• Molecules and Cells
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• 제44권12호
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• pp.911-919
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• 2021

The virus-induced genome editing (VIGE) system aims to induce targeted mutations in seeds without requiring any tissue culture. Here, we show that tobacco rattle virus (TRV) harboring guide RNA (gRNA) edits germ cells in a wild tobacco, Nicotiana attenuata, that expresses Streptococcus pyogenes Cas9 (SpCas9). We first generated N. attenuata transgenic plants expressing SpCas9 under the control of 35S promoter and infected rosette leaves with TRV carrying gRNA. Gene-edited seeds were not found in the progeny of the infected N. attenuata. Next, the N. attenuata ribosomal protein S5 A (RPS5A) promoter fused to SpCas9 was employed to induce the heritable gene editing with TRV. The RPS5A promoter-driven SpCas9 successfully produced monoallelic mutations at three target genes in N. attenuata seeds with TRV-delivered guide RNA. These monoallelic mutations were found in 2%-6% seeds among M₁ progenies. This editing method provides an alternative way to increase the heritable editing efficacy of VIGE.

• 한국작물학회지
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• 제60권2호
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• pp.239-247
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• 2015

This experiment was conducted to obtain the have higher contents of pharmaceutical constituents as well as higher yield from colchicine induced diploid and tetraploid extracts of Platycodon grandiflorum. In order to determine the biological activity, this study was focused to evaluate the cytotoxicity, antimicrobial on the bronthus disease bacteria, antioxidant enzyme activity of diploid and tetraploid extracts in P. grandiflorum. The activities of antioxidant enzyme according to different solvent extracts were measured as superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX). The cytotoxicity of methanol extracts of P. grandiflorum showed significant differences between tetraploid and diploid. That is, the cytotoxic effect against human cancer cell was higher in tetraploid than in diploid. At all extracts concentration, tetraploid samples showed high toxicity and the $IC_{50}$ (concentration causing 50% cell death) value showed the highest on HCT-116 cell ($105.91{\mu}g/mL$), and exhibited significant activity against the Hep 3B cell ($140.67{\mu}g/mL$), SNU-1066 cell ($154.01{\mu}g/mL$), Hela cell ($158.37{\mu}g/mL$), SNU-601 cell ($182.67{\mu}g/mL$), Calu-6 cell ($190.42{\mu}g/mL$), MCF-7 cell ($510.19{\mu}g/mL$). Antimicrobial activities of diploid P. grandiflorum were relatively low compared to tetraploid P. grandiflorum on most of the bacterial strains. In tetraploid P. grandiflorum, K. pneumoniae showed the clear zone formation (18~19 mm) of growth inhibition, followed by the clear zone formation of 13~15 mm on C. diphtheria and S. pyogenes. The antimicrobial activities in diploid P. grandiflorum were the highest on K. pneumonia (14~15 mm), and showed the clear zone formation of 11~12 mm on C. diphtheria and 12~13 mm on S. pyogenes. The antimicrobial activity is thought to look different depending on the bacterial strains and the polyploidy of P. grandiflorum. The root extract of P. grandiflorum had the highest (97.2%) SOD enzyme activity in ethyl acetate partition layer of tetraploid while water partition layer of diploid showed the lowest (48.6%) SOD enzyme activity. The activity of CAT showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all partition layers except butyl alcohol. The activities of APX and POD showed higher values in the root of tetraploid than in the diploid of P. grandiflorum in all fraction solvents except water layer. These results indicate that the tetraploid P. grandiflorum can be used as a source for developing cytotoxic agent and antimicrobials which can act against bronchus diseases bacterial strains.

• 대한수의학회지
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• 제39권3호
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• pp.566-574
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• 1999

This study was conducted to examine the outbreak rate and the causative agents of otitis externa in 26 dogs (49 ears ; 23 dogs = bilateral, 3 dogs = unilateral), and the normal microfloras of external ear canal in 68 dogs(133 ears ; 65 dogs = bilateral, 3 dogs = unilateral ) in Taegu, 1997. The breed, living environment, sex, age and season distribution of otitic dogs were as follows : Dogs with erect and hairy ears(42.3%), pendulous and hairy ears(38.5%), indoor(92.3%), female(65.4%) and below one year old(38.5%) were more prevalent. According to season, otitis externa was mainly occurred between July and October. The major causative agents of canine otitis externa were Malassezia pachydermatis (32.7%), Staphylococcus aureus (26.5%) and S intermedius (16.3%). In the microorganism isolated 39 otitic ear canals, single infection was 53.8% and mixed infection was 46.2%. The normal microfloras of canine external ear canal were fungi including M pachydermatis, Aspergillus spp, Microsporum canis, Alternaria spp, Verticillium spp and Yeast, and bacteria including Staphylococcus spp(10 species including S xylosus), Bacillus spp, Corynebacterium spp, Listeria spp, Actinomyces pyogenes and Escherichia coli. No growth was 34.6%.

• 한국식품영양과학회지
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• 제45권7호
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• pp.1017-1025
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• 2016

기관지질환에 우수한 기능성을 가지고 있는 도라지는 수십종의 다양한 사포닌 종류를 함유하고 있다. 이러한 도라지를 젖산발효 시킬 경우 사포닌 조성의 변화와 기관지질환 유발 세균에 대한 항균 활성의 변화를 측정하였다. 도라지 발효를 위하여 사용된 젖산균은 Leuconostoc mesenteroides N12-4, Leuconostoc mesenteroides N58-5, Lactobacillus plantarum N76-10, Lactobacillus plantarum N56-12, Lactobacillus brevis N70-9 및 Lactobacillus brevis E3-8 등 6종이었다. 도라지 발효 시 젖산균 생육은 L. plantarum균이 가장 활발하였으며 pH 값도 가장 많이 내려가 발효 24시간째 pH는 3.73을 보였고 colony 수는 $10^7CFU/g$ 이상으로 증가하였다. 젖산발효 시간 동안 도라지 사포닌 9종에 대한 조성 변화는 사용된 도라지 시료에서 함량이 높았던 platycoside E, diapioplatycoside E, platycodin A, polygalacin D 등이 발효기간 중 일정량 감소하다가 발효 72시간 이후부터는 증가하는 경향이었고 platycodin D는 약간 증가하다가 72시간 이후부터는 오히려 감소하는 경향이었으며, 총사포닌 함량은 발효기간 동안 함량의 변화가 없었던 L. brevis E3-8균을 제외하고는 발효 72시간 이후 96시간째 증가하였다. 기관지질환 유발세균에 대한 항균 활성의 변화를 보기 위하여 사용된 세균은 Corynebacterium diphtheriae KCTC 3075, Klebsiella pneumoniae KCTC 2246, Staphylococcus aureus KCTC 1621 및 Streptococcus pyogenes ATCC 19615균 등 4종으로 각 균에 대한 항균 활성이 발효에 의하여 증대되는 젖산균은 L. plantarum균으로, 특히 L. plantarum N56-12균이 강한 항균 활성을 보여주었다. 이에 따라 각 기관지질환 유발세균에 대한 MIC를 조사한 결과 각 균에 대하여 도라지 무발효물은 200, 180, 210 및 90 mg/mL였고, L. plantarum N56-12균으로 발효된 도라지 상등액은 45, 10, 50 그리고 25 mg/mL였다.

• Mycobiology
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• 제40권1호
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• pp.8-13
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• 2012

In the present study, an attempt to evaluate the antimicrobial and antioxidant activity of fungal endophytes inhabiting $Emblica$ $officinalis$ has been made keeping in view the medicinal importance of the selected host plant in Indian traditional practices. A total of four endophytic fungi belonging to Phylum Ascomycetes were isolated from different parts of the plant which were characterized morphologically and by using rDNA-internal transcribed spacer. The most frequently isolated endophyte was $Phomopsis$ sp. The antioxidant activity by 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and reducing power assay, and total phenol were evaluated using ethanolic extract of endophytic fungi. DPPH activities in all the ethanolic extract increased with the increase in concentrations. Endophytes, $Phomopsis$ sp. and $Xylaria$ sp. showed highest antioxidant activity and also had the higher levels of phenolics. Antimicrobial activity of fungal extract were tested against four bacteria namely, $Escherichia$ $coli$ MTCC730, $Enteroccocus$ $faecalis$ MTCC2729, $Salmonella$ $enterica$ ser. $paratyphi$ MTCC735 and $Streptococcus$ $pyogenes$ MTCC1925, and the fungus $Candida$ $albicans$ MTCC183. In general, the fungal extracts inhibited the growth of test organisms except $E.$ $coli$.

• 한국식물병리학회지
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• 제11권2호
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• pp.158-164
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• 1995

토마토의 엽권에서 분리한 Fusariym graminearum이 분비하는 물질은 벼 도열병균(Pyricularia oryzae)의 여러 종의 식물병원 진균에 대한 항균활성을 나타내었으며, 이러한 활성물질을 PDA에서 본 균을 배양 한 후 chloroform으로 추출하여 분리정제 하였다. HPLC에 의하여 5종류의 활성 물질을 분획하였으며, 그중 1번(F402) 화합물을 벼 도열병균(P. oryzae)을 포함한 22개 식물 병원 진균에 대하여 살균 활성범위를 조사한 결과, 이 화합물은 50$\mu\textrm{g}$/ml 농도에서 Pythium ultimum, Rhizoctonia solani, Sclerotinia sclerotiorum은 전혀 억제하지 못하였으며, Phytophthora spp., Cladosporium fulvum, Fusarium spp., Corynespora cassicola에는 어느 정도의 활성이 있었지만 낮게 나타났고, P. oryzae, Cochliobolus miyabeanus, Alternaria solani는 100% 억제하여 활성이 높게 나타났다. 또한 장내 세균에 대한 활성을 MIC로 비교할 때 Streptococcus pyogenes, Streptococcus faecium에 대하여는 각각 12.5, 25 $\mu\textrm{g}$/ml였고 Staphylococcus aureus는 25-50$\mu\textrm{g}$/ml으로 나타났으며, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella aerogenes, Enterobacter cloacae에서는 100$\mu\textrm{g}$/ml 이상으로 활성이 나타나지 않았다. F402를 200$\mu\textrm{g}$/ml의 농도로 직접 살포한 식물체에서의 방제효과는 벼도열병, 벼 깨씨무늬병, 보리 흰 가루병에 대하여는 80%이상이었으나, 벼 잎집무늬마름병, 오이 잿빛곰팡이병, 토마토 역병, 밀 녹병에서는 낮았다.

• 한국동물위생학회지
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• 제19권2호
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• pp.115-125
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• 1996

A total of 315 microorgainisms were isolated from 256 pneumonic lung samples of pig in abattoirs of northern Chungnam area during February to November 1994, and Identified 97 strains as Pusteurella multocida, 89 strains as Staphylococcus spp, 54 strains as Streptococcus spp, 22 strains as Mycoplasma spp, 21 strains as Escherichia coli, 18 strains as Haemophilus parsuis, 11 strains as Corymebacteroi, pyogenes, and 3 strains as Acrinobacillus spp by Gram's and Dienes stain, and biological properties test Involved API system. After that, they were examined anti biotic susceptibility for ampicillin(AM), cephalothin(CF), chloramphenicol(CP), erythromycin(EM), kanamycin(KM), gentamicin(GM), neomycin(NM), penicillin(PC), streptomycin(SM), tetracycline(TE), tiamulin(TIA), tylosin(TYL), methicillin(DP), colistin(CL) and trimet hoprim(SXT). In antibiotics susceptibility test, 293 isolates except Mycoplasma spp 22 strain were highly susceptible to DF(79.2%) and AM(76.2%), but resistant to PC(14.0%), NM(19.5%) and KM (23.2%) The multiple drug resistant patterns were noted in most isolates, whereas only 7 isolates resistant to single drugs.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1997년도 춘계학술대회
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• pp.40-47
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• 1997

New quinolones generally have a broad antibacterial spectrum against gram-positive, gram-negative, glucose-nonfermenting and anaerobic bacteria. Some of newly developed quinolones have potent activities against S. aureus including MRSA, S.pneumoniae including PRSP, B. fragilis, chlamydiae, mycoplasmas and mycobacteria as well, and show good activities against various strains resistant to antibacterial agents of other classes. Quinolones display postantibiotic effects in vitro and are bactericidal at concentrations similar to or twice that of the minimum inhibitory concentrations (MICs) for susceptible pathogens. In experimental murine infection models including systemic infections with various pathogens such as S. aureus, S. pyogenes, S. pneumoniae, E. coli and P. aeruginosa, quinolones have shown good oral efficacy as well as parenteral efficacy. Good oral absorption and good tissue penetration of quinolones account for good therapeutic effects in clinical settings. The target of quinolones are two structurally related type II topoisomerases, DNA gyrase and DNA topoisomerase IV. Quinolones are shown to stabilize the ternary quinolone-gyrase-DNA complex and inhibit the religation of the cleaved double-stranded DNA. Bacteria can acquire resistance to quinolones by mutations of these target enzymes. Mutation sites and amino acid changes in DNA gyrase and DNA topoisomerase IV are similar in the organisms examined, suggesting that the mechanism of quinolone resistance in the target enzymes is essentially the same among various organisms. Quinolones act on both the target enzymes to different degrees depending on the organisms or agents tested, and bacteria become highly resistant to quinolones in a step-wise fashion. Incomplete cross-resistance among quinolones in some strains of E. coli and S. aureus suggests the possibility of finding quinolones active against quinolone-resistant strains which are prevailing now. To find such quinolones, the potency toward two target enzymes and the membrane permeability including influx and/or efflux systems should be taken into account.

• Clinical and Experimental Pediatrics
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• 제55권5호
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• pp.177-180
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• 2012

OK-432 (picibanil) is an inactivated preparation of $Streptococcus$ pyogenes that causes pleurodesis by inducing a strong inflammatory response. Intrapleural instillation of OK-432 has recently been used to successfully treat neonatal and fetal chylothorax. Here we report a trial of intrapleural instillation of OK-432 in two preterm infants who were born with hydrops fetalis and massive bilateral pleural effusion. Both cases showed persistent pleural effusion, refractory to conservative treatment, up to postnatal days 26 and 46, respectively. An average of 80 to 140 mL of pleural fluid was drained daily. In case 1, the infant was treated with OK-432 during the fetal period at gestation 28 weeks and 4 days of gestation, but showed recurrence of pleural effusion and progressed into hydrops. Within two to three days after OK-432 injection, the amount of pleural fluid drainage was dramatically decreased and there was no reaccumulation. We did not observe any side effects related to OK-432 injection. We suggest that OK-432 should be considered as a therapeutic option in infants who have persistent pleural effusion for more than four weeks, with the expectation of the early removal of the chest tube and a good outcome.

• Journal of Periodontal and Implant Science
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• 제38권1호
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• pp.31-40
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• 2008

Purpose: The probiotic effects of lactic acid bacteria have widely been researched in diverse human pathogens, but only a few effects are reported against oral pathogens. The antimicrobial effects of the Enterococcus faecium 7413 isolated from Korean infants on the 9 pathogen including 6 oral streptococci were investigated the clinical use of the antimicrobial peptide for oral microflora control. Materials and Methods: E. faecium 7413 was identified by morphological, biochemical tests and 16S rDNA sequence analysis. Inhibitory effects of culture supernatants were determined for their ability to grow on agar plate containing pathogenic bacteria. Result: The culture supernatant of Enterococcus faecium 7413 showed inhibitory effects on oral pathogens, namely Streptococcus pyogenes KCTC 3556, S. pneumoniae KCTC 5080, S. mutans ATCC 25175, S. anginosus ATCC 33397, S. constellatus KCTC 3268, S. intermedius ATCC 27823 and Shigella flexneri KCTC 2008. Whereas it did not affect the multiplication of E. coli strains, KCTC 1041 and ATCC 43894. Conclusion: The data obtained in this study could be useful for future development of effective probiotics allowing prevention for oral pathogens.