• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1258-1266
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• 2005

The present study was carried out in order to assess the protective effects of calycosin-7-O-$\beta$-D-glucopyranoside, isolated from Astragali radix (AR), on hyaluronidase (HAase) and the recombinant human interleukin-$1\beta$ (IL-$1\beta$)-induced matrix degradation in human articular cartilage and chondrocytes. We isolated the active component from the n-butanol soluble fraction of AR (ARBu) as the HAase inhibitor and structurally identified as calycosin-7-O-$\beta$-D-glucopyranoside by LC-MS, IR, ${1}^H$ NMR, and ${13}^C$ NMR analyses. The $IC_{50}$ of this component on HAase was found to be 3.7 mg/ml by in vitro agarose plate assay. The protective effect of ARBu on the matrix gene expression of immortalized chondrocyte cell line C28/I2 treated with HAase was investigated using a reverse transcription polymerase chain reaction (RT-PCR), and its effect on HAase and IL-$1\beta$-induced matrix degradation in human articular cartilage was determined by a staining method and calculating the amount of degraded glycosaminoglycan (GAG) from the cultured media. Pretreatment with calycosin-7-O-$\beta$-D-glucopyranoside effectively protected human chondrocytes and articular cartilage from matrix degradation. Therefore, calycosin-7-O-$\beta$-D-glucopyranoside from AR appears to be a potential natural ant-inflammatory or antii-osteoarthritis agent and can be effectively used to protect from proteoglycan (PG) degradation.

• Journal of Microbiology and Biotechnology
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• 제27권8호
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• pp.1519-1528
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• 2017

Foodborne contamination and salmonellosis caused by Salmonella Enteritidis (S. Enteritidis) are a significant threat to human health and poultry enterprises. Outer membrane vesicles (OMVs), which are naturally secreted by gram-negative bacteria, could be a good vaccine option because they have many biologically active substances, including lipopolysaccharides (LPS), outer membrane proteins (OMPs), and phospholipids, as well as periplasmic components. In the present study, we purified OMVs derived from S. Enteritidis and analyzed their characteristics through silver staining and sodium dodecyl sulfate polyacrylamide gel electrophoresis. In total, 108 proteins were identified in S. Enteritidis OMVs through liquid chromatography tandem mass spectrometry analysis, and OMPs, periplasmic proteins, and extracellular proteins (49.9% of total proteins) were found to be enriched in the OMVs compared with bacterial cells. Furthermore, native OMVs used in immunizations by either the intranasal route or the intraperitoneal route could elicit significant humoral and mucosal immune responses and provide strong protective efficiency against a lethal dose (~100-fold $LD_{50}$) of the wild-type S. Enteritidis infection. These results indicated that S. Enteritidis OMVs might be an ideal vaccine strategy for preventing S. Enteritidis diseases.

• 대한치과교정학회지
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• 제11권2호
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• pp.161-168
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• 1981

This study was undertaken to observe the histologic changes of surrounding maxillary sutures to the widening of midpalatal suture, using two adult dogs, weighing about 10 kg, for experimental, and one for control group. After widening of the midpalatal suture with expansion screw for ten days was performed, and followed by sacrifice of experimental animals. Tissues were excised from 5 surrounding maxillary suture portions such as, internasal, interfrontal, midsagittal, zygomatrco-temporal, and midpalatal sutures. After that, the specimens were fixed and decalcified in $10\%$ formalin sol. and $5\%$ nitric acid. Embedding in paraffin and serial sections at a thickness of 5 micron was done, After Hematoxylin-Eosin staining and light microscopic examination, Following results were obtained: 1. In midpalatal suture area, which showed most prominent histologic change, High degree of fibronlastic and osteoblastic proliferation lining the bony trabeculae with osteoid tissue projection into the fibrous connective tissue were observed. 2. In interfrontal suture area, moderate degree of fibroblastic and osteoblastic proliferation was observed. 3. In internasal suture area, active osteoblastic and osteoclastic proliferation lining the bony trabeculae was observed, and separation of fibrous connective tissue was also observed. 4. In midsagittal and zygomatico-temporal suture areas, no histologic changes can be observed.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.563-566
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• 2000

키토산으로부터 키토산 올리고당을 생산하기 위한 유용 효소원을 개발하기 위해서 토양에서 4종의 미생물을 분리하였으며, 가장 높은 활성의 chitosanase를 생산하는 Bacillus sp. HSB-21 균주를 선정하였다. 분리균주로부터 생산된 chitosanase는 endo-type의 효소로 분자량이 약 21,000이었다. 최적 pH와 온도는 각각 5.5, $50^{\circ}C$이었으며, pH $3{\sim}8$ 범위와 $40^{\circ}C$까지 비교적 안정한 효소로 나타났다. 본 효소는 키토산을 분해하여 단당을 형성하지 않았으며 주로 $2{\sim}8$당을 포함하는 키토산 올리고당을 만들었다. 그러므로 본 효소는 고중합도 키토산 올리고당의 생산을 위한 응용 가능성이 큰 것으로 판단된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권1호
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• pp.183-187
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• 2016

Rhizomes of Boesenbergia pandurata (Roxb.) Schlecht have been reported to contain active compounds with anticancer properties. This research was carried out to examine anti-proliferative and apoptotic induction against HeLa and Vero cells-line. Dried powder of B. pandurata rhizomes was extracted by a maceration method using 90% ethanol. Cytotoxic assays to determine $IC_{50}$ and anti-proliferative effects were carried out by MTT methods. Observation of apoptosis was achieved with double staining using acridine orange and ethidium bromide. The results showed that ethanolic extract of B. pandurata was more cytotoxic against HeLa cells ($IC_{50}$ of $60{\mu}g/mL$) than Vero cells ($IC_{50}$ of $125{\mu}g/mL$). The extract had higher anti-proliferative activity as well as apoptotic induction in HeLa than Vero cells. Therefore, it was concluded that the ethanolic extract of B. pandurata had anti-proliferative as well as apoptosis induction activity dependent on the cell type.

• Asian Pacific Journal of Cancer Prevention
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• 제15권2호
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• pp.797-802
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• 2014

We screened a small molecular library that was designed and independently synthesized in vitro and found a new drug (MY-03-01) that is active against ovarian cancer. We established that MY-03-01 effectively inhibited SKOV-3 cell survival in a dose-dependent manner, based on cell viability rates, and that it not only induced SKOV-3 apoptosis by itself, but also did so synergistically with paclitaxel. Secondly, when MY-03-01 was applied at $40{\mu}M$, its hemolytic activity was less than 10%, compared with the control, and there was almost no damage to nor mal cells at this concentration. In addition, we used DAPI staining and flow cytometry to show that MY-03-01 could significantly induce apoptosis of SKOV-3 cells. Finally, we found that MY-03-01 likely induced SKOV-3 apoptosis by activating caspase3 and caspase9 through the mitochondrial pathway.

• 한국미생물·생명공학회지
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• 제23권3호
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• pp.255-262
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• 1995

Intact cells of Acinetobacter sp. T5-7 completely degraded trichloroethylene (TCE) following growth with phenol. This strain could grow on at least eleven aromatic compounds, e.g., benzaldehyde, benzene, benzoate, benzylalochol, catechol, caffeic acid, 2.4-D, p-hydroxybenzoate, phenol, protocatechuate and salicylate, and did grow on alkane, such as octane. But except phenol, other aromatic compounds did not induced TCE degradation. Phenol biotransformation products, catechol was identified in the culture media. However, catechol-induced cells did not degrade TCE. So we assumed that phenol hydroxylase was responsible for the degradation of TCE. The isolate T5-7 showed growth in MM2 medium containing sodium lactate and catechol rather than phenol, but did not display phenol hydroxyalse activity, suggesting induction of enzyme synthesis by phenol. Phenol hydroxylase activity was independent of added NADH and flavin adenine dinucleotide but was dependent on NADPH addition. Degradation of phenol produced catechols which are then cleaved by meta-fission. We identified catechol-2.3-dioxygenase by active staining of polyacrylamide gel.

• 한국미생물·생명공학회지
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• 제27권2호
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• pp.124-128
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• 1999

TK6 was able to produce NAD+ dependent cyclohexanol dehydrogenase(CDH). The production of CDH was increased rapidly at the logarithmic phase and maintained constantly after that. In order to investigate the inductive production of CDH by various substrates, the bacteria were grown in the media containing alicyclic hydrocarbons and various alcohols as a sole crabon souce. CDH was induced most actively by cyclohexanol. Cyclohexanone and cyclohexane-1,2-diol also induced remarkable amount of CDH but it was induced weakly by 1-propanol, 1-butanol, 1-pentanol, 1-hexanol, 2-propanol, and 2-methyl-1-propanol. The dehydrogenase of the bacteria grown in the media containing cyclohexanol were weakly active for various alcohols, but the dehydrogenase activity for cyclohexane-1,2-diol was twice as much as that for cyclohexanol. Activity staining on PAGE of the cell free extract of Rhodococcus sp. TK6 grown in the media containing cyclohexanol reveals at least sever isozyme bands of CDH and we nominated the four major activity bands as CDH I, II, III, and IV. CDH I was strongly induced by cyclohexanol, cyclohexane-1,2-diok, but its activity was specific to cyclohexane-1,2-diol and 1-pentanol. CDH IV was strongly induced by cyclohexanol and cyclohexane-1,2-diol, and its activity was very specific to cyclohexane-1,2-diol.

• The Korean Journal of Physiology and Pharmacology
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• 제1권4호
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• pp.355-366
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• 1997

Although cerebellar Purkinje cells display spontaneous electrical activity in vivo and in slice experiments, the mechanism of the spontaneous activity generation has not been clearly understood. The aim of this study was to investigate whether cerebellar Purkinje cells of postnatal rats generate spontaneous electrical activity without synaptic inputs. Dissociated cerebellar Purkinje cells were used for reducing synaptic inputs in the present study. Cerebellar Purkinje cells with dendrites were dissociated from postnatal rats using enzymatic treatment followed by mechanical trituration. Spontaneous electrical activities were recorded from dissociated cells without any stimulus using whole-cell patch clamp configuration. Two types, spontaneously firing or quiescent, of dissociated Purkinje cells were observed in postnatal rats. Both types of cells were identified as Purkinje cells using immunocytochemical staining technique with anti-calbindin after recording. Spontaneously active cells displayed two patterns of firing, repetitive and burst firings. Two thirds of dissociated Purkinje cells displayed repetitive firing and the rest of them did burst firing under same recording condition. Repetitive firing activities were maintained even after further isolation using either physical or pharmacological techniques. Neither high magnessium solution nor excitatory synaptic blockers, AP-5 and DNQX, block the spontaneous activity. These results demonstrate that spontaneous electrical activity of isolated cerebellar Purkinje cells in postnatal rats is generated by intrinsic membrane properties rather than synaptic inputs.

• Molecules and Cells
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• 제22권1호
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• pp.21-29
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• 2006

The aim of this study was to determine if hydrogen peroxide ($H_2O_2$) generated by glucose oxidase (GO) induces apoptosis or necrosis of BJAB cells and which radical is the direct mediator of cell death. We found that GO produced $H_2O_2$ continuously in low concentrations, similar to in vivo conditions, and decreased proliferation and cell viability in a dose-dependent manner. The GO-mediated cytotoxicity resulted from apoptosis, and was confirmed by monitoring the cells after H33342/Annexin V/propidium iodide staining. Decreases of mitochondrial membrane potential and intracellular glutathione level were found to be critical events in the $H_2O_2$-mediated apoptosis. Additional experiments revealed that $H_2O_2$ exerted its apoptotic action through the formation of hydroxyl radicals via the Fenton rather than the Haber-Weiss reaction. Moreover, intracellular redox-active iron, but not copper, participated in the $H_2O_2$-mediated apoptosis.