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.

Thermal post-buckling analysis of uniform slender functionally graded material beams

Anandrao, K. Sanjay,Gupta, R.K.,Ramchandran, P.,Rao, G. Venkateswara Techno-Press 2010 Structural Engineering and Mechanics, An Int'l Jou Vol.36 No.5

Two or more distinct materials are combined into a single functionally graded material (FGM) where the microstructural composition and properties change gradually. Thermal post-buckling behavior of uniform slender FGM beams is investigated independently using the classical Rayleigh-Ritz (RR) formulation and the versatile Finite Element Analysis (FEA) formulation developed in this paper. The von-Karman strain-displacement relations are used to account for moderately large deflections of FGM beams. Bending-extension coupling arising due to heterogeneity of material through the thickness is included. Simply supported and clamped beams with axially immovable ends are considered in the present study. Post-buckling load versus deflection curves and buckled mode shapes obtained from both the RR and FEA formulations for different volume fraction exponents show an excellent agreement with the available literature results for simply supported ends. Response of the FGM beam with clamped ends is studied for the first time and the results from both the RR and FEA formulations show a very good agreement. Though the response of the FGM beam could have been studied more accurately by FEA formulation alone, the authors aim to apply the RR formulation is to find an approximate closed form post-buckling solutions for the FGM beams. Further, the use of the RR formulation clearly demonstrates the effect of bending-extension coupling on the post-buckling response of the FGM beams.

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.

Non-linear free vibrations and post-buckling analysis of shear flexible functionally graded beams

Anandrao, K. Sanjay,Gupta, R.K.,Ramchandran, P.,Rao, G. Venkateswara Techno-Press 2012 Structural Engineering and Mechanics, An Int'l Jou Vol.44 No.3

Large amplitude free vibration and thermal post-buckling of shear flexible Functionally Graded Material (FGM) beams is studied using finite element formulation based on first order Timoshenko beam theory. Classical boundary conditions are considered. The ends are assumed to be axially immovable. The von-Karman type strain-displacement relations are used to account for geometric non-linearity. For all the boundary conditions considered, hardening type of non-linearity is observed. For large amplitude vibration of FGM beams, a comprehensive study has been carried out with various lengths to height ratios, maximum lateral amplitude to radius of gyration ratios, volume fraction exponents and boundary conditions. It is observed that, for FGM beams, the non-linear frequencies are dependent on the sign of the vibration amplitudes. For thermal post-buckling of FGM beams, the effect of shear flexibility on the structural response is discussed in detail for different volume fraction exponents, length to height ratios and boundary conditions. The effect of shear flexibility is observed to be predominant for clamped beam as compared to simply supported beam.

Non-linear free vibrations and post-buckling analysis of shear flexible functionally graded beams

K. Sanjay Anandrao,R.K. Gupta,P. Ramchandran,G. Venkateswara Rao 국제구조공학회 2012 Structural Engineering and Mechanics, An Int'l Jou Vol.44 No.3

Large amplitude free vibration and thermal post-buckling of shear flexible Functionally Graded Material (FGM) beams is studied using finite element formulation based on first order Timoshenko beam theory. Classical boundary conditions are considered. The ends are assumed to be axially immovable. The von-Karman type strain-displacement relations are used to account for geometric non-linearity. For all the boundary conditions considered, hardening type of non-linearity is observed. For large amplitude vibration of FGM beams, a comprehensive study has been carried out with various lengths to height ratios, maximum lateral amplitude to radius of gyration ratios, volume fraction exponents and boundary conditions. It is observed that, for FGM beams, the non-linear frequencies are dependent on the sign of the vibration amplitudes. For thermal post-buckling of FGM beams, the effect of shear flexibility on the structural response is discussed in detail for different volume fraction exponents, length to height ratios and boundary conditions. The effect of shear flexibility is observed to be predominant for clamped beam as compared to simply supported beam.

Thermal post-buckling analysis of uniform slender functionally graded material beams

K. Sanjay Anandrao,R.K. Gupta,P. Ramchandran,G. Venkateswara Rao 국제구조공학회 2010 Structural Engineering and Mechanics, An Int'l Jou Vol.36 No.5

Two or more distinct materials are combined into a single functionally graded material (FGM) where the microstructural composition and properties change gradually. Thermal post-buckling behavior of uniform slender FGM beams is investigated independently using the classical Rayleigh-Ritz (RR) formulation and the versatile Finite Element Analysis (FEA) formulation developed in this paper. The von-Karman strain-displacement relations are used to account for moderately large deflections of FGM beams. Bending-extension coupling arising due to heterogeneity of material through the thickness is included. Simply supported and clamped beams with axially immovable ends are considered in the present study. Post-buckling load versus deflection curves and buckled mode shapes obtained from both the RR and FEA formulations for different volume fraction exponents show an excellent agreement with the available literature results for simply supported ends. Response of the FGM beam with clamped ends is studied for the first time and the results from both the RR and FEA formulations show a very good agreement. Though the response of the FGM beam could have been studied more accurately by FEA formulation alone, the authors aim to apply the RR formulation is to find an approximate closed form post-buckling solutions for the FGM beams. Further, the use of the RR formulation clearly demonstrates the effect of bending-extension coupling on the post-buckling response of the FGM beams.

Screening of lethal genes for RNAi and toxicity enhancement by recombination in Tetranychus urticae Koch

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.

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.

Development and evaluation of psyllium seed husk polysaccharide based wound dressing films

Basavaraj S. Patil,Vinayak S. Mastiholimath,Anandrao R. Kulkarni 경희대학교 융합한의과학연구소 2011 Oriental Pharmacy and Experimental Medicine Vol.11 No.2

In the present study, wound dressing films were fabricated using Seed Husk of Psyllium (SHP) complexed with povidone iodine and were evaluated for various physicochemical properties as well as wound healing activity in albino rats. The required film properties for the successful wound dressings, such as film elongation (elasticity), tensile strength, water uptake and water vapor transmission rates (WVTR) were examined. Results indicated that SHP films showed good elasticity (80.27–116.24%) and acceptable tensile strengths (8.33–22.13 N/mm2). SHP films were evaluated for water absorption pattern as well as permeation to water vapours and results showed that films were able to swell in water upto 167 to 191% w/w and WVTR were in the range between 8.23×10−3 to 1.12×10−4 g.cm2/day. Selected formulation of SHP films with optimum characteristics were loaded with Povidone iodine (PI) solution by soaking method and were evaluated for antimicrobial and wound healing activity on excision wound model. Results indicated that PI loaded films showed significant antibacterial activity against both gram positive and gram negative selected bacteria. PI loaded SHP films showed comparatively better wound healing property to control group with faster epithelialization and greater rates of wound contraction.

Characterization of modular bifunctional processive endoglucanase Cel5 from Hahella chejuensis KCTC 2396

Ghatge, Sunil Subhash,Telke, Amar Anandrao,Kang, Seo-Hee,Arulalapperumal, Venkatesh,Lee, Keun-Woo,Govindwar, Sanjay Prabhu,Um, Youngsoon,Oh, Doo-Byoung,Shin, Hyun-Dong,Kim, Seon-Won Springer-Verlag 2014 Applied microbiology and biotechnology Vol.98 No.10