Search results for: single cell protein (SCP)
8860 Cytotoxicity of a Short Chain Fatty Acid Histone Deactylase Inhibitor on HCT116 Human Colorectal Carcinoma Cell Line
Authors: N. A. Kazemi Sefat, M. M. Mohammadi, J. Hadjati, S. Talebi, M. Ajami, H. Daneshvar
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Colorectal cancer metastases result in a significant number of cancer related deaths. Histone deacetylase (HDAC) inhibitors induce growth arrest and apoptosis in a variety of human cancer cells. Sodium butyrate (SB) is a short chain fatty acid, belongs to HDAC inhibitors which is released in the colonic lumen as a consequence of fiber fermentation. In this study, we are about to assess the effect of sodium butyrate on HCT116 human colorectal carcinoma cell line. The viability of cells was measured by microscopic morphologic study and MTT assay. After 48 hours, treatments more than 10 mM lead to cell injury in HCT116 by increasing cell granulation and decreasing cell adhesion (p>0.05). After 72 hours, treatments at 10 mM and more lead to significant cell injury (p<0.05). Our results may suggest that the gene expression which is contributed in cell proliferation and apoptosis has been changed under pressure of HDAC inhibition.Keywords: colorectal cancer, sodium butyrate, cytotoxicity, MTT
Procedia PDF Downloads 3538859 Interaction between Kazal-Type Serine Proteinase Inhibitor SPIPm2 and Cyclophilin A from the Black Tiger Shrimp Penaeus monodon
Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit
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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon
Procedia PDF Downloads 2288858 The Effect of the Earthworm (Lumbricus rubellus) as the Source of Protein Feed and Pathogen Antibacterial for Broiler
Authors: Waode Nurmayani, Nikmatul Riswanda
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Broilers are chickens which are kept with the most efficient time and hoped get a good body weight. All things are done, for example with the improvement of feed and use antibiotics. Feed cost is the most cost to be spent. Nearly 80% of the cost is spent just for buy feed. Earthworm (Lumbricus rubellus) is a good choice to reduce the cost of feed protein source. The Earthworm has a high crude protein content of about 48.5%-61.9%, rich with proline amino acid about 15% of the 62 amino acids. Not only about protein, this earthworm also has a role in disease prevention. Prevention of disease in livestock usual with use feed supplement. Earthworm (Lumbricus rubellus) is one of the natural materials used as feed. In addition, several types of earthworms that have been known to contain active substances about antibacterial pathogens namely Lumbricus rubellus. The earthworm could be used as an antibiotic because it contain the antibody of Lumbricine active substance. So that, this animal feed from Lumbricus rubellus could improve the performance of broilers. Bioactive of anti-bacterial is called Lumbricine able to inhibit the growth of pathogenic bacteria in the intestinal wall so that the population of pathogenic bacteria is reduced. The method of write in this scientific writing is divided into 3 techniques, namely data completion, data analysis, and thinking pan from various literature about earthworm (Lumbricus rubellus) as broiler feed. It is expected that innovation of feed material of earthworm (Lumbricus rubellus) could reduce the cost of protein feed and the use of chemical antibiotics.Keywords: earthworm, broiler, protein, antibiotic
Procedia PDF Downloads 1488857 Apparent Ileal and Excreta Digestibility of Protein Poultry By-Product Meal in 21 to 28 Days of Age Broiler Chicken
Authors: N. Mahmoudnia, M. Khormali
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This experiment was conducted to determine the apparent protein digestibility of poultry byproduct meal (PBPM) from two industrial poultry slaughter-houses on Ross 308 male broiler chickens in independent comparisons. The experiment consisted of seven dietary treatments and three replicates per treatment with three broiler chickens per replicate in a completely randomized design. Dietary treatments consisted of a control corn- soybean diet, and levels 3, 6, and 9% PBPM produced by slaughter-house 1 and levels 3, 6, and 9% PBPM produced by slaughter house 2. Chromic oxide was added to the experimental diets as an indigestible marker. The apparent protein digestibility of each diet were determined with two methods of sample collection of ileum and excreta in 21-28 d of age. The results this experiment showed that use of PBPM had no significant effect on the performance of broiler chicks during period of experiments. The apparent protein digestibility of PBPM groups was significantly higher than control group by excreta sampling procedure (P<0.05). Using of PBPM 2 significantly (P<0.05) decreased the apparent protein digestibility values based on ileum sampling procedure vs control (85.21 vs. 90.14).Based results of this experiment,it is possible to use of PBPM 1 in broiler chicken.Keywords: poultry by-product meal, apparent protein digestibility, independed comparison, broiler chicken
Procedia PDF Downloads 4818856 Evaluation of Coagulation Efficiency of Protein Extracts from Lupinus Albus L., Moringa Stenopetala Cufod., Trigonella Foenum-Graecum L. And Vicia Faba L. For Water Purification
Authors: Neway Adele, Adey Feleke
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Access to clean drinking water is a basic human right. However, an estimated 1.2 billion people across the world consume unclean water daily. Interest has been growing in natural coagulants as the health and environmental concerns of conventional chemical coagulants are rising. Natural coagulants have the potential to serve as alternative water treatment agents. In this study, Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were evaluated as natural coagulants for water treatment. The protein extracts were purified from crude extracts using a protein purifier, and protein concentrations were determined by the spectrophotometric method. Small-volume coagulation efficiency tests were conducted on raw water taken from the Legedadi water treatment plant. These were done using a completely randomized design (CRD) experiment with settling times of 0 min (initial time), 90 min, 180 min and 270 min and protein extract doses of 5 mg/L, 10 mg/L, 15 mg/L and 20 mg/L. Raw water as negative control and polyelectrolyte as positive control were also included. The optical density (OD) values were measured for all the samples. At 270 min and 20 mg/L, the coagulation efficiency percentages for Lupinus albus, Moringa stenopetala, Trigonella foenum-graecum and Vicia faba protein extracts were 71%, 89%, 12% and 67% in the water sample collected in April 2019 respectively. Similarly, Lupinus albus, Moringa stenopetala and Vicia faba achieved 17%, 92% and 12% at 270 min settling times and 5 mg/L, 20 mg/L and 10 mg/L concentration in the water sample collected from August 2019, respectively. Negative control (raw water) and polyelectrolyte (positive control) were also 6 − 10% and 89 − 94% at 270 min settling time in April and August 2019, respectively. Among the four protein extracts, Moringa stenopetala showed the highest coagulation efficiency, similar to polyelectrolyte. This study concluded that Moringa stenopetala protein extract could be used as a natural coagulant for water purification in both sampling times.Keywords: coagulation efficiency, extraction, natural coagulant, protein extract
Procedia PDF Downloads 528855 Sider Bee Honey: Antitumor Effect in Some Experimental Tumor Cell Lines
Authors: Aliaa M. Issa, Mahmoud N. ElRouby, Sahar A. S. Ahmad, Mahmoud M. El-Merzabani
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Sider honey is a type of honey produced by bees feeding on the nectar of Sider tree, Ziziphus spina-christi (L) Desf . Honey is an effective agent for preventing, inhibiting and treating the growth of human and animal cancer cell lines in vitro and in vivo. The aim of the present study was to evaluate the impact of different dilutions from crude Sider honey and different duration times of exposure on the growth of six tumor cell lines (human cervical cancer cell line, HeLa; human hepatocellular carcinoma cell line, HepG-2; human larynx carcinoma cell line, Hep-2; brain tumor cell line, U251) as well as one animal cancerous cell line (Ehrlich ascites carcinoma cells line, EAC) and one normal cell line, Homo sapiens, human, (WISH) CCL-25. Different concentrations and treatment durations with Sider honey were tested on the growth of several cancer cell lines types. Histopathological changes in the tumor masses, animal survival, apoptosis and necrosis of the used cancer cell lines (using flow cytometry) were evaluated. Sider honey was administers either to the tumor mass itself by intratumoral injection or via drinking water. One-way ANOVA test was used for the analysis of (the means + standard error) of the optical density obtained from the Elisa reader and flow cytometry. The study revealed that different concentrations of Sider honey affected the growth patterns of all the studied cancer cell lines as well as their histopathological changes, and it depended on the cell line nature and the concentration of honey used. It is obvious that the relative animal survival percentage (bearing Ehrlich ascites carcinoma, EAC cells) was proportionally increased with the increase in the used honey concentrations. The study of apoptosis and necrosis using the flow cytometry technique emphasized the viability results. In conclusion, Sider honey was effective as antitumor agent, in the used concentrations.Keywords: antitumor, honey, sider, tumor cell lines
Procedia PDF Downloads 5268854 Modification of Escherichia coli PtolT Expression Vector via Site-Directed Mutagenesis
Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey
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Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression
Procedia PDF Downloads 3568853 Exploring Penicillin Resistance in Gonococcal Penicillin Binding Protein-2: Molecular Docking and Ligand Interaction Analysis
Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe
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Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids
Procedia PDF Downloads 588852 New Kinetic Approach to the Enzymatic Hydrolysis of Proteins: A Case of Thermolysin-Catalyzed Albumin
Authors: Anna Trusek-Holownia, Andrzej Noworyta
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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides
Procedia PDF Downloads 4308851 Assessment of Drug Delivery Systems from Molecular Dynamic Perspective
Authors: M. Rahimnejad, B. Vahidi, B. Ebrahimi Hoseinzadeh, F. Yazdian, P. Motamed Fath, R. Jamjah
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In this study, we developed and simulated nano-drug delivery systems efficacy in compare to free drug prescription. Computational models can be utilized to accelerate experimental steps and control the experiments high cost. Molecular dynamics simulation (MDS), in particular NAMD was utilized to better understand the anti-cancer drug interaction with cell membrane model. Paclitaxel (PTX) and dipalmitoylphosphatidylcholine (DPPC) were selected for the drug molecule and as a natural phospholipid nanocarrier, respectively. This work focused on two important interaction parameters between molecules in terms of center of mass (COM) and van der Waals interaction energy. Furthermore, we compared the simulation results of the PTX interaction with the cell membrane and the interaction of DPPC as a nanocarrier loaded by the drug with the cell membrane. The molecular dynamic analysis resulted in low energy between the nanocarrier and the cell membrane as well as significant decrease of COM amount in the nanocarrier and the cell membrane system during the interaction. Thus, the drug vehicle showed notably better interaction with the cell membrane in compared to free drug interaction with the cell membrane.Keywords: anti-cancer drug, center of mass, interaction energy, molecular dynamics simulation, nanocarrier
Procedia PDF Downloads 3328850 Assessment of Solar Hydrogen Production in Energetic Hybrid PV-PEMFC System
Authors: H. Rezzouk, M. Hatti, H. Rahmani, S. Atoui
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This paper discusses the design and analysis of a hybrid PV-Fuel cell energy system destined to power a DC load. The system is composed of a photovoltaic array, a fuel cell, an electrolyzer and a hydrogen tank. HOMER software is used in this study to calculate the optimum capacities of the power system components that their combination allows an efficient use of solar resource to cover the hourly load needs. The optimal system sizing allows establishing the right balance between the daily electrical energy produced by the power system and the daily electrical energy consumed by the DC load using a 28 KW PV array, a 7.5 KW fuel cell, a 40KW electrolyzer and a 270 Kg hydrogen tank. The variation of powers involved into the DC bus of the hybrid PV-fuel cell system has been computed and analyzed for each hour over one year: the output powers of the PV array and the fuel cell, the input power of the elctrolyzer system and the DC primary load. Equally, the annual variation of stored hydrogen produced by the electrolyzer has been assessed. The PV array contributes in the power system with 82% whereas the fuel cell produces 18%. 38% of the total energy consumption belongs to the DC primary load while the rest goes to the electrolyzer.Keywords: electrolyzer, hydrogen, hydrogen fueled cell, photovoltaic
Procedia PDF Downloads 4858849 YPFS Attenuating TH2 Cell-Mediated Allergic Inflammation by Regulating the TSLP Pathway
Authors: Xi Yu, Lili Gu, Huizhu Wang, Xiao Wei, Dandan Sheng, Xiaoyan Jiang, Min Hong
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Introduction: Hypersensitivity disease is difficult to cure completely because of its recurrence, yupingfengsan (YPFS) is used to treat the diseases with the advantage of reducing the recurrence,but the precise mechanism is not clear. Previous studies of our laboratory have shown that the extract of YPFS can inhibit Th2-type allergic contact dermatitis(ACD) induced by FITC.Besides, thymic stromal lymphopoietin(TSLP) have been proved to be a master switch for allergic inflammation. Based on these studies, we want to establish a mouse model of TSLP production based on Th2 cell-mediated allergic inflammation to explore the regulating mechanisms of YPFS on TSLP in Th2 cell-mediated allergic inflammation. Methods: Th2-type ACD mouse model: The mice were topically sensitized on the abdomens (induction phase) and elicited on its ears skin 6 day later (excitation phase) with FITC solution, and the ear swelling was measured to evaluate the allergic inflammation;A mouse model of TSLP production based on Th2 cell-mediated allergic inflammation (TSLP production model): the skin of the ear was sensitized on two consecutive days with FITC solution causing the production of TSLP;Mice were treated with YPFS extract,ELISA、Real-time PCR and Western-blotting were using to examine the mRNA and protein levels of TSLP\TSLPR and TLRs ect. Results: YPFS extract can attenuates Th2-type allergic inflammatory in mice;in TSLP production model, YPFS can inhibit the expression of TSLP、 TSLPR、TLRs and MyD88, So we deduce the possible mechanisms of YPFS to play a role of intervention is through TLRs- MyD88 dependent and independent pathway to reduce TSLP production.Keywords: YPFS, TSLP, TLRs, Th2-type allergic contact dermatitis
Procedia PDF Downloads 4168848 Prognostic and Predictive Value of Tumor: Infiltrating Lymphocytes in Triple Negative Breast Cancer
Authors: Wooseok Byon, Eunyoung Kim, Junseong Kwon, Byung Joo Song, Chan Heun Park
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Background/Purpose: Previous preclinical and clinical data suggest that increased lymphocytic infiltration would be associated with good prognosis and benefit from immunogenic chemotherapy especially in triple-negative breast cancer (TNBC). We investigated a single-center experience of TNBC and relationship with lymphocytic infiltration. Methods: From January 2004 to December 2012, at the Department of Surgery, Kangbuk Samsung Hospital, Sungkyunkwan University, School of Medicine, we retrospectively reviewed 897 breast cancer patients-clinical outcomes, clinicopathological characteristics, breast cancer subtypes. And we reviewed lymphocytic infiltration of TNBC specimens by two pathologists. Statistical analysis of risk factors associated with recurrence was performed. Results: A total of 897 patients, 76 were TNBC (8.47%). Mean age of TNBC patients were 50.95 (SD10.42) years, mean follow-up periods was 40.06 months. We reviewed 49 slides, and there were 8 recurrent breast cancer patients (16.32%), and 4 patients were expired (8.16%). There were 9 lymphocytic predominant breast cancers (LPBC)-carcinomas with either intratumoral lymphocytes in >60% of tumor cell nests. 1 patient of LPBC was recurred and 8 were not. In multivariate logistic regression, the odds ratio of lymphocytic infiltration was 0.59 (p=0.643). Conclusion: In a single-center experience of TNBC, the lymphocytic infiltration in tumor cell nest might be a good trend on the prognosis but there was not statistically significant.Keywords: tumor-infiltrating lymphocytes, triple negative breast cancer, medical and health sciences
Procedia PDF Downloads 3978847 Spectrophotometric Evaluation of Custom Microalgae-Based Bioink Formulations for Optimized Green Bioprinting
Authors: Olubusuyi Ayowole, Bashir Khoda
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Green bioprinting, from the context of merging 3D bioprinting with microalgae cell organization, holds promise for industrial-scale optimization. This study employs spectrophotometric analysis to explore post-bioprinting cell growth density variation within hybrid hydrogel biomaterial scaffolds. Three hydrogel biomaterials—Alginic acid sodium salt (ALGINATE), Nanofibrillated Cellulose (NFC) – TEMPO, and CarboxyMethyl Cellulose (CMC)—are chosen for their scaffolding capabilities. Bioink development and analysis of their impact on cell proliferation and morphology are conducted. Chlorella microalgae cell growth within hydrogel compositions is probed using absorbance measurements, with additional assessment of shear thinning properties. Notably, NFC exhibits reduced shear thinning compared to CMC. Results reveal that while mono-hydrogel substrates with pronounced adhesion inhibit Chlorella cell proliferation, Alginate fosters increased cell concentration alongside a slight viscosity rise.Keywords: green bioprinting, 3d bioprinting, microalgae cell, hybrid hydrogel scaffolds, spectrophotometric analysis, bioink development, shear thinning properties
Procedia PDF Downloads 148846 Effects of Cell Phone Usage on Psychological Health of Students
Authors: Avadhesh Kumar
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Background: The cell phone has rapidly become an integral, and, for some, an essential communication tool that is being used worldwide. Their use without any knowledge of their harmful effects like cancers and other health effects is not ‘quite’ safe. Studies on cancers due to electromagnetic radiations from cell phones are available, but there is a need to research on the detrimental physical and psychological effects on users like students. This study focused on certain psychological or mental health effects of cell phone usage amongst students. Materials and methods: The present study will be carried out on all the students of Banaras Hindu University. Students of both sexes from urban and rural backgrounds were selected at random and administered a pre- tested questionnaire which included aspects related to few common adverse psychological health signs and symptoms attributed to cell phone over-usage. Results: Stress was found to be the commonest symptom (51.47%) followed by irritability/anger (43.79%). Other common mental symptoms included lack of concentration and academic performance, insomnia, anxiety etc. Suggestions: This study confirms that the younger generation, who are the most frequent cell phone users, needs to be aware of the adverse health effects of cell phone usage especially the mental aspects and take preventive measures to minimize and control the same. Less dependence on the device, a curtailing time period spent on talking, communicating more by texting, etc. are some of the practical measures suggested.Keywords: cell phones, psychological health effects, students, mental health
Procedia PDF Downloads 3038845 Cytotoxic Effect of Purified and Crude Hyaluronidase Enzyme on Hep G2 Cell Line
Authors: Furqan M. Kadhum, Asmaa A. Hussein, Maysaa Ch. Hatem
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Hyaluronidase enzyme was purified from the clinical isolate Staphyloccus aureus in three purification steps, first by precipitation with 90% saturated ammonium sulfate, ion exchange chromatography on DEAE-Cellulose, and gel filtration chromatography throughout Sephacryl S-300. Specific activity of the purified enzyme was reached 930 U/mg protein with 7.4 folds of purification and 46.5% recovery. The enzyme has an average molecular weight of about 69 kDa, with an optimum pH of enzyme activity and stability at pH 7, also the optimum temperature for activity was 37oC. The enzyme was stable with full activity at a temperature ranged between 30-40 oC. Metal ions showed variable inhibitory degree with the strongest effect for Fe+3, however, the chelating and reducing agents had no or little effects. Cytotoxic studies for purified and crude hyaluronidase against cancer cell Hep G2 type at different enzyme concentrations and exposure times showed that the inhibition effect of both crude and purified enzyme increased by increasing the enzyme concentration with no change was observed at 24hr, while at 48 and 72 hrs the same inhibition rate were observed for purified enzyme and differ for the crude filtrate.Keywords: hyaluronidase, S. aureus, metal ions, cytotoxicity
Procedia PDF Downloads 4388844 Transition to Hydrogen Cities in Korea and Japan
Authors: Minhee Son, Kyung Nam Kim
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This study explores the plan of the Korean and Japanese governments to transition into the hydrogen economy. Two motor companies, Hyundai Motor Company from Korea and Toyota from Japan, released the Hydrogen Fuel Cell Vehicle to monopolize the green energy automobile market. Although, they are the main countries which emit greenhouse gas, hydrogen energy can bring from a certain industry places, such as chemical plants and steel mills. Recent, the two countries have been focusing on the hydrogen industry including a fuel cell vehicle, a hydrogen station, a fuel cell plant, a residential fuel cell. The purpose of this paper is to find out the differences of the policies in the two countries to be hydrogen societies. We analyze the behavior of the public and private sectors in Korea and Japan about hydrogen energy and fuel cells for the transition of the hydrogen economy. Finally we show the similarities and differences of both countries in hydrogen fuel cells. And some cities have feature such as Hydrogen cities. Hydrogen energy can make impact environmental sustainability.Keywords: fuel cell, hydrogen city, hydrogen fuel cell vehicle, hydrogen station, hydrogen energy
Procedia PDF Downloads 4738843 Design of a Controlled BHJ Solar Cell Using Modified Organic Vapor Spray Deposition Technique
Authors: F. Stephen Joe, V. Sathya Narayanan, V. R. Sanal Kumar
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A comprehensive review of the literature on photovoltaic cells has been carried out for exploring the better options for cost efficient technologies for future solar cell applications. Literature review reveals that the Bulk Heterojunction (BHJ) Polymer Solar cells offer special opportunities as renewable energy resources. It is evident from the previous studies that the device fabricated with TiOx layer shows better power conversion efficiency than that of the device without TiOx layer. In this paper, authors designed a controlled BHJ solar cell using a modified organic vapor spray deposition technique facilitated with a vertical-moving gun named as 'Stephen Joe Technique' for getting a desirable surface pattern over the substrate to improving its efficiency over the years for industrial applications. We comprehended that the efficient processing and the interface engineering of these solar cells could increase the efficiency up to 5-10 %.Keywords: BHJ polymer solar cell, photovoltaic cell, solar cell, Stephen Joe technique
Procedia PDF Downloads 5338842 FTIR Spectroscopy for in vitro Screening in Microbial Biotechnology
Authors: V. Shapaval, N. K. Afseth, D. Tzimorotas, A. Kohler
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Globally there is a dramatic increase in the demand for food, energy, materials and clean water since natural resources are limited. As a result, industries are looking for ways to reduce rest materials and to improve resource efficiency. Microorganisms have a high potential to be used as bio factories for the production of primary and secondary metabolites that represent high-value bio-products (enzymes, polyunsaturated fatty acids, bio-plastics, glucans, etc.). In order to find good microbial producers, to design suitable substrates from food rest materials and to optimize fermentation conditions, rapid analytical techniques for quantifying target bio products in microbial cells are needed. In the EU project FUST (R4SME, Fp7), we have developed a fully automated high-throughput FUST system based on micro-cultivation and FTIR spectroscopy that facilitates the screening of microorganisms, substrates and fermentation conditions for the optimization of the production of different high-value metabolites (single cell oils, bio plastics). The automated system allows the preparation of 100 samples per hour. Currently, The FUST system is in use for screening of filamentous fungi in order to find oleaginous strains with the ability to produce polyunsaturated fatty acids, and the optimization of cheap substrates, derived from food rest materials, and the optimization of fermentation conditions for the high yield of single cell oil.Keywords: FTIR spectroscopy, FUST system, screening, biotechnology
Procedia PDF Downloads 4358841 Effect of Different Processing Methods on the Quality Attributes of Pigeon Pea Used in Bread Production
Authors: B. F. Olanipekun, O. J. Oyelade, C. O. Osemobor
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Pigeon pea is a very good source of protein and micronutrient, but it is being underutilized in Nigeria because of several constraints. This research considered the effect of different processing methods on the quality attributes of pigeon pea used in bread production towards enhancing its utility. Pigeon pea was obtained at a local market and processed into the flour using three processing methods: soaking, sprouting and roasting and were used to bake bread in different proportions. Chemical composition and sensory attributes of the breads were thereafter determined. The highest values of protein and ash contents were obtained from 20 % substitution of sprouted pigeon pea in wheat flour and may be attributable to complex biochemical changes occurring during hydration, to invariably lead to protein constituent being broken down. Hydrolytic activities of the enzymes from the sprouted sample resulted in improvement in the constituent of total protein probably due to reduction in the carbohydrate content. Sensory qualities analyses showed that bread produced with soaked and roasted pigeon pea flours at 5 and 10% inclusion, respectively were mostly accepted than other blends, and products with sprouted pigeon pea flour were least accepted. The findings of this research suggest that supplementing wheat flour with sprouted pigeon peas have more nutritional potentials. However, with sensory analysis indices, the soaked and roasted pigeon peas up to 10% are majorly accepted, and also can improve the nutritional status. Overall, this will be very beneficial to population dependent on plant protein in order to combat malnutrition problems.Keywords: pigeon pea, processing, protein, malnutrition
Procedia PDF Downloads 2418840 Cancer Stem Cell-Associated Serum Proteins Obtained by Maldi TOF/TOF Mass Spectrometry in Women with Triple-Negative Breast Cancer
Authors: Javier Enciso-Benavides, Fredy Fabian, Carlos Castaneda, Luis Alfaro, Alex Choque, Aparicio Aguilar, Javier Enciso
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Background: The use of biomarkers in breast cancer diagnosis, therapy, and prognosis has gained increasing interest. Cancer stem cells (CSCs) are a subpopulation of tumor cells that can drive tumor initiation and may cause relapse. Therefore, due to the importance of diagnosis, therapy, and prognosis, several biomarkers that characterize CSCs have been identified; however, in treatment-naïve triple-negative breast tumors, there is an urgent need to identify new biomarkers and therapeutic targets. According to this, the aim of this study was to identify serum proteins associated with cancer stem cells and pluripotency in women with triple-negative breast tumors in order to subsequently identify a biomarker for this type of breast tumor. Material and Methods: Whole blood samples from 12 women with histopathologically diagnosed triple-negative breast tumors were used after obtaining informed consent from the patient. Blood serum was obtained by conventional procedure and frozen at -80ºC. Identification of cancer stem cell-associated proteins was performed by matrix-assisted laser desorption/ionisation-assisted laser desorption/ionisation mass spectrometry (MALDI-TOF MS), protein analysis was obtained using the AB Sciex TOF/TOF™ 5800 system (AB Sciex, USA). Sequences not aligned by ProteinPilot™ software were analyzed by Protein BLAST. Results: The following proteins related to pluripotency and cancer stem cells were identified by MALDI TOF/TOF mass spectrometry: A-chain, Serpin A12 [Homo sapiens], AIEBP [Homo sapiens], Alpha-one antitrypsin, AT {internal fragment} [human, partial peptide, 20 aa] [Homo sapiens], collagen alpha 1 chain precursor variant [Homo sapiens], retinoblastoma-associated protein variant [Homo sapiens], insulin receptor, CRA_c isoform [Homo sapiens], Hydroxyisourate hydrolase [Streptomyces scopuliridis], MUCIN-6 [Macaca mulatta], Alpha-actinin-3 [Chrysochloris asiatica], Polyprotein M, CRA_d isoform, partial [Homo sapiens], Transcription factor SOX-12 [Homo sapiens]. Recommendations: The serum proteins identified in this study should be investigated in the exosome of triple-negative breast cancer stem cells and in the blood serum of women without breast cancer. Subsequently, proteins found only in the blood serum of women with triple-negative breast cancer should be identified in situ in triple-negative breast cancer tissue in order to identify a biomarker to study the evolution of this type of cancer, or that could be a therapeutic target. Conclusions: Eleven cancer stem cell-related serum proteins were identified in 12 women with triple-negative breast cancer, of which MUCIN-6, retinoblastoma-associated protein variant, transcription factor SOX-12, and collagen alpha 1 chain are the most representative and have not been studied so far in this type of breast tumor. Acknowledgement: This work was supported by Proyecto CONCYTEC–Banco Mundial “Mejoramiento y Ampliacion de los Servicios del Sistema Nacional de Ciencia Tecnología e Innovacion Tecnologica” 8682-PE (104-2018-FONDECYT-BM-IADT-AV).Keywords: triple-negative breast cancer, MALDI TOF/TOF MS, serum proteins, cancer stem cells
Procedia PDF Downloads 2028839 Catalytic Study of Natural Gas Based Solid Oxide Fuel Cell
Authors: Nasir Iqbal, Khurram Siraj, Rizwan Raza
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Solid oxide fuel cell (SOFC) is the promising technology now days. SOFC can be operated with different types of fuels available. In this work catalytic anode is prepared with metal oxides i.e. Li, Ni, Zn and Sn and tested for catalytic activity with natural gas as a fuel. The operating temperature range is 170-750°C as observed with the help of TGA. Electrical conductivity and fuel cell performance has been observed for four different samples with varying composition of Sn and Zn. It is concluded that the sample having greater concentration of Zn shows better conductivity and power density results. All the results are promising and verified with different characterizations.Keywords: catalytic activity, solid oxide fuel cell, energy material, natural gas
Procedia PDF Downloads 658838 Proteomic Analysis of Cytoplasmic Antigen from Brucella canis to Characterize Immunogenic Proteins Responded with Naturally Infected Dogs
Authors: J. J. Lee, S. R. Sung, E. J. Yum, S. C. Kim, B. H. Hyun, M. Her, H. S. Lee
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Canine brucellosis is a critical problem in dogs leading to reproductive diseases which are mainly caused by Brucella canis. There are, nonetheless, not clear symptoms so that it may go unnoticed in most of the cases. Serodiagnosis for canine brucellosis has not been confirmed. Moreover, it has substantial difficulties due to broad cross-reactivity between the rough cell wall antigens of B. canis and heterospecific antibodies present in normal, uninfected dogs. Thus, this study was conducted to characterize the immunogenic proteins in cytoplasmic antigen (CPAg) of B. canis, which defined the antigenic sensitivity of the humoral antibody responses to B. canis-infected dogs. In analysis of B. canis CPAg, first, we extracted and purified the cytoplasmic proteins from cultured B. canis by hot-saline inactivation, ultrafiltration, sonication, and ultracentrifugation step by step according to the sonicated antigen extract method. For characterization of this antigen, we checked the sort and range of each protein on SDS-PAGE and verified the immunogenic proteins leading to reaction with antisera of B. canis-infected dogs. Selected immunodominant proteins were identified using MALDI-MS/MS. As a result, in an immunoproteomic assay, several polypeptides in CPAg on one or two-dimensional electrophoresis (DE) were specifically reacted to antisera from B. canis-infected dogs but not from non-infected dogs. The polypeptides with approximate 150, 80, 60, 52, 33, 26, 17, 15, 13, 11 kDa on 1-DE were dominantly recognized by antisera from B. canis-infected dogs. In the immunoblot profiles on 2-DE, ten immunodominant proteins in CPAg were detected with antisera of infected dogs between pI 3.5-6.5 at approximate 35 to 10 KDa, without any nonspecific reaction with sera in non-infected dogs. Ten immunodominant proteins identified by MALDI-MS/MS were identified as superoxide dismutase, bacteroferritin, amino acid ABC transporter substrate-binding protein, extracellular solute-binding protein family3, transaldolase, 26kDa periplasmic immunogenic protein, Rhizopine-binding protein, enoyl-CoA hydratase, arginase and type1 glyceraldehyde-3-phosphate dehydrogenase. Most of these proteins were determined by their cytoplasmic or periplasmic localization with metabolism and transporter functions. Consequently, this study discovered and identified the prominent immunogenic proteins in B. canis CPAg, highlighting that those antigenic proteins may accomplish a specific serodiagnosis for canine brucellosis. Furthermore, we will evaluate those immunodominant proteins for applying to the advanced diagnostic methods with high specificity and accuracy.Keywords: Brucella canis, Canine brucellosis, cytoplasmic antigen, immunogenic proteins
Procedia PDF Downloads 1378837 Effect of Many Levels of Undegradable Protein on Performance, Blood Parameters, Colostrum Composition and Lamb Birth Weight in Pregnant Ewes
Authors: Maria Magdy Danial Riad
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The objective of this study was to investigate the effect of different protein sources with different degradability ratios during late gestation of ewes on colostrum composition and its IgG concentration, body weight change of dams, and birth weight of their lambs. Objectives: 35 multiparous native crossbred ewes (BW= 59±2.5kg) were randomly allocated to five dietary treatments (7 ewes / treatment) for 2 months prior to lambing. Methods: Experimental diets were isonitrogenous (12.27% CP) and isocaloric (2.22 Mcal ME/kg DM). In diet I (the control), solvent extract soybeans (SESM 33% RUP of CP), II feed grade urea (FGU 31% RUP), III slow release urea (SRU 31% RUP). As sources of undegradable protein, extruded expeller SBM-EESM 40 (37% RUP) and extruded expeller SBM-EESM 60 (41% RUP) were used in groups IV and V, respectively. Results showed no significant effect on feed intake, crude protein (CP), metabolizable energy (ME), and body condition score (BCS). Ewes fed the 37% RUP diet gained more (p<0.05) weight compared with ewes fed the 31% RUP diet (5.62 vs. 2.5kg). Ewes in EESM 60 had the highest levels of fat, protein, total solid, solid not fat, and immunoglobulin and the lowest in urea N content (P< 0.05) in colostrum during the first 24hrs after lambing. Conclusions: Protein source and RUP levels in ewes’ diets had no significant effect (P< 0.05) on lambs’ birth weight and ewes' blood biochemical parameters. Increasing the RUP content of diet during late gestation resulted in an increase in colostrum constituents and its IgG level but had no effect on ewes’ performance and their lambs’ outcome.Keywords: colostrum, ewes, lambs output, pregnancy, undegradable protein
Procedia PDF Downloads 408836 Continuum-Based Modelling Approaches for Cell Mechanics
Authors: Yogesh D. Bansod, Jiri Bursa
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The quantitative study of cell mechanics is of paramount interest since it regulates the behavior of the living cells in response to the myriad of extracellular and intracellular mechanical stimuli. The novel experimental techniques together with robust computational approaches have given rise to new theories and models, which describe cell mechanics as a combination of biomechanical and biochemical processes. This review paper encapsulates the existing continuum-based computational approaches that have been developed for interpreting the mechanical responses of living cells under different loading and boundary conditions. The salient features and drawbacks of each model are discussed from both structural and biological points of view. This discussion can contribute to the development of even more precise and realistic computational models of cell mechanics based on continuum approaches or on their combination with microstructural approaches, which in turn may provide a better understanding of mechanotransduction in living cells.Keywords: cell mechanics, computational models, continuum approach, mechanical models
Procedia PDF Downloads 3548835 Investigation of The Effects of Hydroxytyrosol on Cytotoxicity, Apoptosis, PI3K/Akt, and ERK 1/2 Pathways in Ovarian Cancer Cell Cultures
Authors: Latife Merve Oktay, Berrin Tugrul
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Hydroxytyrosol (HT) is a phenolic phytochemical molecule derived from the hydrolysis of oleuropein, which originates during the maturation of the olives. It has recently received particular attention because of its antioxidant, anti-proliferative, pro-apoptotic and anti-inflammatory activities. In this study, we investigated the cytotoxic and apoptotic effects of hydroxytyrosol and its effects on phosphatidylinositol 3-kinase/Akt (PI3K/Akt) and extracellular signal-regulated kinase 1/2 (ERK 1/2) signaling pathways in human ovarian cancer cell lines OVCAR-3 and MDAH-2774. XTT cell proliferation kit, Cell Death Detection Elisa Plus Kit (Roche) and Human Apoptosis Array (R&D Systems) were used to determine the cytotoxic and apoptotic effects of HT in OVCAR-3 and MDAH-2774 cell lines at 24, 48, 72, and 96 h. Effect of HT on PI3K/Akt and ERK 1/2 signaling pathways were investigated by using specific inhibitors of these pathways. IC50 values of HT were found to be 102.3 µM in MDAH-2774 cells at 72 h and 51.5 µM in OVCAR-3 cells at 96 h. Apoptotic effect of HT in MDAH-2774 cells was the highest at 50 µM at 72 h, and kept decreasing at 100 and 150 µM concentrations and was not seen at 200 µM and higher concentrations. Highest apoptotic effect was seen at 100 µM concentration in OVCAR-3 cells at 96 h, however apoptotic effect was decreased over 100 µM concentrations. According to antibody microarray results, HT increased the levels of pro-apoptotic molecules Bad, Bax, active caspase-3, Htra2/Omi by 2.0-, 1.4-, 1.2-, 4.2-fold, respectively and also increased the levels of pro-apoptotic death receptors TRAIL R1/DR4, TRAIL R2/DR5, FAS/TNFRSF6 by 2.1-, 1.7-, 1.6-fold, respectively, however, it decreased the level of Survivin by 1.6-fold which is one of the inhibitor of apoptosis protein (IAP) family in MDAH-2774 cells. In OVCAR-3 cells, HT decreased the levels of anti-apoptotic proteins Bcl-2, pro-caspase 3 by 3.1-, 8.2-fold, respectively and IAP family proteins CIAP-1, CIAP-2, XIAP, Livin, Survivin by 6.5-, 6.0-, 3.2-, 2.2-, 2.7-fold, respectively and increased the level of cytochrome-c by 1.2-fold. We have shown that HT shows its cytotoxic and apoptotic effect through inhibiting ERK 1/2 signaling pathway in both OVCAR-3 and MDAH-2774 cells. Further studies are needed to investigate molecular mechanisms and modulatory effects of hydroxytyrosol.Keywords: apoptosis, cytotoxicity, hydroxytyrosol, ovarian cancer
Procedia PDF Downloads 3498834 Analysis of Performance of 3T1D Dynamic Random-Access Memory Cell
Authors: Nawang Chhunid, Gagnesh Kumar
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On-chip memories consume a significant portion of the overall die space and power in modern microprocessors. On-chip caches depend on Static Random-Access Memory (SRAM) cells and scaling of technology occurring as per Moore’s law. Unfortunately, the scaling is affecting stability, performance, and leakage power which will become major problems for future SRAMs in aggressive nanoscale technologies due to increasing device mismatch and variations. 3T1D Dynamic Random-Access Memory (DRAM) cell is a non-destructive read DRAM cell with three transistors and a gated diode. In 3T1D DRAM cell gated diode (D1) acts as a storage device and also as an amplifier, which leads to fast read access. Due to its high tolerance to process variation, high density, and low cost of memory as compared to 6T SRAM cell, it is universally used by the advanced microprocessor for on chip data and program memory. In the present paper, it has been shown that 3T1D DRAM cell can perform better in terms of fast read access as compared to 6T, 4T, 3T SRAM cells, respectively.Keywords: DRAM Cell, Read Access Time, Retention Time, Average Power dissipation
Procedia PDF Downloads 3048833 Investigation of Ezetimibe Administration on Cell Survival Markers in Kidney Ischemia
Authors: Zahra Heydari
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Introduction: One of the major clinical issues is acute renal failure, which is caused by ischemia-reperfusion of the kidney and is associated with high mortality. Despite advances in this area, important issues such as tissue necrosis, cell apoptosis, and so on in damaged tissue are suggestive for more researches and study on this subject. Objective: Evaluation of the potential utility of Ezetimibe in reducing injuries and cell death induced by kidney ischemia/ reperfusion through inducing expression changes of different cellular pathways in adult Sprague-Dawley rats. Materials and methods: Forty rats weighing 180-200g were divided into 4 groups. For this purpose, the first right kidneys of the rats were removed during surgery. After 20 days, the left renal artery was closed with a soft clamp and reperfusion was performed. After 24 hours, blood samples were collected and sent to the laboratory with kidneys to measure bax and bcl-2 by Western blotting and histopathological tests. Results: Quantitative damage reviews of Kidney tissue indicates damage Acute and severe tubular lesions were observed in the ischemia group. Also, the amount of injury was significantly reduced in the treatment group. There was also a significant difference between the ischemia and sham groups. In general, the results show that a single dose of 1.2 mg/kg of ezetimibe can reduce the bax/ bcl-2 ratio compared to the ischemia group. In general, the results showed Ezetimibe is effective in reducing cell damage and death due to ischemia/ reperfusion after renal ischemia through changes in the expression of various cellular pathways in rats.Keywords: acute renal failure, renal ischemia-reperfusion injury, ezetimibe, apoptosis
Procedia PDF Downloads 1838832 Facile Synthesis of Novel Substituted Aryl-Thiazole (SAT) Analogs via One-Pot Multicomponent Reaction as Potent Cytotoxic Agents against Cancer Cell Lines
Authors: Salma Mirza, Syeda Asma Naqvi, Khalid Mohammed Khan, M. Iqbal Choudhary
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In this study twenty-five (25) newly synthesized compounds substituted aryl thiazoles (SAT) 1-25 were synthesized, and in vitro cytotoxicity of these compounds was evaluated against four cancer cell lines namely, MCF-7 (ER+ve breast), MDA-MB-231 (ER-ve breast), HCT116 (colorectal), and, HeLa (cervical) and compared with the standard anticancer drug doxorubicin with IC50 value of 1.56 ± 0.05 μM. Among them, compounds 1, 4-8 and 19 were found to be active against all four cell lines. Compound 20 was found to be selectively active against MCF7 cells with IC50 value of 40.21 ± 4.15 µM, whereas compound 19 was active against only MCF7 and HeLa cells with IC50 values of 46.72 ± 1.8 and 19.86 ± 0.11 μM, respectively. These results suggest that aryl thiazoles 1 and 4 deserve to be investigated further in vivo as anti-cancer agents.Keywords: anticancer agents, breast cancer cell lines (MCF7, MDA-MB-231), colorectal cancer cell line (HCT-116), cervical cancer cell line (HeLa), Thiazole derivatives
Procedia PDF Downloads 2918831 Immunoinformatic Design and Evaluation of an Epitope-Based Tetravalent Vaccine against Human Hand, Foot, and Mouth Disease
Authors: Aliyu Maje Bello, Yaowaluck Maprang Roshorm
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Hand, foot, and mouth disease (HFMD) is a highly contagious viral infection affecting mostly infants and children. Although the Enterovirus A71 (EV71) is usually the major causative agent of HFMD, other enteroviruses such as coxsackievirus A16, A10, and A6 are also found in some of the recent outbreaks. The commercially available vaccines have demonstrated their effectiveness against only EV71 infection but no protection against other enteroviruses. To address the limitation of the monovalent EV71 vaccine, the present study thus designed a tetravalent vaccine against the four major enteroviruses causing HFMD and primarily evaluated the designed vaccine using an immunoinformatics approach. The immunogen was designed to contain the EV71 VP1 protein and multiple reported epitopes from all four distinct enteroviruses and thus designated a tetravalent vaccine. The 3D structure of the designed tetravalent vaccine was modeled, refined, and validated. Epitope screening showed the presence of B-cell, CTL, CD4 T cell, and IFN epitopes with vast application among the Asian population. Docking analysis confirmed the stable and strong binding interactions between the immunogen and immune receptor B-cell receptor (BCR). In silico cloning and immune simulation analyses guaranteed high efficiency and sufficient expression of the vaccine candidate in humans. Overall, the promising results obtained from the in-silico studies of the proposed tetravalent vaccine make it a potential candidate worth further experimental validation.Keywords: enteroviruses, coxsackieviruses, hand foot and mouth disease, immunoinformatics, tetravalent vaccine
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