• Title/Summary/Keyword: Non-viral

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Current advances in adenovirus nanocomplexes: more specificity and less immunogenicity

  • Kang, Eun-Ah;Yun, Chae-Ok
    • BMB Reports
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    • v.43 no.12
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    • pp.781-788
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    • 2010
  • An often overlooked issue in the field of adenovirus (Ad)-mediated cancer gene therapy is its limited capacity for effective systemic delivery. Although primary tumors can be treated effectively with intralesional injection of conventional Ad vectors, systemic metastasis is difficult to cure. Systemic administration of conventional naked Ads leads to acute accumulation of Ad particles in the liver, induction of neutralizing antibody, short blood circulation half-life, non-specific biodistribution in undesired organs, and low selective accumulation in the target disease site. Versatile strategies involving the modification of viral surfaces with polymers and nanomaterials have been designed for the purpose of maximizing Ad anti-tumor activity and specificity by systemic administration. Integration of viral and non-viral nanomaterials will substantially advance both fields, creating new concepts in gene therapeutics. This review focuses on current advances in the development of smart Ad hybrid nanocomplexes based on various design-based strategies for optimal Ad systemic administration.

Targeted Polymeric Gene Delivery for Anti-angiogenic Tumor Therapy

  • Kim, Won-Jong;Kim, Sung-Wan
    • Macromolecular Research
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    • v.15 no.2
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    • pp.100-108
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    • 2007
  • Gene therapy has become a promising strategy for the treatment of genetically based diseases, such as cancer, which are currently considered incurable. A major obstacle in the field of cancer gene therapy is the development of a safe and efficient delivery system for therapeutic gene transfer. Non-viral vectors have attracted great interest, as they are simple to prepare, stable, easy to modify and relatively safe compared to viral vectors. In this review, an insight into the strategies developed for polyethylenimine (PEI)-based non-viral vectors has been provide, including improvement of the polyplex properties by incorporating hydrophilic spacer, poly(ethylene glycol) (PEG). Moreover, this review will summarize the strategies for the tumor targeting. Specifically, a targeted polymeric gene delivery system, PEI-g-PEG-RGD, will be introduced as an efficient gene delivery vector for tumor therapy, including its functional analysis both in vitro and in vivo.

Clinical Experiences with Febrile Infants, Younger than Three Months, with Urinary Tract Infection in a Single Center for Four Years : Incidence, Pathogens and Viral Co-infections

  • Yoon, Seok Ho;Shin, Son Moon;Park, Sung Won
    • Childhood Kidney Diseases
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    • v.21 no.2
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    • pp.101-106
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    • 2017
  • Purpose: The aim of this study was to analyze the incidence and microbiological characteristics of urinary tract infection in infants aged younger three months and to compare with other infection with positive urine culture. Methods: We retrospectively reviewed the medical records of 425 infants with a tympanic temperature >$37.6^{\circ}C$, aged younger than three months, who were admitted to Cheil General Hospital in Seoul, Korea, from January 2013 to December 2016. Demographic and clinical features, laboratory findings, respiratory virus PCR and the pathogens of a urine culture were analyzed. Results: A total of 88 infants (63 males, 25 females) had urinary pathogens detected in the urine culture test. The incidence of UTI in febrile infants aged younger 3 months was 11%. The most common pathogen which causes UTI was E. coli as same as in previous studies. They were divided into a UTI group (n=48) and a non-UTI group (n=40). In comparison of both group, leukocytosis, C-reactive protein level, Absolute neutrophil count level, peak temperature is statistically significant. In both group, there were co-infections with viral pathogens in some cases, and the odd ratio of non-UTI group with viral infection was 3.28. Conclusion: The study determined the incidence and pathogen of UTI in febrile infants, aged younger three months. E. coli was responsible for the majority UTI. There were some viral co-infections in febrile infants with bacteriuria and incidence was higher in non-UTI group. WBC count, ANC count and CRP level were the differentiating factors of UTI from non-UTI group.

Detection of Specific Antibodies Against Viral Hemorrhagic Septicemia Virus in Infected Olive Flounder Paralichthys olivaceus Using Enzyme-Linked Immunosorbent Assay (Enzyme-linked immunosorbent assay를 이용한 바이러스성 출혈성 패혈증 바이러스 감염 넙치(Paralichthys olivaceus)의 특이 항체반응 검사)

  • Hwang, Jee Youn;Jang, Jin Hyeon;Kim, Dong Jun;Kwon, Mun Gyeong;Seo, Jung Soo;Hwang, Seong Don;Son, Maeng-Hyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.5
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    • pp.547-552
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    • 2017
  • The viral hemorrhagic septicemia virus (VHSV) has an extensive host range, and infects farmed and wild fish inhabiting both freshwater and marine ecosystems. Enzyme-linked immunosorbent assay (ELISA) is highly useful in diagnosing viral hemorrhagic septicemia. However, ELISA shows high, non-specific background reaction with fish antibodies. In this study, we optimized the antigen and antibody concentrations used for detecting specific antibodies in VHSV-infected olive flounder to reduce non-specific binding, and improve the sensitivity of ELISA. The results suggested that OD (optical Density) values were valid when ELISA was performed with $0.1{\mu}g/well$ of virus, involving blocking with blocking buffer (Roth, Roti-Block), 1:300-1:600 dilution with flounder antisera, and 1:1000 dilution with anti-flounder IgM and HRP-conjugated goat anti-mouse IgG for detecting the VHSV antibody in flounder sera. Furthermore, 11 different VHSV strains isolated in Korea from 2012 to 2016 were used to infect the fish. The results showed no correlation between viral pathogenicity and antibody production. This research is a basic study on the application of antibody detection in the diagnosis of viral hemorrhagic septicemia in the olive flounder.

Association of Viral Infections with Risk of Human Lymphomas, Egypt

  • Kadry, Dalia Y;Khorshed, Amira M;Rashed, Reham A;Mokhtar, Nadia M
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1705-1712
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    • 2016
  • Background: The aim of this study was to determine and evaluate the association of different viral infections, with hepatitis B and C viruses, Epstein-Barr virus, cytomegalovirus and human herpes virus-8 (HBV, HCV, EBV, CMV, HHV-8) with the risk of lymphomas (Hodgkin and non-Hodgkin) among Egyptian patients, and correlate with the histopathological staging and typing as well as the prevalence of combined infections. Materials and Methods: A total of 100 newly diagnosed lymphoma patients with 100 healthy age and sex matched normal controls were assayed for viral infection using enzyme linked immunosorbant assay (ELISA) followed by real time polymerase chain reaction (RT-PCR). Results: Our results showed a high statistical significant difference between cases and controls as regards clinical and laboratory findings (P<0.001 and=0.003). A high statistical difference was seen for the association of most viruses and lymphoma cases (p<0.001) except for positive HBs Ag, positive CMV IgG and HHV-8 (p=0.37, 0.70 and 1.0 respectively). No statistical significant difference was found between Hodgkin (HL) and non-Hodgkin (NHL) as regards viral prevalence except HCV antigen, 57.1% for HL and 26.5% for NHL (p = 0.03). Only, HBV DNA showed a high significant value among infiltrated bone marrow cases (p=0.003) and finally, a high significant association of 2 combined viral infections with infiltrated bone marrow lymphoma cases (p=0.04). Conclusions: Our results showed that infection with HBV, HCV, CMV and EBV were associated with increased risk of lymphoma among the Egyptian population. Detection of new associations between infectious agents and risk of cancer development will facilitate progress in elaboration of prophylactic measures, early diagnostic methods and, hopefully, novel therapy of malignant tumours.

Review on the Occurrence and Studies of Potato Viral Diseases in Korea (한국에서의 감자 바이러스병 발생과 그 연구에 대한 고찰)

  • Hahm, Young-Il
    • Research in Plant Disease
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    • v.9 no.1
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    • pp.1-9
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    • 2003
  • The occurrence of potato(Sotanum tuberosum) viral diseases caused by Potato virus X(PVX), Potato virus Y (PVY), Potato leafroll virus(PLRV), Potato vims S(PVS), Potato virus M(PVM), Potato virus A(PVA), Potato virus T(PVT), Alfalfa mosic virus(AIMV), Tobacco mosic virus(TMV), Potato mop top virus(PMTV) Tobacco rattle virus(TRV) and Potato spindle tuber viroid(PSTVd), potato witches' broom phytoplasma, have been identified so far in Korea. Major viral diseases such as PVX, PVY and PLRV had been studied more deeply, however, the others are just identified and only partially characterized since the first study on the relation between PVX nucleic acid and virus protein by Kim in 1961. The most studies on potato viral diseases are mainly focused on the problems of seed potato production. The National Alpine Agricultural Experiment Station(NAAES), since it began its activities in 1961, has given special attention to this problem by doing studies to identify, characterize and control potato virus diseases. This effort resulted in the development of new potato virus detection methods as a basis for elaborating new method of control, such as the production of seed potato free of virus and the selection of new virus-resistant transgenic potatoes. The further studies of potato viral diseases required would be fallowings: the continuous monitoring for the occurrence of identified or not identified potato viruses in Korea, the isolation of resistant viral genes, the development of control method for the non-persistently transmitted viruses like PVY, special vectors such as nematode and fungus transmitted viruses, TRV and PMTV and the development of control methods against potato viral diseases by viral cross protection, therapy, transgenic plant, and the use of the agents or molecules, such as virus inhibitors and antiviral proteins, etc., blocking viral replication.

Antibody-secreting macrophages generated using CpG-free plasmid eliminate tumor cells through antibody-dependent cellular phagocytosis

  • Cha, Eun Bi;Shin, Keun Koo;Seo, Jinho;Oh, Doo-Byoung
    • BMB Reports
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    • v.53 no.8
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    • pp.442-447
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    • 2020
  • The non-viral delivery of genes into macrophages, known as hard-to-transfect cells, is a challenge. In this study, the microporation of a CpG-free and small plasmid (pCGfd-GFP) showed high transfection efficiency, sustainable transgene expression, and good cell viability in the transfections of Raw 264.7 and primary bone marrow-derived macrophages. The non-viral method using the pCGfd vector encoding anti-EGFR single-chain Fv fused with Fc (scFv-Fc) generated the macrophages secreting anti-EGFR scFv-Fc. These macrophages effectively phagocytized tumor cells expressing EGFR through the antibody-dependent mechanism, as was proved by experiments using EGFR-knockout tumor cells. Finally, peri-tumoral injections of anti-EGFR scFv-Fc-secreting macrophages were shown to inhibit tumor growth in the xenograft mouse model.

Establishment of reverse transcription polymerase chain reaction for detection of Getah virus infection in livestock

  • Lee, Seung Heon;Yang, Dong-Kun;Kim, Ha-Hyun;Choi, Sung-Suk;Cho, In-Soo
    • Korean Journal of Veterinary Research
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    • v.57 no.1
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    • pp.37-42
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    • 2017
  • Getah virus (GETV) infection causes sporadic outbreaks of mild febrile illness in horses and reproductive failure in pigs. In this study, we established a reverse transcription polymerase chain reaction (RT-PCR) method to detect GETV from suspected virus-infected samples. The reaction conditions were optimized and validated by using RNA extracted from GETV propagated in cell culture. A GETV-specific GED4 primer set was designed and used to amplify a 177 bp DNA fragment from a highly conserved region of the E1 glycoprotein gene in the GETV genome. RT-PCR performed with this primer set revealed high sensitivity and specificity. In the sensitivity test, the GED4 primer set detected GETV RNA at the level of $10^{2.0}\;TCID_{50}/mL$. In the specificity test, the GED4 primer set amplified only a single band of PCR product on the GETV RNA template, without non-specific amplification, and exhibited no cross-reactivity with other viral RNAs. These results suggest that this newly established RT-PCR method is useful for accurate identification of GETV infection in animals.

Highly Efficient Gene Expression in Rabbit Synoviocytes Using EBV-Based Plasmid (가토 윤활막 세포에서 EBV-Based 플라스미드를 사용한 효율적인 유전자 발현)

  • Kim, Jin Young;Oh, Sang Taek;Youn, JeeHee;Lee, Suk Kyeong
    • IMMUNE NETWORK
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    • v.4 no.3
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    • pp.190-197
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    • 2004
  • Background: Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic synovial inflammation which leads to joint destruction. Gene therapy of RA targets the players of inflammation or articular destruction. However, viral vectors have safety problems and side effects, while non-viral vectors suffer from inefficient gene transfer and fast loss of gene expression. To overcome the limits of non-vial vectors, an EBV-based plasmid which is known to exert prolonged high level gene expression can be used. Methods: pEBVGFP, pEBVIL-10, and pEBVvIL-10 were constructed by cloning GFP, IL-10, and vIL-10 genes into an EBV-based plasmid, respectively. The pGFP was used as a control plasmid. Each constructs were lipofected into HIG-82 rabbit synoviocytes. The expression of GFP was monitored by FACS and confocal microscopy. IL-10 and vIL-10 expressions were measured by ELISA. Results: GFP expression 2 days after transfection was achieved in 33.2% of cells. GFP-expressing cells transfected with pGFP decreased rapidly from 4 days after transfection and disappeared completely by 11 days. Cells transfected with pEBVGFP began to decrease slowly from 4 days. But GFP expression was detected for over 35 days. In addition, HIG-82 cells transfected with pEBVIL-10 ($44.6{\pm}1.5ng/ml$) or pEBVvIL-10 ($51.0{\pm}5.7ng/ml$) secreted these cytokines at high levels. High level cytokine production by hygromycin selection was maintained at least for up to 26 days after transfection. Conclusion: These results suggest that the EBV-based plasmid has a potential to improve non-viral gene transfer system and may be applicable to treat RA without the drawbacks of viral vectors.