• 제목/요약/키워드: reproduction system

검색결과 839건 처리시간 0.028초

Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat)

  • Unsunnidhal, Lalu;Wasito, Raden;Setyawan, Erif Maha Nugraha;Warsani, Ziana;Kusumawati, Asmarani
    • Journal of Veterinary Science
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    • 제22권6호
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    • pp.76.1-76.15
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    • 2021
  • Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.

Tissues Expression, Polymorphisms of IFN Regulatory Factor 6 (IRF6) Gene and Their Associated with Immune Traits in Three Pig Populations

  • Liu, Yang;Xu, Jingeng;Fu, Weixuan;Weng, Ziqing;Niu, Xiaoyan;Liu, Jianfeng;Ding, Xiangdong;Zhang, Qin
    • Asian-Australasian Journal of Animal Sciences
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    • 제25권2호
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    • pp.163-169
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    • 2012
  • Interferon regulatory factor 6 (IRF6) gene is a member of the IRF-family, and plays functionally diverse roles in the regulation of the immune system. In this report, the 13,720 bp porcine IRF6 genomic DNA structure was firstly identified with a putative IRF6 protein of 467 amino acids. Alignment and phylogenetic analysis of the porcine IRF6 amino acid sequences with their homologies to other species showed high identity (over 96%). Tissues expression of IRF6 mRNA was observed by RT-PCR, the results revealed IRF6 expressed widely in eight tissues. One SNP (HQ026023:1383 G>C) in exon7 and two SNPs (HQ026023:130 G>A; 232 C>T) in the 5′ promoter region of porcine IRF6 gene were demonstrated by DNA sequencing analysis. A further analysis of SNP genotypes associated with immune traits including IFN-${\gamma}$ and IL10 concentrations in serum was carried out in three pig populations including Large White, Landraces and Songliao Black pig (a Chinese indigenous breed). The results showed that the SNP (HQ026023:1383 G>C) was significantly associated with the level of IFN-${\gamma}$ (d 20) in serum (p = 0.038) and the ratio of IFN-${\gamma}$ to IL10 (d 20) in serum (p = 0.041); The other two SNPs (HQ026023:130 G>A; 232 C>T) were highly significantly associated with IL10 level in serum both at the day 20 (p = 0.005; p = 0.001) and the day 35 (p = 0.004; p = 0.006). Identification of the porcine IRF6 gene will help our further understanding of the molecular basis of the IFN regulation pathway in the porcine immune response. All these results should indicate that the IRF6 gene can be regarded as a molecular marker associated with the IL10 level in serum and used for genetic selection in the pig breeding.

Retrovirus Vector를 이용한 동물 수정란에의 유전자 전이 (Retrovirus Vector-mediated Gene Transfer into the Fertilized Embryos of the Farm Animals)

  • 김태완
    • 한국가축번식학회지
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    • 제19권4호
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    • pp.293-305
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    • 1996
  • Retrovirus는 DNA가 아닌 RNA를 유전 물질로 갖고 있는 동물 virus인데 각 virus는 RNA와 함께 크게 gag, pol. 그리고 env 등의 3가지 단백질로 구성되어 있다. gag 단백질은 virus의 내부구조를 형성하는 단백질이고, pol단백질은 감염을 통해 표적 세포에 도입된 retrovirus의 RNA를 DNA로 역전사시키는 reverse transcriptase의 역할을 하며, env단백질은 virusdml 외부를 구성하는 단백질로써 이 단백질에 의해 각 retrovirus의 종류에 따른 감염이 가능한 표적세포의 종류가 결정된다(host cell specificity). 따라서 어떤 retrovirus의 envelope단백질과 표식세포에 있는 retrovirus의 envelope 단백질에 대한 특정 receptor와의 상호 작용에 의해 세포속으로 도입된 virus의 RNA는 reverse transcriptase에 의해 DNA로 역전사된 후 표적세포의 genomic DNA에 삽입되는 특징을 가진다. 이러한 특징을 가진 retrovirus vector system은 형질 전환 동물의 생산에 있어서 현재까지의 주된 방법인 수정란의 pronucleus에의 DNA microinjection방법 보다 여러 가지 면에서 우수함에도 불구하고 쥐 이외의 다른 동물에서는 거의 이용되고 있지 않는 실정이다. 주된 원인으로는 현재 사용되고 있는 대부분의 retrovirus vector system이 쥐의 백혈병 virus를 근간으로 하기 때문에 이 system에서 생산된 virus는 쥐 이외의 다른 동물, 특히 유제류의 세포에는감염성이 아주 약하기 때문이다. 이러한 결점을 해결하기 위하여 최근에 기존의 쥐 백혈병 virus의 envelope protein을 vesicular stomatitis virus의 G protein으로 대체한 hybrid retrovirus vector system이 개발되었다. 이러한 system에서 생산되는 virus는 조류를 포함한 거의 모든 종류의 동물세포를 감염시킬 수 있으며 몇몇 특정세포에 대해서는 기존의 retrovirus vector system에 비해 1,000배 이상의 높은 감염도를 나타내는데 그 특징이 있다. 따라서 이러한 새로운 virus vector system을 이용할 경우, 보다 다양한 종에 있어서 형질전환 동물을 효율적으로 생산할 수 있을 뿐만 아니라 형질전환 동물의 생산 방법 자체를 다양화 시킬 수 있다고 본다.

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Neurokinin B-related Peptide Suppresses the Expression of GnRH I, Kiss2 and tac3 in the Brain of Mature Female Nile tilapia Oreochromis niloticus

  • Jin, Ye Hwa;Park, Jin Woo;Kim, Jung-Hyun;Kwon, Joon Yeong
    • 한국발생생물학회지:발생과생식
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    • 제20권1호
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    • pp.51-61
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    • 2016
  • Neurokinin B (NKB) and neurokinin B related peptide (NKBRP) belong to tachykinin peptide family. They act as a neurotransmitter and/or neuromodulator. Mutation of NKB and/or its cognate receptor, NK3R resulted in hypogonadotropic hypogonadism in mammals, implying a strong involvement of NKB/NK3R system in controlling mammalian reproduction. Teleosts possess NKBRP as well as NKB, but their roles in fish reproduction need to be clarified. In this study, NKB and NKBRP coding gene (tac3) was cloned from Nile tilapia and sequenced. Based on the sequence, Nile tilapia NKB and NKBRP peptide were synthesized and their biological potencies were tested in vitro pituitary culture. The synthetic NKBRP showed direct inhibitory effect on the expression of GTH subunits at the pituitary level. This inhibitory effect was confirmed in vivo by means of intraperitoneal (ip) injection of synthetic NKB and NKBRP to mature female tilapia (20 pmol/g body weight [BW]). Both NKB and NKBRP had no effect on the plasma level of sex steroids, E2 and 11-KT. However, NKBRP caused declines of expression level of GnRH I, Kiss2 and tac3 mRNAs in the brain while NKB seemed to have no distinct effect. These results indicate some inhibitory roles of NKBRP in reproduction of mature female Nile tilapia, although their exact functions are not clear at the moment.

Factors affecting in vitro embryo production: insights into dromedary camel

  • Moawad, Adel R.;Ghoneim, Ibrahim M.;Darwish, Gamal M.;Badr, Magdy R.;El-Badry, Diya A.;EL-Wishy, Abou Bakr A.
    • 한국동물생명공학회지
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    • 제35권2호
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    • pp.119-141
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    • 2020
  • The Dromedary camel (Camelus dromedaries) is an important species because of its ability to produce good quality meat, milk, and fibers under harsh environmental conditions. Camels are also crucial for transportation, racing, and as draft animals in agriculture. Therefore, dromedary camels play a critical role in the economy for millions of people living in the arid part of the world. The inherent capability of camels to produce meat and milk is highly correlated with their reproductive performance. Compared with other domestic species, the reproductive efficiency in camelids is low. Although recent reproductive technologies such as in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) have been successfully applied to camelids and the birth of live offspring following these technologies has been reported; in vitro embryo production (IVP) has lagged in this species. The development of the IVP system for dromedary camels may be a useful tool for the genetic improvement of this species. IVP in farm animals includes three main steps; in vitro maturation (IVM) of an oocyte, IVF of a matured oocyte, and in vitro culture (IVC) of fertilized oocyte up to the blastocyst stage. This review aims to summarize various factors that influence oocyte quality, IVM, and in vitro embryo development in dromedary camel.

Feeding strategies alter gene expression of the calpain system and meat quality in the longissimus muscle of Braford steers

  • Coria, Maria Sumampa;Reineri, Pablo Sebastian;Pighin, Dario;Barrionuevo, Maria Guadalupe;Carranza, Pedro Gabriel;Grigioni, Gabriela;Palma, Gustavo Adolfo
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권5호
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    • pp.753-762
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    • 2020
  • Objective: The aim of the present study was to determine the effect of supplementing pasture-finished steers with corn silage on the expression level of the calpain system proteins and beef tenderization. Methods: Thirty Braford steers grazing on summer pasture were used for the study. For 120 days fifteen animals were supplemented with corn silage at 1% of body weight per head per day (Suppl) whereas the remaining 15 steers only received pasture (Contr). Carcass and meat traits were evaluated and compared between groups. Gene expression and activities of proteases (calpain 1 and calpain 2) and inhibitor (calpastatin) were measured using real-time polymerase chain reaction and casein zymography. Results: Carcass and meat traits were significantly different between feeding systems. Supplemented steers showed higher hot carcass weight (p<0.01), fat content (p = 0.02), and Warner-Bratzler shear force (p = 0.03). Furthermore, the control group showed higher protease:inhibitor ratios, at mRNA (p = 0.01) and protein levels (p<0.10). Warner-Bratzler shear force and mRNA calpains:calpastatin ratio were associated in both feeding systems (p<0.01). Conclusion: Based on the results obtained in the study, beef tenderness differences among finishing strategies could be modulated through differential expression of the calpain system proteins.

초미립자탄산칼슘이 잉크제트 인쇄품질에 미치는 영향 (The Effect of Precipitated Calcium Carbonate Having a Small Particle Size on the Print Quality of an Inkjet-Grade Paper)

  • 이용규;이희명
    • 펄프종이기술
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    • 제37권2호
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    • pp.38-46
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    • 2005
  • Experimental work was carried out in order to produce a novel grade of ink-jet paper that has both high print-out quality and price competitiveness. Usually, silica and PVOH has been used for ink-jet paper to design the coating layer that has a hydrophilic and micro-porous structure. However, poor rheological characteristics and low productivity of the silica-PVOH system make the price of the ink-jet paper high. The main focus of this study was replacing the conventional silica (coating pigment) PVOH (binder) coating system with the new PCC (coating pigment) cationic starch (binder) coating system, and optimizing thecoating technology associated with PPC-cationic starch system. In this study, ink-jet print quality of PCC-coated papers was compared with that of silica-coated paper. Two types of PCC were used: conventional type and colloid type. It turned out that PCC C, a conventional coating pigment, has not given a desirable result: it showed high dot reproduction, but it gave low optical density. In spite of low dot reproduction, the qualities of PCC A were comparable or superior to those of silica in optical density, color reproduction, and the uniformity of printing surface. It was also shown that the problems that are happened when the dosage level of cationic starch was too low were varied with ink-type used in each printer. However, in the case of low binder level, the produced image was widely spread resulting fromtoo low optical density of images, or from the lack of bonding ability to set ink into coating surface.

Reproduction of Epstein-Barr Virus Infection and Pathogenesis in Humanized Mice

  • Fujiwara, Shigeyoshi
    • IMMUNE NETWORK
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    • 제14권1호
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    • pp.1-6
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    • 2014
  • Epstein-Barr virus (EBV) is etiologically associated with a variety of diseases including lymphoproliferative diseases, lymphomas, carcinomas, and autoimmune diseases. Humans are the only natural host of EBV and limited species of new-world monkeys can be infected with the virus in experimental conditions. Small animal models of EBV infection, required for evaluation of novel therapies and vaccines for EBV-associated diseases, have not been available. Recently the development of severely immunodeficient mouse strains enabled production of humanized mice in which human immune system components are reconstituted and express their normal functions. Humanized mice can serve as infection models for human-specific viruses such as EBV that target cells of the immune system. This review summarizes recent studies by the author's group addressing reproduction of EBV infection and pathogenesis in humanized mice.

TV 스피커의 저주파수 신호 재생 개선 (Improving Low Frequency Signal Reproduction in TV Audio)

  • 마나쉬 아로라;오윤학;김승훈;이혁재;장성철
    • 한국음향학회:학술대회논문집
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    • 한국음향학회 2004년도 춘계학술발표대회 논문집 제23권 1호
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    • pp.275-278
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    • 2004
  • In TV sound system, loudspeakers are subject to severe size constraints. The small size of the transducer affects the low frequency signal performance of the system. Bass signal performance contributes significantly to the user perceived sound quality and a good bass signal reproduction is essential. Increasing the sound energy in the bass signal range is an unviable solution since the gain required are exceedingly high and signal distortion occurs because of the speaker overload. Recently methods are being proposed to invoke low frequency illusion using psychoacoustic phenomena of the missing fundamental. This paper proposes a simple and effective signal processing method to create bass signal illusion in TV speakers using the missing fundamental effect, at a complexity of 12 MIPS on Motorola 56371 audio DSP.

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CMS를 사용한 별색 교정에 관한 연구 (A Study on Spot Color Proofing using ICC-based Color Management System)

  • 정청숙;강상훈
    • 한국인쇄학회:학술대회논문집
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    • 한국인쇄학회 2007년도 추계 학술논문 발표회
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    • pp.65-73
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    • 2007
  • The general commercial printing meets the customer's diverse demand by using spot color besides process four color. Especially, by using spot color for printing the enterprise's logo or specific color, we can see the effect of printing is getting better. The objective of this study was to examine the quality of spot color reproduction with Inkjet for 2 types of paper and Dye sublimation in ICC-based color management system. ICC profiles were generated for each device using ECI 2002 visual target and evaluated for the accuracy of each printer's color profile. The test chart consisting of Pantone color 1140 was selected to test the quality of spot color reproduction.

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