• Title/Summary/Keyword: Skin concentration

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Phototoxicity Evaluation of Pharmaceutical Substances with a Reactive Oxygen Species Assay Using Ultraviolet A

  • Lee, Yong Sun;Yi, Jung-Sun;Lim, Hye Rim;Kim, Tae Sung;Ahn, Il Young;Ko, Kyungyuk;Kim, JooHwan;Park, Hye-Kyung;Sohn, Soo Jung;Lee, Jong Kwon
    • Toxicological Research
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    • v.33 no.1
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    • pp.43-48
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    • 2017
  • With ultraviolet and visible light exposure, some pharmaceutical substances applied systemically or topically may cause phototoxic skin irritation. The major factor in phototoxicity is the generation of reactive oxygen species (ROS) such as singlet oxygen and superoxide anion that cause oxidative damage to DNA, lipids and proteins. Thus, measuring the generation of ROS can predict the phototoxic potential of a given substance indirectly. For this reason, a standard ROS assay (ROS assay) was developed and validated and provides an alternative method for phototoxicity evaluation. However, negative substances are over-predicted by the assay. Except for ultraviolet A (UVA), other UV ranges are not a major factor in causing phototoxicity and may lead to incorrect labeling of some non-phototoxic substances as being phototoxic in the ROS assay when using a solar simulator. A UVA stimulator is also widely used to evaluate phototoxicity in various test substances. Consequently, we identified the applicability of a UVA simulator to the ROS assay for photoreactivity. In this study, we tested 60 pharmaceutical substances including 50 phototoxins and 10 non-phototoxins to predict their phototoxic potential via the ROS assay with a UVA simulator. Following the ROS protocol, all test substances were dissolved in dimethyl sulfoxide or sodium phosphate buffer. The final concentration of the test solutions in the reaction mixture was 20 to $200{\mu}M$. The exposure was with $2.0{\sim}2.2mW/cm^2$ irradiance and optimization for a relevant dose of UVA was performed. The generation of ROS was compared before and after UVA exposure and was measured by a microplate spectrophotometer. Sensitivity and specificity values were 85.7% and 100.0% respectively, and the accuracy was 88.1%. From this analysis, the ROS assay with a UVA simulator is suitable for testing the photoreactivity and estimating the phototoxic potential of various test pharmaceutical substances.

Anti-inflammatory effects of the Rubus occidentalis seed extracts on UV-B induced inflammation in HaCat cells (각질형성세포에서 UV-B와 사이토카인에 의하여 유도되는 염증 반응에 대한 복분자 씨앗 추출물의 항염증 효과)

  • Kim, Dong-Hee;Park, Tae-Soon;Son, Jun-Ho
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.247-253
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    • 2016
  • It confirmed the applicability as an anti-inflammatory material from Rubus occidentalis seed (RSE) extract. In HaCaT cells to evaluate the anti-inflammatory potential as a material RSE extract on the activity of the inflammatory factors caused by UVB and $IFN-{\gamma}/TNF-{\alpha}$. We measured the activity of ROS, interleukin-$1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6) and interleukin-8 (IL-8) by ROS-Glo $H_2O_2$ assay and ELISA kit. Our results showed that the RSE extracts inhibit the UVB and $IFN-{\gamma}/TNF-{\alpha}$-induced ROS activities and expression of $IL-1{\beta}$, IL-6 and IL-8 in a dose-dependent manner. Also it was found that inflammatory mediators of the expression of cyclooxygenase-2 (COX-2) inhibition were also brought, the expression of which is increased $PGE_2$ by COX-2 also inhibited. Finally RSE extracts measure the seed expression of filaggrin in the skin barrier, the main factor of the extract could be confirmed to increase the expression of the filaggrin damaged as a result of this concentration-dependent manner. Through this, it was able to confirm that the efficacy RSE extract to protect the inflammation by restoring the damaged layers of the epidermis. Results from more than RSE extract was able to confirm that the extract that has anti-inflammatory effects by improving the inflammation being produced from UVB.

Airborne Bacteria Concentration and Species Identification in Residential Living Spaces (주택내 주거공간에 따른 부유세균 농도 분포 및 종 동정 연구)

  • Kim, Sung-Yeon;Jheong, Weonhwa;Hwang, Eun-Seol;Kim, Ji-Hye;Jung, Joon-Sig;Lee, Jae-won;Chung, Hyen-Mi;Kwon, Myunghee
    • Journal of Environmental Health Sciences
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    • v.42 no.6
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    • pp.438-449
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    • 2016
  • Objectives: Exposure to airborne bacteria is associated with adverse health effects such as respiratory and infectious diseases. This study evaluated airborne bacterial concentrations in the living rooms, kitchens, and toilets of 30 homes. Methods: Bacteria were sampled with an MAS100 impactor in three spaces in the subject homes between April 2014 and February 2015. Bacteria were grown on TSA plates for 48 hours at $35^{\circ}C$. The bacterial strains were isolated and amplified by polymerase chain reaction. Results: The most culturable bacteria were found in toilets ($624.0CFU/m^3$, GM: $417.3CFU/m^3$), followed by in the kitchen ($503.8CFU/m^3$, GM: $324.9CFU/m^3$). The dominant genera identified were: Staphylococcus sp.(19%), Micrococcus sp.(16%), and Bacillus sp.(11%) in the indoor air and Bacillus sp. (30%) in the outdoor air. Gram-positive bacteria comprised more than half of all colonies. Conclusion: In this study, culturable bacteria concentrations were higher than those reported in other spaces. Therefore, it is important to control relative humidity and remove moisture to prevent bacteria from multiplying. Additionally, the dominant species in indoor air were Staphylococcus sp. and Micrococcus sp. These are found on the human skin, mucous membranes, and hair, so human activity can affect bacterial distribution. Therefore, cleaning and controlling moisture are important for reducing indoor bacterial concentrations.

Screening of Hyaluronidase Inhibitor in Korean Medicinal Plants (천연물로부터 히알루로니다아제 저해제 검색)

  • Hwang, Seon Gu;Yang, Anna;Kim, Soo Jung;Kim, Min Kee;Kim, Sung Soo;Oh, Hyun Jung;Lee, Jung Dae;Lee, Eun Ju;Nam, Kung-Woo;Han, Man-Deuk
    • Journal of Life Science
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    • v.24 no.5
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    • pp.498-504
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    • 2014
  • Mammalian hyaluronidases (HAase, EC 3.2.1.35) are a family of enzymes that hydrolyse N-acetyl-D-glucosamine (1-4) glycosidic bonds in hyaluronic acid, which is found in skin, cartilage, and the vitreous body. Although HAase is generally present in an inactive form within subcellular lysosomes, it is released in an active form in some types of inflammation and tissue injuries, thereby contributing to the inflammatory response. The HAase inhibitory activity of 500 methanolic extracts of 500 species from medicinal plants was screened using a Morgan microplate assay. The viscosity of the hyaluronic acid was measured with an Ubbelohde viscometer. Three MeOH extracts inhibited more than 50% of HAase activity at a concentration of 2 mg/ml. HAase inhibitory rates (%) of three species of medicinal plant extracts, Styrax japonica, Deutzia coreana, and Osmanthus insularis were 57.28%, 53.50%, and 53.19%, respectively. The rate of HAase inhibition of the extracts was dose dependent. In the HAase inhibitory assay using the Ubbelohde viscometer, the results were in good agreement with the results from the Morgan assay. The results suggest that HAase inhibitory compounds extracted from the stem of S. japonica, D. coreana, and O. insularis might be multifunctional and prevent the degradation of hyaluronic acid and the induction of allergic reactions and inflammation.

Screening of Cytotoxicity and Antimicrobial Effects of Extracts from Atractylodes macrocephala Koidz (백출 추출물의 세포독성과 항균효과검색)

  • Choi Eun Young;Oh Hyun Ju;Park Nang Kyu;Chun Hyun Ja;Ahn Jong Woog;Jeon Byung Hun;Han Du Seok;Lee Hyun Ok;Baek Seung Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.348-352
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    • 2002
  • This study was carried out to evaluate cytotoxic effects of Atractylodes macrocephala Koidz. (A. macrocephala Koidz.) extract on NIH 3T3 fibloblast. SK-MEL-3 (HBT 69) and KB (ATCC No, OCL 17) cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay. 10.2 mg/ml Concentration of A. macrocephala Koidz. extracts in SK-MEL-3 showed that their susceptibility (sensitivity) to these compounds decreased in the following order ; adriamycin > H₂O > ethyl acetate > ethyl alcohol > chloroform > n-hexane in SK-MEL-3 cell lines ; 5-FU > H₂O > n-hexane > ethyl acetate > ethyl alcohol > chloroform in KB cell lines. In order to develop an antimicrobial agent, A. macrocephala Koidz. was extracted with solvents. The minimal inhibitory concentrations (MICs) of each solvent extract of A. macrocephala Koidz. against microogranisms were also examined. Antimicrobial activities of ampicillin and ketoconazole as references were compared to those of each solvent extract of A. macrocephala Koidz. The antimicrobial activity of the ethyl acetate soluble extract of A. macrocephala Koidz. had growth inhibition activity against S. mutans and P. putida (MICs. 500 ㎍/ml). These results suggest that the ethyl acetate soluble extract of A. macrocephala Koidz. possessed antitumorous and antimicrobial agents

Cellular Protective Effect and Component Analysis of Euphorbia humifusa Extracts (땅빈대 추출물의 세포 보호 효과 및 성분 분석에 관한 연구)

  • Kim, Sun-Young;Won, Doo-Hyun;Lim, Myoung-Sun;Park, Soo-Nam
    • Korean Journal of Pharmacognosy
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    • v.41 no.4
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    • pp.264-269
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    • 2010
  • In this study, the cellular protective effect, antioxidative property and component analysis of Euphorbia humifusa extracts were investigated. The ethyl acetate fraction ($3.68\;{\mu}g/mL$) and aglycone fraction ($3.15\;{\mu}g/mL$) of Euphorbia humifusa extract showed prominent free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$). Reactive oxygen species (ROS) scavenging activity ($OSC_{50}$) of Euphorbia humifusa extract on ROS generated in $Fe^{3+}$-EDTA/$H_2O_2$ system was investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction ($0.43\;{\mu}g/mL$) and aglycone fraction ($0.35\;{\mu}g/mL$) of extract showed higher ROS scavenging activity than L-ascorbic acid ($1.50\;{\mu}g/mL$). The cellular protective effects of fractions of Euphorbia humifusa extract on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fraction and aglycone fraction of extract protected cellular membranes against ROS in a concentration dependent manner ($5{\sim}25\;{\mu}g/mL$), and was more effective than (+)-${\alpha}$-tocopherol, lipid peroxidation chain blocker. Aglycone fraction from Euphorbia humifusa extract showed 2 bands in TLC and 2 peaks in HPLC. In HPLC chromatogram of aglycone fraction, peak 1 and peak 2 were identified as quercetin and kaempferol, respectively. And these components are very effective as antioxidant. Thus, these results indicate that fractions of Euphorbia humifusa extracts can function as antioxidant in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Fractions of Euphorbia humifusa extracts can be applicable to new functional cosmetics for antioxidant.

Effects on health by sleeping habits from few local university students (지역사회 일부 대학생의 수면습관이 건강에 미치는 영향 요인)

  • Seo, Soo-Gyo;Kim, Sung-Woo
    • The Journal of Korean Society for School & Community Health Education
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    • v.8 no.2
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    • pp.61-77
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    • 2007
  • Background & Objective, Methods: This study aims to analyze the reason why if university students feel lack of sleep and study how much we know about correct sleeping habits and thus increase attention on correct sleeping habits and prepare plans for applying this into our lives thus 210 male and female students from 2 universities in Daegu were studied during May 10 and 15 of 2006 and the summary and results are as follows. Results: Sleeping is very important since it as much as takes about 1/3 of our lives. However, most of students do not sleep well. Therefore, this study researched the reason for not sleeping well and the status on incorrect sleeping habits subjecting 210 university students. The results of this research showed that average sleeping hours are for $6{\sim}8$ hours and majority of students went to bed before lam and got up before 7am. Majority of students went to bed late to watch TV or doing computers from both male and female students followed by attending various gatherings. It took $10{\sim}20$ minutes to fall asleep and reasons for waking up while sleeping was with no special reason. The most sleepy time was after meals at 39% followed by during class, during daytime, while using transportations. More female students answered to take nap than male students and majority of them answered as taking nap for less than 2 hours. And, 51% answered that they slept well and 49% answered that they did not, and majority of them answered the reason for not sleeping well as being sensitive, or because of dreams and nightmares. Chronic fatigue was the biggest cause for lack of sleeping followed by fall in concentration, skin diseases. Conclusions: The results of this study showed that we need to take plenty of sleep by exercising regularly or sleeping for daily appropriate hours, maintaining peaceful mind before going to bed, avoid taking naps and limiting the time for watching TV and doing computers to develop correct sleeping habits. Also, we hope that it will Provide an opportunity to live healthy and quality lives mentally as well as physically by realizing and practicing the importance and necessity of sleeping.

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Whitening and Anti-wrinkle Effects of Prunus persica Flos (도화(桃花, Prunus persica Flos)의 미백 및 주름개선 효과)

  • Lee, Jin-Young;An, Bong-Jeun
    • Journal of Applied Biological Chemistry
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    • v.53 no.3
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    • pp.154-161
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    • 2010
  • The solvent extracts of Prunus persica Flos were investigated for the activities of whitening and anti-wrinkle effects to apply as a functional ingredient for cosmetic products. The tyrosinase inhibitory effect, which is related to skin-whitening, was 54.0, 58.3% in P. persica Flos (PPW, PPE) at 1,000 ppm. In addition, the ethanol extract of P. persica Flos (PPE) showed a potent tyrosinase inhibitory activity in the test using melanoma cell lines resulting in 40.0% inhibition at 100ppm. Furthermore, the aqueous acetone extract from the flower of P. persica Flos was found to inhibit elastase, which was more effective than ascorbic acid at 1,000 ppm. The inhibition of melanin synthesis by P. persica Flos extract (PPE) was about 56.5% at 100 ppm concentration. When compared to other extraction methods, the ethanol extract showed more potent whitening activity. For anti-wrinkle effect, the elastase inhibition activity of P. persica Flos extract (PPA) was 57.0% and higher than that of ascorbic acid at 1,000 ppm. The collagenase inhibition activity of P. persica Flos extract (PPA) was about 48.0% at 1,000 ppm. Collagen synthesis in fibroblast cell by P. persica Flos extracts (PPA) was about 41.0% at 100 ppm and its acetone extract was the best showing antiwrinkle activities. All these findings suggested that P. persica Flos has a great potential as a cosmeceutical ingredient with a whitening and anti-wrinkle effect.

Cytotoxity and Antimicrobial Effects of the Extract of Poncirus trifoliata (지실 추출물의 세포독성과 항균효과)

  • Yang Hyun-Ok;Oh Hyun-Ju;Park Nang-Kyu;Choi Eun-Young;Lee Hyun-Ok;Yang Eun-Yeong;Chun Hyun-Ja;Chung Soon-Ryang;Lee Kang-Min;Baek Seung-Hwa
    • Journal of Society of Preventive Korean Medicine
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    • v.4 no.2
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    • pp.235-241
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    • 2000
  • This study was carried out to evaluate cytotoxic effects of Poncirus trifoliata Raf. extract on lymphocytic leukemia tumor (L1210) cell lines. Disruptions in cell organelles were determined by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT) assay The comparison of Ic50 Values of Poncirus trifoliata Raf. extract in L1210 cell lines showed that their susceptibility to these fractons decreased in the following order: adriamycin > Fr.4> Fr. 6> Fr. 5> Fr. 3> Fr. 1> Fr. 2 by the MTT assay. In order to develop an antumicrobial agent, Poncirus trifoliata Raf. was extracted wit ethanol, and then it was fractionated with several mobile phase. The antitumor activities of fractions of the ethanol soluble extract was investigated. The minimal inhibitory concentrations (MIC) of fractions of the ethanol soluble extract of Poncirus trifoliata Raf. against microorganisms were also examined. Antimicrobial activities of ampicillin and ketoconazole as references were compared to those of fractions of the ethanol soluble extract of Poncirus trifoliata Raf. The antimicrobial activities of all fractions from the extract had growth inhibition activities against gram-positive bacteria, gram-negative bacteria and fungi $(MIC\;>\;200{\mu}g/ml)$. These results suggest that fraction 4 of the ethanol soluble extract of Poncirus trifoliata Raf. possessed the most antitumorous agent.

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Effects of Sulfobutyl Ether $\beta$-Cyclodextrin on Physicochemical Properties of Dexamethasone Dipropionate

  • Moon, Jee-Hyun;Oh, Ik-Sang;Chun, In-Koo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1997.04a
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    • pp.116-116
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    • 1997
  • Complex formation of practically insoluble dexamethasone dipropionate (DDP) with ${\beta}$-cyclodextrin (${\beta}$-CD), dimethyl-${\beta}$-cyclodextrin (DMCD), trimethyl-${\beta}$-cyclodextrin (TMCD), 2-hydroxypropyl-${\beta}$-cyclodextrin (HPCD) and sulfobutyl ether ${\beta}$-cyclodextrin (SBCD) in water was investigated by solubility method at various temperatures. Water solubility of DDP was found to be 1.78 $\mu\textrm{g}$/$m\ell$ at 37$^{\circ}C$. Propylene glycol (PG)-water cosolvent increased the solubility of DDP, but the solubilization was not sufficient (8.93 $\mu\textrm{g}$/$m\ell$ in 20% PG). The addition of CD markedly increased the solubility of DDP in water, and A$\sub$L/ type phase solubility diagrams were obtained with ${\beta}$-CD, TMCD, HPCD and SBCD, where the apparent stability constants of the soluble complexes at 25$^{\circ}C$ were determined to be 1388, 216, 1054, and 1992 M$\^$-1/, respectively. However, DMCD remarkably increased the solubility of DDP, and showed an A$\sub$P/ type diagram, suggesting that DMCD forms a soluble complex of high order with DDP. The stability constant for the DDP-DMCD complex at 25$^{\circ}C$ was determined to be 19132 M$\^$-1/. The thermodynamic parameters were calculated for the inclusion complex formation in aqueous solution. CD (1${\times}$10$\^$-2/M) remarkably decreased the partition coefficients of DDP between isopropyl myristate/water in the order of TMCD < ${\beta}$-CD < HPCD < SBCD < DMCD, and in squalane/water system in the order of HPCD < TMCD < ${\beta}$-CD < DMCD < DMCD $\leq$ SBCD. This finding represents that, in a o/w type cream, cyclodextrin complexation with DDP may result in high concentration of DDP in aqueous phase. The permeation of DDP through a cellophane membrane was highly suppressed by the addition of CD, and the degree of suppression was different among CDs, indicating that CD may control the skin permeation of DDP. The dissolution rates of solid dispersions with CDs were much faster than those of drugs alone and corresponding physical mixtures. All DDP-CD solid dispersions exceeded the equilibrium solubility. Consequently these results suggest that complex formation of DDP with CDs may provide useful means to markedly enhance the solubility, and CDs are useful in the semi-solid preparations such as creams and gels for topical application.

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