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Anti-inflammatory Activity of Antimicrobial Peptide Papiliocin 3 Derived from the Swallowtail Butterfly, Papilio xuthus (호랑나비 유래 항균 펩타이드 파필리오신 3의 항염증 활성)

  • Shin, Yong Pyo;Lee, Joon Ha;Kim, In-Woo;Seo, Minchul;Kim, Mi-Ae;Lee, Hwa Jeong;Baek, Minhee;Kim, Seong Hyun;Hwang, Jae Sam
    • Journal of Life Science
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    • v.30 no.10
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    • pp.886-895
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    • 2020
  • The development of novel peptide antibiotics with potent antimicrobial activity and anti-inflammatory activity is urgently needed. In a previous work, we performed an in-silico analysis of the Papilio xuthus transcriptome to identify putative antimicrobial peptides and identified several candidates. In this study, we investigated the antibacterial and anti-inflammatory activities of papiliocin 3, which was selected bioinformatically based on its physicochemical properties against bacteria and mouse macrophage Raw264.7 cells. Papiliocin 3 showed antibacterial activities against E. coli and S. aureus without inducing hemolysis and decreased the nitric oxide production of the lipopolysaccharide-induced Raw264.7 cells. Moreover, ELISA and Western blot analysis revealed that papiliocin 3 reduced the expression levels of pro-inflammatory enzymes, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2). In addition, we examined whether papiliocin 3 could inhibit the expression of pro-inflammatory cytokines (interleukin-6 and interleukin-1β) in LPS-induced Raw264.7 cells. We found that papiliocin 3 markedly reduced the expression level of cytokines through the regulation of mitogen-activated protein kinases (MAPK) and nuclear factor kappa B (NF-κB) signaling. We also confirmed that papiliocin 3 binds to bacterial cell membranes via a specific interaction with lipopolysaccharides. Collectively, these findings suggest that papiliocin 3 could be a promising molecule for development as a novel peptide antibiotic.

Changes in the Profiles of Serum LH, Testosterone, Estrogen and IFG-I during Sexual Development in Male Korean Native Chickens (한국재래 수탉에서 부화 후 혈청내 LH, Testosterone, Estrogen과 IGF-I 농도의 변화)

  • Tae H. J.;Jang B. G.;Choi C. H.;Park Y. J.;Yang H. H.;Kim I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.2
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    • pp.135-141
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    • 2005
  • Changes in serum estradiol, insulin-like growth factor-1, leuteinizing and testosterone levels, and leuteinizing hormone-stimulated testosterone production per testis in vitro from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The changes in the profiles of the levels in the incubation medium of luteinizing hormone-stimulated (100 ng/mL) testosterone secretion per testis in vitro, and the serum LH, testosterone, estradiol, and insulin-like growth factor-I were determined by radioimmunoassay. Serum estradiol levels were not significantly different at week 4 compared to that of 8, 12, 16, 21, 32, :md 44. Significant decreases were observed at weeks 52 and 64. Serum leuteinizing hormone concentrations were not significantly different from 1 week to 12 weeks, increased gradually up to 32 weeks of age, and declined significantly thereafter; the highest value was at 32 weeks, and the lowest value was detected at 2 weeks of age. Serum insulin-like growth factor-I concentrations increased significantly from 1 week to 16 weeks, remained low and unchanged with advancing age. Serum testosterone concentrations were not significantly different at week 1 compared weeks 2, 4, 6, and 8. Significant increases were observed from 10 weeks to 32 weeks of age. Values at weeks 24, 28 and 32 and at weeks 32, 44, 52, and 64 were not significantly different. The highest value was at weeks 28 and the lowest value was detected at weeks 1 week. LH-stimulated testosterone production per testis in vitro increased gradually with age from 1 to 32 weeks and decreased significantly from 44 weeks to 64 weeks of age.

Comparative Study on Monetary Estimates of Natural Environment and Cultural Relics in Gyeongju National Park (경주국립공원의 문화유적과 자연환경의 가치추정 비교연구)

  • Kang, Kee-Rae;Kim, Dong-Pil;Baek, Jae-Bong
    • Korean Journal of Environment and Ecology
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    • v.26 no.2
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    • pp.273-282
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    • 2012
  • This study has estimated Gyeongju National Park's natural environment and cultural relic value in the same way and then been performed to compare the size of the value. Representative method to measure environmental property is contingent valuation methods, CVM. The variables and estimated models adopted for the calculation were same and the respondents were asked by distinguishing between the amount which they would pay to preserve the natural environment and that which they were willing to pay to preserve the cultural relics. As the result, WTP(Willing to pay), the amount that they were willing to pay to preserve the natural environment of Gyeongju National Park was 17,838 won per person and that to preserve the cultural relics appeared to be 316,248 won per person. Based on this, it was estimated that the value of the natural environment with which Gyeongju National Park provided annual visitors was 47 billion won and that the annual value of the cultural relics was 845.7 billion. If the natural environment and the cultural relics value elements are united, it can be estimated that the natural environment and cultural relic value got at the time of people's first visit to Gyeongju National Park is 334,086 won and that the annual value is 893.4 billion won. In this study, the value of the cultural relics has been estimated 18 times higher than that of the natural environment. This reason was that visitors judged that a total of 66 cultural properties including 11 national treasures, 23 treasures, 13 historic places, one historic sites and scenic spot and 18 local cultural properties, etc. which were distributed in Gyeongju National Park were worth far more than the natural environment. Based on the result of this study, the operating management plan of Gyeongju National Park should include a differentiated operation strategy through consultation with relevant experts by taking into account characteristics of the physical components.

Physiological Regulation of Luteinizing Hormone(LH) Expression in Rat Mammary Gland during Differentiation (분화중인 흰쥐 유선내 Luteinizing Hormone (LH) 유전자 발현의 생리적인 조절)

  • 이성호
    • Development and Reproduction
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    • v.5 no.2
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    • pp.175-180
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    • 2001
  • The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

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Effect of Di(2-ethyl hexyl)phthalate(DEHP) on the Onset of Puberty in Female Rat (암컷 흰쥐의 사춘기 개시에 미치는 di(2-ethyl hexyl)phthalate(DEHP)의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.2
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    • pp.147-154
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    • 2006
  • Phthalates such as di(2-ethyl hexyl)phthalate(DEHP) are industrial chemicals with wide-ranging human exposures because of their use in plastics and other common consumer products. Consequently, their adverse effects as endocrine disruptor in the reproductive physiology of both laboratory rodents and human have been studied extensively. The present study was undertaken to examine whether prepubertal exposure to DEHP affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. DEHP(100mg/kg/day) was administered daily from postnatal day 25(PND 25) through the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day. Gross anatomy and weight of reproductive tissues were compared to test the DEHP's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. Specific radioimmunoassay was carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed VO was shown in the DEHP group(PND $37.3{\pm}0.7$) compared to the control group(PND $35.3{\pm}0.7$; p<0.05). DEHP treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.05). Interestingly, elevation of serum LH levels was shown in the DEHP group(p<0.05). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals. Numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from DEHP-treated animals. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the DEHP-treated group. These effects were probably due to the inhibitory effects of DEHP on the synthesis and secretion of estrogen from granulosa cells. In the semiquantitative RT-PCR studies, the transcriptional activities of PR in both ovary(p<0.05) and uterus(p<0.01) from DEHP-treated animals were significantly lower than those from the control animals. The present studies demonstrated that the acute exposure to DEHP during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.

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Report on the Stability of Freeze-dried Standard Solution (동결건조표준액의 안정성에 관한 보고)

  • Park, Jun Mo;Yoo, Hye Jung;Kim, Han Chul;Han, Geul Soon
    • The Korean Journal of Nuclear Medicine Technology
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    • v.16 no.2
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    • pp.139-148
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    • 2012
  • Purpose : Since standard solution is the one that knows its exact concentration, the curve of the dissolution has been determined according to the amount of the solution, compared to the amount of the unknown sample. Therefore, the antigen that makes up standard materials should be made in a pure form. The configuration of the standard substance solution in the kit we use is a freeze-dried material, or made and comes as a liquid. Lyophilized reference material is used after dissolving in usually D.W. (Distilled Water), and if the antigen to use is too sensitive, reagents should be freeze-dried. Furthermore, when freeze-dried reference has to be frozen again after being dissolved, it should be kept under $-20^{\circ}C$ until the expiration date according to the reports. Since it is not expressed in the experiment if it is safe or stable to reuse the solution which was dissolved a few times, thus, this time it is tested and evaluated that the changes of the standard solution by freezing and melting several times, and its results and the effectiveness of it were compared to the solution which was kept in a fridge. Materials and Methods : Among Vitro diagnostic kits on the market made by radioimmunoassay, parathyroid hormone (PTH), adrenocorticotropic hormone (ACTH), luteinizing hormone (LH) are made of freeze-dried standard solution and all composed of the same Lot.NO. These hormones melted in D.W. and were separated into three groups. In the first group, melting and freezing were repeated, and in the second group, The solution only for one time use was put into a test tube after melting and freeze it. The third group was kept in the refrigerator. This experiment has been conducted from January to February in 2012. January to 2012. PH test was employed because ph is prone to changing depending on the change of protein. Each group of the standard solution, cpm (counter per minute), and the patient relative concentration values were compared by date, and Through the correlation coefficient and Paired t-test, the significant level of each group was analyzed. Results : ACTH, PTH, LH pH values were too subtle denaturation rather than numerical changes in the protein. In addition, when the standard solution of ACTH, PTH, LH was refrigerated, after 3 days and 7 days, there was a significant difference observed between the solution being kept in a refrigerator and a freezer within a significance level. Conclusion : Standard solution should be kept in a freezer, and being kept in a fridge, it is recommended to use the solution as soon as possible.

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Preliminary Study for Elevated Serum CXCL10 and CXCL11 in Active Pulmonary Tuberculosis Compared with the Other Pulmonary Diseases (타 폐질환과 비교를 통한 활동성 결핵에서 혈중 CXCL10과 CXCL11 증가의 의의)

  • Park, Mi Young;Kim, Shine Young;Hwang, Sang-Hyun;Kim, Ji-Eun;Lee, Min Ki;Lee, Chang-Hun;Lee, Eun-Yup
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.3
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    • pp.205-210
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    • 2009
  • Background: CXCL10 and CXCL11, which are family of CXCR3 ligands, are expressed by lymphocytes and even by bronchial epithelial cells if the cellular immunity is activated. This study evaluated the potential utility of CXCL10 and CXCL11 in the serum for active pulmonary tuberculosis in comparison with lung cancer, which activates the cellular immunity, and benign lung diseases. Methods: Patients who newly visited Pusan National University Hospital from January 2007 to December 2007 and were suspected of having lung cancer or tuberculosis were enrolled prospectively. The patients were classified pathologically and clinically into three groups, 47 with lung cancer, 18 with active pulmonary tuberculosis and 38 control patients with benign pulmonary disease. ELISA was used to determine the levels of CXCL10 and CXCL11 were determined in the serum. Results: The level of CXCL10 and CXCL11 were significantly higher in the active pulmonary tuberculosis group than in the lung cancer and benign lung disease groups (p<0.001, Kruskal-Wallis). The level of CXCL11 was significantly higher in the lung cancer group than in the benign pulmonary disease group, but there was no significant difference in level of CXCL10 between the three groups (p<0.001, p=0.655, respectively, Mann-Whitney U). The level of CXCL10 in patients with stage III+IV lung cancer was significantly higher than those with stage I+II, but there was no significant difference in the level of CXCL11 between the groups (p<0.001, p=0.07, respectively, Mann-Whitney U). There was no significant difference in the level of CXCL10 and CXCL11 between those with the presence and absence of lung cancer metastasis. There was a significant correlation between the level of CXCL10 and CXCL11 (r=0.223, p<0.001). Conclusion: CXCL10 and CXCL11 may be a potential useful markers for active pulmonary tuberculosis if used alongside other diagnostic methods.

Comparison of the Serum Cholesterol, Insulin Resistance and Markers of Metabolic Syndrome Based on Hepatitis C Virus RNA (C형 간염 바이러스 RNA 유무에 따른 지질, 인슐린저항성 및 대사증후군 지표 수준의 차이)

  • Cho, Sung-Hwan;Kim, Yun-Jin;Lee, Sang-Yeoup;Cho, Byung-Mann;Hwang, Hye-Lim;Yi, Yu-Hyeon;Cho, Young-Hye;Tak, Young-Jin;Jeong, Dong-Wook;Lee, Seung-Hun;Lee, Jeong-Gyu
    • Journal of agricultural medicine and community health
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    • v.41 no.4
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    • pp.205-216
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    • 2016
  • Objectives: We compared the difference of lipid, insulin resistance and metabolic markers based on HCV RNA in Korean adults.Methods: This was a cross-sectional study of 222 subjects visited the health promotion center of Pusan nationaluniversity hospital from 2004 to 2007. Subjects were anti-HCV antibody positive and were performed RT-PCR for HCV RNA. The HCV RNA (+) group were 85 subjects, HCV RNA (-) control group were 115 subjects, and the HCV RNA (-) but past positive group were 22 subjects. We performed anthropometry, anti-HCV, RT-PCR, plasma concentrations of insulin, total cholesterol, LDL-cholesterol, HDL-cholesterol, and triglyceride.Results: BMI, waist circumference, blood pressure, fasting plasma glucose, triglyceride, HDL cholesterol, insulin resistance such as HOMA-IR and QUICKI were not significantly different between HCV RNA positive and negative groups. The serum total cholesterol and LDL cholesterol level were significantly lower in the HCV RNA positive group than in the negative group ($186.24{\pm}37.63$ vs $197.22{\pm}37.23$ mg/dl, p=0.041, $111.66{\pm}34.06$ vs $121.38{\pm}35.50$ mg/dl, p=0.042). After adjusting age and sex, high total cholesterol (${\geq}200mg/dl$) (adjusted OR=0.51, 95%CI 0.28-0.94, p=0.03) and high LDL cholesterol (${\geq}130mg/dl$) (adjusted OR=0.46, 95%CI 0.24~0.87, p=0.02) were inversely associated with being HCV RNA positive (p<0.05). Conclusion: The serum total cholesterol and LDL-cholesterol level were significantly lower in HCV RNA (+) group than in HCV RNA (-) group, but not in HCV RNA (-) but past positive group. Prospective cohort studies are needed to clarify the relationship between HCV RNA and metabolic markers.

Preparation of $^{125}I-Iodotyraminehemisuccinyltaxol\;(^{125}ITHT)$ for Competitive Taxol Radioimmunoassay (Taxol 방사면역측정을 위한 $^{125}I-Iodotyraminehemisuccinyltaxol\;(^{125}ITHT)$의 제조)

  • Lee, Tae-Sup;Awh, Ok-Doo;Choi, Tae-Hyun;Kim, Hyun-Suk;Hong, Jun-Pyo;Lee, Eun-Sook;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.2
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    • pp.121-132
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    • 2002
  • Purpose : Taxol(Paclitaxel), an antineoplastic agent, has been used in the treatment of ovarian and breast cancers. The determination of optimal Taxol concentrations in human serum was required for enhancing therapeutic effect and maintaining the appropriate Taxol level in blood. This study was aimed to synthesizeradiolabeled Taxol derivatives as radiotracer in competitive radioimmunoassay for monitoring Taxol concentrations in blood and to determine the usefulness of its derivatives. Materials and Methods : Hemisucdcinyltaxol(HT) was synthesized by esterification of Taxol with succinic anhydride. Tyraminehemisuccinyltaxol(THT) was synthesized by coupling of HT with tyramine using isobutylchlormate as coupling agent and purified by HPLC. By using chloramine-T($5.25mg/ml,\;10{\mu}{\ell}$) as oxidant agent, THT($4mg/ml,\;30{\mu}{\ell}$) was labeled wity $^{125}I\;(37MBq,\;1mCi)$. To estimate the stability of purified THT, $^{125}I-iodotyraminehemisuccinyltaxol(^125}ITHT)$ was dissolved in 80% acetonitrile aqueous solution, and the solution was incubated at $4^{\circ}C\;and\;37^{\circ}C$ for 7 days. At various time intervals, the stability of THT and $^{125}ITHT$ was monitored. The titer of Taxol monoclonal antibody, 3G5A7, was determined by competitive radioimmunoassay using $^{125}ITHT$ as a labeled antigen. A standard dose-response curve was demonstated by Taxol competitive radioimmunoassay. Resulls : HT and THT were synthesized with 79.9% and 19.5% yield, respectively. The labeling yield of $^{125}ITHT$ was 93%. After 7 days, the chemical purity of THT was 96.5% at $4^{\circ}C$, and 97.5% at $37^{\circ}C$. After 3 days, $^{125}ITHT$ was stable with 94.7% at $4^{\circ}C$ and 93.4% at $37^{\circ}C$. After 7 days, fadiochemical purity was diminished to 88.1% at $37^{\circ}C$. The titer of Taxol monoclonal antibody, 3G5A7, was determined to 1:256. A standard dose-response curve demonstated good collinearity ($R^2=0.971$) as Taxol concentration-dependent manner. Conclusion : Competitive radioimmunoassay using $^{125}I-iodotyraminehemisuccinytaxol$ as radiotracer could be used to monitor for concentration of Taxol in the human serum.

Study on Public Awareness of establishing Marine Protected Areas - Case Study of Guimaras Province, Philippines using Contingent Valuation Method (해양보호구역의 설정에 대한 대중인식도 연구 - 조건부 가치측정법을 이용한 필리핀 기마라스주의 사례)

  • Kim, Tae-Goun
    • Journal of Navigation and Port Research
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    • v.38 no.6
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    • pp.663-672
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    • 2014
  • In Korea and the Philippines, as well as all over the world, with the recognition of the importance of marine ecological resources, the marine protected areas(MPA) have been established and managed to protect and preserve these resources. While the number of marine protected areas for marine ecological resources protection has been increased, there is main problem that the most of MPAs do not achieve their intended management objectives. the effective management. Because of the positive and negative impacts on local communities and fishermen as direct stockholders, there has been ongoing debate on the pros and cons of implementing MPAs. Accordingly, this research conducted a case study of establishing Marine Protected Areas in Guimaras, Philippines because Philippines fisheries code of 1998 (Republic Act 8550), which is enacted to manage, conserve and protect fishery resources, obliged local governments to designate no less than 15% of jurisdictional municipal water as fisheries resource protection areas for a long time. To do this, a dichotomous-choice contingent-valuation survey was conducted in the two municipalities of Guimaras, Philippines to investigate public opinion in debates over MPAs and to estimate willingness to pay (WTP) for MPAs to protect and conserve marine habitats for fishery resources. Because of the expected economic costs by prohibiting fishing activities within the establishing newMPA, 58.7% of respondents thought the costs should be compensated, but 91.4% respondents voted in favor of increasing MPAs for fisheries resources as a protective measure. Finally, with Contingent Valuation Method(CVM), the aggregate mean WTP (375.5ha) of San Lorenzo and Sibunag residents in Guimaras Province, Philippines for establishing the additional MPA in their municipality waters was estimated to $1,046,791. Therefore, these findings could be used as a valuable data for establishing effective management plan of MPAs in Korea.