Expression of the Blue Fluorescent Protein (AmCyan) in the Fibroin of Transgenic Silkworms

Seon Young Kim,Seong Wan Kim,Eun Young Yun,Kwang-Ho Choi,Seong Ryul Kim,Seok Woo Kang,Seung Won Park,Tae Won Goo 한국응용곤충학회 2014 한국응용곤충학회 학술대회논문집 Vol.2014 No.04

To product the blue fluorescent protein (AmCyan) expressed cocoon, we were fused AmCyan cDNA to the heavy chain gene and injected the gene into a silkworm. AmCyan was one of the existing violet fluorochromes and originally derived from the fluorescent protein amFP486. AmFP486 was cloned from the sea anemone Anemonia majano (GenBank accession number AF168421), and belongs to the family of fluorescent proteins (FPs) isolated from coral reef organisms. The AmCyan fusion protein, each with N- and C- terminal sequences or the fibroin H-chain, were designed to be secreted into the lumen of the posterior silk glands. The expression of the AmCyan/H-chain fusion gene was regulated by the fibroin H-chain promoter. The use of the 3xP3 EGFP as a marker allowed us to rapidly distinguish transgenic silkworm. Mixtures of the donor and helper vectors were micro-injected into 300 eggs of bivoltine silkworms (Baegokjam). EGFP fluorescence was observed in 3 broods of transgenic silkworms under a florescence stereomicroscope. The cocoon was displayed strong blue fluorescence, proving that the fusion protein was present in the cocoon. Accordingly, we suggest that the AmCyan gene expressed cocoon will be enable the production of the novel biomaterials based on the transgenic silk.

Hospital-based Influenza Morbidity and Mortality (HIMM) Surveillance for A/H7N9 Influenza Virus Infection in Returning Travelers

Song, Joon Young,Noh, Ji Yun,Lee, Jacob,Woo, Heung Jeong,Lee, Jin Soo,Wie, Seong-Heon,Kim, Young Keun,Jeong, Hye Won,Kim, Shin Woo,Lee, Sun Hee,Park, Kyung-Hwa,Kang, Seong Hui,Kee, Sae Yoon,Kim, Tae H KOREAN ACADEMY OF MEDICAL SCIENCE 2018 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.33 No.7

<P>Since 2013, the Hospital-based Influenza Morbidity and Mortality (HIMM) surveillance system began a H7N9 influenza surveillance scheme for returning travelers in addition to pre-existing emergency room (ER)-based influenza-like illness (ILI) surveillance and severe acute respiratory infection (SARI) surveillance. Although limited to eastern China, avian A/H7N9 influenza virus is considered to have the highest pandemic potential among currently circulating influenza viruses. During the study period between October 1st, 2013 and April 30th, 2016, 11 cases presented with ILI within seven days of travel return. These patients visited China, Hong Kong, or neighboring Southeast Asian countries, but none of them visited a livestock market. Seasonal influenza virus (54.5%, 6 among 11) was the most common cause of ILI among returning travelers, and avian A/H7N9 influenza virus was not detected during the study period.</P>

Distinct clinical courses according to presenting phenotypes and their correlations to <i>ATP7B</i> mutations in a large Wilson's disease cohort

Lee, Beom H.,Kim, Joo H.,Lee, Sun Y.,Jin, Hye Y.,Kim, Kwi‐,Joo,Lee, Jin‐,Joo,Park, Jung‐,Young,Kim, Gu‐,Hwan,Choi, Jin‐,Ho,Kim, Kyung M.,Yoo, Han‐,Wook Blackwell Publishing Ltd 2011 Liver International Vol.31 No.6

<P><B>Abstract</B></P><P><B>Introduction and aims: </B> Wide phenotypic and genotypic heterogeneities in Wilson's disease (WD) have been reported, hampering the study of their correlations. The goal of this study was to identify the factors related to these diversities.</P><P><B>Methods: </B> Clinical courses and molecular genetic characteristics were analysed in 237 unrelated Korean WD families. The average follow‐up period was 8.2 ± 5.8 years.</P><P><B>Results: </B> Presenting phenotypes were classified as H1 (12.2%), H2 (42.4%), N1 (21.6%), N2 (0.4%), NX (0.4%), presymptomatic (22.4%) and other (0.4%), modifying the guidelines by Ferenci and colleagues. Age at presentation was youngest and cirrhosis was rarest in the presymptomatic group. Decompensated cirrhosis was the highest in the H1 group. Favourable outcome was rarest in the N1 group. Forty‐seven (11 novel) <I>ATP7B</I> mutations were identified in 85% of the 474 alleles. Multiplex ligation‐dependent probe amplification assays in <I>ATP7B</I> and analyses of <I>ATOX1</I> and <I>COMMD1</I> genes identified no additional mutations. Yeast complementation assays demonstrated functional perturbation of the seven novel missense mutants. Five major mutations, p.Arg778Leu, p.Ala874Val, p.Asn1270Ser, p.Lys838SerfsX35 and p.Leu1083Phe, accounted for 63% of the alleles. H1 was more common, age at presentation was younger and N1+N2+NX tended to be less common in patients with nonsense, frame shifting or splicing mutations than in those with missense mutations alone. Patients with both mutations in the transduction (Td) or the ATP hinge domain showed presymptomatic or hepatic manifestations but no neurological manifestation.</P><P><B>Conclusions: </B> The presenting phenotype strongly affects the clinical outcome of WD, and is related to the <I>ATP7B</I> mutation type and location, providing an evidence for genotype–phenotype correlations in WD.</P>

한국에서 지역에 따른 Helicobacter pylori 균주 내성

김재연 ( Jae Yeon Kim ),김나영 ( Na Young Kim ),김성중 ( Sung Jung Kim ),백광호 ( Gwang Ho Baik ),김광하 ( Gwang Ha Kim ),김정목 ( Jung Mogg Kim ),남령희 ( Ryoung Hee Nam ),김홍빈 ( Hong Bin Kim ),이동호 ( Dong Ho Lee ),정현채 ( H 대한소화기학회 2011 대한소화기학회지 Vol.57 No.4

Background/Aims: This study was performed to compare the prevalence rates of primary antibiotic resistance in Helicobacter pylori (H. pylori) isolates among different regions of Korea. Methods: H. pylori were isolated from gastric mucosal biopsy specimens of 99 Koreans who lived in Gyeonggi (n=40), Kangwon province (n=40) and Busan (n=19) from April to August in 2008. All the patients had no history of H. pylori eradication therapy. The susceptibilities of the H. pylori isolates to amoxicillin, clarithromycin, metronidazole, tetracycline, azithromycin, ciprofloxacin, levofloxacin, and moxifloxacin were tested according to the agar dilution method. Results: There was a difference in resistance to clarithromycin in three institutes located among Gyeonggi (32.5%), Kangwon province (12.5%) and Busan (42.1%) by One way ANOVA test (p=0.027) and nonparametric Kruskal Wallis test (p=0.027). However, by post-hoc analysis, there was no statistically significant difference among three regions. Similarly, the other 7 antibiotics (amoxicillin, metronidazole, tetracycline, azithromycin, ciprofloxacin, levofloxacin and moxifloxacin) did not show any significant difference. Conclusions: There was no significant regional difference of the primary antibiotic resistance of H. pylori. However, the included patient number might not be enough for this conclusion demanding further evaluations. (Korean J Gastroenterol 2011;57:221-229)

Time-dependent effects of Klebsiella pneumoniae endotoxin on the pharmacokinetics of chlorzoxazone and its main metabolite, 6-hydroxychlorzoxazone, in rats: restoration of the parameters in 96 hour in KPLPS rats to control levels

Jung, Hye Y.,Kang, Hee E.,Choi, Young H.,Kim, So H.,Lee, Myung G. John Wiley Sons, Ltd. 2009 Biopharmaceutics and Drug Disposition Vol.30 No.8

<P>It has been reported that chlorzoxazone (CZX) was primarily metabolized via hepatic Cyp2e1 to form 6-hydroxychlorzoxazone (OH-CZX) in rats, and the activity of aniline hydroxylase (a Cyp2e1 marker) in the liver was significantly decreased in rats at 24 h after pretreatment with lipopolysaccharide derived from Klebsiella pneumoniae (24 h KPLPS rats), whereas the levels were not changed at 2 h and 96 h in the KPLPS rats. Thus, the time-dependent pharmacokinetic parameters of CZX and OH-CZX were evaluated after the intravenous administration of CZX (20 mg/kg) to control rats, and the 2 h, 24 h and 96 h KPLPS rats along with the time-dependent changes in the protein expression of hepatic Cyp2e1. After the intravenous administration of CZX to 24 h KPLPS rats, the AUC<SUB>0–2 h</SUB> of OH-CZX and AUC<SUB>OH-CZX, 0–2 h</SUB>/AUC<SUB>CZX</SUB> were significantly smaller (by 40.5% and 71.2%, respectively) than those of controls due to the significant decrease (by 75.3%) in the protein expression of hepatic Cyp2e1. However, in 96 h KPLPS rats, the pharmacokinetic parameters of both CZX and OH-CZX were unchanged compared with controls due to the restoration of the protein expression of hepatic Cyp2e1 to control levels. These observations highlighted the existence of the time-dependent effects of KPLPS on the pharmacokinetics of CZX and OH-CZX in rats. Copyright © 2009 John Wiley & Sons, Ltd.</P>

Farnesyl transferase 억제제인 YH3938 및 YH3945에 의한 Ras 발암원성 억제

Myung-Ju Oh(오명주),Nong Yeon Kim(김농연),Su-Eun Lim(임수은),Young-Hwa Chung(정영화),Byung H Jhun(전병학) 한국생명과학회 2010 생명과학회지 Vol.20 No.2

Ras 유전자는 30%의 인간암에서 변이가 발견되며 세 종류의 isoform, H-Ras, K-Ras 및 N-Ras로 구성되어 있다. Ras 단백질의 CAAX motif에 farnesylation과 같은 번역 후 변형은 Ras의 활성에 필수 요소이다. 본 연구에서는 새로운 farnesyl transferase 억제제인 YH3938과 YH3945의 발암원성 H-Ras, K-Ras 및 N-Ras의 작용에 대한 영향을 조사하였다. YH3938과 YH3945는 발암원성 H-Ras에 의해 형질전환된 Rat2 세포의 증식과 형태 변화를 억제하였으나 K-Ras에 대해서는 효과가 없었다. N-Ras에 대해서는 약한 영향이 있었다. H-Ras와 N-Ras에 의한 SRE promoter 활성화는 YH3938과 YH3945에 의해 억제되었으나, K-Ras에는 영향이 없었다. Ras 단백질의 bandshift 분석을 통해 YH3938은 H-Ras와 N-Ras의 번역 후 변환을 억제하였으나, K-Ras에는 영향이 없었다. YH3945는 H-Ras의 변환에만 영향이 있었다. 결론적으로 YH3938과 YH3945는 H-Ras의 farnesylation을 억제하여 그 발암원성을 억제하며, YH3938은 N-Ras 작용을 농도의존적으로 억제하며, K-ras에 대해서는 영향이 없음을 알 수 있었다. Ras genes are responsible for up to 30% of human tumor mutations and are composed of three isoforms: H-Ras, K-Ras and N-Ras. The post-translational modification of the CAAX motif of the Ras protein is essential in Ras actions. In the present study, we studied the effects of novel farnesyl transferase inhibitors (FTIs), YH3938 and YH3945, on the actions of oncogenic mutants of H-Ras, K-Ras and N-Ras. YH3938 and YH3945 completely reverted the proliferation and morphology of oncogenic H-Ras-transformed Rat2 cells, but not of oncogenic K-Ras-transformed Rat2 cells. Oncogenic N-Rastransformed Rat2 cells were slightly affected. Activation of SRE promoters by oncogenic H-Ras and N-Ras, but not by K-Ras, were inhibited by treatment with YH3938 and YH3945. Using bandshift analysis, YH3938 suppressed the processing of oncogenic H-Ras and N-Ras, but not that of oncogenic K-Ras protein. YH3945 only inhibited the processing of H-Ras. From these results, we conclude that YH3938 and YH3945 specifically inhibit actions of oncogenic H-Ras through inhibition of its farnesylation, that YH3938 also inhibits N-Ras activity in a dose-dependent manner, and that these drugs have no effect on oncogenic K-Ras activity.

레지오넬라 폐렴의 진단용 바이오마커의 발굴 : A/J 마우스 감염 모델에서 Legionella pneumophila의 독력 유전자들의 발현양상 분석

김승민,심희선,김희남,심호기,윤영경,김정연,박윤선,박대원,손장욱,김민자 대한감염학회 2010 감염과 화학요법 Vol.42 No.1

Background: Legionella pneumophila is the causative agent of Legionnaires’ disease, a severe form of pneumonia. After L. pneumophila is inhaled through contaminated aerosols, it is phagocytized by alveolar macrophages, multiplies in a specialized phagosome approximately 10 h postinfection, and eventually leads to the death of host cells. Currently available diagnostic tests for Legionella pneumonia have some limitations. This study was conducted to find diagnostic biomarkers for Legionella pneumonia using virulence gene expression profiling in a murine experimental model. Materials and Methods: A/J mice were intranasally inoculated with L. pneumophila serogroup 1, and lungs were harvested 4, 8, 24, and 48 h postinfection. The strain grown in buffered yeast extract broth was used as reference samples. Cy-dye labeled cDNA samples were prepared with total RNA from lungs or broth culture, and hybridized on the oligo-microarray slide containing 2,895 genes of L. pneumophila serogroup 1. Virulence gene expression patterns were analyzed using a MIDAS software from TIGR (www.tigr.org). Results: Among a total of 332 virulence genes examined, 17 genes including sidA, lepB, the genes related to flagella assembly (fliR and fliP), LPS lipid A biosynthesis, and the enhanced entry protein EnhA were up-regulated at all four time points. We further confirmed by quantitative real-time reverse transcription PCR that the expression of fliP gene was highly expressed in lung tissue as well as in bronchoalveolar lavage fluids from the mouse infected with L. pneumophila serogroup 1. Conclusions: Through gene expression analysis of L. pneumophila in a mouse model, several candidate biomarkers for diagnosing Legionnaires’ disease could be identified.

정상 성인 요통 및 요추간판탈출증 환자에서의 H-Reflex에 대한 고찰 : In Normal Dault, Lambago and Lumbar Disc Patients

이화룡,김헌주,이상철,이규호,김달수 대한신경외과학회 1980 Journal of Korean neurosurgical society Vol.9 No.1

Since the early discovery of H-reflex by Hoffmann in 1918 there have been a great number of articles in the literature suggesting its clinical uses by Magladery in the early 1950. Our experiments about about measurement of latent time in H-reflex were performed on 40 normal male adult (composed of medicos and medical officers) and 82 patients (lumbago40, lumbar disc 42) admitted at CAPGH from January '79. to August '79. The result about normal adult are as follows: 1) The mean value and the standard deviation of the latent time in normal adult (height 171±5.14cm) is 27.74 ± 1.86 maec at rihht leg, 27.77±1.64 msec at left one respectively. 2) There may be a linear relationship between latency and the height of normal adult and the difference between right and left is revealed 0.15±0.19 msec. We compare the measurement on the 82 patients group with the normal adult experimental groups in addition literatures review.

증기발생기 세관에 대한 유도 초음파의 실험적 연구

김영환,송성진,엄흥섭,김재희,김준영 한국비파괴검사학회 2003 학술대회 논문집 Vol.- No.1

증기발생기 세관 비파괴 검사에 유도초음파를 적용하기 위한 예비 실험을 수행하였다. 이를 위하여 증기 발생기 세관을 실험 대상으로 하여 분산 선도 및 특정 모드에 대응하는 초음파 입사각의 계산을 수행하였고 short time Fourier transform을 이용한 시간-주파수 분석을 통하여 유도초음파 모드를 확인하였으며, 유도 초음파가 증기발생기 세관의 곡관 부분을 통과할 때 모드 변환이 발생하지 않는 것을 실험적으로 확인하였다. 탐촉자 사이의 거리를 바꾸면서 수신한 초음파 신호에 대한 시간-주파수분석으로부터 군속도를 계산하였다. 유도 초음파를 이용한 증기발생기 세관의 최적 검사 모드를 제안하고 실험에 의하여 이를 확인하였다.

Intranasal Adenovirus-Vectored Vaccine for Induction of Long-Lasting Humoral Immunity-Mediated Broad Protection against Influenza in Mice

Kim, Eun Hye,Park, Hae-Jung,Han, Gye-Yeong,Song, Man-Ki,Pereboev, Alexander,Hong, Jeong S.,Chang, Jun,Byun, Young-Ho,Seong, Baik Lin,Nguyen, Huan H. American Society for Microbiology 2014 Journal of virology Vol.88 No.17

<P>Influenza vaccines aimed at inducing antibody (Ab) responses against viral surface hemagglutinin (HA) and neuraminidase (NA) provide sterile immunity to infection with the same subtypes. Vaccines targeting viral conserved determinants shared by the influenza A viruses (IAV) offer heterosubtypic immunity (HSI), a broad protection against different subtypes. We proposed that vaccines targeting both HA and the conserved ectodomain of matrix protein 2 (M2e) would provide protection against infection with the same subtype and also HSI against other subtypes. We report here that single intranasal immunization with a recombinant adenovirus (rAd) vector encoding both HA of H5 virus and M2e (rAdH5/M2e) induced significant HA- and M2e-specific Ab responses, along with protection against heterosubtypic challenge in mice. The protection is superior compared to that induced by rAd vector encoding either HA (rAdH5), or M2e (rAdM2e). While protection against homotypic H5 virus is primarily mediated by virus-neutralizing Abs, the cross-protection is associated with Abs directed to conserved stalk HA and M2e that seem to have an additive effect. Consistently, adoptive transfer of antisera induced by rAdH5/M2e provided the best protection against heterosubtypic challenge compared to that provided by antisera derived from mice immunized with rAdH5 or rAdM2e. These results support the development of rAd-vectored vaccines encoding both H5 and M2e as universal vaccines against different IAV subtypes.</P><P><B>IMPORTANCE</B> Current licensed influenza vaccines provide protection limited to the infection with same virus strains; therefore, the composition of influenza vaccines has to be revised every year. We have developed a new universal influenza vaccine that is highly efficient in induction of long-lasting cross-protection against different influenza virus strains. The cross-protection is associated with a high level of vaccine-induced antibodies against the conserved stalk domain of influenza virus hemagglutinin and the ectodomain of matrix protein. The vaccine could be used to stimulate cross-protective antibodies for the prevention and treatment of influenza with immediate effect for individuals who fail to respond to or receive the vaccine in due time. The vaccine offers a new tool to control influenza outbreaks, including pandemics.</P>