• Title/Summary/Keyword: bio assay

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Studies on Phytotoxin in Intensively Cultivated Upland Crops -I. Identification of phytotoxin in soil and effects of phytotoxin application to the toxicity of hot-pepper plant (연작재배지토양(連作栽培地土壤)의 식물독소(植物毒素)에 관(關)한 연구(硏究) -제(第)1보(報) 토양중(土壤中) 식물독소(植物毒素)의 분리정량(分離定量) 및 식물독소(植物毒素) 첨가(添加)가 고추 유식물(幼植物)에 미치는 영향(影響))

  • Lee, Sang Kyu;Suh, Jang Sun;Kim, Young Sig;Park, Jun Kyu
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.1
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    • pp.63-67
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    • 1987
  • A laboratory experiment was conducted to find out the concentration of phytotoxin in intensively cultivated hot-pepper, garlic and chinese cabbage, and effects of these phytotoxin to the germination and growth of young hot-pepper plant. Also this experiment presents describes of the bio-assay method and results of phytotoxin application to the toxicity of hot-pepper plant. The results obtained were summarized as follows; 1. A series of non-volatile (aromatic) phenolic compounds such as hydroquinone, benzoic-, p-hydroxybenzoic, and vanillic acid were quantitatively and qualitatively analysed using BSA(N, O-bis(trimethylsilyl)acetamide) by means of gas chromatography method. 2. Phytotoxin as hydroquinone, benzoic-, p-hydroxybenzoic- and vanillic acid were determined in intensively cultivated hot- pepper, garlic and chinese cabbage. Highest concentration of phytotoxin was obtained in hot-pepper cultivated soil. 3. Direct toxic action of the applied phytotoxin to the germination and young hot-pepper plant growth was observed at the levels of 200 ppm. Benzoic acid was obtained the highest toxicity to the young hot-pepper plant growth. 4. Mode of actions of phytotoxins to the young hot-pepper plant growth were observed as stunting of stem elongation, discoloration of leaf and oxygen depletion from consideration as causes of symptom.

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Application of Oral Fluid Sample to Monitor Porcine circovirus-2 Infection in Pig Farms (구강액을 이용한 양돈장의 Porcine circovirus-2 감염에 대한 모니터링)

  • Kim, Won-Il
    • Journal of Veterinary Clinics
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    • v.27 no.6
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    • pp.704-712
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    • 2010
  • Porcine circovirus-2 (PCV2) has been implicated in many clinical diseases/syndromes that are now referred to as PCV-associated diseases (PCVAD). Due to significant economic losses caused by PCVAD, many swine operations have launched extensive monitoring programs for PCV2. Traditional serum sampling is, however, rather expensive and laborious, hampering effective large scale pathogen surveillance. A field-based longitudinal study was conducted to assess the utility of pen-based oral fluid sample as an alternative to serum for herd PCV2 testing. Six pens (25 pigs/pen) at each of 3 different sites were used in the study. One oral fluid and 5 random serum samples per pen were collected at 3, 5, 8, 12, and 16 weeks of age, and the sera were pooled by pen for testing. All samples were tested for PCV2 by real-time PCR and for antibodies by indirect fluorescent antibody test (for both anti-PCV2 IgG and IgA) and 3 ELISA assays (blocking ELISA, indirect ELISA, and IgG/IgM sandwich ELISA). PCV2 DNA was detected in oral fluid samples sporadically until 8 weeks and in all pens at 16 weeks. PCV2-specific IgG was detected in oral fluid samples at 3 weeks and persisted until 5 to 8 weeks in all sites. Anti-PCV2 IgG and IgA were detectable in oral fluid samples collected at 16 weeks from all of the pens at 1 site. The detection of PCV2 and anti-PCV2 antibody in oral fluid samples correlated positively with results on pooled sera, suggesting that oral fluids can be a cost-effective alternative to serum for herd monitoring of PCV2 infection.

Antioxidant Activities of Ethanol and Water Extracts from Propolis (프로폴리스 에탄올 및 물 추출물의 항산화 활성)

  • Jeong, Chang-Ho;Shin, Chang-Sik;Bae, Young-Il;Shim, Ki-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.12
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    • pp.1725-1730
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    • 2010
  • To obtain basic information on the potential use of propolis as a raw material in functional food, proximate composition, total phenolics content and antioxidant activities of different propolis extracts in Korea were investigated. Propolis had the highest level of crude fat and the lowest level of crude fiber. The total phenolics content of ethanol and water extract of propolis from Geochang (GEE and GWE), ethanol and water extract of propolis from Jeju (JEE and JWE) were 184.17, 316.19, 204.33 and 47.83 mg gallic acid equivalent/g, respectively. GWE contained relatively higher levels of total phenolics than the other extracts. The antioxidant potential of the extracts was assessed by different in vitro assays such as DPPH, ABTS, reducing power, ferric reducing/antioxidant power (FRAP) and peroxidation inhibiting activities through linoleic acid emulsion system. DPPH and ABTS radical scavenging activities of all the extracts were dose dependent. The GWE exhibited the best performance in reducing power, FRAP, and lipid peroxidation using ferric thiocyanate (FTC) assay. These results demonstrated that GWE has excellent antioxidant activities and thus it has great potential as a raw material for functional food.

Bioconverted Jeju Hallabong tangor (Citrus kiyomi × ponkan) peel extracts by cytolase enhance antioxidant and anti-inflammatory capacity in RAW 264.7 cells

  • Chang, Yun-Hee;Seo, Jieun;Song, Eunju;Choi, Hyuk-Joon;Shim, Eugene;Lee, Okhee;Hwang, Jinah
    • Nutrition Research and Practice
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    • v.10 no.2
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    • pp.131-138
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    • 2016
  • BACKGROUND/OBJECTIVES: Citrus and its peels have been used in Asian folk medicine due to abundant flavonoids and usage of citrus peels, which are byproducts from juice and/or jam processing, may be a good strategy. Therefore, the aim of this study was to examine antioxidant and anti-inflammatory effects of bioconversion of Jeju Hallabong tangor (Citrus kiyomi ${\times}$ ponkan; CKP) peels with cytolase (CKP-C) in RAW 264.7 cells. MATERIALS/METHODS: Glycosides of CKP were converted into aglycosides with cytolase treatment. RAW 264.7 cells were pre-treated with 0, 100, or $200{\mu}g/ml$ of citrus peel extracts for 4 h, followed by stimulation with $1{\mu}g/ml$ lipopolysaccharide (LPS) for 8 h. Cell viability, DPPH radical scavenging activity, nitric oxide (NO), and prostagladin $E_2$ ($PGE_2$) production were examined. Real time-PCR and western immunoblotting assay were performed for detection of mRNA and/or protein expression of pro-inflammatory mediators and cytokines, respectively. RESULTS: HPLC analysis showed that treatment of CKP with cytolase resulted in decreased flavanone rutinoside forms (narirutin and hesperidin) and increased flavanone aglycoside forms (naringenin and hesperetin). DPPH scavenging activities were observed in a dose-dependent manner for all of the citrus peel extracts and CKP-C was more potent than intact CKP. All of the citrus peel extracts decreased NO production by inducible nitric oxide synthase (iNOS) activity and $PGE_2$ production by COX-2. Higher dose of CKP and all CKP-C groups significantly decreased mRNA and protein expression of LPS-stimulated iNOS. Only $200{\mu}g/ml$ of CKP-C markedly decreased mRNA and protein expression of cyclooxygenase-2 in LPS-stimulated RAW 264.7 cells. Both 100 and $200{\mu}g/ml$ of CKP-C notably inhibited mRNA levels of $interleukin-1{\beta}$ ($IL-1{\beta}$) and IL-6, whereas $200{\mu}g/ml$ CKP-C significantly inhibited mRNA levels of $TNF-{\alpha}$. CONCLUSIONS: This result suggests that bioconversion of citrus peels with cytolase may enrich aglycoside flavanones of citrus peels and provide more potent functional food materials for prevention of chronic diseases attributable to oxidation and inflammation by increasing radical scavenging activity and suppressing pro-inflammatory mediators and cytokines.

Clinical usefulness of rapid antigen test to detect respiratory syncytial virus infection (Respiratory syncytial virus 감염진단을 위한 신속항원검사의 유용성)

  • Kim, Hyung Su;Kim, Hee La;Park, Ki Hyung;Cho, Kyung Soon
    • Clinical and Experimental Pediatrics
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    • v.51 no.10
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    • pp.1071-1076
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    • 2008
  • Purpose : Respiratory syncytial virus (RSV) is the most frequent cause of lower respiratory infections in infants and young children. Early detection allows quarantining of infected inpatients to prevent nosocomial transmission and to choose a treatment. To achieve rapid reporting, to facilitate prompt antiviral therapy, and to avoid unnecessary use of antibiotics, an easy, rapid diagnostic method for RSV is needed. We evaluated a lateral flow immunochromatography (RSV Respi-Strip test) and EIA (Enzyme immuno assay) compared to RT-PCR. Methods : From April 2007 to March 2008, 112 consecutive respiratory specimens (nasopharyngeal aspirates, throat swabs, tracheal aspirates, sputum) from patients who were suffering from the clinical signs and symptoms of respiratory tract infection were enrolled in Busan. A total of 112 patients were tested with RSV Respi-Strip (Corio-BioConcept, Belgium), EIA, and RT-PCR at the same time. Results : Of the 112 specimens tested, the number of children who showed positive results at RT-PCR and Respi-Strip were 45 and 42, respectively. The Respi-Strip rapid antigen test had a sensitivity of 88% and a specificity of 94%. The positive and negative predictive values were 90% and 92%, respectively. The agreement was 83%. Conclusion : In our study, the rapid antigen test had as much sensitivity as any method for detection of RSV. The test has many advantages such as easy performance, simple interpretation, and rapid results. If the rapid antigen test is widely applied in the clinical setting, the may be useful for diagnostic and epidemiological studies of RSV infection.

Development of herbicide-tolerant Korean rapeseed (Brassica napus L.) cultivars (한국 고유의 품종을 이용한 제초제 저항성 유채 개발)

  • Kim, Hyo-Jin;Lee, Hye-Jin;Go, Young-Sam;Roh, Kyung-Hee;Lee, Young-Hwa;Jang, Young-Seok;Suh, Mi-Chung
    • Journal of Plant Biotechnology
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    • v.37 no.3
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    • pp.319-326
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    • 2010
  • An interest in the production of seed-oil based fuel and raw materials, which comes from renewable plant sources, has been intrigued by the phenomenon of global warming and shortage of fossil fuels. Rapeseed (Brassica napus) is the most important oilseed crop, which produces seeds with 40% oil. It is desirable to develop genetically modified rapeseed producing oils, which can be easily converted to biodiesel. As an initial step for development of genetically modified rapeseed for the production of biofuels or bio-based materials, Korean rapeseed cultivars, Naehan, Youngsan, Tammi and Halla, were analyzed. Four Korean rapeseed cultivars produce 32 to 40% oil of seed dry weight, which is rich in oleic acid (more than 60 mole%). The cotyledonary petioles of rapeseed cultivar, Halla, were transformed using Agrobacterium tumefaciens strain GV3101, carrying the uidA gene encoding $\beta$-glucuronidase (GUS) as a reporter gene and the phosphinothricin acetyltransferase (PAT) gene as a selectable marker. The stable integration of PAT gene in the genome of transgenic rapeseeds was confirmed by PCR analysis. Expression of uidA gene in various rapeseed organs was determined by fluorometric assay and histochemical staining. Transformation efficiency of a Korean rapeseed Halla cultivar was 10.4%. Genetic inheritance of transgenes was confirmed in $T_2$ generation.

Methoxychlor Produces Many Adverse Effects on Male Reproductive System, Kidney and Liver by Binding to Oestrogen Receptors

  • Kim, Dae Young
    • Journal of Embryo Transfer
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    • v.28 no.2
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    • pp.157-162
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    • 2013
  • Methoxychlor (MXC) was developed to be a replacement for the banned pesticide DDT. HPTE [2,2-bis (p-hydroxyphenyl)-1,1,1-trichloroethane], which is an in vivo metabolite of MXC, has strong oestrogenic and anti-androgenic effects. MXC and HPTE are thought to produce potentially adverse effects by acting through oestrogen and androgen receptors. Of the two, HPTE binds to sex-steroid receptors with greater affinity, and it inhibits testosterone biosynthesis in Leydig cells by inhibiting cholesterol side-chain cleavage enzyme activity and cholesterol utilisation. In a previous study, MXC was shown to induce Leydig cell apoptosis by decreasing testosterone concentrations. I focused on the effects of MXC on male mice that resulted from interactions with sex-steroid hormone receptors. Sex-steroid hormones affect other organs including the kidney and liver. Accordingly, I hypothesised that MXC can act through sex-steroid receptors to produce adverse effects on the testis, kidney and liver, and I designed our experiments to confirm the different effects of MXC exposure on the male reproductive system, kidney and liver. In these experiments, I used pre-pubescent ICR mice; the puberty period in ICR mice is from postnatal day (PND) 45 to PND60. I treated the experimental group with 0, 100, 200, 400 mg MXC/kg b.w. delivered by an intra-peritoneal injection with sesame oil used as vehicle for 4 weeks. At the end of the experiment, the mice were sacrificed under anaesthesia. The testes and accessory reproductive organs were collected, weighed and prepared for histological investigation. I performed a chemiluminescence immune assay to observe the serum levels of testosterone, LH and FSH. Blood biochemical determination was also performed to check for other effects. There were no significant differences in our histological observations or relative organ weights. Serum testosterone levels were decreased in a dose-dependent manner; a greater dose resulted in the production of less testosterone. Compared to the control group, testosterone concentrations differed in the 200 and 400 mg/kg dosage groups. In conclusion, I observed markedly negative effects of MXC exposure on testosterone concentrations in pre-pubescent male mice. From our biochemical determinations, I observed some changes that indicate renal and hepatic failure. Together, these data suggest that MXC produces adverse effects on the reproductive system, kidney and liver.

Development of an In Vitro Test System Measuring Transcriptional Downregulatory Activities on IL-13

  • Choi, Jeong-June;Park, Bo-Kyung;Park, Sun-Young;Yun, Chi-Young;Kim, Dong-Hee;Kim, Jin-Sook;Hwang, Eun-Sook;Jin, Mi-Rim
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.331-337
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    • 2009
  • Interleukin-13 (IL-13) has been proposed as a therapeutic target for bronchial asthma as it plays crucial roles in the pathogenesis of the disease. We developed an in vitro test system measuring transcriptional downregulatory activities on IL-13 as a primary screening method to select drug candidates from natural products. The promoter region of IL-13 (-2,048 to +1) was cloned into the upstream of a luciferase gene in the plasmid pGL4.14 containing the hygromycin resistance gene as a selection marker, generating pGL4.14-IL-13. The EL-4 thymoma and RBL-2H3 mast cells transiently expressing this plasmid highly produced the luciferase activities by responding to PI (PMA and ionomycin) stimulation up to 8-fold and 13-fold compared with the control, respectively, whereas cyclosporin A, a well-known antiasthmatic agent, significantly downregulated the activities. The BF1 clone of RBL-2H3 cells constitutively expressing pGL4.14-IL-13 was established by selecting surviving cells under a constant lethal dose of hygromycin treatment. The feasibility of this system was evaluated by measuring the downregulatory activities of 354 natural products on the IL-13 promoter using the BF1 clone. An extract from Morus bombycis (named TBRC 156) significantly inhibited PI-induced luciferase activities and IL-13 mRNA expression, but not the protein expression. Fisetin (named TBRC 353) inhibited not only PI-induced luciferase activities and mRNA expression, but also the IL-13 protein secretion, whereas myricetin (named TBRC 354) could not suppress the IL-13 expression at all. Our data indicated that this in vitro test system is able to discriminate the effects on IL-13 expression, and furthermore, that it might be suitable as a simple and time-saving primary screening system to select antiasthmatic agents by measuring transcriptional activities of the IL-13 promoter.

A Bacterial Metabolite, Compound K, Induces Programmed Necrosis in MCF-7 Cells via GSK3β

  • Kwak, Chae Won;Son, Young Min;Gu, Min Jeong;Kim, Girak;Lee, In Kyu;Kye, Yoon Chul;Kim, Han Wool;Song, Ki-Duk;Chu, Hyuk;Park, Byung-Chul;Lee, Hak-Kyo;Yang, Deok-Chun;Sprent, Jonathan;Yun, Cheol-Heui
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.1170-1176
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    • 2015
  • Ginsenosides, the major active component of ginseng, are traditionally used to treat various diseases, including cancer, inflammation, and obesity. Among these, compound K (CK), an intestinal bacterial metabolite of the ginsenosides Rb1, Rb2, and Rc from Bacteroides JY-6, is reported to inhibit cancer cell growth by inducing cell-cycle arrest or cell death, including apoptosis and necrosis. However, the precise effect of CK on breast cancer cells remains unclear. MCF-7 cells were treated with CK ($0-70{\mu}M$) for 24 or 48 h. Cell proliferation and death were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry assays, respectively. Changes in downstream signaling molecules involved in cell death, including glycogen synthase kinase $3\beta$ ($GSK3\beta$), $GSK3\beta$, $\beta$-catenin, and cyclin D1, were analyzed by western blot assay. To block $GSK3\beta$ signaling, MCF-7 cells were pretreated with $GSK3\beta$ inhibitors 1 h prior to CK treatment. Cell death and the expression of $\beta$-catenin and cyclin D1 were then examined. CK dose- and time-dependently inhibited MCF-7 cell proliferation. Interestingly, CK induced programmed necrosis, but not apoptosis, via the $GSK3\beta$ signaling pathway in MCF-7 cells. CK inhibited $GSK3\beta$ phosphorylation, thereby suppressing the expression of $\beta$-catenin and cyclin D1. Our results suggest that CK induces programmed necrosis in MCF-7 breast cancer cells via the $GSK3\beta$ signaling pathway.

Nanopharmaceutical Approach for Enhanced Anti-cancer Activity of Betulinic Acid in Lung-cancer Treatment via Activation of PARP: Interaction with DNA as a Target -Anti-cancer Potential of Nano-betulinic Acid in Lung Cancer-

  • Das, Jayeeta;Samadder, Asmita;Das, Sreemanti;Paul, Avijit;Khuda-Bukhsh, Anisur Rahman
    • Journal of Pharmacopuncture
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    • v.19 no.1
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    • pp.37-44
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    • 2016
  • Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.