Search results for: phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin
4353 In silico Designing of Imidazo [4,5-b] Pyridine as a Probable Lead for Potent Decaprenyl Phosphoryl-β-D-Ribose 2′-Epimerase (DprE1) Inhibitors as Antitubercular Agents
Authors: Jineetkumar Gawad, Chandrakant Bonde
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Tuberculosis (TB) is a major worldwide concern whose control has been exacerbated by HIV, the rise of multidrug-resistance (MDR-TB) and extensively drug resistance (XDR-TB) strains of Mycobacterium tuberculosis. The interest for newer and faster acting antitubercular drugs are more remarkable than any time. To search potent compounds is need and challenge for researchers. Here, we tried to design lead for inhibition of Decaprenyl phosphoryl-β-D-ribose 2′-epimerase (DprE1) enzyme. Arabinose is an essential constituent of mycobacterial cell wall. DprE1 is a flavoenzyme that converts decaprenylphosphoryl-D-ribose into decaprenylphosphoryl-2-keto-ribose, which is intermediate in biosynthetic pathway of arabinose. Latter, DprE2 converts keto-ribose into decaprenylphosphoryl-D-arabinose. We had a selection of 23 compounds from azaindole series for computational study, and they were drawn using marvisketch. Ligands were prepared using Maestro molecular modeling interface, Schrodinger, v10.5. Common pharmacophore hypotheses were developed by applying dataset thresholds to yield active and inactive set of compounds. There were 326 hypotheses were developed. On the basis of survival score, ADRRR (Survival Score: 5.453) was selected. Selected pharmacophore hypotheses were subjected to virtual screening results into 1000 hits. Hits were prepared and docked with protein 4KW5 (oxydoreductase inhibitor) was downloaded in .pdb format from RCSB Protein Data Bank. Protein was prepared using protein preparation wizard. Protein was preprocessed, the workspace was analyzed using force field OPLS 2005. Glide grid was generated by picking single atom in molecule. Prepared ligands were docked with prepared protein 4KW5 using Glide docking. After docking, on the basis of glide score top-five compounds were selected, (5223, 5812, 0661, 0662, and 2945) and the glide docking score (-8.928, -8.534, -8.412, -8.411, -8.351) respectively. There were interactions of ligand and protein, specifically HIS 132, LYS 418, TRY 230, ASN 385. Pi-pi stacking was observed in few compounds with basic Imidazo [4,5-b] pyridine ring. We had basic azaindole ring in parent compounds, but after glide docking, we received compounds with Imidazo [4,5-b] pyridine as a basic ring. That might be the new lead in the process of drug discovery.Keywords: DprE1 inhibitors, in silico drug designing, imidazo [4, 5-b] pyridine, lead, tuberculosis
Procedia PDF Downloads 1544352 The Priming Effect of Morphology, Phonology, Semantics, and Orthography in Mandarin Chinese: A Prime Paradigm Study
Authors: Bingqing Xu, Wenxing Shuai
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This study investigates the priming effects of different Chinese compound words by native Mandarin speakers. There are lots of homonym, polysemy, and synonym in Chinese. However, it is unclear which kind of words have the biggest priming effect. Native Mandarin speakers were tested in a visual-word lexical decision experiment. The stimuli, which are all two-character compound words, consisted of two parts: primes and targets. Five types of relationships were used in all stimuli: morphologically related condition, in which the prime and the target contain the same morpheme; orthographically related condition, in which the target and the prime contain the different morpheme with the same form; phonologically related condition, in which the target and the prime contain the different morpheme with the same phonology; semantically related condition, in which the target and the prime contain the different morpheme with similar meanings; totally unrelated condition. The time since participants saw the target to respond was recorded. Analyses on reaction time showed that the average reaction time of morphologically related targets was much shorter than others, suggesting the morphological priming effect is the biggest. However, the reaction time of the phonologically related conditions was the longest, even longer than unrelated conditions. According to scatter plots analyses, 86.7% of participants had priming effects in morphologically related conditions, only 20% of participants had priming effects in phonologically related conditions. These results suggested that morphologically related conditions had the biggest priming effect. The orthographically and semantically related conditions also had priming effects, whereas the phonologically related conditions had few priming effects.Keywords: priming effect, morphology, phonology, semantics, orthography
Procedia PDF Downloads 1464351 Proximate and Mineral Composition of Chicken Giblets from Vojvodina, Northern Serbia
Authors: M. R. Jokanović, V. M. Tomović, M. T. Jović, S. B. Škaljac, B. V. Šojić, P. M. Ikonić, T. A. Tasić
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Proximate (moisture, protein, total fat, total ash) and mineral (K, P, Na, Mg, Ca, Zn, Fe, Cu and Mn) composition of chicken giblets (heart, liver and gizzard) were investigated. Phosphorous content, as well as proximate composition, were determined according to recommended ISO methods. The content of all elements, except phosphorus, of the giblets tissues were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES), after dry ashing mineralization. Regarding proximate composition heart was the highest in total fat content, and the lowest in protein content. Liver was the highest in protein and total ash content, while gizzard was the highest in moisture and the lowest in total fat content. Regarding mineral composition liver was the highest for K, P, Ca, Mg, Fe, Zn, Cu, and Mn, while heart was the highest for Na content. The contents of almost all investigated minerals in analysed giblets tissues of chickens from Vojvodina were similar to values reported in the literature, i.e. in national food composition databases of other countries.Keywords: chicken giblets, proximate composition, mineral composition, inductively coupled plasma-optical emission spectrometry (ICP-OES)
Procedia PDF Downloads 4514350 The Use of Caricatures as a Means of Advertising: The Case of Sütaş
Authors: Güldane Zengin
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Advertisers often make use of humorous elements in advertisements they create. Advertisements that contain such elements play a great role in creating awareness and attaining positive attitudes. Caricature, which is an element of humour, is interesting, eye catching, entertaining and memorable by its very nature. Because of these characteristics of caricatures, they are being used for advertising purposes. Advertisements with caricatures are spreading rapidly and embraced by consumers easily. Especially in the last ten years, companies in different sectors use advertisements with caricatures to publicize their products and services. These companies have different target audiences with different characteristics. They all have differences in opinions, attitudes, perceptions and buying behaviours. Target audiences’ brand choices depend on many different factors. Advertising is an important factor in brand choice. Using attention grabbing methods like advertising with caricatures affects their buying behaviours. This study examines the use of caricatures in Sütaş advertisements. Target audiences’ opinions, perceptions and attitudes about advertisements with caricatures are examined in this descriptive study.Keywords: advertising, advertisements with caricatures, caricature, communication, humour, Sütaş caricatures
Procedia PDF Downloads 2544349 Ring FingerPortein 2 (RNF2) Targeting by miRNAs in Breast Cancer Cell Lines
Authors: Ceyda Okudu, Secil Eroglu, Khandakar A. S. M. Saadat, Sibel O. Balci
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Ring Finger Protein 2 (RNF2) is a member of polycomb repressive complex 1 (PRC1), which is one of the epigenetic regulators in the genome. When RNF2 combines with other PRC1 members, it mediates the mono-ubiquitination of Histon2A (H2A). In breast cancer, RNF2 is commonly overexpressed, and also it promotes metastasis and invasion in other aggressive tumors like melanoma, prostate, and hepatocarcinoma. The role of RNF2 in the metastasis and invasion of breast cancer has not yet been elucidated. Our aim is to observe the role of RNF2 in metastasis and invasion in this study by miRNA mediated RNF2 gene silencing in breast cancer cell lines. We selected miRNAs, targeting to RNF2 by searching online databases. miR-17-5p, miR20a-5p, and miR-106b-5p were transfected to breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, and ZR-75-1), and also we used normal breast epithelial cell line (hTERT-HME1) to compare RNF2 gene expression level. After 48-72 hours post-transfection, mRNAs were isolated from the cells, and gene expressions were measured by RT-qPCR after from cDNA syntheses. We observed that RNF2 was highly expressed in SK-BR-3 and MDA-MB-231 cell lines opposite to MCF-7 and ZR-75-1 cell lines. RNF2 was downregulated 5, 5 and 7 fold by miR17-5p, miR20a-5p and miR106b-5p respectively in MCF-7. However, in SK-BR-3 and ZR-75-1 cell lines, miRNAs did not affect significantly RNF2 gene expression level. miR20a-5p decreased RNF2 3 fold and miR17-5p and miR106b-5p did not affect MDA-MB-231. After gene expression analysis, we performed metastasis and invasion assay in MCF-7 cells. For metastasis, we used both wound healing assay and Transwell Cell Migration Assay, and we used Transwell Cell Invasion Assay for invasion. The data of this assay showed that miR17-5p and miR20a-5p decreased both invasion and metastasis level, but miR106b-5p has no effect. We would like to conclude that RNF2 can be targeted by miR17-5p, miR20a-5p and miR106b-5p in MCF-7 cells and also RNF2, which is one of the upregulated genes in aggressive tumor, can be decreased by using these miRNAs. In future, we would like to confirm these results at the protein level and also whether these miRNAs are direct target of RNF2 or not.Keywords: breast cancer, epigenetic, microRNAs, RNF2
Procedia PDF Downloads 1804348 Effect of Media Osmolarity on Vi Biosynthesis on Salmonella enterica serovar Typhi Strain C6524 Cultured on Batch System
Authors: Dwi Arisandi Wijaya, Ernawati Arifin Giri-Rachman, Neni Nurainy
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Typhoid fever disease can be prevented by using a polysaccharide-based vaccine Vi which is a virulence factor of S.typhi. To produce high yield Vi polysaccharide from bacteria, it is important to know the biosynthesis of Vi polysaccharide and the regulators involved. In the In vivo condition, S. typhi faces different osmolarity, and the bacterial two-component system OmpR-EnvZ, regulate by up and down Capsular Vi polysaccharide biosynthesis. A high yielded Vi Polysaccharide strain, S. typhi strain C6524 used to study the effect of media osmolarity on Vi polysaccharide biosynthesis and the osmoregulation pattern of S. typhi strain C6524. The methods were performed by grown S. typhi strain C6524 grown on medium with 50 mM, 100 mM, and 150 mM osmolarity with the batch system. Vi polysaccharide concentration was measured by ELISA method. For further investigation of the osmoregulation pattern of strain C6524, the osmoregulator gene, OmpR, has been isolated and sequenced using the specific primer of the OmpR gene. Nucleotide sequence analysis is done with BLAST and Lallign. Amino Acid sequence analysis is done with Prosite and Multiple Sequence Alignment. The results of cultivation showed the average content of polysaccharide Vi for 50 mM, 100 mM, and 150 mM osmolarities 11.49 μg/mL, 12.06 μg/mL, and 14.53 μg/mL respectively. Analysis using Anova stated that the osmolarity treatment of 150 mM significantly affects Vi content. Analysis of nucleotide sequences shows 100% identity between S. typhi strain C6524 and Ty2. Analysis of amino acid sequences shows that the OmpR response regulator protein of the C6524 strain also has a α4-β5-α5 motif which is important for the regulatory activation system when phosphorylation occurs by domain kinase. This indicates that the regulator osmolarity response of S. typhi strain C6524 has no difference with the response regulator owned by S. typhi strain Ty2. A high Vi response rate in the 150 mM osmolarity treatment requires further research for RcsB-RcsC, another two-component system involved in Vi Biosynthesis.Keywords: osmoregulator, OmpR, Salmonella, Vi polysaccharide
Procedia PDF Downloads 1984347 Expression of Human Papillomavirus Type 18 L1 Virus-Like Particles in Methylotropic Yeast, Pichia Pastoris
Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko
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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology
Procedia PDF Downloads 4074346 In Vivo Investigation of microRNA Expression and Function at the Mammalian Synapse by AGO-APP
Authors: Surbhi Surbhi, Andrea Erni, Gunter Meister, Harold Cremer, Christophe Beclin
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MicroRNAs (miRNAs) are short 20-23 nucleotide long non-coding RNAs; there are 2605 miRNA in humans and 1936 miRNA in mouse in total (miRBase). The nervous system expresses the most abundant miRNA and most diverse. MiRNAs play a role in many steps during neurogenesis, like cell proliferation, differentiation, neural patterning, axon pathfinding, etc. Moreover, in vitro studies suggested a role in the regulation of local translation at the synapse, thus controlling neuronal plasticity. However, due to the specific structure of miRNA molecules, an in-vivo confirmation of the general role of miRNAs in the control of neuronal plasticity is still pending. For example, their small size and their high level of sequence homology make difficult the analysis of their cellular and sub-cellular localization in-vivo by in-situ hybridization. Moreover, it was found that only 40% of the expressed miRNA molecules in a cell are included in RNA-Induced Silencing Complexes (RISC) and, therefore, involved in inhibitory interactions while the rest is silent. Definitively, the development of new tools is needed to have a better understanding of the cellular function of miRNAs, in particular their role in neuronal plasticity. Here we describe a new technique called in-vivo AGO-APP designed to investigate miRNA expression and function in-vivo. This technique is based on the expression of a small peptide derived from the human RISC-complex protein TNRC6B, called T6B, which binds all known Argonaute (Ago) proteins with high affinity allowing the efficient immunoprecipitation of AGO-bound miRNAs. We have generated two transgenic mouse lines conditionally expressing T6B either ubiquitously in the cell or targeted at the synapse. A comparison of the repertoire of miRNAs immuno-precipitated from mature neurons of both mouse lines will provide us with a list of miRNAs showing a specific activity at the synapse. The physiological role of these miRNAs will be subsequently addressed through gain and loss of function experiments.Keywords: RNA-induced silencing complexes, TNRC6B, miRNA, argonaute, synapse, neuronal plasticity, neurogenesis
Procedia PDF Downloads 1314345 A Greener Approach for the Recovery of Proteins from Meat Industries
Authors: Jesus Hernandez, Zead Elzoeiry, Md. S. Islam, Abel E. Navarro
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The adsorption of bovine serum albumin (BSA) and human hemoglobin (Hb) on naturally-occurring adsorbents was studied to evaluate the potential recovery of proteins from meat industry residues. Spent peppermint tea (PM), powdered purple corn cob (PC), natural clay (NC) and chemically-modified clay (MC) were investigated to elucidate the effects of pH, adsorbent dose, initial protein concentration, presence of salts and heavy metals. Equilibrium data were fitted according to isotherm models, reporting a maximum adsorption capacity at pH 8 of 318 and 344 mg BSA/g of PM and NC, respectively. Moreover, Hb displayed maximum adsorption capacity at pH 5 of 125 and 143 mg/g of PM and PC, respectively. Hofmeister salt effect was only observed for PM/Hb system. Salts tend to decrease protein adsorption, and the presence of Cu(II) ions had negligible impacts on the adsorption onto NC and PC. Desorption experiments confirmed that more than 85% of both proteins can be recovered with diluted acids and bases. SEM, EDX, and TGA analyses demonstrated that the adsorbents have favorable morphological and mechanical properties. The long-term goal of this study aims to recover soluble proteins from industrial wastewaters to produce animal food or any protein-based product.Keywords: adsorption, albumin, clay, hemoglobin, spent peppermint leaf
Procedia PDF Downloads 1034344 The Co-Existence of Multidominance and Movement in the Syntax of Chinese Bi-Comparatives
Authors: Yaqing Hu
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This paper puts forward a syntactic analysis involving multidominance and rightward movement in Chinese bi-comparatives, as in 'Yuehan bi Mali gao (John is taller than Mary).' It is argued here that the predicate of comparison is a shared constituent in two small clauses, namely one for the target and one for the standard; and then it moves rightward to form a degree phrase with the comparative morpheme. This proposal comes from four aspects. First, the example above can also be expressed in this way, 'A: Yuehan he Mali, shui gao? (John and Mary, who is taller?) B: Yuehan gao./Yuehan geng gao. (John is taller).' This shows that the gradable adjective is predicated of the target. In addition, according to a constraint on Chinese bi-comparatives, namely the target and the standard must be arguments of the predicate simultaneously, it is not unreasonable to assume that the gradable adjective may also be predicated of the standard. Second, subcomparatives are totally disallowed in Chinese, as in '*zhe-zhang zhuozi bi zhe-zhang yizi kuan chang. (This table is longer than this chair is wide.)' In order to save it from ungrammaticality, the target and the standard should be compared along the same dimension denoted by the gradable adjective. It may follow that in Chinese comparatives, having equal roles in the same eventuality, the target and the standard bear the same thematic relationship with the predicate of comparison. Third, verb-copy can appear in Chinese bi-comparatives, as in 'Yuehan qi ma bi Mali qi ma qi de kuai. (John rides horses faster than Mary does.)' The predicate qi seems to form a small clause with both the target and the standard. This might be supporting evidence that both the target and the standard share the predicate of comparison. Fourth, Chinese comparatives do have comparative morphemes, as in 'Yuehan bi Mali geng gao. (John is taller than Mary)', which is semantically equivalent to the first example above. Thus, it follows that one feature of Chinese comparative morphemes is that they can remain overt or covert in the syntax, which will not affect semantics. This further shows that comparative morphemes in bi-comparatives may not be able to saturate the degree argument denoted by the predicate of comparison due to its optionality in the structure. These four aspects present a challenge to the Direct Analysis used in Chinese comparatives since this approach would presume that the target and the standard somehow show independency with the predicate in the syntax. Meanwhile, this study also rejects the previous analysis of multidomiance in bi-comparatives in which the degree phrase comprised of the comparative morpheme and the gradable adjective may be shared by the standard when the comparative morpheme is covert. This syntactic analysis proposed in this study will therefore offer a different perspective of how to treat degree phrase in Chinese comparatives and may offer evidence to argue whether there is degree phrase movement in bi-comparatives as in its English counterparts.Keywords: Chinese comparatives, degree phrase, movement, multidominance, syntactic analysis
Procedia PDF Downloads 3294343 Establishment of Farmed Fish Welfare Biomarkers Using an Omics Approach
Authors: Pedro M. Rodrigues, Claudia Raposo, Denise Schrama, Marco Cerqueira
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Farmed fish welfare is a very recent concept, widely discussed among the scientific community. Consumers’ interest regarding farmed animal welfare standards has significantly increased in the last years posing a huge challenge to producers in order to maintain an equilibrium between good welfare principles and productivity, while simultaneously achieve public acceptance. The major bottleneck of standard aquaculture is to impair considerably fish welfare throughout the production cycle and with this, the quality of fish protein. Welfare assessment in farmed fish is undertaken through the evaluation of fish stress responses. Primary and secondary stress responses include release of cortisol and glucose and lactate to the blood stream, respectively, which are currently the most commonly used indicators of stress exposure. However, the reliability of these indicators is highly dubious, due to a high variability of fish responses to an acute stress and the adaptation of the animal to a repetitive chronic stress. Our objective is to use comparative proteomics to identify and validate a fingerprint of proteins that can present an more reliable alternative to the already established welfare indicators. In this way, the culture conditions will improve and there will be a higher perception of mechanisms and metabolic pathway involved in the produced organism’s welfare. Due to its high economical importance in Portuguese aquaculture Gilthead seabream will be the elected species for this study. Protein extracts from Gilthead Seabream fish muscle, liver and plasma, reared for a 3 month period under optimized culture conditions (control) and induced stress conditions (Handling, high densities, and Hipoxia) are collected and used to identify a putative fish welfare protein markers fingerprint using a proteomics approach. Three tanks per condition and 3 biological replicates per tank are used for each analisys. Briefly, proteins from target tissue/fluid are extracted using standard established protocols. Protein extracts are then separated using 2D-DIGE (Difference gel electrophoresis). Proteins differentially expressed between control and induced stress conditions will be identified by mass spectrometry (LC-Ms/Ms) using NCBInr (taxonomic level - Actinopterygii) databank and Mascot search engine. The statistical analysis is performed using the R software environment, having used a one-tailed Mann-Whitney U-test (p < 0.05) to assess which proteins were differentially expressed in a statistically significant way. Validation of these proteins will be done by comparison of the RT-qPCR (Quantitative reverse transcription polymerase chain reaction) expressed genes pattern with the proteomic profile. Cortisol, glucose, and lactate are also measured in order to confirm or refute the reliability of these indicators. The identified liver proteins under handling and high densities induced stress conditions are responsible and involved in several metabolic pathways like primary metabolism (i.e. glycolysis, gluconeogenesis), ammonia metabolism, cytoskeleton proteins, signalizing proteins, lipid transport. Validition of these proteins as well as identical analysis in muscle and plasma are underway. Proteomics is a promising high-throughput technique that can be successfully applied to identify putative welfare protein biomarkers in farmed fish.Keywords: aquaculture, fish welfare, proteomics, welfare biomarkers
Procedia PDF Downloads 1564342 The Impact of Nutritional Education for Peritoneal Dialysis Patients in Mongolia
Authors: Sanchir Erdenebayar, Namuuntsetseg Oyunbaatar
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Objectives: Peritoneal dialysis treatment is one of the important forms of kidney replacement therapy, and it has recently developed instantly in Mongolia for the past five years. Currently, more than 120 patients undergo peritoneal dialysis nationwide. These patients lack nutritional education, which predisposes them to protein deficiency and further impairs their quality of life. However, there is no study which is conducted among those about their dietary in Mongolia. Therefore, integrated nutrition information and educating them about dietary patterns to follow are urgently needed for PD patients. Methods: A cross-sectional study was carried out on 45 patients aged between 18 and 60 years who were undergoing CAPD at the biggest Medvic dialysis center in Ulaanbaatar. The knowledge of nutrition and food intake is assessed by interview based on a validated questionnaire prepared from KDIGO guidelines, semi-FFQ and a 24-hour dietary recall method. In addition, a biochemical blood test that includes total protein, albumin, calcium, phosphorus, potassium, and hemoglobin is used for an assessment of the patient’s current nutritional status. Results: Knowledge of nutritional status for CAPD was great, with 21.4% of patients and 78.65% having poor nutrition knowledge. The rate of mild to moderate malnutrition was 48.8% among research participants. Serum albumin was 38.4 ± 4.7 g/L, and total protein was 67.3±7.5g/l. Patients met 62.5± 26.5% of their daily intake nutritional requirement for calories and 72±40% of their nutritional requirement for protein. All patients’ energy intake was significantly /1328±304kcal/ lower than the energy requirement (2124±378kcal). Only 14.2% met the recommended dietary protein intake recommended to them of greater than 1.2 g/kg. Conclusions: As was established before, nutritional education has a vital positive impact on the health and nutritional status of peritoneal dialysis patients. The results of this study show that nutritional education programs are not enough adequate in peritoneal dialysis patients. There is a crucial priority to establish nutritional educational programs and guidelines for PD patients in Mongolia.Keywords: renal diet, peritoneal dialysis, nutrition education, CKD diet
Procedia PDF Downloads 624341 Effects of Boron Compounds in Rabbits Fed High Protein and Energy Diet: A Metabolomic and Transcriptomic Approach
Authors: Nuri Başpınar, Abdullah Başoğlu, Özgür Özdemir, Çağlayan Özel, FundaTerzi, Özgür Yaman
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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like nonalcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family were performed. LDL-cholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARα, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to precede in this effectiveness.Keywords: high protein and energy diet, boron, metabolomics, transcriptomic
Procedia PDF Downloads 6274340 Development of Non-frozen Vegan Burger Patty Using Tender Jackfruit (Artocarpus Heterophyllus) as a Meat Substitute: Evaluation of Textural, Physico-Chemical, and Sensory Characteristics
Authors: O. D. A. N. Perera, H. G. Wanigasinghe
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Tender jackfruit is an underutilized biomass, which still has a good consumer demand. Valorization of this ingredient into meat analog would obtain greater consumer acceptance due to concerns about health, the environment, and living sustainably of mankind have increased significantly in this decade, opening the market for meat substitutes. The objective of this research was to create a plant-based meat substitute with a structure similar to meat products. In this study, three different combinations of tender jackfruit were used to create vegan burger patties, which were then examined for their textural, physico-chemical, and sensory qualities. The developed burger patties have been compared with store-bought chicken patties. The developed vegan burger patties P1, P2, and P3 had a comparable flavor preference to the control and demonstrated considerable general acceptability (p >.05). P3 has a high quantity of protein (17.10 ± 0.02%) and fiber (6.40 ± 0.06%). At the same time, the vegan burger patty resulted in less fat, high fiber, and high protein which meets the vegan consumer requirements.Keywords: underutilized, high fibre, soya protein isolate, cooking yield
Procedia PDF Downloads 654339 ELISA Based hTSH Assessment Using Two Sensitive and Specific Anti-hTSH Polyclonal Antibodies
Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati
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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.Keywords: hTSH, bioinformatics, protein expression, cross reactivity
Procedia PDF Downloads 1894338 Deproteination and Demineralization of Shrimp Waste Using Lactic Acid Bacteria for the Production of Crude Chitin and Chitosan
Authors: Farramae Francisco, Rhoda Mae Simora, Sharon Nunal
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Deproteination and demineralization efficiencies of shrimp waste using two Lactobacillus species treated with different carbohydrate sources for chitin production, its chemical conversion to chitosan and the quality of chitin and chitosan produced were determined. Using 5% glucose and 5% cassava starch as carbohydrate sources, pH slightly increased from the initial pH of 6.0 to 6.8 and 7.2, respectively after 24 h and maintained their pH at 6.7 to 7.3 throughout the treatment period. Demineralization (%) in 5 % glucose and 5 % cassava was highest during the first day of treatment which was 82% and 83%, respectively. Deproteination (%) was highest in 5% cassava starch on the 3rd day of treatment at 84.4%. The obtained chitin from 5% cassava and 5% glucose had a residual ash and protein below 1% and solubility of 59% and 44.3%, respectively. Chitosan produced from 5% cassava and 5% glucose had protein content below 0.05%; residual ash was 1.1% and 0.8%, respectively. Chitosan solubility and degree of deacetylation were 56% and 33% in 5% glucose and 48% and 29% in 5% cassava, respectively. The advantage this alternative technology offers over that of chemical extraction is large reduction in chemicals needed thus less effluent production and generation of a protein-rich liquor, although the demineralization process should be improved to achieve greater degree of deacetylation.Keywords: alternative carbon source, bioprocessing, lactic acid bacteria, waste utilization
Procedia PDF Downloads 4854337 Anticancer Activity of Calyx of Diospyros kaki Thunb. through Downregulation of Cyclin D1 Protein Level in Human Colorectal Cancer Cells
Authors: Jin Boo Jeong
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In this study, we elucidated anti-cancer activity and potential molecular mechanism of DKC against human colorectal cancer cells. DKC-E70 suppressed the proliferation of human colorectal cancer cell lines such as HCT116, SW480, LoVo and HT-29. Although DKC-E70 decreased cyclin D1 expression in protein and mRNA level, decreased level of cyclin D1 protein by DKC-E70 occurred at the earlier time than that of cyclin D1 mRNA, which indicates that DKC-E70-mediated downregulation of cyclin D1 protein may be a consequence of the induction of degradation and transcriptional inhibition of cyclin D1. In cyclin D1 degradation, we found that cyclin D1 downregulation by DKC-E70 was attenuated in presence of MG132. In addition, DKC-E70 phosphorylated threonine-286 (T286) of cyclin D1 and T286A abolished cyclin D1 downregulation by DKC-E70. We also observed that DKC-E70-mediated T286 phosphorylation and subsequent cyclin D1 degradation was blocked in presence of the inhibitors of ERK1/2, p38 or GSK3β. In cyclin D1 transcriptional inhibition, DKC-E70 inhibited the expression of β-catenin and TCF4, and β–catenin/TCF-dependent luciferase activity. Our results suggest that DKC-E70 may downregulate cyclin D1 as one of the potential anti-cancer targets through cyclin D1 degradation by T286 phosphorylation dependent on ERK1/2, p38 or GSK3β, and cyclin D1 transcriptional inhibition through Wnt signaling. From these findings, DKC-E70 has potential to be a candidate for the development of chemoprevention or therapeutic agents for human colorectal cancer. This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2016R1D1A3B03931713).Keywords: anticancer, calyx of persimmon, cyclin D1, Diospyros kaki Thunb., human colorectal cancer
Procedia PDF Downloads 3124336 Proactive Competence Management for Employees: A Bottom-up Process Model for Developing Target Competence Profiles Based on the Employee's Tasks
Authors: Maximilian Cedzich, Ingo Dietz Von Bayer, Roland Jochem
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In order for industrial companies to continue to succeed in dynamic, globalized markets, they must be able to train their employees in an agile manner and at short notice in line with the exogenous conditions that arise. For this purpose, it is indispensable to operate a proactive competence management system for employees that recognizes qualification needs timely in order to be able to address them promptly through qualification measures. However, there are hardly any approaches to be found in the literature that includes systematic, proactive competence management. In order to help close this gap, this publication presents a process model that systematically develops bottom-up, future-oriented target competence profiles based on the tasks of the employees. Concretely, in the first step, the tasks of the individual employees are examined for assumed future conditions. In other words, qualitative scenarios are considered for the individual tasks to determine how they are likely to change. In a second step, these scenario-based future tasks are translated into individual future-related target competencies of the employee using a matrix of generic task properties. The final step pursues the goal of validating the target competence profiles formed in this way within the framework of a management workshop. This process model provides industrial companies with a tool that they can use to determine the competencies required by their own employees in the future and compare them with the actual prevailing competencies. If gaps are identified between the target and the actual, these qualification requirements can be closed in the short term by means of qualification measures.Keywords: dynamic globalized markets, employee competence management, industrial companies, knowledge management
Procedia PDF Downloads 1894335 Study on Acoustic Source Detection Performance Improvement of Microphone Array Installed on Drones Using Blind Source Separation
Authors: Youngsun Moon, Yeong-Ju Go, Jong-Soo Choi
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Most drones that currently have surveillance/reconnaissance missions are basically equipped with optical equipment, but we also need to use a microphone array to estimate the location of the acoustic source. This can provide additional information in the absence of optical equipment. The purpose of this study is to estimate Direction of Arrival (DOA) based on Time Difference of Arrival (TDOA) estimation of the acoustic source in the drone. The problem is that it is impossible to measure the clear target acoustic source because of the drone noise. To overcome this problem is to separate the drone noise and the target acoustic source using Blind Source Separation(BSS) based on Independent Component Analysis(ICA). ICA can be performed assuming that the drone noise and target acoustic source are independent and each signal has non-gaussianity. For maximized non-gaussianity each signal, we use Negentropy and Kurtosis based on probability theory. As a result, we can improve TDOA estimation and DOA estimation of the target source in the noisy environment. We simulated the performance of the DOA algorithm applying BSS algorithm, and demonstrated the simulation through experiment at the anechoic wind tunnel.Keywords: aeroacoustics, acoustic source detection, time difference of arrival, direction of arrival, blind source separation, independent component analysis, drone
Procedia PDF Downloads 1624334 Phytomolecules Intervening Inflammation in IgA Nephropathy: A Possible Therapeutic Approach
Authors: Rajiv Jash, Himangshusekhar Maji
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Phytomolecules have long been associated with the effective treatment of various disorders since ages. This study focuses on identifying the immunomodulatory pure molecules isolated from plants, which can be studied for their effect in alleviating IgAN. All the phytomolecules mentioned here have inflammation-reducing properties, and IgAN, being an autoimmune disease, can be a good target of these phytomolecules. Various pathological pathways of IgA nephropathy can be targeted with these phytomolecules, and this study is an effort to find out the rationale behind the choice of the molecules based on their ability to target the effector molecules of those pathological pathways.Keywords: IgAN, fibrosis, inflammation, ESRD, TGFβ
Procedia PDF Downloads 974333 A Study on Information Structure in the Vajrachedika-Prajna-paramita Sutra and Translation Aspect
Authors: Yoon-Cheol Park
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This research focuses on examining the information structures in the old Chinese character-Korean translation of the Vajrachedika-prajna-paramita sutra. The background of this research comes from the fact that there were no previous researches which looked into the information structures in the target text of the Vajrachedika-prajna-paramita sutra by now. The existing researches on the Buddhist scripture translation mainly put weight on message conveyance by literal and semantic translation methods. But the message conveyance from one language to another has a necessity to be delivered with equivalent information structure. Thus, this research is intended to investigate on the flow of old and new information in the target text of Buddhist scripture, compared with source text. The Vajrachedika-prajna-paramita sutra unlike other Buddhist scriptures is composed of conversational structures between Buddha and his disciple, Suboli. This implies that the information flow can be changed by utterance context and some propositions. So, this research tries to analyze the flow of old and new information within the source and target text. As a result of analysis, this research can discover the following facts; firstly, there are the differences of the information flow in the message conveyance between the old Chinese character and Korean by language features. The old Chinese character reveals that old-new information flow is developed, while Korean indicates new-old information flow because of word order. Secondly, the source text of the Vajrachedika-prajna-paramita sutra includes abstruse terminologies, jargon and abstract words. These make influence on the target text and cause the change of the information flow. But the repetitive expressions of these words provide the old information in the target text. Lastly, the Vajrachedika-prajna-paramita sutra offers the expository structure from conversations between Buddha and Suboli. It means that the information flow is developed in the way of explaining specific subjects and of paraphrasing unfamiliar phrases and expressions. From the results of analysis above, this research can verify that the information structures in the target text of the Vajrachedika-prajna-paramita sutra are changed by specific subjects and terminologies, developed with the new-old information flow by repetitive expressions or word order and reveal the information structures familiar to target culture. It also implies that the translation of the Vajrachedika-prajna-paramita sutra as a religious book needs the message conveyance to take into account the information structures of two languages.Keywords: abstruse terminologies, the information structure, new and old information, old Chinese character-Korean translation
Procedia PDF Downloads 3684332 Sensing Mechanism of Nano-Toxic Ions Using Quartz Crystal Microbalance
Authors: Chanho Park, Juneseok You, Kuewhan Jang, Sungsoo Na
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Detection technique of nanotoxic materials is strongly imperative, because nano-toxic materials can harmfully influence human health and environment as their engineering applications are growing rapidly in recent years. In present work, we report the DNA immobilized quartz crystal microbalance (QCM) based sensor for detection of nano-toxic materials such as silver ions, Hg2+ etc. by using functionalization of quartz crystal with a target-specific DNA. Since the mass of a target material is comparable to that of an atom, the mass change caused by target binding to DNA on the quartz crystal is so small that it is practically difficult to detect the ions at low concentrations. In our study, we have demonstrated fast and in situ detection of nanotoxic materials using quartz crystal microbalance. We report the label-free and highly sensitive detection of silver ion for present case, which is a typical nano-toxic material by using QCM and silver-specific DNA. The detection is based on the measurement of frequency shift of Quartz crystal from constitution of the cytosine-Ag+-cytosine binding. It is shown that the silver-specific DNA measured frequency shift by QCM enables the capturing of silver ions below 100pM. The results suggest that DNA-based detection opens a new avenue for the development of a practical water-testing sensor.Keywords: nano-toxic ions, quartz crystal microbalance, frequency shift, target-specific DNA
Procedia PDF Downloads 3204331 Modulation of Fish Allergenicity towards the Production of a Low Allergen Farmed Fish
Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues
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Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.Keywords: fish allergies, fish nutrition, proteomics, aquaculture
Procedia PDF Downloads 1564330 Sensor Registration in Multi-Static Sonar Fusion Detection
Authors: Longxiang Guo, Haoyan Hao, Xueli Sheng, Hanjun Yu, Jingwei Yin
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In order to prevent target splitting and ensure the accuracy of fusion, system error registration is an important step in multi-static sonar fusion detection system. To eliminate the inherent system errors including distance error and angle error of each sonar in detection, this paper uses offline estimation method for error registration. Suppose several sonars from different platforms work together to detect a target. The target position detected by each sonar is based on each sonar’s own reference coordinate system. Based on the two-dimensional stereo projection method, this paper uses real-time quality control (RTQC) method and least squares (LS) method to estimate sensor biases. The RTQC method takes the average value of each sonar’s data as the observation value and the LS method makes the least square processing of each sonar’s data to get the observation value. In the underwater acoustic environment, matlab simulation is carried out and the simulation results show that both algorithms can estimate the distance and angle error of sonar system. The performance of the two algorithms is also compared through the root mean square error and the influence of measurement noise on registration accuracy is explored by simulation. The system error convergence of RTQC method is rapid, but the distribution of targets has a serious impact on its performance. LS method can not be affected by target distribution, but the increase of random noise will slow down the convergence rate. LS method is an improvement of RTQC method, which is widely used in two-dimensional registration. The improved method can be used for underwater multi-target detection registration.Keywords: data fusion, multi-static sonar detection, offline estimation, sensor registration problem
Procedia PDF Downloads 1694329 Production of Mycelial Biomass, Exopolysaccharide, and Enzyme during Solid-State Fermentation of Plant Raw Materials by Medicinal Mushrooms
Authors: Tamar Khardziani, Violeta Berikashvili, Amrosi Chkuaseli, Vladimir Elisashvili
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The main objectives of this proposal are to develop low-cost, innovative, and competitive technologies for the production of mycelial biomass of medicinal mushrooms as a natural food supplement for poultry. To fulfill this task, industrial strains of Lentinus edodes, Ganoderma lucidum, and Pleurotus ostreatus were used in this study. The solid-state fermentation (SSF) of wheat grains, wheat bran, and soy flour was performed in flasks and bags. Among nine mushroom strains, P. ostreatus 2191 appeared to be the most productive in protein biomass accumulation in the SSF of wheat bran. All mushrooms produced exopolysaccharide with the highest yield of 5-8 mg/mL depending on fungal strain and growth substrate. Supplementation of medium with 1% glycerol and 2-4% peptone favored mushroom growth and protein accumulation. Among inorganic nitrogen sources, KNO₃ also provided high biomass and protein production. The SSF of all growth substrates was accompanied by the secretion of cellulase and xylanase activities. The highest CMCase activity (12-13 U/g) was revealed in the cultivation of P. ostreatus 2191 using wheat bran as a growth substrate and ammonium sulfate or yeast extract as a nitrogen source, whereas the highest xylanase activity was detected in the fermentation of soy flour supplemented with peptone. Acknowledgments: This work was supported by the Shota Rustaveli National Science Foundation of Georgia (Grant number STEM-22-2077).Keywords: mushrooms, plant raw materials, fermentation, biomass protein, cellulase
Procedia PDF Downloads 784328 Applicability of Polyisobutylene-Based Polyurethane Structures in Biomedical Disciplines: Some Calcification and Protein Adsorption Studies
Authors: Nihan Nugay, Nur Cicek Kekec, Kalman Toth, Turgut Nugay, Joseph P. Kennedy
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In recent years, polyurethane structures are paving the way for elastomer usage in biology, human medicine, and biomedical application areas. Polyurethanes having a combination of high oxidative and hydrolytic stability and excellent mechanical properties are focused due to enhancing the usage of PUs especially for implantable medical device application such as cardiac-assist. Currently, unique polyurethanes consisting of polyisobutylenes as soft segments and conventional hard segments, named as PIB-based PUs, are developed with precise NCO/OH stoichiometry (∽1.05) for obtaining PIB-based PUs with enhanced properties (i.e., tensile stress increased from ∽11 to ∽26 MPa and elongation from ∽350 to ∽500%). Static and dynamic mechanical properties were optimized by examining stress-strain graphs, self-organization and crystallinity (XRD) traces, rheological (DMA, creep) profiles and thermal (TGA, DSC) responses. Annealing procedure was applied for PIB-based PUs. Annealed PIB-based PU shows ∽26 MPa tensile strength, ∽500% elongation, and ∽77 Microshore hardness with excellent hydrolytic and oxidative stability. The surface characters of them were examined with AFM and contact angle measurements. Annealed PIB-based PU exhibits the higher segregation of individual segments and surface hydrophobicity thus annealing significantly enhances hydrolytic and oxidative stability by shielding carbamate bonds by inert PIB chains. According to improved surface and microstructure characters, greater efforts are focused on analyzing protein adsorption and calcification profiles. In biomedical applications especially for cardiological implantations, protein adsorption inclination on polymeric heart valves is undesirable hence protein adsorption from blood serum is followed by platelet adhesion and subsequent thrombus formation. The protein adsorption character of PIB-based PU examines by applying Bradford assay in fibrinogen and bovine serum albumin solutions. Like protein adsorption, calcium deposition on heart valves is very harmful because vascular calcification has been proposed activation of osteogenic mechanism in the vascular wall, loss of inhibitory factors, enhance bone turnover and irregularities in mineral metabolism. The calcium deposition on films are characterized by incubating samples in simulated body fluid solution and examining SEM images and XPS profiles. PIB-based PUs are significantly more resistant to hydrolytic-oxidative degradation, protein adsorption and calcium deposition than ElastEonTM E2A, a commercially available PDMS-based PU, widely used for biomedical applications.Keywords: biomedical application, calcification, polyisobutylene, polyurethane, protein adsorption
Procedia PDF Downloads 2574327 Effect of Mutagenic Compounds on the Yield of Cultivated Pleurotus Pulmonarius
Authors: Simbiat O. Ayilara-Akande, Soji Fakoya
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Quality and yield are always the target of farmers, including mushroom farmers. This study investigated how better Pleurotus pulmonarius can be obtained with the induction of mutagens into the process of spawn production in order to improve both the quality and the yield. Mushroom spawns were treated with ultraviolet radiation (UV) and hydroxylamine hydrochloride (HA) at different exposure times (2, 6, and 10 minutes) and different concentrations (10, 30, and 50Mm), respectively. The treated spawns were used to cultivate mushrooms on five substrates in the family of Gramineae viz: sorghum, rice, bamboo, sugarcane, and corn straws. Matured fruit bodies were harvested after a few weeks, and their parameters were taken and recorded. This study reveals a significant yield increase in mushroom grown on all the substrates when treated with ultraviolet radiation (UV) for 10 minutes and 6 minutes, respectively. Mushroom spawns treated with hydroxylamine hydrochloride showed a negative correlation in the yield with an increased in mutagen concentration. Hence, Ultraviolet light could be employed to enhance the quality and yield of mushroom production.Keywords: mushroom, protein, mutagens, yield
Procedia PDF Downloads 1484326 Automatic Vowel and Consonant's Target Formant Frequency Detection
Authors: Othmane Bouferroum, Malika Boudraa
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In this study, a dual exponential model for CV formant transition is derived from locus theory of speech perception. Then, an algorithm for automatic vowel and consonant’s target formant frequency detection is developed and tested on real speech. The results show that vowels and consonants are detected through transitions rather than their small stable portions. Also, vowel reduction is clearly observed in our data. These results are confirmed by the observations made in perceptual experiments in the literature.Keywords: acoustic invariance, coarticulation, formant transition, locus equation
Procedia PDF Downloads 2704325 CAP-Glycine Protein Governs Growth, Differentiation, and the Pathogenicity of Global Meningoencephalitis Fungi
Authors: Kyung-Tae Lee, Li Li Wang, Kwang-Woo Jung, Yong-Sun Bahn
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Microtubules are involved in mechanical support, cytoplasmic organization as well as in a number of cellular processes by interacting with diverse microtubule-associated proteins (MAPs), such as plus-end tracking proteins, motor proteins, and tubulin-folding cofactors. A common feature of these proteins is the presence of a cytoskeleton-associated protein-glycine-rich (CAP-Gly) domain, which is evolutionarily conserved and generally considered to bind to α-tubulin to regulate functions of microtubules. However, there has been a dearth of research on CAP-Gly proteins in fungal pathogens, including Cryptococcus neoformans, which causes fatal meningoencephalitis globally. In this study, we identified five CAP-Gly proteins encoding genes in C. neoformans. Among these, Cgp1, encoded by CNAG_06352, has a unique domain structure that has not been reported before in other eukaryotes. Supporting the role of Cpg1 in microtubule-related functions, we demonstrate that deletion or overexpression of CGP1 alters cellular susceptibility to thiabendazole, a microtubule destabilizer, and Cgp1 is co-localized with cytoplasmic microtubules. Related to the cellular functions of microtubules, Cgp1 also governs maintenance of membrane stability and genotoxic stress responses. Furthermore, we demonstrate that Cgp1 uniquely regulates sexual differentiation of C. neoformans with distinct roles in the early and late stage of mating. Our domain analysis reveals that the CAP-Gly domain plays major roles in all the functions of Cgp1. Finally, the cgp1Δ mutant is attenuated in virulence. In conclusion, this novel CAP-Gly protein, Cgp1, has pleotropic roles in regulating growth, stress responses, differentiation and pathogenicity of C. neoformans.Keywords: human fungal pathogen, CAP-Glycine protein, microtubule, meningoencephalitis
Procedia PDF Downloads 3154324 Quantitative Proteome Analysis and Bioactivity Testing of New Zealand Honeybee Venom
Authors: Maryam Ghamsari, Mitchell Nye-Wood, Kelvin Wang, Angela Juhasz, Michelle Colgrave, Don Otter, Jun Lu, Nazimah Hamid, Thao T. Le
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Bee venom, a complex mixture of peptides, proteins, enzymes, and other bioactive compounds, has been widely studied for its therapeutic application. This study investigated the proteins present in New Zealand (NZ) honeybee venom (BV) using bottom-up proteomics. Two sample digestion techniques, in-solution digestion and filter-aided sample preparation (FASP), were employed to obtain the optimal method for protein digestion. Sequential Window Acquisition of All Theoretical Mass Spectra (SWATH–MS) analysis was conducted to quantify the protein compositions of NZ BV and investigate variations in collection years. Our results revealed high protein content (158.12 µg/mL), with the FASP method yielding a larger number of identified proteins (125) than in-solution digestion (95). SWATH–MS indicated melittin and phospholipase A2 as the most abundant proteins. Significant variations in protein compositions across samples from different years (2018, 2019, 2021) were observed, with implications for venom's bioactivity. In vitro testing demonstrated immunomodulatory and antioxidant activities, with a viable range for cell growth established at 1.5-5 µg/mL. The study underscores the value of proteomic tools in characterizing bioactive compounds in bee venom, paving the way for deeper exploration into their therapeutic potentials. Further research is needed to fractionate the venom and elucidate the mechanisms of action for the identified bioactive components.Keywords: honeybee venom, proteomics, bioactivity, fractionation, swath-ms, melittin, phospholipase a2, new zealand, immunomodulatory, antioxidant
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