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Cloning of the β-Lactamase Gene from Bacillus sp. J105 and Analysis of Its Expression in E. colis Cells (Bacillus sp. J105 유래 β-lactamase 유전자의 cloning 및 E. coli 내에서의 발현 분석)

  • Kang, Won-Dae;Lim, Hak-Seo;Seo, Min-Jeong;Kim, Min-Jeong;Lee, Hye-Hyeon;Cho, Kyeong-Soon;Kang, Byoung-Won;Seo, Kwon-Il;Choi, Yung-Hyun;Jeong, Yong-Kee
    • Journal of Life Science
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    • v.18 no.11
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    • pp.1592-1599
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    • 2008
  • The $\beta$-lactamase gene was cloned into E. coli DH5$\alpha$ from Bacillus sp. J105 with strong resistance against $\beta$-lactam antibiotics. The chromosomal DNA was partially digested with Sau3AI and ligated to BamHI digested pLAFR3. $\beta$-Lactamase positive clones were obtained by using in vitro packaging kit. The pKL11-${\Delta}4.6$ with $\beta$-lactamase activity was obtained by subcloning of the recombinant plasmid ($\beta$-lac +). The 6.5 kb fragment in the subcloned plasmid was sequenced. The DNA fragment that contains the $\beta$-lactamase gene encodes 309 amino acids. The 0.17 kb upstream region was similar to those of B. thuringinesis and B. cereus with 97% identity. The deduced amino acids sequence was also similar to those of $\beta$-lactamase from B. thuringinesis and B. cereus with 97% and 94% identity, respectively. The phylogenetic tree also showed the relationships of the $\beta$-lactamase gene of Bacillus sp. J105 to genetically related that of other Bacillus strains. Analysis of expression pattern of the pKL11-${\Delta}4.6$ in E. coli, revealed that the secretion efficiency of $\beta$-lactamase was $4{\sim}5%$ and the molecular weight was as same as that of original $\beta$-lactamase (31 kDa) from Bacillus sp. J105.

Data Mining and Construction of Database Concerning Effects of Vitis Genus (산머루 관련 정보수집 및 데이터베이스의 구축)

  • Kim, Min-A;Jo, Yun-Ju;Shin, Jee-Young;Shin, Min-Kyu;Bae, Hyun-Su;Hong, Moo-Chang;Kim, Yang-Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.551-556
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    • 2012
  • The database for the oriental medicine had been existed in documentation in past times and it has been developed to the database type for random accesses in the information society. However, the aspects of the database are not so diversified and the database for the bio herbal material exists in widened type dictionary style. It is a situation that the database which handles the in-depth raw herbal medicines is not sufficient in its quantity and quality. Korean wild grape is a deciduous plant categorized into the Vitaceae and it was found experimentally that it has various medical effects. It is one of the medical materials with higher potentiality of academic study and commercialization recently because it has a bigger possibility to be applied into diverse industrial fields including the medical product for health, food and beauty. We constituted the cooperative system among the Muju cluster business group for Korean mountain wild grapes, Physiology Laboratory in Kyung Hee University Oriental Medicine and Medical Classics Laboratory in Kyung Hee University Oriental Medicine with a view to focusing on such potentiality and a database for Korean wild grapes was made a touchstone for establishing the in-depth database for the single bio medical materials. First of all, the literatures based on the North East Asia in ancient times had been categorized into the classical literature (Korean literature published by government organization, Korean classical literature, Chinese classical literature and classical literature fro Korean and Chinese oriental medicine) and modern literature (Modern literature for oriental medicine, modern literature for domestic and foreign herbal medicine) to cover the eastern and western research records and writings related to Korean wild grapes and the text-mining work has been performed through the cooperation system with the Medical Classics Laboratory in Kyung Hee University Oriental Medicine. First of all, the data for the experiment and theory for Korean wild grape were collected for the Medline database controlled by the Parliament Library of USA to arrange the domestic and foreign theses with topic for Korean wild grapes and the network hyperlink function and down load function were mounted for self-thesis searching function and active view based on the collected data. The thesis searching function provides various auxiliary functions and the searching is available according to the diverse searching/queries such as the name of sub species of Korean wild grape, the logical intersection index for the active ingredients, efficacy and elements. It was constituted for the researchers who design the Korean wild grape study to design of easier experiment. In addition, the data related to the patents for Korean wild grape which were collected from European Patent Office in response to the commercialization possibility and the system available for searching and view was established in the same viewpoint. Perl was used for the query programming and MS-SQL for database establishment and management in the designing of this database. Currently, the data is available for free use and the address is as follows. http://163.180.41.43:8011/index.html

Laboratory Diagnosis of Coronavirus Disease 19 (COVID-19) in Korea: Current Status, Limitation, and Challenges (국내 중증 급성 호흡기 증후군 코로나 바이러스의 검사실 내 진단: 현재, 한계점 그리고 직면한 과제)

  • Song, Gi Seon;Lee, You-Rim;Kim, Sungmin;Kim, Wontae;Choi, Jungwon;Yoo, Dahyeon;Yoo, Jungyoung;Jang, Kyung-Tae;Lee, Jaewang;Jun, Jin Hyun
    • Korean Journal of Clinical Laboratory Science
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    • v.52 no.3
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    • pp.284-295
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    • 2020
  • In December 2019, the first coronavirus disease- 2019 (COVID-19) patient was reported in Wuhan, Hubei Province, China. Since then, the number of patients who suffered severe acute respiratory syndrome caused by the novel Coronavirus (SARS-CoV-2 or 2019-nCoV) has increased dramatically in Korea. This new variant virus induces pulmonary diseases, including cough, sore throat, rhinorrhea, dyspnea, and pneumonia. Because SARS-CoV-2 is an RNA virus, real-time reverse-transcriptase PCR has been used widely to diagnose COVID-19. As the Korea Centers for Disease Prevention and Control (KCDC) and Ministry of Food & Drug Safety (MFDS) approved emergency use authorization, clinical specimens collected from COVID-19 patients and even healthy people have been clinically diagnosed by laboratory medicine. Based on a literature search, this paper reviews the epidemiology, symptoms, molecular diagnostics approved by KCDC, a current diagnosis of COVID-19 in the laboratories, the difference between molecular and serological diagnosis, and guidelines for clinical specimens. In addition, the Korean guidelines of biosafety for clinical laboratory scientists are evaluated to prevent healthcare-associated infection. The author's experience and lessons as clinical laboratory scientists will provide valuable insights to protect the domestic and international health community in this COVID-19 pandemic around the world.

The Effects of Cordyceps Militaris on Lipid Metabolism, Protein Levels and Enzyme Activities in Rats Fed a High Fat Diet (동충하초 (Cordyceps Militaris)가 고지방식이를 섭취한 흰쥐의 지질대사, 단백질농도 및 효소활성에 미치는 영향)

  • 고진복
    • Journal of Nutrition and Health
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    • v.35 no.4
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    • pp.414-420
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    • 2002
  • The effects of Cordyceps militaris on weight gain, flood intakes, flood efficiency ratios, serum and hepatic lipid concentrations, serum protein levels and serum enzyme activities, were studied in adult male rats. Sprague-Dawley rats, 35 weeks old, were given four different types of diets for a succeeding period of five weeks: either a normal diet(5% com oil), a high fat diet(5% com oil + 15% lard). a CF diet (high fat diet + 3% Suiting body of cordyceps militaris), or a CM diet (high fat diet + 3% mycelium of cordyceps military). The body weight gains, hepatic weights and food efficiency ratios of the rats find (high fat) diets with CF and CM were significantly decreased compared with those find the high fat diet, but were similar to those find the normal diet. The concentrations of serum and hepatic triglycerides, and hepatic total lipids and cholesterol, of rats given CF or CM diets were significantly lower than those given the high fat diet. But the concentrations of total cholesterol, HDL-cholesterol, phopholipid and total protein in the serum of rats find the high fat, CF or CM diets were significantly higher than those find the normal diet. In the serum of the rats find the CF diet, the HDL-cholesterol/total cholesterol ratio was significantly higher but the atherogenic index was significantly lower than those fed the high fat diet. The alkaline phosphatase activity in the serum of the rats find the high fat, CF, and CM diets was more significantly decreased compared to rats on the normal dirt. No differences were noted in the weights of the pancreas, kidney and heart, the serum concentrations of glucose, hemoglobin and albumin, and the activities of GOT, GPT and ${\gamma}$-GTP, among the rats on all the experimental diets. In conclusion, the rats find the Cf or CM diets maintained normal body and hepatic weights. Despite of the high intake of fats in the CF and CM diets, the concentrations of total lipids, cholesterol and triglycerides were decreased in the liver, also, the concentration of triglycerides was decreased in the serum.

Effects of Nutrient Intake and Exercise on Bone Mineral Density and Bone Mineral Density in Premenopausal Women (폐경 전 성인여성에서 영양섭취 상태와 운동이 골밀도 및 골무기질 함량에 미치는 영향)

  • 최미자
    • Journal of Nutrition and Health
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    • v.35 no.4
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    • pp.473-479
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    • 2002
  • This study investigated associations between calcium intake, exercise behaviors, lumbar bona mineral density (BMD), and bone mineral content (BMC) among 79 premenopausal women (mean age = 41yr). The BMD and BMC of the lumbar spine (L$_2$-L$_4$) were measured by dual energy x-ray absorptiometry. Nutrient intake was estimated by the convenient method and a quantitative food frequency questionnaire was designed for this study that included the most commonly consumed floods sources of Ca. Participants were asked to identify all activities of exercise they had participated in including estimation of number of years of participation, number of weeks per year, number of times per week, and the number of hours per session. Participants were then categorized into the exercise group or nonexercise group (control). To meet the criteria for inclusion in the exercise group, the subjects participated more than 3 sessions per week and more than 30 minutes per session and the length of the exercise participation was at least more then 6 months. The participants were also grouped by calcium intake. The total calcium intake of all participants was estimated by dietary calcium intake and then the subjects were divided into quartiles to assess the lumbar BMD and BMC of the upper 25% (average calcium intake = 910 mg) and the lower 25% (average calcium intake = 414 mg). Results indicated that there were no significant differences in energy and calcium intake, and that there were no significant differences in lumbar BMD and BMC between participants in exercise group and the nonexercising control group. However, the exercise group had significantly lower ALP concentration than the nonexercise group. The upper 25% calcium intake group had significantly greater lumbar bone mineral density and bone mineral content than the lower 25% calcium intake group. Also the upper 25% calcium intake group had significantly lower ALP concentration than the lower 25% calcium intake group. Correlation analysis revealed that the spinal BMB was positively associated with body weight, while calcium intake was negatively associated with ALP concentration in nonexercising women. However, neither body weight nor dietary calcium intake were associated with both spinal BMD or ALP concentration in exercising women. These results suggest that calcium intake positively influence bone mineral density and bone mineral content in nonexercisulg premenopausal women. Exercise group did not affected by body weight and dietary calcium, but decreased ALP concentration than nonexercising group. Both exercise and calcium intake positively influence bone mineral density and bone mineral content in premenopausal women.

Effects of Condiments upon ${\alpha}-Amylase$ activity (조미료(調味料)가 ${\alpha}-amylase$활성(活性)에 미치는 영향(影響))

  • Suh, Myung-Jah
    • Journal of Nutrition and Health
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    • v.9 no.1
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    • pp.104-109
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    • 1976
  • Condiments such as welsh onion, garlic, ginger, black pepper, red pepper, japanese pepper, mustard, horse radish, monosodium glutamate and sugar were ground by a homogenizer, and 0%, 1%, 5%, and 10% of each of the ground condiments were put into 0.2% ${\alpha}-amylase$ solution for storage at the temperature of $15^{\circ}C$. ${\alpha}-amylase$ activity then was measured by the Wohlgemuth method at 48-hour interval, and the following results were obtained. 1) Among the condiments, black pepper and sugar checked the ${\alpha}-amylase$ activity most, about 80% in comparison with control. 2) Welsh onion, garlic, mustard, and ginger checked the ${\alpha}-amylase$ activity, about 50% in comparison with control, irrespective of the time stored. 3) The low concentration of red pepper, horse radish, japanese pepper and mono-sodium glutamate checked the ${\alpha}-amylase$ activity about 50%, while the high one did below 75% in comparison with control. To conclude: all the condiments used in the experiment checked ${\alpha}-amylase$ activity.

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Comparative analysis on status of events and importance-performance analysis (IPA) between industry and school foodservices (사업체급식소와 학교급식소의 이벤트 실태 및 이벤트에 대한 중요도-수행도 분석 비교)

  • Song, Hyun Jin;Rho, Jeong Ok
    • Journal of Nutrition and Health
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    • v.47 no.6
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    • pp.452-462
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    • 2014
  • Purpose: The purpose of the study was to evaluate the status of events and importance-performance analysis (IPA) between industry and school foodservices. Methods: The study subjects were dietitians in industry foodservices (n = 73) and nutrition teachers in school foodservices (n = 135) in the Jeonbuk area. Demographic characteristics, status of events, and problems of implementing events were assessed using a self-administered questionnaire. Results: Approximately 67% of industry foodservices had implemented an event a month, whereas 40.7% of school foodservices did not implement events for students. The type of events that industry and school foodservices had implemented most frequently were Sambok event, Dongji event, Daeboreum event, Spring-Namul event, and Chuseok event. The industry foodservices had significantly higher average scores for performance of events than those of the school foodservices (p < 0.001). In the Importance-Performance Analysis (IPA), high importance and high performance (A area: doing great) in industry foodservices were seasonal events, traditional festival day events, anniversary events, traditional seasonal events, and personal memory events, whereas in school foodservices were traditional festival day events, traditional seasonal events, school events, and environment events. Conclusion: These results showed that events are important for the increase in customer satisfaction. Therefore, it is necessary to consider educational programs on event implementation for dietitians and employees in industry and school foodservices.

Survey on sanitary knowledge level and degree of HACCP practice in culinary staff employed in elementary, middle, and high schools in Chungnam province (충남지역 초.중.고 학교급식 조리원의 위생지식 및 HACCP 수행도 조사)

  • Lee, Young Joong;Kim, Sun Hyo
    • Journal of Nutrition and Health
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    • v.47 no.4
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    • pp.300-312
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    • 2014
  • Purpose: This study was performed to examine the sanitary knowledge level and degree of HACCP (hazard analysis critical control point) practice in school culinary staff in order to provide basic information for improving hygiene of school meals. Methods: Exactly 305 culinary staff members were selected from elementary, middle, and high schools in 14 cities and rural areas, including whole administrative districts in Chungnam province. Surveyed schools were selected by convenience sampling, and one subject was selected randomly from each school. Surveys were taken by self-administered questionnaires developed by researchers and questionnaire were distributed and collected by postal mail. Results: Sanitary education administered by school dietitians to culinary staff was more frequent and longer in elementary schools, followed by middle and high schools (p < 0.001). Sanitary knowledge level and degree of HACCP practice, except for a few CCP or CP of culinary staff, were highest in elementary schools and middle school followed by high schools (p < 0.05), respectively. School class was negatively correlated with sanitary knowledge level of culinary staff (p < 0.01), and frequency of sanitary education was positively correlated with sanitary knowledge level of culinary staff (p < 0.01). Sanitary knowledge level of culinary staff was positively correlated with degree of HACCP practice (p < 0.01). School class, daily frequency of meal service, work experience, rice washing machine, and total score of sanitary knowledge were significant variables influencing degree of HACCP practice in culinary staff. Conclusion: The above results show that the following points should be considered to improve hygiene of school meals. Sanitary education should be administered more frequently by school dietitians to culinary staff, especially to those in high schools, which showed the lowest sanitary knowledge level and degree of HACCP practice. In addition, facilities and equipment required for HACCP practice should be supported in small-sized elementary schools.

Biochemical and Nutritional Effects of Rice, Barley, and Millet on Rats (백미대맥(白米大麥) 및 속(粟) 사료(飼料)가 실험용(實驗用) 백서(白鼠)에 미치는 영양학적(營養學的) 및 생물화학적(生物化學的) 연구(硏究))

  • Haw, Kum
    • Journal of Nutrition and Health
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    • v.1 no.1
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    • pp.9-18
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    • 1968
  • Dividing into six(6) experimental groups; Stock Diet(SD) Group, Rice Diet(RD) Group, Rice Diet supplemented with Barley(RD-BD) Group, Barley Diet(BD) Group, Rice Diet supplemented with Millet(RD-MD) Group, and Millet Diet(MD) Group, biochemical and nutritional effects of rice, barley, and millet on rats are studied. The grains used in this experiment are polished ones and purchased from market. Six(6) weaning male albino rats are housed into the individual cages in each group and fed with each experimental diet and water ad libitum for 21 weeks. The composition of the experimental diets are shown in Table 1. At the end of experiment, the appearences of rats are observed and the bloods are sampled by heart puncture method following ether anesthesia for determinations of serum protein and serum cholesterol. And the livers are sampled immediately right after heart after heart puncture for determinations of liver weight, liver nitrogen, liver fat, and liver glycogen. And also liver tissues are observed histologically. The result of growth experiment is shown in Table 2 and Figure 7. The results of blood analysis are shown in Table 3 and the results of liver analysis are shown in Table 4. And the histological observations of liver tissue are shown in Figures 1, 2, 3, 4, 5, and 6. According to the above results, it could be summarized as follows; 1) The normal appearences are observed at SD Group and BD Group. Alopecia is observed at RD, RD-­BD, RD-MD, and MD groups and the wet appearences of hair are observed at RD-MD and MD groups. The color of hair is changed to yellowish white at RD, RD-MD groups. The maximum growth inhibition, is observed at MD group and the growths of RD, RD-BD, BD, and RD-MD groups are significantly lower than that of SD group. The growth of RD group is lower than those of RD-BD and BD groups, though this difference is not significant. 2) At the blood analysis, the highest serum protein content is shown at SD group and the lowest serum protein is observed at MD group. The serum cholesterol contents are not significantly different among the six(6) experimental groups. 3) The highest liver weight is observed at SD group and the lowest weight is shown at MD group. However, oppositely the highest liver weight per 100 grams of body weight at MD group and the lowest liver weight per 100 grams of body weight at SD group are observed as shown in Table 6. 4) The differences of liver nitrogen contents of RD, RB-BD, BD, RD-MD, and MD Groups are not significant except SD group showing the highest content. 5) The significant highest content of liver fat is observed at RD group and the lowest content of liver fat is shown at SD group. And no significant differences in contents of liver glycogen are observed among the six(6) experimental groups. 6) No significant abnormal histological changes of liver tissue are observed at SD, RD-BD, and BD groups. However, the fat metamorphosis at peripheral zone of liver tissue is observed at RD Group and the some histological abnormal changes are observed at RD-MD and MD groups. According to the above summaries, the followings might be estimated and recommended; 1) The BD group shows histological normality of liver tissue, improvement of liver fat and liver nitrogen, normality of appearence, and better growth compared with RD Group. This could be indicated that barley is nutritionally and biochemically better than rice as the feeding diet on rat. And it might be considered that there might be the species speciality for nutritionary evaluation of food. 2) Recently, Marshall indicated that the high consumption of carbohydrate causes fatty liver and Kondo reported that the higher consumption of rice causes the more heart disease and the shorter life span. Considering these reports, the higher consumption of rice as a main dish in our country should be considered as an important national nutritional problem. And this might be improved by supplementing with barley or other grains to rice.

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Suppressive effects of ethanol extract of Aralia elata on UVB-induced oxidative stress in human keratinocytes (자외선 B를 조사한 인간유래각질세포에서 두릅순 에탄올추출물의 산화적 스트레스 억제효과)

  • Kwak, Chung Shil;Yang, Jiwon
    • Journal of Nutrition and Health
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    • v.49 no.3
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    • pp.135-143
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    • 2016
  • Purpose: Ultraviolet (UV)-induced oxidative stress contributes to several adverse biological effects on skin. Many phenolic phytochemicals have been shown to have antioxidant properties and protect skin cells from UV-induced oxidative damage. In this study, we investigated whether or not Aralia elata (AE) has a protective effect against UVB-induced reactive oxygen species (ROS), ultimately leading to photoaging. Methods: Phenolic content of dried AE and antioxidant properties of AE extract in 70% ethanol weredetermined by measuring DPPH and ABTS radical scavenging activities and ferric reducing antioxidant power (FRAP). The effect of AE extract on cellular ROS generation and expression levels of oxidative stress-response proteins such as superoxide dismutase (SOD)-1, catalase, nuclear factor-erythroid 2-related factor (Nrf)-2, and heme oxygenase (HO)-1 in UVB-irradiated ($75mJ/cm^2$) human keratinocytes (HaCaT) were further determined by 2'-7'-dichlorofluoresceine diacetate assay and Western blotting, respectively. Results: The total phenolic and flavonoid contents of dried AE were 20.15 mg tannic acid/g and 18.75 mg rutin/g, respectively. The $IC_{50}$ of AE extract against DPPH radical was $98.5{\mu}g/mL$, and ABTS radical scavenging activity and FRAP upon treatment with $1,000{\mu}g/mL$ of AE extract were $41.8{\mu}g\;ascorbic\;acid\;(AA)\;eq./mL$ and $29.7{\mu}g\;AA\;eq./mL$,m respectively. Pretreatment with AE extract significantly reduced (p < 0.05) ROS generation compared to that in UVB-irradiated control HaCaT cells. Pretreatment with AE extract reversed reduction of Nrf-2 and SOD-1 protein expression and induction of HO-1 protein expression caused by UVB exposure in HaCaT cells, whereas it did not affect catalase expression. Conclusion: AE extract in 70% ethanol demonstrated a protective effect against UVB-induced oxidative stress and decreased expression of Nrf-2 and SOD-1 in human keratinocytes. These findings suggest that AE ethanol extract might have potential as a natural resource for a skin anti-photoaging product in the food and cosmetic industry.