• 제목/요약/키워드: intracellular uptake

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FABP3 and FABP4 Genes Are the Potential Candidates for Body Weights in Korean Native Chicken

  • Cahyadi, Muhammad;Seo, Dongwon;Choi, Nuri;Jin, Shil;Maharani, Dyah;Heo, Kang Nyeong;Kang, Bo Seok;Jo, Cheorun;Lee, Jun Heon
    • 한국가금학회지
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    • 제40권2호
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    • pp.91-96
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    • 2013
  • FABPs, 15 kDa organic substances, are small intracellular proteins which have a primary role to regulate fatty acid uptake and intracellular transport. This study was conducted to identify SNPs in the two FABP family genes and their associations with the body weight traits in Korean native chicken (KNC). Two SNPs, namely g.508C>T of FABP3 gene and g.285C>T of FABP4 gene, have been genotyped by using PCR-RFLP method. The results showed that FABP3 was significantly associated with body weight at birth, body weights at 12 to 20 weeks, and also slaughter weight. Moreover, the g.285C>T SNP of FABP4 gene was not associated with any body weight traits. These results suggested that the g.508C>T SNP of FABP3 genes can be used as molecular markers to select KNC having desirable body weights.

Thyroid Hormone-Induced Alterations of $Ca^{2+}-ATPase$ and Phospholamban Protein Expression in Cardiac Sarcoplasmic Reticulum

  • Kim, Hae-Won;Noh, Kyung-Min;Park, Mi-Young;Lee, Hee-Ran;Lee, Eun-Hee
    • The Korean Journal of Physiology and Pharmacology
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    • 제3권2호
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    • pp.223-230
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    • 1999
  • Alterations of cardiovascular function associated with various thyroid states have been studied. In hyperthyroidism left ventricular contractility and relaxation velocity were increased, whereas these parameters were decreased in hypothyroidism. The mechanisms for these changes have been suggested to include alterations in the expression and/or activity levels of various proteins; ${\alpha}-myosin$ heavy chain, ${\beta}-myosin$ heavy chain, ${\beta}-receptors,$ the guanine nucleotide-binding regulatory protein, and the sarcolemmal $Ca^{2+}-ATPase.$ All these cellular alterations may be associated with changes in the intracellular $Ca^{2+}$ concentration. The most important regulator of intracellular $Ca^{2+}$ concentration is the sarcoplasmic reticulum (SR), which serves as a $Ca^{2+}$ sink during relaxation and as a $Ca^{2+}$ source during contraction. The $Ca^{2+}-ATPase$ and phospholamban are the most important proteins in the SR membrane for muscle relaxation. The dephosphorylated phospholamban inhibits the SR $Ca^{2+}-ATPase$ through a direct interaction, and phosphorylation of phospholamban relieves the inhibition. In the present study, quantitative changes of $Ca^{2+}-ATPase$ and phospholamban expression and the functional consequences of these changes in various thyroid states were investigated. The effects of thyroid hormones on (1) SR $Ca^{2+}$ uptake, (2) phosphorylation levels of phospholamban, (3) SR $Ca^{2+}-ATPase$ and phospholamban protein levels, (4) phospholamban mRNA levels were examined. Our findings indicate that hyperthyroidism is associated with increases in $Ca^{2+}-ATPase$ and decreases in phospholamban levels whereas opposite changes in these proteins occur in hypothyroidism.

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Investigation into the Efficacy of Val-SN-38, a Valine-Ester Prodrug of the Anti-Cancer Agent SN-38

  • Kwak, Eun-Young;Choi, Min-Koo;Yang, Su-Geun;Shim, Chang-Koo;Shim, Won-Sik
    • Biomolecules & Therapeutics
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    • 제20권3호
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    • pp.326-331
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    • 2012
  • We recently reported that Val-SN-38, a novel valine ester prodrug of SN-38, had greatly improved the intracellular accumulation of SN-38 in MCF-7 cell line, probably through enhanced uptake via amino acid transporters. In the present study, the efficacy of Val-SN-38 was further investigated both in vitro and in vivo. It was found that the in vitro cytotoxic effect of Val-SN-38 was similar to that of SN-38. Moreover, Val-SN-38 exhibited an equal potency to that of SN-38 in survival experiments in vivo. Because these results seemed to be contrary to the previous finding, further investigation was performed to find out the underlying cause of the contradiction. As only the lactone form is known to have cytotoxic activity, the proportion of lactone in Val-SN-38 and SN-38 was determined, but no differences were found. However, it turned out that Val-SN-38 had poor stability compared with SN-38, which resulted in a decrease in beneficial efficacy for Val-SN-38. Overall, the present study showed that a valine-added prodrug approach could be advantageous provided that the stability of the compound can be ensured. We believe this is a noteworthy study that unravels the discrepancy between intracellular accumulation and efficacy of valine-added prodrug.

Protection of palmitic acid treatment in RAW264.7 cells and BALB/c mice during Brucella abortus 544 infection

  • Reyes, Alisha Wehdnesday Bernardo;Huy, Tran Xuan Ngoc;Vu, Son Hai;Kim, Hyun Jin;Lee, Jin Ju;Choi, Jeong Soo;Lee, John Hwa;Kim, Suk
    • Journal of Veterinary Science
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    • 제22권2호
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    • pp.18.1-18.12
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    • 2021
  • Background: We previously elucidated the protective mechanism of Korean red ginseng oil (RGO) against Brucella abortus infection, and our phytochemical analysis revealed that palmitic acid (PA) was an abundant component of RGO. Consequently, we investigated the contribution of PA against B. abortus. Objectives: We aimed to investigate the efficacy of PA against B. abortus infection using a murine cell line and a murine model. Methods: Cell viability, bactericidal, internalization, and intracellular replication, western blot, nitric oxide (NO), and superoxide (O2-) analyses and flow cytometry were performed to determine the effects of PA on the progression of B. abortus infection in macrophages. Flow cytometry for cytokine analysis of serum samples and bacterial counts from the spleens were performed to determine the effect of PA in a mouse model. Results: PA did not affect the growth of B. abortus. PA treatment in macrophages did not change B. abortus uptake but it did attenuate the intracellular survivability of B. abortus. Incubation of cells with PA resulted in a modest increase in sirtuin 1 (SIRT1) expression. Compared to control cells, reduced nitrite accumulation, augmented O2-, and enhanced pro-inflammatory cytokine production were observed in PA-treated B. abortus-infected cells. Mice orally treated with PA displayed a decreased serum interleukin-10 level and enhanced bacterial resistance. Conclusions: Our results suggest that PA participates in the control of B. abortus within murine macrophages, and the in vivo study results confirm its efficacy against the infection. However, further investigations are encouraged to completely characterize the mechanisms involved in the inhibition of B. abortus infection by fatty acids.

Protective Effects against Brucella abortus 544 Infection in a Murine Macrophage Cell Line and in a Mouse Model via Treatment with Sirtuin 1 Activators Resveratrol, Piceatannol and Ginsenoside Rg3

  • Alisha Wehdnesday Bernardo Reyes;Heejin Kim;Tran Xuan Ngoc Huy;Trang Thi Nguyen;Wongi Min;Hu Jang Lee;Jin Hur;John Hwa Lee;Suk Kim
    • Journal of Microbiology and Biotechnology
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    • 제33권4호
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    • pp.441-448
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    • 2023
  • Brucellosis is a contagious zoonotic disease that infects millions of people annually with hundreds of millions more being exposed. It is caused by Brucella, a highly infectious bacterial species capable of infecting humans with an estimated dose of 10-100 organisms. Sirtuin 1 (SIRT1) has been reported to contribute to prevention of viral diseases as well as a chronic infection caused by Mycobacterium bovis. Here, we investigated the role of SIRT1 in the establishment of Brucella abortus infection in both in vitro and in vivo systems using the reported SIRT1 activators resveratrol (RES), piceatannol (PIC), and ginsenoside Rg3 (Rg3). In RAW264.7 cells, SIRT1 activators did not alter the adherence of Brucella or Salmonella Typhimurium. However, reduced uptake of Brucella was observed in cells treated with PIC and Rg3, and survival of Brucella within the cells was only observed to decrease in cells that were treated with Rg3, while PIC treatment reduced the intracellular survival of Salmonella. SIRT1 treatment in mice via oral route resulted in augmented Brucella resistance for PIC and Rg3, but not RES. PIC treatment favors Th2 immune response despite reduced serum pro-inflammatory cytokine production, while Rg3-treated mice displayed high IL-12 and IFN-γ serum production. Overall, our findings encourage further investigation into the complete mechanisms of action of the different SIRT1 activators used as well as their potential benefit as an effective alternative approach against intracellular and extracellular pathogens.

칼슘 길항제가 활성화된 호중구에서의 $O_{\overline{2}}$의 생성, NADPH oxidase활성도 및 탐식작용에 미치는 영향 (Effects of Calcium Antagonists on Superoxide Generation, NADPH Oxidase Activity and Phagocytic Activity in Activated Neutrophils)

  • 이정수;한은숙;이광수
    • 대한약리학회지
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    • 제23권1호
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    • pp.33-44
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    • 1987
  • 면역보체가 결합되어 있는 zymosan 또는 열로 응집된 IgG에 의하여 활성화된 호중구에서의 NADPH oxidase 의존적인 $O_{\overline{2}}$의 생성과 탐식작용은 칼슘의 흡수과정과 일치하였다. 활성화된 백혈구의 반응은 세포외 칼슘 농도에 따라 항진되었으며, 이는 칼슘의 킬레이트제인 EGTA나 EDTA에 의하여 유의하게 억제되었다. 활성화된 백혈구로부터 $O_{\overline{2}}$의 생성은 dantrolene과 chlorpromazine에 의하여 억제되었다. 칼슘 길항제인 bepredil, diltiazem, verapamil, nifedipine, nimodipine은 효과적으로 활성화된 백혈구의 칼슘흡수, $O_{\overline{2}}$ 생성 그리고 탐식 작용을 억제하였고 NADPH oxidase 활성도 또한 억제하였다. 그러므로, 칼슘 길항제는 칼슘 유입을 억제하거나 칼슘의 세포내 재분포 및 NADPH oxidase 반응계에 작용하여 활성화된 백혈구에서의 $O_{\overline{2}}$의 생성과 백혈구의 탐식작용을 억제할 것으로 시사되었다.

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Effects of L-trans-pyrrolidine-2,4-dicarboxylate, a Glutamate Uptake Inhibitor, on NMDA Receptor-mediated Calcium Influx and Extracellular Glutamate Accumulation in Cultured Cerebellar Granule Neurons

  • Oh, Seikwan;Shin, Chang-Sik;Patrick-P. McCaslin;Seong, Yeon-Hee;Kim, Hack-Seang
    • Archives of Pharmacal Research
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    • 제20권1호
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    • pp.7-12
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    • 1997
  • Glutamate uptake inhibitor, L-trans-pyrrolidine-2, 4-dicarboxylate (PDC, $20{\mu}M$) elevated basal and N-methyl-D-aspartate (NMDA, $100{\mu}M$)-induced extracellular glutamate accumulation, while it did not augment kainate $100{\mu}M$-induced glutamate accumulation in cultured cerebellar granule neurons. However, pretreatment with PDC for 1 h significantly reduced NMDA-induced glutamate accumulation, but did not affect kainate-induced response. Pretreatment with glutamate $(5{\mu}M)$ for 1 h also reduced NMDA-induced glutamate accumulation, but did not kainate-induced response. Upon a brief application (3-10 min), PDC did neither induce elevation of intracellular calcium concentration $([Ca^{2+}]_i)$ nor modulate NMDA-indLiced $[Ca^{2+}]_1$ elevation. Pretreatment with PDC for 1 h reduced NMDA-induced $[Ca^{2+}]_1$ elevation, but it did not reduce kainate-induced $[Ca^{2+}]_1$ elevation. These results suggest that glutamate concentration in synaptic clefts of neurana cells is increased by prolonged exposure (1 h) of the cells to PDC, and the accumulated glutamate subsequently induces selective desensitization of NMDA receptor.

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Tc-99m DPD 골스캔과 복부 CT 영상에서 보이는 위암의 석회화 (Visualization of the Gastric Calcification due to Cancer on Tc-99m DPD and Abdominal CT Images)

  • 정영진;강도영
    • 대한핵의학회지
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    • 제38권5호
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    • pp.344-346
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    • 2004
  • A 69-year-old woman was presented with progressed dysphagia, gastric soreness and weight loss during 2 months. She was performed abdomen x-ray, EGDS and abdomen CT. Abdomen x-ray demonstrated punctuate calcification on LUQ. EGDS showed an ulceroinfiltrative mass with bleeding on cardia to antrum of stomach. And CT showed diffuse gastric wall thickness with multiple calcifications. Biopsy of the stomach and esophagus during EGDS examination revealed an adenocarcinoma, with signet ring cell type, infiltrating the wall of the stomach and the distal esophagus. Then acne scan was performed a few days later. It revealed intense uptake in LUQ, corresponding to the calcium containing neoplasm seen on the abdomen x-ray, EGDS and abdomen CT. And there was no evidence of any metastatic lesion and thyroid uptake on the bone scan. There are many reports about accumulation of the tracer in extraosseous lesion, but only a few literatures were reported about gastric calcification in stomach cancer. More over, no reports showed CT images. We are performed many diagnostic examinations and found well correlation between them. The reason of gastric calcification is considered with calcium deposition within extracellular space due to hemorrhage or necrosis. Other possibility offered to explain gastric calcification have been increased blood flow and/or increased neovascularity with capillary leaks of tracer, and specific enzymatic (phosphatases) receptor binding of tracer. So, it was happened ion exchange between intracellular calcium and phosphate groups of tracer.

Minimal Amount of Insulin Can Reverse Diabetic Heart Function: Sarcoplasmic Reticulum $Ca^{2+}$ Transport and Phospholamban Protein Expression

  • Kim, Hae-Won;Cho, Yong-Sun;Lee, Yun-Song;Lee, Eun-Hee;Lee, Hee-Ran
    • The Korean Journal of Physiology and Pharmacology
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    • 제3권2호
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    • pp.175-182
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    • 1999
  • In the present study, the underlying mechanisms for diabetic functional derangement and insulin effect on diabetic cardiomyopathy were investigated with respect to sarcoplasmic reticulum (SR) $Ca^{2+}-ATPase$ and phospholamban at the transcriptional and translational levels. The maximal $Ca^{2+}$ uptake and the affinity of $Ca^{2+}-ATPase$ for $Ca^{2+}$ were decreased in streptozotocin-induced diabetic rat cardiac SR, however, even minimal amount of insulin could reverse both parameters. Levels of both mRNA and protein of phospholamban were significantly increased in diabetic rat hearts, whereas the mRNA and protein levels of SR $Ca^{2+}-ATPase$ were significantly decreased. In case of phospholamban, insulin treatment reverses these parameters to normal levels. Minimal amount of insulin could reverse the protein levels; however, it could not reverse the mRNA level of SR $Ca^{2+}-ATPase$ at all. Thus, the decreased SR $Ca^{2+}$ uptake appear to be largely attributed to the decreased SR $Ca^{2+}-ATPase$ level, which is further impaired due to the inhibition by the increased level of phospholamban. These results indicate that insulin is involved in the control of intracellular $Ca^{2+}$ in the cardiomyocyte through multiple target proteins via multiple mechanisms for the decrease in the mRNA for both SR $Ca^{2+}-ATPase$ and phospholamban which are unknown and needs further study.

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HeLa세포에서 IgE-dependent Histamine-releasing Factor의 인산화가 Na,K-ATPase의 활성에 미치는 영향 (Studies on the Effect of the Phosphorylated IgE-Dependent Histamine-Releasing Factor on Na,K-ATPase Activity in HeLa Cell)

  • 김정아;하헌주;이경림
    • 한국미생물·생명공학회지
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    • 제33권3호
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    • pp.184-188
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    • 2005
  • HRF는 Na,K-ATPase의 ${\alpha}$ subunit에 결합하여 이의 활성을 저해하는 것으로 알려져 있으며, PKC에 의해 Ser98 잔기가 인산화 될 수 있다는 것을 anti-HRFpS98 항체와 HRF S98A mutant를 이용한 실험으로 확인할 수 있었다. 또한 $^{86}Rb^{+}-uptake$ assay 실험에서 HRF의 serine 98 잔기의 탈인산화는 Na,K-ATPase의 활성에 약간의 영향을 미치는 것으로 미루어 PKC에 의해 인산화되는 98 serine 잔기가 Na,K-ATPase 활성 저해에 큰 영향을 미치지 않는 것으로 보인다.