• Journal of Pharmaceutical Investigation
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• v.30 no.3
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• pp.151-158
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• 2000

Gel and cream containing 5% and 10% ketonic fraction (KF) of Leptospermum scoparium, respectively were formulated. Antimicrobial activity, stability, anti-inflammatory effect, rheological properties, drug release and acute toxicity for these topical efficacy were evaluated. Gel and cream containing neomycin or gentamycin in combination with KF has potent antimicrobial activity. Gel and cream were physically stable and did not show any creaming for 6 months storage. Gel showed plastic flow with yield value and cream showed pseudoplastic flow with hysteresis loop. The gel and cream containing KF showed higher viscosity than control or commercial one. The viscosity increased as the concentration of KF increased. Both 10% gel and cream showed a significant decrease in swelling when applied to the carrageenan- injected paw, suggesting local antiinflammatory activity. Particularly, 10% gel preparation showed similar antiinflammatory activity when compared with commercially available drugs. Percent of drug released and diffusion coefficient were in the order of 5% gel, 10% gel, 5% cream, and 10% cream, respectively. There were no significant changes of body weight in rats percutaneously administered with 10% cream and gel when compared with control. There were no induced acute toxicity when 10% cream or gel was applied to rats. Leptospermum scoparium could be practicaly used in topical preparations.

• Journal of Microbiology and Biotechnology
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• v.27 no.2
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• pp.357-364
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• 2017

In this study, we aimed to generate a series of versatile tagging plasmids that can be used in diverse molecular biological studies of the fungal pathogen Cryptococcus neoformans. We constructed 12 plasmids that can be used to tag a protein of interest with a GFP, mCherry, $4{\times}FLAG$, or $6{\times}HA$, along with nourseothricin-, neomycin-, or hygromycin-resistant selection markers. Using this tagging plasmid set, we explored the adenylyl cyclase complex (ACC), consisting of adenylyl cyclase (Cac1) and its associated protein Aca1, in the cAMP-signaling pathway, which is critical for the pathogenicity of C. neoformans. We found that Cac1-mCherry and Aca1-GFP were mainly colocalized as punctate forms in the cell membrane and non-nuclear cellular organelles. We also demonstrated that Cac1 and Aca1 interacted in vivo by co-immunoprecipitation, using $Cac1-6{\times}HA$ and $Aca1-4{\times}FLAG$ tagging strains. Bimolecular fluorescence complementation further confirmed the in vivo interaction of Cac1 and Aca1 in live cells. Finally, protein pull-down experiments using $aca1{\Delta}$::ACA1-GFP and $aca1{\Delta}$::ACA1-GFP $cac1{\Delta}$ strains and comparative mass spectrometry analysis identified Cac1 and a number of other novel ACC-interacting proteins. Thus, this versatile tagging plasmid system will facilitate diverse mechanistic studies in C. neoformans and further our understanding of its biology.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.353-359
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• 2011

Fowl cholera is a contagious acute and chronic disease caused by Pasteurella multocida in both domesticated and wild birds. Acute fowl cholera in both chickens and wild birds has recently been documented in Korea, but the chronic form has not been reported in Korea until now. This study describes the first outbreak of chronic fowl cholera in 13-week-old Arbor Acre broiler breeder chickens submitted to the College of Veterinary Medicine, Kyungpook National University in April 2006. The clinical signs of the affected flock of 9,621 chickens were lameness caused by swollen hock joints, diarrhea, ruffled feathers, and an average weekly mortality of 1.0%. At necropsy, purulent or caseous exudates were found in the hock and wing joints, humerus, and eyes, and severe pneumonia and pericarditis were discovered. Eleven bacterial isolates obtained from the liver, joint, infraorbital sinus and sternal bursa of the submitted chickens were all identified as Pasteurella multocida based on their physiological and biochemical characteristics. Five isolates were examined for antimicrobial susceptibility against 21 different antimicrobial agents including ampicillin. All were resistant to kanamycin, neomycin, and streptomycin, and some were resistant to gentamicin. The tested isolates were all susceptible to the other 17 antimicrobial agents. All 11 isolates were capsular serogroup A based on multiplex polymerase chain reaction. In addition, two of five isolates used in the antimicrobial susceptibility test were identified as somatic serotype 1 by an agar gel diffusion precipitin test, while the others were non-typable.

• BMB Reports
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• v.33 no.6
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• pp.525-530
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• 2000

The nerve growth factor (NGF) induces neuronal differentiation and neurite outgrowth of PC12 cells, whereas epidermal growth factors (EGF) stimulate growth and proliferation of the cells. In spite of this difference, NGF-or EGF-treated PC12 cells share various properties in cellular-signaling pathways. These include the activation of the phosphoinositide (PI)-3 kinase, 70 kDa S6 kinase, and in the mitogen-activated protein (MAP) kinase pathway, following the binding of these growth factors to intrinsic receptor tyrosine kinases (RTKs). Therefore, many studies have been attempted to access the critical signaling events in determining the differentiation and proliferation of PC12 cells. In this study, we investigated the cytosolic phospholipase $A_2$ ($cPLA_2$) in neurite behavior in order to identify the differences of signaling pathways between the NGF-induced differentiation and the EGF-induced proliferation of PC12 cells. We have showed here that the $cPLA_2$ was translocated from cytosol to membrane only in NGF-treated cells. We also demonstrated that this translocation is associated with NGF-induced activation of phospholipase $C-{\gamma}(PLC-{\gamma})$, which elevates intracellular $Ca^{2+}$ concentration. These results reveal that the translocation of $cPLA_2$ may be a requisite event in the neuronal differentiation of PC12 cells. Various phospholipase inhibitors were used to confirm the importance of these enzymes in the differentiation of PC12 cells. Neomycin B, a PLC inhibitor, dramatically inhibited the neurite outgrowth, and two distinct $PLA_2$ inhibitors, 4-bromophenacyl bromide (BPB) and arachidonyltrifluoro-methyl ketone ($AACOCF_3$) also suppressed the neurite outgrowth of the cells, as well Taken together, these data indicated that $cPLA_2$ is involved in NGF-induced neuronal differentiation and neurite outgrowth of PC12 cells.

• Archives of Pharmacal Research
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• v.24 no.4
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• pp.307-311
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• 2001

The activation of NF-$\kappa$B induced by kojic Acid, an inhibitor of tyrosinase for biosynthesis of melanin in melanocytes, was investigated in human transfectant HaCaT and SCC-13 cells. These two keratinocyte cell lines transfected with pNF-$\kappa$B-SEAP-NPT plasmid were used to determine the activation of NF-$\kappa$B. Transfectant cells release the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the NF-$\kappa$B activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selective marker of geneticin resistance. NF-$\kappa$B activation was measured in the SEAP reporter gene assay using a fluorescence detection method. Kojic Acid showed the inhibition of cellular NF-$\kappa$B activity in both human keratinocyte transfectants. It could also downregulate the ultraviolet ray (UVR)-induced activation of NF-$\kappa$B expression in transfectant HaCaT cells. Moreover, the inhibitory activity of kojic Acid in transfectant HaCaT cells was found to be more potent than known antioxidants, e.g., vitamin C and N~acetyl-L-cysteine. These results indicate that kojic Acid is a potential inhibitor of NF-$\kappa$B activation in human keratinocytes, and suggest the hypothesis that NF-$\kappa$B activation may be involved in kojic Acid induced anti-melanogenic effect.

• YAKHAK HOEJI
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• v.43 no.6
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• pp.716-728
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• 1999

Whole oil and ketonic fraction (KF) of Leptospermum scoparium have been tested for their antimicrobial activity and combination effect with several antibiotics against various bacterial strains and fungi by using microbiological assay methods. Antibacterial activities of KF against a number of test strains were 2-3 fold stronger than those of whole oil. MICs of the KF were $65~125{\;}{\mu\textrm{g}}/ml$ against seven gram positive bacterial strains, $65~250{\;}{\mu\textrm{g}}/ml$ against 19 methicillin resistance Staphylococcus aureus strains, and $65~50{\;}{\mu\textrm{g}}/ml$ against 14 quinolone resistance strains. However, KF showed little or no activity against gram negative bacteria. MICs of the KF were $16~250{\;}{\mu\textrm{g}}/ml$ against more than 50% of the anaerobic bacterial strains tested. KF showed the higher antibacterial activity than bacitracin against 10 strains of Bacteroids thetaiotaomicron, or three strains of Bacteroides ovatus, and the more active than ciprofloxacin against one strain of Bacteroides thetaiotaomicron and three strains of Bacteroids ovatus. The MICs of KF was 63 and $250{\;}{\mu\textrm{g}}/ml$ against Aspergillus niger and Candida albicans, respectively. Antibacterial activities of KF in combination with 19 antibiotics against 14 strains and with four antifungal agents against one fungal strain were determined by paper strip diffusion method. While most of combination showed additivity, KF showed synergism with bacitracin, exfadroxil, cephradin, and meropenem for 29~57% of the strains tested. However, ofloxacin, enoxacin, sparfloxacin showed antagonism with KF for 43~71% of the strains. KF alone and in combination with bacitracin, gentamycin, neomycin, itraconazole, fluconazole, terfinafine and ketoconazole against five bacterial strains or one fungus strain synergistic effect was demonstrated against 33% of strains examined with FIC index value below 0.5 by checkerboard study. Synergistic effect of KF with gentamicin against Staphylococcus epidermidis 329 (QRS) was found by time-kill study.

• Food Science and Biotechnology
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• v.16 no.1
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• pp.99-103
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• 2007

The purpose of this study was to develop a simple detection method, possibly at the species-level, that allows for large-scale screening of bifidobacteria. Human fecal samples were plated on MRS-raffinose agar containing cysteine and neomycin sulfate, serving as selective pressure for bifidobacteria, and 0.003%(w/v) bromocresol green. All of the test strains grew well on this medium at $37{\pm}1^{\circ}C$, forming white colonies surrounded by yellow halos, which presented a sharp contrast against the green background. In this disc assay, the required incubation time to develop a yellowish zone varied with the species of Bifidobacterium that was tested, allowing for differential counts and easy identification at the species-level: 10-14 hr for B. bifidum, 20-22 hr for B. catenulatum and B. infantis. and 24-25 hr for B. longum and B. breve. No apparent color was observed for B. angulatum and B. adolescentis 28 hr after inoculation. To evaluate the results of pH indicator-based identification, individual isolates were subjected to a colony-PCR experiment with genus-specific primers. The amplified products from the isolates were in good accordance with those from the reference strains at a level of 95% agreement. These results suggest that the present method could be conveniently applied to cell counts, as well as to the preliminary identification of bifidobacteria from a variety of sample types including human feces, dairy products, and commercial probiotic supplements.

• Korean Journal of Veterinary Research
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• v.25 no.1
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• pp.53-59
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• 1985

A group of 80 Holstein calves, many with clinical signs of infectious bovine keratoconjunctivitis in summer seasons, was assembled for bacteriologic and ophthalmologic studies at an integrated dairy farm. Observations were started from 1980 to 1984 but bacteriological study was only carried out from thor spring in 1981 and continued during the Autumn in 1981. Corneal lesions and drugs sensitivity were also observed and the results obtained were as follows: 1. Occurrences of IBK-affected cattle during the 5 years were predominent to calves in summer. 2. Among 142 eyes from 80 cattle, 53 isolates of Moraxella bovis(37.3%) were isolated from 37 cattle(46.3%). The prevalence of the infection for clinical eyes were higher than nonclinical eyes calves. 3. Miscellaneous organisms were found from age groups of 5 months to 9 months old. It were 8 hemolytic Neisseria spp., 3 Fungi, Mycoplasma sp. and 1 Pseudomonas sp., but 9 samples were unknown. 4. Prevalence of infection with M. bovis in bilateral clinical eyes were higher than unilateral eyes. 5. Incidence of corneal lesion was predominent in early stage as a watery tears(21.0%) and late stage as a leukomas of corneal opacity(22.8%). 6. The number of isolations of M. bovis and incidence of IBK varied from year to year; Higher incidence of IBK clinical signs were showed in calves from 5 months to 10 months old and the number of isolations of M. bovis was declined in order 8 months(100.0%), 4 months(75.0%), 5 months(66.6%) and 8 months(66.6%) old calves. 7. Chloramphenicol, Nitrofurantoin and Cephalothin were highly sensitive against M. bovis isolates and then Tetracycline, Neomycin, Erythromycin and Kanamycin were intermediate but low sensitive to Streptomycin, Colistin and Penicillin.

• Korean Journal of Plant Tissue Culture
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• v.25 no.3
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• pp.201-206
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• 1998

To develop the fruits of cucumber (Cucumis sativus L.) producing high yields of superoxide dismutase (SOD), the MnSOD cDNA from pea (Pisum sativum) under the control of the cauliflower mosaic virus 35S promoter was introduced into cucumber using Agrobacterium tumefaciens (strain LBA 4404)-mediated transformation. The kanamycin-resistant shoots were selected on the selection medium containing MS basal salt, 1.0 mg/L zeatin, 0.1 mg/L IAA, 300 mg/L claforan, and 100 mg/L kanamycin. After 6 weeks of culture on the selection medium, the shoots were transferred to MS medium containing 0.2 mg/L NAA to induce roots. PCR analysis using the primers for neomycin phosphotransferase (NPTII) gene revealed that three plantlets were transformed. The fruits of one transgenic plant had approximately 3.2-fold higher SOD activity than those of non-transgenic plants. MnSOD isoenzyme band was strongly detected on native gel in fruits of transgenic plants.

• Food Science of Animal Resources
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• v.27 no.4
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• pp.509-516
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• 2007

Starters of lactic acid bacteria(LAB) were isolated from the commercial yoghurt products and the four isolates have been studied on their identification and some physiological characteristics. For the purpose of identification, microscopic examination, API test, and 16s rRNA gene sequencing were conducted. Isolate A from a yogurt product of local dairy company A was shown to be Gram-positive rod-shaped bacterium. All strains isolated were turned out to be as Lactobacillus paracasei by using a API 50 CHL kit. In contrast, isolate A was identified as a strain of Lactobacillus helveticus based on the 16S rRNA sequencing data, and L. casei ssp. casei for both B and D and L. paracasei for C. All the isolates survived the simulated gastric juice, pH 2.0 within 3 hours and sharply decreased in viability so that no viable cell was observed after 4.5 hours incubation. In addition, the four isolated strains were almost identical in antibiotic susceptibility to six different kinds of antibiotics including erythromycin ($15\;{\mu}g$), ampicillin ($10\;{\mu}g$), gentamycin ($10\;{\mu}g$), neomycin ($30\;{\mu}g$), but rather resistant to colistin ($10\;{\mu}g$) and streptomycin ($10\;{\mu}g$). It was noteworthy that four isolates were confirmed to produce antibacterial substance against foodborne pathogens of Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli 0157:H7 as test organisms based on the inhibitory zones on an MRS soft agar medium. At presence, the inhibitory factor is unknown so that further studies are required to ascertain the active factor responsible for the inhibitory activities.