• 제목/요약/키워드: Small protein

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Modulation of Drug Resistance in Ovarian Cancer Cells by Inhibition of Protein Kinase C-alpha (PKC-α) with Small Interference RNA (siRNA) Agents

  • Zhao, Li-Jun;Xu, Heng;Qu, Jun-Wei;Zhao, Wan-Zhou;Zhao, Yi-Bing;Wang, Jin-Hua
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3631-3636
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    • 2012
  • Objective: To determine whether silence of $PKC-{\alpha}$ expression by small interference RNA (siRNA) might regulate MDR1 expression and reverse chemoresistance of ovarian cancer. Methods: We measured gene and protein expression of MDR1 and $PKC-{\alpha}$ in ovarian cancer cells and assessed their correlation with cell drug resistance. We also examined whether blocking $PKC-{\alpha}$ by RNA interference (RNAi) affected MDR1 expression and reversed drug resistance in drug sensitivity tests. Results: The drug resistance cell lines, OV1228/DDP and OV1228/Taxol, had higher gene and protein expression of MDR1 and $PKC-{\alpha}$ than their counterpart sensitive cell line, OV1228. SiRNA depressed $PKC-{\alpha}$ gene protein expression, as well as MDR1 and protein expression and improved the drug sensitivity in OV1228/DDP and OV1228/Taxol cells. Conclusion: These results indicated that decreasing $PKC-{\alpha}$ expression with siRNA might be an effective method to improve drug sensitivity in drug resistant cells with elevated levels of $PKC-{\alpha}$ and MDR1. A new siRNA-based therapeutic strategy targeting $PKC-{\alpha}$ gene could be designed to overcome the chemoresistance of ovarian cancer.

Manipulation of Cassava Cultivation and Utilization to Improve Protein to Energy Biomass for Livestock Feeding in the Tropics

  • Wanapat, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.463-472
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    • 2003
  • Cassava (Manihot esculenta, Crantz), an annual tropical tuber crop, was nutritionally evaluated as a foliage for ruminants, especially dairy cattle. Cultivation of cassava biomass to produce hay is based on a first harvest of the foliage at three months after planting, followed every two months thereafter until one year. Inter-cropping of leguminous fodder as food-feed between rows of cassava, such as Leucaena leucocephala or cowpea (Vigna unculata), enriches soil fertility and provides additional fodder. Cassava hay contained 20 to 25% crude protein in the dry matter with good profile of amino acids. Feeding trials with cattle revealed high levels of DM intake (3.2% of BW) and high DM digestibility (71%). The hay contains tannin-protein complexes which could act as rumen by - pass protein for digestion in the small intestine. As cassava hay contains condensed tannins, it could have subsequent impact on changing rumen ecology particularly changing rumen microbes population. Therefore, supplementation with cassava hay at 1-2 kg/hd/d to dairy cattle could markedly reduce concentrate requirements, and increase milk yield and composition. Moreover, cassava hay supplementation in dairy cattle could increase milk thiocyanate which could possibly enhance milk quality and milk storage, especially in small holder-dairy farming. Condensed tannins contained in cassava hay have also been shown to potentially reduce gastrointestinal nematodes in ruminants and therefore could act as an anthelmintic agent. Cassava hay is therefore an excellent multi-nutrient source for animals, especially for dairy cattle during the long dry season, and has the potential to increase the productivity and profitability of sustainable livestock production systems in the tropics.

Effect of BIS depletion on HSF1-dependent transcriptional activation in A549 non-small cell lung cancer cells

  • Yun, Hye Hyeon;Baek, Ji-Ye;Seo, Gwanwoo;Kim, Yong Sam;Ko, Jeong-Heon;Lee, Jeong-Hwa
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.4
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    • pp.457-465
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    • 2018
  • The expression of BCL-2 interacting cell death suppressor (BIS), an anti-stress or anti-apoptotic protein, has been shown to be regulated at the transcriptional level by heat shock factor 1 (HSF1) upon various stresses. Recently, HSF1 was also shown to bind to BIS, but the significance of these protein-protein interactions on HSF1 activity has not been fully defined. In the present study, we observed that complete depletion of BIS using a CRISPR/Cas9 system in A549 non-small cell lung cancer did not affect the induction of heat shock protein (HSP) 70 and HSP27 mRNAs under various stress conditions such as heat shock, proteotoxic stress, and oxidative stress. The lack of a functional association of BIS with HSF1 activity was also demonstrated by transient downregulation of BIS by siRNA in A549 and U87 glioblastoma cells. Endogenous BIS mRNA levels were significantly suppressed in BIS knockout (KO) A549 cells compared to BIS wild type (WT) A549 cells at the constitutive and inducible levels. The promoter activities of BIS and HSP70 as well as the degradation rate of BIS mRNA were not influenced by depletion of BIS. In addition, the expression levels of the mutant BIS construct, in which 14 bp were deleted as in BIS-KO A549 cells, were not different from those of the WT BIS construct, indicating that mRNA stability was not the mechanism for autoregulation of BIS. Our results suggested that BIS was not required for HSF1 activity, but was required for its own expression, which involved an HSF1-independent pathway.

Valeriana jatamansi Jones Inhibits Rotavirus-Induced Diarrhea via Phosphatidylinositol 3-Kinase/Protein Kinase B Signaling Pathway

  • Zhang, Bin;Wang, Yan;Jiang, Chunmao;Wu, Caihong;Guo, Guangfu;Chen, Xiaolan;Qiu, Shulei
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1115-1122
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    • 2021
  • Rotavirus (RV), as the main cause of diarrhea in children under 5 years, contributes to various childhood diseases. Valeriana jatamansi Jones is a traditional Chinese herb and possesses antiviral effects. In this study we investigated the potential mechanisms of V. jatamansi Jones in RV-induced diarrhea. MTT assay was performed to evaluate cell proliferation and the diarrhea mice model was constructed using SA11 infection. Mice were administered V. jatamansi Jones and ribavirin. Diarrhea score was used to evaluate the treatment effect. The enzyme-linked immunosorbent assay was performed to detect the level of cytokines. Western blot and quantitative reverse transcription-PCR were used to determine protein and mRNA levels, respectively. Hematoxylin-eosin staining was applied to detect the pathological change of the small intestine. TdT-mediated dUTP nick-end labeling was conducted to determine the apoptosis rate. The results showed V. jatamansi Jones promoted MA104 proliferation. V. jatamansi Jones downregulated phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in protein level, which was consistent with the immunohistochemistry results. Moreover, V. jatamansi Jones combined with ribavirin regulated interleukin-1β (IL-1β), interferon γ, IL-6, tumor necrosis factor α, and IL-10, and suppressed secretory immunoglobulin A secretion to remove viruses and inhibit dehydration. V. jatamansi Jones + ribavirin facilitated the apoptosis of small intestine cells. In conclusion, V. jatamansi Jones may inhibit RV-induced diarrhea through PI3K/AKT signaling pathway, and could therefore be a potential therapy for diarrhea.

Effects of Halophilic Peptide Fusion on Solubility, Stability, and Catalytic Performance of $\small{D}$-Phenylglycine Aminotransferase

  • Javid, Hossein;Jomrit, Juntratip;Chantarasiri, Aiya;Isarangkul, Duangnate;Meevootisom, Vithaya;Wiyakrutta, Suthep
    • Journal of Microbiology and Biotechnology
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    • v.24 no.5
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    • pp.597-604
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    • 2014
  • $\small{D}$-Phenylglycine aminotransferase ($\small{D}$-PhgAT) from Pseudomonas stutzeri ST-201 is useful for enzymatic synthesis of enantiomerically pure $\small{D}$-phenylglycine. However, its low protein solubility prevents its application at high substrate concentration. With an aim to increase the protein solubility, the N-terminus of $\small{D}$-PhgAT was genetically fused with short peptides ($A_1$ ${\alpha}$-helix, $A_2$ ${\alpha}$-helix, and ALAL, which is a hybrid of $A_1$ and $A_2$) from a ferredoxin enzyme of a halophilic archaeon, Halobacterium salinarum. The fused enzymes $A_1$-$\small{D}$-PhgAT, $A_2$-$\small{D}$-PhgAT, and ALAL-$\small{D}$-PhgAT displayed a reduced pI and increased in solubility by 6.1-, 5.3-, and 8.1- fold in TEMP (pH 7.6) storage, respectively, and 5-, 4.5-, and 5.9-fold in CAPSO (pH 9.5) reaction buffers, respectively, compared with the wild-type enzyme (WT-$\small{D}$-PhgAT). In addition, all the fused $\small{D}$-PhgAT displayed higher enzymatic reaction rates than the WT-DPhgAT at all concentrations of L-glutamate monosodium salt used. The highest rate, $23.82{\pm}1.47$ mM/h, was that obtained from having ALAL-$\small{D}$-PhgAT reacted with 1,500 mM of the substrate. Moreover, the halophilic fusion significantly increased the tolerance of $\small{D}$-PhgAT in the presence of NaCl and KCl, being slightly in favor of KCl, where under the same condition at 3.5 M NaCl or KCl all halophilic-fused variants showed higher activity than WT-$\small{D}$-PhgAT.

Nutritional and Organoleptical Aspects of UHT Treated Milk (UHT처리된 우유의 영양 및 미각적 분석)

  • Hong, Youn-Ho
    • Korean Journal of Food Science and Technology
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    • v.14 no.3
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    • pp.276-282
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    • 1982
  • The UHT treatment and consequent storage effect on nutritional value of milk are discussed. Compared with the conventional sterilization the UHT treatment of milk represents a relatively small thermal stress. During UHT processing, nutritive value of protein, fat, carbohydrates, fat-soluble vitamins and minerals are generally unaffected. Nutritive value of some water soluble vitamins and protein are adversely affected in a small degree during storage. It has been recommended that UHT milk has best nutritional and organoleptic qualities on storage under refrigeration. Some unsolved future problems are also suggested.

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Effect of Feeding Yeast (Saceharomyces eerevisiae) on Growth Performance and Changes of Intestinal E. coli in Broiler Chicks (효모의 급여가 육계의 성장 및 장내 대장균의 변화에 미치는 영향)

  • 이현우;김인호;김춘수;손중천
    • Korean Journal of Poultry Science
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    • v.24 no.2
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    • pp.67-72
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    • 1997
  • In order to investigate the effect of feeding live yeast (Sacckaromyces cerevisiae) on the growth performance and changes of intestinal microorganism (E. coli), a growth assay was conducted with 144 broiler chicks. Treatments were consisted of corn-soybean meal control, 0.05% live yeast, and 0.05% dead yeast. Most of the chick protein of the live yeast was in the pure protein form, and had a high amino acid composition with 47% of essential amino acids and 53% of non-essential amino acids. No differences in growth performance were shown among dietany treatments. Total number of E. coli in the small intestine of chicks fed either live or dead yeast was significantly reduced compared to chicks fed the control diet. Although the changes of E. coli in the cecum were not identical to differences in the small intestine, the changes of E. coli in the cecurn had a similar trend.

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The Studies on the Interaction of Sulpyrin and Ampicillin (Sulpyrin과 Ampicillin의 상호작용(相互作用)에 관(關)한 연구(硏究))

  • Choi, Jun-Shik
    • Journal of Pharmaceutical Investigation
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    • v.10 no.3
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    • pp.20-26
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    • 1980
  • The purpose of this paper was to study the effect of sulpyrin on the absorption, excretion, metabolism, and protein binding of ampicillin in the small intestine of the rats and rabbits. The results are as follows; The absorption of ampicillin in small intestine of rats was increased by the combination of sulpyrin and ampicillin. The blood level of ampicillin in rabbits was elevated by oral administration of sulpyrin. The bioavailability of ampicillin was increased by simultaneous administration of sulpyrin and ampicillin. The urinary excretion of ampicillin was slightly decreased by combined administration of sulpyrin. The blood level of ampicillin was decreased and the urinary excretion was increased by long term administration of sulpyrin. On the other hand, metabolising enzyme of ampicillin was influenced by long term administration of sulpyrin. Protein binding rate of ampicillin was decreased by combination of sulpyrin as compared with control.

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Isolation and sequence analysis of a small cryptic plasmid from Lactobacillus farciminis KCTC3681 (Lactobacillus farciminis로부터 미지의 작은 플라스미드의 분리와 염기서열 분석)

  • Lee, Eun-Mo;Choi, Shin-Geon
    • Journal of Industrial Technology
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    • v.28 no.B
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    • pp.53-57
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    • 2008
  • From the extensive screening for small cryptic plasmid among about 23 lactic acid bacteria (LAB), 2.4 kb of cryptic plasmid was isolated from Lactobacillus farciminis strain KCTC 3681 and named as pLF24. The plasmid pLF24 was a circular molecule of 2,396 base-pairs in length with a G+C content of 38%. Two protein-coding sequences could be predicted. ORF1 and ORF2 showed homologies to plasmids of gram-positive bacteria. The replication protein coded by ORF2 and the plus origin, were similar to replication regions of other gram-positive bacteria as shown in plasmids such as pLH2, pLS141-1 and pLC2. The nucleotide sequence of pLF24 was deposited into Genbank data base with an accession number of EU429343. The newly isolated plasmid can be used for construction of shuttle vector in Lactobacillus bacteria.

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Chemical kinomics: a powerful strategy for target deconvolution

  • Kim, Do-Hee;Sim, Tae-Bo
    • BMB Reports
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    • v.43 no.11
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    • pp.711-719
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    • 2010
  • Kinomics is an emerging and promising approach for deciphering kinomes. Chemical kinomics is a discipline of chemical genomics that is also referred to as "chemogenomics", which is derived from chemistry and biology. Chemical kinomics has become a powerful approach to decipher complicated phosphorylation-based cellular signaling networks with the aid of small molecules that modulate kinase functions. Moreover, chemical kinomics has played a pivotal role in the field of kinase drug discovery as it enables identification of new molecular targets of small molecule kinase modulators and/or exploitation of novel functions of known kinases and has also provided novel chemical entities as hit/lead compounds. In this short review, contemporary chemical kinomics technologies such as activity-based protein profiling, T7 kinasetagged phages, kinobeads, three-hybrid systems, fluorescenttagged kinase binding assays, and chemical genomic profiling are discussed along with a novel allosteric Bcr-Abl kinase inhibitor (GNF-2/GNF-5) as a successful application of chemical kinomics approaches.