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Development and Characterization of an Anti-Acne Gel Containing Siamese Crocodile (Crocodylus siamensis) Leukocyte Extract

  • Phupiewkham, Weeraya;Lu, Qiumin;Payoungkiattikun, Wisarut;Temsiripong, Threeranan;Jangpromma, Nisachon;Lai, Ren;Klaynongsruang, Sompong
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.707-717
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    • 2018
  • Leukocytes are reportedly the first line of the innate immune defense and essential for the control of common bacterial infections. Therefore, in this work, the antibacterial activity of crocodile leukocyte extract against Propionibacterium acnes was evaluated, and we also characterized the related activity of skin infection. The leukocyte extract showed the minimum inhibitory concentration to be $100{\mu}g/ml$ to P. acnes. SEM imaging demonstrated that the leukocyte extract adversely affected P. acnes cell permeability in a concentration-dependent manner. Furthermore, the crocodile leukocyte extract could significantly reduce proinflammatory markers and decrease inflammatory signs in infected mouse ears. The crude leukocyte extract was further purified using FPLC and RP-HPLC. The resulting fraction F5 was indicated as the anti-acne peptide-containing fraction. The molecular mass of the peptide contained in F5 was calculated to be 4,790.5 Da. N-Terminal sequencing revealed the amino acid sequence as GPEPVPAIYQ, which displays similarities to immunoglobulin A and leucine-rich repeat neuronal protein. This is the first reported amino acid sequence of a crocodile leukocyte extract that possesses anti-acne activity. To attempt to use it in a prototype cosmetic, an anti-acne gel containing crude crocodile leukocyte extract was formulated, resulting in seven gel formulations (G1, G2, G3, G4, G5, G6, and G7). The formulations G5, G6, and G7 exhibited 2-fold higher anti-acne activity than G1-G4. Investigation of accelerating stability studies of anti-acne gel formulations G5, G6, and G7 demonstrated that a low storage temperature ($4^{\circ}C$) is suitable for maintaining the physical properties and biological activity of the anti-acne gel products.

Characteristics of $Ta_{2}O_{5}$ Films by RF Reactive Sputtering (RF 반응성 스펏터링으로 제조한 $Ta_{2}O_{5}$ 막의 특성)

  • Park, Wug-Dong;Keum, Dong-Yeal;Kim, Ki-Wan;Choi, Kyu-Man
    • Journal of Sensor Science and Technology
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    • v.1 no.2
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    • pp.173-181
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    • 1992
  • Tantalum pentoxide($Ta_{2}O_{5}$) thin films on p-type (100) silicon wafer were fabricated by RF reactive sputtering. Physical properties and structure of the specimens were examined by XRD and AES. From the C-V analysis, the dielectric constant of $Ta_{2}O_{5}$ films was in the range of 10-12 in the reactive gas atmosphere in which 10% of oxygen gas is mixed. The ratio of Ta : 0 was 1 : 2 and 1 : 2.49 by AES and RBS examination, respectively. The heat-treatment at $700^{\circ}C$ in $O_{2}$ ambient led to induce crystallization. When the heat-treatment temperature was $1000^{\circ}C$, the dielectric constant was 20.5 in $O_{2}$ ambient and 23 in $N_{2}$ ambient, respectively. The crystal structure of $Ta_{2}O_{5}$ film was pseudo hexagonal of ${\delta}-Ta_{2}O_{5}$. The flat band voltage shift(${\Delta}V_{FB}$) of the specimens and the leakage current density were decreased for higher oxygen mixing ratio. The maximum breakdown field was 2.4MV/cm at the oxygen mixing ratio of 10%. The $Ta_{2}O_{5}$ films will be applicable to hydrogen ion sensitive film and gate oxide material for memory device.

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Cordycepin-Enriched WIB801C from Cordyceps militaris Inhibits Collagen-Induced [Ca2+]i Mobilization via cAMP-Dependent Phosphorylation of Inositol 1, 4, 5-Trisphosphate Receptor in Human Platelets

  • Lee, Dong-Ha;Kim, Hyun-Hong;Cho, Hyun-Jeong;Yu, Young-Bin;Kang, Hyo-Chan;Kim, Jong-Lae;Lee, Jong-Jin;Park, Hwa-Jin
    • Biomolecules & Therapeutics
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    • v.22 no.3
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    • pp.223-231
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    • 2014
  • In this study, we prepared cordycepin-enriched (CE)-WIB801C, a n-butanol extract of Cordyceps militaris-hypha, and investigated the effect of CE-WIB801C on collagen-induced human platelet aggregation. CE-WIB801C dose-dependently inhibited collagen-induced platelet aggregation, and its $IC_{50}$ value was $175{\mu}g/ml$. CE-WIB801C increased cAMP level more than cGMP level, but inhibited collagen-elevated $[CA^{2+}]_i$ mobilization and thromboxane $A_2$ ($TXA_2$) production. cAMP-dependent protein kinase (A-kinase) inhibitor Rp-8-Br-cAMPS increased the CE-WIB801C-downregulated $[CA^{2+}]_i$ level in a dose dependent manner, and strongly inhibited CE-WIB801C-induced inositol 1, 4, 5-trisphosphate receptor ($IP_3R$) phosphorylation. These results suggest that the inhibition of $[CA^{2+}]_i$ mobilization by CE-WIB801C is resulted from the cAMP/A-kinase-dependent phosphorylation of $IP_3R$. CE-WIB801C suppressed $TXA_2$ production, but did not inhibit the activities of cyclooxygenase-1 (COX-1) and $TXA_2$ synthase (TXAS). These results suggest that the inhibition of $TXA_2$ production by WIB801C is not resulted from the direct inhibition of COX-1 and TXAS. In this study, we demonstrate that CE-WIB801C with cAMP-dependent $CA^{2+}$-antagonistic antiplatelet effects may have preventive or therapeutic potential for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.

Korean Red Ginseng-intake has Definite Clinical Usefulness and causes Nef Gene Variation including High Frequency of Deletion

  • Cho Young Keol;Lee Hee Kyung;Ahn Sun Hee;Lee Hee Jung;Nam Ki Yeul
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.185-211
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    • 2002
  • We have found many beneficial effects of the long-tenn intake of Korean red ginseng (KRG) in human immunodeficiency virus (HIV) type-I infected patients, including the maintenance of CD4+ T cell count for 10 years with KRG only and the delayed development of resistance mutation to ZDV. In this study, to investigate whether KRG-intake could affect the clinical progression and nef gene variation, we determined 200nef sequences from 70 patients. Follow-up period was $8.8{\pm}2.9$ years and annual decrease in CD4+T cell was $41{\pm}57/ul.$ Nested polymerase chain reaction (PCR) and direct sequencing were perfonned with peripheral blood mononuclear cells (PBMC) obtained at times during the study period. First, there was a significant correlation between survival duration and duration of KRG-intake $(36.8{\pm}38$ months)(P=0.000). There were significant correlations between the last NefProg score and CD4+ T cell count (r= 0.208, P<0.05) and annual decrease in CD4+ T cell count (r =0.346, P<0.01) in 70 patients. In addition, there were significant correlations between KRG-intake and annual decrease (r= 0.323, P<0.01), and the CD4+ T cell count itself (r=0.229, p<0.05). Furthennore, there was also a mild significance between the NefProg score and the duration of KRG-intake in only SP and RP (n=30, r=-0.281, P=0.067). In addition, we detected various defects in 21 patients $(30.0\%),$ not including 5 premature stop codons. Ten $(12.5\%)$ patients showed repeated deletion of an amino acid. Four of 10 patients were gross deletions and they were treated with KRG for more than 20 months. The number of patients with repeated gross deletions was significantly higher in the order of slow progressors $(18\%)$, typical progressors($3\%$), and rapid progressors($0\%$) (P<0.05). We also observed that long-tenn intake of KRG might make the change from A or D to T at position 54 and decrease NefProg score. Taken together, our results show clear evidence that the long-term intake of KRG has effects on nef gene variation as well as definite clinical usefulness.

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A Simple, Sensitive, and Specific HPLC Analysis of Tissue Polyamines using FNBT Derivatization: Its Application on the Study of Polyamine Metabolism in Regenerating Rat Liver (생체의 Polyamine-분석을 위하여 FNBT-유도체를 이용하는 간편하고 특이적이며 예민한 Isocratic RP-HPLC 분석법과 재생성 흰쥐-간의 Polyamine-대사의 변동에 관한연구)

  • Choi, Sang-Hyun;Kim, Hyung-Gun;Park, Hong-Ik;Chun, Boe-Gwun
    • The Korean Journal of Pharmacology
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    • v.24 no.2
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    • pp.233-240
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    • 1988
  • A simple and selective isocratic HPLC method for the analysis of tissue polyamine contents is described and applied on the study of the changes of the hepatic polyamine contents after partial hepatectomy in male rats. The hepatic polyamines are extracted with 0.4 M perchloric acid containing 2 mM disodium EDTA, and then the extract is redissolved in 100 ul of 1 M sodium carbonate and incubated with 300 ul of FNBT-dimethylsulfoxide (1: 100) mixture. The N-2'-nitro-4'-trifluoromethylphenyl drivatives of polyamines are separated through a ERC-ODS column in an isocratic mode with an acetonitrile-water (80:20) mobile phase within 20 min. per a sample, while monitoring the effluent at 242 nm. This improved method which could detect subnanogram of each polyamines is highly specific and reproducible as evidenced by the application of it on the study of the changes of polyamine contents in the regenerating rat liver after partial hepatectomy.

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Quantitative Analysis for Components of Epimedium koreanum (음양곽 주성분의 정량분석)

  • Han, Yong-Nam;Hwang, Keum-Hee;Lee, Mie-Soon
    • Korean Journal of Food Science and Technology
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    • v.28 no.4
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    • pp.616-623
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    • 1996
  • Eum Yang Kwak, the aerial part of Epimedium koreanum, is widely used as a folk medicine for stimulant in man, tonic, and hypotensive purpose. The plant contains icariin (a specific flavonoid), magnoflorine (an alkaloid) and tannin, but their contents are not known until now. In this paper, a quantitative analysis method for them was developed. Determination of icariin and magnoflorine was successfully achived by high performance liquid chromatography equipped with a UV detector in the ranges of $0.1{\sim}0.4\;mg$ and $0.002{\sim}0.1\;mg\;per\;ml$ sample, respectively. Extraction of the plant was carried out with water or 50% ethanol using different decocting temperatures and times. Icariin was well extracted either by water ($100^{\circ}C$, 3hr) or 50% ethanol ($85^{\circ}C$, 1hr), and its content in the plant was measured to be 0.94%. On the other hand, magnoflorine was fully extracted by 50% ethanol ($85^{\circ}C$, 1hr), and its content was determined to be 0.16%. Therefore, decoction of the medicinal plant with water at $100^{\circ}C$ for 3hr turned out to be recommendable for the best extraction.

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Nutrient Utilization, Body Composition and Lactation Performance of First Lactation Bali Cows (Bos sondaicus) on Grass-Legume Based Diets

  • Sukarini, I.A.M.;Sastradipradja, D.;Sutardi, T.;Mahardika, IG.;Budiarta, IG.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1681-1690
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    • 2000
  • A study on energy and protein utilization, and milk production of Bali cows on grass-legume diets was carried out using 12 first lactation cows (initial BW $263.79{\pm}21.66kg$) during a period of 16 weeks starting immediately post calving. The animals were randomly allotted into 4 dietary treatment groups R1, R2, R3 and R4, receiving from the last 2 months of pregnancy onwards, graded improved rations based on a mixture of locally available grass and legume feed ad libitum. R1 contained on a DM basis 70% elephant grass (PP, Penisetum purpureum) plus 30% Gliricidia sepia leaves (GS), R2 was 30% PP plus 55% GS supplemented with 15% Hibiscus tilliactus leaves (HT, defaunating effect), R3 and R4 were 22.5% PP+41.25% GS+11.25% HT+25% concentrate, where R3 was not and R4 supplemented with zinc di-acetate. TDN, CP and zinc contents of the diets were 58.2%, 12.05% and 18.3 mg/kg respectively for R1, 65.05%, 16.9% and 25.6 mg/kg respectively for R2, 66.03%, 16.71% and 29.02 mg/kg respectively for R3 and 66.03%, 16.71% and 60.47 mg/kg respectively for R4. Milk production and body weight were monitored throughout the experimental period. In vivo body composition by the urea space technique validated by the body density method and supported by carcass data was estimated at the start and termination of the experiment. Nutrient balance and rumen performance characteristics were measured during a balance trial of 7 days during the 3rd and 4th week of the lactation period. Results indicated that quality of ration caused improvement of ruminal total VFA concentration, increments being 52 to 65% for R2, R3 and R4 above R1, with increments of acetate being less (31 to 48%) and propionate being proportionally more in comparison to total VFA increments. Similarly, ammonia concentrations increased to 5.24 to 7.07 mM, equivalent to 7.34 to 9.90 mg $NH_3-N/100ml$ rumen fluid. Results also indicated that feed quality did not affect DE and ME intakes, and heat production (HP), but increased GE, UE, energy in milk and total retained energy (RE total) in body tissues and milk. Intake-, digestible- and catabolized-protein, and retained-protein in body tissues and milk (Rprot) were all elevated increasing the quality of ration. Similar results were obtained for milk yield and components with mean values reaching 2.085 kg/d (R4) versus 0.92 kg/d (R1) for milk yield, and 170.22 g/d (R4) vs 71.69 g/d (R1), 105.74 g/d (R4) vs 45.35 g/d (R1), 101.34 g/d (R4) vs 46.36 g/d (R1) for milk-fat, -protein, and -lactose, respectively. Relatively high yields of milk production was maintained longer for R4 as compared to the other treatment groups. There were no significant effects on body mass and components due to lactation. From the relationship $RE_{total}$ (MJ/d)=12.79-0.373 ME (MJ/d); (r=0.73), it was found that $ME_{m}=0.53MJ/kgW^{0.75}.d$. Requirement of energy to support the production of milk, ranging from 0.5 to 3.0 kg/d, follows the equation: Milk Prod. ($Q_{mp}$, kg/d)=[-2.48+4.31 ME($MJ/kg^{0.75}.d$)]; (r=0.6) or $Q_{mp}$=-3.4+[0.08($ME-RE_{body\;tissue}$)]MJ/d]; (r=0.94). The requirement for protein intake for maintenance ($IP_m$) equals $6.19 g/kg^{0.75}.d$ derived from the relationship RP=-47.4+0.12 IP; (r=0.74, n=9). Equation for protein requirement for lactation is $Q_{nl}$=[($Q_{mp}$)(% protein in milk)($I_{mp}$)]/100, where $Q_{nl}$ is g protein required for lactation, $Q_{mp}$ is daily milk yield, Bali cow's milk-protein content av. 5.04%, and $I_{mp}$ is metabolic increment for milk production ($ME_{lakt}/ME_{m}=1.46$).

Similarities of Scaritoxin to Ciguatoxin on the Chromatographic Behaviours (Scaritoxin과 Ciguatoxin의 크로마토그라피상에서의 몇가지 유사성)

  • Joh, Yong-Goe;Scheuer, Paul J.
    • Korean Journal of Food Science and Technology
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    • v.17 no.2
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    • pp.121-127
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    • 1985
  • In studying the structural work on ciguatoxin, parrot fish collected were identified as Scarus sordidus, S. frenatus, S. scaber and S. pectarlis, in which only S. sordidus contained toxic materials. Crude toxins obtained by silicic acid column chromatography, could be separated on a DEAE-cellulose column into two fractions, ST-1(less polar) and ST-2(polar) eluted with chloroform and chloroform-methanol(1:1). Furthermore ST-1 could be changed into ST-2 by repeated chromatography on DEAE-cellulose. Rf values of ST-1 and ST-2 were 0.60-0.75 and 0.30-0.54 on TLC coated with silica gel 60F-254 developed by chloroform-methanol-water-acetic acid (90:9.5:0.2:0.3) mixture. The peaks of ST-1 and ST-2 were not observed on each HPLC chromatogram at low sensitivity(2X), but by bioassay they were detected in the fraction of 24-27ml(less polar toxin, 120ng) and 22-27 ml (polar toxin, 150 ng). Less polar ciguatoxin from morey eel viscera also showed its peak in the same elution volume(25ml). Being subjected to chromatography on basic aluminum oxide (activity grade I) or to alkaline treatment, followed by basic aluminum oxide (activity grade I) chromatography ST-1 toxin was remarkably converted into the polar toxic component supposed to be polar ciguatoxin in both cases. In the latter case, approximately 74% of the residual toxicity was changed into the polar component, accompanied by about 50% loss of the initial toxicity. More than 26% of ST-2 toxicity was transformed into the less polar toxic component supposed to be less polar ciguatoxin on a deactivated aluminum oxide (activity grade V) column.

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Effects of Paper Sludge Application on the Chemical Properties of Paddy Soil and Growth of Paddy Rice II. Effects of Paper Sludge Application on the Seasonal Variations of Humus in Paddy Soil (제지(製紙)슬러지의 시용(施用)이 논 토양(土壤)의 화학성(化學性)과 수도생육(水稻生育)에 미치는 영향(影響) II. 토양중(土壤中) 부식형태(腐植形態)에 미치는 슬러지의 영향(影響))

  • Heo, Jong-Soo;Kim, Kwang-Sik
    • Korean Journal of Environmental Agriculture
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    • v.5 no.1
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    • pp.1-10
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    • 1986
  • To investigate the effects of paper sludge on the seasonal variations of soil humus, paper sludges were applied to the pots at the rates of 600㎏/10a which was either preadjusted C/N ratio to 30 : 1 or not adjusted. The effects were compared with those of control. 1) The contents of ether soluble materials, resins, water soluble polysaccharides, hemicellulose, cellulose, ligno-protein, humic acid and fulvic acid were higher in the sludge treated soil than in the control, furthermore, the content of ligno-protein had positive correlation with that of organic nitrogen in soil. 2) Optical density of UV and visible spectra of humic acid obtained from all the treated soil was decreased with increasing wavelength. In functional groups of humic acid, phenolic-OH/alcoholic-OH ratio was slightly higher in the sludge treated soil than in the control. The types of humic acid in all treated soil were P and Rp types. 3) The infrared spectra of humic acid extracted from the soil were characterized by main absorption bands in the regions of $3, 400cm^{-1}$(H-bonded OH), $2,900cm^{-1}$ (aliphatic C-H stretching), $1,630cm^{-1}$ (aromatic C=C and/or H-bonded C=O) and $1,050cm^{-1}$ (Si-O of silicate impurity).

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Antioxidant Activity of Different Parts of Lespedeza bicolor and Isolation of Antioxidant Compound (싸리나무(Lespedeza bicolor) 부위별 추출물의 항산화 활성 및 항산화물질 분리)

  • Lee, Jae-Hak;Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.763-771
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    • 2012
  • In this study, total antioxidant properties of extracts from different parts of Lespedeza bicolor were determined using techniques of measuring 1,1-diphenyl-2-picryl hydrazyl/2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)-radical scavenging activity and total phenolic contents. The total antioxidant activities of leaf, stem and root extracts from various solvents (water, 50, 70, 100% ethanol, and hot-water) indicated that 50 and 70% ethanol extracts have high radical scavenging activities and phenolic contents. A systematic approach was used to determine the total antioxidant activity of different solvent fractions of the Lespedeza bicolor extracts, partitioning with chloroform, ethyl acetate, n-butanol, and water, and the ethyl acetate fraction was found to have the strongest antioxidant activity. Antioxidant assay-guided isolation was carried out to isolate potential antioxidant compounds. The ethyl acetate fraction of the leaf extract was subjected to silica gel, LH-20 and RP-18 column chromatography successively, and afforded compound 1, which was identified as eriodictyol by NMR and MS analysis, after which its antioxidant activity was determined.