• 제목/요약/키워드: Co-incubated

검색결과 354건 처리시간 0.032초

AcuD Gene Knockout Attenuates the Virulence of Talaromyces marneffei in a Zebrafish Model

  • Feng, Jiao;Chen, Zhiwen;He, Liya;Xiao, Xing;Chen, Chunmei;Chu, Jieming;Mylonakis, Eleftherios;Xi, Liyan
    • Mycobiology
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    • 제47권2호
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    • pp.207-216
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    • 2019
  • Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at $28^{\circ}C$, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ${\Delta}AcuD$ was significantly attenuated in this Zebrafish infection model.

Effect of bicarbonate and progesterone on plasma membrane integrity, acrosome reaction and proportion of fatty acids in boar sperm

  • Park, Choon-Keun;Lee, Sang-Hee
    • 한국동물생명공학회지
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    • 제37권3호
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    • pp.202-208
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    • 2022
  • This study investigated the influence of sodium bicarbonate (NaHCO3) and progesterone on acrosome reaction and proportion of polyunsaturated fatty acid (PUFA) composition boar sperm. The sperm were diluted with semen extender and incubated with NaHCO3 and progesterone at 38℃, 5% CO2 for 6 h. Plasma membrane integrity and acrosome reaction were analyzed using SYBR14/propidium iodide (PI) and FITC-PNA/PI doubling staining method, and proportion of PUFA was analyzed using gas chromatography. In results, Plasma membrane integrity was significantly decreased in 50 mM NaHCO3 group and acrosome reaction was significantly increased by over the 100 mM NaHCO3 group compared to control group (p < 0.05). In addition, progesterone significantly increased decreased plasma membrane integrity at 100 mM progesterone and acrosome reaction at over the 5.0 µM progesterone (p < 0.05), but there was no difference among the 5.0 to 100 µM groups. PUFAs were significantly decreased in 100 mM NaHCO3 and 50 µM progesterone treatments compared to control group. In summary NaHCO3 and progesterone induce acrosome reaction and reduce PUFA composition in boar sperm, therefore, the results maybe help to understand basically knowledge for the acrosome reaction and PUFA composition in boar sperm.

Effect of Bacteriocin-Like Inhibitory Substance (BLIS) from Enterococcus faecium DB1 on Cariogenic Streptococcus mutans Biofilm Formation

  • Kim, Ni-Na;Kim, Bong Sun;Lee, Han Bin;An, Sunghyun;Kim, Donghan;Kang, Seok-Seong
    • 한국축산식품학회지
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    • 제42권6호
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    • pp.1020-1030
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    • 2022
  • The aim of the study was to investigate the effect of bacteriocin-like inhibitory substance (BLIS) from Enterococcus faecium DB1 on cariogenic Streptococcus mutans biofilm. Crystal violet staining, fluorescence, and scanning electron microscopy analyses demonstrated that the BLIS from Enterococcus faecium DB1 (DB1 BLIS) inhibited S. mutans biofilm. When DB1 BLIS was co-incubated with S. mutans, biofilm formation by S. mutans was significantly reduced (p<0.05). DB1 BLIS also destroyed the preformed biofilm of S. mutans. In addition, DB1 BLIS decreased the viability of S. mutans biofilm cells during the development of biofilm formation and in the preformed biofilm. DB1 BLIS significantly decreased the growth of S. mutans planktonic cells. Furthermore, S. mutans biofilm on the surface of saliva-coated hydroxyapatite discs was reduced by DB1 BLIS. Taken together, DB1 BLIS might be useful as a preventive and therapeutic agent against dental caries caused by S. mutans.

Involvement of NOX2-derived ROS in human hepatoma HepG2 cell death induced by Entamoeba histolytica

  • Young Ah Lee ;Myeong Heon Shin
    • Parasites, Hosts and Diseases
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    • 제61권4호
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    • pp.388-396
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    • 2023
  • Entamoeba histolytica is an enteric tissue-invasive protozoan parasite causing amoebic colitis and liver abscesses in humans. Amoebic contact with host cells activates intracellular signaling pathways that lead to host cell death via generation of caspase-3, calpain, Ca2+ elevation, and reactive oxygen species (ROS). We previously reported that various NADPH oxidases (NOXs) are responsible for ROS-dependent death of various host cells induced by amoeba. In the present study, we investigated the specific NOX isoform involved in ROS-dependent death of hepatocytes induced by amoebas. Co-incubation of hepatoma HepG2 cells with live amoebic trophozoites resulted in remarkably increased DNA fragmentation compared to cells incubated with medium alone. HepG2 cells that adhered to amoebic trophozoites showed strong dichlorodihydrofluorescein diacetate (DCF-DA) fluorescence, suggesting intracellular ROS accumulation within host cells stimulated by amoebic trophozoites. Pretreatment of HepG2 cells with the general NOX inhibitor DPI or NOX2-specific inhibitor GSK 2795039 reduced Entamoeba-induced ROS generation. Similarly, Entamoeba-induced LDH release from HepG2 cells was effectively inhibited by pretreatment with DPI or GSK 2795039. In NOX2-silenced HepG2 cells, Entamoeba-induced LDH release was also significantly inhibited compared with controls. Taken together, the results support an important role of NOX2-derived ROS in hepatocyte death induced by E. histolytica.

체외성숙 돼지난포란의 체외수정과 배발달에 관한 연구 II. 각종 배양액, 돼지난구세포 및 생쥐태아간세포와의 공동배양이 체외수정 돼지 난포란의 체외발달에 미치는 영향 (Studies on In Vitro Fertilization and Development of In Vitro Matured Porcine Follicular Oocytes I. Effect of Various Media and Co-culture with Porcine Cumulus Cellsor Mouse Fetal Fibroblast Cells on In Vitro Development of In Vitro Fertilized Oocytes)

  • 정형민;엄상준;승경록;이상준;이훈택;정길생
    • 한국가축번식학회지
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    • 제17권2호
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    • pp.113-120
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    • 1993
  • 본 연구는 제외생산된 돼지 수정란의 처1외발생율을 제고하기 위하여 각종 배양액파 돼지난구세포 혹은 생 쥐태아간세포와의 공동배양 효과플 조사하였다 m-KRB, BECM 및 TCM-HEPES 배양액을 공시하 여 제외수정란을 배양한 결과 배반포기까지 발달하는 비율은 전처리구에서 0~1.0%로써 극히 저조하였다. 특히 대부분의 수정란은 4-세포기 단계에서 발달이 정지되었다. 한편, 단층세포가 유도된 돼지 난구세포나 생쥐 태아간세포와 함께 제외수정란을 공동배양한 결파 2, 4-, 8~16-, 32-세포기, 상실배가 빛 배반포로 받달하는 비율은 각각 61.1~67.0%, 59.0~58.0%, 42.5~43.1%, 28.4~30.2% 및 20.4~21.0%였다 이러한 결파는 단순배양액에서 체외배양한 수정란의 발탄 성적 보다유의하게 높은 것이었다. 이상의 결과를 종합하여 볼 때 1세포기 수정란을 체외에서 배양할때 체세포와의 공동배양은 수정란의 체외발달을 촉진하는 것으로 생각된다.

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살균제 $^{14}C$-propiconazole의 토심별 분해 (Degradation of $^{14}C$-propiconazole in soil from different depths)

  • 안득현;김인선;서용택
    • 농약과학회지
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    • 제3권2호
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    • pp.8-18
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    • 1999
  • 살균제 $^{14}C$-propiconazole의 토심별 분해를 멸균 및 비멸균 조건의 토양에서 조사하였다. Propiconazole을 토양에 7.55 mg/kg의 수준으로 처리한 후 배양기간 동안 발생된 $^{14}CO_{}2$ 양, 유기용매 추출 및 추출불가 잔류량, 그리고 분해산물을 등을 조사하였다. 배양기간 20주 동안 방출된 약제의 $^{14}CO_{2}$는 멸균 토양의 경우 토심에 따라 $0.7{\sim}1.3%$ 수준이었으며 비멸균 토양의 경우 $4.8{\sim}7.6%$이었다. 토양중 유기용매 추출불가 방사능은 멸균 토양의 경우 토심에 따라 $11.2{\sim}22.12%$ 수준이었으며 비멸균 토양의 경우 $22.1{\sim}41.9%$로서 비멸균 토양에서 더 높았다. 토양중 추출불가 잔류량은 주로 humin에 분포하였으며 배양시간에 따라 증가하였다. 토양중 propiconazole의 주요 분해산물은 수산화 치환된 형태 및 ketone성 화합물이었으며 멸균 토양에서는 모화합물만이 검출되었다. Propiconazole의 토양중 휘발성 물질 및 $CO_{2}$ 생성량과 잔류 및 분해특성은 이 화합물이 화학적 및 생물학적 분해에 안정함을 시사하였다.

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항원성이 저감화 처리된 국내산 밀을 이용한 발효빵의 제조 및 품질 특성 (Development and Quality Evaluation of Hypo-Allergenic Bakery Products using Homegrown Wheat)

  • 박주연;안정엽;한영숙
    • 한국식품과학회지
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    • 제37권5호
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    • pp.736-740
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    • 2005
  • Yeast로 발효시킨 빵을 섭취한 후 호흡기 증상이 나타났다는 보고에서 yeast가 생성한 ${\alpha}-amylase$가 알레르기의 원인(22,23)임이 밝혀졌으므로 본 연구에서는 yeast 대체 첨가제로 sodium bicarbonate를 선택하였으며, 첨가제의 농도가 다른 12종류의 반죽을 제조하였다. Meissle 발효관에 의한 $CO_2$ 발생량 비교 실험 결과, sodium bicarbonate 첨가를 많이 할수록 $CO_2$ 발생량이 증가함을 알 수 있었다. 그러나 hypo-allergenic breads(HAB) 제조 시 2.0% 이상 첨가한 HAB 7 이상에서는 sodium bicarbonate의 쓴맛이 강하게 나타나 관능 특성 평가 결과 기호도가 떨어지는 것으로 드러났다. 체적과 무게에 대한 비로 빵의 품질을 조사한 결과, 체적의 경우 hypo-allergenic bread (HAB) 11, 12가 가장 높은 값을 보였다. HAB 12, 9, 11이 가장 많이 부푼 빵이 되었으며 HAB 1은 빵의 부피가 현저하게 적어 제빵용 조합으로는 부적합하였다. 관능검사에서 색, 향, 맛 항목에서 유의적인 차이를 보였다. 색은 HAB 7이, 향과 맛 에서는 HAB 5가 가장 우수한 결과를 나타내었으며 기공과 촉감항목은 유의적인 차이를 나타내지 않았다. 마지막으로 전체적인 기호도 평가 시 매우 유의적인 차이를 보이며 HAB 5가 우수하게 나타났다. 결과적으로 hypo-allergenic bread(HAB) 시작품은 b.p. protease로 gluten을 분해하여 항원성을 저감화 시킨 국내산 밀 반죽에 sodium bicarbonate 1.5%, sucrose 1.0%, NaCl 1.0%, citric acid 1.0%를 첨가하여 제작되었다.

백굴채(白屈菜)가 천식유발 생쥐의 폐조직에서 호산구 증식과 면역 세포 활성화에 미치는 영향 (The Effects of Chelidonii Herba on the Proliferation of Eosinophils and Activation of Immuno-cells in Asthma-induced Mouse)

  • 노성수
    • 대한본초학회지
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    • 제23권2호
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    • pp.99-109
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    • 2008
  • Objectives : The present study was to investigate the effect of extract of Chelidonii herba (ECH) on the proliferation and activation of eosinophils which were prepared from lung cells of asthma-induced mouse by ovalbumin (OVA) treatment. Methods : C57BL/6 mouse was exposed to OVA three times a week for 6 weeks. The mouse lung tissues were dissected out, chopped and dossiciated with collagenase (1 ${\mu}g$/ml). Eosinophils were activated by rIL-3/rmIL-5 co-treatments. The lung cells were treated with ECH, incubated for 48 hr at $37^{\circ}C$, and analyzed by flow cytometery, ELISA, RT-PCR, and immuno-histochemical analysis. Results : In FACS analysis, number of granulocyte/lymphocyte, $CD3e^-$/$CCR3^+$, $CD3e^+$/$CD69^+$, $CD4^+$ and $CD23^+$/$B220^+$ in asthma-induced lung cells were significantly decreased by ECH treatment compared to the control group. And mRNA expression for IL-4, IL-5, IL-13, CCR3 and eotaxin in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by ECH treatment. In ELISA analysis, production levels of IL-3, IL-5, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by ECH treatment. ECH treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-ECH treated control cells. Immunohistochemical analysis revealed that ECH treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusions : The present data suggested that Chelidonium majus L. may have an effect on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Chelidonium majus L.

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Standardized rice bran extract improves hepatic steatosis in HepG2 cells and ovariectomized rats

  • Lim, Dong Wook;Jeon, Hyejin;Kim, Minji;Yoon, Minseok;Jung, Jonghoon;Kwon, Sangoh;Cho, Suengmok;Um, Min Young
    • Nutrition Research and Practice
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    • 제14권6호
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    • pp.568-579
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    • 2020
  • BACKGROUD/OBJECTIVES: Hepatic steatosis is the most common liver disorder, particularly in postmenopausal women. This study investigated the protective effects of standardized rice bran extract (RBS) on ovariectomized (OVX)-induced hepatic steatosis in rats. MATERIALS/METHODS: HepG2 cells were incubated with 200 µM oleic acid to induce lipid accumulation with or without RBS and γ-oryzanol. OVX rats were separated into three groups and fed a normal diet (ND) or the ND containing 17β-estradiol (E2; 10 ㎍/kg) and RBS (500 mg/kg) for 16 weeks. RESULTS: RBS supplementation improved serum triglyceride and free fatty acid levels in OVX rats. Histological analysis showed that RBS significantly attenuated hepatic fat accumulation and decreased hepatic lipid, total cholesterol, and triglyceride levels. Additionally, RBS suppressed the estrogen deficiency-induced upregulation of lipogenic genes, such as sterol regulatory element-binding protein 1 (SREBP1), acetyl-CoA carboxylase 1, fatty acid synthase, glycerol-3-phosphate acyltransferase, and stearoyl-CoA desaturase 1. CONCLUSIONS: RBS and γ-oryzanol effectively reduced lipid accumulation in a HepG2 cell hepatic steatosis model. RBS improves OVX-induced hepatic steatosis by regulating the SREBP1-mediated activation of lipogenic genes, suggesting the benefits of RBS in preventing fatty liver in postmenopausal women.

An Ototoxic Antibiotic Gentamicin Can Increase PKA-caveolin-1 Signaling Pathway in Differentiated Vestibular Cell Line (UB/UE-1)

  • Kim, Kyu-Sung;Cho, Byung-Han;Choi, Ho-Seok;Park, Chang-Shin;Jung, Yoon-Gun;Kim, Young-Mo;Jang, Tae-Young
    • Molecular & Cellular Toxicology
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    • 제4권3호
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    • pp.177-182
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    • 2008
  • Caveolin proteins are mediators of cell death or the survival of injured cells, and they are inhibitors of various signaling pathways. The expression of caveolin-, which is involved in the protein kinase A (PKA) signaling pathway, was examined in the differentiated mouse vestibular cell line UB/UE-1 after gentamicin ototoxicity. Caveolae in the vestibular hair cell of healthy guinea pigs were observed through an electron microscope. UB/UE-1 cells were cultured at 95% $CO_2$ with 5% $O_2$ at $33^{\circ}C$ for 48 hours and at 95% $CO_2$ with 5% $O_2$ at $39^{\circ}C$ for 24 hours for differentiation. Cells were treated with 1 mM gentamicin, 0.02 mM H89 (PKA inhibitor), and then incubated for 24 hours. Caveolin-1 expression was examined by western blotting and PKA activity by a $PepTag^{(R)}$ assay. Caveolae were observed in the vestibular hair cells of healthy guinea pigs by electron microscopy. Caveolin-1 was expressed spontaneously in differentiated UB/UE-1 cells and increased after gentamicin treatment. PKA was also over-activated by gentamicin treatment. Both gentamicin-induced caveolin-1 expression and PKA over-activation were inhibited by H89. These results indicate that gentamicin-induced caveolin-1 expression is mediated by the PKA signaling pathway. We conclude that caveolae/ caveolin activity, induced via a PKA signaling pathway, may be one of the mechanisms of gentamicin-induced ototoxicity.