• The Korean Journal of Physiology
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• v.30 no.2
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• pp.197-208
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• 1996

Endosomes lower their internal pH by an ATP-driven proton pump, which is critical to dissociation of many receptor-ligand complexes, the first step in the intracellular sorting of internalized receptors and ligands. Endosomes are known to exhibit n great range of pH values that can vary between 5.0 and 7.0 within a single cell although the factors that regulate endosomal pH remain uncertain. To evaluate the morphological and topological differences of endosomes in the different stages, confocal microscopy was used. The early endosomes labeled with fluorescein isothiocyanate-dextran for 10 min at $37^{\circ}C$ were identifiable at the peripheral and tubule-vesicular endosome compartment. In contrast, the late endosomes formed by 10 min pulse and 20 min trace were located deeper in the cytoplasm and showed more vesicular features than early endosomes. For the purpose of determining whether ATP-dependent acidification was heterogeneous and whether the differences in acidification were attributed to differences in the activity of $Na^{+}-K^{+}$-ATPase and/or $Cl^{-}$ channel, endocytic compartments were fractionated into subpopulation using percoll gradient and measured ATP-dependent acidification. While all fractions exhibited ATP-dependent acidification activity, both the initial rate of acidification and extent of proton translocation were lower in early endosomes and gradually increased in late endosomes. Phosphorylation by PKA and ATP enhanced ATP-dependent acidification in both early and late endosomes, hut there was no difference in the degree of enhancement by phosphorylation between two subpopulations. When ATP-dependent acidification was determined in the presence or absence of vanadate ($Na_{3}VO_{4}$) or ouabain, only early endosomes exhibited the vanadate or ouabain dependent stimulation of acidification activity, suggesting the inhibition of $Na^{+}-K^{+}$-ATPase. Therefore, it seems probable that the inhibition of early endosome acidification by $Na^{+}-K^{+}$-ATPase observed in vitro at least in part plays a physiological role in controlling the acidification of early endosomes in vivo.

• Korean Journal of Materials Research
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• v.22 no.2
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• pp.66-70
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• 2012

Ti-Ni alloys are widely used in numerous biomedical applications (e.g., orthodontics, cardiovascular science, orthopaedics) due to their distinctive thermomechanical and mechanical properties, such as the shape memory effect, superelasticity and low elastic modulus. In order to increase the biocompatibility of Ti-Ni alloys, many surface modification techniques, such as the sol-gel technique, plasma immersion ion implantation (PIII), laser surface melting, plasma spraying, and chemical vapor deposition, have been employed. In this study, a Ti-49.5Ni (at%) alloy was electrochemically etched in 1M $H_2SO_4$+ X (1.5, 2.0, 2.5) wt% HF electrolytes to modify the surface morphology. The morphology, element distribution, crystal structure, roughness and energy of the surface were investigated by scanning electron microscopy (SEM), energy-dispersive Xray spectrometry (EDS), X-ray diffractometry (XRD), atomic force microscopy (AFM) and contact angle analysis. Micro-sized pores were formed on the Ti-49.5Ni (at%) alloy surface by electrochemical etching with 1M $H_2SO_4$+ X (1.5, 2.0, 2.5) wt% HF. The volume fractions of the pores were increased by increasing the concentration of the HF electrolytes. Depending on the HF concentration, different pore sizes, heights, surface roughness levels, and surface energy levels were obtained. To investigate the osteoblast adhesion of the electrochemically etched Ti-49.5Ni (at%) alloy, a MTT test was performed. The degree of osteoblast adhesion was increased at a high concentration of HF-treated surface structures.

• BMB Reports
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• v.32 no.6
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• pp.585-593
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• 1999

The water and methanol extracts of Artemisia argyi showed significant cytotoxicities against J774A.1 cells but not so much against normal leukocytes. The cytotoxicities were found to be dependent on the extract concentration and the incubation time. The concentration of water and methanol extracts inhibiting 50% of cell proliferation ($IC_{50}$) were estimated to be 44.2 mg/ml and 71.6 mg/ml, respectively. In the presence of Artemisia argyi water extract, total superoxide dismutase (CuZnSOD and MnSOD) activities of media, cytoplasmic and mitochondrial fractions of J774A.1 cells increased in accordance with cytotoxicity. MnSOD was found to be the main component of enhanced total SOD activities, particulary in the mitochondrial fraction. In contrast to SOD, catalase and glutathione peroxidase (GPx) were not found in any instance of the current investigation. In addition, substantial amount of $O_2^-$ appeared to be generated in the mitochondrial fraction under the influence of Artemisia argyi. All data put together, it is postulated that Artemisia argyi extracts seem to stimulate $O_2^-$ generation in mitochondria of J774A.1 cells with concomitant increases of SODs. Since $H_2O_2$, the reaction product of SOD on $O_2^-$, is known to be readily converted to very toxic $OH{\cdot}$ in the absence of catalase and/or GPx cooperation, toxicity derived from ROS such as $O_2^-$, $H_2O_2$, and $OH{\cdot}$ may be the main cause of necrosis and/or apoptosis of J774A.1 cells.

• Journal of Korean Neurosurgical Society
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• v.46 no.6
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• pp.538-544
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• 2009

Objective : Primary treatment of spinal metastasis has been external beam radiotherapy. Recent advance of technology enables radiosurgery to be extended to extracranial lesions. The purpose of this study was to determine the clinical effectiveness and safety of stereotactic radiosurgery using Cyberknife in spinal metastasis. Methods : From June, 2002 to December, 2007, 129 patients with 167 spinal metastases were treated with Cyberknife. Most of the patients (94%) presented with pain and nine patients suffered from motor deficits. Twelve patients were asymptomatic. Fifty-three patients (32%) had previous radiation therapy. Using Cyberknife, 16-39 Gy in 1-5 fractions were delivered to spinal metastatic lesions. Radiation dose was not different regarding the tumor pathology or tumor volume. Results : After six months follow-up, patient evaluation was possible in 108 lesions. Among them, significant pain relief was seen in 98 lesions (91%). Radiological data were obtained in 83 lesions. The mass size was decreased or stable in 75 lesions and increased in eight lesions. Radiological control failure cases were hepatocellular carcinoma (5 cases), lung cancer (1 case), breast cancer (1 case) and renal cell carcinoma (1 case). Treatment-related radiation injury was not detected. Conclusion : Cyberknife radiosurgery is clinically effective and safe for spinal metastases. It is true even in previously irradiated patients. Compared to conventional radiation therapy, Cyberknife shows higher pain control rate and its treatment process is more convenient for patients. Thus, it can be regarded as a primary treatment modality for spinal metastases.

• Journal of Applied Biological Chemistry
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• v.58 no.2
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• pp.125-129
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• 2015

In this study, the antioxidant and anti-wrinkling effects of fraction from Vitex trifolia L. were investigated. Among the fractions, ethyl acetate fraction showed the highest antioxidant activities in 1-1-diphenyl-2-picryl-hydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoine-6-sulfonic acid) radical scavenging and elastase inhibition with 76, 89, and 74%, respectively, at a concentration of $1,000{\mu}g/mL$. This fraction, at the concentration of $25{\mu}g/mL$, inhibited 70% fibroblast cell viability and 86% the matrix metalloprotease (MMP)-1. In addition, the results from Western blot assay showed that this fraction ($25{\mu}g/mL$) expressed the MMP-1 protein level by decreasing 50%. The findings suggest that the ethyl acetate fraction from V. trifolia has great potential as a cosmeceutical ingredient with antioxidant and anti-wrinkling effects.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.777-783
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• 2004

Samples of 13 species of forest tree leaves fed to livestock in the semi-hilly arid zone of Punjab State in India were collected at 30 d interval for 12 months, in order to assess their nutritional worth for livestock. The ground samples were pooled for 4 different seasons viz. dry hot, hot humid, fall and winter. The chemical composition irrespective of the season revealed that CP content varied between 8.9 (Carrisa) to 22.0% (Leucaena). Globulin was the major protein fraction in most of the leaves. The lowest concentration of cell wall constituents was observed in Morus alba and Grewea. The leaves in general became fiberous and lignified during winter and fall as compared to summer season. The leaves of Grewea, Morus alba, Leucaena, Carrisa and Acacia were rich in Ca, P and most of the trace elements. The total phenolics ranged between 1.88% (Azardirachta) to 15.82% (Acacia). The leaves of Acacia had the highest concentration of hydrolysable tannins (14.6%) whereas that of Carrisa had that of condensed tannins (5.9%). The condensed tannins (more than 3%) were negatively correlated to the digestibility of dry matter (DM), neutral detergent fiber (NDF) and crude protein (CP). The digestion kinetic parameters for DM, NDF and CP revealed that leaves of Morus alba, Zizyphus and Ehretia had highest insoluble but potentially degradable fraction. The minimum rumen fill values also revealed that leaves of Grewea, Azardirachta, Morus, Ehretia and Leucaena had great potential for voluntary DM intake. The leaves of Ougeinia, Malha, Dodenia and Carrisa had significantly higher rumen fill value indicating poor potential for voluntary DM intake. Season did not have any significant impact on digestion kinetic parameters except that most of the leaves had low potentially degradable fraction, which was degraded at slow rate during winter. It was concluded that the leaves of Morus, Ehretia, Grewea and Leucaena had great potential as livestock feed, while feeding of Ougeinia, Malha and Dodonea leaves should be avoided.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1368-1376
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• 2008

In our previous study, the expression of active H-ferritins in Saccharomyces cerevisiae was found to reduce cell growth and reactive oxygen species (ROS) generation upon exposure to oxidative stress; such expression enhanced that of high-affinity iron transport genes (FET3 and FTR1). The results suggested that the recombinant cells expressing H-ferritins induced cytosolic iron depletion. The present study analyzes metabolic changes under these circumstances via proteomic methods. The YGH2 yeast strain expressing A-ferritin, the YGH2-KG (E62K and H65G) mutant strain, and the YGT control strain were used. Comparative proteomic analysis showed that the synthesis of 34 proteins was at least stimulated in YGH2, whereas the other 37 proteins were repressed. Among these, the 31 major protein spots were analyzed via nano-LC/MS/MS. The increased proteins included major heat-shock proteins and proteins related to endoplasmic reticulum-associated degradation (ERAD). On the other hand, the proteins involved with folate metabolism, purine and methionine biosynthesis, and translation were reduced. In addition, we analyzed the insoluble protein fractions and identified the fragments of Idh1p and Pgk1p, as well as several ribosomal assembly-related proteins. This suggests that intracellular iron depletion induces imperfect translation of proteins. Although the proteins identified above result from changes in iron metabolism (i.e., iron deficiency), definitive evidence for iron-related proteins remains insufficient. Nevertheless, this study is the first to present a molecular model for iron deficiency, and the results may provide valuable information on the regulatory network of iron metabolism.

• Fisheries and Aquatic Sciences
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• v.15 no.2
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• pp.137-143
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• 2012

The abalone Haliotis discus hannai, is one of the economically important species in the fisheries industry. Abalone intestines are one of the by-products of its processing. To investigate its bioactive potential, abalone intestine was digested using an in vitro gastrointestinal (GI) digestion system containing pepsin, trypsin, and ${\alpha}$-chymotrypsin. The abalone intestine G1 digests (AIGIDs) produced by the GI digestion system were fractionated into AIGID I (> 100 kDa), AIGID II (10-100 kDa), and AIGID III (1-10 kDa) using an ultrafiltration membrane system. Of the three digests, AIGID II and AIGID III exhibited inhibitory effects against matrix metalloproteinase-2 and -9 (MMP-2, MMP-9) in HT1080 human fibrosarcoma cells. Both fractions potently inhibited gelatine digestion by MMP-2 and MMP-9 treated with phorbol 12-myristate 13-acetate (PMA) and migration of HT1080 cells in dose dependently. Furthermore, AIGID II and III attenuated expression of p65, a component of nuclear transcription factor kappa B. These results indicate that of the abalone intestine digests inhibit MMP-2 and MMP-9. Thus, the AIGIDs or their active components may have preventive and therapeutic potential for diseases associated with MMP-2 and MMP-9 activation in fibrosarcoma cells.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.10
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• pp.1498-1504
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• 2005

Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

• Journal of Microbiology and Biotechnology
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• v.27 no.6
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• pp.1090-1097
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• 2017

Rhus verniciflua Stokes (RVS), an herbal medicine found in East Asia, was extracted and further fractionated to investigate its antioxidant capacity and neuroprotective effects. The RVS ethyl acetate (EtOAc) fraction had the highest level of total phenolics and antioxidant capacity among all solvent fractions tested. Pretreatment of PC-12 cells with the EtOAc fraction effectively attenuated $H_2O_2$-induced oxidative damage. Furthermore, the EtOAc fraction significantly attenuated caspase-3 activity, resulting in inhibition of $H_2O_2$-induced apoptosis. We identified and quantified fustin, sulfuretin, and butein in the EtOAc fraction using accurate mass quadrupole time-of-flight mass spectrometry and reversed-phase high-performance liquid chromatography. The intracellular antioxidant capacity and superoxide dismutase (SOD) activity were significantly increased in PC-12 cells treated with the EtOAc fraction and with individual flavonoids. When cells were pretreated with the EtOAc fraction or individual flavonoids and then co-incubated with diethyldithiocarbamic acid (an inhibitor of SOD activity), cell viability against $H_2O_2$-induced oxidative stress was attenuated. These results suggest that the RVS EtOAc fraction and its flavonoid constituents protect PC-12 cells against $H_2O_2$-induced neurotoxicity through their antioxidant properties.