Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.
The objective of this experiment was to select the best strain of lactic acid bacteria for the manufacture of new type of yogurt with rice powders. Changes in pH, titratable acidity, viable cell counts, viscosity, organic acid contents, carbohydrates during fennentation were monitored and sensory evaluation were examined. The yogurt added with 4% rice or skim milk powders and L. salivarius ssp. salivarius culture did not reach pH 4.5, because the production of acids in this media for the culture was weak. The yogurt added with 4% rice or skim milk powder with L. casei, the pH was low and the titratable acidity was high, and therefore the quality of yogurt after 8 hours from fermentation was not high. The yogurt added with 4% rice or skim milk powders with a mixed culture of B. longum, L. acidophilus, Streptococcus salivarius ssp. thermophilus was considered best for achieving pH 4.5 and titratable acidity of 1.0 % from 8 to 14 hours. The yogurt with a mixed culture had more acetic acid. Galactose was accumulated when L. salivarius ssp. salivarius or the mixed culture were used for fermenting yogurt. In sensory evaluation, the yogurt with the mixed culture received high overall sensory score. From these results, a mixed culture of B. longum, L. acidophilus, Streptococcus salivarius ssp. thermophilus was identified as the best for the manufacture of yogurt added with rice powder.
Kang, Yang-Soon;Kim, Wan Joong;Hwang, Duck Sang;Kim, Hee Kyu
KOREAN JOURNAL OF CROP SCIENCE
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v.65
no.1
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pp.30-39
/
2020
A new silicate coating technology was developed which reduces the impact of dust and loosening during seeding compared to existing silicate-coatings (Seed/Si/Zeolite), and therefore can lower the production costs of rice cultivation. In this method, 100 g of rice seed is coated with 18 mL of liquid silicic acid and then dressed with a mixture containing 80 g of dolomite and 5 g of iron. To determine the most effective method of application and ensure that seedlings developed healthily, a series of experiments were carried out. Infected seeds scattered in seedling boxes and pots (soil and hydroponic) were coated dry, without disinfection. In comparison to the seed which were not treated with the silicate-coating, the new seed (A) were 1.84 times heavier in weight, and were also improved in terms of coating strength and coating color. Compared to the seedlings grown from the non-coated seed, those grown from the new silicate-coated seed were of significantly higher quality (weight/length) and had erect, dark greenish leaves, which are ideal plant characteristics. This was most likely due to increased silicate uptake. The symptoms of bakanae disease in the non-coated seed peaked after 38 days to 54.2%, whereas the control value was 68.8% in the new silicate-coated seed (A). In the infected seedlings grown from the new silicate-coated rice seed, subnormal macro-conidia, namely, a sickle shape spore without a septum; a straight oblong shape spore without a septum and with a thick cell wall; and inter-septal necrosis of a normal spore were detected. It is believed that the strong alkalinity of silicic acid have acted as unfavorable conditions for pathogenicity. In seedlings grown from the new silicate coated rice seed under hydroponic conditions without nutrients, normal root activity and growth was maintained without leaf senescence. Therefore, it was possible to reduce the rate of fertilization. In the future, a new silicate-coated rice seed was required for the study of minimal nutrition for anti-aging of seedlings.
The identity of toxin producers remains only hypothesis unless there were identified by strain isolation and analytical confirmation of both the cyanotoxin production and the genetic identity of the monoculture. The purposes of this study were to identify a morphologic and phylogenetic classification in Aphanizomenon flos-aquae strains isolated from the Nakdong River and to investigate the potential ability of the strains to produce toxins such as saxitoxin and cylindrospermopsin using target genes. The 16S rRNA and sxtA, sxtI, cyrA, cyrJ genes were analyzed on two strains (DGUC001, DGUC003) isolated from the Nakdong River. Morphological features of the strains were observed a shape of aggregated trichomes in parallel fascicles which can reach up to macroscopic size and a hyaline terminal cell without aerotope. In addition, the 16S rRNA phylogenetic analyses showed that the strains were identified as the same species with high genetic similarity of 98.4% and grouped within a monospecific andsupported cluster I of Aphanizomenon flos-aquae selected from GenBank of the NCBI. The cyrA and cyrJ genes encoding for the cylindrospermopsin-biosynthesis were not detected in the present study. The sxtA gene was in detected both the two strains, whereas the sxtI gene which had been suggested as a suitable molecular marker to detect saxitoxin-producing cyanobacteria was not found both the strains. Thus, the two strains isolated from Nakdong River were identified as the same species of Aphanizomenon flos-aquae Ralfs ex Bornet et Flahault 1888, the two strains were confirmed as potential non-producing strains of the saxitoxin and cylindrospermopsin.
Dendritic cells (DCs) are the only antigen presenting cells (APCs) capable of initiating immune responses, which is crucial for priming the specific cytotoxic T lymphocyte (CTL) response and tumor immunity. Upon activation by DCs, CD4+ helper T cells can cross-prime CD8+ CTLs via IL-12. However, recently activated DCs were described to prime in vitro strong T helper cell type 1 $(Th_1)$ responses, whereas at later time points, they preferentially prime $Th_2$ cells. Therfore, we examined in this study the optimum kinetic state of DCs activation impacted on in vivo priming of tumor-specific CTLs by using ovalbumin (OVA) tumor antigen model. Bone-marrow-derived DCs showed an appropriate expression of surface MHC and costimulatory molecules after 6 or 7-day differentiation. The 6-day differentiated DCs pulsed with OVA antigen for 8 h (8-h DC) and followed by restimulation with LPS for 24 h maintained high interleukin (IL)-12 production potential, accompanying the decreased level in their secretion by delayed re-exposure time to LPS. Furthermore, immunization with 8-h DC induced higher intracellular $interferon(IFN)-{\gamma}+/CD8+T$ cells and elicited more powerful cytotoxicity of splenocytes to EG7 cells, a clone of EL4 cells transfected with an OVA cDNA, than immunization with 24-h DC. In the animal study for the evaluation of therapeutic or protective antitumor immunity, immunization with 8-h DC induced an effective antitumor immunity against tumor of EG7 cells and completely protected mice from tumor formation and prolonged survival, respectively. The most commonly used and clinically applied DC-based vaccine is based on in vitro antigen loading for 24 h. However, our data indicated that antigen stimulation over 8 h decreased antitumor immunity with functional exhaustion of DCs, and that the 8-h DC would be an optimum activation state impacted on in vivo priming of tumor-specific CTLs and subsequently lead to induction of strong antitumor immunity.
Kim, Eun-Ju;Song, Bit-Na;Jeong, Da-Som;Kim, So-Young;Cho, Yong-Sik;Park, Shin-Young
Journal of Life Science
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v.27
no.11
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pp.1315-1323
/
2017
Turmeric is a rhizomatous herbaceous perennial plant (Curcuma longa (CL)) of the ginger family, Zingiberaceae. A yellow-pigmented fraction isolated from the rhizomes of CL contains curcuminoids belonging to the dicinnamoyl methane group. Curcumin is an important active ingredient responsible for the biological activity of CL. However, CL is not usually used as a food source due to its bitter taste. The present study was designed to determine the effect of the CL fermented by Rhizopus oryzae (FCL) on pro-inflammatory factors such as nuclear factor ${\kappa}B$ ($NF-{\kappa}B$), tumor necrosis factor alpha ($TNF-{\alpha}$), interleukin-6 (IL-6), nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-induced RAW 264.7 cell line. The cell viability was determined by MTT assay. To evaluate the anti-inflammatory effect of FCL 80% EtOH extracts, IL-6 and $TNF-{\alpha}$ were measured by ELISA kit. Also, the amount of $NO/PGE_2/NF-{\kappa}B$ was measured using the $NO/PGE_2/NF-{\kappa}B$ detection kit and the iNOS/COX-2 expression was measured by Western blotting. The results showed that the FCL reduced NO, $PGE_2$, iNOS, COX-2, $NF-{\kappa}B$, IL-6 and $TNF-{\alpha}$ production without cytotoxicity. These results suggest that FCL extracts may be a developed the functional food related to anti-inflammation due to the significant effects on inflammatory factors.
Acute respiratory distress syndrome (ARDS) is a kind of acute lung injury characterized by inflammatory disruption of alveolar-capillary barrier and notorious for its high mortality. Neutrophils cause cell damage through the production of free radicals, inflammatory mediators, and proteases in ARDS. $PLA_2$ might serve a primary regulatory role in the activation of neutrophils. This present study was performed to elucidate the effect of rutin known as $PLA_2$ inhibitor on ARDS induced by endotoxin. Endotoxin had increased lung myeloperoxidase (MPO) activity, BAL (bronchoalveolar lavage) protein content, numbers of neutrophils in BALF (bronchoalveolar lavage fluid) compared with those of control rat (p<0.001). In addition, histological evidence of lung injury was correlated with neutrophil influx into alveolar space and cerrous perhydroxide granules were found in lining of endothelial cell, alveolar type I, II cells. In contrast, pretreated group of rutin had significantly decreased all of the parameters (p<0.001). These data suggest that inhibition of $PLA_2$ is one step approach that block the process of ARDS. Accordingly, we conclude that rutin can be used as the prophylactic agent for ARDS on the bases of these experimental results.
Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
Radiation Oncology Journal
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v.22
no.2
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pp.145-154
/
2004
Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.
This study was conducted to select some useful plants as functional material candidates. A total of 38 plants were preliminarily screened for the anti-inflammatory and antioxidant activities. The preliminarily selected 8 plants were further investigated to verify the in vitro inhibitory effect on inflammation and oxidative stress. Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) inhibited the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-treated RAW 264.7 cells. Eupatorium japonicum (leaf) suppressed the expression of cyclooxygenase-2 (COX-2), whereas Boehmeria platanifolia (root) and Prunus yedoensis (branch) inhibited the transcription of nuclear factor-kappa B (NF-${\kappa}B$). Treatment with the extracts ($2.5{\sim}20{\mu}g/ml$) of Abutilon theophrasti (leaf, flower/seed) and Hemistepta lyrata (stem) did not show toxicity on RAW 264.7 cell proliferation, but treatment with $2.5{\mu}g/ml$ of Boehmeria platanifolia (root) exhibited cell toxicity. Carpinus coreana (branch) and Prunus yedoensis (branch) showed potent scavenging activities on peroxynitrite. Akebia quinata (flower), Carpinus coreana (branch), and Prunus yedoensis (branch) effectively inhibited reactive oxygen species (ROS). Abutilon theophrasti (leaf), Boehmeria platanifolia (root), Carpinus coreana (branch), and Eupatorium japonicum (leaf) exhibited strong inhibitory capacity with regard to nitric oxide (NO) production. The results suggested that Abutilon theophrasti (leaf) has in vitro anti-inflammatory and antioxidant activities, and that is a useful functional material candidate.
Potatoes were first introduced outside the Andes region four centuries ago, and have become an integral part of much of the world's food. Potatoes were first introduced into Europe in the 16th century and Korea in the early 19th century. In the nutritional aspects, potatoes contain abundant vitamins and minerals, as well as an assortment of phytochemicals such as carotenoids and natural phenols. Chlorogenic acid constitutes up to 90% of potato natural phenols. Due to the high content of potato functional compounds, it has known that potatoes are effective in the prevention of various human diseases. Recently, color-fleshed potatoes 'Hongyoung' and 'Jayoung' were developed by RDA, and it has reported that they have high content of anthocyanin. Additionally they show higher radical scavenging activity compared to white or yellow fleshed potatoes. So it will be expected that the consumption of color-fleshed potatoes grandually increase by pre-peeled potatoes and color potato chips. This study was conducted to enhance the peel of color-fleshed potatoes utilization and to determine the biological activity of peel of color-fleshed potatoes extract. The anti-inflammatory effects on ethanol extract and its solvent fraction were also evaluated. The anti-inflammatory activities of $CHCl_3$ fraction was evaluated for inhibitory activities against lipopolysacchride(LPS) induced nitric oxide(NO) and prostaglandin $E_2(PGE_2)$ production as well as inducible nitric oxide synthase(iNOS) and cyclo oxygenase-2(COX-2) protein expressions in RAW264.7 cell lines. The fraction inhibitory activity for both tests with $IC_{50}$ values showed in the ranges of $25{\sim}50{\mu}g/ml$. This result revealed that $CHCl_3$ fraction of Jayoung's peel is expected to be good candidate for development into source of anti-inflammatory agent.
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