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Lazarte, Jassy Mary S.,Kim, Young Rim,Lee, Jung Seok,Im, Se Pyeong,Kim, Si Won,Jung, Jae Wook,Kim, Jaesung,Lee, Woo Jai,Jung, Tae Sung Elsevier 2017 Fish & shellfish immunology Vol.62 No.-
<P><B>Abstract</B></P> <P>The use of molecular adjuvants to improve the immunogenicity of DNA vaccines has been thoroughly studied in recent years. Glycoprotein (G)-based DNA vaccines had been proven to be effective in combating infection against Rhabdovirus (especially infectious hematopoietic necrosis virus, IHNV) in salmonids. DDX41 is a helicase known to induce antiviral and inflammatory responses by inducing a type I IFN innate immune response. To gain more information regarding G-based DNA vaccines in olive flounder <I>(Paralicthys olivaceus)</I>, we tried to develop a more efficient G-based DNA vaccine by adding a molecular adjuvant, DDX41. We designed a DNA vaccine in which the VHSV glycoprotein (G-protein) and DDX41 were driven by the EF-1α and CMV promoters, respectively. Olive flounders were intramuscularly immunized with 1 μg of plasmids encoding the G-based DNA vaccine alone (pEF-G), the molecular adjuvant alone (pEF-D), or the vaccine-adjuvant construct (pEF-GD). At two different time points, 15 and 30 days later, the fish were intraperitoneally infected with VHSV (100 μL; 1 × 10<SUP>6</SUP> TCID<SUB>50</SUB>/mL). Our assays revealed that the plasmid constructs showed up-regulated expression of IFN-1 and its associated genes at day 3 post-vaccination in both kidney and spleen samples. Specifically, pEF-GD showed statistically higher expression of immune response genes than pEF-G and pEF-D treated group (p < 0.05/p < 0.001). After VHSV challenge, the fish group treated with pEF-GD showed higher survival rate than the pEF-G treated group, though difference was not statistically significant in the 15 dpv challenged group however in the 30 dpv challenged group, the difference was statistically significant (p < 0.05). Together, these results clearly demonstrate that DDX41 is an effective adjuvant for the G-based DNA vaccine in olive flounder. Our novel findings could facilitate the development of more effective DNA vaccines for the aquaculture industry.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The adjuvant effect of DDX41 was assessed in this study. </LI> <LI> Simultaneous expression of VHSV glycoprotein and DDX41 showed enhanced IFN-mediated immune response. </LI> <LI> The vaccine-adjuvant construct showed enhanced regulation of type I interferon and IFN-related genes. </LI> </UL> </P>
GLOBAL EYE 국제표준화 칼럼 - ISO 표준 제정의 무대 뒤에서 -표준 제정 위해 연간 하루 평균 13차례 기술위원회회의 개최
Lazarte, Mria 기술표준원 2011 기술표준 Vol.115 No.-
ISO 시스템은 생활의 모든 분야에서 표준이 필요할 때 이해관계자들과 의견 조율을 통해 글로벌 합의를 이끌어내는 과정을 거친다.
Yogurt: Effect on Leukocytes and Blood Coagulation in an Acute Liver Injury Model
Haro, Cecilia,Lazarte, Sandra,Zelaya, Hortensia,Alvarez, Susana,Aguero, Graciela The Korean Society of Food Science and Nutrition 2009 Journal of medicinal food Vol.12 No.4
This study determined whether cow or goat yogurt administration has a preventive effect on the hepatic damage undergone during an acute liver injury. Acute liver injury was induced by an intraperitoneal injection of d-galactosamine. Groups of mice were fed with cow or goat yogurt for 2 days or 7 days before the d-galactosamine injection. Blood and liver samples were obtained 12 hours after d-galactosamine inoculation. d-Galactosamine induced an increase in serum aminotransaminases, a reduction in the number of blood leukocytes, an enhancement in neutrophil myeloperoxidase activity, a recruitment of leukocytes toward the liver, an increase in cell death, and an alteration in prothrombin time, activated partialthromboplastin time, and fibrinogen levels. Treatment with cow or goat yogurt was effective at increasing leukocyte number and decrease myeloperoxidase activity. We also observed a decrease in leukocyte accumulation in the liver and a reduction in cell death. Activated partial thromboplastin time and fibrinogen were normalized, but prothrombin time only showed an improvement without reaching normal values. Cow or goat yogurts were effective at protecting against an experimental acute liver injury, especially when administered for 7 days.
Yogurt: Effect on Leukocytes and Blood Coagulation in an Acute Liver Injury Model
Cecilia Haro,Sandra Lazarte,Hortensia Zelaya,Susana Alvarez,Graciela Agüero 한국식품영양과학회 2009 Journal of medicinal food Vol.12 No.4
This study determined whether cow or goat yogurt administration has a preventive effect on the hepatic damage undergone during an acute liver injury. Acute liver injury was induced by an intraperitoneal injection of d-galactosamine. Groups of mice were fed with cow or goat yogurt for 2 days or 7 days before the d-galactosamine injection. Blood and liver samples were obtained 12 hours after d-galactosamine inoculation. d-Galactosamine induced an increase in serum aminotransaminases, a reduction in the number of blood leukocytes, an enhancement in neutrophil myeloperoxidase activity, a recruitment of leukocytes toward the liver, an increase in cell death, and an alteration in prothrombin time, activated partial thromboplastin time, and fibrinogen levels. Treatment with cow or goat yogurt was effective at increasing leukocyte number and decrease myeloperoxidase activity. We also observed a decrease in leukocyte accumulation in the liver and a reduction in cell death. Activated partial thromboplastin time and fibrinogen were normalized, but prothrombin time only showed an improvement without reaching normal values. Cow or goat yogurts were effective at protecting against an experimental acute liver injury, especially when administered for 7 days.
Lee, Jung Seok,Kim, Jaesung,Im, Se Pyeong,Kim, Si Won,Lazarte, Jassy Mary S.,Jung, Jae Wook,Gong, Tae Won,Kim, Young Rim,Lee, Jeong Ho,Kim, Hyoung Jun,Jung, Tae Sung Elsevier 2018 Molecular immunology Vol.99 No.-
<P><B>Abstract</B></P> <P>Variable lymphocyte receptors B (VLRBs) are non-immunoglobulin components of the humoral immune system in jawless vertebrates including hagfish (<I>Eptatretus burgeri</I>) and lamprey (<I>Petromyzon marinus</I>). Hagfish VLRBs consist of leucine rich repeat (LRR) modules with a superhydrophobic C-terminal tail, the latter of which leads to extremely low expression levels in recombinant protein technology. Here, we present an artificially oligomerized VLRB (arVLRB) that conjugates <I>via</I> the C4bp oligomerization domain derived from human C4b-binding protein (hC4bp) rather than the superhydrophobic tail. The resulting arVLRB had a tightly multimerized form with seven monomeric VLRB arms and showed high expression and secretion levels in a mammalian expression system. To isolate antigen-specific arVLRB, we constructed large VLRB libraries from hagfish immunized with the fish pathogen, viral hemorrhagic septicemia virus (VHSV). The selected arVLRBs were found to recognize various types of antigens, including the recombinant target protein, purified viruses, and progeny viruses, with high antigen binding abilities and specificities. We also performed <I>in vitro</I> affinity maturation of the arVLRBs through LRRCT mutagenesis, and found that this enhanced their antigen-binding properties by at least 125-fold. Our epitope mapping analysis revealed that <SUP>37</SUP>DWDTPL<SUP>42</SUP>, which is located in a region conserved among the glycoproteins of all VHSV isolates, is the recognition epitope of the arVLRBs. Thus, our newly developed arVLRB could prove useful in the development of universal diagnostic tools and/or therapeutic agents for the virus. Together, our novel findings provide valuable insights into hagfish VLRB and its potential use as a novel alternative to conventional antibodies for biotechnological applications.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Superhydrophobic C-termini of hagfish VLRB leads to extremely low expression level. </LI> <LI> C4bp oligomerization domain mediates heptameric VLRB with high binding ability and producttivity. </LI> <LI> <I>In vitro</I> affinity maturation was efficiently carried out by LRRCT mutagenesis. </LI> <LI> Fine epitope mapping revealed <SUP>37</SUP>DWDTPL<SUP>42</SUP> is the recognition epitope of selected arVLRBs. </LI> <LI> The resulting arVLRBs can be used as diagnostic tools or therapeutic agents of VHSV. </LI> </UL> </P>