• Title/Summary/Keyword: Non-viral

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Diagnostic Performance of Spin-Echo Echo-Planar Imaging Magnetic Resonance Elastography in 3T System for Noninvasive Assessment of Hepatic Fibrosis

  • Se Woo Kim;Jeong Min Lee;Sungeun Park;Ijin Joo;Jeong Hee Yoon;Won Chang;Haeryoung Kim
    • Korean Journal of Radiology
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    • v.23 no.2
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    • pp.180-188
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    • 2022
  • Objective: To validate the performance of 3T spin-echo echo-planar imaging (SE-EPI) magnetic resonance elastography (MRE) for staging hepatic fibrosis in a large population, using surgical specimens as the reference standard. Materials and Methods: This retrospective study initially included 310 adults (155 undergoing hepatic resection and 155 undergoing donor hepatectomy) with histopathologic results from surgical liver specimens. They underwent 3T SE-EPI MRE ≤ 3 months prior to surgery. Demographic findings, underlying liver disease, and hepatic fibrosis pathologic stage according to METAVIR were recorded. Liver stiffness (LS) was measured by two radiologists, and inter-reader reproducibility was evaluated using the intraclass correlation coefficient (ICC). The mean LS of each fibrosis stage (F0-F4) was calculated in total and for each etiologic subgroup. Comparisons among subgroups were performed using the Kruskal-Wallis test and Conover post-hoc test. The cutoff values for fibrosis staging were estimated using receiver operating characteristic (ROC) curve analysis. Results: Inter-reader reproducibility was excellent (ICC, 0.98; 95% confidence interval, 0.97-0.99). The mean LS values were 1.91, 2.41, 3.24, and 5.41 kPa in F0-F1 (n = 171), F2 (n = 26), F3 (n = 38), and F4 (n = 72), respectively. The discriminating cutoff values for diagnosing ≥ F2, ≥ F3, and F4 were 2.18, 2.71, and 3.15 kPa, respectively, with the ROC curve areas of 0.97-0.98 (sensitivity 91.2%-95.9%, specificity 90.7%-99.0%). The mean LS was significantly higher in patients with cirrhosis (F4) of nonviral causes, such as primary biliary cirrhosis (9.56 kPa) and alcoholic liver disease (7.17 kPa) than in those with hepatitis B or C cirrhosis (4.28 and 4.92 kPa, respectively). There were no statistically significant differences in LS among the different etiologic subgroups in the F0-F3 stages. Conclusion: The 3T SE-EPI MRE demonstrated high interobserver reproducibility, and our criteria for staging hepatic fibrosis showed high diagnostic performance. LS was significantly higher in patients with non-viral cirrhosis than in those with viral cirrhosis.

Human Organic Anion Transporting Polypeptide 1B3 Applied as an MRI-Based Reporter Gene

  • Song-Ee Baek;Asad Ul-Haq;Dae Hee Kim;Hyoung Wook Choi;Myeong-Jin Kim;Hye Jin Choi;Honsoul Kim
    • Korean Journal of Radiology
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    • v.21 no.6
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    • pp.726-735
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    • 2020
  • Objective: Recent innovations in biology are boosting gene and cell therapy, but monitoring the response to these treatments is difficult. The purpose of this study was to find an MRI-reporter gene that can be used to monitor gene or cell therapy and that can be delivered without a viral vector, as viral vector delivery methods can result in long-term complications. Materials and Methods: CMV promoter-human organic anion transporting polypeptide 1B3 (CMV-hOATP1B3) cDNA or CMV-blank DNA (control) was transfected into HEK293 cells using Lipofectamine. OATP1B3 expression was confirmed by western blotting and confocal microscopy. In vitro cell phantoms were made using transfected HEK293 cells cultured in various concentrations of gadoxetic acid for 24 hours, and images of the phantoms were made with a 9.4T micro-MRI. In vivo xenograft tumors were made by implanting HEK293 cells transfected with CMV-hOATP1B3 (n = 4) or CMV-blank (n = 4) in 8-week-old male nude mice, and MRI was performed before and after intravenous injection of gadoxetic acid (1.2 µL/g). Results: Western blot and confocal microscopy after immunofluorescence staining revealed that only CMV-hOATP1B3-transfected HEK293 cells produced abundant OATP1B3, which localized at the cell membrane. OATP1B3 expression levels remained high through the 25th subculture cycle, but decreased substantially by the 50th subculture cycle. MRI of cell phantoms showed that only the CMV-hOATP1B3-transfected cells produced a significant contrast enhancement effect. In vivo MRI of xenograft tumors revealed that only CMV-hOATP1B3-transfected HEK293 tumors demonstrated a T1 contrast effect, which lasted for at least 5 hours. Conclusion: The human endogenous OATP1B3 gene can be non-virally delivered into cells to induce transient OATP1B3 expression, leading to gadoxetic acid-mediated enhancement on MRI. These results indicate that hOATP1B3 can serve as an MRI-reporter gene while minimizing the risk of long-term complications.

Nuclear Imaging Evaluation of Galactosylation of Chitosan (핵의학 영상을 이용한 chitosan의 galactosylation 효과에 대한 평가)

  • Jeong, Hwan-Jeong;Kim, Eun-Mi;Park, In-Kyu;Cho, Chong-Su;Kim, Chang-Guhn;Bom, Hee-Seung
    • The Korean Journal of Nuclear Medicine
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    • v.38 no.3
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    • pp.253-258
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    • 2004
  • Purpose: Chitosan has been studied as a non-viral gene delivery vector, drug delivery carrier, metal chelator, food additive, and radiopharmaceutical, among other things. Recently, galactose-graft chitosan was studied as a non-viral gene and drug delivery vector to target hepatocytes. The aim of this study was to investigate the usefulness of nuclear imaging for in vivo evaluation of targeting the hepatocyte by galactose grafting. Methods and Materials: Galactosyl methylated chitosan (GMC) was produced by methylation to lactobionic acid coupled chitosan. Cytotoxicity of $^{99m}Tc$-GMC was determined by MTT assay. Rabbits were injected via their auricular vein with $^{99m}Tc$-GMC and $^{99m}Tc$-methylated chitosan (MC), the latter of which does not contain a galactose group, and images were acquired with a gamma camera equipped with a parallel hole collimator. The composition of the galactose group in galactosylated chitosan (GC), as well as the tri-, di-, or mono-methylation of GMC, was confirmed by NMR spectroscopy. Results: The results of MTT assay indicated that $^{99m}Tc$-GMC was non-toxic. $^{99m}Tc$-GMC specifically accumulated in the liver within 10 minutes of injection and maintained high hepatic uptake. In contrast, $^{99m}Tc$-MC showed faint liver uptake. $^{99m}Tc$-GMC scintigraphy of rabbits showed that the galactose ligand principally targeted the liver while the chitosan functionalities led to excretion through the urinary system. Conclusion: Bioconjugation with a specific ligand endows some degree of targetability to an administered molecule or drug, as in the case of galactose for hepatocyte in vivo, and evaluating said targetabililty is a clear example of the great benefit proffered by nuclear imaging.

Effects on Growth and Yield of Korean Malting Barley Cultivars by Soil-borne Bymovirus Infection (맥류의 토양 전염성 Bymovirus의 감염이 국내 육성 맥주보리의 생육 및 수량에 미치는 영향)

  • Park, Jong-Chul;Kim, Mi-Jung;Lee, Eun-Sook;Park, Chul-Soo;Kang, Chun-Sik;Hyun, Jong-Nae;Lee, Jung-Joon;Kim, Tae-Soo;Kim, Ki-Jong
    • Research in Plant Disease
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    • v.16 no.1
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    • pp.21-26
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    • 2010
  • Viral diseases, especially Barley yellow mosaic virus (BaYMV) and Barley mild mosaic virus (BaMMV) have been most serious in barley fields. In this study, we investigated the effect of different level of resistance to viral diseases on the plant growth and yield in malting barley. In diagnosis of virus infection, BaYMV and BaMMV were detected in 'Doosan 29' (susceptible), however, 'Jinyangbori' (moderate susceptible) and 'Hopumbori' (moderate) was infected by only BaYMV. Plant height was restrained about 8~29% in overwintered plant regeneration stage depending on the resistant of each cultivar. The culm length damaged also to 9~12% by BaYMV infection. The tiller numbers reduced to 10~14% in overwintering season, however, the head numbers in harvest season more decreased to 26~33%. Heading date was delayed to 3~3 days by the infection. In examination of yield components, 1,000 kernel weight and $\ell$ weight reduced according to culrivar's resistant degrees to 4.0~6.4% and 1.0~4.2%, respectively. The yield of abortive grain was doubled in BaYMV infection comparing to non-infested field. Three varieties tested in the non-infected field over two years were not significantly different for yield potential with ranges of 509kg~632kg/10a. However, significant yield reduction was observed in 'Saessalbori' and 'Baegdong' with ranges of 77~177kg/10a as compared to 'Hopumbori' (467 kg/10a) when tested in the virus-infected field. Yield potentials of these cultivars reduced by 26~43%, respectively, in the virus-infected field as compared to those in the non-infected field.

Development of a Duplex RT-PCR Assay for the Simultaneous Detection and Discrimination of Avirulent and Virulent Newcastle Disease Virus (NDV) (뉴캣슬병 바이러스 검출 및 병원성 감별을 위한 Duplex RT-PCR법 개발)

  • Kim, Ji-Ye;Lee, Hyun-Jeong;Jang, Il;Lee, Hee-Soo;Yoon, Seung-Jun;Park, Ji-Sung;Seol, Jae-Goo;Kim, Seung-Han;Hong, Ji-Mu;Wang, Zillian;Liu, Hualei;Choi, Kang-Seuk
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.93-102
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    • 2017
  • A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.

Evaluation for Protective Effect of CPV-2 and CPV-2b Vaccines against a Korean CPV-2a Isolate in Pups (국내에서 유행하는 CPV-2a 분리주에 대한 CPV-2와 CPV-2b 백신의 방어효능 평가)

  • Jeoung, Seok-Young;Yi, Jun-Seok;Kim, Hyun-Tae;Kim, Doo
    • Journal of Veterinary Clinics
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    • v.31 no.5
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    • pp.361-366
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    • 2014
  • The aim of this study was to determine if vaccines containing CPV-2 or CPV-2b provided protection against challenge with a recent Korean CPV-2a isolate. Twenty mongrel pups aged 9 weeks old were used. The commercial CPV-2 or CPV-2b vaccines were administered to each of the 8 pups thrice every 3 weeks, respectively. Two weeks after the last vaccination, all pups were challenged with CPV-2a (VR00174 strain) $1{\times}10^6\;TCID_{50}$. Clinical signs, fecal excretion of challenged CPV, and serological response of pups were observed for 2 weeks after challenge. All vaccinated pups did not display any clinical signs of disease after challenge with Korean CPV-2a isolate, whereas all non-vaccinated pups exhibited mucoid or hemorrhagic diarrhea, vomiting and anorexia. In all non-vaccinated pups, the virus could be detected in feces from 4 days after challenge, whereas in vaccinated pups, no evidence of viral excretion could be detected. Two of 4 non-vaccinated pups died 6 days after the challenge. This study showed that the two commercial CPV-2 and CPV-2b vaccines were effective in preventing infection and/or disease caused by the Korean CPV-2a isolate.

Characteristics of Meningitis with or without Enterovirus (엔테로바이러스의 유무에 따른 수막염의 특성)

  • Cho, Won Je;Kwon, Ye Rim;Cha, Byung Ho
    • Journal of the Korean Child Neurology Society
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    • v.26 no.4
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    • pp.246-250
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    • 2018
  • Purpose: Meningitis is an acute childhood infection caused by viral or bacterial infection. The purpose of the present study is to analyze the differences between enteroviral meningitis and non-enteroviral aseptic meningitis. Methods: From January 2013 to December 2016, we retrospectively reviewed the medical records of a total of 303 aseptic meningitis patients who visited Wonju Severance Christian Hospital. We examined demographics of all patients and analyzed serologic and cerebrospinal fluid tests, clinical symptoms, and outcomes. Results: Of a total of 303 patients, 197(65.0%) were male, and the most cases occurred from June to November (91.8%). The most common pathogen of meningitis was found to be enterovirus (65.0%). According to the etiology, the enteroviral meningitis group had significantly more headache and enteric symptoms (P=0.0003 and P=0.0013, respectively). Furthermore, the non-enteroviral meningitis group showed pleocytosis in the cerebrospinal fluid and a significantly higher rate of seizure at 1 to 4 years (P=0.0360 and P=0.0002, respectively). Conclusion: In this study, enteroviral meningitis was the most common and the prognosis was good. When compared between two groups, neurological symptoms were frequent in patients with non-enteroviral meningitis in groups 1 to 4 years.

Effect of severe acute respiratory syndrome coronavirus 2 infection during pregnancy in K18-hACE2 transgenic mice

  • Byeongseok, Kim;Ki Hoon, Park;Ok-Hee, Lee;Giwan, Lee;Hyukjung, Kim;Siyoung, Lee;Semi, Hwang;Young Bong, Kim;Youngsok, Choi
    • Animal Bioscience
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    • v.36 no.1
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    • pp.43-52
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    • 2023
  • Objective: This study aimed to examine the influence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection on pregnancy in cytokeratin-18 (K18)-hACE2 transgenic mice. Methods: To determine the expression of hACE2 mRNA in the female reproductive tract of K18-hACE2 mice, real-time polymerase chain reaction (RT-PCR) was performed using the ovary, oviduct, uterus, umbilical cord, and placenta. SARS-CoV-2 was inoculated intranasally (30 μL/mouse, 1×104 TCID50/mL) to plug-checked K18-hACE2 homozygous female mice at the pre-and post-implantation stages at 2.5 days post-coitum (dpc) and 15.5 dpc, respectively. The number of implantation sites was checked at 7.5 dpc, and the number of normally born pups was investigated at 20.5 dpc. Pregnancy outcomes, including implantation and childbirth, were confirmed by comparison with the non-infected group. Tissues of infected mice were collected at 7.5 dpc and 19.5 dpc to confirm the SARS-CoV-2 infection. The infection was identified by performing RT-PCR on the infected tissues and comparing them to the non-infected tissues. Results: hACE2 mRNA expression was confirmed in the female reproductive tract of the K18-hACE2 mice. Compared to the non-infected group, no significant difference in the number of implantation sites or normally born pups was found in the infected group. SARS-CoV-2 infection was detected in the lungs but not in the female reproductive system of infected K18-hACE2 mice. Conclusion: In K18-hACE2 mice, intranasal infection with SARS-CoV-2 did not induce implantation failure, preterm labor, or miscarriage. Although the viral infection was not detected in the uterus, placenta, or fetus, the infection of the lungs could induce problems in the reproductive system. However, lung infections were not related to pregnancy outcomes.

Isolation and Evaluation of an Antiviral Producing Serratia spp. Strain Gsm01 against Cucumber mosaic virus in Korea (한국에서 CMV에 항바이러스 효과를 나타내는 Serratia spp. Gsm01 균주의 분리 동정 및 효과 검정)

  • Ipper, Nagesh S.;Lee, Seon-Hwa;Suk, Jung-Ki;Shrestha, Anupama;Seo, Dong-Uk;Park, Duck-Hwan;Cho, Jun-Mo;Park, Dong-Sik;Hur, Jang-Hyun;Lim, Chun-Keun
    • The Korean Journal of Pesticide Science
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    • v.10 no.4
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    • pp.344-350
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    • 2006
  • An Antiviral producing bacterial strain was isolated from ginseng root environment in Hongcheon, Kangwon province of Republic of Korea. Identification of this bacterial strain was performed by physiological and biochemical tests along with 16S rRNA analyses. The results revealed that the bacterium was closer to genus Serratia, which was named as Gsm01. The strain was grown in Mannitol-Glutamate-Yeast (MGY) broth for 48 h. The culture was centrifuged and the filtrate obtained was tested for its ability to control Cucumber mosaic virus strain Y (CMV-Y) in greenhouse and field experiments. In the green house experiments, CF was evaluated for its ability to protect local host, Chenopodium amaranticolor and systemic host of CMV, Nicotiana tabacum cv. Xanthi-nc. It was found that, CF treatment reduced viral infection by 98% in local host; C. amaranticolor. The N. tabacum cv. Xanthi-nc plants treated with CF did not show visible viral symptoms 15 days post inoculation (dpi) and remained symptomless throughout the periods of the study. To evaluate effectiveness of CF under field conditions, experiment was carried out in a polyvinyl house. It was observed that, 52% plants were protected from viral diseases compared to non-treated plants, increasing the crop yield. This is the first report showing antiviral activity of a Serratia spp. against CMV.

Molecular Miology of the Poliovirus (폴리오바이러스의 분자생물학)

  • 최원상
    • Journal of Life Science
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    • v.7 no.4
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    • pp.392-401
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    • 1997
  • The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.

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