• 치위생과학회지
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• 제20권4호
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• pp.213-220
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• 2020

Background: The leaves of Perilla frutescens, commonly called perilla and used for food in Korea, contain components with a variety of biological effects and potential therapeutic applications. The purpose of this study was to identify the components of 70% ethanol extracted Perilla frutescens (EEPF) and determine its inhibitory effects on oral microbial activity and production of nitric oxide (NO) and prostaglandin E2 (PGE2) in lipopolysaccharides (LPS)-stimulated Raw264.7 macrophages, consequently, to confirm the possibility of using EEPF as a functional component for improving the oral environment and preventing inflammation. Methods: One kg of P. frutescens leaves was extracted with 70% ethanol and dried at -70℃. EEPF was analyzed using high-performance liquid chromatography analysis, and antimicrobial activity against oral microorganisms was revealed using the disk diffusion test. Cell viability was elucidated using a methylthiazolydiphenyl-tetrazolium bromide assay, and the effect of EEPF on LPS-induced morphological variation was confirmed through microscopic observation. The effect of EEPF on LPS-induced production of pro-inflammatory mediators, NO and PGE2 was confirmed by the NO assay and PGE2 enzyme-linked immunosorbent assay. Results: The main component of EEPF was rosemarinic acid, and EEPF showed weak anti-bacterial and anti-fungal effects against microorganisms living in the oral cavity. EEPF did not show toxicity to Raw264.7 macrophages and had inhibitory effects on the morphological variations and production of pro-inflammatory mediators, NO and PGE2 in LPS-stimulated Raw264.7 macrophages. Conclusion: EEPF can be used as a functional material for improving the oral environment through the control of oral microorganisms and for modulating inflammation by inhibiting the production of inflammatory mediators.

• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.989-1002
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• 2022

Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

• 한국미생물·생명공학회지
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• 제51권1호
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• pp.10-17
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• 2023

Dairy products are extensively used as carriers of probiotic strains that have potential health benefits. Assessment of the viability of probiotic strains during manufacturing is important to ensure that products meet recommended levels. Hence, the method for accurately quantifying lactic acid bacteria (LAB) in probiotic or dairy products is required. The present study aims to examine the performance of de-Man Rogosa Sharpe (MRS), plate count agar with bromocresol purple (PCA with BCP), and glucose blood liver (BL) agars recommended in the Korea Food Code guidelines for counting LAB. Analysis of the performance of culture media containing 19 lactic acid bacterial species commonly encountered in probiotic and dairy products showed no statistically significant difference between 18 reference strains and three culture media (p > 0.01). Furthermore, the suitability of three culture media was verified for the quantitative assessment of LAB in 25 probiotic and dairy products. The number of LAB in three culture media was determined to be more than 10⁷ colony-forming unit (CFU)/ml for fermented milk products and 10⁸ CFU/ml for condensed fermented milk and probiotic products, indicating that they all satisfied the Korea Food Code guidelines. Moreover, there was no statistically significant difference in the amount of LAB counted in all three culture media, suggesting that they can be used to isolate or enumerate LAB in commercial products. Finally, three culture media will be useful for isolating and enumerating LAB from fermented foods as well as gut microflora.

• 환경생물
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• 제41권2호
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• pp.167-178
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• 2023

Large amounts of waste and wastewater from aquaculture have negatively impacted ecosystems. Among them, shrimp aquaculture wastewater contains large amounts of nitrogen contaminants derived from feed residues in an aerobic environment. This study isolated candidate strains from adult rockworms to treat shrimp aquaculture wastewater (SAW) in an aerobic environment. Among 87 strains isolated, 25 grew well at the same temperature as the shrimp aquaculture with excellent polymer degradation ability (>0.5 cm clear zone). Six isolates (strains AL1, AL4, AL5, AL6, LA10, and PR15) were finally selected after combining strains with excellent polymer degradation ability without antagonism. 16S rRNA sequencing analysis revealed that strains AL1, AL4, AL5, AL6, LA10, and PR15 were closely related to Bacillus paramycoides, Bacillus pumilus, Stenotrophomonas rhizophila, Bacillus paranthracis, Bacillus paranthracis, and Micrococcus luteus, respectively. When these six isolates were applied to SAW, they reached a maximum cell viability of 2.06×10⁵ CFU mL^-1. Their chemical oxygen demand (COD_Cr) and total nitrogen(TN) removal rates for 12h were 51.0% and 44.6%, respectively, when the COD_Cr/TN ratio was approximately 10.0. Considering these removal rates achieved in this study under batch conditions, these six isolates could be used for aerobic denitrification. Consequently, these six isolates from rockworms are good candidates that can be applied to the field of aquaculture wastewater treatment.

• 대한본초학회지
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• 제24권2호
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• pp.21-27
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• 2009

Objectives: Spatholobi Caulis is the stem of Spatholobus suberectus Dunn., which has been used for treating menstrual disorders, anemia, and rheumatic arthralgia in traditional Oriental medicine, This study was conducted to evaluate the inhibitory effects of Spatholobi Caulis extract (SCE) on the production of nitric oxide (NO) and pro-inflammatory cytokines in LPS-activated Raw264,7 cells. Methods : Cell viability was determined by MIT assay. Relative levels of NO were measured with Griess reagent and pro-inflammatory cytokines were detected by ELISA Expression of iNOS and COX-2 proteins were determined by western blotting. Results : NO production and iNOS expression were increased by incubation with LPS for 24 h. However, the augmented NO was reduced by SCE in a dose-dependent manner. Expressions of iNOS and COX-2 were inhibited by the treatment with SCE, SCE also suppressed the production of pro-inflammatory cytokines, such as IL-1beta and IL-6. Conclusions : These results indicate that the MeOH extract of Spatholobi Caulis warrant further development as an anti-inflammatory agent for the treatment of Gram-negative bacterial infections.

• Restorative Dentistry and Endodontics
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• 제46권2호
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• pp.20.1-20.11
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• 2021

Objectives: The aim of this study was to evaluate bacterial nanocellulose (BNC) membranes incorporated with antimicrobial agents regarding cytotoxicity in fibroblasts of the periodontal ligament (PDLF), antimicrobial activity, and inhibition of multispecies biofilm formation. Materials and Methods: The tested BNC membranes were BNC + 1% clindamycin (BNC/CLI); BNC + 0.12% chlorhexidine (BNC/CHX); BNC + nitric oxide (BNC/NO); and conventional BNC (BNC; control). After PDLF culture, the BNC membranes were positioned in the wells and maintained for 24 hours. Cell viability was then evaluated using the MTS calorimetric test. Antimicrobial activity against Enterococcus faecalis, Actinomyces naeslundii, and Streptococcus sanguinis (S. sanguinis) was evaluated using the agar diffusion test. To assess the antibiofilm activity, BNC membranes were exposed for 24 hours to the mixed culture. After sonicating the BNC membranes to remove the remaining biofilm and plating the suspension on agar, the number of colony-forming units (CFU)/mL was determined. Data were analyzed by 1-way analysis of variance and the Tukey, Kruskal-Wallis, and Dunn tests (α = 5%). Results: PDLF metabolic activity after contact with BNC/CHX, BNC/CLI, and BNC/NO was 35%, 61% and 97%, respectively, compared to BNC. BNC/NO showed biocompatibility similar to that of BNC (p = 0.78). BNC/CLI showed the largest inhibition halos, and was superior to the other BNC membranes against S. sanguinis (p < 0.05). The experimental BNC membranes inhibited biofilm formation, with about a 3-fold log CFU reduction compared to BNC (p < 0.05). Conclusions: BNC/NO showed excellent biocompatibility and inhibited multispecies biofilm formation, similarly to BNC/CLI and BNC/CHX.

• Journal of Oral Medicine and Pain
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• 제32권2호
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• pp.137-150
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• 2007

피톤치드란 '산림향' 이라고 부르는, 나무가 갖는 특유의 향을 발산하는 휘발성 화학물질로서, 우리 몸을 쾌적하게 해 줄 뿐만 아니라 항균, 방충, 소취 등 다양한 기능을 가지고 있다. 치주질환과 구취를 유발시키는 중요한 원인균인 P. gingivalis에 대한 피톤치드의 항균효과와 항균작용을 연구하기 위하여, 편백 피톤치드와 함께 P. gingivalis 2561을 배양한 후 P. gingivalis 2561의 성장정도, 생존력 및 형태적, 분자생물학적 변화를 관찰하여 다음과 같은 결과를 얻었다. 1. 피톤치드는 P. gingivalis에 매우 강한 항균력을 보였고, 이 항균력은 살균작용에 의한 것으로 나타났다. P. gingivalis에 대한 피톤치드의 최소억제농도는 0.008%, 최소살균농도는 0.01%로 결정되었다. 2. 피톤치드와 함께 배양된 P. gingivalis는 ampicillin, cefatoxime, penicillin, tetracycline에 대한 감수성이 변하지 않았으나 amoxicillin에 대한 감수성은 증가하였다. 3. 피톤치드와 같이 배양된 P. gingivalis를 투과전자현미경으로 관찰한 결과, 핵이 뚜렷해지고 전자밀도가 높은 과립이 증가하였고 리보솜이 세포질 가장 자리로 분포하였으며, 피톤치드 양이 증가할 수록 유령세포, 특히 소포가 특징적으로 크게 증가하였다. 4. RT-PCR 분석 결과, 피톤치드는 P. gingivalis의 superoxide dismutase의 발현을 억제하는 것으로 나타났다. 5. SDS-PAGE와 immunoblot 분석 결과, 피톤치드는 P. gingivalis의 단백질 발현에 큰 영향을 미치지 않는 것으로 나타났다. 이상의 결과로 미루어 피톤치드는 P. gingivalis에 대해 강한 항균효과를 갖고 있으며 이것은 살균작용에 의한 것으로 판단된다. 즉, 피톤치드는 단백질 발현에는 영향을 미치지 못하지만 P. gingivalis의 항산화물질 생산능력을 감소시키거나 아직 밝혀지지 않은 기전을 통해 스트레스 상황을 유도하여 생존능력을 억제하여 결과적으로 세균세포의 구조적 형태 변화와 함께 사멸을 유도하는 것으로 사료된다.

• 대한소아치과학회지
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• 제40권4호
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• pp.241-246
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• 2013

이 연구의 목적은 치과 임상에서 널리 사용되는 항미생물제제인 클로르헥시딘과 할로겐을 이용한 erythrosine 광역동 치료를 병용하였을 때, 부유 상태의 Streptococcus mutans에 미치는 항균효과를 평가하기 위함이다. S. mutans 표준 균주를 Brain Heart Infusion broth medium에 첨가하여 부유 상태로 배양하였다. 치과용 할로겐 광중합 장치를 광원으로 사용하였으며 치태 착색제인 erythrosine을 광감각제로 사용하였다. 클로르헥시딘의 최소살균농도를 측정하고, erythrosine 광역동 치료에서 최소살균농도를 결정하기 위해 $1{\mu}M$, $5{\mu}M$, $10{\mu}M$의 erythrosine 농도를 사용하여 할로겐 광중합기로 15초 동안 광조사를 시행한 후 세균수를 측정하였다. 마지막으로 클로르헥시딘과 erythrosine 광역동 치료를 병용한 경우 나타나는 상승효과를 평가하기 위해, 최소살균농도 이하인 0.001% 클로르헥시딘과 최소살균농도 이하인 $1{\mu}M$ erythrosine 광역동 치료를 병용하여 항균효과를 비교하였다. 최소살균농도 이하의 클로르헥시딘만 단독으로 사용하는 경우 S. mutans의 세균수가 유의하게 감소하지 않았다(p < 0.05). 또한 erythrosine 광역동 치료만 단독으로 시행한 경우에는 $5{\mu}M$, $10{\mu}M$의 erythrosine 농도에서 유의하게 세균수가 감소하였지만, $1{\mu}M$ erythrosine 농도에서는 세균수가 유의하게 감소하지 않았다(p < 0.05). 마지막으로 두 가지 치료법의 상승효과를 평가하기 위해 최소살균농도 이하인 $1{\mu}M$ erythrosine 광역동 치료와 0.001% 클로르헥시딘을 병용하는 경우에는, 대조군과 비교하여 세균수가 유의하게 감소하였다(p < 0.05). 최소살균농도 이하인 0.001% 클로르헥시딘과 $1{\mu}M$ erythrosine 광역동 치료를 각각 단독으로 시행하는 경우에는 유의한 항균효과가 없었지만, 두 가지 치료를 병용하는 경우 61%의 S. mutans 세균수가 감소하였다. 이상의 결과를 종합해 볼때 S. mutans에 대해 최소살균농도 이하의 클로르헥시딘과 erythrosine 광역동 치료를 병용하는 경우 상승효과가 있음을 알 수 있다.

• 농업생명과학연구
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• 제44권6호
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• pp.111-119
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• 2010

살모넬라증은 대표적인 인수공통전염병의 하나로서 세포내 기생하며 질병을 유발하며 장염과 식중독 등을 유발하여 공중보건학적으로 심각한 문제를 야기하고 있다. 본 연구는 삼백초의 수용성 추출물을 (SCWE) 이용하여 숙주세포에 대한 안전성, S. typhimurium 균에 대한 항균효과 및 대식세포 내 균 증식억제 기능을 규명하였다. 본 실험을 통하여 SCWE 1, 10 및 $100{\mu}$g/ml 농도로 첨가한 배지에서 RAW 264.7 체포와 24 시간 반응 후 평가해본 결과 세포독성이 인정되지 않았으며, S. typhimurium 균에 대하여 시간 경과에 따라 항균효과가 증가되는 것을 확인하였고, SCWE 처리에 의해 대식세포의 형태적 변화가 증가되는 것으로 나타났으며 (p<0.05), 살모넬라균의 탐식능력 및 대식세포 내 균 증식 억제 능력이 비 처리군에 비해 현저히 증가되는 것이 확인되었다. 또한 SCWE 처리 한 대식세포에 살모넬라균 감염을 수행하였을 때 대식세포의 nitric oxide (NO) 산생능력이 비 처리 군에 비해 저하되는 것으로 나타나, 살모넬라균에 의한 대식세포의 세포독성을 억제하는 것으로 나타났다. 종합적으로 SCWE의 숙주세포에 대한 안전성, 살모넬라균에 대한 항균효과 및 대식세포 내 균 증식 억제 효과가 있는 것으로 나타나 SCWE를 이용한 세포내 기생세균의 치료제 개발이 가능할 것으로 판단된다.

• 대한한의학방제학회지
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• 제15권1호
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• pp.163-173
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• 2007

Jeungaektang (JAT) is the herbal formula, has the effect of moistening the dryness by activating lung Qi and by nourishing Yin, has being used for dryness syndromes. Generally the herbal formulae for moistening dryness are used for exogenous or endogenous dryness syndromes. JAT has been clinically used for the treatment of endogenous dryness syndromes. It is composed of Scrophulariae Radix. Rehmanniae Radix and Liriopis Tuber. Recent studies showed that JAT has a protective effect against $CCl_{4}-induced$ hepatotoxicity and anti-inflammatory effects against ear swelling of mouse induced by Crotonis Fructus. However, the effect of JAT on the immunological activity was rarely studied. Therefore, this study evaluated the effects of JAT the regulatory mechanism of nitric oxide (NO) and cytokines in the lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. After the treatment of JAT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX -2) and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidence that JAT inhibited the production of nitrite and nitrate ($0.1{\sim}1.0$ mg/ml), iNOS ($0.1{\sim}1.0$ mg/ml), $interleukin-1{\beta}$ ( $0.1{\sim}1.0$ mg/ml) and tumor necrosis $factor-{\alpha}$ ($0.1{\sim}1.0$ mg/ml) in RAW 264.7 cells activated with LPS. Furthermore, JAT inhibited the expression of COX-2 expression and production of prostagladin E2 ($0.1{\sim}1.0$ mg/ml). These findings suggest that JAT can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.