A DNA vaccine methodology using eukaryote expression vectors to produce immunizing proteins in the vaccinated hosts is a novel approach to the development of vaccine and immuno-therapeutics, and it has achieved considerable success over several infectious diseases and various cancers. To further enhance its efficiency, attempts were made to develop novel plasmid vectors containing multiple immunostimulatory CpG motifs, for rapid and strong immune response. First, a 2.9 kb compact plasmid vector (pVAC), containing CMV promoter, polycloning site, BGH poly(A) terminator, ampicillin resistance gene and pBR322 origin was constructed. A pVAC-hEPO was also constructed, which contained a human erythropoietin gene, for evaluating the transfection efficiency of naked plasmid DNA both in vitro and in vivo. To examine the adjuvant effect of multi-CpG motifs on naked plasmid DNA, 22 and 44 enriched and unmethylated CpG motifs were introduced into pVAC to generate pVAC-ISS1 and pVAC-ISS2, respectively. $100{\mu}g$ of pSecTagB, pVAC, pVAC-ISS1 or pVAC-ISS2 were each injected intramuscularly into the tibilias anterior muscle of Balb/c mice. The level of interleukin-6 induced in the mice injected with pVAC-ISS1 and pVAC-ISS2 were significantly elevated after 12 hours, which were almost 2 and 2.5 times higher than that in the mice injected with pSecTagB, respectively. These results suggest that DNA vaccine plasmids with enriched CpG motifs can induce rapid secretion of interleukin-6 by lymphocytes. In conclusion, these vectors can contribute to the development of adjuvant-free DNA vaccinations against infectious diseases and various cancers.
Kim, Su-Joong;Park, Won-Mok;Ryu, Ki-Hyun;Lee, Sang-Seon;Lee, Se-Yong
Korean Journal Plant Pathology
/
v.13
no.4
/
pp.248-254
/
1997
Expression vector (pGEX-Tu) for the coat protein (CP) gene of turnip mosaic virus Ca strain (TuMV-Ca) was constructed by incorporation of TuMV CP gene into pGEX-KG vector which had ${\beta}$-galactosidase gene and IPTG (isopropylthio-${\beta}$-D-galactoside) induction site. The results of ELISA and western hybridization indicated that optimal condition of the expression were when IPTG and western hybridization indicated that optimal condition of the expression were when IPTG induction was carried out on YTA medium with ampicillin in 2 hours after the E. coli seed inoculation ($A_{595}$=0.1/ml). TuMV CP gene was expressed with GST (Glutathion S-Transferase) gene fusion system, and the size of fusion protein was estimated to be 59kDa, for TuMV CP was 33 kDa and GST was 26 kDa.
ADFA, Morina;ROMAYASA, Ari;KUSNANDA, Arif Juliari;AVIDLYANDI, Avidlyandi;YUDHA S., Salprima;BANON, Charles;GUSTIAN, Irfan
Journal of the Korean Wood Science and Technology
/
v.48
no.1
/
pp.107-116
/
2020
Termiticidal and fungicidal activities of wood vinegar from Cinnamomum parthenoxylon (CP) stem wood have been evaluated against Coptotermes curvignathus and wood rotting fungi (Schizophyllum commune and Fomitopsis palustris). The utilized CP wood vinegar was produced in the operating temperature range 250-300℃ pyrolysis. A no-choice test was applied for evaluating termiticidal activity with 33 active termites and antifungal activity using the agar media assay. The result showed that an increase in the concentrations of CP wood vinegar significantly raised the mortality of termite. CP wood vinegar showed high termiticidal activity, organic acids (acetic acid 42.91%, 3-butenoic acid 6.89%, butanoic acid, 2-propenyl ester 2.26%), and ketones (1-hydroxy-2-propanone 5.14%, 3-methylcyclopentane-1,2-dione 2.34%) might be largely contributed to termiticidal activity in addition to other minor components. Furthermore, CP wood vinegar exhibited significant inhibition of fungal growth. These data showed that CP wood vinegar was more toxic to white-rot fungi (S. commune) than brown-rot (F. palustris). The results suggested that phenolic compounds from lignin degradation were responsible for good antifungal activity.
Xin Zhu;Kai Gao;Ziyi Zhang;Haiying Liu;Guiqin Yang
Animal Bioscience
/
v.36
no.11
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pp.1718-1726
/
2023
Objective: The objective of this study was to investigate the effect of supplementation with enzymolytic soybean meal (ESBM) on broilers fed low crude protein (CP) diets. Methods: A total of 360 one-day-old broilers were randomly assigned to six treatments with 6 replicates per treatment and 10 chicks per replicate for a period of 42 days. Chicks were fed a basal standard high-CP diet as a positive control (PC), a low-CP diet (reducing 10 g/kg CP from the PC) as a negative control (NC), or an NC + 0.5%, 1.0%, 1.5%, or 2.0% ESBM diet. Results: Compared to chicks fed the PC, chicks fed the NC had a decreased body weight gain (BWG, p<0.05) from 1 to 42 days, but supplementation with 2.0% ESBM restored BWG (p<0.05) and even linearly improved the feed conversion rate (FCR, p<0.05). Digestibility of CP and ether extract was increased (p<0.05) in chicks fed a 1.0% ESBM diet compared to the PC. With increasing levels of ESBM, nitrogen (N) excretion decreased (p<0.05). The addition of ESBM to the diet did not affect (p>0.05) serum concentrations of total protein, albumin and total cholesterol but led to a descending trend in triglycerides and an ascending trend in calcium and urea N at 42 days (p<0.10). There were no differences (p>0.05) in villus height (VH), crypt depth (CD), and VH/CD (V/C) of the duodenum and jejunum between the PC and NC at both 21 days and 42 days, while increasing dietary ESBM levels linearly (p<0.05) decreased CD and increased V/C of the duodenum and jejunum at both 21 days and 42 days. Conclusion: The findings indicated that ESBM could be used in broiler low-CP diets to improve production performance, decrease N excretion, and enhance intestinal health.
L-carnitine promotes mitochondrial ${\beta}$-oxidation of long chain fatty acids and their subsequent transport across the inner mitochondrial membrane. Although the role of L-carnitine in fatty acid metabolism has been extensively studied, its role in live performance and carcass responses of commercial broilers is less understood. The objective of this research was to determine if Lcarnitine fed at various levels in diets differing in CP and amino acids impacted on live performance and carcass characteristics of commercial broilers. Two floor pen experiments were conducted to assess the effect of dietary L-carnitine in grower diets. In Exp. 1, Ross${\times}$Hubbard Ultra Yield broilers were placed in 48 floor pens (12 birds/pen) and fed common diets to d 14. A two (0 or 50 ppm Lcarnitine) by three (173, 187, and 202 g/kg CP) factorial arrangement of treatments was employed from 15 to 35 d of age (8 replications/treatment). An interaction (p<0.05) in carcass yield indicated that increasing CP (187 g/kg) resulted in improved yield in the presence of L-carnitine. Increasing CP from 173 to 202 g/kg increased (p<0.05) BW gain and decreased (p<0.05) feed conversion and percentage abdominal fat. Feeding dietary L-carnitine increased back-half carcass yield which was attributable to an increase (p<0.05) in thigh, but not drumstick, yield relative to carcass. In Exp. 2, $Ross{\times}Ross$ 708 broilers were fed common diets until 29 d. From 30 to 42 d of age, birds were fed one of seven diets: i) 200 g/kg CP, 0 ppm L-carnitine; ii) 200 g/kg CP, 40 ppm L-carnitine; iii) 180 g/kg CP, 0 ppm L-carnitine; iv) 180 g/kg CP, 10 ppm L-carnitine; v) 180 g/kg CP, 20 ppm L-carnitine; vi) 180 g/kg CP, 30 ppm L-carnitine; and vii) 180 g/kg CP, 40 ppm L-carnitine (6 replications of 12 birds each). BW gain, feed conversion, mortality (30 to 42 d), and carcass traits (42 d) were measured on all birds by pen. There were no treatment differences (p<0.05). However, the addition of 40 ppm L-carnitine in the 200 g CP/kg diet increased (p = 0.06) thigh yields relative to BW in comparison to birds fed diets without L-carnitine, which was further confirmed via a contrast analysis (0 vs. 40 ppm L-carnitine in the 200 and 180 g CP/kg diets; p<0.05). These results indicated that dietary L-carnitine may heighten metabolism in dark meat of commercial broilers resulting in increased relative thigh tissue accretion without compromising breast accretion.
Barley has nutritional benefits due to its high dietary fiber content; therefore, the intake of whole barley grains is recommended. However, barley is often consumed in the fermented form because of the improved texture and digestibility. The present study was designed to elucidate the intracellular signaling pathway for macrophage activation by the polysaccharide BF-CP from fermented barley. BF-CP is a neutral polysaccharide, composed of neutral sugars, including glucose (70.7%), xylose (11.4%), and arabinose (9.0%). BF-CP exhibited macrophage-stimulatory activity by inducing the production of interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide in RAW 264.7 macrophages. Further, BF-CP treatment strongly increased the IL-6 and $TNF-{\alpha}$ gene expression in a concentration-dependent manner. Signal transduction experiments using immunoblotting showed that BF-CP phosphorylated mitogen-activated protein kinases (MAPKs), such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38, and nuclear factor $(NF)-{\kappa}B$, in RAW 264.7 cells in a concentration-dependent manner. These results suggest that BF-CP activates the macrophages via MAPK and $NF-{\kappa}B$ pathways, and also induces an increase in the production of cytokines.
Kim Yong-Gyun;Basio Neil A.;Ibrahim Ahmed M.A.;Bae Sung-Woo
Korean journal of applied entomology
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v.45
no.1
s.142
/
pp.15-24
/
2006
Inhibitor kB (IkB)-like gene has been found in the genome of Cotesia plutellae bracovirus (CpBV), which is the obligatory symbiont of an endoparsitoid wasp, C. plutellae. The open reading frame of CpBV-IkB was 417 bp and encoded 138 amino acids. Four ankyrin repeat domains were found in CpBV-IkB, which shared high homology with other known polydnavirus IkBs. Considering a presumptive cellular IkB based on Drosophila Cactus, CpBV-IkB exhibited a truncated structure with deletion of signal-receiving domains, which suggested its irreversible inhibitory role in NFkB signal transduction pathway of the parasitized host in response to the wasp parasitization. CpBV-IkB was expressed only in the parasitized diamondback moth, Plutella flostella. Its expression was estimated by quantitative RT-PCR during parasitization period, showing a constitutive expression pattern from the first day of parasitization. An indirect functional analysis of CpBV-IkB was conducted and suggested a hypothesis of host antivirus inhibition.
Fang, Lin Hu;Jin, Ying Hai;Do, Sung Ho;Hong, Jin Su;Kim, Byung Ock;Han, Tae Hee;Kim, Yoo Yong
Asian-Australasian Journal of Animal Sciences
/
v.32
no.4
/
pp.556-563
/
2019
Objective: This experiment was conducted to investigate the effect of reducing dietary metabolic energy (ME) and crude protein (CP) levels on growth performance, blood profiles, and nutrient digestibility in weaning pigs. Methods: A total of 240 crossbred pigs ($Duroc{\times}[Landrace{\times}Yorkshire]$) with an average body weight of $8.67{\pm}1.13kg$ were used for a 6-week feeding trial. Experimental pigs were allotted to a $2{\times}3$ factorial arrangement using a randomized complete block design. The first factor was two levels of dietary ME density (low ME level, 13.40 MJ/kg or high ME level, 13.82 MJ/kg) and the second factor was three dietary CP levels based on subdivision of early and late weaning phases (low CP level, 19.7%/16.9%; middle CP level, 21.7%/18.9%; or high CP level, 23.7%/20.9%). Results: Over the entire experimental period, there were no significant difference in body weight among groups, but a decrease in diet energy level was associated with an increase in average daily feed intake (p = 0.02) and decrease in gain-feed ratio (G:F) ratio (p<0.01). Decreased CP levels in the diet were associated with a linear increase in average daily gain (p<0.05) and quadratic increase in G:F ratio (p<0.05). In the early weaning period, blood urea nitrogen concentration tended to increase when ME in the diet decreased and decrease when CP level in the diet decreased (p = 0.09, p<0.01, respectively). Total protein concentration tended to increase when CP level was reduced (p = 0.08). In the late weaning period, blood urea nitrogen concentration decreased linearly as CP level decreased (p<0.01). The CP and crude fat digestibility decreased when ME was decreased by 0.42 MJ/kg (p = 0.05, p = 0.01, respectively). The CP digestibility increased linearly as CP level decreased (p = 0.01). Conclusion: A weaning pig diet containing high ME level (13.82 MJ/kg) and low CP level (19.7%/16.9%) can improve pig growth performance and nutrient digestibility.
Purpose: Tissue inhomogeneity such as lung affects tumor dose as well as transmission dose in new concept of on-line dosimetry which estimates tumor dose from transmission dose using the new algorithm. This study was carried out to confirm accuracy of correction by tissue density in tumor dose estimation utilizing transmission dose. Methods: Cork phantom (CP, density $0.202\;gm/cm^3$) having similar density with lung parenchyme and polystyrene phantom (PP, density $1.040\;gm/cm^3$) having similar density with soft tissue were used. Dose measurement was carried out under condition simulating human chest. On simulating AP-PA irradiation, PPs with 3 cm thickness were placed above and below CP, which had thickness of 5, 10, and 20 cm. On simulating lateral irradiation, 6 cm thickness of PP was placed between two 10 cm thickness CPs additional 3 cm thick PP was placed to both lateral sides. 4, 6, and 10 MV x-ray were used. Field size was in the range of $3{\times}3$ cm through $20{\times}20$ cm, and phantom-chamber distance (PCD) was 10 to 50 cm. Above result was compared with another sets of data with equivalent thickness of PP which was corrected by density. Result: When transmission dose of PP was compared with equivalent thickness of CP which was corrected with density, the average error was 0.18 (${\pm}0.27$) % for 4 MV, 0.10 (${\pm}0.43$) % for 6 MV, and 0.33 (${\pm}0.30$) % for 10 MV with CP having thickness of 5 cm. When CP was 10 cm thick, the error was 0.23 (${\pm}0.73$) %, 0.05 (${\pm}0.57$) %, and 0.04 (${\pm}0.40$) %, while for 20 cm, error was 0.55 (${\pm}0.36$) %, 0.34 (${\pm}0.27$) %, and 0.34 (${\pm}0.18$) % for corresponding energy. With lateral irradiation model, difference was 1.15 (${\pm}1.86$) %, 0.90 (${\pm}1.43$) %, and 0.86 (${\pm}1.01$) % for corresponding energy. Relatively large difference was found in case of PCD having value of 10 cm. Omitting PCD with 10 cm, the difference was reduced to 0.47 (${\pm}$1.17) %, 0.42 (${\pm}$0.96) %, and 0.55 (${\pm}$0.77) % for corresponding energy. Conclusion When tissue inhomogeneity such as lung is in tract of x-ray beam, tumor dose could be calculated from transmission dose after correction utilizing tissue density.
Junyoung Mun;Habeeb Tajudeen;Abdolreza Hosseindoust;Sanghun Ha;Serin Park;Jinsoo Kim
Journal of Animal Science and Technology
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v.66
no.3
/
pp.493-503
/
2024
The aim of this experiment was to evaluate the effects of low crude protein (CP) level with essential amino acids (AA) addition on growth performance, nutrient digestibility, microbiota, and volatile fatty acid composition in growing pigs. A total of 160 growing pigs (Landrace × Yorkshire × Duroc [LYD]; average initial body weight 16.68 ± 0.12 kg) were randomly allotted to one of the four treatments on the basis of initial body weight. A randomized complete block design was used to conduct this experiment in the Research Center of Animal Life Sciences at Kangwon National University. There were ten pigs/replicate with four replicates in each treatment. The treatments include; CON (Control, 17.2% dietary CP level), low protein (LP)-1.10 (15.7% dietary CP level + 1.10% lysine level), LP-1.15 (15.7% dietary CP level + 1.15% lysine level), LP1.2 (15.7% dietary CP level + 1.20% lysine level). The pigs fed CON and LP-1.2 diet showed greater final body weight than that of LP-1.1 diet (p < 0.05). Although average daily gain, average daily feed intake, and feed efficiency did not show any difference in phase 2 and 3, average daily gain and feed efficiency was significantly greater in CON and LP-1.20 in phase 1. However, the average daily feed intake did not show any difference during the experimental period. Isobutyric acid and isovaleric acid composition of LP treatments were lower than CON treatment in phase 2. Total branched chain fatty acid composition was significantly lower in LP treatment in phases 1 and 2. However, there was no significant difference among treatments in phase 3. The results of this study underscore the importance of AA supplementation when implementing a low-protein diet during the early growth phase (16-50 kg) in pigs.
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