• Title/Summary/Keyword: E2 enzyme

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Production and Characterization of Monoclonal Antibodies Specific to PAT Protein Expressed in Genetically Modified Herbicide-Resistance Maize (제초제 내성 유전자 변형 옥수수 중 PAT단백질에 특이한 단크론성 항체의 생산과 특성 확인)

  • Kim, Sol-A;Lee, Jeong-Eun;Shim, Won-Bo;Kang, Sung-Jo;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.33 no.3
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    • pp.193-199
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    • 2018
  • In this study, PAT protein of genetically modified maize was prepared from the recombinant E. coli strain BL21 (DE3), and mice were immunized with the recombinant PAT protein. After cell fusion and cloning, two hybridoma cells (PATmAb-7 and PATmAb-12) were chosen since the monoclonal antibodies (Mabs) produced by them were confirmed to be specific to PAT protein in the indirect enzyme-linked immunsorbent assay (ELISA) and western blot tests. There were no cross-reactions of either Mabs to other GM proteins or to the extracts of non-GM maize. The ELISA based on the PATmAb-7 can sensitively detect 0.3 ng/g PAT protein in corn. These results indicate that the developed Mabs can be used as bio-receptors in the development of immunosensors and biosensors for the rapid and simple detection of GM corn adulterated in foods.

Seroprevalence of Dengue Virus Antibody in Korea (한국인에서 뎅기바이러스 항체의 혈청 유병률 연구)

  • Lee, Ji Hyen;Kim, Han Wool;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.132-140
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    • 2018
  • Purpose: The number of dengue fever cases is rising due to increasing overseas travel. Vaccination makes severe dengue fever in seronegative individuals after vaccination when they exposure to wild-type dengue virus. We investigated the seroepidemiology of the dengue virus for monitoring of Korean dengue virus immunity and establishing the prevention of dengue infection. Methods: The study was based on 446 residual sera collected from 98 infants (2 months to 1 year old), 152 adolescents (13 to 19 years old), 90 adults (20 to 50 years old), and 106 elderly participants (more than 65 years old) for other studies. Antibody levels for dengue virus immunoglobulin G (IgG) in each age group were measured using an enzyme-linked immunosorbent assay (ELISA). For each dengue virus IgG positive or equivocal result, an IgG ELISA was performed for Japanese encephalitis virus. Results: Of the 446 serum samples, only 1 (0.2%) adolescent had a positive result from the dengue IgG antibody test. In the dengue virus IgG antibody test, 14 (3.1%) samples showed equivocal results (10 adolescents and 4 elderly). In the 1 positive case of dengue virus IgG, the Japanese encephalitis IgG test was also positive. In the 14 equivocal cases of dengue virus IgG, there were 6 positive, 3 equivocal, and 5 negative of Japanese encephalitis IgG. Conclusions: The seroprevalence rate of dengue virus was very low in Koreans. This study provides important data for establishing the policy for preventive measures of dengue fever. It will be necessary to continuously monitor for dengue virus immunity.

RNA Interference of Chitinase Gene in Spodoptera litura (담배거세미나방(Spodoptera litura) Chitinase gene의 RNA interference)

  • Jeon, Mi Jin;Seo, Mi Ja;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.202-209
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    • 2014
  • RNA interference (RNAi) is the method which controls phenotypes of gene in live cells. Chitinase is the enzyme helping digestion and absorption of old cuticles during the ecdysis of insects. In order to investigate molting-inhibition effect with the chitinase related gene in Spodoptera litura, RNA was extracted from the $5^{th}$ instars. cDNA was synthesized and then we obtained about 700 bp size chitinase. After PCR products were cloned into a pGEM T-easy vector, colonies were picked. DNA was extracted from the colony cultures. EcoR I enzyme was used to check whether PCR products were inserted or not. And then we confirmed vector band of about 3 kb and insert band of about 700 bp. To synthesize the dsRNA, each DNA was cut with Spe I and Nco I enzymes (Circular DNA became lineared DNA). After synthesis of dsRNA, approximately 5 ul dsRNA was injected into the $3^{rd}$ abdominal segment of S. litura $4^{th}$ larvae. The concentration of dsRNA was about $10{\mu}g/{\mu}l$. We confirmed larval-larval molting : there were phenotypically abnormal individuals - for instance malformation, molting inhibition and change of integument color. Pupaadult molting : there were phenotypically abnormal individuals - for instance molting inhibition, change of wings and malformation. Also we could investigate the pupation, emergence and variation about noninjection, treated with DW and dsRNA. Each pupation was non-injection 83.3%, DW 78.3% and dsRNA 66.7%. Each emergence was non-injection 90.0%, DW 72.3% and dsRNA 65.0%. So we considered that chitinase dsRNA induced molting inhibition effect. But each variation was non-injection 8.9%, DW 2.9% and dsRNA 19.2%. Therefore dsRNA group showed the highest variation value. When 18 hours after injecting dsRNA, we could obtain abnormal individual.

Difference of Protoporphyrin IX Accumulation and Antioxidative Activity of Wheat and Barley by Protoporphyrinogen Oxidase-Inhibiting Herbicides (Protoporphyrinogen Oxidase 저해형 제초제에 대한 밀과 보리의 Protoporphyrin IX축적 및 항산화 방어계 차이)

  • 국용인;구자옥;천상욱
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.1
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    • pp.79-88
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    • 1997
  • This experiment was conducted to investigate the protoporphyrin Ⅸ (PPIX)accumulation, activity of antioxidative enzymes and contents of antioxidant in tolerant-wheat and susceptible-barley to protoporphyrinogen oxidase (Protox) inhibiting-herbicides [oxyfluorfen(2-chloro-l-(3-ethoxy-nitrophenoxy-4-(trifluoromethyl) benzene, acifluorfen (5-[2-chloro-4-(trifl-uoromethyl) phenoxy]-2-nitrobenzoic acid), bifenox(methyl-5-(2, 4-dichlorophenoxy) 2-nitroben-zoate), and oxadiazon (5-tert-butyl-3-(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one)]. The tolerant-wheat and susceptible-barley were soaked in these compounds at 10$^{-6}$ M for 2hrs and exposed to light for 2,4,6 or 8hrs to investigate change of the activity of antioxidative enzymes. The activities of monodehydroascorbate reductase(MDAR), catalase(CAL) and superoxide dismutase(SOD) were lower in the barley than in the wheat after the treatement of these compounds. The activity of peroxidase(POX) was lower in the barley than in the wheat at 8hrs after the treatment of oxyfluorfen but other compounds showed no difference in activity in wheat and barley. The activity of glutathione reductase(GR) was increased in wheat and barley according as hours of treatment of these compounds became increased but its activity was no difference between wheat and barley. In the case of the content of vitamin C due to the treatment of these compounds, the wheat decreased less than the barley. After the treatment of oxyfluorfen the content of vitamin E in the wheat was higher than in the barley but other compounds didn't have any difference between wheat and barley. And after the treatment of acifluorfen the content of carotenoid was greater in the wheat than in the barley but other compounds didn't have any difference between wheat and barley. The content of glutathione (GSH, GSSG) was greater in the barley than in the wheat. The content of protoporphyrin Ⅸ (PPIX) accumulation by the treatments of these compounds was more in the barley than in the wheat. Especially, the treatment of oxyfluorfen and acifluorfen were more accumulated 2.3 and 1.3 fold in the barley than in the wheat, respectively.

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The Clinical Study of the Lower Respiratory Tract Infection by Respiratory Syncytial Virus on Children under 2 Year of Age (2세 이하 어린이에서 Respiratory Syncytial Virus에 의한 하기도 감염에 관한 연구)

  • Cho, Jung Ik;Choi, Hyung Chul;Kim, Jong Duck;Cho, Ji Hyun
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.193-200
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    • 2000
  • Purpose : This study was designed and performed for evaluations of clinical manifestation and course of the children under 2 year of age with respiratory tract infection and positive respiratory syncytial virus(RSV) antigen. Methods : The selection criteria of the patients were children under 24 month-of-age, Clinical manifestation of respiratory tract infection, and positive RSV antigen that was detected by Vitek ImmunoDiagnostic Assay System(VIDAS) from nasal cavity. The additional laboratory and simple chest X-ray findings were reviewed from the chart of children who were admitted Wonkwang university hospital from October 1999 to March 2000. Results : Total number of patients enrolled on this study was 102. The 48(47%) children were RSV antigen positive by VIDAS method. Abnormal chest X-ray findings were noticed in 38 cases. The male to female sex ratio of 48 RSV antigen positive cases was 1.2 : 1. The mean and range of age was $10.2{\pm}5.9$ and 1.0~24 months. The peak outbreak of cases was noticed on November, 1999. All of the cases shows coughing but rale was audible in 30 cases(60%). Dyspnea, wheezing, and intercostal retraction were noticed 11(23%), 15(31%), and 10(21%) cases respectively. The most common chest X-ray finding was scattered patch infiltration that was noticed in 30 cases(63%). The mean total white blood cell counts in peripheral blood was $12,608{\pm}4,686/mm^3$. The mean blood level of IgA and IgE were $50.8{\pm}20.9$ and $72.1{\pm}98.3mg/dL$ respectively. The C-reactive protein was $16.0{\pm}18.5mg/L$. Total 5 cases need a mechanical respiraton. The duration of admission was under 7 days in 36 cases(75%). Conclusion : The RSV antigen was detected commonly in late fall and winter season. The severity of children under 2 years old with RSV respiratory tract infection take in some degree a gave courses.

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Mycelial Culture of Lentinus edodes Alleviates Rat Liver Toxicity Induced by Carbon Tetrachloride and Ethanol (표고버섯균사체의 사염화탄소 및 알콜로 처리된 흰쥐 간기능 보호 효과)

  • Ha, Yeong-L.;Kim, Young-S.;Ahn, Chae-R.;Kweon, Jung-M.;Park, Cherl-W.;Ha, Young-K.;Kim, Jeong-O.
    • Journal of Life Science
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    • v.20 no.1
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    • pp.133-141
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    • 2010
  • The protective effect of a mixed powder from solid-cultured and liquid-cultured Lentinus edodes mycelia (2:1, w/w) (designate LED) on the carbon tetrachloride ($CCl_4$)- and ethanol-induced hepatotoxicity of male Sprague-Dawley (SD) rat was investigated. In the $CCl_4$-induced rat hepatotoxicity experiment, rats of 4 groups (6 rats/group) were administere with Normal (0.2 ml distilled water), Control (0.2 ml distilled water), LED (LED 200 mg/kg BW + 0.2 ml distilled water), and Silymarin (200 mg/kg BW + 0.2 ml distilled water), p.o., daily for 2 weeks. Afterwards, all groups except for the Normal group were subjected to abdominal injection with $CCl_4$ ($CCl_4$ : corn oil, 1:1 v/v; 0.5 ml/kg BW). For the ethanol- induced rat hepatotoxicity experiment, rats were divided into 5 groups (5 rats/group): Normal; Pair-fed control (PFC); Control (ethanol); LED (ethanol + LED 200 mg/kg BW); and Silymarin (ethanol + silymarin 200 mg/kg BW). Rats of the Normal and PFC groups were fed a basal liquid diet, and rats of the Control, LED, and Silymarin groups were fed a liquid ethanol diet containing LED or Silymarin. Eight weeks later, blood and liver samples were collected to analyze biomarkers. In $CCl_4$-induced SD rats, LED elevated hepatic superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH peroxidase) activities and thiobarbituric reactive substances (TBARS) were reduced, resulting in the reduction of glutamate-oxalate transaminase (GOT), glutamate-pyruvate transaminase (GPT) and lactic dehydrogenase (LDH) activities in plasma. Similar results of these enzymes and biochemical markers in both liver tissues and plasma were seen in ethanol-induced hepatotoxicity of SD rats. In addition, elevated alcohol dehydrogenase (ADH) activity and reduced expression of cytochrome p450 mixed monooxygenase enzyme (CYP2E1) were seen in liver tissues from ethanol-treated rats by LED treatment. These effects of LED were similar to those of Silymarin. In in vitro experiments, LED showed antioxidant activity in a 2,2-diphenyl-1-picrylhydrazyl (DPPH) system and mouse liver mitochondria system induced by NADPH/$Fe^{2+}$ and cumine hydroperoxide (CuOOH). These results indicate that LED protected SD rat hepatotoxicity, induced by $CCl_4$ and ethanol, through its antioxidative activity and might be useful as a material for protection from hepatoxicity in humans.

Carbon Monoxide Inhibits PMA-induced Differentiation in Human Monocytic THP-1 Cells (단핵구세포주 THP-1의 대식세포로의 분화 및 활성화에서 CO의 억제 효과)

  • Kim, Da Sol;Lee, Mi Sun;Kim, Han Sol;Lee, Hye Yun;Kim, Oh Yun;Kang, Ye Rin;Sohn, Dong Hyun;Kim, Koanhoi;Park, Young Chul
    • Journal of Life Science
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    • v.27 no.2
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    • pp.217-224
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    • 2017
  • Carbon monoxide (CO), a reaction product of cytoprotective enzyme heme oxygenase-1 (HO-1), is a gaseous messenger with anti-proliferative, anti-apoptotic, and anti-inflammatory actions in many cell types. Here, we investigated the role of CO on the process of monocyte differentiation induced by phorbol 12-myristate 13-acetate (PMA) in human monocytic THP-1 cells. CORM-2 (tricarbonyldichlororuthenium (II) dimer, $Ru2Cl_4(CO)_6$), a CO-releasing compound, decreased a marked cell adherence with a slight reduction of proliferation in monocytic THP-1 cells treated with PMA. And, CORM-2 significantly inhibited expression of differentiation markers such as CD14, CD11b plus CD18 (macrophage-1 antigen, Mac-1 or complement receptor 3, CR3) and phagocytosis of carboxylate-modified red fluorescent latex beads, in PMA-stimulated THP-1 cells. For the further experiments, differentiation of PMA-treated cells was enhanced after the initial 2 days stimulus by removing the PMA-containing media then incubating the cells in fresh media for a another 4 days. And, we observed the secretion of inflammatory cytokines and phagocytosis in differentiated macrophages. Treatment with CORM-2 significantly abolished the secretion of IL-6, $TNF-{\alpha}$ and phagocytosis using fluorescence-conjugated E. coli (K-12 strain) bioparticles in lipopolysaccharide (LPS)-stimulated differentiated macrophages. In conclusion, these results suggest that CO inhibits the differentiation of monocytic THP-1 cells as well as the activation of differentiated macrophages.

Anti-Inflammatory Effects of Picrasma Quassioides (D.DON) BENN Leaves Extracts (소태나무 잎 추출물의 항염증 효과)

  • Jung, Yeon Seop;Eun, Cheong Su;Jung, Young Tae;Kim, Hyun Jeong;Yu, Mi Hee
    • Journal of Life Science
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    • v.23 no.5
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    • pp.629-636
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    • 2013
  • This study was performed to evaluate the anti-inflammatory and antioxidant activities of methanol extract from the leaves of Picrasma quassioides BENNET (PLME). The antioxidant effects of PLME were measured based on polyphenol and flavonoid contents. PLME was found to have $367.52{\mu}g/mg$ and $46.61{\mu}g/mg$ high polyphenol and flavonoid contents. Cell viability was determined by MTT assay. The production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) was measured by Griess assay and enzyme-linked immunosorbent assay (ELISA). In order to effectively anti-inflammatory agents, we examined the inhibitory effects on the production of lipopolysaccharide (LPS)-induced NO and $PGE_2$ in RAW 264.7 cells. PLME significantly decreased the production of NO and $PGE_2$ in a dose-dependent manner, and also reduced the expression of iNOS, a COX-2 protein. In addition, PLME reduced the NF-${\kappa}B$, $I{\kappa}B$ phosphorylation in RAW 264.7 cells upon stimulation with LPS (100 ng/ml) for 24 h. These results provide evidence for the anti-inflammatory and antioxidant effects of Picrasma quassioides leaves.

Effects of the Deer Antler Extract on Scopolamine-induced Memory Impairment and Its Related Enzyme Activities (녹용 추출물이 치매 동물모델의 기억력 개선과 관련효소 활성에 미치는 효과)

  • Lee, Mi-Ra;Sun, Bai-Shen;Gu, Li-Juan;Wang, Chun-Yan;Fang, Zhe-Ming;Wang, Zhen;Mo, Eun-Kyoung;Ly, Sun-Young;Sung, Chang-Keun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.4
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    • pp.409-414
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    • 2009
  • The aim of this study was to investigate the ameliorating effects of deer antler extract on the learning and memory impairments induced by the administration of scopolamine (2 mg/kg, i.p.) in rats. Tacrine was used as a positive control agent for evaluating the cognition enhancing activity of deer antler extract in scopolamine-induced amnesia models. The results showed that the deer antler extract-treated group (200 mg/kg, p.o.) and the tacrine-treated group (10 mg/kg, p.o.) significantly ameliorated scopolamine-induced amnesia based on the Morris water maze test. Although there was no statistical significance of brain ACh contents among the experimental groups, the brain ACh contents of the deer antler extract-treated group was slightly higher than that of the scopolamine-treated group. The inhibitory effect of deer antler extract on the acetylcholinesterase activity in the brain was significantly lower than that of scopolamine-treated group. The tacrine- and the deer antler-treated groups reduced the MAO-B activity compared to the scopolamine-treated group, but not significantly. These results suggest that the deer antler extract could be an effective agent for the prevention of the cognitive impairment induced by cholinergic dysfunction.

Effects of Probiotic Complex on Performance, Blood Biochemical and Immune Parameters, Digestive Enzyme Activity, Fecal Microbial Population and Noxious Gas Emission in Broiler Chicks (복합생균제가 육계의 생산성, 혈액생화학성분과 면역지표, 소화효소 활성도, 분중 미생물 및 유해가스 발생에 미치는 영향)

  • Kim, Min-Jeong;Jeon, Dong-Gyung;Ahn, Ho-Sung;Yoon, Il-Gyu;Moon, Eun-Seo;Lee, Chai-Hyun;Lim, Yong;Jang, In-Surk
    • Korean Journal of Poultry Science
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    • v.47 no.3
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    • pp.169-180
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    • 2020
  • This study examined the effects of a probiotic complex (PC) containing Lactobacillus plantarum, Bacillus subtilis, and Saccharomyces cerevisiae on growth performance, organ weight, immune parameters, fecal microbial count, and noxious odor in broiler chicks. A total of 216 birds (4-day-old) were fed a basal diet (CON) and a diet supplemented with 0.25% (PC1) and 0.5% (PC2) of PC until 35 days of age. No difference in body weight, feed intake, and FCR was observed among the groups. The intestinal mucosal weight of the PC1 group was greater than that of the CON group without affecting weights of the other organs. Intestinal secretory immunoglobulin A (sIgA) levels in the PC2 group increased significantly (P<0.05) compared with that in the CON group. The PC2 group also had a strong tendency for elevated blood sIgA levels. Dietary PC did not affect the level of interleukin-1β in the blood and mucosal tissues or alter maltase, sucrase, and leucine aminopeptidase activities in the intestinal mucosa. The PC2 group had higher colony-forming units (cfu) for L. plantarum and S. cerevisiae, but lower cfu for E. coli than those in the CON group. Compared to the CON diet, the PC2 diet resulted in a decreased H2S concentration and a tendency toward decreased CH3SH concentration. In conclusion, a 0.5% PC diet showed increased sIgA and desirable microbial population, and decreased noxious odor in the feces, suggesting that PC could be applied as an environmentally friendly feed additive in broiler chicks.