone and abundant grafts of water-soluble polymers stabilize DNA hybrids. Furthermore, the copolymers were found to accelerate strand exchange reaction between a double-stranded DNA and its complementary single-stranded DNA. In this article, we investigated the effects of PLL-g-Dex on base pairs of a self-complementary DNA octamer, d(GGAATTCC). The soluble interpolyelectrolyte complex (IPEC) between the DNA and copolymer allowed us to characterize the complex by using spectroscopic methods under physiological ionic condition. Chemical shifts of nucleobase proton signals were not changed by PLL-g-Dex. Furthermore, the copolymer slightly changed the von't Hoff DeltaH accompanying the helix-coil transition of the octamer. These results indicated that the base pairs of the duplex DNA in the IPEC were not perturbed by the polycationic copolymer. It was obviously shown by temperature dependencies of proton and phosphorus NMR spectra that DNA/copolymer interaction was considerably enhanced in response to ds DNA formation. An increase in the density and total number of DNA negative charges upon hybrid formation likely caused the higher affinity of the copolymer with the ds form over that of the copolymer with the ss form. The IPEC formation of CCCs with DNA, however, seems highly sensitive to the coil-helix transition of the DNA.

Cations↗

PubMed2007

Cationic comb-type copolymers for boosting DNA-fueled nanomachines.

For the better applications and developments of DNA nanomachines, their responding kinetics, output, and sequence-selectivity need to be improved. Furthermore, the DNA nanomachines currently have several limitations in operating conditions. Here we show that a simple addition of a cationic comb-type copolymer, poly(l-lysine)-graft-dextran, produces the robust and quick responses of DNA nanomachines under moderate conditions including physiologically relevant conditions even at very low strand concentrations (nanomoles per liter range) through hybrid stabilization and DNA strand exchange acceleration.

Base Sequence↗

PubMed2006

Structural effect of cationic copolymers on nucleic acid-chaperoning activity.

In order to evaluate the effect of cationic copolymer structures on their nucleic acid-chaperoning activity, we prepared various copolymers having different cationic residues or backbone molecular weight. It was revealed that nucleic acid-chaperoning activity increases with increasing molecular weight of the copolymer backbone and that the copolymer having the guanidino groups is effective for increasing nucleic acid-chaperoning activity. Compared with PLL-g-Dex, GPLL-g-Dex has weak activity to stabilize ds DNA. This weak stabilization effect of GPLL-g-Dex may contribute to the higher accelerating effect.

DNA↗

PubMed2005

Mutant tamm-horsfall glycoprotein accumulation in endoplasmic reticulum induces apoptosis reversed by colchicine and sodium 4-phenylbutyrate.

As a consequence of uromodulin gene mutations, individuals develop precocious hyperuricemia, gout, and progressive renal failure. In vitro studies suggest that pathologic accumulation of uromodulin/Tamm-Horsfall glycoprotein (THP) occurs in the endoplasmic reticulum (ER), but the pathophysiology of renal damage is unclear. It was hypothesized that programmed cell death triggered by accumulation of misfolded THP in the ER causes progressive renal disease. Stably transfected human embryonic kidney 293 cells and immortalized thick ascending limb of Henle's loop cells with wild-type and mutated uromodulin cDNA were evaluated to test this hypothesis. Immunocytochemistry, ELISA, and deglycosylation studies indicated that accumulation of mutant THP occurred in the ER. FACS analyses showed a significant increase in early apoptosis signal in human embryonic kidney 293 and thick ascending limb of Henle's loop cells that were transfected with mutant uromodulin constructs. Colchicine and sodium 4-phenylbutyrate treatment increased secretion of THP from the ER to the cell membrane and into the culture media and significantly improved cell viability. These findings indicate that intracellular accumulation of THP facilitates apoptosis and that this may provide the pathologic mechanism responsible for the progressive renal damage associated with uromodulin gene mutations. Colchicine and sodium 4-phenylbutyrate reverse these processes and could potentially be beneficial in ameliorating the progressive renal damage in uromodulin-associated kidney diseases.

Apoptosis↗

PubMed2005

Proteolysis of macrophage inflammatory protein-1alpha isoforms LD78beta and LD78alpha by neutrophil-derived serine proteases.

Macrophage inflammatory protein-1alpha (MIP-1alpha) is a chemokine that leads to leukocyte recruitment and activation at sites of infection. Controlling chemokine activity at sites of infection is important, since excess accumulation of leukocytes may contribute to localized tissue damage. Neutrophil-derived serine proteases modulate the bioactivity of chemokine and cytokine networks through proteolytic cleavage. Because MIP-1alpha is temporally expressed with neutrophils at sites of infection, we examined proteolysis of MIP-1alpha in vitro by the neutrophil-derived serine proteases: cathepsin G, elastase, and proteinase 3. Recombinant human MIP-1alpha isoforms LD78beta and LD78alpha were expressed and purified, and the protease cleavage sites were analyzed by mass spectrometry and peptide sequencing. Chemotactic activities of parent and cleavage molecules were also compared. Both LD78beta and LD78alpha were cleaved by neutrophil lysates at Thr16-Ser17, Phe24-Ile25, Tyr28-Phe29, and Thr31-Ser32. This degradation was inhibited by serine protease inhibitors phenylmethylsulfonyl fluoride and 4-(2-aminoethyl)-benzenesulfonyl fluoride. Incubation of the substrates with individual proteases revealed that cathepsin G preferentially cleaved at Phe24-Ile25 and Tyr28-Phe29, whereas elastase and proteinase 3 cleaved at Thr16-Ser17 and Thr31-Ser32. Proteolysis of LD78beta resulted in loss of chemotactic activity. The role of these proteases in LD78beta and LD78alpha degradation was confirmed by incubation with neutrophil lysates from Papillon-Lefevre syndrome patients, demonstrating that the cell lysates containing inactivated serine proteases could not degrade LD78beta and LD78alpha. These findings suggest that severe periodontal tissue destruction in Papillon-Lefevre syndrome may be related to excess accumulation of LD78beta and LD78alpha and dysregulation of the microbial-induced inflammatory response in the periodontium.

Amino Acid Sequence↗

PubMed2005

The molecular structure effect of cationic comb-type copolymers on nucleic acid chaperone activity.

We have shown that the cationic comb-type copolymers (CCCs) accelerate DNA hybridization and increase stability of DNA duplexes and triplexes. The CCCs were considered to act as "nucleic acid chaperones," promoting the formation of the most stable hybrids. In this study, CCCs with primary amino or guanidino groups were prepared to evaluate the effects of different cationic moieties upon the nucleic acid chaperone activity. CCCs having guanidino groups have higher accelerating effect on DNA strand exchange reactions than that having primary amino groups. It was suggested that some modes of interactions, such as hydrogen bonding, other than ionic interactions between the copolymers and DNAs may be involved upon the strand exchange activation.

DNA↗

PubMed2004

Evaluation of comb-type copolymers with different cationic groups as artificial nucleic acid chaperones.

We have demonstrated that cationic comb-type copolymers (CCCs) accelerate DNA hybridization and increase stability of DNA duplexes and triplexes. The CCCs were considered to act as nucleic acid chaperones that catalyze structural transitions of nucleic acid hybrids. In this study, CCCs with primary amino or guanidino groups were prepared to evaluate the effects of different cationic moieties upon the nucleic acid chaperoning activity. CCCs having guanidino groups have higher accelerating effect on DNA strand exchange reactions (SFRs) than that having primary amino groups. It was suggested that some modes of interactions, such as hydrogen bonding, other than ionic interactions between the copolymers and DNAs may be involved upon the strand exchange activation.

DNA↗

PubMed2004

Preparation of cationic comb-type copolymer having guanidino moieties and its interaction with DNAs.

In order to evaluatet effects of different cationic moieties, such as primary amino and guanidino groups, on interactions between DNAs and cationic comb-type copolymers, comb-type copolymers having guanidino groups were prepared. The copolymers (GPLL-g-Dex) were obtained by guanidination of poly(L-lysine)-graft-dextran copolymers (PLL-g-Dex) using 1-guanyl-3,5-dimethylpyrazole nitrate under weak basic conditions. The resulting copolymers were characterized using NMR spectroscopy and size-exclusion chromatography-multiangle light scattering (SEC-MALS). The primary amino groups of the PLL backbones were thoroughly replaced with guanidino ones without any detectable side reactions, including fragmentation of PLL or Dex chains. The interactions of GPLL-g-Dex and PLL-g-Dex with DNAs were assessed by UV-melting curve measurements. These copolymers diversly affected the melting behavior of double-stranded DNA (dsDNA). GPLL-g-Dex has a lower ability to increase the Tm of dsDNA than PLL-g-Dex and also exhibits a higher affinity for dsDNA. The results suggest that the stability of dsDNA may be affected not only by ionic interaction between the copolymers and DNAs, but also by other modes of interactions, such as hydrogen-bondinginteractions.

Base Sequence↗

PubMed2003

Elastic biodegradable poly(glycolide-co-caprolactone) scaffold for tissue engineering.

Cyclic mechanical strain has been demonstrated to enhance the development and function of engineered smooth muscle (SM) tissues, and it would be necessary for the development of the elastic scaffolds if one wishes to engineer SM tissues under cyclic mechanical loading. This study reports on the development of an elastic scaffold fabricated from a biodegradable polymer. Biodegradable poly(glycolide-co-caprolactone) (PGCL) copolymer was synthesized from glycolide and epsilon-caprolactone in the presence of stannous octoate as catalyst. The copolymer was characterized by (1)H-NMR, gel permeation chromatography and differential scanning calorimetry. Scaffolds for tissue engineering applications were fabricated from PGCL copolymer using the solvent-casting and particle-leaching technique. The PGCL scaffolds produced in this fashion had open pore structures (average pore size = 250 microm) without the usual nonporous skin layer on external surfaces. Mechanical testing revealed that PGCL scaffolds were far more elastic than poly(lactic-co-glycolic acid) (PLGA) scaffolds fabricated using the same method. Tensile mechanical tests indicated that PGCL scaffolds could withstand an extension of 250% without cracking, which was much higher than withstood by PLGA scaffolds (10-15%). In addition, PGCL scaffolds achieved recoveries exceeding 96% at applied extensions of up to 230%, whereas PLGA scaffolds failed (cracked) at an applied strain of 20%. Dynamic mechanical tests showed that the permanent deformation of the PGCL scaffolds in a dry condition produced was less than 4% of the applied strain, when an elongation of 20% at a frequency of 1 Hz (1 cycle per second) was applied for 6 days. Moreover, PGCL scaffolds in a buffer solution also had permanent deformations less than 5% of the applied strain when an elongation of 10% at a frequency of 1 Hz was applied for 2 days. The usefulness of the PGCL scaffolds was demonstrated by engineering SM tissues in vivo. This study shows that the elastic PGCL scaffolds produced in this study could be used to engineer SM-containing tissues (e.g. blood vessels and bladders) in mechanically dynamic environments.

Absorbable Implants↗

PubMed2002

Investigation of DNA strand exchange mechanisms mediated by cationic polymers.

It was observed that conjugation of dextran for PLL provided PLL-g-Dex with new properties of enhancing and accelerating inter-nucleotide recognition (1, 2, 3). In this paper, we report the effects of cationic PLL-g-Dex copolymers on base-pair formation of self-complementary octamer oligonucleotide by using NMR and CD analysis. NMR studies revealed that PLL-g-Dex copolymer did not affect hydrogen-bondings between bases in DNA duplex. CD studies also supported inert nature of the copolymer upon hybrid structure.

Base Sequence↗