• 한방비만학회지
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• 제19권2호
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• pp.106-112
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• 2019

Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4^th day and insulin medium at 6^th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×10⁷ CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

• 생명과학회지
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• 제23권3호
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• pp.406-414
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• 2013

강의 하류지역에서 미생물의 군집이 점진적인 염도의 증가에 따라 변한다는 것을 실험적으로 보기 위하여, 경북 형산강의 하류에서부터 연안해역으로 유입되는 곳까지 약 2.91 km 간격으로 0.02%, 1.48%, 2.63%, 3.62%의 염분을 포함하는 물 시료를 얻어 denaturing gradient gel electrophoresis (DGGE)를 수행하였다. 계통분석, 계통수 및 각 시료간의 연관성을 조사한 결과, 미생물 군집의 변화가 염분의 증가에 따라 점진적으로 변화하는 것을 확인하였으며, 이는 염분의 농도가 미생물 군집에 큰 영향을 미치는 요소임을 제시한다. 덧붙여 연안 지역이나 다른 수계환경에 비해 하류지역은 염분의 점진적인 변화로 인해 좁은 면적에 비하여 미생물 다양성이 크고, 이는 곧 특이하고 새로운 종을 찾기에 좋은 장소임을 시사하였다.

• 생명과학회지
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• 제23권10호
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• pp.1295-1303
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• 2013

현재까지 밝혀진 사실에 의하면 지구상에는 55개 phylum의 bacteria와 13개 phylum의 archaea가 존재하여 총 68개 phylum 있으며, 대장에는 9개 phylum의 미생물이 존재하여 일반 토양환경이나 해양환경에 비하여 상대적으로 낮은 미생물 다양성을 가지고 있다. 장내 미생물의 다양성은 Host의 고유특성(genetic background, sex, age, immune system, gut motility)과 식이(nondigestible carbohydrates fat, prebiotics or probiotics), 항생제 섭취 등으로 인하여 영향 받으며, 이러한 영향이 에너지 저장 과정과 유전자 발현을 영향을 주고 나아가 비만과 같은 대사질환에 영향을 미친다. Probiotics는 숙주의 장내 균총을 개선하여 건강에 유익함을 줄 수 있는 살아있는 미생물을 말하며, 주로 lactic acid bacteria들이 이용되어 왔다. 최근 probiotics를 이용한 설사나 과민성대장증후군 등과 같은 질병 치료를 위하여 bacteriotherapy가 활용되고 있다. Prebiotics는 장내 미생물을 선택적으로 조절할 수 있는 식품 성분으로 inulin, fructo-oligosaccharides, galacto-oligosaccharides, lactulose 등이 이용되고 있으며, 최근에는 비만, 항암과 관련하여 대장 내 식이섬유소의 이용과 단쇄지방산(short-chain fatty acids) 생산에 대한 관심이 집중되고 있다. 다양한 nondigestible carbohydrates와 prebiotics, probiotics에 의해서 조절되는 대장 내 특정 미생물 종을 규명하고, 이들 미생물 종이 생산하는 단쇄지방산과 같은 대사산물들과 질병과의 연관성 규명하는 대장 미생물 연구가 더욱 필요하다.

• Journal of Periodontal and Implant Science
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• 제38권2호
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• pp.143-152
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• 2008

Purpose: Specific bacteria are believed to play an important role in chronic periodontitis. Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systemic analysis of subingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to investigate the prevalence of 29 putative periodontal pathogens in Korean chronic periodontitis patients and evaluate which pathogens are more associated with Korean chronic periodontitis. Material and Methods: A total of 86 subgingival plaque samples were taken from 15 chronic periodontits(CP) patients and 13 periodontally healthy subjects in Korea. CP samples were obtained from the deepest periodontal pocket (>3 mm probing depth[PD]) and the most shallow periodontal probing site ($\leq$3 mm PD) in anterior tooth and posterior tooth, respectively, of each patient. Samples in healthy subjects were obtained from 1 anterior tooth and 1 posterior tooth. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed. Detection frequencies(% prevalence) of 29 putative periodontal pathogens were investigated as bacterium-positive sites/total sites. Results: With the exception of Olsenella profuse and Prevotella nigrescens, the sites of diseased patients generally showed higher prevalence than the healthy sites of healthy subjects for all bacteria analyzed. Tanerella forsythensis (B.forsythus), Campylobacter rectus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis and Porphyromonas gingivalis were detected in more than 80% of sites with deep probing depths in CP patients. In comparison between the sites (deep or shallow PD) of CP patients and the healthy sites of healthy subjects, there was statistically significant difference(P<0.05) of prevalence in T.forsythensis (B.forsythus), C.rectus, Dialister invisus, F.alocis, P.gingivalis and Treponema denticola. Conclusion: Our results demonstrate that the four putative periodontal pathogens, T.forsythensis (B.forsythus), C.rectus, P.gingivalis and F.alocis are closely related with CP patients in the Korean population.

• 한방비만학회지
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• 제20권1호
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• pp.40-51
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• 2020

Objectives: We aimed to compare the quality of sleep, tongue diagnosis, oral microbiology differences in insomnia of Liver qi stagnation (LQS) and Non-Liver qi stagnation (NLQS). Methods: 56 patients were classified as LQS or NLSQ type insomnia through the insomnia differentiation questionnaire. The depression scores between the groups were compared through beck depression inventory (BDI), and the sleep quality was compared through Pittsburgh sleep quality index (PSQI) and Insomnia Severity Index (ISI). We analyzed the sleep efficiency, total sleep time, total awake frequency, total and average awake time through actigraph. For the tongue diagnosis, the distribution of tongue coating in six areas were measured through Winkel tongue coating index (WTCI). Linear discriminant analysis was performed to observe the differences in composition of microbial strains between the groups. Results: The scores of BDI, ISI and PSQI were significantly higher in LQS group. The total sleep time in LQS group was significantly less than that of NLQS group. Among the areas of tongue, according to the WTCI, the amount of tongue coating in zones A and C was significantly small. In oral microbial analysis, there was no significant difference between the groups at the phylum level. At the genus level, Prevotella, Veillonella, and Streptococcus were predominant in LQS group, whereas Prevotella, Neisseria, and Streptococcus in NLQS group. Conclusions: It was meaningful that insomnia was more likely in LQS group than in NLQS group, and the composition of oral microorganisms was significantly different, which could lead to the diseases caused by stress.

• Journal of Microbiology and Biotechnology
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• 제30권9호
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• pp.1343-1354
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• 2020

Atopic dermatitis (AD) is a skin disorder that causes chronic itch. We investigated the inhibitory effects of a mixture of prebiotic short-chain galacto-oligosaccharides and long-chain fructooligosaccharides (scGOS/lcFOS), inulin, or β-glucan on AD development in 1-chloro-2,4-dinitrobenzene (DNCB)-treated NC/Nga mice. Mice were randomly assigned to six groups: untreated mice, AD control, positive control (DNCB-treated NC/Nga mice fed a dietary supplement of Zyrtec), and DNCB-treated NC/Nga mice fed a dietary supplement of prebiotics such as scGOS/lcFOS (T1), inulin (T2), or β-glucan (T3). The prebiotic treatment groups (T1, T2, and T3) showed suppression of AD symptoms, Th2 cell differentiation, and AD-like skin lesions induced by DNCB. In addition, prebiotic treatment also reduced the number of microorganisms such as Firmicutes, which is associated with AD symptoms, and increased the levels of Bacteroidetes and Ruminococcaceae, which are associated with alleviation of AD symptoms. Our findings demonstrate the inhibitory effects of prebiotics on AD development by improving the Th1/Th2 cytokine balance and beneficial symbiotic microorganisms in in vitro and in vivo models.

• Journal of Dairy Science and Biotechnology
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• 제32권2호
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• pp.141-145
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• 2014

시장에는 프로바이오틱스를 포함한 무수히 많은 제품이 판매되고 있는데, 우리에게 어떤 이로운 점을 줄 수 있는지 궁금해 한 적이 있었을 것이다. 프로바이오틱스는 기본적인 영양 측면을 배제하고, 우리가 유효한 양을 섭취하였을 때 건강상의 이점을 주는 살아있는 미생물로 정의된다. 프로바이오틱스는 유용하고 건강에 좋은 미생물로도 불려지고 있으며, 다음과 같은 다섯 가지 측면에서 건강 기능성이 있는 것으로 강조되어 왔는데, 1) 대장암 및 IBS와 같은 다른 대장(결장)관련 질병의 발병율 감소, 2) 면역 시스템의 촉진, 3) 항고혈압 및 항콜레스테롤 작용, 4) 장내세균에 작용하는 항생제의 효과를 경감시키고, 5) 위장관 감염을 예방시키는 점이다. 그러나 이러한 건강기능 작용들에 대한 과학적인 근거가 충분히 구축되어 있지는 않아, 유럽식품 위생위원회(Euroupe Food Safety Authority)는 항생제 투여에 따른 설사증상의 완화와 같은 건강기능 표시를 금지하고 있어, 프로바이오틱스의 작용기전에 대한 연구가 필요한 시점이라 하겠다.

• Journal of Microbiology
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• 제43권4호
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• pp.345-353
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• 2005

The complex ecosystem of intestinal micro flora is estimated to harbor approximately 400 different microbial species, mostly bacteria. However, studies on bacterial colonization have mostly been based on culturing methods, which only detect a small fraction of the whole microbiotic ecosystem of the gut. To clarify the initial acquisition and subsequent colonization of bacteria in an infant within the few days after birth, phylogenetic analysis was performed using 16S rDNA sequences from the DNA iso-lated from feces on the 1st, 3rd, and 6th day. 16S rDNA libraries were constructed with the amplicons of PCR conditions at 30 cycles and $50^{\circ}C$ annealing temperature. Nine independent libraries were produced by the application of three sets of primers (set A, set B, and set C) combined with three fecal samples for day 1, day 3, and day 6 of life. Approximately 220 clones ($76.7\%$) of all 325 isolated clones were characterized as known species, while other 105 clones ($32.3\%$) were characterized as unknown species. The library clone with set A universal primers amplifying 350 bp displayed increased diversity by days. Thus, set A primers were better suited for this type of molecular ecological analysis. On the first day of the life of the infant, Enterobacter, Lactococcus lactis, Leuconostoc citreum, and Streptococcus mitis were present. The largest taxonomic group was L. lactis. On the third day of the life of the infant, Enterobacter, Enterococcus faecalis, Escherichia coli, S. mitis, and Streptococcus salivarius were present. On the sixth day of the life of the infant, Citrobacter, Clostridium difficile, Enterobacter sp., Enterobacter cloacae, and E. coli were present. The largest taxonomic group was E. coli. These results showed that microbiotic diversity changes very rapidly in the few days after birth, and the acquisition of unculturable bacteria expanded rapidly after the third day.

• 한국식품위생안전성학회지
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• 제29권2호
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• pp.85-91
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• 2014

Deoxynivalenol (DON) and related trichothecene mycotoxins are extensively distributed in the cereal-based food and feed stuffs worldwide. Recent climate changes and global grain trade increased chance of exposure to more DON and related toxic metabolites in poorly managed production systems. Monitoring the biological and environmental exposures to the toxins are crucial in protecting human and animals from toxicities of the hazardous contaminants in food or feeds. Exposure biomarkers including urine DON itself are prone to shift to less harmful metabolites by intestinal microbiota and liver metabolic enzymes. De-epoxyfication of DON by gut microbes such as Eubacterium strain BBSH 797 and Eubacterium sp. DSM 11798 leads to more fecal secretion of DOM-1. By contrast, most of plant-derived DON-glucoside is also easily catabolized to free DON by gut microbes, which produces more burden to body. Phase 2 hepatic metabolism also contributes to the glucuronidation of DON, which can be useful urine biomarkers. However, chemical modification could be very typical depending on the anthropologic or genetic background, luminal bacteria, and hepatic metabolic enzyme susceptibility to the toxins in the diet. After toxin exposure, effect biomarkers are also important in estimating the linkage and mechanisms of foodborne diseases in human and animal population. Most prominent adverse effects are demonstrated in the DON-induced immunological and behavioral disorders. For instance, acutely elevated interleukin-8 from insulted gut exposed to dietaty DON is a dominant clinical biomarker in human and animals. Moreover, subchronic exposure to the toxins is associated with high levels of serum IgA, a biological mediator of IgA nephritis. In particular, anorexia monitoring using mouse models are recently developed to monitor the biological activities of DON-induced feed refusal. It is also mechanistically linked to alteration of serotoin and peptide YY, which are promising biomarkers of neurological disorders by the toxins. As animal-alternative biomonitoring, huamn enterocyte-based assay has been developed and more realistic gut mimetic models would be useful in monitoring the effect biomarkers in resposne to toxic contaminants in the future investigations.

• 한국식품과학회지
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• 제53권6호
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• pp.775-784
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• 2021

Clostidium butyricum을 분리하기 위하여 국내 모유수유 신생아 분변으로부터 혐기성 균주를 선별하였고 버블을 생성하는 100개의 균을 확보하였다. 이중 Clostridium perfringens에 대한 항균력과 butyric acid의 생산이 가장 우수한 DIMO 52 균주를 선발하였고, 형태학적 특성, 생리 생화적 특성 및 16S rRNA 유전자 분석을 통하여 C. butyricum으로 동정되어 C. butyricum DIMO 52로 명명하였다. 성장률, butyric acid 생산 및 pH 변화를 배양 36시간 동안 모니터링하였다. 배양 24시간 후 DIMO 52 균주의 최대 성장에 도달하였고, butyric acid 최대 농도는 대략 34.73±4.27 mM이었으며, pH는 7.2에서 2.5로 변경되었다. DIMO 52 균주는 낮은 pH와 oxgall에 높은 저항성이 있다. pH 2에서 2시간 동안 접종의 약 67.5%의 유의성 있는 생존율을 보였다(p<0.05). 그리고, 0.3% oxgall이 함유된 RCM 액체배지에서 24시간 동안 접종의 약 64.9%의 유의성 있는 생존율을 보였다(p<0.05). 또한, DIMO 52은 Escherichia coli KCTC 2441와 Salmonella Typhimurium KCTC 1925에 대해 억제효과를 나타냈다. 두 균주에 대한 항균효과는 아마도 butyric acid에 의한 낮은 pH 때문인 것으로 보였다. 5×10³ CFU/mL 생균수 까지는 RAW264.7 세포에 세포독성이 없는 것으로 관찰되었고, NO 생성을 억제할 수 있는 최저 균수를 확인한 결과 약 1×10³ CFU/mL 생균수에서 LPS만 처리한 군 대비 약 33%의 NO 생성을 억제하는 것으로 분석되었다(p<0.01). 이 결과는 C. butyricum DIMO 52이 NO radical 소거 및 항염증 활성을 가지고 있음을 시사한다. 결론적으로, 본 연구에서 분리된 C. butyricum DIMO 52의 프로바이오틱스 특성을 확인하였다.