• 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.

• Journal of fish pathology
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• 제10권1호
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• pp.1-14
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• 1997

Major object of this study was to investigate the causative organism of th e diseased cultured fingerling of sea bass, L japonicus. The experimental results are summarized as follows ; Staphylococcus epidermidis, isolated from the liver, kidney, spleen and brain, was considered to be the causative organism. External symptoms of this disease were congestion and hemorrhages in eyes. Anatomical symptoms were hemorrhage of brain, congestion of liver, and slight swelling of kidney and spleen. Growth of the isolates was good on BHIA, HIA and Staphylococcus No. 110. The growth occurred at a range(optimum) of $10\sim45^{\circ}C(35\sim40^{\circ}C)$, 0~9% (1~3%) of NaCl concentration and pH 4~10(8). DNase and coagulase production of all isolated strains were nagative, but was positive in hemolysis. Urease was positive reaction, and novobiocin resistance was nagative. Acid was produced anaerobically from glucose and maltose. Acid was produced aerobically from glucose, galactose, sucrose, maltose and dextrine. But gas was not produced from any carbohydrates. When the isolated strain was injected intramuscularly on fingerling of sea bass, L japonicus, it had virulence at $1.7{\times}10^{10}$ viable cells/$m\ell$ for all fish examined but no virulence at $1.7{\times}10^4$ viable cells/$m\ell$. Bacitracin, Erythromycin and Nofloxacin were observed as bacteriostatic agents to the strain, but Colistin, Gentamicin and Nalidixic acid were not. There were remarkable congestion of the brain, regressive necrosis of the liver, and showed necrosis of the epithelial cells of renal tubules in kidney tissues.

• Korean journal of applied entomology
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• 제10권2호
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• pp.103-107
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• 1971

In this report, an experiment has been conducted to test the quantitative changes of nucleic acids in the nuclei of the epidermal cell of the galls, caused by Eriophyes kuko Kishida on the leaf of Lycium chinense Mill by means o( microspectrophotometric techniques, the two-wave-length methods. And the sizes of the epidermal cells and nuclei have been measured. The experimental results were summarized as follows: 1) It has been found that as the gall grows, the diameter of the epidermal cells and their nuclei increased and they were larger than those of tile healthy ones. 2) Microspectrophotometric measurement of nuclei by the 'Two-wave-length method' after staining with Feulgen reagent showed no changes in DNA content in the early stage of the gall. As the gall matured, however, DNA content of the gall increased more than that of the healthy leaf. 3) RNA-measurement of nuclei stained with Azur-B in DNase treated epidermal cells of the gall revealed that temporary increase in RNA content occurred in early to middle stages after the gall formation. As the gall matured, however, RNA content of the gall decreased more as against that of the healthy leaf.