• Korean Journal of Microbiology
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• v.45 no.4
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• pp.362-367
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• 2009

Among total 176 strains with antialgal effects isolated from So-ok stream in Korea, Pseudomonas aeruginosa AJ1 showed the highest removal efficiency for an algal species Microcystis aeruginosa (clear zone of diameter 50.0 mm on algal lawn after 20 days). The algal growth was suppressed even when the supernatant of AJ1 culture was applied, suggesting that extracellular substances are responsible for its antialgal activity. The removal activity of AJ1 was optimal under the following condition: pH 8, $30^{\circ}C$, and mannitol as a carbon source. The antialgal activity of AJ1 appeared to be dependent of the growth phase of M. aeruginosa, i.e., the highest at the early phase, but not its own phase. As expected, the algicidal effect was improved as the amount of the treated supernatant was increased; the highest removal efficiency (80.3%) was achieved when 40 ml/L of the supernatant was used. Interestingly, however, the removal rate was opposite. The highest removal rate ($8.2{\mu}g$ chl-a/ml supernatant/day) was achieved when low concentration (10 ml/L) was applied. These results suggest that P. aeruginosa AJ1 is a promising biological agent to control the problematic algal bloom.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.277-285
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• 2001

Periodontal disease is characterized by inflammation and subsequent loss and/or damage to tooth-supporting tissues such as bone, cementum,and periodontal ligament. Periodontal ligament and cementum are the key tissues in the initial process of regeneration following periodontal disease. Therefore, studies on cementoblasts, which form cementum are emphasized. It is still unclear which cells cementoblast differentiate from. This study was conducted under the hypothesis that PDL fibroblast can differentiate into either cementoblast or osteoblast depending on the conditions of surrounding tissue. Clinically, with excessive traction force of orthodontic appliances or excessive occlusion hypercementosis is observed, and this has been confirmed histologically. Consequently, activation of cementoblast can be expected in rats when mechanical stimuli are given to PDL fibroblast. Therefore, the purpose of this article is to prove that PDL fibroblast differentiates into cementoblast in rats under mechanical stimuli using histologic and molecular methods. In this study, twenty rats were given hard diet. Ten of them were sacrificed after 1 week, and the others were sacrificed after two weeks. Slides were made from tooth specimen, and they were studied under the microscope. In addition, PDL fibroblast and cementum from the extracted teeth were analyzed with Northern blotting. In histologic examination, as time passed, PDL fibroblast migrated to the dentin side, differentiated into cementoblast, and formed new cementum. In Northern blotting, it was found that mRNA expression of cementoblast-specific proteins such as BSP, OC, OPN, and type I collagen were more prominent in rats sacrificed after 2 weeks of hard-diet than rats sacrificed after 1 week. From these findings we can conclude that PDL fibroblast can differentiate into cementoblast under mechanical stimuli. We think that 'Rat Models' used in this study will be beneficial to future studies regarding cementoblast.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1628-1638
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• 2017

Viola tianshanica Maxim, belonging to the Violaceae plant family, is traditionally used in Uighur medicine for treating pneumonia, headache, and fever. There is, however, a lack of basic understanding of its pharmacological activities. This study was designed to observe the effects of the ethanol extract (TSM) from Viola tianshanica Maxim on the inflammation response in acute lung injury (ALI) induced by LPS and the possible underlying mechanisms. We found that TSM (200 and 500 mg/kg) significantly decreased inflammatory cytokine production and the number of inflammatory cells, including macrophages and neutrophils, in bronchoalveolar lavage fluid. TSM also markedly inhibited the lung wet-to-dry ratio and alleviated pathological changes in lung tissues. In vitro, after TSM ($12.5-100{\mu}g/ml$) treatment to RAW 264.7 cells for 1 h, LPS ($1{\mu}g/ml$) was added and the cells were further incubated for 24 h. TSM dose-dependently inhibited the levels of proinflammatory cytokines, such as NO, $PGE_2$, $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$, and remarkably decreased the protein and mRNA expression of $TNF-{\alpha}$ and IL-6 in LPS-stimulated RAW 264.7 cells. TSM also suppressed protein expression of $p-I{\kappa}Ba$ and p-ERK1/2 and blocked nuclear translocation of $NF-{\kappa}B$ p65. The results indicate that TSM exerts anti-inflammatory effects related with inhibition on $NF-{\kappa}B$ and MAPK (p-ERK1/2) signaling pathways. In conclusion, our data demonstrate that TSM might be a potential agent for the treatment of ALI.

• Polymer(Korea)
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• v.33 no.1
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• pp.26-32
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• 2009

The aim of this research was to prepare microparticulate systems based on poly (lactide-co-glycolide)(PLGA) for the local release of ipriflavone in order to reduce bone loss. We developed the IP loaded PLGA microspheres using relatively simple oil-in-water(O/W) solvent evaporation method. HPLC was used to perform the in vitro release test of IP and morphology of cell attached on the micro-spheres was investigated using SEM. Cytotoxicity was assayed by cell counting kit-8 (CCK-8) test. Osteogenic differential cells were analyzed by ALP activity. Through RT-PCR analysis, we observed osteocalcin, ALP, and Type I collagen mRNA expression. The release of IP in vitro was more prolonged over 42 days and IP/PLGA microspheres showed the improvement on the cell proliferation, ALP activity and RT-PCR comparing with control (only PLGA). This initial research will be used to direct future work involved in developing this composite injectable bone tissue engineering system.

• Journal of Sericultural and Entomological Science
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• v.49 no.2
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• pp.37-42
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• 2007

Lactoferrin, an ion-binding 80-kDa glycoprotein, has been suggested to have many biologic activities, such as facilitating ion absorption and having antimicrobial and anti-inflammatory effects. Several of these activities are likely to only be facilitated by human lactoferrin because they depend on the binding of human lactoferrin to specific receptor. To produce recombinant human lactoferrin to animal foods using transgenic silkworm, Bombyx mori L, we have cloned and sequenced the cDNA encoding for a human lactoferrin (HLf) from the mRNA in mammary tumor line (GI-101). As a result, the 2.5-kb fragment of HLf gene was cloned with pGEM-T vector and then this fragment was sequenced. In the nucleotide sequence analysis, single open reading frame of the 2,136-bp encoding for a polypeptide of 712 amino acid residues was detected. On the other hand, we constructed a recombinant plasmid(pPT-HLf), containing human lactoferrin gene for germ line transformation of the silkworm using a piggyBac transposon-derived vector. A nonautonomous helper plasmid encodes the piggyBac transposase. Approximately 6.7% of individuals in the G0 silkworms expressed green fluorescent protein (GFP). PCR analyses of GFP-positive silkworms (G0 and G1) revealed that independent insertions occurred frequently. Furthermore, Western blot analysis showed that the recombinant HLf expressed in hemolymph has the same molecular weight (80 kDa) as a native protein. On the basis of these experiments, expression of HLf in next generation of transgenic silkworm is now in process.

• Korean Journal of Veterinary Research
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• v.35 no.3
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• pp.515-530
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• 1995

Coronaviridae에 속하는 transmissible gastroenteritis virus(TGEV)와 porcine epidemic diarrhea virus(PEDV)를 specific하게 detection할 수 있는 방법을 개발하고자 본 연구를 수행하였다. 두 바이러스 모두 RNA 바이러스이기 때문에 reverse transcription-polymerase chain reaction(RT-PCR)으로 nucleocapsid(N) protein gene의 cDNA를 증폭시켰다. SmaI으로 처리한 pTZ19R에 ligation시킨 후 염기서열을 밝히고자 sequencing하였다. 각각의 prototype virus와 비교하여 상동성을 밝혔다. 두 바이러스에 대한 cDNA probe를 제작하여 Southern blot hybridization을 실시하였다. TGEV의 경우 백신주인 P45와 병독주인 Miller strain을 사용하였다. cDNA를 증폭시키기 위해 N1/N1R과 N2/N2R 두 가지 primer를 이용한 결과, N1/N1R primer의 경우 586bp 크기의 PCR product를 얻을 수 있었고, N2/N2R primers로 582bp의 cDNA를 증폭시킬 수 있었다. PEDV 실험을 위하여 PED 임상 증상을 나타내는 분변을 이용하여 RT-PCR을 실시하였다. P2/P2R primer로 753bp의 PCR product를 얻을 수 있었다. TGEV의 두 가지 strain의 N protein gene을 sequencing하여 prototype인 Purdue strain과 염기서열 상동성을 조사한 결과, 97%이상의 높은 homology를 나타내었다. PED-V 역시 N protein gene을 sequencing하여 CV777과 염기서열 상동성을 조사한 결과 97%이상의 homology로 PEDV임을 알 수 있었다. TGEV와 PEDV의 염기서열을 비교한 결과 29%의 낮은 homology를 관찰할 수 있었다. 두 가지 바이러스의 N protein gene에 대한 cDNA probe를 제작하여 Southern blot hybridization을 한 결과, 각 바이러스에 매우 특이적 반응을 나타내었다.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.38-45
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• 2018

Acne is an inflammatory skin disease that occurs in puberty and young people. Propionibacterium acnes (P. acnes) is known to be a major cause of inflammation in acne. P. acnes proliferates within hair follicles blocked by overproduced sebum in the skin, and thereby activates monocytic cells to promote the secretion of pro-inflammatory cytokines. In this study, we investigated the possibility of Cnestis palala (Lour.) Merr. extract to diminish P. acnes-mediated inflammatory responses. We found that C. palala extract significantly attenuated P. acnes-induced pro-inflammatory cytokine expressions, such as $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, iNOS, and COX-2 in mouse macrophage RAW264.7 cells. Moreover, we observed that C. palala extract inhibited $NF-{\kappa}B$ transcriptional activation, which is the major transcription factor of inflammatory cytokine expression. Therefore, it is expected that C. palala extract has a potential as a therapeutic agent or supplement for the treatment P. acnes-induced inflammatory responses.

• The Korean Journal of Mycology
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• v.26 no.3 s.86
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• pp.354-360
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• 1998

We isolated three amplified DNA fragments from P. sajor-caju by Polmerase chain reaction (PCR) using the chithin synthase specific primers. Since the sequence analysis of the these fragments showed significant homology to the other known chitin synthase gene, we regarded these cloned fragments as PsCHS1, PsCHS2, and PsCHS3 according to their size. The PsCHS3, which showed the highest sequence homology (83% identity in amino acid level with ChsI of Rhizopus oligosporus in conserved region), was selected to see expression pattern of the corresponding gene. The result of RT-PCR using internal primer of the PsCHS3 fragment revealed that PsCHS3 gene was only expressed in cap and mycelium but not in stipe. In order to see whether the PsCHS3 gene was to be induced by wounding, the comparison of the mRNA level of this gene between wounded and unwounded mature cap showed at least two times induction of this gene by wounding treatment.

• Molecules and Cells
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• v.27 no.4
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• pp.423-428
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• 2009

Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.87-94
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• 2017

Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.