• Korean Journal of Veterinary Research
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• v.29 no.3
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• pp.393-405
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• 1989

This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

• Weed & Turfgrass Science
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• v.4 no.2
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• pp.118-123
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• 2015

Metagenomics is a powerful tool to isolate novel biocatalyst and biomolecules directly from the environmental DNA libraries. Since the metagenomics approach bypasses cultivation of microorganisms, un-cultured microorganisms that are majority of exists can be the richest reservoir for natural products discovery. To discover novel herbicidal substances from soil metagenome, we established three easy, simple and effective high throughput screening methods such as cucumber cotyledon leaf disc assay, microalgae assay and seed germination assay. Employing the methods, we isolated two active single clones (9-G1 and 9-G12) expressing herbicidal activity which whitened leaf discs, inhibited growth of microalgae and inhibited root growth of germinated Arabidopsis seeds. Spraying butanol fraction of the isolated active clones' culture broth led to growth retardation or desiccation of Digitalia sanguinalis (L) Scop. in vivo. These results represent that the screening methods established in this study are useful to screen herbicidal substances from metagenome libraries. Further identifying molecular structure of the herbicidal active substances and analyzing gene clusters encoding synthesis systems for the active substances are in progress.

• Restorative Dentistry and Endodontics
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• v.35 no.5
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• pp.359-367
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• 2010

Objectives: The purpose of the present in vitro study was to evaluate the biocompatibility of mineral trioxide aggregate (MTA) mixed with glass ionomer cement (GIC), and to compare it with that of MTA, GIC, IRM and SuperEBA. Materials and Methods: Experimental groups were divided into 3 groups such as 1 : 1, 2 : 1, and 1 : 2 groups depending on the mixing ratios of MTA powder and GIC powder. Instead of distilled water, GIC liquid was mixed with the powder. This study was carried out using MG-63 cells derived from human osteosarcoma. They were incubated for 1 day on the surfaces of disc samples and examined by scanning electron microscopy. To evaluate the cytotoxicity of test materials quantitatively, XTT assay was used. The cells were exposed to the extracts and incubated. Cell viability was recorded by measuring the optical density of each test well in reference to controls. Results: The SEM revealed that elongated, dense, and almost confluent cells were observed in the cultures of MTA mixed with GIC, MTA and GIC. On the contrary, cells on the surface of IRM or SuperEBA were round in shape. In XTT assay, cell viability of MTA mixed with GIC group was similar to that of MTA or GIC at all time points. IRM and SuperEBA showed significantly lower cell viability than other groups at all time points (p < 0.05). Conclusions: In this research MTA mixed with GIC showed similar cellular responses as MTA and GIC. It suggests that MTA mixed with GIC has good biocompatibility like MTA and GIC.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.51-58
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• 2015

Objectives : This paper aimed to verify the applicability of mixed extract ofAngelica gigasNakai,Cnidium officinaleMakino,Paeoniala ctifloraPall,Rechmannia glutinosaLibosch,Scutellaria baicalensisGeorgi, which were prescribed for improving inflammation in Donguibogam, as the materials for beauty food and functional medicinal herb cosmetics by manufacturing such mixed extract and evaluating the biological activity of the extract.Methods : The mixed medicinal herb water extract(MMW) and ethanol extract(MME) were freeze-dried to be used as the specimen. We performed electron donating ability, lipid acidification inhibitory activity, anti-inflammatory activity against skin flora, MTT assay, NO inhibitory activity and the protein expression inhibitory activity of iNOS and COX-2.Results : For anti-oxidation experimentation, the electron donating abilities of MMW and MME were above 60.0% and 90.0% at 500 μg/ml, respectively. In the inhibition rate of lipid peroxidation, MMW and MME showed 43.1% and 52.1% at 1,000 μg/ml, respectively. As a result of antimicrobial activity, both the MMW and MME showed significant clear zones forPropionibacterium acnesat 4 mg/disc, but did not indicated the clearzones forStaphylococcus aureus, Escherichia coliandStaphylococcus epidermidis. Anti-inflammatory activity by NO assay showed LPS-induced NO was significantly inhibited in a concentration-dependent manner. Also, the expression of iNOS and COX-2 proteins were significantly inhibited following treatment with MMW and MME of 50 μg/ml.Conclusions : Accordingly, it can be concluded that mixed medicinal herb extract has the potential to beused as a functional food and cosmetic material.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.15-21
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• 2015

Objectives : Methicillin-resistantStaphylococcus aureus(MRSA) is a human pathogen. New antibacterial agents are needed to treat MRSA-related infections. This study investigated the antibacterial activity of EtOH 70% extracts ofTonghyeonipal-dan(THD) which prescription is composed of oriental medicine against MRSA.Methods : The antibacterial activity of THD was evaluated against MRSA strains by using the Disc diffusion method, broth microdilution method, Checkerboard dilution test, and Time-kill test; its mechanism of action was investigated by bacteriolysis, detergent or ATPase inhibitors were used.Results : The minimum inhibitory concentration (MIC) of THD is 1,000~2,000 μg/mL against MRSA. In the checkerboard dilution test, fractional inhibitory concentration index (FICI) of THD in combination with antibiotics indicated synergy or partial synergism againstS. aureus. Furthermore, a time-kill assay showed that the growth of the tasted bacteria was considerably inhibited after 24 h of treatment with the combination of THD with selected antibiotics. For measurement of cell membrane permeability, THD 500 μg/mL along with concentration of Triton X-100 (TX) and Tris-(hydroxymethyl) aminomethane (TRIS) were used. In the other hand, N,N-dicyclohexylcarbodimide (DCCD) and Sodium azide (NaN3) were used as an inhibitor of ATPase. TX, TRIS, DCCD and NaN3 cooperation againstS. aureusshowed synergistic action.Conclusions : Accordingly, antimicrobial activity of THD was affected by cell membrane and inhibitor of ATPase were assessed. These results suggest that THD has antibacterial activity, and that THD extract offers great potential as a natural antibiotic against MRSA.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.187-194
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• 2015

In this study, L. plantarum, when reacting with the culture media of potential pathogenic bacteria, exhibited an increase in growth rate and antimicrobial activity. In order to examine the characteristics and the nature of the reaction with the bacteria, this study carried out experiments involving culturing the test bacteria in M9 minimal media. Subsequently, the supernatant was incrassated by the decompression-drying method. Through colony forming unit assay, it was confirmed that L. plantarum had the function of growth inhibition to various bacteria. After culturing L. plantarum with bacterial media, the growth rate of L. plantarum was measured by absorbance (OD₆₀₀), the results showed that the growth rate (E. coli treatment group: OD₆₀₀ = 0.848, S. typhimurium treatment group: OD₆₀₀ = 0.848) increased, as compared with the non-treated control group (OD₆₀₀ = 0.48). In contrast, the concentrate itself did not induce the growth of L. plantarum. These results were observed as a universal phenomenon of the Lactobacillus species. Moreover, the increase in antimicrobial activity was observed in L. plantarum, which reacted with the culture media of E. coli and S. typhimurium, through a disc diffusion assay, and the result of growth inhibition against various bacteria was induced. Finally, based on the analysis results of the characteristics of bacteria culture media, which increased the growth rate of L. plantarum and antibacterial activity, the bacterial media had a tolerance for catabolic enzymes, pH 2−8 and heat. Therefore, this substance can be said to be a small molecule which is highly stable under various conditions.

• The Journal of Advanced Prosthodontics
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• v.8 no.3
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• pp.172-180
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• 2016

PURPOSE. The purpose of this study was to analyze the antimicrobial, antioxidant activity and cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract for assessing whether Dendropanax morbifera $L{\acute{e}}veille$ can be used for the development of natural mouthwash and denture cleaning solution. MATERIALS AND METHODS. The extract was obtained from branches of Dendropanax morbifera $L{\acute{e}}veille$. The solvent fractions were acquired by fractionating Dendropanax morbifera $L{\acute{e}}veille$ extract using n-hexane, ethyl acetate, chloroform and butanol solvent. Paper disc test was used to evaluate the antimicrobial and antifungal activity of Dendropanax morbifera $L{\acute{e}}veille$ extract and solvent fractions against Streptococcus mutans and Candida albicans. The analysis of antioxidant activity was carried out through DPPH radical scavenging assay. The cytotoxicity of Dendropanax morbifera $L{\acute{e}}veille$ extract was analyzed through MTT assay using normal human oral keratinocytes. RESULTS. Dendropanax morbifera $L{\acute{e}}veille$ extract showed antimicrobial activity against Streptococcus mutans and especially Candida albicans. The solvent fractions of Dendropanax morbifera $L{\acute{e}}veille$ showed strong antimicrobial activity against Streptococcus mutans and Candida albicans in n-hexane and butanol solvent fraction, respectively. Dendropanax morbifera $L{\acute{e}}veille$ extract also showed outstanding antioxidant activity. Butanol, ethyl acetate, and chloroform solvent fraction of Dendropanax morbifera $L{\acute{e}}veille$ tended to have increased antioxidant activity as the concentration increased. Dendropanax morbifera $L{\acute{e}}veille$ extract showed high cell survival rate in cytotoxicity test. CONCLUSION. Dendropanax morbifera $L{\acute{e}}veille$ extract turned out to have antimicrobial, antioxidant activity and cytophilicity. Based on these results, it is expected that Dendropanax morbifera $L{\acute{e}}veille$ is applicable as an ingredient for natural mouthwash and denture cleanser.

• Journal of Pharmacopuncture
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• v.9 no.3 s.21
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• pp.97-104
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• 2006

Objectives : This study was conducted to compare antibacterial activities and free radical scavenging activity between the Bee Venom and Sweet Bee Venom in which the allergy-causing enzyme is removed. Methods : To evaluate antibacterial activities of the test samples, gram negative E. coli and gram positive St. aureus were compared using the paper disc method. For comparison of the antioxidant effects, DPPH(1,1-diphenyl-2picrylhydrazyl) free radical scavenging assay and Thiobarbituric Acid Reactive Substances(TBARS) assay were conducted. Results : 1. Antibacterial activity against gram negative E. coli was greater in the Sweet Bee Venom group than the Bee Venom group. 2. Antibacterial activity against gram positive St. aureus was similar between the Bee Venom and Sweet Bee Venom groups. 3. DPPH free radical scavenging activity of the Bee Venom group showed 2.8 times stronger than that of the Sweet Bee Venom group. 4. Inhibition of lipid peroxidation of the Bee Venom group showed 782 times greater than that of the Sweet Bee Venom group. Conclusions : The Bee Venom group showed outstanding antibacterial activity against gram positive St. aureus, and allergen-removed Sweet Bee Venom group showed outstanding antibacterial activity against both gram negative E. coli and gram positive St. aureus. For antioxidant effects, the Bee Venom was superior over the Sweet Bee Venom and the superiority was far more apparent for lipid peroxidation.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.26-35
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• 2017

The hydrolysis of proteins constitutes an invaluable tool, granting access to a variety of peptide fragments with potentially interesting biological properties. Therefore, a hemoglobin (Hb) hydrolysate of Crocodylus siamensis was generated by digestion under acidic conditions. The antibacterial and antioxidant activities of the Hb hydrolysate were assessed in comparison with intact Hb. A disc diffusion assay revealed that the Hb hydrolysate exhibited antibacterial activity against eight strains of gram-positive bacteria and showed a higher efficacy than intact Hb. Moreover, the antioxidant activity of intact Hb and its hydrolysate was evaluated using ABTS and DPPH radical scavenging assays. The Hb hydrolysate exhibited free radical scavenging rates of 6-32%, whereas intact Hb showed a slightly higher activity. In addition, non-toxicity to human erythrocytes was observed after treatment with quantities of Hb hydrolysate up to $10{\mu}g$. Moreover, active fragmented Hb (P3) was obtained after purifying the Hb hydrolysate by reversed-phase HPLC. Scanning electron microscopy demonstrated the induction of bacterial cell membrane abnormalities after exposure to P3. Antibacterial and antioxidant activities play crucial roles for supporting the wound healing activity. Consequently, an in vivo mice excisional skin wound healing assay was carried out to investigate the effects of intact Hb treatment on wound healing in more detail. The results clearly demonstrate that intact Hb is capable of promoting 75% wound closure within 6 days. These findings imply that intact Hb of C. siamensis and its acid hydrolysate may serve as valuable precursors for food supplementary products benefitting human health.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.10a
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• pp.91-91
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• 2018

The Rosa multiflora, a well-known plant belonging to Rosacea, is widely used in orthodox medicine in worldwide. However, its biological activity as a functional ingredient for cosmetic products have not yet been studied. Accordingly, an investigation of the above mentioned atrributes was performed on a 50% ethanol extract of Rosa multiflora. The antioxidant activities were determined by DPPH. Additionally, the contents of total phenols and flavonoids were analyzed. Also, the phenolic compounds were detected using HPLC. The melanogenesis regulatory effect was evaluated using melanin content and cellular tyrosinase activity in B16F10 melanoma cells. The elastase inhibitory activity assay was performed for anti-wrinkle effect. The antimicrobial activity was assessed using the disc diffusion assay. The DPPH radical scavenging ability, denoted by the $SC_{50}$ value was found to be $123.1{\mu}g/ml$, whereas that of positive control (ascorbic acid) was $27.5{\mu}g/mL$. The content of total polyphenol and flavonoid content were 202 mg/g and 86.77 mg/g, respectively. In addition, astragalin and gallic acid were identified in the extract. Also, the ethanol extract significantly inhibited ${\alpha}$-MSH-induced melanogenesis in B16F10 cells. For anti-wrinkle effect, elastase inhibition activity of the ethanol extract was 53.2% at a concentration of $100{\mu}g/ml$. The antimicrobial activity of the extract against S. aureus and E. coli was observed to be 0.5 - 5%, and no significant activity was noted against C. albicans. Therefore, the ethanol extract of Rosa multiflora can be used effectively for development of functional cosmetic materials.