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Role of Amino Acid Residues within the Disulfide Loop of Thanatin, a Potent Antibiotic Peptide

  • Lee, Myung-Kyu;Cha, Li-Na;Lee, Si-Hyung;Hahm, Kyung-Soo
    • BMB Reports
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    • v.35 no.3
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    • pp.291-296
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    • 2002
  • Thanatin, a 21-residue peptide, is an inducible insect peptide with a broad range of activity against bacteria and fungi. It has a C-terminal disulfide loop, like the frog skin secretion antimicrobial peptides of the brevinin family. In this study, we tried to find the effect of a number of amino acids between the disulfide bond. Thanatin showed stronger antibacterial activity to Gram negative bacteria than other mutants, except Th1; whereas, the mutant peptides with deletion had higher activity to Gram positive bacteria than thanatin. An increase in the number of amino acid(s) using the alanine residue decreased the antibacterial activity in all of the bacteria. Th1 with deletion of threonine at position 15 ($Thr^{15}$) showed similar antibacterial activity against Gram-negative bacteria, but had higher activity against the Gram positive bacteria. In order to study the structure-function relationship, we measured liposome disruption by the peptides and CD spectra of the peptides. Th1 also showed the highest liposome leaking activity and α-helical propensity in the sodium dodecyl sulfate solution, compared with other peptides. Liposome disruption activity was closely correlated with the anti-Gram positive bacterial activity. All of the peptides showed no hemolytic activity. Th1 was considered to be useful as an antimicrobial peptide with broad spectrum without toxicity.

Beneficial Effect of DA-9601, an Extract of Artemisiae Herba, on Animals Models of Inflammatory Bowel Disease

  • Ahn, Byoung-Ok;Ryu, Byong-Kweon;Ko, Jun-Il;Oh, Tae-Young;Kim, Soon-Hoe;Kim, Won-Bae;Yang, Jun-Nick;Lee, Eun-Bang;Hahm, Ki-Baik
    • Biomolecules & Therapeutics
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    • v.5 no.2
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    • pp.165-173
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    • 1997
  • This study was conducted to investigate the effect of DA-9601, an extract of Artemisiae Herba, which is known to possess mucoprotective action either by free radical scavenging effect or increase of mucus secretion, against animal models of inflammatory bowel disease (IBD) induced by trinirobenzene sulfonic acid (TNBS) or other noxious agents. Experimental colitis was induced by intracolonic administration of TNBS in 50% ethanol, or 1 ml of 7% acetic acid solution (AA), by subcutaneous injection of indomethacin (INDO) in rats, or by supplementing drinking water with 5% dextran sodium sulfate (DSS) in albino mice. DA-9601 was treated orally for 4 to 7 days. Animals were euthanized 1 day after the last treatment for morphological and biochemical analysises. All the noxious agents including TNBS, AA, INDO and DSS elicited severe colitis. The animals treated with DA-9601 showed a consistent, dose-related reduction in the severity of colitis, grossly and histologically. The reduction was significant (p<0.05) after administration of DA-9601 at dose range of 10 mg/kg or above. In TNBS-induced colitis, the rats receiving DA-9601 showed significantly decreased mucosal myeloperoxidase (MPO) and thiobarbituric acid-reactive substances (TBA-RS), when compared to control and mesalazine groups. Mucosal proinflammatory cytokine levels were also decreased after DA-9601 treatment. In conclusion, DA-9601 ameliorated macroscopic and histologic scores in experimental colitis either through decreasing oxidative stress or by attenuating cytokines involved in inflammation. DA-9601 could be a promising drug for the therapy of IBD.

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The Change of Salivary and Oral Bacteria Amount by Composition of SLS Contents of Toothpaste (세치제내 SLS함유에 따른 타액과 세균변화)

  • Shim, Youn-Su;Jeong, Mi-Ae;Jeong, Sang-Hee
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.9
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    • pp.3341-3346
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    • 2010
  • Dentifrices comtaining SLS of 2.2% and 0.0% were applied to 60 subjects for 4 weeks in this study. Dentifrice is used for salivary and oral bacteria amount change according to its SLS contents in toothpaste. The results were as follows. Anaysis of LSL contents befor and after the after experiment in salivary and bacterial count oral cavity didn't show significant difference in time interval. There was no correlation between salivary flow rate and bacterial count in the oral cavity according to content of SLS. Dentifrice of 2.2% content satisfied the subjects much more than that of 0.0% content(p<0.001).

Simplified HPLC Method for the Determination of Pseudoephedrine Hydrochloride from Allegra D Tablet

  • Park, Moon-Hee;Shin, In-Chul
    • Biomolecules & Therapeutics
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    • v.15 no.2
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    • pp.123-126
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    • 2007
  • A sensitive, simple and highly selective liquid chromatography method of determination for extraction of pseudoephedrine hydrochloride from Allegra D tablet was developed. The chief benefit of the present method is the minimal sample preparation, as the procedure is only filtering through pore syringe filter. Two drugs (pseudoephedrine hydrochloride, fexofenadine) were separated on a C$_{18}$ column and analyzed by high performance liquid chromatography (HPLC). The method had a chromatographic run time of 8.0 min. 1 ml of pseudoephedrine hydrochloride solution (1 mg/ml) was filtered through 0.22 um pore syringe filter. 50 ul of filtering solution was injected to HPLC pump and we knew the retention time (1.85 min) of separating of pseudoephedrine hydrochloride using UV detector at 280 nm. We used C$_{18}$ column (4.6 mm${\times}$250 mm), mobile phase solution (<0.05 mol/L NaH$_2$PO$_4$, 2 ml/L H$_3$PO$_4$>/CH$_3$CN / sodium dodesyl sulfate = 60 ml / 40 ml / 1 g). We separated psedoephedrine hydrochloride at run time of 1.85 min from Allegra D tablet solution (1 mg/ml) filtered through 0.22 um pore syringe filter using UV detector at 280 nm. Flow rate was set at 1.0 ml/min and the column temperature was set at 40$^{\circ}C$. Psedoephedrine hydrochloride solution (1 mg/ml) separated from Allegra D tablet was filtered through 0.22 um pore syringe filter and injected 50 ul. We confirmed the peak of psedoephedrine hydrochloride at same retention time and the separating solution was freeze-dried. In conclusion, A simple isocratic reverse-phase HPLC method has been developed that provides excellent separation of pseudoephedrine from Allegra D tablet.

Production of Xylooligosaccharides with Thermostable Xylanases from the Streptomyces thermocyaneo-violaceus (내열성 방성균 Streptomyces thermocyaneoviloaceus 의 Xylanases를 이용한 자일로올리고당의 생산)

  • 이오석;최충식;최준호;주길재;이인구
    • Microbiology and Biotechnology Letters
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    • v.29 no.4
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    • pp.221-226
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    • 2001
  • Streptomyces themocyaneovio-laceus producing the thermostable xylanase was used for the production of xylooligosaccharides from xylan. The optimal conditions for the xylanase production were investigated in jar fermentor, which operated at 2 vvm aera-tion and 400 rpm agitation speed at $50^{\circ}C$ for 24 h. The optimal reaction condtion for the production of xylooli-gosaccharides with xylanases which were prepared by the percipitation with ammonium sulfate were obtained by the reaction at $60^{\circ}C$ for 12 h in the mixture composed of 10% birchwood xylan in 50 mM sodium phosphate buffer (pH 6.0)and 10 unit/ml of xylanase. In this optimal condition for the xylooligosaccharides production the mixture of xylooligosaccharides (58.8 g/I) which were composed of 20.1 g/I of xyobiose, 8.9 g/I of xylotriose 4.5 g/I of xylotetraose 16.2g/I of xylopentaose and 9.1 g/I xylohexaose and 5.0 g/I of xylose was produced from 100 g/I of birchwood xylan by the xylanases of S thermocyaneoviolaceus .

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Studies on the Cellulolytic Enzyme System of Rhizopus sp. G-211 Isolated from Rotting Ginseng (인삼부패란중 Rhizopus sp. G-211이 생성하는 Cellulase 에 관한 연구)

  • 노혜원;김상달;도재호;강성호
    • Microbiology and Biotechnology Letters
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    • v.10 no.1
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    • pp.1-7
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    • 1982
  • A Rhizopus sp. was selected for its strong cellulolytic activity among various strains of molds found in rotting ginseng roots. Studies were made on some properties of the cellyloiytic enzyme produced by the strain. The results obtained were summarized as follows: The optimum pH of the enzyme was 4.5 and the range of its stability to the pH was 3.0 to 7.0. The optimum temperature was 5$0^{\circ}C$, while the enzyme was instantly inactivated above 6$0^{\circ}C$. Mn$^{++}$ and Co$^{++}$ ions increased enzyme activity and the metal ions were found to increased the ther-mostability of the enzyme. This enzyme was inhibited by sodium dodecyl sulfate and 2,4-dinitrophenol. This enzyme had a strong cellulolytic enzyme activity on various native cellulose given a sufficient reaction time. The addition of 0.5% saponin solution into reaction mixture increased the enzyme activity.

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Evaluation of Local Lymph Node Assay as an Alternative Method for Skin Sensitization Potential in Baltic Mice (Balb/c 마우스에서 Local Lymph Node Assay(LLNA)를 01용한 피부 감작성 시험 대체시험법 연구)

  • 이종권;황인창;박재현;김형수;정승태;엄준호;오혜영
    • Toxicological Research
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    • v.18 no.2
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    • pp.175-181
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    • 2002
  • Allergic contact dermatitis (skin sensitization) may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. This study was carried out to evaluate the skin sensitization potential for chemicals in Balb/c mice by LLNA. Contact allergen, dinitrochlorobenzene (DNCB), respiratory allergen, toluene diisocyanate (TDI) and a weak allergen, $\alpha$-hexlycinnamaldehyde (HCA) were wed as positive chemicals and irritant, sodium lauryl sulfate(SLS) also wed as a reference chemical in this study. The weights of lymph node in the mice treated with DNCB, TDI, and HCA were increased compared to vehicle control. There was a significant increase in lymph node weight of mice treated with high concentration of SLS compared to vehicle control. The stimulation index (SI) of Lymph node cell in the mice treated with DNCB, TDI, and HCA revealed over three-fold increase compared to vehicle control by $3^H$-thymidine uptake. All allergens correctly identified in this LLNA study wing Balb/c mice. These results suggest that LLNA wing Balb/c mice could be a useful method for screening the allergenic potential of chemicals. The expression of IL-2 mRNA was slightly increased in draining auricular lymph node cell of the mice treated with TDI and HCA by RT-PCR. However the IL-2 levels in DNCB and SLS of treated animals were not significantly changed.

Electrokinetic Property and Flotation Characteristics of Scheelite (灰重石의 水溶液中에 있어서의 界面現象과 浮選特性에 關한 硏究)

  • Hyung Sup Choi;Kook Nam Han
    • Journal of the Korean Chemical Society
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    • v.7 no.1
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    • pp.17-24
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    • 1963
  • The fundamental investigations of surface properties of scheelite were made by electrophoretic mobility adsorption and contact angle measurements, and results have been correlated with its floatability obtained by Hallimond tube flotation test. The role of the interfacial electrical condition on the adsorption of collectors on mineral surfaces is discussed with the flotation of scheelite. From electrokinetic measurements made on scheelite, $Ca^{++}$ and $WO_4^{--}$ are identified to act as potential-determining ions, thus controlling the surface properties on this mineral. Therefore, at the fixed pH, the scheelite surface become to be less negatively charged with increasing $Ca^{++}$ concentration and more negatively charged with increasing $WO_4^{--}$ concentration in the pulp. Adsorption of collectors then depends strongly on the concentration of $Ca^{++}$ or $WO_4^{--}$ in the solution; anionic collectors are adsorbed on less negatively charged surfaces and cationic collectors on more negatively charged surfaces, which in turn defines the effective flotation range with respective collectors for this mineral.

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Oral administration of fermented wild ginseng ameliorates DSS-induced acute colitis by inhibiting NF-κB signaling and protects intestinal epithelial barrier

  • Seong, Myeong A;Woo, Jong Kyu;Kang, Ju-Hee;Jang, Yeong Su;Choi, Seungho;Jang, Young Saeng;Lee, Taek Hwan;Jung, Kyung Hoon;Kang, Dong Kyu;Hurh, Byung Seok;Kim, Dae Eung;Kim, Sun Yeou;Oh, Seung Hyun
    • BMB Reports
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    • v.48 no.7
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    • pp.419-425
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    • 2015
  • Ginseng has been widely used for therapeutic and preventive purposes for thousands of years. However, orally administered ginseng has very low bioavailability and absorption in the intestine. Therefore, fermented ginseng was developed to enhance the beneficial effects of ginseng in the intestine. In this study, we investigated the molecular mechanisms underlying the anti-inflammatory activity of fermented wild ginseng (FWG). We found that FWG significantly alleviated the severity of colitis in a dextran sodium sulfate (DSS)-induced colitis mouse model, and decreased expression level of pro-inflammatory cytokines in colonic tissue. Moreover, we observed that FWG suppressed the infiltration of macrophages in DSS-induced colitis. FWG also attenuated the transcriptional activity of nuclear factor-κB (NF-κB) by reducing the translocation of NF-κB into the nucleus. Our data indicate that FWG contains anti-inflammatory activity via NF-κB inactivation and could be useful for treating colitis. [BMB Reports 2015; 48(7): 419-425]

Effect of Melatonin on Differentiating 3T3-L1 Preadipocytes (3T3-L1 지방전구세포의 지방분화에서 멜라토닌의 영향)

  • Lee, Jeongkun;Lee, Yeong Hun;Kim, Chi Hyun
    • Journal of Biomedical Engineering Research
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    • v.41 no.3
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    • pp.138-145
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    • 2020
  • Adipocytes are the main constituent of adipose tissue. Understanding the molecular basis of adipogenesis is pivotal to finding the therapeutic targets for treatment of obesity. Melatonin is associated with obesity and its mechanism is currently under intensive investigation. The objective of this study was to investigate the effect of melatonin on adipogenesis in differentiating preadipocytes. 3T3-L1 preadipocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) containing 5% calf serum at 37℃ with 5% CO2 in a humidified incubator. Differentiation was induced using DMEM with 10% fetal bovine serum supplemented with MDI two days after cell confluence (day 0). Cells were treated with 0, 10 and 100 μM melatonin on either day 0 or day 5. 72 hours after each treatment, lipid accumulation was measured by oil red O staining. Proteins were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to membranes. As a result, lipid accumulation decreased with melatonin treatment. ERK pathway, activated when differentiation is induced, also decreased with an increase in melatonin concentration. Furthermore, the expression of key adipogenic factors, C/EBPα, C/EBPβ, and PPARγ, were reduced by melatonin treatment. These results imply that melatonin may inhibit the process of adipogenesis and may have a role as a new anti-obesity agent.