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Isolation and characterization of sigH from Corynebacterium glutamicum (Corynebacterium glutamicum의 sigH 유전자의 분리 및 기능분석)

  • Kim Tae-Hyun;Kim Hyung-Joon;Park Joon-Sung;Kim Younhee;Lee Heung-Shick
    • Korean Journal of Microbiology
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    • v.41 no.2
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    • pp.99-104
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    • 2005
  • Corynebacterial clones which exert regulatory effects on the expression of the glyoxylate bypass genes were isolated using a reporter plasmid carrying the enteric lacZ fused to the aceB promoter of Corynebacterium glutamicum. Some clones carried common fragments as turned out by DNA mapping technique. Subcloning analysis followed by the measurement of $\beta-galactosidase$ activity in Escherichia coli identified the region responsible for the aceB-repressing activity. Sequence analysis of the DNA fragment identified two independent ORFs of ORF1 and ORF2. Among them, ORF2 was turned out to be responsible for the aceB-repressing activity. ORF1 encoded a 23,216 Da protein composed of 206 amino acids. Sequence similarity search indicated that the ORF may encode a ECF-type $\sigma$ factor and designated sigH. To identify the function of sigH, C. glutamicum sigH mutant was constructed by gene disruption technique and the sigH mutant showed growth retardation as compared to the wild type strain. In addition, the mutant strain showed sensitivity to oxidative-stress generating agent plumbagin. This result imply that sigH is probably involved in the stress response occurring during normal cell growth.

Quality Characteristics and Antioxidant Activities of Sulgidduk (Rice Cake) Added with Chia (Salvia hispanica L.) Seed Powder (치아시드 분말을 첨가한 설기떡의 품질 특성 및 항산화 활성)

  • O, Hyeonbin;Choi, Byung Bum;Kim, Young-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.1
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    • pp.61-67
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    • 2017
  • Chia seed (Salvia hispanica L.) originated from Central America is a highly nutritious food containing large amounts of linolenic acid, dietary fiber, and protein. This study investigated the quality properties and antioxidant activities of Sulgidduk prepared with chia seed powder as a functional material. Freeze-dried chia seed powder was replaced with 0, 1, 3, 5, and 7% of rice flour. The addition of chia seed powder did not affect water content, whereas the pH value of the chia seed group decreased as compared to the control. For color measurements, a and b values increased as the amount of chia seed powder increased, whereas L value decreased. Hardness and chewiness of Sulgidduk with chia seed powder were lower than those of the control, whereas springiness of the chia seed group was higher than that of the control. Cohesiveness was not significantly different in all samples. According to retrogradation analysis based on changes in hardness during storage, it was confirmed that addition of chia seed powder inhibited aging of Sulgidduk. Retrogradation of CSP5 was the slowest. Consumer acceptability analysis did not show significant differences in all samples. Total polyphenol and flavonoid contents tended to significantly increase as chia seed content increased. DPPH and ABTS radical scavenging activities of Sulgidduk were also elevated due to addition of chia seeds. From the results, addition of chia seed softened texture, inhibited aging, and enhanced antioxidant activities of Sulgidduk. It is concluded that addition of 5% chia seed powder, which showed high effectiveness for aging, is the most suitable for commercialization.

Effect of the magnetism(neodymium magnet) on growth factor receptors of osteoblasts (희토류 자석의 자성이 골모세포 성장인자 수용체의 증가에 미치는 영향에 관한 연구)

  • Lee, Sang-Min;Lee, Sung-Bok;Choi, Boo-Byung
    • Journal of Dental Rehabilitation and Applied Science
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    • v.19 no.2
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    • pp.87-96
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    • 2003
  • The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

Quality Characteristics of Doenjang by Aging Period (전통 된장의 숙성 기간에 따른 감각·화학적 품질특성)

  • Ku, Kyung-Hyung;Park, Kyungmin;Kim, Hyun Jung;Kim, Yoonsook;Koo, Minseon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.5
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    • pp.720-728
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    • 2014
  • In order to characterize the quality of Doenjang, fermented Korean soybean paste, subjected to long-term aging, this study performed physico-chemical analyses and sensory evaluation according to aging period (from 1 to 9 years). Regarding the proximate composition of Doenjang according to aging period, moisture, crude protein, crude lipid, crude ash, and salt contents showed little differences among Doenjang samples. Amino-type nitrogen content was 1,046.7 mg% in the 1 year-aged sample, 990.9~996.9 mg% in the 2~5 year-aged samples, and 1,214.1~1,304.8 mg% in the samples fermented more than 5 years. ${\Delta}$E value, reflecting total color differences between the samples, increased according to aging period. Ratios of linoleic and linolenic acids, which are essential fatty acids in soybeans, constituted 55% of total fatty acids, which was the most abundant among all fatty acids. The major free sugar in Doenjang was fructose at a content of 1.6~2.2% in 1~9 year-aged Doenjang. Glycoside form of isoflavones in Meju constituted 77.1%, and the aglycon form constituted 22.9%. However, the glycoside type of isoflavones in soybeans was converted to aglycon type in Doenjang through fermentation and aging. In the sensory evaluation of Doenjang samples, brown color, salt smell, soy sauce flavor, and viscosity all increased according to aging period, whereas sweet flavor, roast smell, beany flavor, salty taste, and acrid taste showed no significant differences. In cluster analysis of the sensory attributes of Doenjang according to aging period, 1 year-aged Doenjang was significantly different between 2 year- and 3~5 year-aged Doenjang.

Changes of Physicochemical Properties of Conjugated Linoleic Acid(CLA) Accumulated Pork during Storage Time (Conjugated Linoleic Acid(CLA)가 축적된 돈육의 저장기간중 이화학적 특성변화)

  • 이정일;정재두;이진우;이제룡;정진연;양한술;주선태
    • Food Science of Animal Resources
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    • v.23 no.3
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    • pp.236-249
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    • 2003
  • To investigate the effects of dietary conjugated linoleic acid 011 changes of physicochemical properties of pork. Twenty pigs were divided into 5 treatment groups and subjected to each experimental diet(0, 1.25% and 2.5% CLA for 2 weeks, 1.25% and 2.5% CLA for 4 weeks, as total fed diet before slaughtering(about 110 kg). Pork loin samples were aerobically packed and stored for 14 days at 4$^{\circ}C$. Samples were then analyzed for general compositions, physicochemical characteristics and sensory evaluation. pH value of CLA fed group pork was significantly increased than that of control group(p<0.05). Crude fat content of CLA treated group pork was significantly higher than the control pork(p<0.05), but there were no significant differences in crude protein, crude ash and total moisture contents between control and CLA treated groups. AU CLA fed group pork showed higher WHC value than control pork. Drip loss was significantly lower compared to those of control porks. No significant differences in lean meat and fat color(CIE L*, a*, b*) were observed between control and the CLA treated group porks. Meanwhile, CLA pork tended to be lower in content of total heme pigment compared to control pork. No remarkable differences were found in sensory properties(color and drip loss) among control and CLA diet-fed group porks. Marbling score and acceptability were significantly increased in CLA diet-fed pork compared to the control pork.

STUDIES ON THE DIMORPHISM AND TRANSITION OF BISEXUALITY OF HETEROSTYLOUS POLYGONACEAE (여뀌과 이형경식물의 Dinorphism과 Bisexuality의 변화)

  • Harn, Chang-Yawl
    • Journal of Plant Biology
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    • v.3 no.2
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    • pp.6-18
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    • 1960
  • The present experiments were designed in order to clarify the differences between the long and short styled plants and the transgressive gradition in the degree of dimorphism among the three heterostylous species of the Polygonus, P. japonica, F. esculentum, and P. senticosa, based on investigations regarding the floral structure, ecological and physiological traits, the results of which are summarized as follows: (1) P. japonica, although it exhibits typical dimorphism, has undergone so high a differentiation between long and short styled that its long styled individuals behave as if they were female; and short styled individuals as if male. In long-styled individuals, filament, anther, and pollen grains show signs of degeneration, most of the pollen being abortive. On the other hand, in short styled individuals, the filament, anther, and pollen grains have attained remarkable development; the pollen grians are large and fertile. In short-plant the fertilized flowers readily drop off in every stage of their embryo development. This species has completely lost the self-fertile property, which is characteristic of the non-dimorphic Polygonum genus. Although this specsei typically exhibits the physiological characteristics of the non-dimorphic Polygonum genus. Although this specisei typically exhibits the physiological characteristics of dimorphism in controlled pollination, the short-styled individuals bear no seed in nature, thus misleading taxonomists to idenfity the short-styled plant as male. 2) The morphological feature of the flower organ of P. senticosa obviously indicates definite dimorphism. Physiologically, however, no differentiation towards dimorphism was observed, the species still retaining, both in long and short-individuals, the self-fertile property common to the Polygonum genus. Elaborate examinations revealed that regardless of the modes of pollination, both fertiization and seed setting flourish, no differentiation betwen legitimate and illegitimate unions being recognizable. This sort of physiological property has not been observed in the investigations of other heterostylous plants. It is assumed that this species is differentiated structurally into dimorphism, but not yet physiologically. In nature, however, this plant would have more opportunities to be cross-pollinated, i.e., legitimately combined, than self-pollinated because of the development of two forms of flowers. 3) In terms of heterostylism, the F. esculentum just occupies the intermediate position between P. japonica and P. senticosa structurally, ecologically, and physiologically. Doescription of some of the physiological behavior of the plant will suffice to demonstrate the above facts. While P. japonica has completely lost its self-fertile property, P. senticosa still retains it wolly. In F. esculentum 2-6% of self-fertility is the result in illegitimate combination. There occur occasionally hereditary self fertile individuals among some of the F. or 20 min. irradiation plot, when they reach any stage of the same bacterial population. In addition to this increase of total population in the plots with the more dose of UV light irradiation, it seems that the more dose of UV light irradiation is the more shortened the generation time of Azotobacter. Therefore, it is clear that variation of reproductive rate must be, mere or less, due to the genetic effects induced by UV light irradiation. On the other hand, the lag phase or logarithmic growth phase in nonirradiated culture is shortened prominently, and this must be due to the difference in bacterial number of the original inoculm. The generation time of Azotobacter is shortened by exogeneous treatment of nuclei acid derivatives, and the degree is greater in case of DNA derivatives than RNA dervatives. W.H. Price reported that the rate of ribose nucleic acid to protein in Staphylococcus muscae is proportional to the generation time: that is the faster the cell can form ribose nucleic acid, the more rapid its growth. This explains the shortening of generation time by exogeneous RNA derivatives in this work reasonably. On the other hand, it is well known that the desoxyribose nuclic acid content per cell is constant and independent of the generation time. A.D. Laren and W.N. Takahashi reported that the infectious RNA from TMV is 6 times as sensitive to inactivation by UV as it is in the form of intact virus, and that inactivation of infectious TMV involves onlu a local change on RNA chain. But, the effect of exogeneous DNA in this work suggests that irradiated living cell which cotain DNA bring about some change on DNA moleculs as well as RNA molecules. And if the mutagenic effects of UV take into consideration, it is very reasonable. Therefore, it is clear that the variation of the generation time by UV irradiation is, more or less, due to the genetic effects. Therefore, it seems that the shortness of the average lifewpan of Azotobacter by UV irradiation is resulted not only from the influence of the environmental conditions, but also from the variation of genetic factor of the individual.

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Effects of Medicinal Enzyme Powder on Intestinal Mobility, Lipid Level, and Blood Parameters of Loperamide-Induced Constipation in Rats (약선효소 분말식이가 Loperamide의 투여로 유발된 변비 쥐의 장 운동과 지질 함량 및 혈액학적 성분 변화에 미치는 영향)

  • Park, Chan Sung;Park, Kyung Soo;Kim, Mi Lim;Kong, Hyun Joo;Yang, Kyung Mi
    • Journal of Life Science
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    • v.23 no.2
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    • pp.228-236
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    • 2013
  • This study was aimed at investigating whether dietary therapy using medicinal enzyme powder is effective in reducing constipation caused by loperamide in rats. Nine-week-old male Sprague Dawley were subdivided into 4 groups: normal diet group (C), loperamide treatment and normal diet (CL), medicinal enzyme powder diet (E), and loperamide treatment and medicinal enzyme powder diet (EL). Constipation was induced by subcutaneous injection of loperamide (1.5 mg/kg) 3 days prior to sacrifice. The treatment with loperamide led to an increase in weight gain, a decrease in the number and wet weight of fecal pellets, and a decrease in intestinal motility. The administration of the medicinal enzyme powder significantly reduced weight gain but increased intestinal mobility compared with the loperamide-treated group. The treatment with loperamide in the normal diet group reduced the activities of both suggesting that constipation may be involved in the low level of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT). Additionally, the loperamide treatment in the medicinal enzyme powder diet group increased the level of GOT, but reduced the level of GPT. Loperamide treatment also reduced cholesterol and increased the atherogenic index (AI) and cardiac risk factors (CRFs). Interestingly, the treatment with the medicinal enzyme powder effectively attenuated both the increase in AI and the reduction in high density lipopretein (HDL)-cholesterol, caused by the treatment with loperamide. Although there were no significant differences in the blood protein level, including hemoglobin and hematocrit, between the normal diet group and the loperamide-treated group, the administration of the medicinal enzyme powder to the loperamide-treated group effectively increased the levels of both hemoglobin and hematocrit. Collectively, the results demonstrate that the medicinal enzyme powder can help to combat the negative events caused by constipation.

Effects of Pig Slurry Application on Body Weight Gain of Grazing Hanwoo Heifer and Forage Yields in a Grass-Legume Mixed Pasture (방목초지 돈분액비 시용이 목초 생산성 및 방목한우 증체에 미치는 영향)

  • Ko, S.B.;Park, N.G.;Hwang, K.J.;Lee, C.E.;Kang, S.Y.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.23 no.4
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    • pp.255-264
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    • 2003
  • A study was conducted to determine the effects of pig slurry application on forage yield and growth of fazing Hanwoo heifer in a mixed pasture. To each of three treatments 1.4ha were alloted a control applied with only chemical fertilizer (N-P-K=150-150-120kg/ha), two pig slurry lots applied with the amount to allow 100% (150 kg/ha) or 150% (225 kg/ha) of N used in the control. A randomized block design was used without replication. Cattle were allowed to graze continuously during the experimental period. Results obtained were as follows: Total dry matter yield was 16,291, 15,632 and 16,320 kg/ha for chemical fertilizer. pig slurry 100% and 150%, respectively. The pasture was dominated by perenial ryegrass during the first gazing season, but by orchard grass and perenial ryegrass (60∼70%) and red clover (20∼30%) during the second grazing season. Average gazing rate per ha was 2.75∼2.76 animal units and daily weight gain of grazing cattle was not different among treatments ranging from 0.563 to 0.580 kg. Total weight gain of grazing cattle per ha during the grazing period was 541, 541 abd 555 kg for chemical fertilizer, pig -slurry 100 and 150%, respectively. RBC, WBC, total protein and albumin etc. concentrations in blood were normal in all treatments.

Studies on the Maturation of the Follicular Oocytes by Xenoplastical Transplantation in the Anterior Chamber of the Eye (異種動物의 眼前房에 이식된 濾胞卵子의 成熟에 관한 연구)

  • Cho, Wan Kyoo
    • The Korean Journal of Zoology
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    • v.13 no.3
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    • pp.68-74
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    • 1970
  • In the previous studies, the present author found that high proportion of the follicular oocytes from mouse and rabbit ovaries are able to resume their maturation division in the anterior chamber of the eye in which they have been incubated by auto- or homoplastic transplantation. Especially in the case of the homoplastic transplantation, it was known that no trouble has been detected in the process of resumption of the oval maturation in particular connection with the antigen-antibody reaction between donor and recipient. These findings provide a possibility that the follicular oocytes from various animals would be matured in the eye even after the xenoplastic transplantation. Under such an assumption, the present studies were performed to examine the behavior of the follicular oocytes in the eye chamber of the animals of different species. For the donor of the follicular oocytes, domestic rabbits, albino rats of Sprague-Dowley strain, and albino mice of A-strain bred in our laboratory were used. The oocytes obtained from the ovarian follicules were introduced to the anterior chamber of the eye of different species of animals, with an exception of rabbit in which only the female animals were used as a recipient. The procedures of collection of ova, introduction to the eye, harvest from the eye ball, fixation, and staining were the same as mentioned in the previous reports (Cho, 1967b; Cho and Kim, 1968). The conclusions obtained are summarized as below. 1. The rabbit follicular oocytes are able to mature in the eye chambers of both male mouse and rat, although the proportion of the maturation is lower than when they are incubated autoplastically in the eye. When the ova were incubated in the male mouse eye for 24 hours, 21 per cent of them showed chromosomes at metaphase I and II, whereas the rate was 32 per cent when they were incubated in the eye of the male rat. These are apparently low comparing to the rate of 52 per cent of autoplastic transplantation. 2. When rat follicular oocytes were transferred into the mouse eye chamber and recovered after 24 hours, 43 per cent of them produced the mataphase I and II chromosomes. This proportion was higher than the result of the homoplastic transplantation which yielded 23 per cent of the ova on maturation. 3. The most striking result was found in the experiment with mouse follicular oocytes. Seventy-six per cent of the oocytes resumed their maturation division within 24 hours after they were transferred into the male rat eye chamber, and this figure was significantly high compared to the result o 55 per cent obtained by the homoplastic transplantation. In the rat eye, the induction of the degenerative ova also was low (19%). On the other hand, the proportion of the oval maturation decreased to 45 per cent, while that of degeneration increased 33 per cent when they were incubated in the eye of the female rabbit. 4. It was apparent from the present experiments that the follicular oocytes can reveal their activation to maturation in the eye chamber which contains aqueous humor which is known to be composed of low protein content and of very little gamma-globulin which acts as an antibody(Oser, 1965), and that it shows higher osmolarity than blood serum(Levene, 1958). Taking these properities into consideration the humor may provide unfavourable environment to the cells and tissues incubated in. However, it could be noteworthy finding that only the follicular oocytes in the eye of the different species can grow in healthy condition although the maturation rates are varied with the animal species. The fact that the rabbit follicular oocytes show the lower proportion in maturation may be due to the greater amount of the yolk granules in the egg cytoplasm than those in the mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat mouse and rat oocytes. That the mouse oocytes incubated in the eye of the rat resumed their maturation process in greater proportion would e explained by the fact that the rat eye chamber particularly provides the better environment to the mouse oocytes than the eye chamber of mouse does.

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Study on the Anti-bacterium, Antioxidant and Anti-inflammatory Effect of Calystegia pubescens Lindl. Extracts (메꽃(Calystegia pubescens Lindl.) 추출물의 항균, 항산화 및 항염증 효과에 관한 연구)

  • Ji-A Byeon;Un-Gyo Shin;Ye-Jin Jang;Su-Bin Hwang;Seon-A Lee;Gayeon Kim;Jin-Tae Lee;Ildae Song;Yong-Jin Kwon
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.3
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    • pp.745-755
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    • 2024
  • Calystegia pubescens Lindl. (C. pubesens) is a native Korean herb that has been traditionally used for its diuretic, fatigue, and blood sugar-lowering effects. In previous studies, C. pubesens has shown antioxidant and whitening effects, but research on its antibacterial and anti-inflammatory properties is limited, and its potential as functional materials is lacking. In this study, we investigated the antibacterial, antioxidant, and anti-inflammatory effects of leaf (CPL) and stem (CPS) extracts of C. pubesens extracted with 70% ethanol. When the anti-bacterial effect was confirmed, CPL showed superior anti-bacterial effect than CPS for C. acne, S. aureus, E. coli and P. aeruginosa. Additionally, CPL exhibited significantly higher antioxidant activity compared to CPS, as demonstrated by DPPH and ABTS+ radical scavenging assays. Therefore, CPL showed superior anti-bacterial and antioxidant effects compared to CPS. Based on these results, further investigation was conducted to confirm the anti-inflammatory effect of CPL using RAW264.7 macrophages. Nitric oxide (NO) produced by lipopolysaccharide (LPS) stimulation was significantly reduced by CPL treatment in a dose-dependent manner. Additionally, the protein and mRNA expression of iNOS was reduced by CPL in LPS-indueced RAW264.7 cells, which was consistent with NO production. In conclusion, this study confirmed that CPL has superior anti-bacterial and antioxidant effects compared to CPS, and demonstrated anti-inflammatory effect of CPL via inhibiting iNOS expression and NO production induced by LPS. Based on the result, we suggest the potential of CPL as a valuable functional materials.