• Title/Summary/Keyword: Non-viral

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Development of a diagnostic method for human enteric Adenovirus-41 with rapid, specific and high sensitivity using the loop-mediated isothermal amplification assay

  • Lee, Jin-Young;Rho, Jae Young
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.673-681
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    • 2020
  • Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.

Clinical Study on 1 Case of Patient with Arthalgia Syndrome Diagnosed as Acute Transverse Myelitis (급성 횡단성 척수염으로 진단된 비증 환자의 치험 1례)

  • Lee, Seung-Hyun;Phil, Kam-Heun;Jo, Eun-Heui
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1663-1669
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    • 2007
  • Acute transverse myelitis(ATM) is defined as an acute intramedullary dysfunction of the spinal cord, ascendng or static involving both halves of the cord and appearing without any history of previous neurological diseases due to traumatic accident, tumor of all kind, encephalitis and of course excluding all possible viral, bacterial and fungal infection. It is mainly characterized by acute motor disorder of both limbs in respect to which spinal segments are affected as well as sensory disorder and dysuria & dyschezia. The exact cause is unknown, however it is recently suggested that immunological factors are highly involved. It has been reported by several reliable sources that it is often accompanied by immunological diseases such as systemic lupus erythematosus(SLE). As treatments non steroid anti-inflammatory drugs(NSAIDS) are primarily recommended as to steroids, limited doses are injected only with the proper prescription from the physician. Operative methods are not options as traumatic accidents and tumors are excluded as factors. To enhance muscle strength and prevent articular contracture physical therapy and passive exercise is imperative. The following patient whose chief complaints were mainly about hypoesthesia of Rt. lower limb and stiffness of phalanges of both fingers as well as to weakness of lower extermity. Therefore it has been diagnosed as arthalgia syndrome. In oriental medicine factors such as wind evil heat-evil, dampness-heat evil, cold evil cause the arthalgia syndrome. In this case the patient was diagnosed as dampness-heat evil and herbal medicine Chunglijagam-Tang and Dong-Qi acupuncture was applied to treat bladder disorder.

Preparation and In Vitro Release of DNA-Loaded Poly(D,L-lactic-co-glycolic acid) Microspheres (DNA가 봉입된 Poly(D,L-lactic-co-glycolic acid) 미립구의 제조 및 시험관내 방출)

  • Son, Hye-Jung;Kim, Jin-Seok
    • Polymer(Korea)
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    • v.29 no.1
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    • pp.69-73
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    • 2005
  • To overcome the main disadvantages of non-viral gene delivery systems such as repeated administration due to the low transfection efficiency, poly(D,L-lactide-co-glycolide) was applied to encapsulate pDNA in its microsphere formulation. Free pDNA or various ratios (w/w) of chitosan/pDNA complexes was used for encapsulation, with the resulting encapsulation efficiency of 44%, 5%, and 8% for free pDNA, 0.7:1 and 1:1 ratios, respectively. Scanning electron micrographs of poly(D,L-lactic-co-glycolic acid) (PLGA) microspheres encapsulating pDNA or chitosan-condensed pDNA revealed a smooth spherical shape immediately after microsphere preparation and a collapsed porous shape in 41 days due to the degradation of PLGA. In vitro release profile showed that the 0.7:1 (w/w) ratio formulation exerted 47% release in 26 days, whereas free pDNA or 1:1 (w/w) ratio formulation did only 15% or 32%, respectively.

Generation of Antibodies Against Rice stripe virus Proteins Based on Recombinant Proteins and Synthetic Polypeptides

  • Lian, Sen;Jonson, Miranda Gilda;Cho, Won-Kyong;Choi, Hong-Soo;Je, Yeon-Ho;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.27 no.1
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    • pp.37-43
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    • 2011
  • Rice stripe virus (RSV) is one of serious epidemic pathogens for rice species grown in many Asian countries. Therefore, it is necessary to produce a diagnostic detection kit applicable in fields for RSV detection. In this study, RSV proteins that were derived from recombinant proteins and synthetic polypeptides as antigens were generated and were raised in rabbits for antiserum production. Among seven proteins in RSV, genes that code for NCP and NS3 proteins were cloned and subcloned into vector carrying His-tag protein and were expressed in E. coli. Of two recombinant proteins, only anti-NCP displayed stable hybridization signals in western blot analysis. Alternately, synthetic RSV polypeptides for CP, NCP, NS3 and NSvc4 we also generated and only antibodies against CP and NCP were very effective to detect RSV in both RSV infected rice and weed plants. However, antibodies against NS3 and NSvc4 showed weak specific bands as well as strong non-specific background due to the difference of viral proteins produced in the infected leaves. In summary, the antibodies generated against RSV proteins produced in this study will be useful for various assays such as for RSV diagnostic detection, immunoprecipitation, protein purification, and western blot analysis.

Molecular cloning and expression analysis of an interferon stimulated gene 15 from rock bream Oplegnathus fasciatus

  • Kim, Ju-Won;Kwon, Mun-Gyeong;Park, Myoung-Ae;Hwang, Jee-Youn;Park, Hyung-Jun;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Chan-Il
    • Journal of fish pathology
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    • v.23 no.2
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    • pp.177-187
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    • 2010
  • The Interferon stimulated gene 15 (ISG15) is strongly induced in many cell types by IFNs, viral infections, and double-stranded RNA (poly I:C). The ISG15 homologue cDNA was isolated from the rock bream LPS stimulated leukocyte cDNA library. The rock bream ISG15 homologue was found to consist of 833 bp encoding 157 amino acid residues. Compared with other known ISG15 peptide sequences, the most conserved regions of the rock bream ISG15 peptide were found to be the tandem ubiquitin-like domains and a C-terminal LRLRGG conjugating motif, characteristic of mammalian and non-mammalian ISG15 proteins. Phylogenetic analysis based on the deduced amino acid sequence revealed a homologous relationship between the ISG15 sequence of rock bream and that of Atlantic salmon, Atlantic cod, northern snake head, black rockfish and olive flounder. The expression of the rock bream ISG15 molecule was induced in the peripheral blood leukocytes (PBLs) from 1 to 24 h following poly I:C stimulation, with a peak at 3 h post-stimulation. The rock bream ISG15 gene was predominantly expressed in the PBLs, spleen and gill.

A Case Report on Oro-Facial Manifestations in Leukemia (Oral Chronic GVHD) (백혈병 환자의 구강악안면 증상 발현에 관한 증례보고 (Oral Chronic GVHD))

  • Ahn, Hyoung-Joon;Kwon, Byung-Ki;Shin, Kyoung-Jin;Choi, Jong-Hoon;Kim, Chong-Youl
    • Journal of Oral Medicine and Pain
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    • v.25 no.2
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    • pp.159-165
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    • 2000
  • Subsequent to an allogenic stem cell transplantation(ASCT) on patients with hematologic malignancy(AML, ALL, CML, multiple myeloma, lymphoma etc.), chronic GVHD(graft versus host disease), which is an immunological reaction, occurs. With treatment results from patients who were diagnosed with ALL(acute lymphocytic leukemia), undergone BMT(bone marrow transplantation) and showed oral and skin lesions due to GVHD, treatment of oral manifestations of leukemia and its general management were studied. 90% of patients with chronic GVHD show change in the oral mucosa causing oral manifestations such as leukoplakia, lichenoid change of the oral mucosa, mucosal atrophy, erythema, ulceration and xerostomia. In treating GVHD, extensive systemic immunosuppression cause bacterial, viral, fungal infection that are fatal, and even if the treatment is successful, the patient is already in a severe immunosuppressed state. Therefore, localized target therapy is preferred. In another words, topical application(rinse, cream, ointment etc.) of cyclosporin and steroid in treating oral chronic GVHD is highly recommended, and the use of PUVA(Psoralen Ultraviolet A) and thalidomide is reported to be effective. In treating such diseases, dental treatment to control pain and prevent secondary infection of oral manifestations is very important. To those patients with systemic diseases who show limited effect by general dental treatment, non-invasive treatment such as the dental laser, in addition to the use of drugs, may be necessary to actively treat pain and help the healing process. For greater results, new effective methods are to be developed for treatment.

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MicroRNAs in Autoimmune Sjögren's Syndrome

  • Cha, Seunghee;Mona, Mahmoud;Lee, Kyung Eun;Kim, Dong Hee;Han, Kyudong
    • Genomics & Informatics
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    • v.16 no.4
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    • pp.19.1-19.11
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    • 2018
  • MicroRNAs (miRNAs), small non-coding RNAs, have been implicated in various diseases and cellular functions as microregulators of gene expression. Although the history of miRNA investigation in autoimmune $Sj{\ddot{o}}gren^{\prime}s$ syndrome (SjS) is fairly short, a substantial amount of data has already been accumulated. These findings clearly indicate potential clinical implications of miRNAs, such as autoantigen expression and autoantibody production, viral miRNAs regulating the calcium signaling pathway, and aberrant immune cell regulation and cytokine production. Research endeavors in the field are currently underway to select disease-specific diagnostic and prognostic biomarkers by utilizing different types of tissues or biological specimens of SjS patients. Various techniques for miRNA analysis with different stringencies have been applied, with the most recent one being next-generation sequencing. This review compiles and highlights differentially-expressed miRNAs in various samples collected from SjS patients and their potential implications in the pathogenesis of SjS. To facilitate the development of miRNA-targeted personalized therapy in the future, we urge more follow-up studies that confirm these findings and elucidate the immunopathological roles of differentially-expressed miRNAs. Furthermore, improved diagnostic criteria for the disease itself will minimize sampling errors in patient recruitment, preventing the generation of inconsistent data.

The prevalence of causative agents of calf diarrhea in Korean native calves

  • Chae, Jeong-Byoung;Kim, Hyeon-Cheol;Kang, Jun-Gu;Choi, Kyoung-Seong;Chae, Joon-Seok;Yu, Do-Hyeon;Park, Bae-Keun;Oh, Yeon-su;Choi, Hak-Jong;Park, Jinho
    • Journal of Animal Science and Technology
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    • v.63 no.4
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    • pp.864-871
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    • 2021
  • Infectious calf diarrhea is one of the most significant diseases of neonatal calves. This study is conducted to identify the prevalence of pathogens in calf diarrhea for 2 years. A total of 544 feces samples from Korean native beef calves were obtained to investigate selected seven pathogens causing calf diarrhea: bovine rotavirus, bovine coronavirus, Cryptosporidium parvum, bovine viral diarrhea virus, Eimeria species, Escherichia coli K99, and Salmonella species. The presence of diarrhea, the number and species of detected pathogens, and the calves' ages were analyzed using various statistical methods depending on the case. Of the 544 calves, 340 calves (62.5%) had normal feces and 204 calves (37.5%) had diarrhea. The presence of pathogens was significantly associated with diarrhea (p < 0.01) and fecal scores and the number of detected pathogens showed a significant linear trend (p < 0.001). Of the 7 target pathogens, 6 were detected in samples, but only C. parvum (p = 0.001) and bovine rotavirus (p < 0.001) were found at significantly higher rates in diarrheic calves than in non-diarrheic calves. Only Eimeria spp. showed a significant linear trend between the detection rate of the pathogen and the age groups (p < 0.05).

Chikungunya Virus nsP2 Impairs MDA5/RIG-I-Mediated Induction of NF-κB Promoter Activation: A Potential Target for Virus-Specific Therapeutics

  • Bae, Sojung;Lee, Jeong Yoon;Myoung, Jinjong
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1801-1809
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    • 2020
  • Chikungunya virus (CHIKV) was first identified in 1952 as a causative agent of outbreaks. CHIKV is transmitted by two mosquito species, Aedes aegypti and A. albopictus. Symptoms after CHIKV infection in human are typically fever and joint pain, but can also include headache, muscle pain, joint swelling, polyarthralgia, and rash. CHIKV is an enveloped single-stranded, positive-sense RNA virus with a diameter of approximately 70 nm. The pathogenesis of CHIKV infection and the mechanism by which the virus evades the innate immune system remain poorly understood. Moreover, little is known about the roles of CHIKV-encoded genes in the viral evasion of host immune responses, especially type I interferon (IFN) responses. Therefore, in the present study, we screened CHIKV-encoded genes for their regulatory effect on the activation of nuclear factor kappa B (NF-κB), a critical transcription factor for the optimal activation of IFN-β. Among others, non-structural protein 2 (nsP2) strongly inhibited melanoma differentiation-associated protein 5 (MDA5)-mediated induction of the NF-κB pathway in a dose-dependent manner. Elucidation of the detailed mechanisms of nsP2-mediated inhibition of the MDA5/RIG-I signaling pathway is anticipated to contribute to the development of virus-specific therapeutics against CHIKV infection.

Evaluation of virulence reversion of an attenuated porcine epidemic diarrhea vaccine strain by serial passages in suckling piglets

  • Da-Jeong Kim;Seung-Chai Kim;Hwan-Ju Kim;Gyeong-Seo Park;Sang Chul Kang;Won-Il Kim
    • Korean Journal of Veterinary Service
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    • v.46 no.3
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    • pp.193-202
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    • 2023
  • Porcine epidemic diarrhea is an infectious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV). Especially, when suckling piglets are infected, the mortality rate is close to 100%. PEDV is classified into G1 and G2 types based on genetic differences. The G2 type PEDV outbreak in the United States in 2013 was highly pathogenic and contagious, and it has spread worldwide and caused continuous economic losses. Most commercial vaccines used are G1 type vaccines, and existing vaccines do not fully protect piglets due to genetic differences. In this study, we evaluated the safety of the newly developed G2 type attenuated HSGP vaccine strain by inoculating it into piglets and testing whether the vaccine virus spreads to the non-vaccinated, negative pigs and whether the vaccine reverts to its virulence during serial passage experiments. Each experiment lasted for 7 days for each passage, and fecal viral titers, clinical symptoms, and weight gain were measured daily. After the experiment, necropsy was performed to measure intestinal virus titer and pathological evaluation. As a result of the first passage, no transmission of the vaccine virus to negative pigs co-housed with vaccinated pigs was observed. In addition, after four consecutive passage experiments, the clinical symptoms and small intestine lesions were gradually alleviated, and no virus was detected in the feces in the fourth passage experiment. Therefore, it was concluded that the vaccine was safe without virulence reversion in accordance with the guidelines of the current licensing authority. However, further studies are needed on the genetic changes and biological characteristics of the mutant virus that occur during successive passages of the attenuated vaccine since the replication and clinical symptoms of the virus increased until the third passage during successive passages of the vaccine virus. Based on this study, it was concluded that virulence reversion and safety evaluation of attenuated vaccines through serial passage in target animals can be useful to evaluate the safety of attenuated viruses.