• Journal of Microbiology and Biotechnology
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• v.25 no.9
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• pp.1485-1492
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• 2015

Makgeolli is a traditional wine in Korea and has been traditionally believed to exhibit health benefits. However, the inhibitory effect of dealcoholized makgeolli (MK) on cancer has never been investigated scientifically. In this study, MK exhibited an anti-angiogenic effect by inhibiting tube formation in human umbilical vein endothelial cells, without cytotoxicity. Treatment with MK reduced the proliferation of AGS human gastric adenocarcinoma cells in a dose-dependent manner and increased the sub-G1 population. Next, we evaluated whether MK could induce apoptosis in AGS cells by using the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay or Annexin V method. Treatment with MK at 500 and 1,000 μg/ml increased the number of TUNEL-positive AGS cells. Under the same conditions, MK-treated (500 and 1,000 μg/ml) cells showed significant induction of early or late apoptosis, compared with untreated cells (no induction). In addition, MK also induced phosphatase and tensin homolog (PTEN) expression in AGS cells. However, p53 expression in AGS cells was not changed by MK treatment. Furthermore, MK at 500 mg/kg·d reduced the tumor size and volume in AGS tumor xenografts. Taken together, MK may be useful for the prevention of cancer cell growth.

• YAKHAK HOEJI
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• v.40 no.4
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• pp.422-427
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• 1996

The anti-Ex-A IgG was specifically labeled with stable isotopes, DL-His-2,4-$d_2$, L-Phe-$d_5$, L-Trp-$d_5$, L-Tyr-2,6-$d_2$ and L-[1-$^{13}C$]Trp, by growing hybridoma cell in serum-free medium. By use of NMR spectroscopy with selectively labeled Fab fragment, we applied a paratope mapping on antigen-antibody complex. Assignments of the observed carbonyl carbon resonances have been determined by using $^{13}C$-$^{15}N$ double labeling method in order to assign the Trp resonances. Photo CIDNP was also applied to investigate the antigen-binding site(s) on the surface residues of antibody. We found that Trp 36, which is located at the $V_H$ domain, is an important residue to bind to Ex-A, however, two Tyr on the surface of anti-Ex-A IgG plays no crucial role to bind to antigen. On the basis of these results, we demonstrate that stable isotope-aided NMR strategy can be extended to molecular structural analyses of the complex of an Fab fragment and a protein antigen.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.3
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• pp.1-13
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• 2009

Objectives : This study evaluates neuroprotective effect of Gastrodiae Rhizoma on apoptosis in the cerebral infarct. Methods : Cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method in Sprague-Dawley rats. Then ethanol extract of Gastrodiae Rhizoma was administered orally for 3 days. Infarct area and volume were evaluated with TTC staining. Neuronal apoptosis was identified with TUNEL labeling. Apoptosis modulatory effect was observed with immunohistochemical Bax, Bcl-2, iNOS, and MMP-9 expressions in penumbra. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced infarct size partly and volume significantly of in the MCAO rat brain. 2. Ethanol extract of Gastrodiae Rhizoma reduced TUNEL positive cell ratio in the penumbra of MCAO rat brain significantly. 3. Ethanol extract of Gastrodiae Rhizoma suppressed Bax, iNOS and MMP-9 expression in the penumbra of MCAO rat brain significantly. 4. Ethanol extract of Gastrodiae Rhizoma did not change Bcl-2 expression in the penumbra of MCAO rat brain. But expression ratio of Bcl-2 against Bax was increased in the Gastrodiae Rhizoma group. Conclusions : These results suggest that Gastrodiae Rhizoma plays an anti-apoptotic neuroprotective effect through suppression of Bax, iNOS, and MMP-9 expressions and relative up-regulation of Bcl-2 in the ischemic brain tissue.

• Journal of Biosystems Engineering
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• v.39 no.2
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• pp.111-114
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• 2014

Purpose: This review aims to introduce aptamers and the methods of its development to improve the sensitivity and selectivity to target bacteria. In this review, we have highlighted current developments and directions in the pathogen detection based on aptamers. Background: Aptamers, the specific nucleic acid sequences, can bind to targets with high affinity and specificity. Some of researches on the use of aptamers for the detection of pathogen have been reported in recent years. Aptamers have more applicability than antibodies for the development of pathogen detection using biosensor; such as easy to synthesis and labeling, lack of immunogenicity, and a low cost of production. However, only few reports on the development and use of aptamers for the detection of pathogen have been published. Review: Aptamers specific to pathogen are obtained by whole-cell systematic evolution of ligands by exponential enrichment (SELEX) process. SELEX process is composed of screening random oligonucleotide bound with target cells, multiple separation and amplification of nucleic acids, final identification of the best sequences. For improving those affinity and selectivity to target bacteria, optimization of multiple separating process to remove unbounded oligonucleotides from aptamer candidates and sorting process by flow cytometry are required.

• Proceedings of the Korea Crystallographic Association Conference
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• 2002.11a
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• pp.54-55
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• 2002

The fabrication, characterization and manipulation of nanoparticle system brings together physics, chemistry, materials science and biology in an unprecedented way. Phenomena occurring in such systems are fundamental to the workings of electronic devices, but also to living organisms. The ability to fabricate the surface of nanoparticles Is essential in the further development of functional devices that incorporate nanoscale features. Even more essential is the ability to introduce a wide range of chemical and materials flexibility into these structures to build up more complex nanostructures that can ultimately rival biological nanosystems. In this respect, polymers are potentially ideal nanoscale building blocks because of their length scale, well-defined architecture, controlled synthesis, ease of processing and wide range of chemical functionality that can be incorporated. In this presentation, we will look at a number of promising polymer-based nanoparticle fabrication strategies that have been developed recently, with an emphasis on those techniques that incorporate nanostructured polymeric particles into electronic devices or biomedical applications. And functional nanoparticles deliberately designed using several powerful process methods and their application will be discussed. Nanostructured nanoparticles, what we called, implies dispersed colloids with the size ranged from several nanometers to hundreds of nanometer. They have extremely large surface area, thus it is very important to control the morphology or surface functionality fitted for adequate objectives and properties. Their properties should be controlled for various kind of bio-related technologies, such as immunomagnetic cell separation, drug delivery systems, labeling and identification of lymphocyte populations, extracorporeal and hemoperfusion systems, etc. Well-defined polymeric nanoparticles can be considered as smart bomb or MEMS.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.566-571
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• 2017

Background: A number of reports have described the protective effects of ginsenoside Rg1 (Rg1) in Alzheimer's disease (AD). However, the protective mechanisms of Rg1 in AD remain elusive. Methods: To investigate the potential mechanisms of Rg1 in ${\beta}$-amyloid peptide-treated SH-SY5Y cells, a comparative proteomic analysis was performed using stable isotope labeling with amino acids in cell culture combined with nano-LC-MS/MS. Results: We identified a total of 1,149 proteins in three independent experiments. Forty-nine proteins were significantly altered by Rg1 after exposure of the cells to ${\beta}$-amyloid peptides. The protein interaction network analysis showed that these altered proteins were clustered in ribosomal proteins, mitochondria, the actin cytoskeleton, and splicing proteins. Among these proteins, mitochondrial proteins containing HSD17B10, AARS2, TOMM40, VDAC1, COX5A, and NDUFA4 were associated with mitochondrial dysfunction in the pathogenesis of AD. Conclusion: Our results suggest that mitochondrial proteins may be related to the protective mechanisms of Rg1 in AD.

• Biomedical Science Letters
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• v.9 no.2
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• pp.67-74
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• 2003

Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.226-229
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• 2012

Bacterial hemoglobin from Vitreoscilla (VHb) is recognized as a good fusion protein for the soluble expression of foreign protein. In this study, we generated a monoclonal antibody (MAb) against VHb for its detection. For the rapid screening of MAb, a protein chip technology based on the Alexa-488 (A488) dye labeling method was introduced. In order to fabricate the chip, the VHb protein was chemically coupled to the chip surface and then the culture supernatants of 84 hybridoma cell lines were spotted onto the VHb chip. The bound MAbs were measured by A488-modified anti-mouse IgG. A single spot (MAb A10) exhibited significantly high signal intensity. The immunoblot analysis evidenced that the MAb A10 can detect VHb-fused proteins with high specificity.

• Journal of Korean Neurosurgical Society
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• v.39 no.6
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• pp.432-437
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• 2006

Objective : In order to establish the index of degeneration, the authors performed a histochemical study with Safranin-O staining and investigated the occurrence of apoptosis in the human intervertebral disc. Methods : Eighteen intervertebral disc specimens surgically extracted from the patients and two additional specimens from the autopsied cases were stained with Safranin-O for proteoglycan according to a standard protocol. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate- biotin nick end labeling[TUNEL] was used to detect the fragmented DNA known to be associated with apoptotic cell death and classification scheme was formulated for categorization of the degree of Safranin-O staining [normal, moderate reduction, faint] by modification of Makin's histological-histochemical grading. The Kruskal-Wallis H test and Chi-square test were used for statistical analysis. Results : The statistical results showed a significant difference in the mean age between "normal" Safranin-O staining group and the others [19.3 versus 55, 43.4, p=0021]. However, there was no statistically significant correlation between Safranin-O staining and MR grading of disc degeneration. Only six of eighteen surgical specimens and none in autopsies showed positive apoptotic cells in TUNEL staining. Conclusion : The determination of the degree of degeneration in surgically obtained disc tissue per se by histochemical staining or by the degree of apoptosis that corresponds to its morphologic change was not feasible.

• Herbal Formula Science
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• v.31 no.3
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• pp.183-191
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• 2023

Objective : UVB damages skin health by causing skin redness and intense inflammation, sunburn, and skin cancer. Paeoniae Radix Alba has been used to relieve gynecological symptoms, muscle spasms, and skin ailments. This study was conducted to confirm whether it has a protective effect against UVB photodamage. Methods : Ethanol extract of Paeoniae Radix Alba (PRA) was prepared by extracting 100 g Paeoniae Radix Alba in 1 L of ethanol for 48 h. Apoptosis was monitored by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and expression levels of apoptosis indicator proteins, and tyrosinase activity was measured with a colorimetric commercial kit. Results : In human keratinocyte HaCaT cells, PRA reduced UVB-induced cell death through apoptosis by inhibiting PARP cleavage and caspase-3 and -9. UVB-induced increase in cellular reactive oxygen species (ROS) was suppressed by PRA pretreatment. PRA also showed dose-dependent ABTS and DPPH radical scavenging activities. Furthermore, the inhibitory effect of tyrosinase activity by PRA was confirmed. Conclusion : These results demonstrated the protective role of PRA in UVB photodamage of human keratinocytes, mainly due to its antioxidant and antiapoptotic properties. We also suggest that PRA can be considered as an effective natural agent to prevent skin photodamage.