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Biomedical subjects

Hyeon Yong Lee

Publications and source records attributed to Hyeon Yong Lee.

At least 19 recordsLinked to original sources

Preparation and characterization of chitosan/gelatin microcapsules containing triclosan.

Chitosan/gelatin (C/G) microcapsules containing triclosan were prepared by a spray drying method. The core material, triclosan (TS) dissolved in octyl salicylate (OS), were emulsified in an aqueous solution containing variable ratios of chitosan/gelatin. The microcapsules were obtained by spray-drying the emulsions. On the scanning electron micrographs, the microcapsules were spherical and exhibited a core and shell morphology. The thermograms of the microcapsules showed no evidence for the melting of TS, suggesting that TS remained dissolved in the cores of the microcapsules and did not exist as a solid crystalline even after dry microcapsules were formed. According to the results of microelectrophoresis study, the point of zero charge of the microcapsules occurred around pH 9.0 and a higher content of chitosan in the microcapsule wall resulted in a higher positive charge of zeta potential. The degree of release of TS and OS from the C/G microcapsules in an aqueous solution of hydroxypropyl-beta-cyclodextrin (HP-beta-CD) was investigated. When chitosan is included in the wall of microcapsules, the degree of release was suppressed. This indicates that chitosan forms a more compact wall than gelatin. On the other hand, TS was released much more than OS. The preferred release of TS is probably due to the higher solubility of TS in the HP-beta-CD solution.

Calorimetry, Differential Scanning↗

In vivo gene therapy of type I diabetic mellitus using a cationic emulsion containing an Epstein Barr Virus (EBV) based plasmid vector.

A cationic emulsion containing an insulin expression plasmid was prepared for the treatment of type 1 diabetic mellitus (DM) in vivo. A rat proinsulin-1 gene was inserted to EBV-based plasmid vectors containing CAG promoter. Cationic emulsion composed of DOTAP and squalene was complexed with the plasmid DNA. An intravenous injection of cationic emulsion containing proinsulin gene decreased blood glucose levels for 7 days within normal range. The cationic emulsion exerted more profound effect on blood glucose levels compared to naked DNA. RT-PCR results confirmed that the proinsulin was expressed in several organs containing liver, lung, spleen, and kidney. The refractory response was invoked by multiple injections of naked DNA or cationic emulsion/DNA complex, which was later proven to be an immune response against expressed proinsulin. Therefore, the cationic emulsion showed a promising result as a novel insulin gene therapy vehicle by decreasing blood glucose level for a month.

Animals↗

Rip immunoreactivity significantly decreases in the stratum oriens of hippocampal CA1 region after transient forebrain ischemia in gerbils.

In the present study, we observed ischemia-related changes in Rip recognizing the promyelinating and myelinating oligodendrocytes in the hippocampus proper after 5 min of transient forebrain ischemia in gerbils. Rip immunoreactivity was significantly altered in the hippocampal CA1 region but not in the CA2/3 region after ischemic insult. In the sham-operated group, Rip immunoreactivity was shown in the cell bodies and processes of oligodendrocytes in all layers of the hippocampus proper. From 15 min to 2 days after ischemic insult, Rip immunoreactivity was similar to that of sham-operated group. Three days after ischemic insult, Rip-immunoreactive processes were tangled in the stratum oriens of the CA1 region, and Rip protein level decreased from this time after ischemia. Thereafter, Rip immunoreactivity was decreased time dependently in the CA1 region. Seven days after ischemic insult, Rip-immunoreactive processes were tangled and densely detected in the stratum oriens adjacent to the stratum pyramidale. In brief, these results indicate that the significant decrease of Rip immunoreactivity in processes in the stratum oriens of the hippocampal CA1 region occurs at late time after ischemia, and this decrease in Rip immunoreactivity may be associated with delayed neuronal death of CA1 pyramidal cells.

Animals↗

Transient ischemia-induced expression and changes of tyrosine kinase A in the hippocampal dentate gyrus of the gerbil.

The present study examined ischemia-related changes in tyrosine kinase A (trkA) immunoreactivity and its protein content in the dentate gyrus after 5 min of transient forebrain ischemia in gerbils. One day after ischemic insult, cresyl violet-positive polymorphic cells showed ischemic degeneration. The ischemia-induced changes in trkA immunoreactivity were found in the polymorphic layer (PL) and granule cell layer (GCL) of the dentate gyrus. In the sham-operated group, trkA immunoreactivity in the dentate gyrus was very weak. From 30 min after ischemia, trkA immunoreactivity was increased in the dentate gyrus and peaked in the dentate gyrus at 12 h after ischemia-reperfusion. Thereafter, trkA immunoreactivity was decreased time-dependently after ischemia-reperfusion. Four days after ischemic insult, trkA immunoreactivity was similar to that of the sham-operated group. In addition, it was found that ischemia-related changes in trkA protein content were similar to the immunohistochemical changes. These results suggest that the chronological changes of trkA in the dentate gyrus after transient forebrain ischemia may be associated with ischemic damage in polymorphic cells of the dentate gyrus.

Analysis of Variance↗

Berberry extract reduces neuronal damage and N-Methyl-D-aspartate receptor 1 immunoreactivity in the gerbil hippocampus after transient forebrain ischemia.

In the present study, we studied the neuroprotective effects of berberry extract (BE) against ischemic damage and the temporal and spatial alterations of N-methyl-D-aspartate receptor type 1 (NR1) and NR2A/2B immunoreactivities in the gerbil hippocampal CA1 region after transient ischemia to examine anti-ischemic effects and its role in transient forebrain ischemia. In the vehicle-treated group, the percentage of cresyl violet positive pyramidal cells in the CA1 region was about 11.4% compared to the sham-operated group 4 d after ischemic insult. BE showed neuroprotective effects against ischemic damage after ischemia-reperfusion. In the BE-treated groups, about 60-75% of CA1 pyramidal cells were stained with cresyl violet 4 d after ischemic insult. We observed the percentage of berberine (7.45+0.85 mg/g in BE) by HPLC, which is active ingredient of BE. NR1 immunoreactivity in the stratum pyramidale of the CA1 region in the vehicle-treated group was significantly increased at 30 min after transient forebrain ischemia, while at this time the NR1 immunoreactivity in the BE-treated groups was significantly low compared to the vehicle-treated group. The pattern of NR2A/B immunoreactivity in the stratum pyramidale of the BE-treated group and its protein levels were similar to that in the vehicle-treated group after ischemic insult. These results suggest that BE has potent neuroprotective effects against ischemic damage via the reduction of NR1 activity.

Animals↗

Release property of temperature-sensitive alginate beads containing poly(N-isopropylacrylamide).

The graft copolymer (APN) of alginate and poly(N-isopropylacrylamide) (PNIPAM) were synthesized and APN beads were prepared by dropping the aqueous solution of the copolymer into an aqueous solution of Ca(2+) solution. Alginate chains were employed to play a role in forming beads by electrostatic interactions with a multivalent ion, Ca(2+). Grafted PNIPAM segments were adopted to act as a valve for the pores of the beads, since they exhibit the properties of thermal contraction and expansion. The percent of release of blue dextran from APN beads was higher at 40 degrees C than at 25 degrees C. The difference in the release between two temperatures became more distinguishable when the content of PNIPAM in APN beads is higher. Below lower critical solution temperature (LCST), the expanded PNIPAM would close the pores of the beads, resulting in a lower release rate. Above LCST, the thermally contracted polymer would open the pores, resulting in a higher release rate. The percent of release from APN beads were investigated when the temperature of the release medium is altered. The release rate was relatively low at 25 degrees C. The temperature, however, changed up to 40 degrees C, a marked increase in the release rate was observed. These trends were found to be reproducible when the temperature was repeatedly altered between 25 and 40 degrees C. As a result, a stepwise response to the temperature alteration was obtained.

Acrylic Resins↗

Chronological changes of neurofilament 200 kDa immunoreactivity in the lateral olfactory tract after transient forebrain ischemia in gerbils.

This study was carried out to investigate the transient ischemia-induced changes of neurofilament 200 kDa (NF-200) immunoreactivity and protein content in the gerbil lateral olfactory tract (LOT) after 5 min of transient forebrain ischemia. Weak NF-200 immunoreactivity was detectable in the LOT in the sham-operated group. In this group, a few somata of mitral cells showed weak NF-200 immunoreactivity. One day after transient ischemia, NF-200 immunoreactivity in the LOT was increased significantly. NF-200 immunoreactivity in the LOT by 15 days after ischemia was similar to that in the 1 day post-ischemic group. In this time period, strong NF-200 immunoreactivity was expressed in the mitral cell processes, but the immunoreactivity in the mitral cell somata was significantly decreased. Thereafter, NF-200 immunoreactivity in the LOT was decreased significantly by 30 days after ischemic insult. At this time after ischemia, NF-200 immunoreactivity in the mitral cell dendrites was significantly decreased. The result of Western blot study showed that the pattern of NF-200 expression was similar to that of immunohistochemistry after ischemia-reperfusion. Our result suggests that changes of NF-200 protein in the gerbil LOT may be related to response to ischemic damage and that the axonal transport followed transient ischemia may be disturbed.

Animals↗

Ischemia-related changes in galanin expression in the dentate hilar region after transient forebrain ischemia in gerbils.

Although galanin (GAL) protects hippocampal neurons from ischemic damage, no study has examined ischemia-related changes in endogenous GAL in the hippocampal dentate gyrus. We investigated the chronological changes of GAL, well-known as the potassium channel opener, expression in the dentate gyrus at various times after 5 min of transient forebrain ischemia in gerbils. A few GAL-immunoreactive (IR) neurons were found in the polymorphic layer of the sham-operated group. Three hours after ischemia-reperfusion, the pattern of GAL immunoreactivity was similar to that of the sham-operated group and the number of GAL-IR neurons and immunoreactivity were highest 12 h after ischemic insult. At this time, GAL-IR neurons in the polymorphic layer showed strong GAL immunoreactivity. Thereafter, GAL-IR neurons and immunoreactivity significantly decreased in the dentate hilar region. Four days after ischemic insult, GAL-IR neurons were not detectable. In addition, the results of a Western blot study showed a pattern of GAL expression similar to the immunohistochemical changes. GAL protein content also was highest 12 h after ischemia. In conclusion, the increased expression of endogenous GAL in the dentate gyrus after ischemia is related to response to the ischemic damage.

Animals↗

Monoolein cubic phases containing hydrogen peroxide.

Monoolein (MO) cubic phases were prepared by hydrating MO using distilled water or 12wt.% H(2)O(2) solution so that the content of aqueous phase in the cubic phase is 30wt.%. The thermal transition of the isotropic cubic phase to reversed hexagonal phase was observed on a polarizing photomicroscope and the transition temperature was found to be around 65 degrees C on a differential scanning calorimeter (DSC). Small-angle X-ray scattering (SAXS) patterns indicated the cubic phases had diamond surfaces. The cubic phase released H(2)O(2) into an aqueous phase in a saturation manner so that approx. 50% of total loaded H(2)O(2) release in the first 10 h and thereafter relatively slow was observed over 40 h. The cubic phase was stable at 45 degrees C for 56 days before it broke down into an oily phase and an aqueous phase in 70 days. According to (1)H NMR spectrum, glycerol moiety and -CH(2)=CH(2)- of the oily phase were detected less in number than those of intact MO. Therefore, the hydrolysis and the oxidation of MO would be responsible for the breakdown of the cubic phase. The tensile adhesive forces of the cubic phases were higher than a skin-adhesive patch prepared using polyacrylate. The cubic phase containing H(2)O(2) could be used as a topical disinfected gel for a wounded skin.

Calorimetry, Differential Scanning↗

Two-dimensional packing patterns of amino acid surfactant and higher alcohols in an aqueous phase and their associated packing parameters.

An amino acid surfactant, monosodium N-stearoyl-L-glutamate (MSSG), was assumed to associate with higher alcohols (HAs) in 1:1 molar ratio in an aqueous phase, and the packing parameters of the 1/1 MSSG/HA associates, Pa, were calculated by molecular dynamics (MD). Pa is defined as At1(l(t1)+l(t2))/a(h)l(t1), where At1 stands for the cross-sectional area of the MSSG tail, l(t1) for the length of the MSSG tail, l(t2) for the length of the HA tail, and a(h) for the area of the MSSG head group. The Pa value increased from 0.83 to 1.02 as the HA tail length (l(t2)) increased from C14 to C22. Associates of HAs having longer tails with MSSG are likely to fit better into bilayers because Pa is closer to 1 when l(t2) is longer. In the graphical results of the MD simulation for the association, however, a steric hindrance was found between the head groups of MSSG/HA when l(t2) was > or = C19. Based on this result, HA was classified into short-chain HA (SCHA, l(t2) < C19) and long-chain HA (LCHA, l(t2) > or = C19), and several possible packing units, composed of compositional combinations of MSSG, SCHA, and LCHA, were proposed. The packing unit is a building block which could constitute bilayers, and it is composed of variable compositional combinations of MSSG/HA. Assuming that SCHA associates with MSSG, the packing parameter, Punit, was calculated in a water box by MD for each packing unit. Punit is the packing parameter of a packing unit and it is defined as Vu/l(u)a(u), where Vu is the tail volume of the packing unit, l(u) is the chain length, and a(u) is the head area. For the calculation, stearyl alcohol (C18-OH, SA) was chosen as a SCHA and behenyl alcohol (C22-OH, BA) as a LCHA. When the compositional ratio MSSG:SCHA:LCHA was 1:1:1, Punit was around 1. The packing unit having Punit of around 1 formed a colloidally stable suspension for 30 days and its aggregate was a lamellar structure. However, the other packing units, for which Punit deviates significantly from 1, precipitated out in their suspensions and showed no evidence of a lamellar structure. According to the graphical MD simulations for the compositional MSSG/SCHA/LCHA associations in bilayers, vertical steric hindrance was found between LCHAs when Punit deviated significantly from 1. The steric hindrance would prevent the packing units from forming a stable bilayer and induce precipitation in the suspensions. Therefore, a proper combinational ratio of MSSG:LCHA:SCHA would play a major role in forming a lamellar structure.

Alcohols↗

Age-related changes of parvalbumin immunoreactive neurons in the rat main olfactory bulb.

Parvalbumin (PV) is found in the olfactory system, including the main olfactory bulb, and is thought to be one of the neuroactive substances in olfaction. Changes in PV immunoreactivity in the olfactory system during aging have not been examined. We investigated such changes in the main olfactory bulb (MOB) of the rat at postnatal month 1 (PM 1), PM 3, PM 6, PM 12 and PM 24. PV-IR neurons were almost completely restricted to the external plexiform layer. At PM 1 there were only a few PV-IR neurons; at PM 3, the number of PV-IR neurons was at its greatest but they were not well developed morphologically. At PM 6, the number of PV-IR neurons was similar to that at PM 3 and they had satellite somata with well-developed processes with many varicosities. By PM 12 the number of neurons and processes had declined, and by PM 24, they had fallen even further and the remaining processes had lost most of their varicosities. We conclude that age-related degeneration of PV-IR neurons in the MOB may reduce calcium buffering and affect olfactory function in senile species.

Aging↗

Isolation and identification of an antioxidant enzyme catalase stimulatory compound from Garnoderma lucidum.

Antioxidant enzymes are scavenger reactive-oxygen intermediates and are involved in many cellular defense systems. We previously reported that a crude extract of Garnoderma lucidum, a medicinally potent mushroom, profoundly increased the catalase gene expression and enzyme activities in mouse livers (Park et al., J. Biochem. Mol. Biol. 34. 144-149, 2001). In this study, we elucidated the detailed mechanism whereby G. lucidum stimulates the catalase activity and expression. The major active fraction was isolated from G. lucidum and methyl linoleate was considered the most major component of the fraction. In order to determine whether methyl linoleate increases mRNA and protein synthesis of catalase, Northern and Western blot analyses were performed in vivo with methyl linoleate-treated mouse liver homogenate after feeding methyl linoleate to the mice. Northern and Western blot analyses of the crude liver homogenates in the mice that were administered methyl linoleate revealed that the expression catalase was significantly increased when compared to the untreated controls. In addition, the catalase protein levels and enzymatic activities increased in the mouse liver homogenates. These results suggest that methyl linoleate that is produced by G. lucidum stimulates the catalase expression at the transcription level.

Agaricales↗

Human liver catalase: cloning, expression and characterization of monoclonal antibodies.

We isolated a cDNA encoding liver catalase from a human liver cDNA library. The cDNA had a high degree of sequence similarity to the corresponding enzyme from other sources. It was expressed in E. coli using the pET15b vector. The protein produced was enzymatically active after purification, and its kinetic parameters closely resembled those of other mammalian catalases. Monoclonal antibodies were generated against the purified catalase; six antibodies recognizing different epitopes were obtained, one of which inhibited the enzyme. The cross reactions of the antibodies with brain catalases from human and other mammalian tissues were investigated, and all the immunoreactive bands obtained on Western blots had molecular masses of about 58 kDa. Similarly fractionated extracts of several mammalian cell lines all gave a single band of molecular mass 58 kDa. These results indicate that mammalian livers and human cell lines contain only one major type of immunologically reactive catalase, even though some of catalases have been previously reported to differ in certain properties.

Amino Acid Sequence↗

Gastrodin decreases immunoreactivities of gamma-aminobutyric acid shunt enzymes in the hippocampus of seizure-sensitive gerbils.

Gastrodin is one of the natural compound isolated from Gastrodia elata and has known anticonvulsant effects, although the exact pharmacological principles of this natural compound and its effects on other aspects of gamma-aminobutyric acid (GABA) metabolism in vivo have not been explored. Therefore, in the present study, the effects of gastrodin on GABA metabolism in the gerbil hippocampus were examined, in an effort to identify the antiepileptic characteristics of this substance. Gastrodin reduced the seizure score in the treated group, although the immunoreactivities of GABA synthetic enzymes and GABA transporters were unaltered in gastrodin-treated animals. Interestingly, in the gastrodin-treated group, GABA transaminase (GABA-T) immunoreactivity in the hippocampus, particularly in neurons, was significantly decreased. In the gastrodin-treated group, both succinic semialdehyde dehydrogenase (SSADH) and succinic semialdehyde reductase (SSAR) immunoreactivities in the hippocampus was also decreased significantly, which stood in contrast to the nontreated group, in which strong SSADH and SSAR immunoreactivities were detected. From the neuroanatomical viewpoint, these findings suggest that gastrodin may cause the elevation of GABA concentration by inhibiting the GABA shunt.

Animals↗

The altered expression of GABA shunt enzymes in the gerbil hippocampus before and after seizure generation.

In the present study, the distribution of succinic semialdehyde dehydrogenase (SSADH) and succinic semialdehyde reductase (SSAR) in the hippocampus of the Mongolian gerbil and its association with various sequelae of spontaneous seizure were investigated in order to identify the roles of GABA shunt in the epileptogenesis and the recovery mechanisms in these animals. Both SSADH and SSAR immunoreactivities in the GABAergic neurons were significantly higher in the pre-seizure groups of seizure sensitive (SS) gerbil as compared to those seen in the seizure resistant (SR) gerbils. The distributions of both SSADH and SSAR immunoreactivities in the hippocampus showed significant differences after the on-set of seizure. At 3 h postictal, when compared to the pre-seizure group of SS gerbils, a decline in the immunoreactivities in the perikarya was observed. At 12 h after seizure on-set, the densities of both SSADH and SSAR immunoreactivities were begun to recover to the pre-seizure level of SS gerbils. These results suggest that the GABAergic neurons in the hippocampal complex of the SS gerbil may be highly activated. In addition, the imbalance of GABA shunt expressions in the GABAergic neurons may imply a malfunction of the metabolism of GABAergic neurons in the SS gerbils, and this defect may trigger seizure on-set. Therefore, the initiation of seizure, at least in gerbils, may be the result of a malfunction in GABA shunt in the GABAergic neurons.

Aldehyde Oxidoreductases↗

Effect of the flavonoid components obtained from Scutellaria radix on the histamine, immunoglobulin E and lipid peroxidation of spleen lymphocytes of Sprague-Dawley rats.

The effect on the IgE content induced by concanavalin A in spleen lymphocytes of the presence wogonin, ganhuangenin, wogonoside and 3,5,7,2',6'-pentahydroxyl flavanone was investigated. These flavonoid components markedly inhibited the histamine released from cells stimulated with the calcium ionophore, A23187. However, the magnitude of the inhibitory effect on the degree of lipid peroxidation by ConA of these components was in order of PHF>GHG>WG>WGS. Interestingly, WG, GHG and WGS, with a methoxyl group in the A and B rings, strongly inhibited histamine and IgE production, whereas PHF without a methoxyl group was much stronger than that for lipid peroxidation. We suggest that WG, GHG and WGS might block the pathway for the release of histamine, and that the IgE level in spleen lymphocytes is responsible for the lipid peroxidation.

Animals↗

Efficacy of dietary aloe vera supplementation on hepatic cholesterol and oxidative status in aged rats.

In the current study, we show the anti-oxidative and hypocholesterol effects of aloe vera in the liver. Male specific pathogen-free (SPF) Fischer 344 rats were randomly assigned to one of four groups: Group A (control) was fed test chow without aloe supplementation; Group B was fed a diet containing a 1% (per weight basis) freeze-dried aloe filet; Group C was fed a diet containing a 1% (per weight basis) charcoal-processed, freeze-dried aloe filet; and Group D was fed a diet containing a charcoal-processed freeze-dried, whole leaf aloe (0.02% per weight basis) in the drinking water. Our results show that a life-long intake of aloe had superior anti-oxidative action against lipid peroxidation in vivo, as indicated by reduced levels of hepatic phosphatidylcholine hydroperoxide. Additional anti-oxidative action was evidenced by enhanced superoxide dismutase (SOD) and catalase activity in groups B and C. Furthermore, our study revealed that hepatic cholesterol significantly increased in the control group during aging in contrast to the aloe-supplemented groups, which showed approximately 30% lower cholesterol levels, thereby an effective hypocholesteremic efficacy. In this report, we suggest that life-long dietary aloe supplementation suppresses free radical-induced oxidative damage and age-related increases in hepatic cholesterol.

Aging↗

Changes in pyridoxal kinase immunoreactivity in the gerbil hippocampus following spontaneous seizure.

To identify the roles of pyridoxal kinase (PLK) in epileptogenesis and the recovery mechanisms in spontaneous seizure, a chronological and comparative analysis of PLK expression in the gerbil hippocampus was conducted. PLK immunoreactivity in a pre-seizure group of seizure sensitive (SS) gerbils was more strongly detected than that in a seizure resistant (SR) group. The density of PLK immunoreactivity in a 30-min postictal group was significantly lower than that of a pre-seizure group. In a 12 h postictal group, PLK immunodensity recovered to pre-seizure level. The over-expression of PLK in the hippocampus of pre-seizure SS gerbils suggests that PLP play an important role in the modulation of GAD activity and GABA reuptake as mediated by membrane transporter via neurons.

Animals↗