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RISS 인기검색어
- 검색키워드
- 저자 : Anandrao Ashok Patil (검색결과 5 건)
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Exosomes: Biogenesis, Composition, Functions, and Their Role in Pre-metastatic Niche Formation
Anandrao Ashok Patil,이원종 한국생물공학회 2019 Biotechnology and Bioprocess Engineering Vol.24 No.5
Exosomes are a subclass of extracellular vesicles that are released by normal or diseased cells. Exosomes are natural nanoparticles, contain natural cargo, including lipids, proteins, and nucleic acids, and are involved in intercellular communication. Exosomes are detected in the tumor microenvironment and participate in tumorigenesis by regulating angiogenesis, immunity, metastasis, and formation of pre-metastatic niches. This review mainly focuses on exosome biogenesis, composition, their biological and pathological functions, and roles in pre-metastatic niche formation.
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Cell death in culture: Molecular mechanisms, detections, and inhibition strategies
Anandrao Ashok Patil,Sachin Ashok Bhor,Won Jong Rhee 한국공업화학회 2020 Journal of Industrial and Engineering Chemistry Vol.91 No.-
Mammalian cell cultures are widely used in the biopharmaceutical industry to produce monoclonalantibodies, vaccines, growth factors, etc. Cell death is an essential biological process for physiologicalgrowth and development, but it is a major problem in biopharmaceutical production in bio-industry. Celldeath within bioreactor occurs due to various intracellular and extracellular stresses. These stressesnegatively affect the culture longevity, overall product quality, and yield. Among all cell death types,apoptosis accounts for most of the cellular death in the bioreactor. The implementation and developmentof various strategies to prevent the cellular death are crucial for robust bioprocess development. Celldeath during culture can be prevented or inhibited by supplementing media with specific chemicals,synthetic inhibitors, and genetic cell engineering approaches. In this review, we classified and describeddifferent types of cell death and their molecular mechanisms and summarized the cell death inhibitionapproaches implemented to inhibit cell death for various applications.
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Screening of lethal dsRNA and identification of dsRNA responsible genes in Tetranychus urticae
Deok Ho Kwon,Ji Hyun Park,Patil Anandrao Ashok,Unggyu Lee,Si Hyeock Lee 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.04
Tetranychus urticae is extremely hard to control by conventional acaricides due to its rapid development of resistance to nearly all arrays of acaricide. As an alternative control measure of acaricide-resistant mites, RNA interference (RNAi)-based method has recently been suggested. A double-stranded RNA (dsRNA) delivery method using multi-unit chambers was established and employed to screen the RNAi toxicity of 42 T. urticae genes. Among them, the dsRNA treatment of coatomer I (COPI) genes, such as coatomer subunit epsilon (COPE) and beta 2 (COPB2), resulted in high mortality [median lethal time (LT50) = 89.7 and 120.3 h, respectively]. The transcript level of the COPE gene was significantly (F3,9 = 16.2, P = 0.001) reduced up to 24% following dsRNA treatment, suggesting that the toxicity was likely mediated by the RNAi of the target gene. To identify the deferentially expressed gene upon dsRNA ingestion, RNA-seq was employed to compare the transcriptional profiles between mites fed dsEGFP and dsCOPB2. Approximately 928 of genes were up- or down-regulated significantly (P < 0.05) compared to control and 182 genes were commonly responded to the treatment of both dsRNAs. Those dsRNA-responsible genes were mainly categorized into metabolic enzymes, transporters and secretory proteins. Further study would be necessary to elucidate the roles of dsRNA-responsible genes in mite’s dsRNA uptake and defense.
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Deok Ho Kwon,Ji Hyun Park,Patil Anandrao Ashok,Unggyu Lee,Si Hyeock Lee 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10
Due to its rapid development of resistance to nearly all arrays of acaricide, Tetranychus urticae is extremely hard to control using conventional acaricides. As an alternative control measure of acaricide-resistant mites, RNA interference (RNAi)-based methods have recently been suggested. A double-stranded RNA (dsRNA) delivery method using multi-unit chambers was established and employed to screen the RNAi toxicity of 42 T. urticae genes. Among them, the dsRNA treatment of coatomer I (COPI) genes, such as coatomer subunit epsilon (COPE) and beta 2 (COPB2), resulted in high mortality [median lethal time (LT50) = 89.7 and 120.3h, respectively]. The transcript level of the COPE gene was significantly (F3,9 = 16.2, P =0.001) reduced by up to 24% following dsRNA treatment, suggesting that the toxicity was likely mediated by the RNAi of the target gene. As a toxicity enhancement strategy, the recombinant dsRNA was generated by reciprocally recombining half-divided fragments of COPE and COPB2. The two recombinant dsRNAs exhibited higher toxicity than the respective single dsRNA treatments as determined using LT50 values (79.2 and 81.5h, respectively). This finding indicates that the recombination of different genes can enhance RNAi toxicity and be utilized to generate synthetic dsRNA with improved RNAi efficacy.
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Jianping Chen,Jian Xu,Masato Hino,Mami Yamashita,Kazuma Hirata,Anandrao Ashok Patil,Tsuneyuki Tatsuke,Hiroaki Mon,Yutaka Banno,Takahiro Kusakabe,Jae Man Lee 한국응용곤충학회 2016 Journal of Asia-Pacific Entomology Vol.19 No.3
G protein-coupled receptors (GPCRs) are seven transmembrane proteins, which play an essential role in transmitting various extracellular signals into cells. For functional and structural analysis of GPCRs, it is necessary to produce active GPCRs in high quantitieswith outstanding purity. Fortunately, earlier baculovirus expression vector system has been reported as a proven functional GPCR mass-production tool. Therefore, in this study, we selected Bombyx mori allatostatin-C neuropeptide receptor BNGR-A1 as a GPCR reporter protein, which has been already proved successful binding of ligand. We confirmed its expression profile in various silkworm tissues and cell lines and then verified its plasma membrane subcellular localization in cultured silkworm BmN4 cells. In addition, we constructed recombinant baculoviruses for BNGR-A1, its ligand allatostatin-C (BmAST-C) and related eightG proteins (Gs, G12,Gα4, Gq, Gβ2,Gβ3, Gβ5 andGγ) and subsequentlymonitored the extracellular or intracellular expression of BNGR-A1 by co-infection with its ligand and cognate G proteins-expressing viruses. It is interesting to observe that different combinations of G proteins could result in changes or even undetectable of final yields of BNGR-A1, suggesting the essential roles of G proteins involved in the GPCR expression or stabilization. The present study demonstrated that co-infection of recombinant viruses expressing Gα4β3γ trimer enhanced the production of BNGR-A1 on BV fractions. To our knowledge, this is a fine strategy for identifying the specific G protein partner responsible for certain GPCR of interest. These studies would provide a novel idea for improving GPCR expression in the silkworm by baculovirus expression vector system.
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