Search results for: multidrug resistance protein 3

Authors: Y. V. S. Annapurna, V. V. Lakshmi

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Urinary tract infection (UTI) is one of the most common infectious diseases at the community level with a high rate of morbidity . This is further augmented by increase in the number of resistant and multi resistant strains of bacteria particularly by those producing Extended spectrum of beta lactamases. The present study was aimed at analysis of antibiograms of E.coli and Klebsiella sp causing urinary tract infections. Between November 2011 and April 2013, a total of 1120 urine samples were analyzed,. Antibiotic sensitivity testing was done with 542(48%) isolates of E.coli and 446(39%) of Klebsiella sp using the standard disc diffusion method against eleven commonly used antibiotics .Organisms showed high susceptibility to Amikacin and Netilimicin and low susceptibility to Cephalosporins. MAR index was calculated for the multidrug resistant strains. Maximum MAR index detected among the isolates was 0.9. Phenotypic identification for ESBL production was confirmed by double disk synergy test (DDST) according to CLSI guidelines. Plasmid profile of the isolates was carried out using alkaline hydrolysis method. Agarose-gel electrophoresis showed presence of high-molecular weight plasmid DNA among the ESBL strains. This study emphasizes the importance of indiscriminate use of antibiotics which if discontinued, in turn would prevent further development of bacterial drug resistance. For this, a proper knowledge of susceptibility pattern of uropathogens is necessary before prescribing empirical antibiotic therapy and it should be made mandatory.

Keywords: escherichia coli, extended spectrum of beta lactamase, Klebsiella spp, Uropathogens

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Authors: Daina Jonkus, Solvita Petrovska, Dace Smiltina, Lasma Cielava

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The beta-lactoglobulin and kappa-casein are milk proteins which are important for milk composition. Cows with beta-lactoglobulin and kappa-casein gene BB genotypes have highest milk crude protein and fat content. The aim of the study was to determinate the frequencies of milk protein gene polymorphisms in local Latvian Brown (LB) cows breed and analyze the influence of beta-lactoglobulin and kappa-casein genotypes to milk productivity traits. 102 cows’ genotypes of milk protein genes were detected using Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) and electrophoresis on 3% agarose gel. For beta-lactoglobulin were observed 2 types of alleles A and B and for kappa-casein 3 types: A, B and E. Highest frequency in beta-lactoglobulin gene was observed for B allele – 0.926. Molecular analysis of beta-lactoglobulin gene shows 86.3% of individuals are homozygous by B allele and animals are with genotypes BB and 12.7% of individuals are heterozygous with genotypes AB. The highest milk yield 4711.7 kg was for 1st lactation cows with AB genotypes, whereas the highest milk protein content (3.35%) and fat content (4.46 %) was for BB genotypes. Analysis of the kappa-casein locus showed a prevalence of the A allele – 0.750. The genetic variant of B was characterized by a low frequency – 0.240. Moreover, the frequency of E occurred in the LB cows’ population with very low frequency – 0.010. 54.9 % of cows are homozygous with genotypes AA, and only 4.9 % are homozygous with genotypes BB. 32.8 % of individuals are heterozygous with genotypes AB, and 2.0 % are with AE. The highest milk productivity was for 1st lactation cows with AB genotypes: milk yield 4620.3 kg, milk protein content 3.39% and fat content 4.53 %. According to the results, in local Latvian brown there are only 2.9% of cows are with BB-BB genotypes, which is related to milk coagulation ability and affected cheese production yield. Acknowledgment: the investigation is supported by VPP 2014-2017 AgroBioRes Project No. 3 LIVESTOCK.

Keywords: beta-lactoglobulin, cows, genotype frequencies, kappa-casein

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Authors: Purva Mishra, Rajesh Potlia, Kuljeet Singh Sandhu

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The role of glycyl residues in the protein structure has lingered within the research community for the last several decades. Glycyl residue is the only amino acid that is achiral due to the lack of a side chain and can, therefore, exhibit Ramachandran conformations that are disallowed for L-amino acids. The structural and functional significance of glycyl residues with L-disallowed conformation, however, remains obscure. Through statistical analysis of various datasets, we found that the glycyls with L-disallowed conformations are over-represented at disease-associated sites and tend to be evolutionarily conserved. The mutations of L-disallowed glycyls tend to destabilize the native conformation, reduce protein solubility, and promote inter-molecular aggregations. We uncovered a structural motif referred to as “β-crescent” formed around the L-disallowed glycyl, which prevents β-sheet aggregation by disrupting the alternating pattern of β-pleats. The L-disallowed conformation of glycyls also holds predictive power to infer the pathogenic missense variants. Altogether, our observations highlight that the L-disallowed conformation of glycyls is selected to facilitate native folding and prevent inter-molecular aggregations. The findings may also have implications for designing more stable proteins and prioritizing the genetic lesions implicated in diseases.

Keywords: Ramachandran plot, β-sheet, protein stability, protein aggregation

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Authors: Mohamed M. Alsull

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In the present study, two microalgae species “Nannochloropsis sp. and Tetraselmis sp.” isolated from Penang National Park coastal waters, Malaysia; were cultivated under combined various laboratory conditions “salinity, light, nitrogen limitation and starvation”. Growth rate, dry weight, chlorophyll a content, total lipid and protein contents, were estimated at mid exponential growth phase. Both Nannochloropsis sp. and Tetraselmis sp. showed remarkable decrease in growth rate, chlorophyll a content and protein content companied with increase in lipid content under nitrogen limitation and starvation conditions. Maintaining Nannochloropsis sp. under salinity 15‰ caused only significant decrease in total protein content; while Tetraselmis sp. grown at the same salinity caused decrease in the growth rate, chlorophyll a, dry weight and total protein content only when nitrogen was available.

Keywords: biochemical composition, light, microalgae, nitrogen limitation, salinity

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Authors: Sena Su, Burak Ozbek, Yesim M. Sahin, Sevil Yucel, Dilek Kazan, Faik N. Oktar, Nazmi Ekren, Oguzhan Gunduz

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In recent years, protein, lipid or polysaccharide-based polymers have been used in order to develop biodegradable materials and their chemical nature determines the physical properties of the resulting films. Among these polymers, proteins from different sources have been extensively employed because of their relative abundance, film forming ability, and nutritional qualities. In this study, the biodegradable composite nanofiber films based on fish sarcoplasmic protein (FSP) were prepared via electrospinning technique. Biodegradable polycaprolactone (PCL) was blended with the FSP to obtain hybrid FSP/PCL nanofiber mats with desirable physical properties. Mixture solutions of FSP and PCL were produced at different concentrations and their density, viscosity, electrical conductivity and surface tension were measured. Mechanical properties of electrospun nanofibers were evaluated. Morphology of composite nanofibers was observed using scanning electron microscopy (SEM). Moreover, Fourier transform infrared spectrometer (FTIR) studies were used for analysis chemical composition of composite nanofibers. This study revealed that the FSP based nanofibers have the potential to be used for different applications such as biodegradable packaging, drug delivery, and wound dressing, etc.

Keywords: edible film, electrospinning, fish sarcoplasmic protein, nanofiber

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Authors: Hawbash Jalil

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In this study, the effects of whey protein based films on various properties of kashar cheese were examined. In the study, edible film solutions based on whey protein isolate, whey protein isolate + transglutaminase enzyme and whey protein isolate + chitosan were produced and Kashar cheese samples were coated with these films by dipping method and stored at +4 ºC for 60 days. Chemical, microbiological and textural analyzes were carried out on samples at 0, 30 and 60 days of storage. As a result of the study, the highest dry matter and total nitrogen values were obtained from uncoated control samples This is an indication that the coatings limit water vapor permeability. The highest acidity and pH values obtained from the samples as storage results were 3.33% and 5.86%, respectively, in the control group samples. Both acidity and pH rise in these groups, is a consequence of the buffering of pH changes of hydrolsis products which are as a result of proteolysis occurring in the sample. Nitrogen changes and lipolysis values, which are indicative of the degree of hydrolysis of proteins and triglycerides in kashar cheese, were generally higher in the control group This result is due to limiting the micro organism reproduction by limiting the gas passage of the coatings. Hardness and chewiness values of the textural properties of the samples were significantly reduced in uncoated control samples compared to the coated samples due to maturation. The chitosan film coatings used in the study limited the development of mold yeast until the 30th day but after that did not yield successful results in this respect.

Keywords: chitosan, edible film, transglutaminase, whey protein

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Authors: Nadine khadraoui, Rym Essid, Olfa Tabbene, Imen Chniba, Safa Boujemaa, Selim Jallouli, Nadia Fares, Behija Mlik, Boutheina Ben Abdelmoumen Mardassi

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Background and aim: Mycoplasma hominis is an opportunistic pathogen that can cause various gynecological infections such cervicitis, infertility, and, less frequently, extra-genital infections. Previous studies on the antimicrobial susceptibility of Mycoplasma hominis Tunisian strains have highlighted a significant resistance, even multi-resistance, to the most used antibiotic in the therapy of consequential infections. To address this concern, the present study aimed for the alternative of phytotherapy. Peganum harmala seed extract was tested as an antibacterial agent against multidrug-resistant M.hominis clinical strains. Material and Methods: Peganum harmala plant was collected from Ain Sebaa, Tabarka, North West region of Tunisia in April 2018, air-dried, grounded and extracted by different solvents.The crude methanolic extract was further partitioned with n-HEX, DCM, EtOAC and n-BuOl. Antibacterial activity was evaluated against M. hominis ATCC 23114 and 20 M. hominis clinical strains.The antimycoplasmal activity was tested by the microdilution method, and MIC values were determined. Phytochemical analysis and hemolytic activity on human erythrocytes were also performed. The active fraction was then subjected to purification, and the chemical identification of the active compound was investigated. Results: Among the tested fractions, the n-BuOH extract was the most active fraction since it exhibited an inhibitory effect against M. hominis ATCC 23114 and 80% of the tested clinical strains with MIC between 125 and 1000 µg/ml. The phytochemical analysis of the n-BuOH revealed its metabolic abundance in polyphenols, flavonoids and condensed tannin with levels of 257.37 mg AGE/g, 172.27 mg EC/g and 58.27 mg EC/g, respectively. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against all M. hominis isolates with CMB values ranging between 125 and 4000 µg/ml. Further, the active fraction exhibited weak cytotoxicity effect at active concentrations when tested on human erythrocytes. The active compound was identified by gas chromatography–mass spectrometry as an indole alkaloid harmaline. Conclusion: In summary, Peganum harmala extract demonstrated an interesting anti-mycoplasmal activity against M. hominis Tunisian strains. Therefore, it could be considered as a potential candidate for the treatment of consequential infections. However, further studies are necessary to evaluate its mechanism of action in mycoplasmas.

Keywords: mycoplasma hominis, peganum harmala, antibioresistance, phytotherapy, phytochemical analysis

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Authors: Siddharth Deshpande, Nihar Masurkar, Vallerinteavide Mavelli Girish, Malan Desai, Chester Drum

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Folding and stabilization of recombinant proteins remain a consistent challenge for industrial and therapeutic applications. Proteins derived from thermophilic bacteria often have superior expression and stability qualities. To develop a generalizable approach to protein folding and stabilization, we tested the hypothesis that wrapping a thermostable exoshell around a protein substrate would aid folding and impart thermostable qualities to the internalized substrate. To test the effect of internalizing a protein within a thermostable exoshell (tES), we tested in vitro folding and stability using green fluorescent protein (GFPuv), horseradish peroxidase (HRP) and renilla luciferase (rLuc). The 8nm interior volume of a thermostable ferritin assembly was engineered to accommodate foreign proteins and either present a positive, neutral or negative interior charge environment. We further engineered the tES complex to reversibly assemble and disassemble with pH titration. Template proteins were expressed as inclusion bodies and an in vitro folding protocol was developed that forced proteins to fold inside a single tES. Functional yield was improved 100-fold, 100-fold and 150-fold with use of tES for GFPuv, HRP and rLuc respectively and was highly dependent on the internal charge environment of the tES. After folding, functional proteins could be released from the tES folding cavity using size exclusion chromatography at pH 5.8. Internalized proteins were tested for improved stability against thermal, organic, urea and guanidine denaturation. Our results demonstrated that thermostable exoshells can efficiently refold and stabilize inactive aggregates into functional proteins.

Keywords: thermostable shell, in vitro folding, stability, functional yield

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Authors: Lei Wang, Yingzhou Ni, Amr M. Bakry, Hao Cheng, Li Liang

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Food proteins have been widely used as carrier materials because of their multiple functional properties. In this study, alfa-tocopherol was encapsulated in the oil phase of an oil-in-water emulsion stabilized with whey protein isolate (WPI). The influence of WPI concentration and resveratrol or ascorbic acid on the decomposition of alfa-tocopherol in the emulsion during storage is discussed. Decomposition decreased as WPI concentrations increased. Decomposition was delayed at ascorbic acid/WPI molar ratios lower than 5 but was promoted at higher ratios. Resveratrol partitioned into the oil-water interface by binding to WPI and its cis-isomer is believed to have contributed most of the protective effect of this polyphenol. These results suggest the possibility of using the emulsifying and ligand-binging properties of WPI to produce carriers for simultaneous encapsulation of alfa-tocopherol and resveratrol in a single emulsion system.

Keywords: stability, alfa-tocopherol, resveratrol, whey protein isolate

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Authors: Akanksha Agrawal, Richa Rathor, Geetha Suryakumar

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Muscle represents about ¾ of the body mass, and a healthy muscular system is required for human performance. A healthy muscular system is dynamically balanced via the catabolic and anabolic process. High altitude associated hypoxia altered this redox balance via producing reactive oxygen and nitrogen species that ultimately modulates protein structure and function, hence, disrupts proteostasis or protein homeostasis. The mechanism by which proteostasis is clinched includes regulated protein translation, protein folding, and protein degradation machinery. Perturbation in any of these mechanisms could increase proteome imbalance in the cellular processes. Altered proteostasis in skeletal muscle is likely to be responsible for contributing muscular atrophy in response to hypoxia. Therefore, we planned to elucidate the mechanism involving altered proteostasis leading to skeletal muscle atrophy under chronic hypobaric hypoxia. Material and Methods-Male Sprague Dawley rats weighing about 200-220 were divided into five groups - Control (Normoxic animals), 1d, 3d, 7d and 14d hypobaric hypoxia exposed animals. The animals were exposed to simulated hypoxia equivalent to 282 torr pressure (equivalent to an altitude of 7620m, 8% oxygen) at 25°C. On completion of chronic hypobaric hypoxia (CHH) exposure, rats were sacrificed, muscle was excised and biochemical, histopathological and protein synthesis signaling were studied. Results-A number of changes were observed with the CHH exposure time period. ROS was increased significantly on 07 and 14 days which were attributed to protein oxidation via damaging muscle protein structure by oxidation of amino acids moiety. The oxidative damage to the protein further enhanced the various protein degradation pathways. Calcium activated cysteine proteases and other intracellular proteases participate in protein turnover in muscles. Therefore, we analysed calpain and 20S proteosome activity which were noticeably increased at CHH exposure as compared to control group representing enhanced muscle protein catabolism. Since inflammatory markers (myokines) affect protein synthesis and triggers degradation machinery. So, we determined inflammatory pathway regulated under hypoxic environment. Other striking finding of the study was upregulation of Akt/PKB translational machinery that was increased on CHH exposure. Akt, p-Akt, p70 S6kinase, and GSK- 3β expression were upregulated till 7d of CHH exposure. Apoptosis related markers, caspase-3, caspase-9 and annexin V was also increased on CHH exposure. Conclusion: The present study provides evidence of disrupted proteostasis under chronic hypobaric hypoxia. A profound loss of muscle mass is accompanied by the muscle damage leading to apoptosis and cell death under CHH. These cellular stress response pathways may play a pivotal role in hypobaric hypoxia induced skeletal muscle atrophy. Further research in these signaling pathways will lead to development of therapeutic interventions for amelioration of hypoxia induced muscle atrophy.

Keywords: Akt/PKB translational machinery, chronic hypobaric hypoxia, muscle atrophy, protein degradation

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Authors: Samashi Munaweera

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Cancer stem cells (CSC) are responsible for the initiation, extensive proliferation and metastasis of cancer. CSCs, including breast cancer stem cells (bCSCs) have a capacity to generate chemo and radiotherapy resistance heterogeneous population of cells. Over-expressed ABCB1 has been reported as a main reason for drug resistance of CSCs via activating drug efflux pumps by creating pores in the cell membrane. The overall efficiency of chemotherapeutic agents might be enhanced by blocking the ABCB protein efflux pump in the CSC membrane. There is an urgent need to search for persuasive natural drugs which can target CSCs. Anti-cancer properties of Hylocereus undatus on cancer CSCs have not yet been studied. In the present study, the anti-cancer effects of the peel and flesh of H. undatus fruit on bCSCs were evaluated with the aim of developing a marketable anti-cancer nutraceutical formulation. The flesh and peel of H. undatus were freeze-dried and sequentially extracted into four different solvents (hexane, chloroform, ethyl acetate and ethanol). All extracts (eight extracts) were dried under reduced pressure, and different concentrations (12.5-400 µg/mL) were treated on bCSCs isolated from a triple-negative chemo-resistant breast cancer phenotype (MDA-MB-231 cells). Anti-proliferative effects of all extracts and paclitaxel (positive control) were determined by a colorimetric assay (WST-1 based). Since peel-chloroform (IC50= 54.8 µg/mL) and flesh-ethyl acetate (IC50= 150.5 µg/mL) extras exerted a potent anti-proliferative effect at 72 h post-incubation, a combinatorial formulation (CF) was developed with the most active peel-chloroform extract and 20 µg/mL of verapamil (a known ABCB1 drug efflux pump blocker) first time in the world. Anti-proliferative effects and pro-apoptotic effects of CF were confirmed by estimating activated caspase3 and caspase7 levels and apoptotic morphological features in the CF-treated bCSCs compared to untreated and only verapamil (20 µg/mL) treated bCSCs, and CF treated normal mammary epithelial cells (MCF-10A). The antiproliferative effects of CF (16.4 µg/mL) are greater than paclitaxel (19.2 µg/mL) and three folds greater than peel-chloroform extract (IC50= 54.8 µg/mL) on bCSCs while exerting less effects on normal cells (> 400 µg/mL). Collectively, CF can be considered as a potential initiative of a nutraceutical formulation that can target CSCs.

Keywords: breast cancer stem cells (bCSCs), Hylocereus undatus, combinatorial formulation (CF), ABCB 1 protein, verapamil

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Authors: Demetrio L. Valle Jr., Juliana Janet M. Puzon, Windell L. Rivera

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From the various medicinal plants available in the Philippines, crude ethanol extracts of twelve (12) Philippine medicinal plants, namely: Senna alata L. Roxb. (akapulko), Psidium guajava L. (bayabas), Piper betle L. (ikmo), Vitex negundo L. (lagundi), Mitrephora lanotan (Blanco) Merr. (Lanotan), Zingiber officinale Roscoe (luya), Curcuma longa L. (Luyang dilaw), Tinospora rumphii Boerl (Makabuhay), Moringga oleifera Lam. (malunggay), Phyllanthus niruri L. (sampa-sampalukan), Centella asiatica (L.) Urban (takip kuhol), and Carmona retusa (Vahl) Masam (tsaang gubat) were studied. In vitro methods of evaluation against selected Gram-positive and Gram-negative multidrug-resistant (MDR), bacteria were performed on the plant extracts. Although five of the plants showed varying antagonistic activities against the test organisms, only Piper betle L. exhibited significant activities against both Gram-negative and Gram-positive multidrug-resistant bacteria, exhibiting wide zones of growth inhibition in the disk diffusion assay, and with the lowest concentrations of the extract required to inhibit the growth of the bacteria, as supported by the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays. Further antibacterial studies of the Piper betle L. leaf, obtained by three extraction methods (ethanol, methanol, supercritical CO2), revealed similar inhibitory activities against a multitude of Gram-positive and Gram-negative MDR bacteria. Thin layer chromatography (TLC) assay of the leaf extract revealed a maximum of eight compounds with Rf values of 0.92, 0.86, 0.76, 0.53, 0.40, 0.25, 0.13, and 0.013, best visualized when inspected under UV-366 nm. TLC- agar overlay bioautography of the isolated compounds showed the compounds with Rf values of 0.86 and 0.13 having inhibitory activities against Gram-positive MDR bacteria (MRSA and VRE). The compound with an Rf value of 0.86 also possesses inhibitory activity against Gram-negative MDR bacteria (CRE Klebsiella pneumoniae and MBL Acinetobacter baumannii). Gas Chromatography-Mass Spectrometry (GC-MS) was able to identify six volatile compounds, four of which are new compounds that have not been mentioned in the medical literature. The chemical compounds isolated include 4-(2-propenyl)phenol and eugenol; and the new four compounds were ethyl diazoacetate, tris(trifluoromethyl)phosphine, heptafluorobutyrate, and 3-fluoro-2-propynenitrite. Phytochemical screening and investigation of its antioxidant, cytotoxic, possible hemolytic activities, and mechanisms of antibacterial activity were also done. The results showed that the local variant of Piper betle leaf extract possesses significant antioxidant, anti-cancer and antimicrobial properties, attributed to the presence of bioactive compounds, particularly of flavonoids (condensed tannin, leucoanthocyanin, gamma benzopyrone), anthraquinones, steroids/triterpenes and 2-deoxysugars. Piper betle L. is also traditionally known to enhance wound healing, which could be primarily due to its antioxidant, anti-inflammatory and antimicrobial activities. In vivo studies on mice using 2.5% and 5% of the ethanol leaf extract cream formulations in the excised wound models significantly increased the process of wound healing in the mice subjects, the results and values of which are at par with the current antibacterial cream (Mupirocin). From the results of the series of studies, we have definitely proven the value of Piper betle L. as a source of bioactive compounds that could be developed into therapeutic agents against MDR bacteria.

Keywords: Philippine herbal medicine, multidrug-resistant bacteria, Piper betle, TLC-bioautography

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Authors: Iftikhar Tayubi, Tabrej Khan, Rayan Alsulmi, Abdulrahman Labban

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Spider produces a special kind of substance. This special kind of substance is called a toxin. The toxin is composed of many types of protein, which differs from species to species. Spider toxin consists of several proteins and non-proteins that include various categories of toxins like myotoxin, neurotoxin, cardiotoxin, dendrotoxin, haemorrhagins, and fibrinolytic enzyme. Protein Sequence information with references of toxins was derived from literature and public databases. From the previous findings, the Spider toxin would be the best choice to treat different types of tumors and cancer. There are many therapeutic regimes, which causes more side effects than treatment hence a different approach must be adopted for the treatment of cancer. The combinations of drugs are being encouraged, and dramatic outcomes are reported. Spider toxin is one of the natural cytotoxic compounds. Hence, it is being used to treat different types of tumors; especially its positive effect on breast cancer is being reported during the last few decades. The efficacy of this database is that it can provide a user-friendly interface for users to retrieve the information about Spiders, toxin and toxin protein of different Spiders species. SPIDTOXD provides a single source information about spider toxins, which will be useful for pharmacologists, neuroscientists, toxicologists, medicinal chemists. The well-ordered and accessible web interface allows users to explore the detail information of Spider and toxin proteins. It includes common name, scientific name, entry id, entry name, protein name and length of the protein sequence. The utility of this database is that it can provide a user-friendly interface for users to retrieve the information about Spider, toxin and toxin protein of different Spider species. The database interfaces will satisfy the demands of the scientific community by providing in-depth knowledge about Spider and its toxin. We have adopted the methodology by using A MySQL and PHP and for designing, we used the Smart Draw. The users can thus navigate from one section to another, depending on the field of interest of the user. This database contains a wealth of information on species, toxins, and clinical data, etc. This database will be useful for the scientific community, basic researchers and those interested in potential pharmaceutical Industry.

Keywords: siptoxd, php, mysql, toxin

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Authors: H.Yari, M. Mohseni, Z. Ranjbar

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In this work a typical automotive clearcoat is modified with a combination of hyperbranched polymer (HBP) and polyhedral oligomeric silsesquioxane (POSS) nanostructures to simultaneously enhance the scratch resistance and healing ability of the resulting films. Micro-scratch and healing data revealed that these goals were achieved at high loadings of modifiers. Enhanced scratch resistance was attributed to the improved elastic recovery of the clearcoats in presence of modifiers. In addition, improved healing performance due to the partial replacement of covalent cross-links with physical ones resulted from the unique globular highly branched structure of HBP and POSS macromolecules.

Keywords: automotive clearcoat, POSS building blocks scratch resistance, self-healing

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Authors: Ghibeche Abderrahmane

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To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n=7) and age-matched (n=5) and young (n=9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50,P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Keywords: euglycemic-hyperinsulinemic, HSP72, mRNA, diabete

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Authors: Adnan I. O. Zaid, Safwan M. A. Alqawabah

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Review of the available literature on grain refinement of aluminum and its alloys reveals that little work is published on the effect of refiners on mechanical characteristics and wear resistance. In this paper, the effect of addition of Zr to Al grain refined by Ti+B on its metallurgical, mechanical characteristics and wear resistance both in the as cast and after extrusion condition are presented and discussed. It was found that Addition of Zr to Al resulted in deterioration of its mechanical strength and hardness, whereas it resulted in improvement of both of them when added to Al grain refined by Ti+B. Furthermore it was found that the direct extrusion process resulted in further increase of the mechanical strength and hardness of Al and its micro-alloys. Also it resulted in increase of their work hardening index, n, i.e. improved their formability, hence it reduces the number of stages required for forming at large strains in excess of the plastic instability before Zr addition.

Keywords: aluminum, grain refinement, titanium + boron, zirconium, mechanical characteristics, wear resistance, direct extrusion

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Authors: Irma Gabriela Lopez-Moreno, Elva Perez-Luque, Herlinda Aguilar-Zavala

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Introduction: Type 2 Diabetes Mellitus (T2DM) is characterized by combination of insulin resistance and deterioration of insulin secretion. Sfrp5 is a protein that antagonizes Wnt5a proteins by preventing it from reaching its receptor and activating the Wnt/β-catenin signaling pathway, this pathway is one of the most important regulators of adipogenesis. Although metformin decreases glucose levels its mechanisms of action are not fully known but it has been implicated in the inhibition of the Wnt/β-catenin signaling pathway. Objective: The objective was evaluating the effects of metformin on serum levels of Sfrp5 in patients with T2DM treated and not treated with metformin. Methods: Two groups of patients were selected: one group of T2DM patients treated with metformin (n = 35) and another group of subjects with recent diagnosis of T2DM untreated (n = 35) with a mean age of 48 ± 9 years. In these subjects anthropometric measures were taken as weight, height, waist and hip circumference, were calculated the percentage of body fat, visceral fat and muscle mass. In addition, were measured glucose levels, lipid profile, adiponectin and Sfrp5. Results: Sfrp5 were higher in metformin-treated patients compared to the untreated group (19.9 vs 13.6 ng/mL p < 0.001), a negative correlation was found between Sfrp5 levels and total cholesterol levels (r= -0.25, p = 0.03) and percentage of visceral fat (r = -0.26, p = 0.03) and a positive correlation with HDL cholesterol levels (r = 0.31, p = 0.01) and adiponectin (r=0.65, p = < 0.001). Conclusions: The findings show that metformin consumption increased levels of Sfrp5, which may lead to a decrease in the activation of the WNT/β-catenin pathway impacting on adipogenesis.

Keywords: adiponectin, diabetes, metformin, Sfrp5

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Authors: Marvin R. Tenecio

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The analysis of "religious resistance" has been prevalent in Asian and Philippine studies. Discussions on religious resistance from a variety of perspectives are deemed as crucial turning points in the concept's ongoing development and expansion. By broadening the backdrop of religious protest between the early 1960s and the late 1980s, the researchers contend that a study examining the role carried by the Catholic Church in the upheavals against dictatorships in South Korea and the Philippines would be beneficial to the body of knowledge. This study examines a variety of historical writings about the activities occurring at that time. The researchers also compare and contrast the Catholic Church's contributions to the Korean and Philippine resistance against Park Chung-Hee and Ferdinand Marcos Sr., respectively, during the early 1960s until the late 1980s, using the lens of history from below, particularly the Pasyon and Revolution. The Catholic Church stood out against human rights abuses, promoted social justice, and mobilized the public for political reform in response to the dictatorships in South Korea and the Philippines. Even though the specific circumstances and personalities may have changed, the Church's position in both countries was vital in opposing authoritarian governments and supporting democratic movements.

Keywords: resistance, movements, catholic, church, dictatorship

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Authors: Sihame Amakrane, Zineb Ouahdi, Mohammed Salah, Said Belaaouad

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\This research explores the efficacy of novel pyrimidine derivatives on bacterial strains such as Escherichia coli, Staphylococcus aureus, and Myccobacterium tuberculosis, utilizing bending energy calculations. Of the 25 compounds examined, 13 displayed potent activity against all the bacterial strains under study, exhibiting bending energy measurements between -7.4 and -10.7 kcal/mol. The -7.4 kcal/mol value corresponds to the bending energy of the SA12 and SA13 compounds with the 2xct protein (Staphylococcus aureus), whereas the -10.7 kcal/molis linked with the bending energy of SA6 and SA11 compounds with the 6GAV protein (Myccobacterium tuberculosis). Further research will involve a QSAR (Quantitative Structure-Activity Relationship) study aimed at constructing a reliable model to combat the aforementioned bacterial strains and a molecular dynamics study to evaluate the stability of ligand-protein complexes.

Keywords: docking, QSAR, bending energy, e. coli

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Authors: Mayank, Navneet Kaur, Narinder Singh

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The V599 mutations in the BRAF protein are extremely oncogenic, responsible for countless of malignant conditions. Along with wild type, V599E, V599D, and V599R are the important mutated variants of the BRAF proteins. The BRAF inhibitory anticancer agents are continuously developing, and sorafenib is a BRAF inhibitor that is under clinical use. The crystal structure of sorafenib bounded to wild type, and V599 is known, showing a similar interaction pattern in both the case. The mutated 599th residue, in both the case, is also found not interacting directly with the co-crystallized sorafenib molecule. However, the IC50 value of sorafenib was found extremely different in both the case, i.e., 22 nmol/L for wild and 38 nmol/L for V599E protein. Molecular docking study and MMGBSA binding energy results also revealed a significant difference in the binding pattern of sorafenib in both the case. Therefore, to explore the role of distinctively situated 599th residue, we have further conducted comprehensive computational studies. The molecular dynamics simulation, residue interaction network (RIN) analysis, and residue correlation study results revealed the importance of the 599th residue on the therapeutic outcome and overall dynamic of the BRAF protein. Therefore, although the position of 599th residue is very much distinctive from the ligand-binding cavity of BRAF, still it has exceptional control over the overall functional outcome of the protein. The insight obtained here may seem extremely important and guide us while designing ideal BRAF inhibitory anticancer molecules.

Keywords: BRAF, oncogenic, sorafenib, computational studies

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Authors: Mohammad Mardi

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Generally, the resistivity is high in oil layer and low in water layer. Yet there are intervals of oil-bearing zones showing low resistivity, high porosity, and low resistance. In the typical example, well A (depth: 4341.5-4372.0m), both Spectral Gamma Ray (SGR) and Corrected Gamma Ray (CGR) are relatively low; porosity varies from 12-22%. Above 4360 meters, the reservoir shows the conventional positive difference between deep and shallow resistivity with high resistance; below 4360m, the reservoir shows a negative difference with low resistance, especially at depths of 4362.4 meters and 4371 meters, deep resistivity is only 2Ω.m, and the CAST-V imaging map shows that there are low resistance substances contained in the pores or matrix in the reservoirs of this interval. The rock slice analysis data shows that the pyrite volume is 2-3% in the interval 4369.08m-4371.55m. A comprehensive analysis on the volume of shale (Vsh), porosity, invasion features of resistivity, mud logging, and mineral volume indicates that the possible causes for the negative difference between deep and shallow resistivities with relatively low resistance are erosional pores, caves, micritic texture and the presence of pyrite. Full-bore Drill Stem Test (DST) verified 4991.09 bbl/d in this interval. To identify and thoroughly characterize low resistivity intervals coring, Nuclear Magnetic Resonance (NMR) logging and further geological evaluation are needed.

Keywords: low resistivity pay, carbonates petrophysics, microporosity, porosity

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Authors: Olubukayo-Opeyemi Oyetayo, Oscar Mendez-Lucio, Andreas Bender, Hans Kiefer

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The thermal melting curve of a protein provides information on its conformational stability and could provide cues on its aggregation behavior. Naturally-occurring osmolytes have been shown to improve the thermal stability of most proteins in a concentration-dependent manner. They are therefore commonly employed as additives in therapeutic protein purification and formulation. A number of intertwined and seemingly conflicting mechanisms have been put forward to explain the observed stabilizing effects, the most prominent being the preferential exclusion mechanism. We attempted to probe and summarize molecular mechanisms for thermal stabilization of a monoclonal antibody (mAb) by developing quantitative structure-activity relationships using a rationally-selected library of 120 osmolyte-like compounds in the polyhydric alcohols, amino acids and methylamines classes. Thermal stabilization potencies were experimentally determined by thermal shift assays based on differential scanning fluorimetry. The cross-validated QSAR model was developed by partial least squares regression using descriptors generated from Molecular Operating Environment software. Careful evaluation of the results with the use of variable importance in projection parameter (VIP) and regression coefficients guided the selection of the most relevant descriptors influencing mAb thermal stability. For the mAb studied and at pH 7, the thermal stabilization effects of tested compounds correlated positively with their fractional polar surface area and inversely with their fractional hydrophobic surface area. We cannot claim that the observed trends are universal for osmolyte-protein interactions because of protein-specific effects, however this approach should guide the quick selection of (de)stabilizing compounds for a protein from a chemical library. Further work with a large variety of proteins and at different pH values would help the derivation of a solid explanation as to the nature of favorable osmolyte-protein interactions for improved thermal stability. This approach may be beneficial in the design of novel protein stabilizers with optimal property values, especially when the influence of solution conditions like the pH and buffer species and the protein properties are factored in.

Keywords: thermal stability, monoclonal antibodies, quantitative structure-activity relationships, osmolytes

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Authors: Narin Salehiyan, Ramin Ghasemi Shayan

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In order to investigate coronavirus RNA replication, transcription, recombination, protein processing and transport, virion assembly, the identification of coronavirus-specific cell receptors, and polymerase processing, the manipulation of coronavirus clones and complementary DNAs (cDNAs) of defective-interfering (DI) RNAs is the subject of this chapter. The idea of the Covid genome is nonsegmented, single-abandoned, and positive-sense RNA. When compared to other RNA viruses, its size is significantly greater, ranging from 27 to 32 kb. The quality encoding the enormous surface glycoprotein depends on 4.4 kb, encoding a forcing trimeric, profoundly glycosylated protein. This takes off exactly 20 nm over the virion envelope, giving the infection the appearance-with a little creative mind of a crown or coronet. Covid research has added to the comprehension of numerous parts of atomic science as a general rule, like the component of RNA union, translational control, and protein transport and handling. It stays a fortune equipped for creating startling experiences.

Keywords: covid-19, corona, virus, genome, genetic

Procedia PDF Downloads 65

Authors: Awf A. Al-Khan, Michael J. Day, Judith Nimmo, Mourad Tayebi, Stewart D. Ryan, Samantha J. Richardson, Janine A. Danks

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Osteosarcoma (OS) is the most common type of malignant primary bone tumour in dogs. In addition to their critical roles in bone formation and remodeling, parathyroid hormone-related protein (PTHrP) and its receptor (PTHR1) are involved in progression and metastasis of many types of tumours in humans. The aims of this study were to determine the localisation and expression levels of PTHrP and PTHR1 in canine OS tissues using immunohistochemistry and to investigate if this expression is correlated with survival time. Formalin-fixed, paraffin-embedded tissue samples from 44 dogs with known survival time that had been diagnosed with primary osteosarcoma were analysed for localisation of PTHrP and PTHR1. Findings showed that both PTHrP and PTHR1 were present in all OS samples. The dogs with high level of PTHR1 protein (16%) had decreased survival time (P<0.05) compared to dogs with less PTHR1 protein. PTHrP levels did not correlate with survival time (P>0.05). The results of this study indicate that the PTHR1 is expressed differently in canine OS tissues and this may be correlated with poor prognosis. This may mean that PTHR1 may be useful as a prognostic indicator in canine OS and could represent a good therapeutic target in OS.

Keywords: dog, expression, osteosarcoma, parathyroid hormone receptor 1 (PTHR1), parathyroid hormone-related protein (PTHrP), survival

Procedia PDF Downloads 267

Authors: Claudia Cabezas-Zabala, Jairo Lindarte-Artunduaga, Carlos Mario Zuluaga-Dominguez

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Amaranth is an emerging pseudocereal rich in such essential nutrients as protein and dietary fiber, which was employed as an ingredient in the formulation of crackers to evaluate the rheological performance and sensory acceptability of the obtained food. A completely randomized factorial design was used with two factors: (A) ratio of wheat and amaranth flour used in the preparation of the dough, in proportion 90:10 and 80:20 (% w/w) and (B) two levels of inulin addition of 8.4% and 16.7 %, having two control doughs made from amaranth and wheat flour, respectively. Initially, the functional properties of the formulations mentioned were measured, showing no significant differences in the water absorption capacity (WAC) and swelling power (SP), having mean values between 1.66 and 1.81 g/g for WAC and between 1.75 and 1.86 g/g for SP, respectively. The amaranth flour had the highest water holding capacity (WHR) of 8.41 ± 0.15 g/g and emulsifying activity (EA) of 74.63 ± 1.89 g/g. Moreover, the rheological behavior, measured through the use of farinograph, extensograph, Mixolab, and falling index, showed that the formulation containing 20% of amaranth flour and 7.16% of inulin had a rheological behavior similar to the control produced exclusively with wheat flour, being the former, the one selected for the preparation of crackers. For this formulation, the farinograph showed a mixing tolerance index of 11 UB, indicating a strong and cohesive dough; likewise, the Mixolab showed dough reaches stability at 6.47 min, indicating a good resistance to mixing. On the other hand, the extensograph exhibited a dough resistance of 637 UB, as well as extensibility of 13.4 mm, which corresponds to a strong dough capable of resisting the laminate. Finally, the falling index was 318 s, which indicates the crumb will retain enough air to enhance the crispness of a characteristic cracker. Finally, a sensory consumer test did not show significant differences in the evaluation of aroma between the control and the selected formulation, while this latter had a significantly lower rating in flavor. However, a purchase intention of 70 % was observed among the population surveyed. The results obtained in this work give perspectives for the industrial use of amaranth in baked goods. Additionally, amaranth has been a product typically linked to indigenous populations in the Andean South American countries; therefore, the search for diversification and alternatives of use for this pseudocereal has an impact on the social and economic conditions of such communities. The technological versatility and nutritional quality of amaranth is an advantage for consumers, favoring the consumption of healthy products with important contributions of dietary fiber and protein.

Keywords: amaranth, crackers, rheology, pseudocereals, kneaded products

Procedia PDF Downloads 115

Authors: Chuanpeng Zhou

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The effect of dietary carbohydrate (CHO) level on serum physiological response, hepatic antioxidant abilities and heat shock protein 70 (HSP70) expression of Wuchang bream (Megalobrama amblycephala) was studied. Two isonitrogenous (28.56% crude protein) and isolipidic (5.28% crude lipid) diets were formulated to contain 30% or 53% wheat starch. Diets were fed for 90 days to fish in triplicate tanks (28 fish per tank). At the end of feeding trial, significantly higher serum triglyceride level, insulin level, cortisol level, malondialdehyde (MDA) content were observed in fish fed the 53% CHO diet, while significantly lower serum total protein content, alkaline phosphatase (AKP) activity, superoxide dismutase (SOD) activity and total antioxidative capacity (T-AOC) were found in fish fed the 53% CHO diet compared with those fed the 30% diet. The relative level of hepatic heat shock protein 70 mRNA was significantly higher in the 53% CHO group than that in the 30% CHO at 6, 12, and 48 h after feeding. The results of this study indicated that ingestion of 53% dietary CHO impacted the nonspecific immune ability and caused metabolic stress of Megalobrama amblycephala.

Keywords: Megalobrama amblycephala, carbohydrate, fasted and postprandial response, immunity, Hsp70

Procedia PDF Downloads 453

Authors: Coulibaly Fatoumata, Seme Kpassi, Aboly Nicolas, Ccoulibaly Zonzereke

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Introduction and Objective: Microbial resistance is nowadays a major public health problem. In the perspective of a better understanding of the resistance of ticks against acaricides, a study was conducted in the Department of Korhogo. The general objective was to verify the knowledge and skills of breeders on the use of acaricides and contribute to reducing the impact of ticks on livestock productivity. Methodology: To carry out the work, a cross-sectional survey was conducted using elaborate questionnaires, followed by specific interviews with livestock stakeholders in the Korhogo sub-prefecture. Results: The results showed that in the study area, cattle breeders, the majority of whom (58.06%) are Ivorians, use anti-ticks without strict compliance with recommendations of the instructions and standards for use. 68% of them performed under-dosed treatments, and 32% an over-dosed treatment. The most common method for treating cattle against ticks was spraying. Conclusion: Despite the use of tick repellents by these breeders, tick-borne diseases still persist. This could be explained by the misuse of the products (under dosage and overdose), which can cause harmful effects or even resistance of certain ticks. It is, therefore important to respect the normal dosage of the products used as well as the methods of use (bath, spray, pour-on, etc.). In order to minimize the problems of resistance, awareness is necessary among breeders for the proper use of acaricidal products as well as all other drugs.

Keywords: ticks, resistance, anti-tick, cattle, korhogo

Procedia PDF Downloads 74

Authors: A. Ameur Ameur, F. Chougrani, M. Halbouche

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In order to characterize the sheep's milk, we analyzed and compared, in a first stage of our work, the physical and chemical characteristics in two Algerian sheep breeds: Hamra race and race Ouled Djellal breeding at the station the experimental ITELV Ain Hadjar (Saïda Province). Analyses are performed by Ekomilk Ultra-analyzer (EON TRADING LLC, USA), they focused on the pH, density, freezing, fat, total protein, solids-the total dry extract. The results obtained for these parameters showed no significant differences between the two breeds studied. The second stage of this work was the isolation and characterization of milk proteins. For this, we used the precipitation of caseins phi [pH 4.6]. For this, we used the precipitation of caseins Phi (pH 4.6). After extraction, purification and assay, both casein and serum protein fractions were then assayed by the Bradford method and controlled by polyacrylamide gel electrophoresis (PAGE) in the different conditions (native, in the presence of urea and in the presence of SDS). The electrophoretic pattern of milk samples showed the presence similarities of four major caseins variants (αs1-, αs2-β-and k-casein) and two whey proteins (β-lactoglobulin, α-lactalbumin) of two races Hamra and Ouled Djellal. But compared to bovine milk, they have helped to highlight some peculiarities as related to serum proteins (α La β Lg) as caseins, including αs1-Cn.

Keywords: Hamra, Ouled Djellal, protein polymorphism, sheep breeds

Procedia PDF Downloads 554

Authors: Annamária Käferné Rácz, Bence Jáger, Balázs Kövesdi, László Dunai

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Due to the numerous advantages of steel corrugated web girders, its application field is growing for bridges as well as for buildings. The global stability behavior of such girders is significantly larger than those of conventional I-girders with flat web, thus the application of the structural steel material can be significantly reduced. Design codes and specifications do not provide clear and complete rules or recommendations for the determination of the lateral torsional buckling (LTB) resistance of corrugated web girders. Therefore, the authors made a thorough investigation regarding the LTB resistance of the corrugated web girders. Finite element (FE) simulations have been performed to develop new design formulas for the determination of the LTB resistance of trapezoidally corrugated web girders. FE model is developed considering geometrical and material nonlinear analysis using equivalent geometric imperfections (GMNI analysis). The equivalent geometric imperfections involve the initial geometric imperfections and residual stresses coming from rolling, welding and flame cutting. Imperfection sensitivity analysis was performed to determine the necessary magnitudes regarding only the first eigenmodes shape imperfections. By the help of the validated FE model, an extended parametric study is carried out to investigate the LTB resistance for different trapezoidal corrugation profiles. First, the critical moment of a specific girder was calculated by FE model. The critical moments from the FE calculations are compared to the previous analytical calculation proposals. Then, nonlinear analysis was carried out to determine the ultimate resistance. Due to the numerical investigations, new proposals are developed for the determination of the LTB resistance of trapezoidally corrugated web girders through a modification factor on the design method related to the conventional flat web girders.

Keywords: corrugated web, lateral torsional buckling, critical moment, FE modeling

Procedia PDF Downloads 279

Authors: Hamidreza Saberkari, Mousa Shamsi, Hossein Ahmadi, Saeed Vaali, , MohammadHossein Sedaaghi

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In the recent years, using signal processing tools for accurate identification of the protein coding regions has become a challenge in bioinformatics. Most of the genomic signal processing methods is based on the period-3 characteristics of the nucleoids in DNA strands and consequently, spectral analysis is applied to the numerical sequences of DNA to find the location of periodical components. In this paper, a novel ensemble algorithm for gene selection in DNA sequences has been presented which is based on the combination of Goertzel algorithm and anti-notch filter (ANF). The proposed algorithm has many advantages when compared to other conventional methods. Firstly, it leads to identify the coding protein regions more accurate due to using the Goertzel algorithm which is tuned at the desired frequency. Secondly, faster detection time is achieved. The proposed algorithm is applied on several genes, including genes available in databases BG570 and HMR195 and their results are compared to other methods based on the nucleotide level evaluation criteria. Implementation results show the excellent performance of the proposed algorithm in identifying protein coding regions, specifically in identification of small-scale gene areas.

Keywords: protein coding regions, period-3, anti-notch filter, Goertzel algorithm

Procedia PDF Downloads 384