• Title/Summary/Keyword: GC-1 cell

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Metabolome-Wide Reprogramming Modulated by Wnt/β-Catenin Signaling Pathway

  • Soo Jin Park;Joo-Hyun Kim;Sangtaek Oh;Do Yup Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.114-122
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    • 2023
  • A family of signal transduction pathways known as wingless type (Wnt) signaling pathways is essential to developmental processes like cell division and proliferation. Mutation in Wnt signaling results in a variety of diseases, including cancers of the breast, colon, and skin, metabolic disease, and neurodegenerative disease; thus, the Wnt signaling pathways have been attractive targets for disease treatment. However, the complicatedness and large involveness of the pathway often hampers pinpointing the specific targets of the metabolic process. In our current study, we investigated the differential metabolic regulation by the overexpression of the Wnt signaling pathway in a timely-resolved manner by applying high-throughput and un-targeted metabolite profiling. We have detected and annotated 321 metabolite peaks from a total of 36 human embryonic kidney (HEK) 293 cells using GC-TOF MS and LC-Orbitrap MS. The un-targeted metabolomic analysis identified the radical reprogramming of a range of central carbon/nitrogen metabolism pathways, including glycolysis, TCA cycle, and glutaminolysis, and fatty acid pathways. The investigation, combined with targeted mRNA profiles, elucidated an explicit understanding of activated fatty acid metabolism (β-oxidation and biosynthesis). The findings proposed detailed mechanistic biochemical dynamics in response to Wnt-driven metabolic changes, which may help design precise therapeutic targets for Wnt-related diseases.

Determination of Gasoline Brands by the Comparison of Infrared Spectra of Polymeric Dispersants (청정분산제의 적외선 분광스펙트럼 비교를 통한 자동차용 휘발유 제조사의 판별)

  • Kim, Myeonghee;Jang, Youngsik;Jung, Chungseop;Lee, Hyunkee
    • Analytical Science and Technology
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    • v.11 no.6
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    • pp.469-473
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    • 1998
  • The gasoline brands can be determined by gas chromatography. However, determining from the differences in chromatograms is sometimes ambiguous because the gasoline composition is becoming similar from refinery to refinery due to stringent regulations for the protection of the atmospheric environment. To determine the gasoline brands of five refineries in Korea, we have obtained and compared IR spectra of polymeric dispersants which are added to gasoline at several hundreds of ppm levels. Since the deposit control additives used by the five refineries in Korea are different from one another, it is possible to determine the gasoline brands by comparing their IR spectra. A strong and broad C-O stretch absorption peak appears at $1,096cm^{-1}$ for the additives used by A, B, and C refineries, which renders an easy differentiation of the additives from those of D and E refineries. The differentiation of all five gasoline brands are possible due to the characteristic vibrations present in each additive.

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Foeniculum vulgare essential oil nanoemulsion inhibits Fusarium oxysporum causing Panax notoginseng root-rot disease

  • Hongyan Nie;Hongxin Liao;Jinrui Wen;Cuiqiong Ling;Liyan Zhang;Furong Xu;Xian Dong
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.236-244
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    • 2024
  • Background: Fusarium oxysporum (F. oxysporum) is the primary pathogenic fungus that causes Panax notoginseng (P. notoginseng) root rot disease. To control the disease, safe and efficient antifungal pesticides must currently be developed. Methods: In this study, we prepared and characterized a nanoemulsion of Foeniculum vulgare essential oil (Ne-FvEO) using ultrasonic technology and evaluated its stability. Traditional Foeniculum vulgare essential oil (T-FvEO) was prepared simultaneously with 1/1000 Tween-80 and 20/1000 dimethyl sulfoxide (DMSO). The effects and inhibitory mechanism of Ne-FvEO and T-FvEO in F. oxysporum were investigated through combined transcriptome and metabolome analyses. Results: Results showed that the minimum inhibitory concentration (MIC) of Ne-FvEO decreased from 3.65 mg/mL to 0.35 mg/mL, and its bioavailability increased by 10-fold. The results of gas chromatography/mass spectrometry (GC/MS) showed that T-FvEO did not contain a high content of estragole compared to Foeniculum vulgare essential oil (FvEO) and Ne-FvEO. Combined metabolome and transcriptome analysis showed that both emulsions inhibited the growth and development of F. oxysporum through the synthesis of the cell wall and cell membrane, energy metabolism, and genetic information of F. oxysporum mycelium. Ne-FvEO also inhibited the expression of 2-oxoglutarate dehydrogenase and isocitrate dehydrogenase and reduced the content of 2-oxoglutarate, which inhibited the germination of spores. Conclusion: Our findings suggest that Ne-FvEO effectively inhibited the growth of F. oxysporum in P. notoginseng in vivo. The findings contribute to our comprehension of the antifungal mechanism of essential oils (EOs) and lay the groundwork for the creation of plant-derived antifungal medicines.

Cryoprotective Properties of Exopolysaccharide (P-21653) Produced by the Antarctic Bacterium, Pseudoalteromonas arctica KOPRI 21653

  • Kim, Sung-Jin;Yim, Joung-Han
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.510-514
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    • 2007
  • Twenty-five bacterial strains that secrete mucous materials were isolated from sediment obtained from King George Island, Antarctica. Seven of these strains proved capable of producing cryoprotective exopolysaccharides. The strain KOPRI 21653 was selected for the further study of an anti-ice-nucleating polysaccharide (ANP), which originated from a polar region. KOPRI 21653 was identified as Pseudoalteromonas arctica as the result of 16S rRNA analysis. The exopolysaccharide, P-21653, was purified completely from the KOPRI 21653 cell culture via column chromatography and protease treatment. The principal sugar components of P-21653 were determined to be galactose and glucose, at a ratio of 1:1.5, via GC-MS analysis. The cryoprotective activity of P-21653 was characterized via an E. coli viability test. In the presence of 0.1% (w/v) P-21653, the survival ratio of E. coli cells was as high as 82.6% over three repeated freeze-thaw cycles. The survival ratio decreased drastically to 71.5 and 48.1 %, respectively, in five and seven repeated cycle conditions; however, the survival ratios were greater over three (96.6-92.1%) to seven (100.5-91.6%) freeze-thaw cycles in the presence of 0.5 and 1.0% (w/v) P-21653. In addition, at much lower concentrations (0.1-1.0%), P-21653 resulted in survival ratios (83.1-98.4%) similar to those of two commercially available cryoprotectants ($V_{EG}$ plus X-1000, 92.9% and $V_{M3}$, 95.3%), which were utilized at the recommended concentrations (90%). The biochemical characteristics of exopolysaccharide P-21653 reflect that this compound may be developed as a useful cryoprotectant for use in medical applications and in the food industry.

Isolation and Characterization of a Crude oil-Degrading Strain, Nocardia sp. H 17-1 (원유 분해균주 Nocardis sp. Hl7-1의 분리 및 특성)

  • 이창호;권기석
    • KSBB Journal
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    • v.11 no.6
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    • pp.654-662
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    • 1996
  • Bacterial strains which degrade crude oil were isolated by liquid culture from oil-spilled soil, and four isolates were selected among them. The strain Hl7-1 was finally selected after testing emulsifying activity and oil conversion rate. The strain Hl7-1 was identified as a Nocardia sp. based on the test for morphological, biochemical and physiological characteristics. It appears to be highly specialized for growth on crude oil in minimal salts medium since it showed preference for oil or degradation products as substrates for growth. It was found that it could grow on at least fifteen different hydrocarbons. The optimum cultural and environmental conditions were seeked. Cell growth and emulsification activity as a function of time were also determined. Crude oil degradation and the reduction of product peak was identified by the analysis of remnant oil by gas chromatography after 3 days of cultivation. Approximately 83% of oil were converted into a form no longer extractable by organic solvents.

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Effect of the Bifunctional Chelate on the Biodistribution of 99mTc-labeled Cyclic RGD Peptide

  • Lee, Dong-Eun;Choi, Kang-Hyuk
    • Journal of Radiation Industry
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    • v.12 no.4
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    • pp.355-363
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    • 2018
  • A novel $N_3S_1$ chelate, Pro-Lys-Cys (PKC) to cyclic RGD to radiolabel with $^{99m}Tc$ was conjugated in an effort to decrease the high intestinal accumulation observed for $^{99m}Tc$-labeled PGC-RGD. The target specificity of the resulting PKC-RGD was similar to that of PGC-RGD as determined by a cell binding assay and a competition binding assay. The $^{99m}Tc$ radiolabeling of PKC-RGD resulted in radiochemical yields of 98% under mild conditions at high specific activities. Biodistribution data in normal mice clearly showed a significant decrease in intestinal uptake at 2 h postinjection for the $^{99m}Tc-PKC-c$ (RGDyK) compared to the $^{99m}Tc-GC-c$ (RGDyK) (from $19.65%ID{\cdot}g^{-1}$ to $7.31%ID{\cdot}g^{-1}$ for the GI tract). The $^{99m}Tc-PKC-c$ (RGDyK) biodistribution was also shown by a higher retention of radioactivity in the whole body, but with kidney accumulation over 8-fold higher than observed with $^{99m}Tc-PGC-c$ (RGDyK) at 2 h ($12.62%ID{\cdot}g^{-1}$ for PKC-RGD and $1.54%ID{\cdot}g^{-1}$ for PGC-RGD, respectively). These results show that the biodistribution may be altered especially concerning lipophilicity resulting in renal rather than hepatobiliary excretion. This comparative study made it possible to explore the effects of lipophilicity on the biodistribution of $^{99m}Tc$-labeled c (RGDyK) through the use of different tripeptide $N_3S_1$ chelators. Therefore, $^{99m}Tc-PKC-c$ (RGDyK) may be an attractive alternative for the in vivo imaging of integrin receptors.

Effects of Ganoderma lucidum Extract on Production of Fatty Acids by Rhodotorula glutinis (영지 열수추출물이 Rhodotorula glutinis의 지방산 생성에 미치는 영향)

  • Joo, Hyun-Kyu;Park, Woo-Chul;Sa, Tong-Min;Lee, Young-Tack;Yun, Choong-Hyo
    • Korean Journal of Food Science and Technology
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    • v.29 no.1
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    • pp.120-125
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    • 1997
  • Rhodotorula glutinis was inoculated in the medium containing 0 ($S_0$), 0.01 ($S_1$), 0.1 ($S_2$) and 1.0% ($S_3$) Ganodoma lucidum extract and incubated in a shaking incubator at $30^{\circ}C$ for 8 days and the cell growth, sugarity, lipid content, and fatty acid composition were measured to investigate the effects of G. lucidum extract on the growth and biosynthesis of fatty acids by oleaginous yeast, Rhodotorula glutinis. After the 8 day incubation, the cell growth of $S_1,\;S_2\;and\;S_3$ increased 1.6, 1.7 and 2.1 times, respectively, than that of $S_0$. Sugar consumption of incubating medium was decreased but the crude lipid content in R. glutinis was increased with increase of the amount of G. Lucidum extract. Myristic, palmitic, stearic, oleic and linoleic acids were identified by GC as the major fatty acids in the crude lipid produced by R. glutinis and the content of unsaturated fatty acids (38.6 mg/g) was greater than that of saturated fatty acids (22.3 mg/g). As the G. lucidum extract concentration increased, fatty acids contents were increased except myristic acid, and the most increase occurred at the addition of 0.1% while they were considerably decreased in the case of the addition of 1.0% G. lucidum extract.

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Propagation of Human Embryonic Stem Cells on Human Amniotic Fluid Cells as Feeder Cells in Xeno-Free Culture Conditions

  • Jung, Juwon;Baek, Jin Ah;Seol, Hye Won;Choi, Young Min
    • Development and Reproduction
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    • v.20 no.1
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    • pp.63-71
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    • 2016
  • Human embryonic stem cells (hESCs) have been routinely cultured on mouse embryonic fibroblast feeder layers with a medium containing animal materials. For clinical application of hESCs, animal-derived products from the animal feeder cells, animal substrates such as gelatin or Matrigel and animal serum are strictly to be eliminated in the culture system. In this study, we performed that SNUhES32 and H1 were cultured on human amniotic fluid cells (hAFCs) with KO-SR XenoFree and a humanized substrate. All of hESCs were relatively well propagated on hAFCs feeders with xeno-free conditions and they expressed pluripotent stem cell markers, alkaline phosphatase, SSEA-4, TRA1-60, TRA1-81, Oct-4, and Nanog like hESCs cultured on STO or human foreskin fibroblast feeders. In addition, we observed the expression of nonhuman N-glycolylneuraminic acid (Neu5GC) molecules by flow cytometry, which was xenotransplantation components of contamination in hESCs cultured on animal feeder conditions, was not detected in this xeno-free condition. In conclusion, SNUhES32 and H1 could be maintained on hAFCs for humanized culture conditions, therefore, we suggested that new xeno-free conditions for clinical grade hESCs culture will be useful data in future clinical studies.

Change of Steroid Receptor Number of and Bioactivity of Gonadotropins in the Follicular Fluid of Porcine Ovarian Atretic Follicles(II):Testosterone Receptor (돼지 폐쇄여포내 스테로이드 수용수체의 변화와 여포액내 생식소 자극호르몬의 활성도 변화(II): 테스토스테론 수용체)

  • Yoon, Yong-Dal;Lee, Chang-Joo;Chun, Eun-Hyun;Kim, Moon-Kyoo;Lee, Joon-Yeong
    • Clinical and Experimental Reproductive Medicine
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    • v.16 no.1
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    • pp.23-34
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    • 1989
  • Estradiol (E)은 난소내 과립세포 (granulosa cell, GC)를 증가시키고 생식소 자극호르몬과 협동으로 배란을 유도한다. Androgen은 E의 작용과 반대의 작용을 나타내며 여포의 폐쇄요인으로 알려지고 있다. Testosterone(T)이 폐쇄여포의 여포액내 다량 존재하는 것이 알려짐에 따라 난소내에도 자가조절분비(autocrine)또는 paracrine regulation에 의해 작용을 나타낼 것으로 가정되어 난소내 여포가 폐쇄됨에 따라 그 수용체의 변화하는 양상을 조사하고져 하였다. 흰쥐의 과립세포의 세포질에는 $51.3{\pm}6.1$fmol/mg protein의 Estradiol 수용체(ER) ; $153.1{\pm}25.3$의 Testosterone수용체(TR) ; 또한 $35.1{\pm}8.1$의 Progesterone수용체(PR)가 존재하였다. 과립세포내 ER용 세포질내 E를 제거한 후에 정량이 가능하였고 또한 과립세포내에도 TR이 사람에서는 $23.4{\pm}7.2$ fmol/mg protein, 돼지는 $98.5{\pm}23.1$로 상당량 존재함을 관찰하였다. Dihydrotestosterone Enanthate(DHTE)를 100ug/흰쥐의 농도로 처리한 결과 난소내 TR의 농도는 변화가 없이 ER의 농도만 현저히 저하되고 쥐의 난소무게 역시 감소하는 것을 발견하였다. 위의 결과로 보아 난소내에도 스테로이드 호르몬은 autocrine(자가조절)방법으로 작용하며 An-drogen이 난소의 무게를 감소시키는 것은 ER의 수를 감소시켜 E의 작용이 억제되고 여포들이 폐쇄를 일으켜 그 증식이 저하된 때문으로 사료된다.

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Evaluation on Anti-fungal Activity and Synergy Effects of Essential Oil and Their Constituents from Abies holophylla (전나무 정유의 항진균 효과와 유효성분의 시너지효과 평가)

  • Kim, Seon-Hong;Lee, Su-Yeon;Cho, Seong-Min;Hong, Chang-Young;Park, Mi-Jin;Choi, In-Gyu
    • Journal of the Korean Wood Science and Technology
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    • v.44 no.1
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    • pp.113-123
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    • 2016
  • This study was to investigate the antifungal activity of A. holophylla essential oil against dermatophytes, such as Epidermophyton floccosum, Trichophyton mentagrophytes and Trichophyton rubrum, and to determine the potential effective compound as dermatitis treatment. To evaluate the potential antifungal activities of A. holophylla essential oil and its fractions, paper disc diffusion and agar dilution method tested with morphological observation. Also, their major constituents were analyzed by GC/MS. To determine synergic effects of active ingredient from A. holophylla essential oil were carried out by checkerboard microtiter plate testing. The morphological changes of the dermatophytes exposed to active fraction G4 were observed by electron microscopes. As the results, the highest activities were identified in the fraction containing ${\alpha}$-bisabolol. A mixture of ${\alpha}$-bisabolol and bornyl acetate showed the synergy effects, expressing high potential effects. Also, morphological observation using electron microscopes showed a dramatic changes of cell membrane of E. floccosum and T. rubrum exposed to fraction G4 containing ${\alpha}$-bisabolol. In conclusion, A. holophylla essential oil and its constituents were expected to be used as antifungal agent or raw material for dermatitis therapy.