This study was conducted to research the morphological characteristics and analyze the genetic diversity by using RAPD in Calanthe species native to Korea. Nine samples were selected by flower color and 19 morphological characteristics. In the length and width of leaf, dorsal sepal, the lateral sepal, the petal, the central lip, and the lateral lip, C. discolor was the shortest and narrowest, respectively, but C. sieboldii was the longest and the widest, respectively. The flower stalk length was the shortest in C. discolor, and the longest in C. sieboldii. Three variants were the intermediate between C. discolor and C. sieboldii in the above morphological characteristics, but spur length was the longest in C. discolor, the shortest in C. sieboldii, and intermediate in the variants. The ovary length in C. discolor was shortest and C. sieboldii and variants were similar with each other. The flower color of C. discolor were brownish red, the value of CIE Lab was between 40 and 50. The flower color of C. sieboldii was yellowish, the value of CIE Lab was between 110 and 130. And variants had various colors between 50 to 70 in the value of CIE Lab. By analyzing multiple band patterns of PCR products, 154 bands were selected as polymorphic RAPD markers. The analysis of genetic similarity of Calanther species using RAPD showed that C. discolor and C. sieboldii are more distant from each other than variants, and these results demonstrated that genetic position of variants located between C. discolor and C. sieboldii.
High-temperature requirement A2(HtrA2) has been known as a human homologue of bacterial HtrA that has a molecular chaperone function. HtrA2 is mitochondrial serine protease that plays a significant role in regulating the apoptosis; however, the physiological function of HtrA2 still remains elusive. To establish experimental system for the investigation of new insights into the function of HtrA2 in mammalian cells, we first obtained $HtrA2^{+/+}$ and $HtrA2^{-/-}$ MEF cells lines and identified those cells based on the expression pattern and subcellular localization of HtrA2, using immunoblot and biochemical assays. Additionally, we observed that the morphological characteristics of $HtrA2^{-/-}$ MEF cells are different form those of $HtrA2^{+/+}$ MEF cells, showing a rounded shape instead of a typical fibroblast-like shape. Growth rate of $HtrA2^{-/-}$ MEF cells was also 1.4-fold higher than that of $HtrA2^{+/+}$ MEF cells at 36 hours. Furthermore, we verified both MEF cell lines induced caspsase-dependent cell death in response to apoptotic stimuli such as heat shock, staurosporine, and rotenone. The relationship between HtrA2 and heat shock-induced cell death is the first demonstration of the research field of HtrA2. Our study suggests that those MEF cell lines are suitable reagents to further investigate the molecular mechanism by which HtrA2 regulates the balance between cell death and survival.
The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.
Kwak, Chung Shil;Kim, Mi-Ju;Kim, Sun Gi;Park, Sunyeong;Kim, In Gyu;Kang, Heun Soo
Journal of Nutrition and Health
/
v.52
no.6
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pp.529-539
/
2019
Purpose: Sprouts of evening primrose (Oenothera laciniata, OL) were reported to have high contents of flavonoids and potent antioxidant activity. This study examined the antioxidant and antiobesity activities of OL sprouts to determine if they could be a natural health-beneficial resource preventing obesity and oxidative stress. Methods: OL sprouts were extracted with 50% ethanol, evaporated, and lyophilized (OLE). The in vitro antioxidant activity of OLE was examined using four different tests. The antiobesity activity and in vivo antioxidant activity from OLE consumption were examined using high fat diet-induced obese (DIO) C57BL/6 mice. Results: The IC50 for the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging and superoxide dismutase (SOD)-like activities of OLE were 26.2 ㎍/mL and 327.6 ㎍/mL, respectively. OLE exhibited the ferric reducing antioxidant power (FRAP) activity of 56.7 ㎍ ascorbic acid eq./mL at 100 ㎍/mL, and an increased glutathione level by 65.1% at 200 ㎍/mL compared to the control in the hUC-MSC stem cells. In an animal study, oral treatment with 50 mg or 100 mg of OLE/kg body weight for 14 weeks reduced the body weight gain, visceral fat content, fat cell size, blood leptin, and triglyceride levels, as well as the atherogenic index compared to the high fat diet control group (HFC) (p < 0.05). The blood malondialdehyde (MDA) level and the catalase and SOD-1 activities in adipose tissue were reduced significantly by the OLE treatment compared to HFC as well (p < 0.05). In epididymal adipose tissue, the OLE treatment reduced the mRNA expression of leptin, PPAR-γ and FAS significantly (p < 0.05) compared to HFC while it increased adiponectin expression (p < 0.05). Conclusion: OLE consumption has potent antioxidant and antiobesity activities via the suppression of oxidative stress and lipogenesis in DIO mice. Therefore, OLE could be a good candidate as a natural resource to develop functional food products that prevent obesity and oxidative stress.
The objective of the study was to observe the effect of n-3 PUFA on cell proliferation and apoptosis by determining mRNA and protein of COX-2 and eicosanoid product and the mRNA and protein of Bu and Bcl-2 related to apoptosis in colon carcinogenesis of 1,2- dimethylhydrazine (DMH)-treated rats. Ninety male Sprague Dawley rats weighing about 170g were divided into 3 groups, control and n-3 PUFA supplemented groups (FO group: 6.2 mmoles n-3 PUFA; 2FO group: 12.4 mmoles n-3 PUFA) and fed experimental diet for 14 weeks. All rats were intramuscularly injected with DMH 15 mg/kg twice a week for 6 weeks to deliver total dose of 180 mg/kg body weight. Compared with the control group, 6.2 mmoles n-3 PUFA significantly reduced the levels of mRNA and protein expression of COX-2 and 2-series eicosanoids ($TXB_{2}$ and $PGE_{2}$) and decreased cell proliferation in colonic mucosa. However, high levels of n-3 PUFA supplementation significantly increased the levels of mRNA and protein expression of COX-2, TXB2 and PGE2. and increased cell proliferation which was similar level to that of control group. Compared with the control group, n-3 PUFA, regardless of the amount, significantly increased apoptotic index in colonic mucosa. Western blot and RT-PCR analyses showed that the levels of mRNA and protein expression of Bax were significantly increased by 6.2 mmoles n-3 PUFA, but decreased by 12.4 mmoles n-3 PUFA. The analyses also showed the levels of mRNA and protein expression of Bcl-2 were significantly reduced by 6.2 mmoles n-3 PUFA, but increased by 12.4 mmoles n-3 PUFA. The ratio of Bcl-2/Bax in mRNA and protein was significantly reduced by 6.2 mmoles n-3 PUFA but increased by 12.4 mmoles n-3 PUFA. Overall, these results indicate that n-3 PUFA could be effective in preventing colon carcinogenesis by reducing cell proliferation with lower level of COX-2 and 2-series eicosanoid, and increasing apoptosis by inducing pro-apoptotic gene, Bax and inhibiting anti-apoptotic gene, Bcl-2 in the colonic mucosa of DMH-treated rats. However, high level of n-3 PUFA supplementation could stimulate colon carcinogenesis by increasing cell proliferation and inhibiting apoptosis. (Korean J Nutrition 38(10): 807$\sim$816,2005)
The DNA sequencer is known to be more sensitive for the quality of template DNA, method of purification followed by sequencing reaction, and gel concentration. Therefore, we investigated optimal conditions for template preparation, purification, sequencing reaction, gel concentration, and injection medium. For plasmid prepara- tion, using chloroform instead of phenol improved the average read length from 532 bp to 684 bp. The addition of 2.5% DMSO sequencing PCR reaction resulted in 200 bp longer sequences. Purification using 50 mM EDTA and 0.6 M Sodium acetate(pH 8.0) presented 20 bp longer sequences than that using 50 mM EDTA(pH 8.0) and 0.6 M sodium acetate(pH 5.2). The injection for sequencing analysis using ABI formamide presented 90 bp longer sequences than that of using formamide deionized by resin. Moreover, there were 150 bp more readable sequences in 3.6% PAGE gel than in 4%. Consequently, it was concluded that an average of 700 bp per reaction with 85% accuracy can be obtained by the following optimal conditions: template preparation using chloroform, 2.5% DMSO, 50 mM EDTA and 0.6 M sodium acetate(pH 8.0), ABI formamide and 3.6% gel concentration.
This study was conducted to develop a set of EST-SSR marker for the purity test of commercial F1 hybrid cultivars in the watermelon. A total of 353 EST-SSR were selected and tested on seven F1 cultivars and their 11 parental lines achieved from NH Seeds Inc., Korea. Among tested 96 primer sets, WMU0056 for 'Orange', WMU0400 for 'Heukbo', WMU0056 and WMU0400 for 'Sindong', and WMU0056 and WMU0400 for 'Serona' revealed polymorphisms between the parental lines and heterozygosity from these F1 cultivers. Of 122 primer sets tested for 'Haedong', WMU0056, WMU0400, WMU0580, WMU1211, WMU4136, and WMU448 showed polymorphisms that were appropriate for the F1 purity test. WMU0056 and WMU0400 can be useful for 'Haedong', as well. Relatively low polymorphisms between parental lines were detected for 'Kulnara'(5%) and 'Hwangpea'(2%), and therefore, all 353 primer sets were tested on these cultivars. As the result, WMU5339 and WMU7003 were found to be useful for the F1 purity test in 'Kulnara' and 'Hwangpea', respectively. Using these EST-SSR markers developed by ICuGI, hybridity of the seeds for four F1 cultivars produced from farmers was evaluated, and levels of the F1 purity higher than 97.5% was observed from all seed populations. Our results indicated that the watermelon EST-SSR marker information posted in ICuGI could be utilized for developing codomiant and locus-specific markers that are highly effective for the F1 purity test.
Kim, Dong In;Kim, Hyun Jung;Yun, Jong Moon;Lee, Ji Hye;Han, So Jung;Kim, Ha Eun;Jang, Min Jung;An, Bong Jeun
Food Science and Preservation
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v.25
no.1
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pp.107-116
/
2018
The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-$1{\beta}$ (IL-$1{\beta}$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and prostaglandin $E_2$ ($PGE_2$) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and $ABTS^+$ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were $217.04{\pm}2.98$, $156.72{\pm}3.90$, and $182.88{\pm}3.02mg\;TAE/g$, respectively, while the electron donating abilities at $1,000{\mu}g/mL$ of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The $ABTS^+$ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at $50{\mu}g/mL$ of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.
In this study, monokaryons of "Heukari" (Pleurotus ostreatus) and "Hosan" (Pleurotus pulmonarius) were separated to remove the cell wall, and a cross-species protoplast fusion was developed through chemical treatment with polyethylene glycol. The protoplast-fused PF160306 and PF160313 strains have a culture period of 10 and 2 days shorter than that of the "Heuktari" and "Hosan" cultivars, respectively. Furthermore, the growth of the strains was faster than that of the existing cultivars. The yield was 135.9 g per bottle, which was approximately 8% higher than that of the commercially available "Hosan" cultivar; however, it was not statistically significant. A growth survey was conducted after treatment at five temperatures (15, 18, 21, 23, and 25℃). The growth of the strains accelerated with the increase in temperature. However, at 21℃, the yellow color of pileus was the brightest. Band pattern, assessed using URP Primer 7, was similar to that of the "Hosan" cultivar. The DPPH radical scavenging capacity and polyphenol content were 62.5% and 43.5 mg/mL, respectively, for "Sunjung" and 65.7% and 49.9 mg/mL, respectively, for PF160313. Furthermore, the antihypertensive activities of the "Sunjung" cultivar and PF160313 were similarly high at 74% and 75%, respectively. In conclusion, cross-species hybridization via the protoplast fusion technique can be used for obtaining primary data for mushroom breeding to develop new varieties. In addition, the protoplast fusion technique might aid in expanding the market for yellow mushrooms.
Kim, Chulyoung;Choi, Duyeol;Kang, Jeong Hun;Ahmed, Shabbir;Kil, Eui-Joon;Kwon, Gimyeon;Lee, Gwan-Seok;Kim, Yonggyun
Korean journal of applied entomology
/
v.60
no.4
/
pp.387-401
/
2021
Thrips infesting hot peppers were monitored in greenhouses using yellow sticky traps. In addition, the hot peppers infected with tomato spotted wilt virus (TSWV) were observed during the monitoring period. The flower thrips (Frankliniella intonsa) were initially trapped at a low density just after transplanting seedlings of hot peppers at late March. The western flower thrips (Frankliniella occidentalis) were trapped after mid April. These two thrips represented more than 98% of the total thrips attracted to the traps after May, in which F. intonsa showed higher occurrence frequency than F. occidentalis. The total number of thrips had two peaks at mid May with a small and short-term peak and at June-July with a large and long-term peak. The trapped thrips exhibited inconsistent sex ratios, suggesting a seasonal parthenogenesis. Different geographical populations were varied in cytochrome oxidase I sequences, in which local populations in Andong shared a high sequence similarity. TSWV-infected hot peppers, which might be mediated by these two thrips species, were observed and confirmed by an immunoassay kit and a molecular diagnosis using RT-PCR. In addition, the TSWV was detected in F. occidentalis collected from the infected hot peppers. Three open reading frames (NSS, N, and NSM) of the isolated TSWV genomes were sequenced and showed multiple point mutations containing missense mutations among geographical variants. When the isolated TSWV was fed to nonvirulent thrips of F. occidentalis, the virus was detected in both larvae and adults. However, the viral replication occurred in larvae, but not in adults.
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