Search results for: heat shock protein expression

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: A.Kouidri, B.Madani

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The present work deals with the study of the heat transfer in a rectangular channel equipped with a metallic foam. The tested metallic foam sample is made from copper with 20 PPI (Pore per Inch Linear) and 93% of porosity and the working fluid used is the n-pentane. In the present work the independent variables are the velocity in the range from 0.02 to 0.06 m/s and a boiling heat flux rate varying between 30 and 70 kW/m2. The heat transfer coefficient is presented versus boiling heat flux, vapor quality and superheat ΔTsat. The thermal results are compared to those found for a plain tube for the same conditions. The comparison with the plain tube shows that the insert of a metallic foam enhances the heat transfer coefficient by a factor between 1.3 and 3.

Keywords: boiling, metallic foam, heat transfer, plate heat exchanger

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Authors: Kathy Stein, Mariola Lysson, Anja Schmidt, Beatrix Schumak, Sabine Specht, Hicham Bouabe, Jürgen Heesemann, Axel Roers, Joerg C. Kalff, Sven Wehner

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Objective: Postoperative ileus (POI) is a common complication of abdominal surgery. Monocyte infiltration is a hallmark of POI. The polarization of macrophages/monocytes in this process is not well understood. We aimed to investigate if and how M2 macrophage/monocyte differentiation is involved in POI pathogenesis. Design: POI was induced by intestinal manipulation (IM). C57Bl/6, CCR2-/-, IL-10 reporter (ITIB), IL-10-/- and LysMcre/IL-10fl/fl mice underwent IM. At various points in time leukocyte influx, gene and protein expression of cytokines, chemokines and M2 differentiation markers and intestinal motility were analyzed. Results: IM induced the postoperative expression of the M2 markers Arginase-1 and YM-1, predominantly in F4/80+Ly6C+ monocytes. Gene expression analyses indicated an IL-10-dependent, IL-4-independent M2 polarization of these monocytes. IL-10 dependency of M2 differentiation was confirmed in IL-10 deficient mice. Leukocytes, in the order of infiltrating monocytes, neutrophils, and resident macrophages were the main IL-10 producers during POI. IL-10 producing monocytes as well as M2 marker expression were almost absent in CCR2-deficient mice. However, postoperative IL-10 expression was not altered in CCR2-/- mice. The loss of M2 polarized monocytes neither protected CCR2-/- mice from nor affected resolution of POI. In contrast, IL-10 deficiency reduced postoperative neutrophil numbers and ameliorated POI. IL-10Ra expression was strongly induced in neutrophils but not in monocytes. Conclusion: We conclude that IL-10 counteracts POI resolution by activating IL-10Ra-expressing neutrophils in the late phase of disease while IL-10-dependent M2 differentiation is not pivotal to POI manifestation and resolution.

Keywords: interleukin-10, macrophages, neutrophils, postoperative ileus

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Authors: H. Shokouhmand, A. Ghanami

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Heat pipe is simple heat transfer device which combines the conduction and phase change phenomena to control the heat transfer without any need for external power source. At hot surface of heat pipe, the liquid phase absorbs heat and changes to vapor phase. The vapor phase flows to condenser region and with the loss of heat changes to liquid phase. Due to gravitational force the liquid phase flows to evaporator section. In HVAC systems the working fluid is chosen based on the operating temperature. The heat pipe has significant capability to reduce the humidity in HVAC systems. Each HVAC system which uses heater, humidifier or dryer is a suitable nominate for the utilization of heat pipes. Generally heat pipes have three main sections: condenser, adiabatic region and evaporator.Performance investigation and optimization of heat pipes operation in order to increase their efficiency is crucial. In present article, a parametric study is performed to improve the heat pipe performance. Therefore, the heat capacity of heat pipe with respect to geometrical and confining parameters is investigated. For the better observation of heat pipe operation in HVAC systems, a CFD simulation in Eulerian- Eulerian multiphase approach is also performed. The results show that heat pipe heat transfer capacity is higher for water as working fluid with the operating temperature of 340 K. It is also showed that the vertical orientation of heat pipe enhances it’s heat transfer capacity used in the abstract.

Keywords: heat pipe, HVAC system, grooved heat pipe, CFD simulation

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Authors: M. Kouyate, M. Sangare, S. Samake, S. Keita, H. G. Kim, D. H. Geschwind

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Background & Objectives: Human genetic studies can be expensive, even unaffordable, in developing countries, partly due to the sequencing costs. Our aim is to pilot the use of bioinformatics tools to guide scientifically valid, locally relevant, and economically sound autism genetic research in Mali. Methods: The following databases, NCBI, HGMD, and LSDB, were used to identify hot point mutations. Phenotype, transmission pattern, theoretical protein expression in the brain, the impact of the mutation on the 3D structure of the protein) were used to prioritize selected autism genes. We used the protein database, Modeller, and clustal W. Results: We found Mef2c (Gly27Ala/Leu38Gln), Pten (Thr131IIle), Prodh (Leu289Met), Nme1 (Ser120Gly), and Dhcr7 (Pro227Thr/Glu224Lys). These mutations were associated with endonucleases BseRI, NspI, PfrJS2IV, BspGI, BsaBI, and SpoDI, respectively. Gly27Ala/Leu38Gln mutations impacted the 3D structure of the Mef2c protein. Mef2c protein sequences across species showed a high percentage of similarity with a highly conserved MADS domain. Discussion: Mef2c, Pten, Prodh, Nme1, and Dhcr 7 gene mutation frequencies in the Malian population will be very informative. PCR coupled with restriction enzyme digestion can be used to screen the targeted gene mutations. Sanger sequencing will be used for confirmation only. This will cut down considerably the sequencing cost for gene-to-gene mutation screening. The knowledge of the 3D structure and potential impact of the mutations on Mef2c protein informed the protein family and altered function (ex. Leu38Gln). Conclusion & Future Work: Bio-informatics will positively impact autism research in Mali. Our approach can be applied to another neuropsychiatric disorder.

Keywords: bioinformatics, endonucleases, autism, Sanger sequencing, point mutations

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Authors: Mohamed Abdullah Ahmed

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In a study of chickpea protein isolate (CPI) preparation, the wet alkaline extraction was carried out. The objectives were to determine the optimal extracting conditions of CPI and apply CPI into a sponge cake recipe to replace egg and make acceptable product. The design used in extraction was a central composite design. The response surface methodology was preferred to graphically express the relationship between extraction time and pH with the output variables of percent yield and protein content of CPI. It was noted that optimal extracting conditions were 60 min and pH 10.5 resulting in 90.07% protein content and 89.15% yield of CPI. The protein isolate (CPI) could be incorporated in cake to 20% without adversely affecting the cake physical properties such as cake hardness and sensory attributes. The higher protein content in cake was corresponding to the amount of CPI added. Therefore, adding CPI can significantly (p<0.05) increase protein content in cake. However, sensory evaluation showed that adding more than 20% of CPI decreased the overall acceptability. The results of this investigation could be used as a basic knowledge of CPI utilization in other food products.

Keywords: chick bean protein isolate, sponge cake, utilization, sponge

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Authors: Alhadi Bustaman, Soeganda Formalidin, Titin Siswantining

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DNA microarray technology is used to analyze thousand gene expression data simultaneously and a very important task for drug development and test, function annotation, and cancer diagnosis. Various clustering methods have been used for analyzing gene expression data. However, when analyzing very large and heterogeneous collections of gene expression data, conventional clustering methods often cannot produce a satisfactory solution. Biclustering algorithm has been used as an alternative approach to identifying structures from gene expression data. In this paper, we introduce a transform technique based on singular value decomposition to identify normalized matrix of gene expression data followed by Mixed-Clustering algorithm and the Lift algorithm, inspired in the node-deletion and node-addition phases proposed by Cheng and Church based on Agglomerative Hierarchical Clustering (AHC). Experimental study on standard datasets demonstrated the effectiveness of the algorithm in gene expression data.

Keywords: agglomerative hierarchical clustering (AHC), biclustering, gene expression data, lymphoma, singular value decomposition (SVD)

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Authors: Hugo Reis, Isabel Rabiais

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The main purpose of the present study is to understand the nurse’s responsibility in preventing the septic shock in an oncological patient with febrile neutropenia submitted to chemotherapy. In order to do it, an integrative review of literature has been conducted. In the research done in many databases, it was concluded that only 7 out of 5202 articles compiled the entire inclusion standard present in the strict protocol of research, being this made up by all different methodologies. On the research done in the 7 articles it has resulted 8 text macro-units associated to different nursing interventions: ‘Health Education’; ‘Prophylactic Therapy Administration’; ‘Scales Utilization’; ‘Patient Evaluation’; ‘Environment Control’; ‘Performance of Diagnostic Auxiliary Exams’; ‘Protocol Enforcement/Procedure Guidelines’; ‘Antibiotic Therapy Administration’. Concerning the prevalence/result’s division there can be identified many conclusions: the macro-units ‘Patient Evaluation’, ‘Performance of Diagnostic Auxiliary Exams’, and ‘Antibiotic Therapy Administration’ present themselves to be the most prevalent in the research – 6 in 7 occurrences (approximately 85.7%). Next, the macro-unit ‘Protocol Enforcement/Procedure Guidelines’ presents itself as an important expression unit – being part of 5 out of the 7 analyzed studies (approximately 71.4%). The macro-unit ‘Health Education’, seems to be in the same way, an important expression unit – 4 out of the 7 (or approximately 57%). The macro-unit ‘Scales Utilization’, represents a minor part in the research done – it’s in only 2 out of the 7 cases (approximately 28.6%). On the other hand, the macro-units ‘Prophylactic Therapy Administration’ and ‘Environment Control’ are the two categories with fewer results in the research - 1 out of the 7 cases, the same as approximately 14.3% of the research results. Every research done to the macro-unit ‘Antibiotic Therapy Administration’ agreed to refer that the intervention should be strictly done, in a period of time less than one hour after diagnosing the fever, with the purpose of controlling the quick spread of infection – minimizing its seriousness. Identifying these interventions contributes, concluding that, to adopt strategies in order to prevent the phenomenon that represents a daily scenario responsible for the cost´s increase in health institutions, being at the same time responsible for the high morbidity rates and mortality increase associated with this specific group of patients.

Keywords: febrile neutropenia, oncology nursing, patient, septic shock

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Authors: Thanusha D. Abeywickrama, Inoka C. Perera, Genji Kurisu

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Tuberculosis, considered being as the ninth leading cause of death worldwide, cause from a single infectious agent M. tuberculosis and the drug resistance nature of this bacterium is a continuing threat to the world. Therefore TB preventing treatment is expanding, where this study designed to analyze the regulatory mechanism of GntR transcriptional regulator gene Rv0792c, which lie between several genes codes for some hypothetical proteins, a monooxygenase and an oxidoreductase. The gene encoding Rv0792c was cloned into pET28a and expressed protein was purified to near homogeneity by Nickel affinity chromatography. It was previously reported that the protein binds within the intergenic region (BS region) between Rv0792c gene and monooxygenase (Rv0793). This resulted in binding of three protein molecules with the BS region suggesting tight control of monooxygenase as well as its own gene. Since monooxygenase plays a key role in metabolism, this gene may have a global regulatory role. The natural ligand for this regulator is still under investigation. In relation to the Rv0792 protein structure, a Circular Dichroism (CD) spectrum was carried out to determine its secondary structure elements. Percentage-wise, 17.4% Helix, 21.8% Antiparallel, 5.1% Parallel, 12.3% turn and 43.5% other were revealed from CD spectrum data under room temperature. Differential Scanning Calorimetry (DSC) was conducted to assess the thermal stability of Rv0792, which the melting temperature of protein is 57.2 ± 0.6 °C. The graph of heat capacity (Cp) versus temperature for the best fit was obtained for non-two-state model, which concludes the folding of Rv0792 protein occurs through stable intermediates. Peak area (∆HCal ) and Peak shape (∆HVant ) was calculated from the graph and ∆HCal / ∆HVant was close to 0.5, suggesting dimeric nature of the protein.

Keywords: CD spectrum, DSC analysis, GntR transcriptional regulator, protein structure

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Authors: Derya Yucel, Nigar Angın, Dürdane Mart, Meltem Turkeri, Volkan Catalkaya, Celal Yucel

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Chickpea (Cicer arietinum L.) with high protein content is a vital food, especially in under-developed and developing countries for the people who do not consume enough meat due to low-income level. The objective of the proposed study is to evaluate growing, yield and yield components of chickpea genotypes under Mediterranean condition so determine tolerance of chickpea genotypes against drought and heat stress. For this purpose, a total of 34 chickpea genotypes were used as material. The experiment was conducted according to factorial randomized complete block design with 3 reps at the Eastern Mediterranean Research Institute, Adana, TURKEY for 2014-15 growing season under three different growing conditions (Winter sowing, irrigated-late sowing and non-irrigated- late sowing). According to results of this experiment, vegetative period, flowering time, poding time, maturity time, plant height, height of first pod, seed yield and 100 seed weight were ranged between 68.33 to 78.77 days, 94.22 to 85.00 days, 94.11 to 106.44 days, 198.56 to 214.44 days, 37.18 to 64.89 cm, 18.33 to 34.83 cm, 417.1 to 1746.4 kg/ha and 14.02 to 45.02 g, respectively. Among the chickpea genotypes, the Aksu, Arda, Çakır, F4 09 (X 05 TH 21-16189), FLIP 03-108 were least affected by drought and heat stress. Therefore, these genotypes can be used as sources of drought and heat tolerance in further breeding programme for evolving the drought and heat tolerant genotypes in chickpea.

Keywords: chickpea, drought stress, heat stress, yield

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Authors: Cheng-Cao Sun, Shu-Jun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, De-Jia Li

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Hsa-miRNA-326 (miR-326) has recently been discovered having anticancer efficacy in different organs. However, the role of miR-326 on non-small cell lung cancer (NSCLC) is still ambiguous. In this study, we investigated the role of miR-326 on the development of NSCLC. The results indicated that miR-326 was significantly down-regulated in primary tumor tissues and very low levels were found in NSCLC cell lines. Ectopic expression of miR-326 in NSCLC cell lines significantly suppressed cell growth as evidenced by cell viability assay, colony formation assay and BrdU staining, through inhibition of cyclin D1, cyclin D2, CDK4, and up-regulation of p57(Kip2) and p21(Waf1/Cip1). In addition, miR-326 induced apoptosis, as indicated by concomitantly with up-regulation of key apoptosis protein cleaved caspase-3, and down-regulation of anti-apoptosis protein Bcl2. Moreover, miR-326 inhibited cellular migration and invasiveness through inhibition of matrix metalloproteinases (MMP)-7 and MMP-9. Further, oncogene CCND1 was revealed to be a putative target of miR-326, which was inversely correlated with miR-326 expression in NSCLC. Taken together, our results demonstrated that miR-326 played a pivotal role on NSCLC through inhibiting cell proliferation, migration, invasion, and promoting apoptosis by targeting oncogenic CCND1.

Keywords: hsa-miRNA-326 (miR-326), cyclin D1, non-small cell lung cancer (NSCLC), proliferation, apoptosis

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Authors: M. Madheswaran, A. R. Prashant, S. Ramakrishna, V. Ramesh Naidu, P. Govindan, P. Aravindakshan

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Shock Response spectrum (SRS) tests are one of the tests that are conducted on some critical systems of spacecraft as part of environmental testing. The SRS tests are conducted to simulate the pyro shocks that occur during launch phases as well as during deployment of spacecraft appendages. Some of the methods to carryout SRS tests are pyro technique method, impact hammer method, drop shock method and using electro dynamic shakers. The pyro technique, impact hammer and drop shock methods are open loop tests, whereas SRS testing using electrodynamic shaker is a controlled closed loop test. SRS testing using electrodynamic shaker offers various advantages such as simple test set up, better controllability and repeatability. However, it is important to devise a a proper test methodology so that safety of the electro dynamic shaker and that of test specimen are not compromised. This paper discusses the challenges that are involved in conducting SRS tests, shaker validation and the necessary precautions to be considered. Approach involved in choosing various test parameters like synthesis waveform, spectrum convergence level, etc., are discussed. A case study of SRS test conducted on an optical payload of Indian Geo stationary spacecraft is presented.

Keywords: maxi-max spectrum, SRS (shock response spectrum), SDOf (single degree of freedom), wavelet synthesis

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Authors: Ali Bayram, Remzi Yiğiter

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Objective: Alzheimer's disease (AD), the most common cause of dementia, is a progressive neurodegenerative disease. At present, diagnosis of AD is rather late in the disease. Therefore, we attempted to find peripheral biomarkers for the early diagnosis of AD. Herein, we conducted a study to investigate the unc-51 like autophagy activating kinase-1 (ULK1) mRNA expression levels in human peripheral blood mononuclear cells from patients with Alzheimer's disease. Method: To determine whether ULK1 gene expression are altered in AD patients, we measured their gene expression in human peripheral blood cell in 50 patients with AD and 50 age and gender matched healthy controls by quantitative real-time PCR technique. Results: We found that both ULK1 gene expression in peripheral blood cell were significantly decreased in patients with AD as compared with controls (p <0.05). Lower levels of ULK1 gene expression were significantly associated with the increased risk for AD. Conclusions: Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2, and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. Alzheimer is the most common neurodegenerative disease. Our results provide useful information that the ULK1 gene expression is decreased in the neurodegeneration and AD patients with, indicating their possible systemic involvement in AD.

Keywords: Alzheimer’s sisease, ULK1, mRNA expression, RT-PCR

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Authors: Mihai Palade, Gina Cecilia Pistol, Mariana Stancu, Veronica Chedea, Ionelia Taranu

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Nutrition is an important determinant of general health status, with especially focus on prevention and/or attenuation of the inflammatory-associated pathologies. People with chronic kidney disease can experience chronic inflammation that can lead to cardiovascular disease and even an increased rate of death. There are important links between chronic kidney diseases, inflammation and nutritional strategies that may prevent or protect against undesirable inflammation and oxidative stress. The grape by-products either seeds or pomace are rich in polyphenols which may be beneficial in prevention of inflammatory, antioxidant and antimicrobial processes. As a model for studying the impact of grape seeds on renal inflammation and oxidative stress, we used in this study weaned piglets. After a feeding trial of 30 days with a control diet and an experimental diet containing 5% grape seed (GS), kidney samples were collected. In renal tissues were determined the expression and activity of important markers of immune respose and oxidative stress: pro-inflammatory cytokines (TNF-alpha, IL-1 beta, IL-6, IL-8, IFN-gamma), anti-inflammatory cytokines (IL-4, IL-10), anti-oxidant enzymes (catalase CAT, superoxide dismutase SOD, glutathione peroxidise GPx) and important mediators belonging to nuclear receptors (NF-kB1, Nrf-2 and PPAR-gamma). Gene expression was evaluated by qPCR, whereas protein concentration was determined using proteomic techniques (ELISA). The activity of anti-oxidant enzymes was determined using specific kits. Our results showed that GS enriched in polyphenols does not have effect on TNF-alpha, IL-6 and IL-1 beta gene expression and protein concentration in kidney. By contrast, the gene expression and protein level of IL-8 and IFN-gamma were decreased in GS kidney. Anti-inflammatory cytokines IL-4 and IL-10 gene levels were increased in kidneys collected from GS piglets in comparison with controls, with no modification of protein levels between the two groups. The activities of anti-oxidant enzymes CAT and GPx were increased in kidney by GS, whereas SOD activity was unmodified in comparison with control samples. Also, the GS diet was associated with no modulation of mRNAs for nuclear receptors NF-kB1, Nrf-2 and PPAR-gamma gene expressions in kidneys. In conclusion, our results demonstrated that GS enriched in bioactive compounds such polyphenols could modulate inflammation and oxidative stress markers in kidney tissues. Further studies are necessary to elucidate the mechanism of action of GS compounds in case kidney inflammation associated with oxidative stress, and signalling molecules involved in these mechanisms.

Keywords: animal model, kidney inflammation, oxidative stress, grape seed

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Authors: Akterono Budiyati, Gita Kusumo, Teguh Putra, Fritzie Rexana, Antonius Kurniawan, Aru Sudoyo, Ahmad Utomo, Andi Utama

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The presence of the programmed death ligand-1 (PD-L1) has been used in multiple clinical trials and approved as biomarker for selecting patients more likely to respond to immune checkpoint inhibitors. However, the expression of PD-L1 is regulated in different ways, which leads to a different significance of its presence. Positive PD-L1 within tumors may result from two mechanisms, induced PD-L1 expression by T-cell presence or genetic mechanism that lead to constitutive PD-L1 expression. Amplification of PD-L1 genes was found as one of genetic mechanism which causes an increase in PD-L1 expression. In case of colorectal cancer (CRC), targeting immune checkpoint inhibitor has been recommended for patients with microsatellite instable (MSI). Although the correlation between PD-L1 expression and MSI status has been widely studied, so far the precise mechanism of PD-L1 gene activation in CRC patients, particularly in MSI population have yet to be clarified. In this present study we have profiled 61 archived formalin fixed paraffin embedded CRC specimens of patients from Medistra Hospital, Jakarta admitted in 2010 - 2016. Immunohistochemistry was performed to measure expression of PD-L1 in tumor cells as well as MSI status using antibodies against PD-L1 and MMR (MLH1, MSH2, PMS2 and MSH6), respectively. PD-L1 expression was measured on tumor cells with cut off of 1% whereas loss of nuclear MMR protein expressions in tumor cells but not in normal or stromal cells indicated presence of MSI. Subset of PD-L1 positive patients was then assessed for copy number variations (CNVs) using single Tube TaqMan Copy Number Assays Gene CD247PD-L1. We also observed KRAS mutation to profile possible genetic mechanism leading to the presence or absence of PD-L1 expression. Analysis of 61 CRC patients revealed 15 patients (24%) expressed PD-L1 on their tumor cell membranes. The prevalence of surface membrane PD-L1 was significantly higher in patients with MSI (87%; 7/8) compared to patients with microsatellite stable (MSS) (15%; 8/53) (P=0.001). Although amplification of PD-L1 gene was not found among PD-L1 positive patients, low-level amplification of PD-L1 gene was commonly observed in MSS patients (75%; 6/8) than in MSI patients (43%; 3/7). Additionally, we found 26% of CRC patients harbored KRAS mutations (16/61), so far the distribution of KRAS status did not correlate with PD-L1 expression. Our data suggest genetic mechanism through amplification of PD-L1 seems not to be the mechanism underlying upregulation of PD-L1 expression in CRC patients. However, further studies are warranted to confirm the results.

Keywords: colorectal cancer, gene amplification, microsatellite instable, programmed death ligand-1

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Authors: Anupoju Rajeev, Joanne Mathew, Amit Shelke

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In the current study, a new type of Composite Structural Insulated Panels (CSIPs) is developed and investigated its performance against shock loading which can replace the conventional wooden structural materials. The CSIPs is made of Fibre Cement Board (FCB)/aluminum as the facesheet and the expanded polystyrene foam as the core material. As tornadoes are very often in the western countries, it is suggestable to monitor the health of the CSIPs during its lifetime. So, the composite structure is installed with three smart sensors located randomly at definite locations. Each smart sensor is fabricated with an embedded half stainless phononic crystal sensor attached to both ends of the nylon shaft that can resist the shock and impact on facesheet as well as polystyrene foam core and safeguards the system. In addition to the granular crystal sensors, the accelerometers are used in the horizontal spanning and vertical spanning with a definite offset distance. To estimate the health and damage of the CSIP panel using granular crystal sensor, shock wave loading experiments are conducted. During the experiments, the time of flight response from the granular sensors is measured. The main objective of conducting shock wave loading experiments on the CSIP panels is to study the effect and the sustaining capacity of the CSIP panels in the extreme hazardous situations like tornados and hurricanes which are very common in western countries. The effects have been replicated using a shock tube, an instrument that can be used to create the same wind and pressure intensity of tornado for the experimental study. Numerous experiments have been conducted to investigate the flexural strength of the CSIP. Furthermore, the study includes the damage detection using three smart sensors embedded in the CSIPs during the shock wave loading.

Keywords: composite structural insulated panels, damage detection, flexural strength, sandwich structures, shock wave loading

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Authors: Stefan Gruden, Charlott Brunmark, Bo Holmqvist, Erwin D. Brenndorfer, Martin Johansson, Jian Liu, Ying Zhao, Niklas Axen, Moustapha Hassan

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Aim: The study was designed to evaluate the ability of the calcium sulfate-based NanoZolid® drug delivery technology to locally release the epidermal growth factor (EGF) protein while maintaining its biological activity. Methods: NanoZolid-formulated EGF protein labelled with a near-infrared dye (EGF-NIR) depots or EGF-NIR dissolved in PBS were injected subcutaneously into mice bearing EGF receptor (EGFR) positive human A549 lung cancer tumors inoculated subcutaneously. The release and biodistribution of the EGF-NIR were investigated in vivo longitudinally up to 96 hours post-administration, utilizing whole-body fluorescence imaging. In order to confirm the in vivo findings, histological analysis of tumor cryosections was performed to investigate EGF-NIR fluorescent signal and EGFR expression level by immunofluorescence labelling. Results: The in vivo fluorescence imaging showed a controlled release profile of the EGF-NIR loaded in the NanoZolid depots compared to free EGF-NIR. Histological analysis of the tumors further demonstrated a prevailing distribution of EGF-NIR in regions with high levels of EGFR expression. Conclusion: Calcium sulfate based depots can be used to formulate EGF while maintaining its biological activity, e.g., receptor binding capability. This may have good clinical potential for local delivery of biomolecules to enhance treatment efficacy and minimize systemic adverse effects.

Keywords: bioresorbable, calcium sulfate, controlled release, NanoZolid

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Authors: Muhammad Shahrukh Saeed, Syed Ahmad Nameer, Shafiq Ur Rehman, Aisha Jillani

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Heat exchangers play a critical role in industrial applications of thermal systems. Its physical size and performance are vital parameters; therefore enhancement of heat transfer through different techniques remained a major research area for both academia and industry. This research reports the main purpose of heat exchanger with better kit design which plays a vital role during the process of heat transfer. Plate type heat exchanger mainly requires a design in which the plates can be easily be installed and removed without having any problem with the plates. For the flow of the fluid within the heat exchanger, it requires a flow should be fully developed. As natural laws allows the driving energy of the system to flow until equilibrium is achieved. As with a plate type heat exchanger heat the heat penetrates the surface which separates the hot medium with the cold one very easily. As some of the precautions should be considered while taking the heat exchanger accountable like heat should transfer from hot medium to cold, there should always be difference in temperature present and heat loss from hot body should be equal to the heat gained by the cold body regardless of the losses present to the surroundings. Aluminum plates of same grade are used in all experiments to ensure similarity. Size of all plates was 254 mm X 100 mm and thickness was taken as 5 mm.

Keywords: heat transfer coefficient, aluminium, entry length, design

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Authors: Shail N. Shah, K. K. Baraya, A. Madhusudan Achari

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Loop Heat Pipe is a passive two-phase heat transfer device which can be used without any external power source to transfer heat from source to sink. The main aim of this paper is to have modelling of water-based LHP at varying heat loads. Through figures, how the fluid flow occurs within the loop has been explained. Energy Balance has been done in each section. IC (Iterative Convergence) scheme to find out the SSOT (Steady State Operating Temperature) has been developed. It is developed using Dev C++. To best of the author’s knowledge, hardly any detail is available in the open literature about how temperature distribution along the loop is to be evaluated. Results for water-based loop heat pipe is obtained and compared with open literature and error is found within 4%. Parametric study has been done to see the effect of different parameters on pressure drop and SSOT at varying heat loads.

Keywords: loop heat pipe, modelling of loop heat pipe, parametric study of loop heat pipe, functioning of loop heat pipe

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Authors: Sachin K. Samuchiwal, Barbara Balestrieri, Amanda Paskavitz, Hannah Raff, Joshua A. Boyce

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IL-33, a recently discovered member of the IL-1 cytokine family, binds to the TLR/IL1R super family receptor ST2 and induces type 2 immune responses. IL-33 is constitutively expressed in structural cells at barrier sites such as skin, lung, and intestine, and also inducibly expressed by hematopoietic cells including macrophages. Stimulation of macrophages by Lipopolysaccharide (LPS) can induce de novo IL-33 expression, and also causes the production of prostaglandin-E2 (PGE2) via cyclooxygenase (COX)-2 and microsomal PGE2 synthase-1 (mPGES-1). Because PGE2 can regulate macrophage functions through both autocrine and paracrine mechanisms, the potential interplay of endogenous PGE2 on IL-33 production was explored. Bone-marrow derived murine macrophages (bmMF) that lack either mPGES-1 or EP2 receptor expression were stimulated with LPS in the absence or presence of exogenous PGE2 along with pharmacological agonists and antagonists. The study results demonstrate that endogenous PGE2 markedly enhances LPS-induced IL-33 production by bmMFs via EP2 receptors. Moreover, exogenous PGE2 can amplify LPS-induced IL-33 expression dominantly by EP2 and partly by EP4 receptors by a pathway involving cAMP and exchange protein activated by cAMP (EPAC), but not protein kinase A (PKA). Though both IL-33 production and PGE2 generation in response to LPS require activation of both p38 MAPK and NF-κB, PGE2 did not influence this activation. In conclusion, it is demonstrated that endogenous PGE2 signaling through EP2 and EP4 receptors is a prerequisite for LPS-induced IL-33 production in bmMFs and the underlying cAMP mediated pathway involves EPAC. Since IL-33 is a critical pro-inflammatory cytokine in various pathological disorders, this PGE2-EP2/EP4-cAMP mediated pathway can be exploited to intervene in IL-33 driven pathologies.

Keywords: bone marrow macrophages, EPAC, IL-33, PGE2

Procedia PDF Downloads 161

Authors: Filip Franciszek Karuga, Szymon Turkiewicz, Marta Ditmer, Marcin Sochal, Piotr Białasiewicz, Agata Gabryelska

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Circadian rhythm, an internal coordinator of physiological processes is composed of a set of semi-autonomous clocks. Clocks are regulated through the expression of circadian clock genes which form feedback loops, creating an oscillator. The primary loop consists of activators: CLOCK, BMAL1 and repressors: CRY, PER. CLOCK can be substituted by the Neuronal PAS Domain Protein 2 (NPAS2). Orphan nuclear receptor (REV-ERB-α) is a component of the secondary major loop, modulating the expression of BMAL1. Circadian clocks might be disrupted by the obstructive sleep apnea (OSA), which has also been associated with type II diabetes mellitus (DM2). Interestingly, studies suggest that dysregulation of NPAS2 and REV-ERB-α might contribute to the pathophysiology of DM2 as well. The goal of our study was to examine the role of NPAS2 and REV-ERB-α in DM2 in OSA patients. After examination of the clinical data, all participants underwent polysomnography (PSG) to assess their apnea-hypopnea index (AHI). Based on the acquired data participants were assigned to one of 3 groups: OSA (AHI>30, no DM2; n=17 for NPAS2 and 34 for REV-ERB-α), DM2 (AHI>30 + DM2; n=7 for NPAS2 and 15 for REV-ERB-α) and control group (AHI<5, no DM2; n=16 for NPAS2 and 31 for REV-ERB-α). ELISA immunoassay was performed to assess the serum protein level of REV-ERB-α and NPAS2. The only statistically significant difference between groups was observed in NPAS2 protein level (p=0.037). Post-hoc analysis showed significant differences between the OSA and the control group (p=0.017). AHI and NPAS2 level was significantly correlated (r=-0.478, p=0.002) in all groups. A significant correlation was observed between the REV-ERB-α level and sleep efficiency (r=0.617, p=0.005) as well as sleep maintenance efficiency (r=0.645, p=0.003) in the OSA group. We conclude, that NPAS2 is associated with OSA severity and might contribute to metabolic sequelae of this disease. REV-ERB-α on the other hand can influence sleep continuity and efficiency.

Keywords: OSA, diabetes mellitus, endocrinology, chronobiology

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Authors: Kemal Comakli, Meryem Terhan

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In this study, the potential of heat recovery from waste flue gas was examined in 60 MW district heating system of a university, and fuel saving was aimed by using the recovered heat in the system as a source again. Various scenarios are intended to make use of waste heat. For this purpose, actual operation data of the system were taken. Besides, the heat recovery units that consist of heat exchangers such as flue gas condensers, economizers or air pre-heaters were designed theoretically for each scenario. Energy analysis of natural gas-fired boiler’s exhaust flue gas in the system, and economic analysis of heat recovery units to predict payback periods were done. According to calculation results, the waste heat loss ratio from boiler flue gas in the system was obtained as average 16%. Thanks to the heat recovery units, thermal efficiency of the system can be increased, and fuel saving can be provided. At the same time, a huge amount of green gas emission can be decreased by installing the heat recovery units.

Keywords: heat recovery from flue gas, energy analysis of flue gas, economical analysis, payback period

Procedia PDF Downloads 265

Authors: Laszlo Vari

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In the digital age, the spread of the mobile world and the nature of the cyberspace, offers many new opportunities for the prevalence of the fundamental right to free expression, and therefore, for free speech and freedom of the press; however, these new information communication technologies carry many new challenges. Defamation, censorship, fake news, misleading information, hate speech, breach of copyright etc., are only some of the violations, all of which can be derived from the harmful exercise of freedom of expression, all which become more salient in the internet. Here raises the question: how can we eliminate these problems, and practice our fundamental freedom rightfully? To answer this question, we should understand the elements and the characteristic of the nature of freedom of expression, and the role of the actors whose duties and responsibilities are crucial in the prevalence of this fundamental freedom. To achieve this goal, this paper will explore the European practice to understand instructions found in the case-law of the European Court of Human rights for the rightful exercise of freedom of expression.

Keywords: collision of rights, European case-law, freedom opinion and expression, media law, freedom of information, online expression

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Authors: Dake Xiong, Ben Hankamer, Ian Ross

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The most urgent challenge of our time is to replace the depleting resources of fossil fuels by sustainable environmentally friendly alternatives. Hydrogen is a promising CO2-neutral fuel for a more sustainable future especially when produced photo-biologically. Hydrogen can be photosynthetically produced in unicellular green alga like Chlamydomonas reinhardtii, catalysed by the inducible highly active and bidirectional [FeFe]-hydrogenase enzymes (HydA). However, evolutionary and physiological constraints severely restrict the hydrogen yield of algae for industrial scale-up, mainly due to its competition among other metabolic pathways on photosynthetic electrons. Among them, a major challenge to be resolved is the inferior competitiveness of hydrogen production (catalysed by HydA) with NADPH production (catalysed by ferredoxin-NADP+-reductase (FNR)), which is essential for cell growth and takes up ~95% of photosynthetic electrons. In this work, the in vivo hydrogen production efficiency of mutants with ferredoxin-hydrogenase (Fd*-HydA1*) fusion protein construct, where the electron donor ferredoxin (Fd*) is fused to HydA1* and expressed in the model organism C. reinhardtii was investigated. Once Fd*-HydA1* fusion gene is expressed in algal cells, the fusion enzyme is able to draw the redistributed photosynthetic electrons and use them for efficient hydrogen production. From preliminary data, mutants with Fd*-HydA1* transgene showed a ~2-fold increase in the photosynthetic hydrogen production rate compared with its parental strain, which only possesses the native HydA in vivo. Therefore, a solid method of having more efficient hydrogen production in microalgae can be achieved through the expression of the synthetic enzymes.

Keywords: Chlamydomonas reinhardtii, ferredoxin, fusion protein, hydrogen production, hydrogenase

Procedia PDF Downloads 238

Authors: Duc Dong Do, Ngoc Ha Tran, Thanh Hai Dang, Cao Cuong Dang, Xuan Huan Hoang

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Global aligning two protein-protein interaction networks is an essentially important task in bioinformatics/computational biology field of study. It is a challenging and widely studied research topic in recent years. Accurately aligned networks allow us to identify functional modules of proteins and/ororthologous proteins from which unknown functions of a protein can be inferred. We here introduce a novel efficient heuristic global network alignment algorithm called FASTAn, including two phases: the first to construct an initial alignment and the second to improve such alignment by exerting a local optimization repeated procedure. The experimental results demonstrated that FASTAn outperformed the state-of-the-art global network alignment algorithm namely SPINAL in terms of both commonly used objective scores and the run-time.

Keywords: FASTAn, Heuristic algorithm, biological network alignment, protein-protein interaction networks

Procedia PDF Downloads 578

Authors: Xiao Zhou, Jianlin Cheng

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A single amino acid mutation can have a significant impact on the stability of protein structure. Thus, the prediction of protein stability change induced by single site mutations is critical and useful for studying protein function and structure. Here, we presented a deep learning network with the dropout technique for predicting protein stability changes upon single amino acid substitution. While using only protein sequence as input, the overall prediction accuracy of the method on a standard benchmark is >85%, which is higher than existing sequence-based methods and is comparable to the methods that use not only protein sequence but also tertiary structure, pH value and temperature. The results demonstrate that deep learning is a promising technique for protein stability prediction. The good performance of this sequence-based method makes it a valuable tool for predicting the impact of mutations on most proteins whose experimental structures are not available. Both the downloadable software package and the user-friendly web server (DNpro) that implement the method for predicting protein stability changes induced by amino acid mutations are freely available for the community to use.

Keywords: bioinformatics, deep learning, protein stability prediction, biological data mining

Procedia PDF Downloads 435

Authors: Chi-Ching Lee, Po-Jung Huang, Kuo-Yang Huang, Petrus Tang

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Background: Recent advances in high-throughput research technologies such as new-generation sequencing and multi-dimensional liquid chromatography makes it possible to dissect the complete transcriptome and proteome in a single run for the first time. However, it is almost impossible for many laboratories to handle and analysis these “BIG” data without the support from a bioinformatics team. We aimed to provide a web-based analysis platform for users with only limited knowledge on bio-computing to study the functional genomics and proteomics. Method: We use NCI-60 as an example dataset to demonstrate the power of the web-based analysis platform and data delivering system: C-eXpress takes a simple text file that contain the standard NCBI gene or protein ID and expression levels (rpkm or fold) as input file to generate a distribution map of gene/protein expression levels in a heatmap diagram organized by color gradients. The diagram is hyper-linked to a dynamic html table that allows the users to filter the datasets based on various gene features. A dynamic summary chart is generated automatically after each filtering process. Results: We implemented an integrated database that contain pre-defined annotations such as gene/protein properties (ID, name, length, MW, pI); pathways based on KEGG and GO biological process; subcellular localization based on GO cellular component; functional classification based on GO molecular function, kinase, peptidase and transporter. Multiple ways of sorting of column and rows is also provided for comparative analysis and visualization of multiple samples.

Keywords: cancer, visualization, database, functional annotation

Procedia PDF Downloads 592

Authors: Tesfaye Walle Mekonnen

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Maize is one of the most important staple food crops for most low-income households in the Sub-Saharan (SSA). Combined heat and drought stress is the major production threats that reduce the yield potential of biofortified maize and restrain various macro and micronutrient deficiencies highly prevalent in low-income people who rely solely on maize-based diets, SSA. This problem can be alleviated by crossing the biofortified inbred lines with different nutritional attributes, Fe, Zn, Protein, and Provitamin A, and developing agronomically superior and stable multi-nutrient maize of various genetic backgrounds. This aimed to understand the correlation between biofortified inbred lines per se and hybrid performance under combined heat and drought stress conditions (CSC). The experiment was conducted at CIMMYT, Zimbabwe, using α-lattice design with three replications. The hybrid effect was highly significant for zein fractions (α-, β-, γ- and δ-zein) zinc, (Zn), and iron (Fe) provitamin A, phytic acid, and grain yield. Under CSC, Fe, Zn concentration, provitamin A in grain and grain yield of hybrids were significantly decreased, however, the zein fraction content and phytic acid content increases in grain were increased under CSC. The phenotypic correlation between grain yield with Zn, Fe concentration, and Provitamin A in grain was strongly positive and higher under CSC than in well-watered conditions. The present investigation confirmed that under CSC, Fe, and Zn-enhanced hybrids could be forecasted to a certain scope based on the performance of and scientifically selected for desirable grain yield and related traits with CSC tolerance during hybrid development programs. In conclusion, the development of high-yielding and micronutrient-dense maize variety is possible under CSC, which could reduce the highly prevalent micronutrient in SSA.

Keywords: drought, Fe, heat, maize, protein, zein fractions, Zn

Procedia PDF Downloads 46

Authors: Duygu Çimen, Nilay Bereli, Adil Denizli

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In this study is to synthesis magnetic nanoparticles for purify protein C. For this aim, N-Methacryloyl-(L)-histidine methyl ester (MAH) containing 2-hydroxyethyl methacrylate (HEMA) based magnetic nanoparticles were synthesized by using micro-emulsion polymerization technique for templating protein C via metal chelation. The obtained nanoparticles were characterized with Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), zeta-size analysis and electron spin resonance (ESR) spectroscopy. After that, they were used for protein C purification from aqueous solution to evaluate/optimize the adsorption condition. Hereby, the effecting factors such as concentration, pH, ionic strength, temperature, and reusability were evaluated. As the last step, protein C was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Keywords: immobilized metal affinity chromatography (IMAC), magnetic nanoparticle, protein C, hydroxyethyl methacrylate (HEMA)

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Authors: N. Moradian, E. Timofeev, R. Tahir

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In this work, startability of the Busemann intake family with weak/strong conical shock, as most efficient intakes, via overboard mass spillage method is theoretically analyzed. Masterix and Candifix codes are used to numerically simulate few models of this type of intake and verify the theoretical results. Portions of the intake corresponding to various flow capture angles are considered to have mass spillage in the starting process of this intake. This approach allows for overboard mass spillage via a V-shaped slot with the tip of V coinciding with the focal point of the Busemann flow. The theoretical results, achieved using two different theories, of self-started Busemann takes with weak/strong conical shock show that significant improve in intake startability using overboard spillage technique. The starting phenomena of Busemann intakes with weak conical shock and seven different capture angles are numerically simulated at freestream Mach number of 3 to find the minimum area ratios of self-started intakes. The numerical results confirm the theoretical ones achieved by authors.

Keywords: Busemann intake, conical shock, overboard spillage, startability

Procedia PDF Downloads 187