Search results for: cell membrane integrity
3863 A Project in the Framework “Nextgenerationeu”: Sustainable Photoelectrochemical Hydrogen Evolution - SERGIO
Authors: Patrizia Frontera, Anastasia Macario, Simona Crispi, Angela Malara, Pierantonio De Luca, Stefano Trocino
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The exploration of solar energy for the photoelectrochemical splitting of water into hydrogen and oxygen has been extensively researched as a means of generating sustainable H₂ fuel. However, despite these efforts, commercialization of this technology has not yet materialized. Presently, the primary impediments to commercialization include low solar-to-hydrogen efficiency (2-3% in PEC with an active area of up to 10-15 cm²), the utilization of costly and critical raw materials (e.g., BiVO₄), and energy losses during the separation of H₂ from O₂ and H₂O vapours in the output stream. The SERGIO partners have identified an advanced approach to fabricate photoelectrode materials, coupled with an appropriate scientific direction to achieve cost-effective solar-driven H₂ production in a tandem photoelectrochemical cell. This project is designed to reach Technology Readiness Level (TRL) 4 by validating the technology in the laboratory using a cell with an active area of up to 10 cm², boasting a solar-to-hydrogen efficiency of 5%, and ensuring acceptable hydrogen purity (99.99%). Our objectives include breakthroughs in cost efficiency, conversion efficiency, and H₂ purity.Keywords: photoelectrolysis, green hydrogen, photoelectrochemical cell, semiconductors
Procedia PDF Downloads 673862 A Stokes Optimal Control Model of Determining Cellular Interaction Forces during Gastrulation
Authors: Yuanhao Gao, Ping Lin, Kees Weijer
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An optimal control system model is proposed for the cell flow in the process of chick embryo gastrulation in this paper. The target is to determine the cellular interaction forces which are hard to measure. This paper will take an approach to investigate the forces with the idea of the inverse problem. By choosing the forces as the control variable and regarding the cell flow as Stokes fluid, an objective functional will be established to match the numerical result of cell velocity with the experimental data. So that the forces could be determined by minimizing the objective functional. The Lagrange multiplier method is utilized to derive the state and adjoint equations consisting the optimal control system, which specifies the first-order necessary conditions. Finite element method is used to discretize and approximate equations. A conjugate gradient algorithm is given for solving the minimum solution of the system and determine the forces.Keywords: optimal control model, Stokes equation, conjugate gradient method, finite element method, chick embryo gastrulation
Procedia PDF Downloads 2593861 Design of Liquid Crystal Based Interface to Study the Interaction of Gram Negative Bacterial Endotoxin with Milk Protein Lactoferrin
Authors: Dibyendu Das, Santanu Kumar Pal
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Milk protein lactoferrin (Lf) exhibits potent antibacterial activity due to its interaction with Gram-negative bacterial cell membrane component, lipopolysaccharide (LPS). This paper represents fabrication of new Liquid crystals (LCs) based biosensors to explore the interaction between Lf and LPS. LPS self-assembled at aqueous/LCs interface and orients interfacial nematic 4-cyano-4’- pentylbiphenyl (5CB) LCs in a homeotropic fashion (exhibiting dark optical image under polarized optical microscope). Interestingly, on the exposure of Lf on LPS decorated aqueous/LCs interface, an optical image of LCs changed from dark to bright indicating an ordering alteration of interfacial LCs from homeotropic to tilted/planar state. The ordering transition reflects strong binding between Lf and interfacial LPS that, in turn, perturbs the orientation of LCs. With the help of epifluorescence microscopy, we further affirmed the interfacial LPS-Lf binding event by imaging the presence of FITC tagged Lf at the LPS laden aqueous/LCs interface. Finally, we have investigated the conformational behavior of Lf in solution as well as in the presence of LPS using Circular Dichroism (CD) spectroscopy and further reconfirmed with Vibrational Circular Dichroism (VCD) spectroscopy where we found that Lf undergoes alpha-helix to random coil-like structure in the presence of LPS. As a whole the entire results described in this paper establish a robust approach to envisage the interaction between LPS and Lf through the ordering transitions of LCs at aqueous/LCs interface.Keywords: endotoxin, interface, lactoferrin, lipopolysaccharide
Procedia PDF Downloads 2663860 Five-Phase Induction Motor Drive System Driven by Five-Phase Packed U Cell Inverter: Its Modeling and Performance Evaluation
Authors: Mohd Tariq
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The three phase system drives produce the problem of more torque pulsations and harmonics. This issue prevents the smooth operation of the drives and it also induces the amount of heat generated thus resulting in an increase in power loss. Higher phase system offers smooth operation of the machines with greater power capacity. Five phase variable-speed induction motor drives are commonly used in various industrial and commercial applications like tractions, electrical vehicles, ship propulsions and conveyor belt drive system. In this work, a comparative analysis of the different modulation schemes applied on the five-level five-phase Packed U Cell (PUC) inverter fed induction motor drives is presented. The performance of the inverter is greatly affected with the modulation schemes applied. The system is modeled, designed, and implemented in MATLAB®/Simulink environment. Experimental validation is done for the prototype of single phase, whereas five phase experimental validation is proposed in the future works.Keywords: Packed U-Cell (PUC) inverter, five-phase system, pulse width modulation (PWM), induction motor (IM)
Procedia PDF Downloads 1833859 Development and Characterisation of a Microbioreactor 'Cassette' for Cell Culture Applications
Authors: Nelson Barrientos, Matthew J. Davies, Marco C. Marques, Darren N. Nesbeth, Gary J. Lye, Nicolas Szita
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Microbioreactor technology is making important advances towards its application in cell culture and bioprocess development. In particular, the technology promises flexible and controllable devices capable to perform parallelised experimentation at low cost. Currently, state of the art methods (e.g. optical sensors) allow the accurate monitoring of the microbioreactor operation. In addition, the laminar flow regime encountered in these devices allows more predictive fluid dynamics modelling, improving the control over the soluble, physical and mechanical environment of the cells. This work describes the development and characterisation of a novel microbioreactor cassette system (microbioreactor volume is 150 μL. The volumetric oxygen transfer coefficient (KLa) and mixing time have been characterised to be between 25 to 113 h-1 and 0.5 and 0.1 s, respectively. In addition, the Residence time distribution (RTD) analysis confirms that the reactor operates at well mixed conditions. Finally, Staphylococcus carnosus TM300 growth is demonstrated via batch culture experiments. Future work consists in expanding the optics of the microbioreactor design to include the monitoring of variables such as fluorescent protein expression, among others.Keywords: microbioreactor, cell-culture, fermentation, microfluidics
Procedia PDF Downloads 4163858 Role of Interleukin 6 on Cell Differentiations in Stem Cells Isolated from Human Exfoliated Deciduous Teeth
Authors: Nunthawan Nowwarote, Waleerat Sukarawan, Prasit Pavasant, Thanaphum Osathanon
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Interleukin 6 (IL-6) is a multifunctional cytokine, regulating various biological responses in several tissues. A Recent study shows that IL-6 plays a role in stemness maintenance in stem cells isolated from human exfoliated deciduous teeth (SHEDs). However, the role of IL-6 on cell differentiation in SHEDs remains unknown. The present study investigated the effect of IL-6 on SHEDs differentiation. Cells were isolated from dental pulp tissues of human deciduous teeth. Flow cytometry was used to determined mesenchymal stem cell marker expression, and the multipotential differentiation (osteogenic, adipogenic and neurogenic lineage ) was also determined. The mRNA was determined using real-time quantitative polymerase chain reaction, and the phenotypes were confirmed by chemical and immunofluorescence staining. Results demonstrated that SHEDs expressed CD44, CD73, CD90, CD105 but not CD45. Further, the up-regulation of osteogenic, adipogenic and neurogenic marker genes was observed upon maintaining cells in osteogenic, adipogenic and neurogenic induction medium, respectively. The addition of IL-6 induced osteogenic by up-regulated osteogenic marker gene also increased in vitro mineralization. Under neurogenic medium supplement with IL-6, up-regulated neurogenic marker. Whereas, an addition of IL-6 attenuated adipogenic differentiation by SHEDs. In conclusion, this evidence implies that IL-6 may participate in cells differentiation ability of SHEDs.Keywords: SHEDs, IL-6, cell differentiations, dental pulp
Procedia PDF Downloads 1803857 Phenotype of Cutaneous Squamous Cell Carcinoma in a Brazilian City with a Tropical Climate
Authors: Julia V. F. Cortes, Maria E. V. Amarante, Carolina L. Cerdeira, Roberta B. V. Silva
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Nonmelanoma skin cancer is more commonly diagnosed than all other malignancies combined. In that group, cutaneous squamous cell carcinoma stands out for having the highest probability of metastasis and recurrence after treatment, in addition to being the second most prevalent form of skin cancer. Its main risk factors include exposure to carcinogens, such as ultraviolet radiation related to sunlight exposure, smoking, alcohol consumption, and human papillomavirus (HPV) infection. Considering the increased risk of skin cancer in the Brazilian population, caused by the high incidence of solar radiation, and the importance of identifying risk phenotypes for the accomplishment of public health actions, an epidemiological study was conducted in a city with a tropical climate located in southeastern Brazil, aiming to identify the target population and assist in primary and secondary prevention. This study describes the profile of patients with cutaneous squamous cell cancer, correlating the variables, sex, age, and differentiation. The study used as primary data source the results of anatomopathological exams delivered from January 2015 to December 2019 for patients registered at one pathology service, which analyzes the results of biopsies, Thus, 66 patients with cutaneous squamous cell carcinoma were analyzed. The most affected age group was 60 years or older (78.79%), emphasizing that moderately differentiated (79.49%) and well-differentiated forms (66.67%) are prevalent in this age group, resulting in a difference of 12.82 percentage points between them. In addition, the predominant sex was male (58%), and it was found that half of the women and 65.79% of men had a moderately differentiated type, whereas the well-differentiated type was slightly more frequent in women. It is worth noting that the moderately differentiated subtype has a 59.20% prevalence among all cases. Thus, it was concluded that the most affected age group was 60 years or older and that men were more affected. As for the subtype, the moderately differentiated one, which is recognized for presenting the second-highest risk for metastasis, was prevalent in this study, affecting 6.6% more men and predominating in the elderly.Keywords: cutaneous squamous cell carcinoma, epidemiology, skin cancer, spinal cell cancer
Procedia PDF Downloads 1173856 TNF Modulation of Cancer Stem Cells in Renal Clear Cell Carcinoma
Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley
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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133
Procedia PDF Downloads 2633855 Constitutive Flo1p Expression on Strains Bearing Deletions in Genes Involved in Cell Wall Biogenesis
Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender
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The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.Keywords: glycoproteins, genetic engineering, flocculation, over-expression
Procedia PDF Downloads 4153854 Investigation of Permeate Flux through DCMD Module by Inserting S-Ribs Carbon-Fiber Promoters with Ascending and Descending Hydraulic Diameters
Authors: Chii-Dong Ho, Jian-Har Chen
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The decline in permeate flux across membrane modules is attributed to the increase in temperature polarization resistance in flat-plate Direct Contact Membrane Distillation (DCMD) modules for pure water productivity. Researchers have discovered that this effect can be diminished by embedding turbulence promoters, which augment turbulence intensity at the cost of increased power consumption, thereby improving vapor permeate flux. The device performance of DCMD modules for permeate flux was further enhanced by shrinking the hydraulic diameters of inserted S-ribs carbon-fiber promoters as well as considering the energy consumption increment. The mass-balance formulation, based on the resistance-in-series model by energy conservation in one-dimensional governing equations, was developed theoretically and conducted experimentally on a flat-plate polytetrafluoroethylene/polypropylene (PTFE/PP) membrane module to predict permeate flux and temperature distributions. The ratio of permeate flux enhancement to energy consumption increment, as referred to an assessment on economic viewpoint and technical feasibilities, was calculated to determine the suitable design parameters for DCMD operations with the insertion of S-ribs carbon-fiber turbulence promoters. An economic analysis was also performed, weighing both permeate flux improvement and energy consumption increment on modules with promoter-filled channels by different array configurations and various hydraulic diameters of turbulence promoters. Results showed that the ratio of permeate flux improvement to energy consumption increment in descending hydraulic-diameter modules is higher than in uniform hydraulic-diameter modules. The fabrication details of the DCMD module filaments implementing the S-ribs carbon-fiber filaments and the schematic configuration of the flat-plate DCMD experimental setup with presenting acrylic plates as external walls were demonstrated in the present study. The S-ribs carbon fibers perform as turbulence promoters incorporated into the artificial hot saline feed stream, which was prepared by adding inorganic salts (NaCl) to distilled water. Theoretical predictions and experimental results exhibited a great accomplishment to considerably achieve permeate flux enhancement, such as the new design of the DCMD module with inserting S-ribs carbon-fiber promoters. Additionally, the Nusselt number for the water vapor transferring membrane module with inserted S-ribs carbon-fiber promoters was generalized into a simplified expression to predict the heat transfer coefficient and permeate flux as well.Keywords: permeate flux, Nusselt number, DCMD module, temperature polarization, hydraulic diameters
Procedia PDF Downloads 93853 Optimizing Cell Culture Performance in an Ambr15 Microbioreactor Using Dynamic Flux Balance and Computational Fluid Dynamic Modelling
Authors: William Kelly, Sorelle Veigne, Xianhua Li, Zuyi Huang, Shyamsundar Subramanian, Eugene Schaefer
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The ambr15™ bioreactor is a single-use microbioreactor for cell line development and process optimization. The ambr system offers fully automatic liquid handling with the possibility of fed-batch operation and automatic control of pH and oxygen delivery. With operating conditions for large scale biopharmaceutical production properly scaled down, micro bioreactors such as the ambr15™ can potentially be used to predict the effect of process changes such as modified media or different cell lines. In this study, gassing rates and dilution rates were varied for a semi-continuous cell culture system in the ambr15™ bioreactor. The corresponding changes to metabolite production and consumption, as well as cell growth rate and therapeutic protein production were measured. Conditions were identified in the ambr15™ bioreactor that produced metabolic shifts and specific metabolic and protein production rates also seen in the corresponding larger (5 liter) scale perfusion process. A Dynamic Flux Balance model was employed to understand and predict the metabolic changes observed. The DFB model-predicted trends observed experimentally, including lower specific glucose consumption when CO₂ was maintained at higher levels (i.e. 100 mm Hg) in the broth. A Computational Fluid Dynamic (CFD) model of the ambr15™ was also developed, to understand transfer of O₂ and CO₂ to the liquid. This CFD model predicted gas-liquid flow in the bioreactor using the ANSYS software. The two-phase flow equations were solved via an Eulerian method, with population balance equations tracking the size of the gas bubbles resulting from breakage and coalescence. Reasonable results were obtained in that the Carbon Dioxide mass transfer coefficient (kLa) and the air hold up increased with higher gas flow rate. Volume-averaged kLa values at 500 RPM increased as the gas flow rate was doubled and matched experimentally determined values. These results form a solid basis for optimizing the ambr15™, using both CFD and FBA modelling approaches together, for use in microscale simulations of larger scale cell culture processes.Keywords: cell culture, computational fluid dynamics, dynamic flux balance analysis, microbioreactor
Procedia PDF Downloads 2823852 Protective Role of CoQ10 or L-Carnitine on the Integrity of the Myocardium in Doxorubicin Induced Toxicity
Authors: Gehan A. Hegazy, Hesham N. Mustafa, Sally A. El Awdan, Marawan AbdelBaset
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Doxorubicin (DOX) is a chemotherapeutic agent used for the treatment of different cancers and its clinical usage is hindered by the oxidative injury-related cardiotoxicity. This work aims to declare if the harmful effects of DOX on the heart can be alleviated with the use of Coenzyme Q10 (CoQ10) or L-carnitine. The study was performed on seventy-two female Wistar albino rats divided into six groups, 12 animals each: Control group; DOX group (10 mg/kg); CoQ10 group (200 mg/kg); L-carnitine group (100 mg/kg); DOX + CoQ10 group; DOX + L-carnitine group. CoQ10 and L-carnitine treatment orally started five days before a single dose of 10 mg/kg DOX that injected intraperitoneally (IP) then the treatment continued for ten days. At the end of the study, serum biochemical parameters of cardiac damage, oxidative stress indices, and histopathological changes were investigated. CoQ10 or L-carnitine showed noticeable effects in improving cardiac functions evidenced reducing serum enzymes as serum interleukin-1 beta (IL-1), tumor necrosis factor alpha (TNF-), leptin, lactate dehydrogenase (LDH), Cardiotrophin-1, Troponin-I and Troponin-T. Also, alleviate oxidative stress, decrease of cardiac Malondialdehyde (MDA), Nitric oxide (NO) and restoring cardiac reduced glutathione levels to normal levels. Both corrected the cardiac alterations histologically and ultrastructurally. With visible improvements in -SMA, vimentin and eNOS immunohistochemical markers. CoQ10 or L-carnitine supplementation improves the functional and structural integrity of the myocardium.Keywords: CoQ10, doxorubicin, L-Carnitine, cardiotoxicity
Procedia PDF Downloads 1703851 Electricity Production Enhancement in a Constructed Microbial Fuel Cell MFC Using Iron Nanoparticles
Authors: Khaoula Bensaida, Osama Eljamal
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The electrical energy generation through Microbial Fuel Cells (MFCs) using microorganisms is a renewable and sustainable approach. It creates truly an efficient technology for power production and wastewater treatment. MFC is an electrochemical device which turns wastewater into electricity. The most important part of MFC is microbes. Nano zero-valent Iron NZVI technique was successfully applied in degrading the chemical pollutants and cleaning wastewater. However, the use of NZVI for enhancing the current production is still not confirmed yet. This study aims to confirm the effect of these particles on the current generation by using MFC. A constructed microbial fuel cell, which utilizes domestic wastewater, has been considered for wastewater treatment and bio-electricity generation. The two electrodes were connected to an external resistor (200 ohms). Experiments were conducted in two steps. First, the MFC was constructed without adding NZVI particles (Control) while at a second step, nanoparticles were added with a concentration of 50mg/L. After 20 hours, the measured voltage increased to 5 and 8mV, respectively. To conclude, the use of zero-valent iron in an MFC system can increase electricity generation.Keywords: bacterial growth, electricity generation, microbial fuel cell MFC, nano zero-valent iron NZVI.
Procedia PDF Downloads 1443850 Separation of Copper(II) and Iron(III) by Solvent Extraction and Membrane Processes with Ionic Liquids as Carriers
Authors: Beata Pospiech
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Separation of metal ions from aqueous solutions is important as well as difficult process in hydrometallurgical technology. This process is necessary for obtaining of clean metals. Solvent extraction and membrane processes are well known as separation methods. Recently, ionic liquids (ILs) are very often applied and studied as extractants and carriers of metal ions from aqueous solutions due to their good extractability properties for various metals. This work discusses a method to separate copper(II) and iron(III) from hydrochloric acid solutions by solvent extraction and transport across polymer inclusion membranes (PIM) with the selected ionic liquids as extractants/ion carriers. Cyphos IL 101 (trihexyl(tetradecyl)phosphonium chloride), Cyphos IL 104 (trihexyl(tetradecyl)phosphonium bis(2,4,4 trimethylpentyl)phosphi-nate), trioctylmethylammonium thiosalicylate [A336][TS] and trihexyl(tetradecyl)phosphonium thiosalicylate [PR4][TS] were used for the investigations. Effect of different parameters such as hydrochloric acid concentration in aqueous phase on iron(III) and copper(II) extraction has been investigated. Cellulose triacetate membranes with the selected ionic liquids as carriers have been prepared and applied for transport of iron(IIII) and copper(II) from hydrochloric acid solutions.Keywords: copper, iron, ionic liquids, solvent extraction
Procedia PDF Downloads 2793849 Single Cell and Spatial Transcriptomics: A Beginners Viewpoint from the Conceptual Pipeline
Authors: Leo Nnamdi Ozurumba-Dwight
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Messenger ribooxynucleic acid (mRNA) molecules are compositional, protein-based. These proteins, encoding mRNA molecules (which collectively connote the transcriptome), when analyzed by RNA sequencing (RNAseq), unveils the nature of gene expression in the RNA. The obtained gene expression provides clues of cellular traits and their dynamics in presentations. These can be studied in relation to function and responses. RNAseq is a practical concept in Genomics as it enables detection and quantitative analysis of mRNA molecules. Single cell and spatial transcriptomics both present varying avenues for expositions in genomic characteristics of single cells and pooled cells in disease conditions such as cancer, auto-immune diseases, hematopoietic based diseases, among others, from investigated biological tissue samples. Single cell transcriptomics helps conduct a direct assessment of each building unit of tissues (the cell) during diagnosis and molecular gene expressional studies. A typical technique to achieve this is through the use of a single-cell RNA sequencer (scRNAseq), which helps in conducting high throughput genomic expressional studies. However, this technique generates expressional gene data for several cells which lack presentations on the cells’ positional coordinates within the tissue. As science is developmental, the use of complimentary pre-established tissue reference maps using molecular and bioinformatics techniques has innovatively sprung-forth and is now used to resolve this set back to produce both levels of data in one shot of scRNAseq analysis. This is an emerging conceptual approach in methodology for integrative and progressively dependable transcriptomics analysis. This can support in-situ fashioned analysis for better understanding of tissue functional organization, unveil new biomarkers for early-stage detection of diseases, biomarkers for therapeutic targets in drug development, and exposit nature of cell-to-cell interactions. Also, these are vital genomic signatures and characterizations of clinical applications. Over the past decades, RNAseq has generated a wide array of information that is igniting bespoke breakthroughs and innovations in Biomedicine. On the other side, spatial transcriptomics is tissue level based and utilized to study biological specimens having heterogeneous features. It exposits the gross identity of investigated mammalian tissues, which can then be used to study cell differentiation, track cell line trajectory patterns and behavior, and regulatory homeostasis in disease states. Also, it requires referenced positional analysis to make up of genomic signatures that will be sassed from the single cells in the tissue sample. Given these two presented approaches to RNA transcriptomics study in varying quantities of cell lines, with avenues for appropriate resolutions, both approaches have made the study of gene expression from mRNA molecules interesting, progressive, developmental, and helping to tackle health challenges head-on.Keywords: transcriptomics, RNA sequencing, single cell, spatial, gene expression.
Procedia PDF Downloads 1223848 Non-melanoma Nasal Skin Cancer: Literature Review
Authors: Geovanna dos Santos Romeiro, Polintia Rayza Brito da Silva, Luis Henrique Moura, Izadora Moreira Do Amaral, Marília Vitória Pinto Milhomem
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Introduction: The nose is one of the most likely sites for the appearance of malignancy on the face. This can be associated with its unique position of exposure to environmental damage, lack of photoprotection and because it is an area susceptible to greater sun exposure. It is already known that the most common type of nasal tumor is basal cell carcinoma. Squamous cell carcinoma is less common but considerably more aggressive, with a tendency to grow rapidly and metastasize. Nasal skin cancer can have a good prognosis, regardless of the type of treatment chosen, i.e., surgery, radiotherapy or electrodissection. However, tumors that are not diagnosed and treated quickly can be harmful and have a greater chance of metastasizing. When curative surgery is performed, therapies and reconstructive surgical procedures are usually required. Objective: The objective is to review the literature on nasal skin tumors and their types and specific locations. Forty-four articles published in Pubmed related to the location of skin cancer in the specific nasal areas region were analyzed. Twelve were excluded for being prior to the year 2000, three with inconclusive results, and one with unbiased conclusions. Results and Conclusion: Regarding the prevalence of types of nasal tumors, basal cell carcinoma comprises the majority, occurring predominantly in the ala, tip and root; squamous cell carcinoma, on the other hand, is more common in the lateral borders and columella. Even so, 2 articles report that the prevalence of metastasis has a higher incidence in squamous cell carcinomas. All of this points to the importance of early location, including regions that are often overlooked in the examination if the patient is wearing glasses. This topic needs further investigation for a greater correlation between anatomy and clinical-surgical implications.Keywords: skin cancer, melanoma, non-melanoma, surgery
Procedia PDF Downloads 533847 The Transport of Radical Species to Single and Double Strand Breaks in the Liver’s DNA Molecule by a Hybrid Method of Type Monte Carlo - Diffusion Equation
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The therapeutic utility of certain Auger emitters such as iodine-125 depends on their position within the cell nucleus . Or diagnostically, and to maintain as low as possible cell damage, it is preferable to have radionuclide localized outside the cell or at least the core. One solution to this problem is to consider markers capable of conveying anticancer drugs to the tumor site regardless of their location within the human body. The objective of this study is to simulate the impact of a complex such as bleomycin on single and double strand breaks in the DNA molecule. Indeed, this simulation consists of the following transactions: - Construction of BLM -Fe- DNA complex. - Simulation of the electron’s transport from the metastable state excitation of Fe 57 by the Monte Carlo method. - Treatment of chemical reactions in the considered environment by the diffusion equation. For physical, physico-chemical and finally chemical steps, the geometry of the complex is considered as a sphere of 50 nm centered on the binding site , and the mathematical method used is called step by step based on Monte Carlo codes.Keywords: concentration, yield, radical species, bleomycin, excitation, DNA
Procedia PDF Downloads 4573846 Technological Ensuring of the Space Reflector Antennas Manufacturing Process from Carbon Fiber Reinforced Plastics
Authors: Pyi Phyo Maung
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In the study, the calculations of the permeability coefficient, values of the volume and porosity of a unit cell of a woven fabric before and after deformation based on the geometrical parameters are presented. Two types of carbon woven fabric structures were investigated: standard type, which integrated the filament, has a cross sectional shape of a cylinder and spread tow type, which has a rectangular cross sectional shape. The space antennas reflector, which distinctive feature is the presence of the surface of double curvature, is considered as the object of the research. Modeling of the kinetics of the process of impregnation of the reflector for the two types of carbon fabric’s unit cell structures was performed using software RAM-RTM. This work also investigated the influence of the grid angle between warp and welt of the unit cell on the duration of impregnation process. The results showed that decreasing the angle between warp and welt of the unit cell, the decreasing of the permeability values were occurred. Based on the results of calculation samples of the reflectors, their quality was determined. The comparisons of the theoretical and experimental results have been carried out. Comparison of the two textile structures (standard and spread tow) showed that the standard textiles with circular cross section were impregnated faster than spread tows, which have a rectangular cross section.Keywords: vacuum assistant resin infusion, impregnation time, shear angle, reflector and modeling
Procedia PDF Downloads 2733845 Homing of B Cells via Afferent Lymphatics
Authors: Sara Pereira-Nogueira, Tim Worbs, Marc Permanyer-Bosser, Reinhold Förster
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While the entry mechanism of lymphocytes into the lymph node via the blood are well described, it is still largely unknown how cells enter lymph nodes that arrive via afferent lymphatics. In order to address this, our group has established a micro-injection technique in mice through which cells are delivered directly into the lymphatic vessel immediately afferent to the popliteal lymph node. Injected cells can then be tracked via multi-colour fluorescence or 2-photon microscopy, and their localization can be analysed within the popliteal or downstream lymph nodes by immunohistology. Since naïve B cells express the chemokine receptor CXCR5 we intra-lymphatically co-injected B cells derived from wildtype and Cxcr5-deficient mice. While CXCR5 does not play a role in guiding B cells out of the subcapsular sinus, it affects their positioning within the lymph node parenchyma, since CXCR5-deficient B cells are impaired in migrating into the B cell follicle. The knowledge obtained by studying B-cell migration may prove beneficial in clinical settings regarding tumor metastasis or autoimmune diseases.Keywords: afferent lymphatics, B cell migration, chemokine, intra-lymphatic injection
Procedia PDF Downloads 2653844 Precursor Muscle Cell’s Phenotype under Compression in a Biomimetic Mechanical Niche
Authors: Fatemeh Abbasi, Arne Hofemeier, Timo Betz
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Muscle growth and regeneration critically depend on satellite cells (SCs) which are muscle stem cells located between the basal lamina and myofibres. Upon damage, SCs become activated, enter the cell cycle, and give rise to myoblasts that form new myofibres, while a sub-population self-renew and re-populate the muscle stem cell niche. In aged muscle as well as in certain muscle diseases such as muscular dystrophy, some of the SCs lose their regenerative ability. Although it is demonstrated that the chemical composition of SCs quiescent niche is different from the activated niche, the mechanism initially activated in the SCs remains unknown. While extensive research efforts focused on potential chemical activation, no such factor has been identified to the author’s best knowledge. However, it is substantiated that niche mechanics affects SCs behaviors, such as stemness and engraftment. We hypothesize that mechanical stress in the healthy niche (homeostasis) is different from the regenerative niche and that this difference could serve as an early signal activating SCs upon fiber damage. To investigate this hypothesis, we develop a biomimetic system to reconstitute both, the mechanical and the chemical environment of the SC niche. Cells will be confined between two elastic polyacrylamide (PAA) hydrogels with controlled elastic moduli and functionalized surface chemistry. By controlling the distance between the PAA hydrogel surfaces, we vary the compression forces exerted by the substrates on the cells, while the lateral displacement of the upper hydrogel will create controlled shear forces. To establish such a system, a simplified system is presented. We engineered a sandwich-like configuration of two elastic PAA layer with stiffnesses between 1 and 10 kPa and confined a precursor myoblast cell line (C2C12) in between these layers. Our initial observations in this sandwich model indicate that C2C12 cells show different behaviors under mechanical compression if compared to a control one-layer gel without compression. Interestingly, this behavior is stiffness-dependent. While the shape of C2C12 cells in the sandwich consisting of two stiff (10 kPa) layers was much more elongated, showing almost a neuronal phenotype, the cell shape in a sandwich situation consisting of one stiff and one soft (1 kPa) layer was more spherical. Surprisingly, even in proliferation medium and at very low cell density, the sandwich situation stimulated cell differentiation with increased striation and myofibre formation. Such behavior is commonly found for confluent cells in differentiation medium. These results suggest that mechanical changes in stiffness and applied pressure might be a relevant stimulation for changes in muscle cell behavior.Keywords: C2C12 cells, compression, force, satellite cells, skeletal muscle
Procedia PDF Downloads 1243843 Treatment of Greywater at Household by Using Ceramic Tablet Membranes
Authors: Abdelkader T. Ahmed
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Greywater is any wastewater draining from a household including kitchen sinks and bathroom tubs, except toilet wastes. Although this used water may contain grease, food particles, hair, and any number of other impurities, it may still be suitable for reuse after treatment. Greywater reusing serves two purposes including reduction the amount of freshwater needed to supply a household, and reduction the amount of wastewater entering sewer systems. This study aims to investigate and design a simple and cheap unit to treat the greywater in household via using ceramic membranes and reuse it in supplying water for toilet flushing. The study include an experimental program for manufacturing several tablet ceramic membranes from clay and sawdust with three different mixtures. The productivity and efficiency of these ceramic membranes were investigated by chemical and physical tests for greywater before and after filtration through these membranes. Then a treatment unit from this ceramic membrane was designed based on the experimental results of lab tests. Results showed that increase sawdust percent with the mixture increase the flow rate and productivity of treated water but decrease in the same time the water quality. The efficiency of the new ceramic membrane reached 95%. The treatment unit save 0.3 m3/day water for toilet flushing without need to consume them from the fresh water supply network.Keywords: ceramic membranes, filtration, greywater, wastewater treatment
Procedia PDF Downloads 3303842 Effect of the Fluid Temperature on the Crude Oil Fouling in the Heat Exchangers of Algiers Refinery
Authors: Rima Harche, Abdelkader Mouheb
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The Algiers refinery as all the other refineries always suffers from the problem of stopping of the tubes of heat exchanger. For that a study experimental of this phenomenon was undertaken in site on the cell of heat exchangers E101 (E101 CBA and E101 EDF) intended for the heating of the crude before its fractionation, which are exposed to the problem of the fouling on the side tubes exchangers. It is of tube-calenders type with head floating. Each cell is made up of three heat exchangers, laid out in series.Keywords: fouling, fluid temperatue , oil, tubular heat exchanger, fouling resistance, modeling, heat transfer coefficient
Procedia PDF Downloads 4323841 Single Cell Rna Sequencing Operating from Benchside to Bedside: An Interesting Entry into Translational Genomics
Authors: Leo Nnamdi Ozurumba-Dwight
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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq
Procedia PDF Downloads 1763840 Real Time PCR Analysis of microRNA Expression in Oral Cancer
Authors: Karl Kingsley
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Many mechanisms are involved in the control of cellular differentiation and growth, which are often dysregulated in many cancers. Many distinct pathways are involved in these mechanisms of control, including deoxyribonuclease (DNA) methyltransferase and histone deacetylase (HDAC) activation that controls both genetic and epigenetic modifications and micro ribonucleic acid (RNA) expression. Less is known about the expression of DNA methyltransferase (DNMT) and HDAC in oral cancers and the effect on microRNA expression. The primary objective of this study was to evaluate the expression of DNMT and HDAC family members in oral cancer and the concomitant expression of cancer-associated microRNAs. Using commercially available oral cancers, including squamous cell carcinoma (SCC)-4, SCC-9, SCC-15, and SCC-25, RNA was extracted and screened for DNMT, HDAC, and microRNA expression using highly-specific primers and quantitative polymerase chain reaction (qPCR). These data revealed low or absent expression of DNMT-1, which is associated with cellular differentiation but increased expression of DNMT-3a and DNMT-3b in all SCC cell lines compared with normal non-cancerous cell controls. In addition, no expression of HDAC1 and HDAC2 expression was found among the normal, non-cancerous cells but was highly expressed in each of the SCC cell lines examined. Differential expression of oncogenic and cancer-associated microRNAs was also observed among the SCC cell lines, including miR-21, miR-133, miR-149, miR-155, miR-365, and miR-720. These findings also appeared to vary according to observed growth rates among these cells. These data may be the first to demonstrate the expression and association between HDAC and DNMT3 family members among oral cancers. In addition, the differential expression of these epigenetic modifiers may be associated with the expression of specific microRNAs in these cancers, which have not previously been observed to the best of the author's knowledge. In addition, some associations and relationships may exist between the expression of these biomarkers and the rates of growth and proliferation, which may suggest that these expression patterns might represent potentially useful biomarkers to determine tumor aggressiveness and other phenotypic behaviors among oral cancers.Keywords: oral cancer, DNA methyltransferase, histone deacetylase, microRNA
Procedia PDF Downloads 1413839 AAV-Mediated Human Α-Synuclein Expression in a Rat Model of Parkinson's Disease –Further Characterization of PD Phenotype, Fine Motor Functional Effects as Well as Neurochemical and Neuropathological Changes over Time
Authors: R. Pussinen, V. Jankovic, U. Herzberg, M. Cerrada-Gimenez, T. Huhtala, A. Nurmi, T. Ahtoniemi
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Targeted over-expression of human α-synuclein using viral-vector mediated gene delivery into the substantia nigra of rats and non-human primates has been reported to lead to dopaminergic cell loss and the formation of α-synuclein aggregates reminiscent of Lewy bodies. We have previously shown how AAV-mediated expression of α-synuclein is seen in the chronic phenotype of the rats over 16 week follow-up period. In the context of these findings, we attempted to further characterize this long term PD related functional and motor deficits as well as neurochemical and neuropathological changes in AAV-mediated α-synuclein transfection model in rats during chronic follow-up period. Different titers of recombinant AAV expressing human α-synuclein (A53T) were stereotaxically injected unilaterally into substantia nigra of Wistar rats. Rats were allowed to recover for 3 weeks prior to initial baseline behavioral testing with rotational asymmetry test, stepping test and cylinder test. A similar behavioral test battery was applied again at weeks 5, 9,12 and 15. In addition to traditionally used rat PD model tests, MotoRater test system, a high speed kinematic gait performance monitoring was applied during the follow-up period. Evaluation focused on animal gait between groups. Tremor analysis was performed on weeks 9, 12 and 15. In addition to behavioral end-points, neurochemical evaluation of dopamine and its metabolites were evaluated in striatum. Furthermore, integrity of the dopamine active transport (DAT) system was evaluated by using 123I- β-CIT and SPECT/CT imaging on weeks 3, 8 and 12 after AAV- α-synuclein transfection. Histopathology was examined from end-point samples at 3 or 12 weeks after AAV- α-synuclein transfection to evaluate dopaminergic cell viability and microglial (Iba-1) activation status in substantia nigra by using stereological analysis techniques. This study focused on the characterization and validation of previously published AAV- α-synuclein transfection model in rats but with the addition of novel end-points. We present the long term phenotype of AAV- α-synuclein transfected rats with traditionally used behavioral tests but also by using novel fine motor analysis techniques and tremor analysis which provide new insight to unilateral effects of AAV α-synuclein transfection. We also present data about neurochemical and neuropathological end-points for the dopaminergic system in the model and how well they correlate with behavioral phenotype.Keywords: adeno-associated virus, alphasynuclein, animal model, Parkinson’s disease
Procedia PDF Downloads 2953838 Battery Grading Algorithm in 2nd-Life Repurposing LI-Ion Battery System
Authors: Ya L. V., Benjamin Ong Wei Lin, Wanli Niu, Benjamin Seah Chin Tat
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This article introduces a methodology that improves reliability and cyclability of 2nd-life Li-ion battery system repurposed as an energy storage system (ESS). Most of the 2nd-life retired battery systems in the market have module/pack-level state-of-health (SOH) indicator, which is utilized for guiding appropriate depth-of-discharge (DOD) in the application of ESS. Due to the lack of cell-level SOH indication, the different degrading behaviors among various cells cannot be identified upon reaching retired status; in the end, considering end-of-life (EOL) loss and pack-level DOD, the repurposed ESS has to be oversized by > 1.5 times to complement the application requirement of reliability and cyclability. This proposed battery grading algorithm, using non-invasive methodology, is able to detect outlier cells based on historical voltage data and calculate cell-level historical maximum temperature data using semi-analytic methodology. In this way, the individual battery cell in the 2nd-life battery system can be graded in terms of SOH on basis of the historical voltage fluctuation and estimated historical maximum temperature variation. These grades will have corresponding DOD grades in the application of the repurposed ESS to enhance system reliability and cyclability. In all, this introduced battery grading algorithm is non-invasive, compatible with all kinds of retired Li-ion battery systems which lack of cell-level SOH indication, as well as potentially being embedded into battery management software for preventive maintenance and real-time cyclability optimization.Keywords: battery grading algorithm, 2nd-life repurposing battery system, semi-analytic methodology, reliability and cyclability
Procedia PDF Downloads 2033837 Blockchain Technology for Secure and Transparent Oil and Gas Supply Chain Management
Authors: Gaurav Kumar Sinha
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The oil and gas industry, characterized by its complex and global supply chains, faces significant challenges in ensuring security, transparency, and efficiency. Blockchain technology, with its decentralized and immutable ledger, offers a transformative solution to these issues. This paper explores the application of blockchain technology in the oil and gas supply chain, highlighting its potential to enhance data security, improve transparency, and streamline operations. By leveraging smart contracts, blockchain can automate and secure transactions, reducing the risk of fraud and errors. Additionally, the integration of blockchain with IoT devices enables real-time tracking and monitoring of assets, ensuring data accuracy and integrity throughout the supply chain. Case studies and pilot projects within the industry demonstrate the practical benefits and challenges of implementing blockchain solutions. The findings suggest that blockchain technology can significantly improve trust and collaboration among supply chain participants, ultimately leading to more efficient and resilient operations. This study provides valuable insights for industry stakeholders considering the adoption of blockchain technology to address their supply chain management challenges.Keywords: blockchain technology, oil and gas supply chain, data security, transparency, smart contracts, IoT integration, real-time tracking, asset monitoring, fraud reduction, supply chain efficiency, data integrity, case studies, industry implementation, trust, collaboration.
Procedia PDF Downloads 373836 Performance Assessment of Recycled Alum Sludge in the Treatment of Textile Industry Effluent in South Africa
Authors: Tony Ngoy Mbodi, Christophe Muanda
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Textile industry is considered as one of the most polluting sectors in terms of effluent volume of discharge and wastewater composition, such as dye, which represents an environmental hazard when discharged without any proper treatment. A study was conducted to investigate the capability of the use of recycled alum sludge (RAS) as an alternative treatment for the reduction of colour, chemical oxygen demand (COD), total dissolved solids (TDS) and pH adjustment from dye based synthetic textile industry wastewater. The coagulation/flocculation process was studied for coagulants of Alum:RAS ratio of, 1:1, 2:1, 1:2 and 0:1. Experiments on treating the synthetic wastewater using membrane filtration and adsorption with corn cobs were also conducted. Results from the coagulation experiment were compared to those from adsorption with corn cobs and membrane filtration experiments conducted on the same synthetic wastewater. The results of the RAS experiments were also evaluated against standard guidelines for industrial effluents treated for discharge purposes in order to establish its level of compliance. Based on current results, it can be concluded that reusing the alum sludge as a low-cost material pretreatment method into the coagulation/flocculation process can offer some advantages such as high removal efficiency for disperse dye and economic savings on overall treatment of the industry wastewater.Keywords: alum, coagulation/flocculation, dye, recycled alum sludge, textile wastewater
Procedia PDF Downloads 3533835 Sustainable Production of Pharmaceutical Compounds Using Plant Cell Culture
Authors: David A. Ullisch, Yantree D. Sankar-Thomas, Stefan Wilke, Thomas Selge, Matthias Pump, Thomas Leibold, Kai Schütte, Gilbert Gorr
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Plants have been considered as a source of natural substances for ages. Secondary metabolites from plants are utilized especially in medical applications but are more and more interesting as cosmetical ingredients and in the field of nutraceuticals. However, supply of compounds from natural harvest can be limited by numerous factors i.e. endangered species, low product content, climate impacts and cost intensive extraction. Especially in the pharmaceutical industry the ability to provide sufficient amounts of product and high quality are additional requirements which in some cases are difficult to fulfill by plant harvest. Whereas in many cases the complexity of secondary metabolites precludes chemical synthesis on a reasonable commercial basis, plant cells contain the biosynthetic pathway – a natural chemical factory – for a given compound. A promising approach for the sustainable production of natural products can be plant cell fermentation (PCF®). A thoroughly accomplished development process comprises the identification of a high producing cell line, optimization of growth and production conditions, the development of a robust and reliable production process and its scale-up. In order to address persistent, long lasting production, development of cryopreservation protocols and generation of working cell banks is another important requirement to be considered. So far the most prominent example using a PCF® process is the production of the anticancer compound paclitaxel. To demonstrate the power of plant suspension cultures here we present three case studies: 1) For more than 17 years Phyton produces paclitaxel at industrial scale i.e. up to 75,000 L in scale. With 60 g/kg dw this fully controlled process which is applied according to GMP results in outstanding high yields. 2) Thapsigargin is another anticancer compound which is currently isolated from seeds of Thapsia garganica. Thapsigargin is a powerful cytotoxin – a SERCA inhibitor – and the precursor for the derivative ADT, the key ingredient of the investigational prodrug Mipsagargin (G-202) which is in several clinical trials. Phyton successfully generated plant cell lines capable to express this compound. Here we present data about the screening for high producing cell lines. 3) The third case study covers ingenol-3-mebutate. This compound is found in the milky sap of the intact plants of the Euphorbiacae family at very low concentrations. Ingenol-3-mebutate is used in Picato® which is approved against actinic keratosis. Generation of cell lines expressing significant amounts of ingenol-3-mebutate is another example underlining the strength of plant cell culture. The authors gratefully acknowledge Inspyr Therapeutics for funding.Keywords: Ingenol-3-mebutate, plant cell culture, sustainability, thapsigargin
Procedia PDF Downloads 2513834 ATR-IR Study of the Mechanism of Aluminum Chloride Induced Alzheimer Disease - Curative and Protective Effect of Lepidium sativum Water Extract on Hippocampus Rats Brain Tissue
Authors: Maha J. Balgoon, Gehan A. Raouf, Safaa Y. Qusti, Soad S. Ali
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The main cause of Alzheimer disease (AD) was believed to be mainly due to the accumulation of free radicals owing to oxidative stress (OS) in brain tissue. The mechanism of the neurotoxicity of Aluminum chloride (AlCl3) induced AD in hippocampus Albino wister rat brain tissue, the curative & the protective effects of Lipidium sativum group (LS) water extract were assessed after 8 weeks by attenuated total reflection spectroscopy ATR-IR and histologically by light microscope. ATR-IR results revealed that the membrane phospholipid undergo free radical attacks, mediated by AlCl3, primary affects the polyunsaturated fatty acids indicated by the increased of the olefinic -C=CH sub-band area around 3012 cm-1 from the curve fitting analysis. The narrowing in the half band width(HBW) of the sνCH2 sub-band around 2852 cm-1 due to Al intoxication indicates the presence of trans form fatty acids rather than gauch rotomer. The degradation of hydrocarbon chain to shorter chain length, increasing in membrane fluidity, disorder and decreasing in lipid polarity in AlCl3 group were indicated by the detected changes in certain calculated area ratios compared to the control. Administration of LS was greatly improved these parameters compared to the AlCl3 group. Al influences the Aβ aggregation and plaque formation, which in turn interferes to and disrupts the membrane structure. The results also showed a marked increase in the β-parallel and antiparallel structure, that characterize the Aβ formation in Al-induced AD hippocampal brain tissue, indicated by the detected increase in both amide I sub-bands around 1674, 1692 cm-1. This drastic increase in Aβ formation was greatly reduced in the curative and protective groups compared to the AlCl3 group and approaches nearly the control values. These results were supported too by the light microscope. AlCl3 group showed significant marked degenerative changes in hippocampal neurons. Most cells appeared small, shrieked and deformed. Interestingly, the administration of LS in curative and protective groups markedly decreases the amount of degenerated cells compared to the non-treated group. Also the intensity of congo red stained cells was decreased. Hippocampal neurons looked more/or less similar to those of control. This study showed a promising therapeutic effect of Lipidium sativum group (LS) on AD rat model that seriously overcome the signs of oxidative stress on membrane lipid and restore the protein misfolding.Keywords: aluminum chloride, alzheimer disease, ATR-IR, Lipidium sativum
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