• Title/Summary/Keyword: P-glycoprotein 2

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Cloning and Sequencing of a Gene Cluster for the Resistance to Doxorubicin from Streptomyces peucetius subsp. caesius ATCC 27952

  • Hong, Young-Soo;Hwang, Cheol-Kyu;Hwang, Dong-Youn;Kim, Young-Ho;Kim, Sung-Jun;Lee, Jung-Joon
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.153-160
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    • 1992
  • The doxorubicin resistance locus from Streptomyces peucetius subsp. caesius (the doxorubicin producer, ATCC 27952) has been cloned. The sequence data over 4.4 kb regions reveals the presence of four possible open reading frames (ORFs). ORF2 and ORF3 would encode proteins containing 329 and 283 amino acids, respectively. The protein encoded by ORF2 has two almost identical ATP binding domains with p-glycoprotein, the product of a multidrug resistance gene from tumor cells, and that encoded by ORF3 has several hydrophobic domains suggesting that it is located in the bacterial membrane. These two remarkable similarities of the gene product to p-glycoprotein of mammalian tumor cells suggest that the two proteins may enable bacteria to extrude a variety of toxic agents, including daunorubicin and doxorubicin, by an ATP dependent efflux mechanism analogous to the multidurg resistance protein of cancer cells.

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Killer 효모 융합주 FWKS 260 이 분비하는 Killer Toxin 의 정제

  • 정기택;방광웅;우철주;정용진;김재근;송형익
    • Korean Journal of Microbiology
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    • v.30 no.3
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    • pp.160-163
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    • 1992
  • Killer toxin from killer yeast fusant FWKS 260 developed by protoplast fusion between the wild killer yeast and alcohol-fermenting yeast was purified by ammonium sulfate fractionation. Amicon PM I0 concentration. Sephadex G-200 and Scphadcx G-75 column chromatography. The purified killer toxin showed a single band by SIX-polyacvlamide gel electrophoresis. The protein part of killer toxin was active site. which was found by treating the proteolytic enzyme such as pronase E and pepsin to killer toxin. The killer toxin was stable at pH 2.0-5.0 and 20$^{\circ}$C. but inactivated with increasing temperature. The molecular weight was determined to be approximately 13.000 according to the results obtained from the SDS-polyacrylamide gel electrophoresis. It was confirmed that the purified killer toxin is glycoprotein by showing a red single band after st'tining with Schiffs reagent.

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Inhibitory effect of ethanol extract of Gryllus bimaculatus on platelet aggregation and glycoprotein IIb/IIIa activation (쌍별귀뚜라미 에탄올 추출물의 혈소판응집반응과 당단백질 IIb/IIIa 활성화 억제 효과)

  • Hyuk-Woo Kwon;Man Hee Rhee;Jung-Hae Shin
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.236-243
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    • 2023
  • Platelets act a fundamental role in primary- and secondary-hemostasis, however, platelet activation may cause thrombosis simultaneously. Therefore, control of platelet aggregation is crucial in preventing thrombosis-mediated diseases. Recently, the development of insect materials is attracting attention. Among the highly nutritious functional food sources, insects such as two-spotted cricket (Gryllus bimaculatus). Gryllus bimaculatus (G. bimaculatus) contains high protein and unsaturated fatty acids and has been registered as a food material September 2015 by the Ministry of Food and Drug Safety of Korea. In this study, we examined whether G. bimaculatus extract (GBE) inhibits platelet aggregation, intracellular calcium mobilization, thromboxane A2 production and glycoprotein IIb/IIIa (integrin αIIb/β3) activation. We investigated whether GBE can regulate signaling molecules, such as 1, 4, 5-triphosphate receptor type I, extracellular signal-regulated kinase, cytosolic phospholipase A2, mitogen-activated protein kinases p38, vasodilator-stimulated phosphoprotein, phosphatidylinositol-3 kinase, Akt, glycogen synthase kinase-3α/β, and SYK. Taken together, GBE is a potential therapeutic drug candidate to prevent platelet-related thrombosis and cardiovascular disease.

Effects of Amlodipine on the Pharmacokinetics of Warfarin after Oral and Intravenous Administration of Warfarin in Rats

  • Choi, Dong-Hyun;Piao, Yong-Ji;Choi, Eun-Joo;Choi, Jun-Shik;Burm, Jin-Pil
    • Biomolecules & Therapeutics
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    • v.19 no.4
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    • pp.493-497
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    • 2011
  • The aim of this study was to investigate the effect of amlodipine on the pharmacokinetics of warfarin after oral and intravenous administration of warfarin in rats. Warfarin was administered orally (0.2 mg/kg) or intravenously (0.05 mg/kg) without or with oral administration of amlodipine (0.1 or 0.4 mg/kg) in rats. The effect of amlodipine on the P-glycoprotein (P-gp) as well as cytochrome P450 (CYP) 3A4 activity was also evaluated. Amlodipine inhibited CYP3A4 enzyme activity with 50% inhibition concentration ($IC_{50}$) of 9.1 ${\mu}M$. Compared to those animals in the oral control group (warfarin without amlodipine), the area under the plasma concentration-time curve (AUC) of warfarin was significantly greater (0.1 mg/kg, p<0.05; 0.4 mg/kg, p<0.01) by 26.5-53.5%, and the peak plasma concentration ($C_{max}$) was significantly higher (0.4 mg/kg, p<0.05) by 26.2% after oral administration of warfarin with amlodipine, respectively. Consequently, the relative bioavailability of warfarin increased by 1.26- to 1.53-fold and the absolute bioavailability of warfarin with amlodipine was significantly greater by 61.7-72.5% compared to that in the control group (47.4%). In contrast, amlodipine had no effect on any pharmacokinetic parameters of warfarin given intravenously. Therefore, the enhanced oral bioavailability of warfarin may be due to inhibition of CYP 3A4-mediated metabolism in the intestine and/or liver rather than renal elimination and P-gp by amlodipine.

Purification and Characterization of a Lectin from Arisaema tortuosum Schott Having in-vitro Anticancer Activity against Human Cancer Cell Lines

  • Dhuna, Vikram;Bains, Jagmohan Singh;Kamboj, Sukhdev Singh;Singh, Jatinder;Shanmugavel;Saxena, Ajit Kumar
    • BMB Reports
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    • v.38 no.5
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    • pp.526-532
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    • 2005
  • A lectin with in-vitro anticancer activity against established human cancer cell lines has been purified by affinity chromatography on asialofetuin-linked amino activated silica beads from the tubers of Arisaema tortuosum, popularly known as Himalayan Cobra lily, a monocot plant from the family Araceae. The bound Arisaema tortuosum lectin (ATL) was eluted with glycine-HCl buffer, pH 2.5. ATL was effectively inhibited by asialofetuin, a complex desialylated serum glycoprotein as well as by N-acetyl-D-lactosamine, a disaccharide. It gave a single band corresponding to a subunit molecular weight of 13.5 kDa in SDS-PAGE, pH 8.8 both under reducing and non reducing conditions. When subjected to gel-filtration on Biogel P-200, it was found to have a molecular weight of 54 kDa, suggesting a homotetramer structure, in which individual polypeptides are not bound to each other with disulfide bonds. ATL is a glycoprotein with 0.9% carbohydrate content, stable up to $55^{\circ}C$ and at pH 2 to 10. The lectin had no requirement for divalent metal ions i.e. $Ca^{2+}$ and $Mn^{2+}$ for its activity. However, as reported for other monocot lectins, ATL gave multiple bands in isoelectric focusing and Native PAGE, pH 8.3. The lectin was found to inhibit in vitro proliferation of human cancer cell lines HT29, SiHa and OVCAR-5.

Expression Levels of Plasma Angiogenic Factors during Early Pregnancy in Hanwoo

  • Bae, Seong-Hun;Yang, Byoung-Chul;Joo, Seok-Cheon;Min, Kwan-Sik;Yoon, Jong-Taek;Jin, Dong-Il;Kim, Tae-Hun;Seong, Hwan-Hoo;Hwang, Seong-Soo
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.263-267
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    • 2008
  • This study was conducted to compare the expression pattern of the specific factors associated with pregnancy and angiogenesis during early pregnancy in Hanwoo. Synchronized female Hanwoo ($4{\sim}6$ year-old) were inseminated artificially. After 10 weeks after artificial insemination (AI), the pregnancy was tested by rectal palpation method. Three pregnant and non-pregnant Hanwoo were used in this experiment, respectively. The plasma progesterone level was measured by ELISA. Western blot analysis was performed to detect the expression of pregnancy associated glycoprotein (PAG) or angiogenic factors (VEGF, B-FGF, ANP-1, and TIE-2). The plasma P4 level was increase gradually in pregnant group and maintained high level. The concentration of PAG was significantly higher from $5^{th}$ weeks in pregnant group compared to that of non-pregnant group (p<0.05). The concentrations of the VEGF (p<0.05), B-FGF (p<0.05), and ANP-1 (p<0.05) were significantly increased from $6^{th}\;or\;7^{th}$ week after AI in pregnant group, respectively. And the intensity of TIE-2, ANP-1 receptor, was well matched with ANP-1 (p<0.05). Taken together, it can be postulated that the blood vessels connected with fetus and dam were formed dramatically around 40 days after AI, because the expression levels of the angiogenic factors were increased significantly from this time in pregnant Hanwoo.

Effects of Curcumin on the Pharmacokinetics of Loratadine in Rats: Possible Role of CYP3A4 and P-glycoprotein Inhibition by Curcumin

  • Li, Cheng;Choi, Byung-Chul;Kim, Dong-Ki;Choi, Jun-Shik
    • Biomolecules & Therapeutics
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    • v.19 no.3
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    • pp.364-370
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    • 2011
  • The purpose of this study was to investigate the effects of curcumin on the pharmacokinetics of loratadine in rats. The effect of curcumin on P-glycoprotein (P-gp) and cytochrome P450 (CYP) 3A4 activity was evaluated. Pharmacokinetic parameters of loratadine were also determined after oral and intravenous administration in the presence or absence of curcumin. Curcumin inhibited CYP3A4 activity with an IC50 value of 2.71 ${\mu}M$ and the relative cellular uptake of rhodamine-123 was comparable. Compared to the oral control group, curcumin significantly increased the area under the plasma concentration-time curve and the peak plasma concentration by 39.4-66.7% and 34.2-61.5%. Curcumin also significantly increased the absolute bioavailability of loratadine by 40.0-66.1% compared to the oral control group. Consequently, the relative bioavailability of loratadine was increased by 1.39- to 1.67-fold. In contrast, curcumin had no effect on any pharmacokinetic parameters of loratadine given intravenously, implying that the enhanced oral bioavailability may be mainly due to increased intestinal absorption caused via P-gp and CYP3A4 inhibition by curcumin rather than to reduced renal and hepatic elimination of loratadine. Curcumin enhanced the oral bioavailability of loratadine in this study. The enhanced bioavailability of loratadine might be mainly attributed to enhanced absorption in the gastrointestinal tract via the inhibition of P-gp and reduced fi rst-pass metabolism of loratadine via the inhibition of the CYP3A subfamily in the small intestine and/or in the liver by curcumin.

Sequence Analysis of E2 Glycoprotein from Indian Isolate of Classical Swine Fever Virus (CSFV)

  • Bajwa, Mehak;Verma, Ramneek;Deka, Dipak;Dhol, Gagandeep Singh;Barman, Nagendra Nath
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.22-30
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    • 2015
  • CSF is a major concern for the swine industry, representing currently the most epizootically dangerous disease to the species. Numerous CSFV isolates with various degrees of virulence have already been isolated worldwide, ranging from low virulent strains that do not result in any apparent clinical signs to highly virulent strains that cause a severe per acute hemorrhagic fever with very high mortality. The molecular epidemiology of CSFVs has proven to be an essential tool for effective disease control and the development of safe and effective vaccines. Therefore, this study cloned and sequenced local CSFV isolates, and conducted a phylogenetic analysis based on the E2 glycoprotein encoding sequences.The RNA was extracted from PK15 cell culture passaged CSFV isolates, the cDNA prepared, and the complete E2 gene amplified with a product size of 1186 bp. The gelpurified PCR product was cloned into a pGEMT easy vector and the positive clone commercially sequenced. Aligning the nucleotide (1119 bp) and amino acid (373) sequences with 29 reference strains revealed nucleotide and amino acid sequence identities of 82.60-97.80% and 88.70-98.70%, respectively, indicating a higher mutation rate of the field CSFV strains. The phylogenetic analysis based on the complete E2 amino acid sequences also revealed a reliable differentiation of all the analyzed strains into specific genetic groups and subgroups, plus the local isolate (CSFV-E2) was found to cluster with the CSFV subgroup 2.2. Thus, the full-length E2 cds proved to be most suitable for a reliable and statistically significant phylogenetic analysis of CSFV isolates.

Pharmacokinetic Interaction Between Atorvastatin and Nifedipine (아톨바스타틴과 니페디핀의 약물동태학적 상호작용)

  • Moon, Hong-Seop;Choi, Jun-Shik
    • Korean Journal of Clinical Pharmacy
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    • v.20 no.1
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    • pp.25-29
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    • 2010
  • The purpose of this study was to investigate the effect of atorvastatin on the pharmacokinetics of nifedipine (6 mg/kg) after oral administration of nifedipine with or without atorvastatin (0.5 and 2.0 mg/kg) in rats, and also was to evaluate to the effect of atorvastatin on the CYP3A4 activity. The 50% inhibiting concentration ($IC_{50}$) values of atorvastatin on CYP3A4 activity is 46.1 ${\mu}M$. Atorvastatin inhibited CYP3A4 enzyme activity in a concentration-dependent manner. Coadministration of atorvastatin increased significantly (p<0.05, 2.0 mg/kg) the plasma concentration-time curve (AUC) and the peak concentration ($C_{max}$) of nifedipine compared to the control group. The relative bioavailability (RB%) of nifedipine was increased from 1.15- to 1.37-fold. Coadministration of atorvastatin did not significantly change the terminal half-life ($T_{1/2}$) and the time to reach the peak concentration ($T_{max}$) of nifedipine. Based on these results, we can make a conclusion that the significant changes of these pharmacokinetic parameters might be due to atorvastatin, which possesses the potency to inhibit the metabolizing enzyme (CYP3A4) in the liver and intestinal mucosa, and also inhibit the P-glycoprotein (P-gp) efflux pump in the intestinal mucosa. It might be suggested that atorvastatin altered disposition of nifedipine by inhibition of both the first-pass metabolism and P-glycoprotein efflux pump in the small intestine of rats. In conclusion, the presence of atorvastatin significantly enhanced the oral bioavailability of nifedipine, suggesting that concurrent use of atorvastatin with nifedipine should require close monitoring for potential drug interation.