Melatonin Rescues Mesenchymal Stem Cells from Senescence Induced by the Uremic Toxin p-Cresol via Inhibiting mTOR-Dependent Autophagy

윤승필,Yong-seok Han,이준희,김상민,이상훈 한국응용약물학회 2018 Biomolecules & Therapeutics(구 응용약물학회지) Vol.26 No.4

p-Cresol, found at high concentrations in the serum of chronic kidney failure patients, is known to cause cell senescence and other complications in different parts of the body. p-Cresol is thought to mediate cytotoxic effects through the induction of autophagy response. However, toxic effects of p-cresol on mesenchymal stem cells have not been elucidated. Thus, we aimed to investigate whether p-cresol induces senescence of mesenchymal stem cells, and whether melatonin can ameliorate abnormal autophagy response caused by p-cresol. We found that p-cresol concentration-dependently reduced proliferation of mesenchymal stem cells. Pretreatment with melatonin prevented pro-senescence effects of p-cresol on mesenchymal stem cells. We found that by inducing phosphorylation of Akt and activating the Akt signaling pathway, melatonin enhanced catalase activity and thereby inhibited the accumulation of reactive oxygen species induced by p-cresol in mesenchymal stem cells, ultimately preventing abnormal activation of autophagy. Furthermore, preincubation with melatonin counteracted other pro-senescence changes caused by p-cresol, such as the increase in total 5′-AMP-activated protein kinase expression and decrease in the level of phosphorylated mechanistic target of rapamycin. Ultimately, we discovered that melatonin restored the expression of senescence marker protein 30, which is normally suppressed because of the induction of the autophagy pathway in chronic kidney failure patients by p-cresol. Our findings suggest that stem cell senescence in patients with chronic kidney failure could be potentially rescued by the administration of melatonin, which grants this hormone a novel therapeutic role.

세계보건기구(WHO)의 대유행 인플루엔자 대비체계(PIPF)와 나고야의정서의 관계

유예리,윤승필 강원대학교 비교법학연구소 2020 환경법과 정책 Vol.25 No.-

코로나19와 같은 공중보건 위기상황에서는 병원체 표본 및 그 유전자염기서열정보에 대한 제공이 신속히 이루어져야 한다. 그런데 국가가 자국의 국경 내에 있는 병원체 자원도 국가의 주권적 자산이라고 주장하면서 생물다양성협약과 그 부속조약인 나고야의정서에 따라 접근 및 이익 공유를 요구한다면 추가적 거래비용이 발생할 뿐만 아니라 백신과 치료제 개발도 지연될 수 있다. 물론 나고야의정서 제8(b)조는 인간, 동물 또는 식물 건강에 위해를 줄 수 있는 긴급 상황이 발생할 경우 상당한 주의를 기울여야 하고, 관련 유전자원의 접근 및 이익 공유 절차의 간소화를 고려하도록 규정하고 있다. 즉 동 조항은 당사국들에게 신종 인플루엔자 등과 같은 전염병이 발생할 경우, 백신 및 치료제 개발 등에 필요한 병원체 등 유전자원에 대한 접근 절차를 간소화할 수 있는 재량권을 부여하고 있다. 한편 세계보건기구는 보건의료 위기상황에 대비하여 2011년 대유행 인플루엔자 대비체계(PIPF)를 채택하였다. 2006년 12월 인도네시아 정부가 생물다양성협약을 근거로 자국의 국경 내에 있는 바이러스에 대한 주권을 주장하며 세계보건기구와의 바이러스 표본 공유를 거절하자 세계보건기구가 채택한 결의안이다. 즉 PIPF는 세계보건기구 회원국들이 대유행 잠재력을 지닌 인간 인플루엔자 바이러스에 대한 접근과 진단, 백신, 항바이러스 등 사용과 관련된 이익 공유를 규율하는 다자간 문서이다. 이처럼 병원체의 접근 및 이익 공유라는 동일한 사안을 두고 이를 규율하는 국제협약과 국제문서가 중복적으로 존재하는 상황에서 양자의 모호한 관계는 향후 전염병 대유행이 재발할 경우 국제적으로 신속한 협력과 대응을 가로막고, 과학기술의 발전도 저해할 수 있다. 즉 양 문서간의 관계를 명확히 정립하지 않으면, 급박한 보건의료 위기상황에서 나고야의정서의 이행으로 인한 세계보건기구의 대응과 일치하지 않는 상황이 발생할 수 있다. 따라서 나고야의정서와 세계보건기구가 채택한 문서인 PIPF 간의 관계를 정립할 필요가 있다. 이에 본고는 먼저 나고야의정서가 이행되고 있는 상황에서 세계보건기구가 병원체 대상 별도의 접근 및 이익 공유 체계를 마련할 수 있는지 양자 간의 관계부터 살펴보고자 한다. 나고야의정서 제3조는 동 의정서의 적용범위에 병원체를 제외한다고 명시하지 않고 있고, PIPF이 나고야의정서 제4.4조상의 특별국제문서에 해당하는지를 두고 해석의 여지가 있다. 다음으로 PIPF의 적용대상 및 범위를 살펴보고, 나고야의정서가 적용될 수 없는 요건, 다시 말해서 PIPF가 적용되는 요건들을 알아보고자 한다. In public health crises such as COVID-19, pathogen samples and gene sequence data should be publically available to international organizations in a timely manner. However, if the federal government claims the intellectual ownership of the pathogen resources, and requests access and benefit sharing in accordance with the Convention on Biological Diversity and the Nagoya Protocol, the additional transaction costs will delay the development of vaccine and treatments. Article 8(b) of the Nagoya Protocol states that due regard should be paid in the event of an emergency that threaten or damage human, animal or plant health, and that expeditious access to genetic resources and fair sharing of benefits arising out of the use of such genetic resources should be considered. In other words, in the event of an outbreak of infectious diseases such as H5N1, the parties shall consider expeditious access procedures to genetic resources such as pathogens needed for the development of vaccines and treatments. Meanwhile, the World Health Organization(WHO) adopted the Pandemic Influenza Preparedness(PIPF) in 2011 to prepare for health and medical crises. This resolution was adopted by the World Health Organization in December 2006 after the Indonesian government refused to share virus samples with the World Health Organization, claiming sovereignty over viruses within its borders based on the Convention on Biological Diversity. PIPF is a multilateral instrument that regulates access to human influenza viruses with pandemic potential and sharing of benefits related to their use, including diagnosis, vaccines and antiviral agents. In this situation where there are separate international agreements and instruments that discipline the issue of access to and sharing benefits of pathogens, the ambiguous relationship between the two could hinder international cooperation and response to future pandemics, and also prevent the development of science and technology. Namely, if the relationship between the two instruments is not established, the implementation of Nagoya Protocol in an urgent health and medical crisis may hinder and delay the WHO’s response to the sharing of pathogens. Therefore, it is necessary to establish a relationship between the PIPF, an instrument adopted by the World Health Organization, and the Nagoya Protocol. Article 3 of the Nagoya Protocol does not specify that pathogens are excluded from the scope of the protocol, and Article 4.4 of the Protocol does not specify whether the PIPF falls under specialized international instruments, leaving room for subjective interpretation. In response, this paper will first look at the relationship between the two to see if WHO can establish a separate ABS system for pathogens in the context of the implementation of the Protocol. Next, it will examine the application and scope of the PIPF and identify the circumstances in which the protocol cannot be applied. In other words, the requirements to which PIPF is applied. Therefore, in this review, we would like to examine and suggest the legislative and policy tasks that Korea should implement internally and externally.

합성생물학의 발전과 EU의 법적 대응방안에 대한 연구 - 유럽법원(EuGH)의 C-528/16 판결에 대한 평석을 중심으로 -

박신욱 ( Park Shin-uk ),윤승필 ( Yun Seung-pil ) 경상대학교 법학연구소 2020 法學硏究 Vol.28 No.2

학문ㆍ연구의 자유에 대한 개입을 정당화하거나 이와 관련된 법익을 형량함에 있어 정당성을 확보하기 위한 일반적인 규정은 존재할 수 없다. 연구의 위험성이 크고 구체적일 수록 학문ㆍ연구의 자유에 대한 개입의 정당화가 이루어질 수 있다는 정도의 원칙만이 존재할 뿐이다. 그렇기 때문에 위험성이 큰 연구의 경우 국가는 사전적 조치를 취할 수도 있게 된다. 이와 관련하여 본고(本稿)에서는 합성생물학의 발전에 따른 EU의 사전적 조치에 대해 유럽법원의 C-528/16 판결에 대한 평석을 통해 평가하고자 하였다. 유럽법원은 프랑스 국사원(Conseil d’État)의 청문에 따라 돌연변이 유발기술의 발전에 따른 법적 쟁점에 대해 판단하였다. 특히 돌연변이 유발로 인해 생성된 유기체가 유전자 변형생물체에 해당하는지 여부(i), 새로운 돌연변이 유발방식의 경우에는 기존의 돌연변이 유발방식과 마찬가지로 2001년 지침의 적용이 배제되는지 여부(ii)에 대해 판단하게 된 것이다. 유럽법원은 첫 번째 의문사항과 관련하여 유전자변형생물체에 해당한다고 판단하였고, 두 번째 의문사항과 관련해서는 2001년 지침의 적용이 배제되지 않는다고 판단하였다. 생각건대, 이러한 유럽법원의 판단은 자연과학에 대한 이해의 한계를 명확하게 보여준다. 특히 유럽법원이 “효과”라는 측면을 강조함으로써 새로운 돌연변이 유발기술을 이식 유전자 기술과 동일하게 판단한 점은 비판을 피하기 어려워 보인다. 또한 유럽법원의 판결은 법률과 기술의 발전으로 인한 현실의 간극을 명백하게 보여주고 있으며, 현재의 과학기술의 안정성과도 모순되는 결과를 이끌어냈다. 이와 달리 법률고문 Bobek의 의견은 향후 우리가 나아가야 할 방향성에 대해 명백히 보여주고 있다고 판단된다. Bobek은 기술 및 학문의 진보에 초점을 맞추고 사전예방원칙의 남용을 경계하였다는 점에서 더 높은 평가가 이루어져야 할 것이다. Es gibt keine allgemeinen Bestimmungen, um das Eingreifen in die Freiheit von Wissenschaft und Forschung zu rechtfertigen. Es gibt nur einen Grundsatz: Je größer und spezifischer das Forschungsrisiko ist, desto gerechtfertigter ist der Eingriff in die Freiheit von Wissenschaft und Forschung. Infolgedessen kann ein Staat mit hohen Risiken möglicherweise vorsorgliche und vorläufige Massnahmen ergreifen. In diesem Zusammenhang wurde in dieser Untersuchung beabsichtigt, die vorläufigen Maßnahmen der EU zur Entwicklung der synthetischen Biologie durch die Überprüfung der Entscheidung des Europäischen Gerichtshofs (EuGH, 25.07.2018 - C-528/16) zu bewerten. In der Vorlage des französischen Conseil d'État hat der Europäische Gerichtshof über rechtliche Fragen entscheidet, die sich aus der Entwicklung und dem Fortschritt der Mutagenesetechnologie ergeben. Insbesondere hat der Europäische Gerichtshof festgestellt, ob durch Mutagenese erzeugte Organismen gentechnisch veränderte Organismen sind (i) und ob bei neuer Mutagenese die Richtlinie von 2001 wie bei herkömmlichen Mutagenesemethoden ausgeschlossen ist (ii). Der Europäische Gerichtshof war der Auffassung, dass es sich in Bezug auf die erste Frage um einen genetisch veränderten Organismus handelt und dass die Anwendung der Leitlinien von 2001 in Bezug auf die zweite Frage nicht ausgeschlossen wurde. Meiner Meinung nach zeigt dieses Urteil des Europäischen Gerichtshofs deutlich die Grenzen unseres Verständnisses der Naturwissenschaften auf. Insbesondere scheint es schwierig zu sein, die Kritik zu vermeiden, dass der Europäische Gerichtshof die neue Mutagenesetechnologie genauso schätzt wie die Transgen-Technologie, indem er den Aspekt „Wirkung“ hervorhebt. Darüber hinaus zeigt das Urteil des Europäischen Gerichtshofs deutlich die Kluft zwischen der Realität und Recht aufgrund der Entwicklung von Technologie und führt zu einem Widerspruch zur Stabilität der gegenwärtigen Wissenschaft und Technologie. Im Gegensatz dazu zeigt Bobeks Meinung deutlich, in welche Richtung wir in Zukunft gehen sollten. Bobek sollte einen hohen Stellenwert haben, sich auf technologische und akademische Fortschritte konzentrieren und sich vor dem Missbrauch von Vorsorgeprinzipien schützen.

Multidetector computed tomographic angiography evaluation of micropig major systemic vessels for xenotransplantation

유정민,윤웅,박재홍,윤승필,장민우,한호재 대한수의학회 2011 Journal of Veterinary Science Vol.12 No.3

Due primarily to the increasing shortage of allogeneic donor organs, xenotransplantation has become the focus of a growing field of research. Currently, micropigs are the most suitable donor animal for humans. However, no standard method has been developed to evaluate the systemic vascular anatomy of micropigs and standard reference values to aid in the selection of normal healthy animals as potential organ donors are lacking. Using 64-channel multidetector row computed tomographic angiography (MDCTA), we evaluated morphological features of the major systemic vessels in micropigs and compared our results to published human data. The main vasculature of the animals was similar to that of humans, except for the iliac arterial system. However, diameters of the major systemic vessels were significantly different between micropigs and humans. Specifically, the diameter of the aortic arch, abdominal aorta, external iliac artery, and femoral artery, were measured as 1.50 ± 0.07 cm, 0.85 ± 0.06 cm, 0.52 ± 0.05 cm, and 0.48 ± 0.05 cm, respectively, in the micropigs. This MDCTA data for micropig major systemic vessels can be used as standard reference values for xenotransplantation studies. The use of 64-channel MDCTA enables accurate evaluation of the major systemic vasculature in micropigs.

Cell-Based Screen Using Amyloid Mimic β23 Expression Identifies Peucedanocoumarin III as a Novel Inhibitor of α-Synuclein and Huntingtin Aggregates

함상우,김효정,황서진,강현욱,윤승필,김상준,김동훈,권현숙,이윤송,조명래,신흥묵,최희정,정가영,고한석,이금화,이연종 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.6

Aggregates of disease-causing proteins dysregulate cellular functions, thereby causing neuronal cell loss in diverse neurodegenerative diseases. Although many in vitro or in vivo studies of protein aggregate inhibitors have been performed, a therapeutic strategy to control aggregate toxicity has not been earnestly pursued, partly due to the limitations of available aggregate models. In this study, we established a tetracycline (Tet)-inducible nuclear aggregate (β23) expression model to screen potential lead compounds inhibiting β23-induced toxicity. High-throughput screening identified several natural compounds as nuclear β23 inhibitors, including peucedanocoumarin III (PCIII). Interestingly, PCIII accelerates disaggregation and proteasomal clearance of both nuclear and cytosolic β23 aggregates and protects SH-SY5Y cells from toxicity induced by β23 expression. Of translational relevance, PCIII disassembled fibrils and enhanced clearance of cytosolic and nuclear protein aggregates in cellular models of huntingtin and α-synuclein aggregation. Moreover, cellular toxicity was diminished with PCIII treatment for polyglutamine (PolyQ)-huntingtin expression and α-synuclein expression in conjunction with 6-hydroxydopamine (6-OHDA) treatment. Importantly, PCIII not only inhibited α-synuclein aggregation but also disaggregated preformed α-synuclein fibrils in vitro. Taken together, our results suggest that a Tet-Off β23 cell model could serve as a robust platform for screening effective lead compounds inhibiting nuclear or cytosolic protein aggregates. Brain-permeable PCIII or its derivatives could be beneficial for eliminating established protein aggregates.

Comparison of cardiac function and coronary angiography between conventional pigs and micropigs as measured by multidetector row computed tomography

안영근,유정민,정해창,김윤현,정명호,이민영,Sang Hun Lee,Jae Hong Park,윤승필,한호재 대한수의학회 2008 Journal of Veterinary Science Vol.9 No.2

Pigs are the most likely source animals for cardiac xenotransplantation. However, an appropriate method for estimating the cardiac function of micropigs had not been established. Computed tomography (CT) analysis aimed at estimating cardiac function and assessing the coronary arteries has not been carried out in micropigs. This study determined the feasibility of evaluating cardiac function in a micropig model using multidetector row computed tomography (MDCT) and compared the cardiac function values with those of conventional pigs. The mean age of the conventional pigs and micropigs was approximately 80 days and approximately 360 days, respectively. The mean body weight in the conventional pigs and micropigs was 29.70 ± 0.73 and 34.10 ± 0.98 kg, respectively. Cardiac MDCT detected ejection fractions of 52.93 ± 3.10% and 59.00 ± 5.56% and cardiac outputs of 1.46 ± 0.64 l/min and 1.21 ± 0.24 l/min in conventional pigs and micropigs, respectively. There were no significant differences in cardiac function between conventional pigs and micropigs in the reconstructed CT images. There were also no differences in the coronary angiographic images obtained by MDCT. It is expected that the results of this study will help improve understanding of cardiac function in micropigs. The data presented in this study suggest that MDCT is a feasible method for evaluating cardiac function in micropigs.

64-Channel multi-detector row CT angiographic evaluation of the micropigs for potential living donor lung transplantation

윤웅,유정민,이민영,Yong Ju Moon,Sang Hun Lee,박재홍,윤승필,Min Woo Jang,박성수,한호재 대한수의학회 2010 Journal of Veterinary Science Vol.11 No.3

Micropigs are the most likely source animals for xenotransplantation. However, an appropriate method for evaluating the lung of micropigs had not been established. Therefore, this study was performed to evaluate the feasibility of 64-channel multi-detector row computed tomography (MDCT) to measure the diameter of the pulmonary arteries and the lung volume in micropigs. The mean diameters of the trachea, and left and right bronchi were 1.6 ± 0.17, 1.18 ± 0.14,and 1.1 ± 0.11 cm, respectively. The mean diameters of the main, right, and left pulmonary arteries were 1.38 ± 0.09, 1.07± 0.26, and 0.98 ± 0.13 cm and the diameters of right, left, and common inferior pulmonary veins were 0.97 ± 0.20, 0.76 ±0.20, and 1.99 ± 0.26 cm, respectively. The mean lung volume was 820.3 ± 77.11 mL. The data presented in this study suggest that the MDCT may be a noninvasive, rapid, and accurate investigational method for pulmonary anatomy in living lung donors.

Imaging evaluation of the liver using multi-detector row computed tomography in micropigs as potential living liver donors

유정민,김동현,Min Young Lee,Sang Hun Lee,박재홍,윤승필,Min Woo Jang,김성환,노규진,한호재 대한수의학회 2009 JOURNAL OF VETERINARY SCIENCE Vol.10 No.2

The shortage of organ donors has stimulated interest in the possibility of using animal organs for transplantation into humans. In addition, pigs are now considered to be the most likely source animals for human xenotransplantation because of their advantages over non-human primates. However, the appropriate standard values for estimations of the liver of micropigs have not been established. The determination of standard values for the micropig liver using multi-detector row computed tomography (MDCT) would help to select a suitable donor for an individual patient, determine the condition of the liver of the micropigs and help predict patient prognosis. Therefore, we determined the standard values for the livers of micropigs using MDCT. The liver parenchyma showed homogenous enhancement, and had no space-occupying lesions. The total and right lobe volumes of the liver were 698.57 ± 47.81 ml and 420.14 ± 26.70 ml, which are 51.74% and 49.35% of the human liver volume, respectively. In micropigs, the percentage of liver volume to body weight was approximately 2.05%. The diameters of the common hepatic artery and proper hepatic artery were 6.24 ± 0.20 mm and 4.68 ± 0.13 mm, respectively. The hepatic vascular system of the micropigs was similar to that of humans, except for the variation in the length of the proper hepatic artery. In addition, the diameter of the portal vein was 11.27 ± 0.38 mm. In conclusion, imaging evaluation using the MDCT was a reliable method for liver evaluation and its vascular anatomy for xenotransplantation using micropigs.

조직재생치료에서 줄기세포의 활용

허정선,김윤희,이민영,이상훈,박재홍,김미옥,나순임,김민희,서한나,이유진,류정민,윤승필,한호재 全南大學校醫科學硏究所 2007 전남의대학술지 Vol.43 No.1

Multidetector row computed tomography evaluation of the micropig kidney as a potential renal donor

윤웅,이민영,유정민,Yong Ju Moon,Sang Hun Lee,박재홍,윤승필,Min Woo Jang,박성수,한호재 대한수의학회 2010 JOURNAL OF VETERINARY SCIENCE Vol.11 No.1

Multidetector row computed tomography (MDCT) provides anatomical information about the kidney and other internal organs. Presently, the suitability of 64-channel MDCT to assess the kidney of healthy micropigs was evaluated. Morphological evaluations of the kidney and the major renal vessels of six healthy micropigs were carried out using MDCT, recording kidney volume and the diameter and length of renal arteries and veins. The mean diameters and lengths of the renal artery were 0.44 ± 0.05 and 4.51 ± 0.55 cm on the right side and 0.46 ± 0.06 and 3.36 ± 0.27 cm on the left side, respectively. The mean diameters and lengths of the renal vein were 1.44 ± 0.52 and 4.22 ± 1.29 cm on the right side and 1.38 ± 0.17 and 5.15 ± 0.87 cm on the left side, respectively. The mean volume of the right kidney was 79.3± 14.5 mL and of the left kidney was 78.0 ± 13.9 mL. The data presented in this study suggest that the MDCT offers a noninvasive, rapid, and accurate method for the evaluation of the renal anatomy in living kidney donors. It also provides sufficient information about extra-renal anatomy important for donor surgery and determination of organ suitability.