Search results for: vascular cell adhesion molecules
695 Assessing Brain Targeting Efficiency of Ionisable Lipid Nanoparticles Encapsulating Cas9 mRNA/gGFP Following Different Routes of Administration in Mice
Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal
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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.Keywords: CRISPR, nanoparticles, brain diseases, administration routes
Procedia PDF Downloads 104694 Human Leukocyte Antigen Class 1 Phenotype Distribution and Analysis in Persons from Central Uganda with Active Tuberculosis and Latent Mycobacterium tuberculosis Infection
Authors: Helen K. Buteme, Rebecca Axelsson-Robertson, Moses L. Joloba, Henry W. Boom, Gunilla Kallenius, Markus Maeurer
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Background: The Ugandan population is heavily affected by infectious diseases and Human leukocyte antigen (HLA) diversity plays a crucial role in the host-pathogen interaction and affects the rates of disease acquisition and outcome. The identification of HLA class 1 alleles and determining which alleles are associated with tuberculosis (TB) outcomes would help in screening individuals in TB endemic areas for susceptibility to TB and to predict resistance or progression to TB which would inevitably lead to better clinical management of TB. Aims: To be able to determine the HLA class 1 phenotype distribution in a Ugandan TB cohort and to establish the relationship between these phenotypes and active and latent TB. Methods: Blood samples were drawn from 32 HIV negative individuals with active TB and 45 HIV negative individuals with latent MTB infection. DNA was extracted from the blood samples and the DNA samples HLA typed by the polymerase chain reaction-sequence specific primer method. The allelic frequencies were determined by direct count. Results: HLA-A*02, A*01, A*74, A*30, B*15, B*58, C*07, C*03 and C*04 were the dominant phenotypes in this Ugandan cohort. There were differences in the distribution of HLA types between the individuals with active TB and the individuals with LTBI with only HLA-A*03 allele showing a statistically significant difference (p=0.0136). However, after FDR computation the corresponding q-value is above the expected proportion of false discoveries (q-value 0.2176). Key findings: We identified a number of HLA class I alleles in a population from Central Uganda which will enable us to carry out a functional characterization of CD8+ T-cell mediated immune responses to MTB. Our results also suggest that there may be a positive association between the HLA-A*03 allele and TB implying that individuals with the HLA-A*03 allele are at a higher risk of developing active TB.Keywords: HLA, phenotype, tuberculosis, Uganda
Procedia PDF Downloads 406693 Optimization for Guide RNA and CRISPR/Cas9 System Nanoparticle Mediated Delivery into Plant Cell for Genome Editing
Authors: Andrey V. Khromov, Antonida V. Makhotenko, Ekaterina A. Snigir, Svetlana S. Makarova, Natalia O. Kalinina, Valentin V. Makarov, Mikhail E. Taliansky
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Due to its simplicity, CRISPR/Cas9 has become widely used and capable of inducing mutations in the genes of organisms of various kingdoms. The aim of this work was to develop applications for the efficient modification of DNA coding sequences of phytoene desaturase (PDS), coilin and vacuolar invertase (Solanum tuberosum) genes, and to develop a new nanoparticles carrier efficient technology to deliver the CRISPR/Cas9 system for editing the plant genome. For each of the genes - coilin, PDS and vacuolar invertase, five single RNA guide (sgRNAs) were synthesized. To determine the most suitable nanoplatform, two types of NP platforms were used: magnetic NPs (MNPS) and gold NPs (AuNPs). To test the penetration efficiency, they were functionalized with fluorescent agents - BSA * FITS and GFP, as well as labeled Cy3 small-sized RNA. To measure the efficiency, a fluorescence and confocal microscopy were used. It was shown that the best of these options were AuNP - both in the case of proteins and in the case of RNA. The next step was to check the possibility of delivering components of the CRISPR/Cas9 system to plant cells for editing target genes. AuNPs were functionalized with a ribonucleoprotein complex consisting of Cas9 and corresponding to target genes sgRNAs, and they were biolistically bombarded to axillary buds and apical meristems of potato plants. After the treatment by the best NP carrier, potato meristems were grown to adult plants. DNA isolated from this plants was sent to a preliminary fragment of the analysis to screen out the non-transformed samples, and then to the NGS. The present work was carried out with the financial support from the Russian Science Foundation (grant No. 16-16-04019).Keywords: biobombardment, coilin, CRISPR/Cas9, nanoparticles, NPs, PDS, sgRNA, vacuolar invertase
Procedia PDF Downloads 318692 Low-carbon Footprint Diluents in Solvent Extraction for Lithium-ion Battery Recycling
Authors: Abdoulaye Maihatchi Ahamed, Zubin Arora, Benjamin Swobada, Jean-yves Lansot, Alexandre Chagnes
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Lithium-ion battery (LiB) is the technology of choice in the development of electric vehicles. But there are still many challenges, including the development of positive electrode materials exhibiting high cycle ability, high energy density, and low environmental impact. For this latter, LiBs must be manufactured in a circular approach by developing the appropriate strategies to reuse and recycle them. Presently, the recycling of LiBs is carried out by the pyrometallurgical route, but more and more processes implement or will implement the hydrometallurgical route or a combination of pyrometallurgical and hydrometallurgical operations. After producing the black mass by mineral processing, the hydrometallurgical process consists in leaching the black mass in order to uptake the metals contained in the cathodic material. Then, these metals are extracted selectively by liquid-liquid extraction, solid-liquid extraction, and/or precipitation stages. However, liquid-liquid extraction combined with precipitation/crystallization steps is the most implemented operation in the LiB recycling process to selectively extract copper, aluminum, cobalt, nickel, manganese, and lithium from the leaching solution and precipitate these metals as high-grade sulfate or carbonate salts. Liquid-liquid extraction consists in contacting an organic solvent and an aqueous feed solution containing several metals, including the targeted metal(s) to extract. The organic phase is non-miscible with the aqueous phase. It is composed of an extractant to extract the target metals and a diluent, which is usually aliphatic kerosene produced from the petroleum industry. Sometimes, a phase modifier is added in the formulation of the extraction solvent to avoid the third phase formation. The extraction properties of the diluent do not depend only on the chemical structure of the extractant, but it may also depend on the nature of the diluent. Indeed, the interactions between the diluent can influence more or less the interactions between extractant molecules besides the extractant-diluent interactions. Only a few studies in the literature addressed the influence of the diluent on the extraction properties, while many studies focused on the effect of the extractants. Recently, new low-carbon footprint aliphatic diluents were produced by catalytic dearomatisation and distillation of bio-based oil. This study aims at investigating the influence of the nature of the diluent on the extraction properties of three extractants towards cobalt, nickel, manganese, copper, aluminum, and lithium: Cyanex®272 for nickel-cobalt separation, DEHPA for manganese extraction, and Acorga M5640 for copper extraction. The diluents used in the formulation of the extraction solvents are (i) low-odor aliphatic kerosene produced from the petroleum industry (ELIXORE 180, ELIXORE 230, ELIXORE 205, and ISANE IP 175) and (ii) bio-sourced aliphatic diluents (DEV 2138, DEV 2139, DEV 1763, DEV 2160, DEV 2161 and DEV 2063). After discussing the effect of the diluents on the extraction properties, this conference will address the development of a low carbon footprint process based on the use of the best bio-sourced diluent for the production of high-grade cobalt sulfate, nickel sulfate, manganese sulfate, and lithium carbonate, as well as metal copper.Keywords: diluent, hydrometallurgy, lithium-ion battery, recycling
Procedia PDF Downloads 89691 Peptidoglycan Vaccine-On-Chip against a Lipopolysaccharide-Induced Experimental Sepsis Model
Authors: Katerina Bakela, Ioanna Zerva, Irene Athanassakis
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Lipopolysaccharide (LPS) is commonly used in murine sepsis models, which are largely associated with immunosuppression (incretion of MDSCs cells and Tregs, imbalance of inflammatory/anti-inflammatory cytokines) and collapse of the immune system. After adapting the LPS treatment to the needs of locally bred BALB/c mice, the present study explored the protective role of Micrococcus luteus peptidoglycan (PG) pre-activated vaccine-on chip in endotoxemia. The established protocol consisted of five daily intraperitoneal injections of 0.2mg/g LPS. Such protocol allowed longer survival, necessary in the prospect of the therapeutic treatment application. The so-called vaccine-on-chip consists of a 3-dimensional laser micro-texture Si-scaffold loaded with BALB/c mouse macrophages and activated in vitro with 1μg/ml PG, which exert its action upon subcutaneous implantation. The LPS treatment significantly decreased CD4+, CD8+, CD3z+, and CD19+ cells, while increasing myeloid-derived suppressor cells (MDSCs), CD25+, and Foxp3+ cells. These results were accompanied by increased arginase-1 activity in spleen cell lysates and production of IL-6, TNF-a, and IL-18 while acquiring severe sepsis phenotype as defined by the murine sepsis scoring. The in vivo application of PG pre-activated vaccine-on chip significantly decreased the percent of CD11b+, Gr1+, CD25+, Foxp3+ cells, and arginase-1 activity in the spleen of LPS-treated animals, while decreasing IL-6 and TNF-a in the serum, allowing survival to all animals tested and rescuing the severity of sepsis phenotype. In conclusion, these results reveal a promising mode of action of PG pre-activated vaccine-on chip in LPS endotoxemia, strengthening; thus, the use of treatment is septic patients.Keywords: myeloid-derived suppressor cells, peptidoglycan, sepsis, Si-scaffolds
Procedia PDF Downloads 136690 Radiation Skin Decontamination Formulation
Authors: Navneet Sharma, Himanshu Ojha, Dharam Pal Pathak, Rakesh Kumar Sharma
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Radio-nuclides decontamination is an important task because any extra second of deposition leads to deleterious health effects. We had developed and characterise nanoemulsion of p-tertbutylcalix[4]arens using phase inversion temperature (PIT) method and evaluate its decontamination efficacy (DE). The solubility of the drug was determined in various oils and surfactants. Nanoemulsion developed with an HLB value of 11 and different ratios of the surfactants 10% (7:3, w/w), oil (20%, w/w), and double distilled water (70%) were selected. Formulation was characterised by multi-photon spectroscopy and parameters like viscosity, droplet size distribution, zeta potential and stability were optimised. In vitro and Ex vivo decontamination efficacy (DE) was evaluated against Technetium-99m, Iodine-131, and Thallium-201 as radio-contaminants applied over skin of Sprague-Dawley rat and human tissue equivalent model. Contaminants were removed using formulation soaked in cotton swabs at different time intervals and whole body imaging and static counts were recorded using SPECT gamma camera before and after decontamination attempt. Data were analysed using one-way analysis of variance (ANOVA) and was found to be significant (p <0.05). DE of the nanoemulsion loaded with p-tertbutylcalix[4]arens was compared with placebo and recorded to be 88±5%, 90±3% and 89±3% for 99mTc, 131I and 201Tl respectively. Ex-vivo complexation study of p-tertbutylcalix[4]arene nanoemulsion with surrogate nuclides of radioactive thallium and Iodine, were performed on rat skin mounted on Franz diffusion cell using high-resolution sector field inductively coupled plasma mass spectroscopy (HR-SF-ICPMS). More than 90% complexation of the formulation with these nuclides was observed. Results demonstrate that the prepared nanoemulsion formulation was found efficacious for the decontamination of radionuclides from a large contaminated population.Keywords: p-tertbutylcalix[4]arens, skin decontamination, radiological emergencies, nanoemulsion, iodine-131, thallium-201
Procedia PDF Downloads 402689 Large-Area Film Fabrication for Perovskite Solar Cell via Scalable Thermal-Assisted and Meniscus-Guided Bar Coating
Authors: Gizachew Belay Adugna
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Scalable and cost-effective device fabrication techniques are urgent to commercialize the perovskite solar cells (PSCs) for the next photovoltaic (PV) technology. Herein, large-area films of perovskite and hole-transporting materials (HTMs) were developed via a rapid and scalable thermal-assisting bar-coating process in the open air. High-quality and large crystalline grains of MAPbI₃ with homogenous morphology and thickness were obtained on a large-area (10 cm×10 cm) solution-sheared mp-TiO₂/c-TiO₂/FTO substrate. Encouraging photovoltaic performance of 19.02% was achieved for devices fabricated from the bar-coated perovskite film compared to that from the small-scale spin-coated film (17.27%) with 2,2′,7,7′-tetrakis-(N,N-di-p-methoxyphenylamine)-9,9′-spirobifluorene (spiro-OMeTAD) as an HTM whereas a higher power conversion efficiency of 19.89% with improved device stability was achieved by capping a fluorinated (HYC-2) HTM as an alternative to the traditional spiro-OMeTAD. The fluorinated exhibited better molecular packing in the HTM film and deeper HOMO level compared to the nonfluorinated counterpart; thus, improved hole mobility and overall charge extraction in the device were demonstrated. Furthermore, excellent film processability and an impressive PCE of 18.52% were achieved in the large area bar-coated HYC-2 prepared sequentially on the perovskite underlayer in the open atmosphere, compared to the bar-coated spiro-OMeTAD/perovskite (17.51%). This all-solution approach demonstrated the feasibility of high-quality films on a large-area substrate for PSCs, which is a vital step toward industrial-scale PV production.Keywords: perovskite solar cells, hole transporting materials, up-scaling process, power conversion efficiency
Procedia PDF Downloads 72688 Phytosynthesized Iron Nanoparticles Elicited Growth and Biosynthesis of Steviol Glycosides in Invitro Stevia rebaudiana Plant Cultures
Authors: Amir Ali, Laura Yael Mendoza
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The application of nanomaterials is becoming the most effective strategy of elicitation to produce a desirable level of plant biomass with complex medicinal compounds. This study was designed to check the influence of phytosynthesized iron nanoparticles (FeNPs) on physical growth characteristics, antioxidant status, and production of steviol glycosides of in vitro grown Stevia rebaudiana. Effect of different concentrations of iron nanoparticles replacement of iron sulfate in MS medium (stock solution) on invitro stevia plant growth following positive control (MS basal medium), negative control (iron sulfate devoid medium), iron sulfate devoid MS medium and supplemented with FeNPs at different concentrations (5.6 mg/L, 11.2 mg/L, 16.8 mg/L, 22.4 mg/L) was evaluated. The iron deficiency leads to a drastic reduction in plant growth. In contrast, applying FeNPs leads to improvement in plant height, leave diameter, improved leave morphology, etc., in a concentration-dependent manner. Furthermore, the stress caused by FeNPs at 16.8 mg/L in cultures produced higher levels of total phenolic content (3.7 ± 0.042 mg/g dry weight: DW) and total flavonoid content (1.9 ± 0.022 mg/g DW and antioxidant activity (78 ± 4.6%). In addition, plants grown in the presence of FeNPs at 22.4 mg/L resulted in higher enzymatic antioxidant activities (SOD = 3.5 ± 0.042 U/mg; POD = 2.6 ± 0.026 U/mg; CAT = 2.8 ± 0.034 U/mg and APx = 3.6 ± 0.043 U/ mg), respectively. Furthermore, exposure to a higher dose of FeNPs (22.4 mg/L) exhibited the maximum amount of stevioside (stevioside: 4.6 ± 0.058 mg/g (DW) and rebaudioside A: 4.9 ± 0.068 mg/g DW) as compared to other doses. The current investigation confirms the effectiveness of FeNPs in growth media. It offers a suitable prospect for commercially desirable production of S. rebaudiana biomass with higher sweet glycosides profiles in vitro.Keywords: cell culture, stevia, iron nanoparticles, antioxidants
Procedia PDF Downloads 98687 Methylation Analysis of PHF20L1 and DACT2 Gene Promoters in Women with Breast Cancer
Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho
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Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status
Procedia PDF Downloads 301686 Metabolic Profiling of Populus trichocarpa Family 1 UDP-Glycosyltransferases
Authors: Patricia M. B. Saint-Vincent, Anna Furches, Stephanie Galanie, Erica Teixeira Prates, Piet Jones, Nancy Engle, David Kainer, Wellington Muchero, Daniel Jacobson, Timothy J. Tschaplinski
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Uridine diphosphate-glycosyltransferases (UGTs) are enzymes that catalyze sugar transfer to a variety of plant metabolites. UGT substrates, which include plant secondary metabolites involved in lignification, demonstrate new activities and incorporation when glycosylated. Knowledge of UGT function, substrate specificity, and enzyme products is important for plant engineering efforts, especially related to increasing plant biomass through lignification. UGTs in Populus trichocarpa, a biofuel feedstock, and model woody plant, were selected from a pool of gene candidates using rapid prioritization strategies. A functional genomics workflow, consisting of a metabolite genome-wide association study (mGWAS), expression of synthetic codon-optimized genes, and high-throughput biochemical assays with mass spectrometry-based analysis, was developed for determining the substrates and products of previously-uncharacterized enzymes. A total of 40 UGTs from P. trichocarpa were profiled, and the biochemical assay results were compared to predicted mGWAS connections. Assay results confirmed seven of 11 leaf mGWAS associations and demonstrated varying levels of substrate specificity among candidate UGTs. P. trichocarpa UGT substrate processing confirms the role of these newly-characterized enzymes in lignan, flavonoid, and phytohormone metabolism, with potential implications for cell wall biosynthesis, nitrogen uptake, and biotic and abiotic stress responses.Keywords: Populus, metabolite-gene associations, GWAS, bio feedstocks, glycosyltransferase
Procedia PDF Downloads 117685 Hydrometallurgical Processing of a Nigerian Chalcopyrite Ore
Authors: Alafara A. Baba, Kuranga I. Ayinla, Folahan A. Adekola, Rafiu B. Bale
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Due to increasing demands and diverse applications of copper oxide as pigment in ceramics, cuprammonium hydroxide solution for rayon, p-type semi-conductor, dry cell batteries production and as safety disposal of hazardous materials, a study on the hydrometallurgical operations involving leaching, solvent extraction and precipitation for the recovery of copper for producing high grade copper oxide from a Nigerian chalcopyrite ore in chloride media has been examined. At a particular set of experimental parameter with respect to acid concentration, reaction temperature and particle size, the leaching investigation showed that the ore dissolution increases with increasing acid concentration, temperature and decreasing particle diameter at a moderate stirring. The kinetics data has been analyzed and was found to follow diffusion control mechanism. At optimal conditions, the extent of ore dissolution reached 94.3%. The recovery of the total copper from the hydrochloric acid-leached chalcopyrite ore was undertaken by solvent extraction and precipitation techniques, prior to the beneficiation of the purified solution as copper oxide. The purification of the leach liquor was firstly done by precipitation of total iron and manganese using Ca(OH)2 and H2O2 as oxidizer at pH 3.5 and 4.25, respectively. An extraction efficiency of 97.3% total copper was obtained by 0.2 mol/L Dithizone in kerosene at 25±2ºC within 40 minutes, from which ≈98% Cu from loaded organic phase was successfully stripped by 0.1 mol/L HCl solution. The beneficiation of the recovered pure copper solution was carried out by crystallization through alkali addition followed by calcination at 600ºC to obtain high grade copper oxide (Tenorite, CuO: 05-0661). Finally, a simple hydrometallurgical scheme for the operational extraction procedure amenable for industrial utilization and economic sustainability was provided.Keywords: chalcopyrite ore, Nigeria, copper, copper oxide, solvent extraction
Procedia PDF Downloads 397684 Rheological Properties of Thermoresponsive Poly(N-Vinylcaprolactam)-g-Collagen Hydrogel
Authors: Serap Durkut, A. Eser Elcin, Y. Murat Elcin
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Stimuli-sensitive polymeric hydrogels have received extensive attention in the biomedical field due to their sensitivity to physical and chemical stimuli (temperature, pH, ionic strength, light, etc.). This study describes the rheological properties of a novel thermoresponsive poly(N-vinylcaprolactam)-g-collagen hydrogel. In the study, we first synthesized a facile and novel synthetic carboxyl group-terminated thermo-responsive poly(N-vinylcaprolactam)-COOH (PNVCL-COOH) via free radical polymerization. Further, this compound was effectively grafted with native collagen, by utilizing the covalent bond between the carboxylic acid groups at the end of the chains and amine groups of the collagen using cross-linking agent (EDC/NHS), forming PNVCL-g-Col. Newly-formed hybrid hydrogel displayed novel properties, such as increased mechanical strength and thermoresponsive characteristics. PNVCL-g-Col showed low critical solution temperature (LCST) at 38ºC, which is very close to the body temperature. Rheological studies determine structural–mechanical properties of the materials and serve as a valuable tool for characterizing. The rheological properties of hydrogels are described in terms of two dynamic mechanical properties: the elastic modulus G′ (also known as dynamic rigidity) representing the reversible stored energy of the system, and the viscous modulus G″, representing the irreversible energy loss. In order to characterize the PNVCL-g-Col, the rheological properties were measured in terms of the function of temperature and time during phase transition. Below the LCST, favorable interactions allowed the dissolution of the polymer in water via hydrogen bonding. At temperatures above the LCST, PNVCL molecules within PNVCL-g-Col aggregated due to dehydration, causing the hydrogel structure to become dense. When the temperature reached ~36ºC, both the G′ and G″ values crossed over. This indicates that PNVCL-g-Col underwent a sol-gel transition, forming an elastic network. Following temperature plateau at 38ºC, near human body temperature the sample displayed stable elastic network characteristics. The G′ and G″ values of the PNVCL-g-Col solutions sharply increased at 6-9 minute interval, due to rapid transformation into gel-like state and formation of elastic networks. Copolymerization with collagen leads to an increase in G′, as collagen structure contains a flexible polymer chain, which bestows its elastic properties. Elasticity of the proposed structure correlates with the number of intermolecular cross-links in the hydrogel network, increasing viscosity. However, at 8 minutes, G′ and G″ values sharply decreased for pure collagen solutions due to the decomposition of the elastic and viscose network. Complex viscosity is related to the mechanical performance and resistance opposing deformation of the hydrogel. Complex viscosity of PNVCL-g-Col hydrogel was drastically changed with temperature and the mechanical performance of PNVCL-g-Col hydrogel network increased, exhibiting lesser deformation. Rheological assessment of the novel thermo-responsive PNVCL-g-Col hydrogel, exhibited that the network has stronger mechanical properties due to both permanent stable covalent bonds and physical interactions, such as hydrogen- and hydrophobic bonds depending on temperature.Keywords: poly(N-vinylcaprolactam)-g-collagen, thermoresponsive polymer, rheology, elastic modulus, stimuli-sensitive
Procedia PDF Downloads 244683 Dual Electrochemical Immunosensor for IL-13Rα2 and E-Cadherin Determination in Cell, Serum and Tissues from Cancer Patients
Authors: Amira ben Hassine, A. Valverde, V. Serafín, C. Muñoz-San Martín, M. Garranzo-Asensio, M. Gamella, R. Barderas, M. Pedrero, N. Raouafi, S. Campuzano, P. Yáñez-Sedeño, J. M. Pingarrón
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This work describes the development of a dual electrochemical immunosensing platform for accurate determination of two target proteins, IL-13 Receptor α2 (IL-13Rα2) and E-cadherin (E-cad). The proposed methodology is based on the use of sandwich immunosensing approaches (involving horseradish peroxidase-labeled detector antibodies) implemented onto magnetic microbeads (MBs) and amperometric transduction at screen-printed dual carbon electrodes (SPdCEs). The magnetic bioconjugates were captured onto SPdCEs and the amperometric transduction was performed using the H2O2/hydroquinone (HQ) system. Under optimal experimental conditions, the developed bio platform demonstrates linear concentration ranges of 1.0–25 and 5.0-100 ng mL-1, detection limits of 0.28 and 1.04 ng mL-1 for E-cad and IL-13Rα2, respectively, and excellent selectivity against other non-target proteins. The developed immuno-platform also offers a good reproducibility among amperometric responses provided by nine different sensors constructed in the same manner (Relative Standard Deviation values of 3.1% for E-cad and 4.3% for IL-13Rα2). Moreover, obtained results confirm the practical applicability of this bio-platform for the accurate determination of the endogenous levels of both extracellular receptors in colon cancer cells (both intact and lysed) with different metastatic potential and serum and tissues from patients diagnosed with colorectal cancer at different grades. Interesting features in terms of, simplicity, speed, portability and sample amount required to provide quantitative results, make this immuno-platform more compatible than conventional methodologies with the clinical diagnosis and prognosis at the point of care.Keywords: electrochemistry, mmunosensors, biosensors, E-cadherin, IL-13 receptor α2, cancer colorectal
Procedia PDF Downloads 138682 Effect of Microstructure on Wear Resistance of Polycrystalline Diamond Composite Cutter of Bit
Authors: Fanyuan Shao, Wei Liu, Deli Gao
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Polycrystalline diamond composite (PDC) cutter is made of diamond powder as raw material, cobalt metal or non-metallic elements as a binder, mixed with WC cemented carbide matrix assembly, through high temperature and high-pressure sintering. PDC bits with PDC cutters are widely used in oil and gas drilling because of their high hardness, good wear resistance and excellent impact toughness. And PDC cutter is the main cutting tool of bit, which seriously affects the service of the PDC bit. The wear resistance of the PDC cutter is measured by cutting granite with a vertical turret lathe (VTL). This experiment can achieve long-distance cutting to obtain the relationship between the wear resistance of the PDC cutter and cutting distance, which is more closely to the real drilling situation. Load cell and 3D optical profiler were used to obtain the value of cutting forces and wear area, respectively, which can also characterize the damage and wear of the PDC cutter. PDC cutters were cut via electrical discharge machining (EDM) and then flattened and polished. A scanning electron microscope (SEM) was used to observe the distribution of binder cobalt and the size of diamond particles in a diamond PDC cutter. The cutting experimental results show that the wear area of the PDC cutter has a good linear relationship with the cutting distance. Simultaneously, the larger the wear area is and the greater the cutting forces are required to maintain the same cutting state. The size and distribution of diamond particles in the polycrystalline diamond layer have a great influence on the wear resistance of the diamond layer. And PDC cutter with fine diamond grains shows more wear resistance than that with coarse grains. The deep leaching process is helpful to reduce the effect of binder cobalt on the wear resistance of the polycrystalline diamond layer. The experimental study can provide an important basis for the application of PDC cutters in oil and gas drilling.Keywords: polycrystalline diamond compact, scanning electron microscope, wear resistance, cutting distance
Procedia PDF Downloads 200681 Sensing of Cancer DNA Using Resonance Frequency
Authors: Sungsoo Na, Chanho Park
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Lung cancer is one of the most common severe diseases driving to the death of a human. Lung cancer can be divided into two cases of small-cell lung cancer (SCLC) and non-SCLC (NSCLC), and about 80% of lung cancers belong to the case of NSCLC. From several studies, the correlation between epidermal growth factor receptor (EGFR) and NSCLCs has been investigated. Therefore, EGFR inhibitor drugs such as gefitinib and erlotinib have been used as lung cancer treatments. However, the treatments result showed low response (10~20%) in clinical trials due to EGFR mutations that cause the drug resistance. Patients with resistance to EGFR inhibitor drugs usually are positive to KRAS mutation. Therefore, assessment of EGFR and KRAS mutation is essential for target therapies of NSCLC patient. In order to overcome the limitation of conventional therapies, overall EGFR and KRAS mutations have to be monitored. In this work, the only detection of EGFR will be presented. A variety of techniques has been presented for the detection of EGFR mutations. The standard detection method of EGFR mutation in ctDNA relies on real-time polymerase chain reaction (PCR). Real-time PCR method provides high sensitive detection performance. However, as the amplification step increases cost effect and complexity increase as well. Other types of technology such as BEAMing, next generation sequencing (NGS), an electrochemical sensor and silicon nanowire field-effect transistor have been presented. However, those technologies have limitations of low sensitivity, high cost and complexity of data analyzation. In this report, we propose a label-free and high-sensitive detection method of lung cancer using quartz crystal microbalance based platform. The proposed platform is able to sense lung cancer mutant DNA with a limit of detection of 1nM.Keywords: cancer DNA, resonance frequency, quartz crystal microbalance, lung cancer
Procedia PDF Downloads 234680 Study on Shifting Properties of CVT Rubber V-belt
Authors: Natsuki Tsuda, Kiyotaka Obunai, Kazuya Okubo, Hideyuki Tashiro, Yoshinori Yamaji, Hideyuki Kato
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The objective of this study is to investigate the effect of belt stiffness on the performance of the CVT unit, such as the required pulley thrust force and the ratio coverage. The CVT unit consists of the V-grooved pulleys and the rubber CVT belt. The width of the driving pulley groove was controlled by the stepper motor, while that of the driven pulley was controlled by the hydraulic pressure. The generated mechanical power on the motor was transmitted from the driving axis to the driven axis through the CVT unit. The rotational speed and the transmitting torque of both axes were measured by the tachometers and the torque meters attached with these axes, respectively. The transmitted, mechanical power was absorbed by the magnetic powder brake. The thrust force acting on both pulleys and the force between both shafts were measured by the load cell. The back face profile of the rubber CVT belt along with width direction was measured by the 2-dimensional laser displacement meter. This paper found that when the stiffness of the rubber CVT belt in the belt width direction was reduced, the thrust force required for shifting was reduced. Moreover, when the stiffness of the rubber CVT belt in the belt width direction was reduced, the ratio coverage of the CVT unit was reduced. Due to the decrement of stiffness in belt width direction, the excessive concave deformation of belt in pulley groove was confirmed. Because of this excessive concave deformation, apparent wrapping radius of belt would have been reduced. Proposed model could be effectively estimated the difference of ratio coverage due to concave deformation. The proposed model could also be utilized for designing the rubber CVT belt with optimal bending stiffness in width direction.Keywords: CVT, countinuously variable transmission, rubber, belt stiffness, transmission
Procedia PDF Downloads 145679 Efficacy of the Use of Different Teaching Approaches of Math Teachers
Authors: Nilda San Miguel, Elymar Pascual
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The main focus of this study is exploring the effective approaches in teaching Mathematics that is being applied in public schools, s.y. 2018-2019. This research was written as connected output to the district-wide School Learning Action Cell (DISLAC) on Math teaching approaches which was recently conducted in Victoria, Laguna. Fifty-four math teachers coming from 17 schools in Victoria became the respondents of this study. Qualitative method of doing research was applied. Teachers’ responses to the following concerns were gathered, analyzed and interpreted: (1) evaluation of the recently conducted DISLAC, (2) status of the use of different approaches, (3) perception on the effective use of approaches, (4) preference of approach to explore in classroom sessions, (5) factors affecting the choice of approach, (6) difficulties encountered, (7) and perceived benefit to learners. Results showed that the conduct of DISLAC was very highly satisfactory (mean 4.41). Teachers looked at collaborative approach as very highly effective (mean 4.74). Fifty-two percent of the teachers is using collaborative approach, 17% constructivist, 11% integrative, 11% inquiry-based, and 9% reflective. Reflective approach was chosen to be explored by most of the respondents (29%) in future sessions. The difficulties encountered by teachers in using the different approaches are: (1) learners’ difficulty in following instructions, (2) lack of focus, (3) lack of willingness and cooperation, (4) teachers’ lack of mastery in using different approaches, and (5) lack of time of doing visual aids because of time mismanagement. Teachers deemed the use of various teaching approaches can help the learners to have (1) mastery of competency, (2) increased communication, (3) improved confidence, (4) facility in comprehension, and (5) better academic output. The result obtained from this study can be used as an input for SLACs. Recommendations at the end of the study were given to school/district heads and future researchers.Keywords: approaches, collaborative, constructivism, inquiry-based, integrative, reflective
Procedia PDF Downloads 283678 Induction of Cellular and Humoral Immune Responses in BALB/c Mice Immunized With rB2L and rF1L Proteins of Orf Virus Adjuvanted With Alumina Nanoparticles
Authors: Alhaji Modu Bukar, Faez Firdaus Abdullah Jesse, Che Azurahanim Che Abdullah, Mustapha M. Noordin, Mohd-Lila Mohd Azmia
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Orf virus (ORFV) is the causative agent of a proliferative skin lesion known as contagious ecthyma in sheep and goats. Currently used live attenuated vaccines against ORFV infection have been reported to cause severe outbreaks in vaccinated animals. In this study, we investigated the immunogenicity of the B2L and F1L proteins of the virus, which are thought to elicit a protective immune response The 6-week-old 50 female mice were divided into 8 groups: seven experimental groups and one control group. Each animal in the experimental group received an initial immunisation with the nanoparticles or proteins coated with the nanoparticles, followed by two booster immunizations with the same products 14 days apart. Ten days after the last booster inoculation, the mice were either humanely killed or lethally challenged with UPM /HSN-2-ORFV at a dose of 106 TCID50/mL in a volume of 50 μl. The spleen was examined for histopathological changes and quantification of T cells by flow cytometry. On the other hand, the degree of protection of mice from the lethal virus was evaluated by lesion size, weight loss, and histopathological examination of skin and liver. The results showed that mice immunised with rB2L alone, rB2L-Al₂O₃-NPs, rB2L/rF1L, and rB2L/rF1L-Al₂O₃-NPs elicited statistically higher levels of anti-rB2L and/or rF1L-specific IgA/IgG and CD4/CD8 cell immune responses than mice in the control groups (p < 0.01). The vaccine candidate did not exhibit severe skin damage after monitoring histopathology, morbidity, and mortality. Overall, the results suggest that recombinant rB2L and rF1L antigens may be useful universal vaccine candidates against ORFV infections.Keywords: orf virus, antigen nanoparticles, virus, nanoparticles
Procedia PDF Downloads 73677 Prognostic Impact of Pre-transplant Ferritinemia: A Survival Analysis Among Allograft Patients
Authors: Mekni Sabrine, Nouira Mariem
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Background and aim: Allogeneic hematopoietic stem cell transplantation is a curative treatment for several hematological diseases; however, it has a non-negligible morbidity and mortality depending on several prognostic factors, including pre-transplant hyperferritinemia. The aim of our study was to estimate the impact of hyperferritinemia on survivals and on the occurrence of post-transplant complications. Methods: It was a longitudinal study conducted over 8 years and including all patients who had a first allograft. The impact of pretransplant hyperferritinemia (ferritinemia ≥1500) on survivals was studied using the Kaplan Meier method and the COX model for uni- and multivariate analysis. The Khi-deux test and binary logistic regression were used to study the association between pretransplant ferritinemia and post-transplant complications. Results: One hundred forty patients were included with an average age of 26.6 years and a sex ratio (M/F)=1.4. Hyperferritinemia was found in 33% of patients. It had no significant impact on either overall survival (p=0.9) or event -free survival (p=0.6). In multivariate analysis, only the type of disease was independently associated with overall survival (p=0.04) and event-free survival (p=0.002). For post-allograft complications: The occurrence of early documented infections was independently associated with pretransplant hyperferritinemia (p=0.02) and the presence of acute graft versus host disease( GVHD) (p<10-3). The occurrence of acute GVHD was associated with early documented infection (p=0.002) and Cytomegalovirus reactivation (p<10-3). The occurrence of chronic GVHD was associated with the presence of Cytomegalovirus reactivation (p=0.006) and graft source (p=0.009). Conclusion: Our study showed the significant impact of pre-transplant hyperferritinemia on the occurrence of early infections but not on survivals. Early and more accurate assessment iron overload by other tests such as liver magnetic resonance imaging with initiation of chelating treatment could prevent the occurrence of such complications after transplantation.Keywords: allogeneic, transplants, ferritin, survival
Procedia PDF Downloads 67676 QSAR Modeling of Germination Activity of a Series of 5-(4-Substituent-Phenoxy)-3-Methylfuran-2(5H)-One Derivatives with Potential of Strigolactone Mimics toward Striga hermonthica
Authors: Strahinja Kovačević, Sanja Podunavac-Kuzmanović, Lidija Jevrić, Cristina Prandi, Piermichele Kobauri
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The present study is based on molecular modeling of a series of twelve 5-(4-substituent-phenoxy)-3-methylfuran-2(5H)-one derivatives which have potential of strigolactones mimics toward Striga hermonthica. The first step of the analysis included the calculation of molecular descriptors which numerically describe the structures of the analyzed compounds. The descriptors ALOGP (lipophilicity), AClogS (water solubility) and BBB (blood-brain barrier penetration), served as the input variables in multiple linear regression (MLR) modeling of germination activity toward S. hermonthica. Two MLR models were obtained. The first MLR model contains ALOGP and AClogS descriptors, while the second one is based on these two descriptors plus BBB descriptor. Despite the braking Topliss-Costello rule in the second MLR model, it has much better statistical and cross-validation characteristics than the first one. The ALOGP and AClogS descriptors are often very suitable predictors of the biological activity of many compounds. They are very important descriptors of the biological behavior and availability of a compound in any biological system (i.e. the ability to pass through the cell membranes). BBB descriptor defines the ability of a molecule to pass through the blood-brain barrier. Besides the lipophilicity of a compound, this descriptor carries the information of the molecular bulkiness (its value strongly depends on molecular bulkiness). According to the obtained results of MLR modeling, these three descriptors are considered as very good predictors of germination activity of the analyzed compounds toward S. hermonthica seeds. This article is based upon work from COST Action (FA1206), supported by COST (European Cooperation in Science and Technology).Keywords: chemometrics, germination activity, molecular modeling, QSAR analysis, strigolactones
Procedia PDF Downloads 289675 Synthesis of Highly Stable Pseudocapacitors From Secondary Resources
Authors: Samane Maroufi, Rasoul Khayyam Nekouei, Sajjad Mofarah
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Fabrication of the state-of-the-art portable pseudocapacitors with the desired transparency, mechanical flexibility, capacitance, and durability is challenging. In most cases, the fabrication of such devices requires critical elements which are either under the crisis of depletion or their extraction from virgin mineral ores have sever environmental impacts. This urges the use of secondary resources instead of virgin resources in fabrication of advanced devices. In this research, ultrathin films of defect-rich Mn1−x−y(CexLay)O2−δ with controllable thicknesses in the range between 5 nm to 627 nm and transmittance (≈29–100%) have been fabricated via an electrochemical chronoamperometric deposition technique using an aqueous precursor derived during the selective purification of rare earth oxide (REOs) isolated from end-of-life nickel-metal hydride (Ni-MH) batteries. Intercalation/de-intercalation of anionic O2− through the atomic tunnels of the stratified Mn1−x−y(CexLay)O2−δ crystallites was found to be responsible for outstanding areal capacitance of 3.4 mF cm−2 of films with 86% transmittance. The intervalence charge transfer among interstitial Ce/La cations and Mn oxidation states within the Mn1−x−y(CexLay)O2−δ structure resulted in excellent capacitance retention of ≈90% after 16 000 cycles. The synthesised transparent flexible Mn1−x−y(CexLay)O2−δ full-cell pseudocapacitor device possessed the energy and power densities of 0.088 μWh cm⁻² and 843 µW cm⁻², respectively. These values show insignificant changes under vigorous twisting and bending to 45–180° confirming these value-added materials are intriguing alternatives for size-sensitive energy storage devices. This research confirms the feasibility of utilisation of secondary waste resources for the fabrication of high-quality pseudocapacitors with engineered defects with the desired flexibility, transparency, and cycling stability suitable for size-sensitive portable electronic devices.Keywords: pseudocapacitors, energy storage devices, flexible and transparent, sustainability
Procedia PDF Downloads 89674 Development of an Electrochemical Aptasensor for the Detection of Human Osteopontin Protein
Authors: Sofia G. Meirinho, Luis G. Dias, António M. Peres, Lígia R. Rodrigues
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The emerging development of electrochemical aptasen sors has enabled the easy and fast detection of protein biomarkers in standard and real samples. Biomarkers are produced by body organs or tumours and provide a measure of antigens on cell surfaces. When detected in high amounts in blood, they can be suggestive of tumour activity. These biomarkers are more often used to evaluate treatment effects or to assess the potential for metastatic disease in patients with established disease. Osteopontin (OPN) is a protein found in all body fluids and constitutes a possible biomarker because its overexpression has been related with breast cancer evolution and metastasis. Currently, biomarkers are commonly used for the development of diagnostic methods, allowing the detection of the disease in its initial stages. A previously described RNA aptamer was used in the current work to develop a simple and sensitive electrochemical aptasensor with high affinity for human OPN. The RNA aptamer was biotinylated and immobilized on a gold electrode by avidin-biotin interaction. The electrochemical signal generated from the aptamer–target molecule interaction was monitored electrochemically using cyclic voltammetry in the presence of [Fe (CN) 6]−3/− as a redox probe. The signal observed showed a current decrease due to the binding of OPN. The preliminary results showed that this aptasensor enables the detection of OPN in standard solutions, showing good selectivity towards the target in the presence of others interfering proteins such as bovine OPN and bovine serum albumin. The results gathered in the current work suggest that the proposed electrochemical aptasensor is a simple and sensitive detection tool for human OPN and so, may have future applications in cancer disease monitoring.Keywords: osteopontin, aptamer, aptasensor, screen-printed electrode, cyclic voltammetry
Procedia PDF Downloads 433673 Bacterial Interactions of Upper Respiratory Tract Microbiota
Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys
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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.Keywords: nasal, microbiota, S.aureus, microbioal interaction
Procedia PDF Downloads 117672 Development of Fluorescence Resonance Energy Transfer-Based Nanosensor for Measurement of Sialic Acid in vivo
Authors: Ruphi Naz, Altaf Ahmad, Mohammad Anis
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Sialic acid (5-Acetylneuraminic acid, Neu5Ac) is a common sugar found as a terminal residue on glycoconjugates in many animals. Humans brain and the central nervous system contain the highest concentration of sialic acid (as N-acetylneuraminic acid) where these acids play an important role in neural transmission and ganglioside structure in synaptogenesis. Due to its important biological function, sialic acid is attracting increasing attention. To understand metabolic networks, fluxes and regulation, it is essential to be able to determine the cellular and subcellular levels of metabolites. Genetically-encoded fluorescence resonance energy transfer (FRET) sensors represent a promising technology for measuring metabolite levels and corresponding rate changes in live cells. Taking this, we developed a genetically encoded FRET (fluorescence resonance energy transfer) based nanosensor to analyse the sialic acid level in living cells. Sialic acid periplasmic binding protein (sia P) from Haemophilus influenzae was taken and ligated between the FRET pair, the cyan fluorescent protein (eCFP) and Venus. The chimeric sensor protein was expressed in E. coli BL21 (DE3) and purified by affinity chromatography. Conformational changes in the binding protein clearly confirmed the changes in FRET efficiency. So any change in the concentration of sialic acid is associated with the change in FRET ratio. This sensor is very specific to sialic acid and found stable with the different range of pH. This nanosensor successfully reported the intracellular level of sialic acid in bacterial cell. The data suggest that the nanosensors may be a versatile tool for studying the in vivo dynamics of sialic acid level non-invasively in living cellsKeywords: nanosensor, FRET, Haemophilus influenzae, metabolic networks
Procedia PDF Downloads 134671 Optimization of Ultrasound Assisted Extraction of Polysaccharides from Plant Waste Materials: Selected Model Material is Hazelnut Skin
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In this study, optimization of ultrasound assisted extraction (UAE) of hemicellulose based polysaccharides from plant waste material has been studied. Selected material is hazelnut skin. Extraction variables for the operation are extraction time, amplitude and application temperature. Optimum conditions have been evaluated depending on responses such as amount of wet crude polysaccharide, total carbohydrate content and dried sample. Pretreated hazelnut skin powders were used for the experiments. 10 grams of samples were suspended in 100 ml water in a jacketed vessel with additional magnetic stirring. Mixture was sonicated by immersing ultrasonic probe processor. After the extraction procedures, ethanol soluble and insoluble sides were separated for further examinations. The obtained experimental data were analyzed by analysis of variance (ANOVA). Second order polynomial models were developed using multiple regression analysis. The individual and interactive effects of applied variables were evaluated by Box Behnken Design. The models developed from the experimental design were predictive and good fit with the experimental data with high correlation coefficient value (R2 more than 0.95). Extracted polysaccharides from hazelnut skin are assumed to be pectic polysaccharides according to the literature survey of Fourier Transform Spectrometry (FTIR) analysis results. No more change can be observed between spectrums of different sonication times. Application of UAE at optimized condition has an important effect on extraction of hemicellulose from plant material by satisfying partial hydrolysis to break the bounds with other components in plant cell wall material. This effect can be summarized by varied intensity of microjets and microstreaming at varied sonication conditions.Keywords: hazelnut skin, optimization, polysaccharide, ultrasound assisted extraction
Procedia PDF Downloads 333670 Beta-Carotene Attenuates Cognitive and Hepatic Impairment in Thioacetamide-Induced Rat Model of Hepatic Encephalopathy via Mitigation of MAPK/NF-κB Signaling Pathway
Authors: Marawan Abd Elbaset Mohamed, Hanan A. Ogaly, Rehab F. Abdel-Rahman, Ahmed-Farid O.A., Marwa S. Khattab, Reham M. Abd-Elsalam
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Liver fibrosis is a severe worldwide health concern due to various chronic liver disorders. Hepatic encephalopathy (HE) is one of its most common complications affecting liver and brain cognitive function. Beta-Carotene (B-Car) is an organic, strongly colored red-orange pigment abundant in fungi, plants, and fruits. The study attempted to know B-Car neuroprotective potential against thioacetamide (TAA)-induced neurotoxicity and cognitive decline in HE in rats. Hepatic encephalopathy was induced by TAA (100 mg/kg, i.p.) three times per week for two weeks. B-Car was given orally (10 or 20 mg/kg) daily for two weeks after TAA injections. Organ body weight ratio, Serum transaminase activities, liver’s antioxidant parameters, ammonia, and liver histopathology were assessed. Also, the brain’s mitogen-activated protein kinase (MAPK), nuclear factor kappa B (NF-κB), antioxidant parameters, adenosine triphosphate (ATP), adenosine monophosphate (AMP), norepinephrine (NE), dopamine (DA), serotonin (5-HT), 5-hydroxyindoleacetic acid (5-HIAA) cAMP response element-binding protein (CREB) expression and B-cell lymphoma 2 (Bcl-2) expression were measured. The brain’s cognitive functions (Spontaneous locomotor activity, Rotarod performance test, Object recognition test) were assessed. B-Car prevented alteration of the brain’s cognitive function in a dose-dependent manner. The histopathological outcomes supported these biochemical evidences. Based on these results, it could be established that B-Car could be assigned to treat the brain’s neurotoxicity consequences of HE via downregualtion of MAPK/NF-κB signaling pathways.Keywords: beta-carotene, liver injury, MAPK, NF-κB, rat, thioacetamide
Procedia PDF Downloads 155669 Ethical Considerations in In-Utero Gene Editing
Authors: Shruti Govindarajan
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In-utero gene editing with CRISPR-Cas9 opens up new possibilities for treating genetic disorders during pregnancy while still in mother’s womb. By targeting genetic mutations in the early stages of fetal development, this approach could potentially prevent severe conditions—like cystic fibrosis, sickle cell anemia, and muscular dystrophy—from causing harm. CRISPR-Cas9, which allows precise DNA edits, could be delivered into fetal cells through vectors such as adeno-associated viruses (AAVs) or nanoparticles, correcting disease-causing mutations and possibly offering lifelong relief from these disorders. For families facing severe genetic diagnoses, in-utero gene editing could provide a transformative option. However, technical challenges remain, including ensuring that gene editing only targets the intended cells and verifying long-term safety. Ethical considerations are also at the forefront of this technology. The editing of a fetus's genes brings up difficult questions about consent, especially since these genetic changes will affect the child’s entire life without their input. There's also concern over possible unintended side effects, or changes passed down to future generations. Moreover, if used beyond therapeutic purposes, this technology could be misused for ‘enhancements,’ like selecting for certain physical or cognitive traits, raising concerns about inequality and social pressures. In this way, in-utero gene editing brings both exciting potential and complex moral questions. As research progresses, addressing these scientific and ethical concerns will be key to ensuring that this technology is used responsibly, prioritizing safety, fairness, and a focus on alleviating genetic disease. A cautious and inclusive approach, along with clear regulations, will be essential to realizing the benefits of in-utero gene editing while protecting against unintended consequences.Keywords: in-utero gene editing, CRISPR, bioethics, genetic disorder
Procedia PDF Downloads 13668 Nitrate-Induced Biochemical and Histopathological Changes in the Kidney of Rats: Attenuation by Hyparrhenia hirta
Authors: Hanen Bouaziz, Moez Rafrafi, Ghada Ben Salah, Kamel Jamoussi, Tahia Boudawara, Najiba Zeghal
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The present study investigated the protective role of Hyparrhenia hirta against sodium nitrate (NaNO3)-induced nephrotoxicity. A high-performance liquid chromatography coupled with a mass spectrometer (HPLC-MS) method was developed to separate and identify flavonoids in Hyparrhenia hirta. Seven flavonoids were identified as 3-O-methylquercetin, luteolin-7-O-glucoside, luteolin, apigenin-7-O-glucoside, apigenin-8-C-glucoside, luteolin-8-C-glucoside and luteolin-6-C-glucoside. Wistar rats were randomly divided into three groups: a control group and two treated groups during 50 days with NaNO3 administered either alone in drinking water or co-administered with Hyparrhenia hirta. NaNO3 treatment induced a significant increase in plasma levels of creatinine, urea and uric while urinary level decreased significantly. Nephrotoxicity induced by NaNO3 was characterized by significant increase in creatinine clearance. In parallel, a significant increase in malondialdehyde level along with a concomitant decrease in total glutathione content and superoxide dismutase, catalase and glutathione peroxidase activities were observed in the kidney after NaNO3 treatment. The histopathological changes in kidney after NaNO3 administration were shrunken. There were renal tubule cell degeneration and infiltration of mononuclear cells. Most glomeruli revealed shrinkage, a wide capsular space and a peri-glomerular mononuclear cells infiltration. Hyparrhenia hirta supplementation showed a remarkable amelioration of the abnormalities cited above. The results concluded that the treatment with Hyparrhenia hirta had a significant role in protecting the animals from nitrate-induced kidney dysfunction.Keywords: flavonoids, hyparrhenia hirta, kidney, nitrate toxicity, oxidative stress, rat
Procedia PDF Downloads 445667 Insights into Insect Vectors: Liberibacter Interactions
Authors: Murad Ghanim
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The citrus greening disease, also known as Huanglongbing, caused by the phloem-limited bacterium Candidatus Liberibacter asiaticus (CLas) has resulted in tremendous losses and the death of millions of citrus trees worldwide. CLas is transmitted by the Asian citrus psyllid (ACP) Diaphorina citri. The closely-related bacterium Candidatus Liberibacter solanacearum (CLso), which is associated with vegetative disorders in carrots and the zebra chips disease in potatoes, is transmitted by other psyllid species including Bactericera trigonica in carrots and B. ckockerelli in potatoes. Chemical sprays are currently the prevailing method for managing these diseases for limiting psyllid populations; however, they are limited in their effectiveness. A promising approach to prevent the transmission of these pathogens is to interfere with the vector-pathogen interactions, but our understanding of these processes is very limited. CLas induces changes in the nuclear architecture in the midgut of ACP and activates programmed cell death (apoptosis) in this organ. Strikingly, CLso displayed an opposite effect in the gut of B. trigonica, showing limited apoptosis, but widespread necrosis. Electron and fluorescent microscopy further showed that CLas induced the formation of Endoplasmic reticulum (ER) inclusion- and replication-like bodies, in which it increases and multiplies. ER involvement in bacterial replication is hypothesized to be the first stage of an immune response leading to the apoptotic and necrotic responses. ER exploitation and the subsequent events that lead to these cellular and stress responses might activate a cascade of molecular responses ending up with apoptosis and necrosis. Understanding the molecular interactions that underlay the necrotic/apoptotic responses to the bacteria will increase our knowledge of ACP-CLas, and BT-CLso interactions, and will set the foundation for developing novel, and efficient strategies to disturb these interactions and inhibit the transmission.Keywords: Liberibacter, psyllid, transmission, apoptosis, necrosis
Procedia PDF Downloads 146666 Analysis of Rock Cutting Progress with a New Axe-Shaped PDC Cutter to Improve PDC Bit Performance in Elastoplastic Formation
Authors: Fangyuan Shao, Wei Liu, Deli Gao
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Polycrystalline diamond compact (PDC) bits have occupied a large market of unconventional oil and gas drilling. The application of PDC bits benefits from the efficient rock breaking of PDC cutters. In response to increasingly complex formations, many shaped cutters have been invited, but many of them have not been solved by the mechanism of rock breaking. In this paper, two kinds of PDC cutters: a new axe-shaped (NAS) cutter and cylindrical cutter (benchmark) were studied by laboratory experiments. NAS cutter is obtained by optimizing two sides of axe-shaped cutter with curved surfaces. All the cutters were put on a vertical turret lathe (VTL) in the laboratory for cutting tests. According to the cutting distance, the VTL tests can be divided into two modes: single-turn rotary cutting and continuous cutting. The cutting depth of cutting (DOC) was set at 1.0 mm and 2.0 mm in the former mode. The later mode includes a dry VTL test for thermal stability and a wet VTL test for wear resistance. Load cell and 3D optical profiler were used to obtain the value of cutting forces and wear area, respectively. Based on the findings of the single-turn rotary cutting VTL tests, the performance of A NAS cutter was better than the benchmark cutter on elastoplastic material cutting. The cutting forces (normal forces, tangential force, and radial force) and special mechanical energy (MSE) of a NAS cutter were lower than that of the benchmark cutter under the same condition. It meant that a NAS cutter was more efficient on elastoplastic material breaking. However, the wear resistance of a new axe-shaped cutter was higher than that of a benchmark cutter. The results of the dry VTL test showed that the thermal stability of a NAS cutter was higher than that of a benchmark cutter. The cutting efficiency can be improved by optimizing the geometric structure of the PDC cutter. The change of thermal stability may be caused by the decrease of the contact area between cutter and rock at given DOC. The conclusions of this paper can be used as an important reference for PDC cutters designers.Keywords: axe-shaped cutter, PDC cutter, rotary cutting test, vertical turret lathe
Procedia PDF Downloads 207