• Title/Summary/Keyword: Mass Spectrometry (MS)

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Differentially Expressed Proteins in ER+ MCF7 and ER- MDA-MB-231 Human Breast Cancer Cells by RhoGDI-α Silencing and Overexpression

  • Hooshmand, Somayeh;Ghaderi, Abbas;Yusoff, Khatijah;Thilakavathy, Karuppiah;Rosli, Rozita;Mojtahedi, Zahra
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3311-3317
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    • 2014
  • Background: The consequence of Rho GDP dissociation inhibitor alpha (RhoGDI${\alpha}$) activity on migration and invasion of estrogen receptor positive ($ER^+$) and negative ($ER^-$) breast cancer cells has not been studied using the proteomic approach. Changes in expression of RhoGDI${\alpha}$ and other proteins interacting directly or indirectly with RhoGDI${\alpha}$ in MCF7 and MDA-MB-231, with different metastatic potentials is of particular interest. Materials and Methods: $ER^+$ MCF7 and ER- MDA-MB-231 cell lines were subjected to two-dimensional electrophoresis (2-DE) and spots of interest were identified by matrix-assisted laser desorption/ionization time of- flight/time-of-flight (MALDI-TOF/TOF) mass spectrometry (MS) analysis after downregulation of RhoGDI${\alpha}$ using short interfering RNA (siRNA) and upregulated using GFP-tagged ORF clone of RhoGDI${\alpha}$. Results: The results showed a total of 35 proteins that were either up- or down-regulated in these cells. Here we identifed 9 and 15 proteins differentially expressed with silencing of RhoGDI${\alpha}$ in MCF-7 and the MDA-MB-231 cells, respectively. In addition, 10 proteins were differentially expressed in the upregulation of RhoGDI${\alpha}$ in MCF7, while only one protein was identified in the upregulation of RhoGDI${\alpha}$ in MDA-MB-231. Based on the biological functions of these proteins, the results revealed that proteins involved in cell migration are more strongly altered with RhoGDI-${\alpha}$ activity. Although several of these proteins have been previously indicated in tumorigenesis and invasiveness of breast cancer cells, some ohave not been previously reported to be involved in breast cancer migration. Hence, these proteins may serve as useful candidate biomarkers for tumorigenesis and invasiveness of breast cancer cells. Conclusions: Future studies are needed to determine the mechanisms by which these proteins regulate cell migration. The combination of RhoGDI${\alpha}$ with other potential biomarkers may be a more promising approach in the inhibition of breast cancer cell migration.

Identification and Antifungal Susceptibility Profiles of Cyberlindnera fabianii in Korea

  • Park, Ji-Hyun;Oh, Junsang;Sang, Hyunkyu;Shrestha, Bhushan;Lee, Hyeyoung;Koo, Jehyun;Cho, Sung-Il;Choi, Ji Seon;Lee, Min-Ha;Kim, Jayoung;Sung, Gi-Ho
    • Mycobiology
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    • v.47 no.4
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    • pp.449-456
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    • 2019
  • Invasive fungal infections caused by Cyberlindnera fabianii have recently increased. However, biochemical kits such as API 20 C AUX and Vitek-2C have misidentified this species as other Candida spp. such as C. pelliculosa or C. utilis due to no information of Cy. fabianii in yeast database. During our 2016-2017 surveys, eleven isolates of Cy. fabianii were obtained in International St. Mary's Hospital in Korea. Here, we describe its morphological and molecular characteristics and tested its antifungal susceptibility against nine antifungal agents. The sequences of the ITS region and the D1/D2 region of LSU revealed 100% identity with the sequences of Cy. fabianii. In comparison with the results from MALDI-TOF mass spectrometry, we found that Cy. fabianii can be distinguished from other species. In antifungal susceptibility test, voriconazole and echinocandins exhibited good antifungal activities against the majority of Cy. fabianii isolates despite the absence of standard criteria.

Monitoring of Uniconazole by Using GC-ECD and GC-MSD (GC-ECD와 GC-MSD를 이용한 유니코나졸 잔류실태조사)

  • Kim, Eun-Jeung;Hong, Chae-Kyu;Choi, Su-Jeong;Lee, Yun-Jeong;Hwang, In-Sook;Kim, Moo-Sang;Chae, Young-Zoo
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.382-384
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    • 2013
  • We evaluated the presence of uniconazole residual pesticide in agricultural products by using multiclass pesticide multiresidue methods. Samples were collected from January to August, 2012. The pesticide was detected in 14 samples among the 3,632 samples tested. Amount of the uniconazole pesticide ranged from 0.098 to 2.2 mg/kg in the 14 samples. This method was described for the simultaneous determination of uniconazole by using gas chromatography with an electron capture detector (GC-ECD) and mass spectrometry (MS). For evaluating the GC-ECD method, uniconazole was spiked into gyeojachae at a level of 0.05, 0.5 mg/kg. The recoveries of uniconazole with the GC-ECD method ranged from 98.9-109.4%. The results indicate that our method of simultaneous analysis is applicable to uniconazole analysis.

Effect of roasting degree of barley on aroma characteristics of boricha (보리의 로스팅 정도가 보리차의 향 특성에 미치는 영향)

  • Joung, Woo-Young;Kim, Su-Jeong;Kim, Byeong-Goo;Hurh, Byung-Serk;Baek, Hyung-Hee
    • Korean Journal of Food Science and Technology
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    • v.50 no.2
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    • pp.123-131
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    • 2018
  • The objective of this study was to evaluate the effect of roasting degree of barley on aroma characteristics of boricha (barley tea) using solvent-assisted flavor evaporation-gas chromatography-mass spectrometry and gas chromatography-olfactometry. Fifteen volatile compounds including pyrazine, ethylpyrazine, butyrolactone, and guaiacol were considered important volatile compounds, which are generated by roasting barley, because concentrations of those volatiles were significantly increased (p<0.05) as roasting degree of barley was darker. Guaiacol (smoky), furfuryl alcohol (burnt sugar), and furfural (caramel) were detected as aroma-active compounds of boricha with high intensity. Aroma intensity of aroma-active compounds in boricha increased with increase in roasting degree of barley. However, one unknown compound with burnt smell was detected as off-flavor in dark roasted barley. Therefore, it is implied that medium roasting of barley is desirable during boricha manufacturing. Furthermore, boricha manufactured with steamed barley contained more abundant volatile flavor compounds, which may lead to better aroma quality of boricha.

Flavor Components in Dried Fruit of the Chinese Matrimony Vine during Storage (저장 기간이 짧은 건조 구기자와 오래된 건조 구기자의 휘발성 성분)

  • Choi, Sung-Hee
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1325-1329
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    • 2014
  • Gugija (Lycii chinese Miller) is traditionally consumed as a Chinese medicinal material in food, tea, or alcoholic beverages. Gugija has beneficial healthy components, but it produces an off-flavor during storage. This study compared the flavor components of fresh-dried Gugija and stale-dried Gugija. The flavor compounds in one fresh sample (sample 1) and one stale sample (sample 2) were extracted by the simultaneous distillation and extraction method. The concentrated aroma extracts were analyzed and identified by gas chromatography-mass spectrometry. Forty-five compounds, including 17 aldehydes, 8 alcohols, 6 terpene compounds, 4 esters, 3 ketones, and 3 pyrazines, were isolated in sample 1. Thirty-four compounds, including 12 aldehydes, 3 alcohols, 5 terpene compounds, 2 esters, 3 ketones, 3 pyrazines, and 1 acid, were isolated in sample 2. The main aroma components of sample 1 were 2-methyl butanal, 2-methyl propanol having sweet odor, and hexanal, (Z)-3-hexenol having grass odor, and phenyl acetaldehyde, benzyl alcohol having floral odor, and alkyl pyrazines having nutty odor. These compounds were decreased in sample 2, and several compounds containing isovaleric acid, which has a disagreeable, rancid-cheese odor were found newley.

Functional and Volatile Flavor Components in Myungtae(Alaska pollack) sikhae (마른명태 식해의 향기성분과 기능성)

  • Koo, Tae-Ho;Zhang, Yun-Bin;Choi, Hee-Jin;Woo, Hi-Seoh;Son, Gyu-Mok;Choi, Cheong
    • Journal of the Korean Society of Food Culture
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    • v.17 no.5
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    • pp.535-542
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    • 2002
  • The volatile compounds of Myungtae (Alaska pollack) sikhae obtained by simultaneous steam distillation and extraction(SDE) apparatus were separated by gas chromatography(GC) and gas chromatography mass spectrometry(GC/MS). The totals of 155 volatile flavor components was identified in traditional Kyungsangdo Myungtae (Alaska pollack) sikhae, respectively. ${\alpha}$-Zingihirene(11.03%) (E)-di-2-propenyl disulfide(7.95%) ${\beta}$-cironellol(6.02%), methyl allyl disulfide(3.58%), cryptone(3.39%), camphene(3.23%), pentanol(3.21%), penadecanal(2.66%) and ${\beta}$-phellandrene(2.06%) were contained as the main compounds of Myungtae shikae. The fraction obtained from sikhae were tested for electron donating ability, angiotensin converting enzyme and xanthine oxidase inhibitory activity. There was no electron donating abilities$(SC_{50})$ of hexane and water fraction. On the other hand, the abilities of ethylacetate fraction and butanol fraction showed $310.64\;{\mu}g/mL,\;1096.49\;{\mu}g/mL$, respectively. Angiotensin converting enzyme inhibitory activities$(IC_{50})$ of ethylacetate fraction and butanol fraction were 1.623 mg/mL, 1.303 mg/mL, respectively. Xanthine oxidase inhibitory activities$(IC_{50})$ of ethylacetate fraction and butanol fraction were 3.591 mg/mL, 2.083 mg/mL, respectively.

Changes of Saponin during the Cultivation of Soybean Sprout (콩나물 생장 중 사포닌의 변화)

  • Oh, Bong-Yun;Park, Bock-Hee;Ham, Kyung-Sik
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1039-1044
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    • 2003
  • We investigated the changes in saponins during the cultivation of soybean sprout. Crude saponin content was 4.15mg/g in germinated soybean and reached its peark (5.33mg/g) in soybean sprout cultivated for six days. Saponin content in the cotyledon, stem, and root of the soybean sprout cultivated for six days were 4.17, 7.46, and 7.45mg/g, respectively. Soyasaponins extracted from the soybean sprout were analyzed with LC-electrospray ionization (ESI)-mass spectrometry, in which a reverse phase $C_18$ column was used for separation of saponins. In the soybeen sprout, group B saponin, I, II, III, IV, and V increased 7, 2, 1.4, 8.7, and 3.3 fold, respectively, compared to those in the soybean seed. Group B saponin I, II, III, IV, and V in the stem of the soybean sprout were 10.53, 1.45, 10.49, 5.72 and 8.14 fold the level of those in the cotyldon, respectively. In the root, the contents of group B saponin I, III, IV, and V were 5.54, 2.77, 4.86 and 9.73 fold, respectively, higher than those in cotyledon, but the content of group B saponin 2 was 2.96 fold less than that in cotyledon. These results indicate that the biosyntheses of group B saponins are differentially regulated in growing soybean sprout.

Nonvolatile Flavor Components in Chinese Quince Fruits, Chaenomeles sinensis koehne (모과의 비휘발성 Flavor 성분에 관한 연구)

  • Chung, Tae-Yung;Cho, Dae-Sun;Song, Jae-Chul
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.293-302
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    • 1988
  • This study was performed to identify detailed informations on the nonvolatile flavor of Chinese quince fruits, Chaenomeles sinensis koehne. About 72% of the free amino acids were shown to be valine, asparagine, ${\gamma}-aminobutyric\;acid$, aspartic acid and serine. Arginine, tyrosine, methionine and tryptophan were not present. Glutamic acid and glutamine as a amino acid for peptides were the major components, whereas cysteic acid, methionine sulfone and tryptophan were not detected. The nucleotides attained were composed of cytosine, uridine-5'-monophosphate and cytidine-5'-monophosphate, and these were proved to be a very small quantity. Guanosine-5'-monophosphate, inosine-5'-monophosphate and adenosine-5'-monophosphate were not present. The major sugars were shown to be glucose, sorbose, sucrose and fructose. Fructose was the most abundant one among them. A total of 11 organic acids were identified by capillary gas chromatography and capillary gas chromatography-mass spectrometry. The major components identified were tartaric acid and α-ketoglutaric acid. The total content of vitamin C determined was 386.6mg%, and those of ascorbic, dehydroascorbic, and 2, 3-diketo-L-gulonic acid were 28.8mg%, 154.5mg% and 197.3mg%, respectively. Calcium and phosphorus were the major components, while heavy metals such as cadmium, copper and lead were determined to be a small amount. In the result of organoleptic test on the natural and synthetic extract of Chinese quince fruits, the principal taste components consisted of free amino acids, sugars, organic acids, vitamin C and minerals. Five groups mentioned would have a favorable influence upon the taste of fresh Chinese quince fruits.

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Chemical Composition and Protective Effect of Essential Oils Derived from Medicinal Plant on PC12 Neuro-cells Induced by Oxidative Stress (약용식물 유래 정유성분 분석 및 산화 스트레스로부터 PC12 신경세포 보호 효과)

  • Lee, Ji Yeon;Park, Jeong-Yong;Kim, Dong Hwi;Choi, Su Ji;Jang, Gwi Young;Seo, Kyung Hye
    • The Korean Journal of Food And Nutrition
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    • v.33 no.2
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    • pp.215-221
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    • 2020
  • The purpose of this study was to investigate the protective effect on oxidative stress induced PC12 cells, and volatile flavor composition of essential oils derived from medicinal plant seeds- Gossypium hirsutum L. (G. hirsutum), Coix lachryma-jobi (C. lachryma-jobi) and Oenothera biennis (O. biennis). The essential oils were obtained by the solvent (hexane) extraction method from the seeds. The essential oils of the seeds were analyzed by the solid-phase micro-extraction gas chromatography mass spectrometry (SPME-GC/MS). The major compounds of G. hirsutum, C. lachryma-jobi and O. biennis were cyclonexanol (16.65%), β-asarone (14.29%) and ylangene (50.01%). The DPPH radical scavenging activity (IC50) was the highest value of 8.52 mg/mL in the O. biennis. Additionally, IC50 values of G. hirsutum and C. lachryma-jobi were 26.76 mg/mL and 36.81 mg/mL. For the oxidative stress on PC12 cells, we treated with hydrogen peroxide (H2O2). The pretreatment of oxidative stress induced PC12 cells with all the essential oils preserved or increased their cell viability and G. hirsutum and O. biennis attenuated the ROS generation (by 68.75% and 56.25% vs. H2O2 control). The results of this study suggest that the essential oils derived from medicinal plant seeds could be used as valuable back data as a natural essential oil material to prevent neurodegenerative diseases by protecting neuro-cells.

Isolation of Flavonoids and Lignans from the Stem Wood of Lindera obtusiloba Blume (생강나무(Lindera obtusiloba Blume) 목부로부터 Flavonoid 및 Lignan 화합물의 분리)

  • Seo, Kyeong-Hwa;Baek, Mi-Young;Lee, Dae-Young;Cho, Jin-Gyeong;Kang, Hee-Cheol;Ahn, Eun-Mi;Baek, Nam-In;Lee, Youn-Hyung
    • Journal of Applied Biological Chemistry
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    • v.54 no.3
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    • pp.178-183
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    • 2011
  • The stem woods of Lindera obtusiloba Blume were extracted in 80% aqueous methanol and the concentrated extract was partitioned with ethyl acetate (EtOAc), butanol (n-BuOH), and $H_2O$, successively. From the EtOAc and n-BuOH fractions, five compounds were isolated through the repeated silica gel, octadecyl silica gel, and Sephadex LH-20 column chromatographies. On the basis of spectroscopic data including mass spectrometry, IR, $^1H$-NMR, $^{13}C$-NMR, distortionless enhancement by polarization transfer, and two-dimensional-NMR gradient correlated spectroscopy (gCOSY), gradient heteronuclear single quantum correlation (gHSQC), gradient heteronuclear multiple bonding connectivity (gHMBC), the chemical structures of the compounds were determined as asarinin (1), (+)-catechin (2), (-)-epicatechin (3), hyperin (4), and nudiposide (5). Compounds 1 and 5 were isolated for the first time from the stem wood of L. obtusiloba Blume.