Search results for: protein matrix
Commenced in January 2007
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Edition: International
Paper Count: 4498

Search results for: protein matrix

58 A Textile-Based Scaffold for Skin Replacements

Authors: Tim Bolle, Franziska Kreimendahl, Thomas Gries, Stefan Jockenhoevel

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The therapeutic treatment of extensive, deep wounds is limited. Autologous split-skin grafts are used as a so-called ‘gold standard’. Most common deficits are the defects at the donor site, the risk of scarring as well as the limited availability and quality of the autologous grafts. The aim of this project is a tissue engineered dermal-epidermal skin replacement to overcome the limitations of the gold standard. A key requirement for the development of such a three-dimensional implant is the formation of a functional capillary-like network inside the implant to ensure a sufficient nutrient and gas supply. Tailored three-dimensional warp knitted spacer fabrics are used to reinforce the mechanically week fibrin gel-based scaffold and further to create a directed in vitro pre-vascularization along the parallel-oriented pile yarns within a co-culture. In this study various three-dimensional warp knitted spacer fabrics were developed in a factorial design to analyze the influence of the machine parameters such as the stitch density and the pattern of the fabric on the scaffold performance and further to determine suitable parameters for a successful fibrin gel-incorporation and a physiological performance of the scaffold. The fabrics were manufactured on a Karl Mayer double-bar raschel machine DR 16 EEC/EAC. A fine machine gauge of E30 was used to ensure a high pile yarn density for sufficient nutrient, gas and waste exchange. In order to ensure a high mechanical stability of the graft, the fabrics were made of biocompatible PVDF yarns. Key parameters such as the pore size, porosity and stress/strain behavior were investigated under standardized, controlled climate conditions. The influence of the input parameters on the mechanical and morphological properties as well as the ability of fibrin gel incorporation into the spacer fabric was analyzed. Subsequently, the pile yarns of the spacer fabrics were colonized with Human Umbilical Vein Endothelial Cells (HUVEC) to analyze the ability of the fabric to further function as a guiding structure for a directed vascularization. The cells were stained with DAPI and investigated using fluorescence microscopy. The analysis revealed that the stitch density and the binding pattern have a strong influence on both the mechanical and morphological properties of the fabric. As expected, the incorporation of the fibrin gel was significantly improved with higher pore sizes and porosities, whereas the mechanical strength decreases. Furthermore, the colonization trials revealed a high cell distribution and density on the pile yarns of the spacer fabrics. For a tailored reinforcing structure, the minimum porosity and pore size needs to be evaluated which still ensures a complete incorporation of the reinforcing structure into the fibrin gel matrix. That will enable a mechanically stable dermal graft with a dense vascular network for a sufficient nutrient and oxygen supply of the cells. The results are promising for subsequent research in the field of reinforcing mechanically weak biological scaffolds and develop functional three-dimensional scaffolds with an oriented pre-vascularization.

Keywords: fibrin-gel, skin replacement, spacer fabric, pre-vascularization

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57 Glucose Uptake Rate of Insulin-Resistant Human Liver Carcinoma Cells (IR/HepG2) by Flavonoids from Enicostema littorale via IR/IRS1/AKT Pathway

Authors: Priyanka Mokashi, Aparna Khanna, Nancy Pandita

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Diabetes mellitus is a chronic metabolic disorder which will be the 7th leading cause of death by 2030. The current line of treatment for the diabetes mellitus is oral antidiabetic drugs (biguanides, sulfonylureas, meglitinides, thiazolidinediones and alpha-glycosidase inhibitors) and insulin therapy depending upon the type 1 or type 2 diabetes mellitus. But, these treatments have their disadvantages, ranging from the developing of resistance to the drugs and adverse effects caused by them. Alternative to these synthetic agents, natural products provides a new insight for the development of more efficient and safe drugs due to their therapeutic values. Enicostema littorale blume (A. Raynal) is a traditional Indian plant belongs to the Gentianaceae family. It is widely distributed in Asia, Africa, and South America. There are few reports on Swrtiamarin, major component of this plant for its antidiabetic activity. However, the antidiabetic activity of flavonoids from E. littorale and their mechanism of action have not yet been elucidated. Flavonoids have a positive relationship with disease prevention and can act on various molecular targets and regulate different signaling pathways in pancreatic β-cells, adipocytes, hepatocytes and skeletal myofibers. They may exert beneficial effects in diabetes by (i) improving hyperglycemia through regulation of glucose metabolism in hepatocytes; (ii) enhancing insulin secretion and reducing apoptosis and promoting proliferation of pancreatic β-cells; (iii) increasing glucose uptake in hepatocytes, skeletal muscle and white adipose tissue (iv) reducing insulin resistance, inflammation and oxidative stress. Therefore, we have isolated four flavonoid rich fractions, Fraction A (FA), Fraction B (FB), Fraction C (FC), Fraction D (FD) from crude alcoholic hot (AH) extract from E. littorale, identified by LC/MS. Total eight flavonoids were identified on the basis of fragmentation pattern. Flavonoid FA showed the presence of swertisin, isovitexin, and saponarin; FB showed genkwanin, quercetin, isovitexin, FC showed apigenin, swertisin, quercetin, 5-O-glucosylswertisin and 5-O-glucosylisoswertisin whereas FD showed the presence of swertisin. Further, these fractions were assessed for their antidiabetic activity on stimulating glucose uptake in insulin-resistant HepG2 cell line model (IR/HepG2). The results showed that FD containing C-glycoside Swertisin has significantly increased the glucose uptake rate of IR/HepG2 cells at the concentration of 10 µg/ml as compared to positive control Metformin (0.5mM) which was determined by glucose oxidase- peroxidase method. It has been reported that enhancement of glucose uptake of cells occurs due the translocation of Glut4 vesicles to cell membrane through IR/IRS1/AKT pathway. Therefore, we have studied expressions of three genes IRS1, AKT and Glut4 by real-time PCR to evaluate whether they follow the same pathway or not. It was seen that the glucose uptake rate has increased in FD treated IR/HepG2 cells due to the activation of insulin receptor substrate-1 (IRS1) followed by protein kinase B (AKT) through phosphoinositide 3-kinase (PI3K) leading to translocation of Glut 4 vesicles to cell membrane, thereby enhancing glucose uptake and insulin sensitivity of insulin resistant HepG2 cells. Hence, the up-regulation indicated the mechanism of action through which FD (Swertisin) acts as antidiabetic candidate in the treatment of type 2 diabetes mellitus.

Keywords: E. littorale, glucose transporter, glucose uptake rate, insulin resistance

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56 An Integrated Approach to the Carbonate Reservoir Modeling: Case Study of the Eastern Siberia Field

Authors: Yana Snegireva

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Carbonate reservoirs are known for their heterogeneity, resulting from various geological processes such as diagenesis and fracturing. These complexities may cause great challenges in understanding fluid flow behavior and predicting the production performance of naturally fractured reservoirs. The investigation of carbonate reservoirs is crucial, as many petroleum reservoirs are naturally fractured, which can be difficult due to the complexity of their fracture networks. This can lead to geological uncertainties, which are important for global petroleum reserves. The problem outlines the key challenges in carbonate reservoir modeling, including the accurate representation of fractures and their connectivity, as well as capturing the impact of fractures on fluid flow and production. Traditional reservoir modeling techniques often oversimplify fracture networks, leading to inaccurate predictions. Therefore, there is a need for a modern approach that can capture the complexities of carbonate reservoirs and provide reliable predictions for effective reservoir management and production optimization. The modern approach to carbonate reservoir modeling involves the utilization of the hybrid fracture modeling approach, including the discrete fracture network (DFN) method and implicit fracture network, which offer enhanced accuracy and reliability in characterizing complex fracture systems within these reservoirs. This study focuses on the application of the hybrid method in the Nepsko-Botuobinskaya anticline of the Eastern Siberia field, aiming to prove the appropriateness of this method in these geological conditions. The DFN method is adopted to model the fracture network within the carbonate reservoir. This method considers fractures as discrete entities, capturing their geometry, orientation, and connectivity. But the method has significant disadvantages since the number of fractures in the field can be very high. Due to limitations in the amount of main memory, it is very difficult to represent these fractures explicitly. By integrating data from image logs (formation micro imager), core data, and fracture density logs, a discrete fracture network (DFN) model can be constructed to represent fracture characteristics for hydraulically relevant fractures. The results obtained from the DFN modeling approaches provide valuable insights into the East Siberia field's carbonate reservoir behavior. The DFN model accurately captures the fracture system, allowing for a better understanding of fluid flow pathways, connectivity, and potential production zones. The analysis of simulation results enables the identification of zones of increased fracturing and optimization opportunities for reservoir development with the potential application of enhanced oil recovery techniques, which were considered in further simulations on the dual porosity and dual permeability models. This approach considers fractures as separate, interconnected flow paths within the reservoir matrix, allowing for the characterization of dual-porosity media. The case study of the East Siberia field demonstrates the effectiveness of the hybrid model method in accurately representing fracture systems and predicting reservoir behavior. The findings from this study contribute to improved reservoir management and production optimization in carbonate reservoirs with the use of enhanced and improved oil recovery methods.

Keywords: carbonate reservoir, discrete fracture network, fracture modeling, dual porosity, enhanced oil recovery, implicit fracture model, hybrid fracture model

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55 Effects of Heart Rate Variability Biofeedback to Improve Autonomic Nerve Function, Inflammatory Response and Symptom Distress in Patients with Chronic Kidney Disease: A Randomized Control Trial

Authors: Chia-Pei Chen, Yu-Ju Chen, Yu-Juei Hsu

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The prevalence and incidence of end-stage renal disease in Taiwan ranks the highest in the world. According to the statistical survey of the Ministry of Health and Welfare in 2019, kidney disease is the ninth leading cause of death in Taiwan. It leads to autonomic dysfunction, inflammatory response and symptom distress, and further increases the damage to the structure and function of the kidneys, leading to increased demand for renal replacement therapy and risks of cardiovascular disease, which also has medical costs for the society. If we can intervene in a feasible manual to effectively regulate the autonomic nerve function of CKD patients, reduce the inflammatory response and symptom distress. To prolong the progression of the disease, it will be the main goal of caring for CKD patients. This study aims to test the effect of heart rate variability biofeedback (HRVBF) on improving autonomic nerve function (Heart Rate Variability, HRV), inflammatory response (Interleukin-6 [IL-6], C reaction protein [CRP] ), symptom distress (Piper fatigue scale, Pittsburgh Sleep Quality Index [PSQI], and Beck Depression Inventory-II [BDI-II] ) in patients with chronic kidney disease. This study was experimental research, with a convenience sampling. Participants were recruited from the nephrology clinic at a medical center in northern Taiwan. With signed informed consent, participants were randomly assigned to the HRVBF or control group by using the Excel BINOMDIST function. The HRVBF group received four weekly hospital-based HRVBF training, and 8 weeks of home-based self-practice was done with StressEraser. The control group received usual care. We followed all participants for 3 months, in which we repeatedly measured their autonomic nerve function (HRV), inflammatory response (IL-6, CRP), and symptom distress (Piper fatigue scale, PSQI, and BDI-II) on their first day of study participation (baselines), 1 month, and 3 months after the intervention to test the effects of HRVBF. The results were analyzed by SPSS version 23.0 statistical software. The data of demographics, HRV, IL-6, CRP, Piper fatigue scale, PSQI, and BDI-II were analyzed by descriptive statistics. To test for differences between and within groups in all outcome variables, it was used by paired sample t-test, independent sample t-test, Wilcoxon Signed-Rank test and Mann-Whitney U test. Results: Thirty-four patients with chronic kidney disease were enrolled, but three of them were lost to follow-up. The remaining 31 patients completed the study, including 15 in the HRVBF group and 16 in the control group. The characteristics of the two groups were not significantly different. The four-week hospital-based HRVBF training combined with eight-week home-based self-practice can effectively enhance the parasympathetic nerve performance for patients with chronic kidney disease, which may against the disease-related parasympathetic nerve inhibition. In the inflammatory response, IL-6 and CRP in the HRVBF group could not achieve significant improvement when compared with the control group. Self-reported fatigue and depression significantly decreased in the HRVBF group, but they still failed to achieve a significant difference between the two groups. HRVBF has no significant effect on improving the sleep quality for CKD patients.

Keywords: heart rate variability biofeedback, autonomic nerve function, inflammatory response, symptom distress, chronic kidney disease

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54 Electroactive Ferrocenyl Dendrimers as Transducers for Fabrication of Label-Free Electrochemical Immunosensor

Authors: Sudeshna Chandra, Christian Gäbler, Christian Schliebe, Heinrich Lang

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Highly branched dendrimers provide structural homogeneity, controlled composition, comparable size to biomolecules, internal porosity and multiple functional groups for conjugating reactions. Electro-active dendrimers containing multiple redox units have generated great interest in their use as electrode modifiers for development of biosensors. The electron transfer between the redox-active dendrimers and the biomolecules play a key role in developing a biosensor. Ferrocenes have multiple and electrochemically equivalent redox units that can act as electron “pool” in a system. The ferrocenyl-terminated polyamidoamine dendrimer is capable of transferring multiple numbers of electrons under the same applied potential. Therefore, they can be used for dual purposes: one in building a film over the electrode for immunosensors and the other for immobilizing biomolecules for sensing. Electrochemical immunosensor, thus developed, exhibit fast and sensitive analysis, inexpensive and involve no prior sample pre-treatment. Electrochemical amperometric immunosensors are even more promising because they can achieve a very low detection limit with high sensitivity. Detection of the cancer biomarkers at an early stage can provide crucial information for foundational research of life science, clinical diagnosis and prevention of disease. Elevated concentration of biomarkers in body fluid is an early indication of some type of cancerous disease and among all the biomarkers, IgG is the most common and extensively used clinical cancer biomarkers. We present an IgG (=immunoglobulin) electrochemical immunosensor using a newly synthesized redox-active ferrocenyl dendrimer of generation 2 (G2Fc) as glassy carbon electrode material for immobilizing the antibody. The electrochemical performance of the modified electrodes was assessed in both aqueous and non-aqueous media using varying scan rates to elucidate the reaction mechanism. The potential shift was found to be higher in an aqueous electrolyte due to presence of more H-bond which reduced the electrostatic attraction within the amido groups of the dendrimers. The cyclic voltammetric studies of the G2Fc-modified GCE in 0.1 M PBS solution of pH 7.2 showed a pair of well-defined redox peaks. The peak current decreased significantly with the immobilization of the anti-goat IgG. After the immunosensor is blocked with BSA, a further decrease in the peak current was observed due to the attachment of the protein BSA to the immunosensor. A significant decrease in the current signal of the BSA/anti-IgG/G2Fc/GCE was observed upon immobilizing IgG which may be due to the formation of immune-conjugates that blocks the tunneling of mass and electron transfer. The current signal was found to be directly related to the amount of IgG captured on the electrode surface. With increase in the concentration of IgG, there is a formation of an increasing amount of immune-conjugates that decreased the peak current. The incubation time and concentration of the antibody was optimized for better analytical performance of the immunosensor. The developed amperometric immunosensor is sensitive to IgG concentration as low as 2 ng/mL. Tailoring of redox-active dendrimers provides enhanced electroactivity to the system and enlarges the sensor surface for binding the antibodies. It may be assumed that both electron transfer and diffusion contribute to the signal transformation between the dendrimers and the antibody.

Keywords: ferrocenyl dendrimers, electrochemical immunosensors, immunoglobulin, amperometry

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53 Oncolytic Efficacy of Thymidine Kinase-Deleted Vaccinia Virus Strain Tiantan (oncoVV-TT) in Glioma

Authors: Seyedeh Nasim Mirbahari, Taha Azad, Mehdi Totonchi

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Oncolytic viruses, which only replicate in tumor cells, are being extensively studied for their use in cancer therapy. A particular virus known as the vaccinia virus, a member of the poxvirus family, has demonstrated oncolytic abilities glioma. Treating Glioma with traditional methods such as chemotherapy and radiotherapy is quite challenging. Even though oncolytic viruses have shown immense potential in cancer treatment, their effectiveness in glioblastoma treatment is still low. Therefore, there is a need to improve and optimize immunotherapies for better results. In this study, we have designed oncoVV-TT, which can more effectively target tumor cells while minimizing replication in normal cells by replacing the thymidine kinase gene with a luc-p2a-GFP gene expression cassette. Human glioblastoma cell line U251 MG, rat glioblastoma cell line C6, and non-tumor cell line HFF were plated at 105 cells in a 12-well plates in 2 mL of DMEM-F2 medium with 10% FBS added to each well. Then incubated at 37°C. After 16 hours, the cells were treated with oncoVV-TT at an MOI of 0.01, 0.1 and left in the incubator for a further 24, 48, 72 and 96 hours. Viral replication assay, fluorescence imaging and viability tests, including trypan blue and crystal violet, were conducted to evaluate the cytotoxic effect of oncoVV-TT. The finding shows that oncoVV-TT had significantly higher cytotoxic activity and proliferation rates in tumor cells in a dose and time-dependent manner, with the strongest effect observed in U251 MG. To conclude, oncoVV-TT has the potential to be a promising oncolytic virus for cancer treatment, with a more cytotoxic effect in human glioblastoma cells versus rat glioma cells. To assess the effectiveness of vaccinia virus-mediated viral therapy, we have tested U251mg and C6 tumor cell lines taken from human and rat gliomas, respectively. The study evaluated oncoVV-TT's ability to replicate and lyse cells and analyzed the survival rates of the tested cell lines when treated with different doses of oncoVV-TT. Additionally, we compared the sensitivity of human and mouse glioma cell lines to the oncolytic vaccinia virus. All experiments regarding viruses were conducted under biosafety level 2. We engineered a Vaccinia-based oncolytic virus called oncoVV-TT to replicate specifically in tumor cells. To propagate the oncoVV-TT virus, HeLa cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 10 MOI virus was added. After 48 h, cells were harvested by scraping, and viruses were collected by 3 sequential freezing and thawing cycles followed by removal of cell debris by centrifugation (1500 rpm, 5 min). The supernatant was stored at −80 ◦C for the following experiments. To measure the replication of the virus in Hela, cells (5 × 104/well) were plated in 24-well plates and incubated overnight to attach to the bottom of the wells. Subsequently, 5 MOI virus or equal dilution of PBS was added. At the treatment time of 0 h, 24 h, 48 h, 72 h and 96 h, the viral titers were determined under the fluorescence microscope (BZ-X700; Keyence, Osaka, Japan). Fluorescence intensity was quantified using the imagej software according to the manufacturer’s protocol. For the isolation of single-virus clones, HeLa cells seeded in six-well plates (5×105 cells/well). After 24 h (100% confluent), the cells were infected with a 10-fold dilution series of TianTan green fluorescent protein (GFP)virus and incubated for 4 h. To examine the cytotoxic effect of oncoVV-TT virus ofn U251mg and C6 cell, trypan blue and crystal violet assay was used.

Keywords: oncolytic virus, immune therapy, glioma, vaccinia virus

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52 Stent Surface Functionalisation via Plasma Treatment to Promote Fast Endothelialisation

Authors: Irene Carmagnola, Valeria Chiono, Sandra Pacharra, Jochen Salber, Sean McMahon, Chris Lovell, Pooja Basnett, Barbara Lukasiewicz, Ipsita Roy, Xiang Zhang, Gianluca Ciardelli

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Thrombosis and restenosis after stenting procedure can be prevented by promoting fast stent wall endothelialisation. It is well known that surface functionalisation with antifouling molecules combining with extracellular matrix proteins is a promising strategy to design biomimetic surfaces able to promote fast endothelialization. In particular, REDV has gained much attention for the ability to enhance rapid endothelialization due to its specific affinity with endothelial cells (ECs). In this work, a two-step plasma treatment was performed to polymerize a thin layer of acrylic acid, used to subsequently graft PEGylated-REDV and polyethylene glycol (PEG) at different molar ratio with the aim to selectively promote endothelial cell adhesion avoiding platelet activation. PEGylate-REDV was provided by Biomatik and it is formed by 6 PEG monomer repetitions (Chempep Inc.), with an NH2 terminal group. PEG polymers were purchased from Chempep Inc. with two different chain lengths: m-PEG6-NH2 (295.4 Da) with 6 monomer repetitions and m-PEG12-NH2 (559.7 Da) with 12 monomer repetitions. Plasma activation was obtained by operating at 50W power, 5 min of treatment and at an Ar flow rate of 20 sccm. Pure acrylic acid (99%, AAc) vapors were diluted in Ar (flow = 20 sccm) and polymerized by a pulsed plasma discharge applying a discharge RF power of 200 W, a duty cycle of 10% (on time = 10 ms, off time = 90 ms) for 10 min. After plasma treatment, samples were dipped into an 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC)/N-hydroxysuccinimide (NHS) solution (ratio 4:1, pH 5.5) for 1 h at 4°C and subsequently dipped in PEGylate-REDV and PEGylate-REDV:PEG solutions at different molar ratio (100 μg/mL in PBS) for 20 h at room temperature. Surface modification was characterized through physico-chemical analyses and in vitro cell tests. PEGylated-REDV peptide and PEG were successfully bound to the carboxylic groups that are formed on the polymer surface after plasma reaction. FTIR-ATR spectroscopy, X -ray Photoelectron Spectroscopy (XPS) and contact angle measurement gave a clear indication of the presence of the grafted molecules. The use of PEG as a spacer allowed for an increase in wettability of the surface, and the effect was more evident by increasing the amount of PEG. Endothelial cells adhered and spread well on the surfaces functionalized with the REDV sequence. In conclusion, a selective coating able to promote a new endothelial cell layer on polymeric stent surface was developed. In particular, a thin AAc film was polymerised on the polymeric surface in order to expose –COOH groups, and PEGylate-REDV and PEG were successful grafted on the polymeric substrates. The REDV peptide demonstrated to encourage cell adhesion with a consequent, expected improvement of the hemocompatibility of these polymeric surfaces in vivo. Acknowledgements— This work was funded by the European Commission 7th Framework Programme under grant agreement number 604251- ReBioStent (Reinforced Bioresorbable Biomaterials for Therapeutic Drug Eluting Stents). The authors thank all the ReBioStent partners for their support in this work.

Keywords: endothelialisation, plasma treatment, stent, surface functionalisation

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51 Mapping of Urban Micro-Climate in Lyon (France) by Integrating Complementary Predictors at Different Scales into Multiple Linear Regression Models

Authors: Lucille Alonso, Florent Renard

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The characterizations of urban heat island (UHI) and their interactions with climate change and urban climates are the main research and public health issue, due to the increasing urbanization of the population. These solutions require a better knowledge of the UHI and micro-climate in urban areas, by combining measurements and modelling. This study is part of this topic by evaluating microclimatic conditions in dense urban areas in the Lyon Metropolitan Area (France) using a combination of data traditionally used such as topography, but also from LiDAR (Light Detection And Ranging) data, Landsat 8 satellite observation and Sentinel and ground measurements by bike. These bicycle-dependent weather data collections are used to build the database of the variable to be modelled, the air temperature, over Lyon’s hyper-center. This study aims to model the air temperature, measured during 6 mobile campaigns in Lyon in clear weather, using multiple linear regressions based on 33 explanatory variables. They are of various categories such as meteorological parameters from remote sensing, topographic variables, vegetation indices, the presence of water, humidity, bare soil, buildings, radiation, urban morphology or proximity and density to various land uses (water surfaces, vegetation, bare soil, etc.). The acquisition sources are multiple and come from the Landsat 8 and Sentinel satellites, LiDAR points, and cartographic products downloaded from an open data platform in Greater Lyon. Regarding the presence of low, medium, and high vegetation, the presence of buildings and ground, several buffers close to these factors were tested (5, 10, 20, 25, 50, 100, 200 and 500m). The buffers with the best linear correlations with air temperature for ground are 5m around the measurement points, for low and medium vegetation, and for building 50m and for high vegetation is 100m. The explanatory model of the dependent variable is obtained by multiple linear regression of the remaining explanatory variables (Pearson correlation matrix with a |r| < 0.7 and VIF with < 5) by integrating a stepwise sorting algorithm. Moreover, holdout cross-validation is performed, due to its ability to detect over-fitting of multiple regression, although multiple regression provides internal validation and randomization (80% training, 20% testing). Multiple linear regression explained, on average, 72% of the variance for the study days, with an average RMSE of only 0.20°C. The impact on the model of surface temperature in the estimation of air temperature is the most important variable. Other variables are recurrent such as distance to subway stations, distance to water areas, NDVI, digital elevation model, sky view factor, average vegetation density, or building density. Changing urban morphology influences the city's thermal patterns. The thermal atmosphere in dense urban areas can only be analysed on a microscale to be able to consider the local impact of trees, streets, and buildings. There is currently no network of fixed weather stations sufficiently deployed in central Lyon and most major urban areas. Therefore, it is necessary to use mobile measurements, followed by modelling to characterize the city's multiple thermal environments.

Keywords: air temperature, LIDAR, multiple linear regression, surface temperature, urban heat island

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50 Gene Cloning and Expression of Azoreductases from Azo-Degraders Lysinibacillus macrolides and Bacillus coagulans Isolated from Egyptian Industrial Wastewater

Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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49 Extracellular Polymeric Substances (EPS) Attribute to Biofouling of Anaerobic Membrane Bioreactor: Adhesion and Viscoelastic Properties

Authors: Kbrom Mearg Haile

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Introduction: Membrane fouling is the bottleneck for the anaerobic membrane bioreactor (AnMBR) robust continuous operation, primarily caused by the mixed liquor suspended solids (MLSS) characteristics formed by aggregated flocs and a scaffold of microbial self-produced extracellular polymeric substances (EPS), which dictates the flocs integrity. Accordingly, the adhesion of EPS to the membrane surface versus their role in forming firm, elastic, and mechanically stable flocs under the reactor’s hydraulic shear is critical for minimizing interactions between EPS and colloids originating from the MLSS flocs with the membrane. This study aims to gain insight and investigate the effect of MLSS flocs properties, EPS adhesion and viscoelasticity, viscoelastic properties of the sludge, and membrane fouling propensity. Experimental: As a working hypothesis, to alter the aforementioned flocs’ and EPS’s properties, the addition of either coagulant or surfactant was carried out during the AnMBR operation. In the AnMBR, two flat-sheet 300 kDa pore size polyether sulfone (PES) membranes with a total filtration area of 352 cm2 were immersed in the AnMBR system treating municipal wastewater of Midreshet Ben-Gurion village at the Negev highlands, Israel. The system temperature, pH, biogas recirculation, and hydraulic retention time were regulated. TMP fluctuations during a 30-day experiment were recorded under three operating conditions: Baseline (without the addition of coagulating or dispersing agent), coagulant addition (FeCl3), and surfactant addition (sodium dodecyl sulfate). At the end of each experiment, EPS were extracted from the MLSS and from the fouled membrane, characterized for their protein, polysaccharides, and DOC contents, and correlated with the fouling tendency of the submerged UF membrane. The EPS adherence and viscoelastic properties were revealed using QCM-D via the PES-coated gold sensor used as a membrane-mimicking surface providing a detailed real-time EPS adhesion. The associated shifts in the resonance frequency and dissipation at different overtones were further modeled using the Voigt-based viscoelastic model (using Dfind software, Q-Sense Biolin Scientific) in which the thickness, shear modulus, and shear viscosity values of the adsorbed EPS layers on the PES coated sensor were calculated. Results and discussion: The observations obtained from the QCM-D analysis indicate a greater decrease in the frequency shift for the elevated membrane fouling scenarios, likely due to an observed decrease in the calculated shear viscosity and shear modulus of the EPS adsorbed layer, coupled with an increase in EPS layer hydrated thickness and fluidity (ΔD/Δf slopes). Further analysis is being conducted for the three major operating conditions-analyzing their effects on sludge rheology, dewaterability (capillary suction time-CST) and settle ability (SVI). The biofouling layer is further characterized microscopically using a confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM), for analyzing the consistency of the development of the biofouling layer with sludge characteristics, i.e., thicker biofouling layer on the membrane surface when operated with surfactant addition, due to flocs with reduced integrity and availability of EPS/colloids to the membrane. Conversely, a thinner layer when operated with coagulant compared to the baseline experiment, due to elevation in flocs integrity.

Keywords: viscoelasticity, biofouling, viscoelastic, AnMBR, EPS, elocintegrity

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48 Global Evidence on the Seasonality of Enteric Infections, Malnutrition, and Livestock Ownership

Authors: Aishwarya Venkat, Anastasia Marshak, Ryan B. Simpson, Elena N. Naumova

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Livestock ownership is simultaneously linked to improved nutritional status through increased availability of animal-source protein, and increased risk of enteric infections through higher exposure to contaminated water sources. Agrarian and agro-pastoral households, especially those with cattle, goats, and sheep, are highly dependent on seasonally various environmental conditions, which directly impact nutrition and health. This study explores global spatiotemporally explicit evidence regarding the relationship between livestock ownership, enteric infections, and malnutrition. Seasonal and cyclical fluctuations, as well as mediating effects, are further examined to elucidate health and nutrition outcomes of individual and communal livestock ownership. The US Agency for International Development’s Demographic and Health Surveys (DHS) and the United Nations International Children's Emergency Fund’s Multi-Indicator Cluster Surveys (MICS) provide valuable sources of household-level information on anthropometry, asset ownership, and disease outcomes. These data are especially important in data-sparse regions, where surveys may only be conducted in the aftermath of emergencies. Child-level disease history, anthropometry, and household-level asset ownership information have been collected since DHS-V (2003-present) and MICS-III (2005-present). This analysis combines over 15 years of survey data from DHS and MICS to study 2,466,257 children under age five from 82 countries. Subnational (administrative level 1) measures of diarrhea prevalence, mean livestock ownership by type, mean and median anthropometric measures (height for age, weight for age, and weight for height) were investigated. Effects of several environmental, market, community, and household-level determinants were studied. Such covariates included precipitation, temperature, vegetation, the market price of staple cereals and animal source proteins, conflict events, livelihood zones, wealth indices and access to water, sanitation, hygiene, and public health services. Children aged 0 – 6 months, 6 months – 2 years, and 2 – 5 years of age were compared separately. All observations were standardized to interview day of year, and administrative units were harmonized for consistent comparisons over time. Geographically weighted regressions were constructed for each outcome and subnational unit. Preliminary results demonstrate the importance of accounting for seasonality in concurrent assessments of malnutrition and enteric infections. Household assets, including livestock, often determine the intensity of these outcomes. In many regions, livestock ownership affects seasonal fluxes in malnutrition and enteric infections, which are also directly affected by environmental and local factors. Regression analysis demonstrates the spatiotemporal variability in nutrition outcomes due to a variety of causal factors. This analysis presents a synthesis of evidence from global survey data on the interrelationship between enteric infections, malnutrition, and livestock. These results provide a starting point for locally appropriate interventions designed to address this nexus in a timely manner and simultaneously improve health, nutrition, and livelihoods.

Keywords: diarrhea, enteric infections, households, livestock, malnutrition, seasonality

Procedia PDF Downloads 128
47 A Simple Chemical Approach to Regenerating Strength of Thermally Recycled Glass Fibre

Authors: Sairah Bashir, Liu Yang, John Liggat, James Thomason

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Glass fibre is currently used as reinforcement in over 90% of all fibre-reinforced composites produced. The high rigidity and chemical resistance of these composites are required for optimum performance but unfortunately results in poor recyclability; when such materials are no longer fit for purpose, they are frequently deposited in landfill sites. Recycling technologies, for example, thermal treatment, can be employed to address this issue; temperatures typically between 450 and 600 °C are required to allow degradation of the rigid polymeric matrix and subsequent extraction of fibrous reinforcement. However, due to the severe thermal conditions utilised in the recycling procedure, glass fibres become too weak for reprocessing in second-life composite materials. In addition, more stringent legislation is being put in place regarding disposal of composite waste, and so it is becoming increasingly important to develop long-term recycling solutions for such materials. In particular, the development of a cost-effective method to regenerate strength of thermally recycled glass fibres will have a positive environmental effect as a reduced volume of composite material will be destined for landfill. This research study has demonstrated the positive impact of sodium hydroxide (NaOH) and potassium hydroxide (KOH) solution, prepared at relatively mild temperatures and at concentrations of 1.5 M and above, on the strength of heat-treated glass fibres. As a result, alkaline treatments can potentially be implemented to glass fibres that are recycled from composite waste to allow their reuse in second-life materials. The optimisation of the strength recovery process is being conducted by varying certain reaction parameters such as molarity of alkaline solution and treatment time. It is believed that deep V-shaped surface flaws exist commonly on severely damaged fibre surfaces and are effectively removed to form smooth, U-shaped structures following alkaline treatment. Although these surface flaws are believed to be present on glass fibres they have not in fact been observed, however, they have recently been discovered in this research investigation through analytical techniques such as AFM (atomic force microscopy) and SEM (scanning electron microscopy). Reaction conditions such as molarity of alkaline solution affect the degree of etching of the glass fibre surface, and therefore the extent to which fibre strength is recovered. A novel method in determining the etching rate of glass fibres after alkaline treatment has been developed, and the data acquired can be correlated with strength. By varying reaction conditions such as alkaline solution temperature and molarity, the activation energy of the glass etching process and the reaction order can be calculated respectively. The promising results obtained from NaOH and KOH treatments have opened an exciting route to strength regeneration of thermally recycled glass fibres, and the optimisation of the alkaline treatment process is being continued in order to produce recycled fibres with properties that match original glass fibre products. The reuse of such glass filaments indicates that closed-loop recycling of glass fibre reinforced composite (GFRC) waste can be achieved. In fact, the development of a closed-loop recycling process for GFRC waste is already underway in this research study.

Keywords: glass fibers, glass strengthening, glass structure and properties, surface reactions and corrosion

Procedia PDF Downloads 257
46 EGF Serum Level in Diagnosis and Prediction of Mood Disorder in Adolescents and Young Adults

Authors: Monika Dmitrzak-Weglarz, Aleksandra Rajewska-Rager, Maria Skibinska, Natalia Lepczynska, Piotr Sibilski, Joanna Pawlak, Pawel Kapelski, Joanna Hauser

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Epidermal growth factor (EGF) is a well-known neurotrophic factor that involves in neuronal growth and synaptic plasticity. The proteomic research provided in order to identify novel candidate biological markers for mood disorders focused on elevated EGF serum level in patients during depression episode. However, the EGF association with mood disorder spectrum among adolescents and young adults has not been studied extensively. In this study, we aim to investigate the serum levels of EGF in adolescents and young adults during hypo/manic, depressive episodes and in remission compared to healthy control group. In our study, we involved 80 patients aged 12-24 years in 2-year follow-up study with a primary diagnosis of mood disorder spectrum, and 35 healthy volunteers matched by age and gender. Diagnoses were established according to DSM-IV-TR criteria using structured clinical interviews: K-SADS for child and adolescents, and SCID for young adults. Clinical and biological evaluations were made at baseline and euthymic mood (at 3th or 6th month of treatment and after 1 and 2 years). The Young Mania Rating Scale and Hamilton Rating Scale for Depression were used for assessment. The study protocols were approved by the relevant ethics committee. Serum protein concentration was determined by Enzyme-Linked Immunosorbent Assays (ELISA) method. Human EGF (cat. no DY 236) DuoSet ELISA kit was used (R&D Systems). Serum EGF levels were analysed with following variables: age, age under 18 and above 18 years old, sex, family history of affective disorders, drug-free vs. medicated. Shapiro-Wilk test was used to test the normality of the data. The homogeneity of variance was calculated with Levene’s test. EGF levels showed non-normal distribution and the homogeneity of variance was violated. Non-parametric tests: Mann-Whitney U test, Kruskall-Wallis ANOVA, Friedman’s ANOVA, Wilcoxon signed rank test, Spearman correlation coefficient was applied in the analyses The statistical significance level was set at p<0.05. Elevated EGF level at baseline (p=0.001) and at month 24 (p=0.02) was detected in study subjects compared with controls. Increased EGF level in women at month 12 (p=0.02) compared to men in study group have been observed. Using Wilcoxon signed rank test differences in EGF levels were detected: decrease from baseline to month 3 (p=0.014) and increase comparing: month 3 vs. 24 (p=0.013); month 6 vs. 12 (p=0.021) and vs. 24 (p=0.008). EGF level at baseline was negatively correlated with depression and mania occurrence at 24 months. EGF level at 24 months was positively correlated with depression and mania occurrence at 12 months. No other correlations of EGF levels with clinical and demographical variables have been detected. The findings of the present study indicate that EGF serum level is significantly elevated in the study group of patients compared to the controls. We also observed fluctuations in EGF levels during two years of disease observation. EGF seems to be useful as an early marker for prediction of diagnosis, course of illness and treatment response in young patients during first episode od mood disorders, which requires further investigation. Grant was founded by National Science Center in Poland no 2011/03/D/NZ5/06146.

Keywords: biological marker, epidermal growth factor, mood disorders, prediction

Procedia PDF Downloads 190
45 3D Interactions in Under Water Acoustic Simulationseffect of Green Synthesized Metal Nanoparticles on Gene Expression in an In-Vitro Model of Non-alcoholic Steatohepatitis

Authors: Nendouvhada Livhuwani Portia, Nicole Sibuyi, Kwazikwakhe Gabuza, Adewale Fadaka

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Metabolic dysfunction-associated liver disease (MASLD) is a chronic condition characterized by excessive fat accumulation in the liver, distinct from conditions caused by alcohol, viral hepatitis, or medications. MASLD is often linked with metabolic syndrome, including obesity, diabetes, hyperlipidemia, and hypertriglyceridemia. This disease can progress to metabolic dysfunction-associated steatohepatitis (MASH), marked by liver inflammation and scarring, potentially leading to cirrhosis. However, only 43-44% of patients with steatosis develop MASH, and 7-30% of those with MASH progress to cirrhosis. The exact mechanisms underlying MASLD and its progression remain unclear, and there are currently no specific therapeutic strategies for MASLD/MASH. While anti-obesity and anti-diabetic medications can reduce progression, they do not fully treat or reverse the disease. As an alternative, green-synthesized metal nanoparticles (MNPs) are emerging as potential treatments for liver diseases due to their anti-diabetic, anti-inflammatory, and anti-obesity properties with minimal side effects. MNPs like gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) have been shown to improve metabolic processes by lowering blood glucose, body fat, and inflammation. The study aimed to explore the effects of green-synthesized MNPs on gene expression in an in vitro model of MASH using C3A/HepG2 liver cells. The MASH model was created by exposing these cells to free fatty acids (FFAs) followed by lipopolysaccharide (LPS) to induce inflammation. Cell viability was assessed with the Water-Soluble Tetrazolium (WST)-1 assay, and lipid accumulation was measured using the Oil Red O (ORO) assay. Additionally, mitochondrial membrane potential was assessed by the tetramethyl rhodamine, methyl ester (TMRE) assay, and inflammation was measured with an Enzyme-Linked Immunosorbent Assay (ELISA). The study synthesized AuNPs from Carpobrotus edulis fruit (CeF) and avocado seed (AvoSE) and AgNPs from Salvia africana-lutea (SAL) using optimized conditions. The MNPs were characterized by UV-Vis spectrophotometry and Dynamic Light Scattering (DLS). The nanoparticles were tested at various concentrations for their impact on the C3A/HepG2-induced MASH model. Among the MNPs tested, AvoSE-AuNPs showed the most promise. They reduced cell proliferation and intracellular lipid content more effectively than CeFE-AuNPs and SAL-AgNPs. Molecular analysis using real-time polymerase chain reaction revealed that AvoSE-AuNPs could potentially reverse MASH effects by reducing the expression of key pro-inflammatory and metabolic genes, including tumor necrosis factor-alpha (TNF-α), Fas cell surface death receptor (FAS), Peroxisome proliferator-activated receptor (PPAR)-α, PPAR-γ, and Sterol regulatory element-binding protein (SREBPF)-1. Further research is needed to confirm the molecular mechanisms behind the effects of these MNPs and to identify the specific phytochemicals responsible for their synthesis and bioactivities.

Keywords: gold nanoparticles, green nanotechnology, metal nanoparticles, obesity

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44 Analysis of Minimizing Investment Risks in Power and Energy Business Development by Combining Total Quality Management and International Financing Institutions Project Management Tools

Authors: M. Radunovic

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Region of Southeastern Europe has a substantial energy resource potential and is witnessing an increasing rate of power and energy project investments. This comes as a result of countries harmonizing their legal framework and market regulations to conform the ones of European Union, enabling direct private investments. Funding in the power and energy market in this region originates from various resources and investment entities, including commercial and institutional ones. Risk anticipation and assessment is crucial to project success, especially given the long exploitation period of project in power and energy domain, as well as the wide range of stakeholders involved. This paper analyzes the possibility of combined application of tools used in total quality management and international financing institutions for project planning, execution and evaluation, with the goal of anticipating, assessing and minimizing the risks that might occur in the development and execution phase of a power and energy project in the market of southeastern Europe. History of successful project management and investments both in the industry and institutional sector provides sufficient experience, guidance and internationally adopted tools to provide proper project assessment for investments in power and energy. Business environment of southeastern Europe provides immense potential for developing power and engineering projects of various magnitudes, depending on stakeholders’ interest. Diversification on investment sources provides assurance that there is interest and commitment to invest in this market. Global economic and political developments will be intensifying the pace of investments in the upcoming period. The proposed approach accounts for key parameters that contribute to the sustainability and profitability of a project which include technological, educational, social and economic gaps between the southeastern European region and western Europe, market trends in equipment design and production on a global level, environment friendly approach to renewable energy sources as well as conventional power generation systems, and finally the effect of the One Belt One Road Initiative led by People’s Republic of China to the power and energy market of this region in the upcoming period on a long term scale. Analysis will outline the key benefits of the approach as well as the accompanying constraints. Parallel to this it will provide an overview of dominant threats and opportunities in present and future business environment and their influence to the proposed application. Through concrete examples, full potential of this approach will be presented along with necessary improvements that need to be implemented. Number of power and engineering projects being developed in southeastern Europe will be increasing in the upcoming period. Proper risk analysis will lead to minimizing project failures. The proposed successful combination of reliable project planning tools from different investment areas can prove to be beneficial in the future power and engineering investments, and guarantee their sustainability and profitability.

Keywords: capital investments, lean six sigma, logical framework approach, logical framework matrix, one belt one road initiative, project management tools, quality function deployment, Southeastern Europe, total quality management

Procedia PDF Downloads 109
43 Intelligent Materials and Functional Aspects of Shape Memory Alloys

Authors: Osman Adiguzel

Abstract:

Shape-memory alloys are a new class of functional materials with a peculiar property known as shape memory effect. These alloys return to a previously defined shape on heating after deformation in low temperature product phase region and take place in a class of functional materials due to this property. The origin of this phenomenon lies in the fact that the material changes its internal crystalline structure with changing temperature. Shape memory effect is based on martensitic transitions, which govern the remarkable changes in internal crystalline structure of materials. Martensitic transformation, which is a solid state phase transformation, occurs in thermal manner in material on cooling from high temperature parent phase region. This transformation is governed by changes in the crystalline structure of the material. Shape memory alloys cycle between original and deformed shapes in bulk level on heating and cooling, and can be used as a thermal actuator or temperature-sensitive elements due to this property. Martensitic transformations usually occur with the cooperative movement of atoms by means of lattice invariant shears. The ordered parent phase structures turn into twinned structures with this movement in crystallographic manner in thermal induced case. The twinned martensites turn into the twinned or oriented martensite by stressing the material at low temperature martensitic phase condition. The detwinned martensite turns into the parent phase structure on first heating, first cycle, and parent phase structures turn into the twinned and detwinned structures respectively in irreversible and reversible memory cases. On the other hand, shape memory materials are very important and useful in many interdisciplinary fields such as medicine, pharmacy, bioengineering, metallurgy and many engineering fields. The choice of material as well as actuator and sensor to combine it with the host structure is very essential to develop main materials and structures. Copper based alloys exhibit this property in metastable beta-phase region, which has bcc-based structures at high temperature parent phase field, and these structures martensitically turn into layered complex structures with lattice twinning following two ordered reactions on cooling. Martensitic transition occurs as self-accommodated martensite with inhomogeneous shears, lattice invariant shears which occur in two opposite directions, <110 > -type directions on the {110}-type plane of austenite matrix which is basal plane of martensite. This kind of shear can be called as {110}<110> -type mode and gives rise to the formation of layered structures, like 3R, 9R or 18R depending on the stacking sequences on the close-packed planes of the ordered lattice. In the present contribution, x-ray diffraction and transmission electron microscopy (TEM) studies were carried out on two copper based alloys which have the chemical compositions in weight; Cu-26.1%Zn 4%Al and Cu-11%Al-6%Mn. X-ray diffraction profiles and electron diffraction patterns reveal that both alloys exhibit super lattice reflections inherited from parent phase due to the displacive character of martensitic transformation. X-ray diffractograms taken in a long time interval show that locations and intensities of diffraction peaks change with the aging time at room temperature. In particular, some of the successive peak pairs providing a special relation between Miller indices come close each other.

Keywords: Shape memory effect, martensite, twinning, detwinning, self-accommodation, layered structures

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42 Complete Genome Sequence Analysis of Pasteurella multocida Subspecies multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez J. Firdaus Abdullah, Zunita Zakaria, Nurulfiza M. Isa, Yung C. Tan, Wai Y. Yee, Abdul R. Omar

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Pasteurella multocida (PM) is an important veterinary opportunistic pathogen particularly associated with septicemic pasteurellosis, pneumonic pasteurellosis and hemorrhagic septicemia in cattle and buffaloes. P. multocida serotype A has been reported to cause fatal pneumonia and septicemia. Pasteurella multocida subspecies multocida of serotype A Malaysian isolate PMTB2.1 was first isolated from buffaloes died of septicemia. In this study, the genome of P. multocida strain PMTB2.1 was sequenced using third-generation sequencing technology, PacBio RS2 system and analyzed bioinformatically via de novo analysis followed by in-depth analysis based on comparative genomics. Bioinformatics analysis based on de novo assembly of PacBio raw reads generated 3 contigs followed by gap filling of aligned contigs with PCR sequencing, generated a single contiguous circular chromosome with a genomic size of 2,315,138 bp and a GC content of approximately 40.32% (Accession number CP007205). The PMTB2.1 genome comprised of 2,176 protein-coding sequences, 6 rRNA operons and 56 tRNA and 4 ncRNAs sequences. The comparative genome sequence analysis of PMTB2.1 with nine complete genomes which include Actinobacillus pleuropneumoniae, Haemophilus parasuis, Escherichia coli and five P. multocida complete genome sequences including, PM70, PM36950, PMHN06, PM3480, PMHB01 and PMTB2.1 was carried out based on OrthoMCL analysis and Venn diagram. The analysis showed that 282 CDs (13%) are unique to PMTB2.1and 1,125 CDs with orthologs in all. This reflects overall close relationship of these bacteria and supports the classification in the Gamma subdivision of the Proteobacteria. In addition, genomic distance analysis among all nine genomes indicated that PMTB2.1 is closely related with other five Pasteurella species with genomic distance less than 0.13. Synteny analysis shows subtle differences in genetic structures among different P.multocida indicating the dynamics of frequent gene transfer events among different P. multocida strains. However, PM3480 and PM70 exhibited exceptionally large structural variation since they were swine and chicken isolates. Furthermore, genomic structure of PMTB2.1 is more resembling that of PM36950 with a genomic size difference of approximately 34,380 kb (smaller than PM36950) and strain-specific Integrative and Conjugative Elements (ICE) which was found only in PM36950 is absent in PMTB2.1. Meanwhile, two intact prophages sequences of approximately 62 kb were found to be present only in PMTB2.1. One of phage is similar to transposable phage SfMu. The phylogenomic tree was constructed and rooted with E. coli, A. pleuropneumoniae and H. parasuis based on OrthoMCL analysis. The genomes of P. multocida strain PMTB2.1 were clustered with bovine isolates of P. multocida strain PM36950 and PMHB01 and were separated from avian isolate PM70 and swine isolates PM3480 and PMHN06 and are distant from Actinobacillus and Haemophilus. Previous studies based on Single Nucleotide Polymorphism (SNPs) and Multilocus Sequence Typing (MLST) unable to show a clear phylogenetic relatedness between Pasteurella multocida and the different host. In conclusion, this study has provided insight on the genomic structure of PMTB2.1 in terms of potential genes that can function as virulence factors for future study in elucidating the mechanisms behind the ability of the bacteria in causing diseases in susceptible animals.

Keywords: comparative genomics, DNA sequencing, phage, phylogenomics

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41 Impact of Transgenic Adipose Derived Stem Cells in the Healing of Spinal Cord Injury of Dogs

Authors: Imdad Ullah Khan, Yongseok Yoon, Kyeung Uk Choi, Kwang Rae Jo, Namyul Kim, Eunbee Lee, Wan Hee Kim, Oh-Kyeong Kweon

Abstract:

The primary spinal cord injury (SCI) causes mechanical damage to the neurons and blood vessels. It leads to secondary SCI, which activates multiple pathological pathways, which expand neuronal damage at the injury site. It is characterized by vascular disruption, ischemia, excitotoxicity, oxidation, inflammation, and apoptotic cell death. It causes nerve demyelination and disruption of axons, which perpetuate a loss of impulse conduction through the injured spinal cord. It also leads to the production of myelin inhibitory molecules, which with a concomitant formation of an astroglial scar, impede axonal regeneration. The pivotal role regarding the neuronal necrosis is played by oxidation and inflammation. During an early stage of spinal cord injury, there occurs an abundant expression of reactive oxygen species (ROS) due to defective mitochondrial metabolism and abundant migration of phagocytes (macrophages, neutrophils). ROS cause lipid peroxidation of the cell membrane, and cell death. Abundant migration of neutrophils, macrophages, and lymphocytes collectively produce pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1beta (IL-1β), matrix metalloproteinase, superoxide dismutase, and myeloperoxidases which synergize neuronal apoptosis. Therefore, it is crucial to control inflammation and oxidation injury to minimize the nerve cell death during secondary spinal cord injury. Therefore, in response to oxidation and inflammation, heme oxygenase-1 (HO-1) is induced by the resident cells to ameliorate the milieu. In the meanwhile, neurotrophic factors are induced to promote neuroregeneration. However, it seems that anti-stress enzyme (HO-1) and neurotrophic factor (BDNF) do not significantly combat the pathological events during secondary spinal cord injury. Therefore, optimum healing can be induced if anti-inflammatory and neurotrophic factors are administered in a higher amount through an exogenous source. During the first experiment, the inflammation and neuroregeneration were selectively targeted. HO-1 expressing MSCs (HO-1 MSCs) and BDNF expressing MSCs (BDNF MSC) were co-transplanted in one group (combination group) of dogs with subacute spinal cord injury to selectively control the expression of inflammatory cytokines by HO-1 and induce neuroregeneration by BDNF. We compared the combination group with the HO-1 MSCs group, BDNF MSCs group, and GFP MSCs group. We found that the combination group showed significant improvement in functional recovery. It showed increased expression of neural markers and growth-associated proteins (GAP-43) than in other groups, which depicts enhanced neuroregeneration/neural sparing due to reduced expression of pro-inflammatory cytokines such as TNF-alpha, IL-6 and COX-2; and increased expression of anti-inflammatory markers such as IL-10 and HO-1. Histopathological study revealed reduced intra-parenchymal fibrosis in the injured spinal cord segment in the combination group than in other groups. Thus it was concluded that selectively targeting the inflammation and neuronal growth with the combined use of HO-1 MSCs and BDNF MSCs more favorably promote healing of the SCI. HO-1 MSCs play a role in controlling the inflammation, which favors the BDNF induced neuroregeneration at the injured spinal cord segment of dogs.

Keywords: HO-1 MSCs, BDNF MSCs, neuroregeneration, inflammation, anti-inflammation, spinal cord injury, dogs

Procedia PDF Downloads 119
40 Environmental Planning for Sustainable Utilization of Lake Chamo Biodiversity Resources: Geospatially Supported Approach, Ethiopia

Authors: Alemayehu Hailemicael Mezgebe, A. J. Solomon Raju

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Context: Lake Chamo is a significant lake in the Ethiopian Rift Valley, known for its diversity of wildlife and vegetation. However, the lake is facing various threats due to human activities and global effects. The poor management of resources could lead to food insecurity, ecological degradation, and loss of biodiversity. Research Aim: The aim of this study is to analyze the environmental implications of lake level changes using GIS and remote sensing. The research also aims to examine the floristic composition of the lakeside vegetation and propose spatially oriented environmental planning for the sustainable utilization of the biodiversity resources. Methodology: The study utilizes multi-temporal satellite images and aerial photographs to analyze the changes in the lake area over the past 45 years. Geospatial analysis techniques are employed to assess land use and land cover changes and change detection matrix. The composition and role of the lakeside vegetation in the ecological and hydrological functions are also examined. Findings: The analysis reveals that the lake has shrunk by 14.42% over the years, with significant modifications to its upstream segment. The study identifies various threats to the lake-wetland ecosystem, including changes in water chemistry, overfishing, and poor waste management. The study also highlights the impact of human activities on the lake's limnology, with an increase in conductivity, salinity, and alkalinity. Floristic composition analysis of the lake-wetland ecosystem showed definite pattern of the vegetation distribution. The vegetation composition can be generally categorized into three belts namely, the herbaceous belt, the legume belt and the bush-shrub-small trees belt. The vegetation belts collectively act as different-sized sieve screen system and calm down the pace of incoming foreign matter. This stratified vegetation provides vital information to decide the management interventions for the sustainability of lake-wetland ecosystem.Theoretical Importance: The study contributes to the understanding of the environmental changes and threats faced by Lake Chamo. It provides insights into the impact of human activities on the lake-wetland ecosystem and emphasizes the need for sustainable resource management. Data Collection and Analysis Procedures: The study utilizes aerial photographs, satellite imagery, and field observations to collect data. Geospatial analysis techniques are employed to process and analyze the data, including land use/land cover changes and change detection matrices. Floristic composition analysis is conducted to assess the vegetation patterns Question Addressed: The study addresses the question of how lake level changes and human activities impact the environmental health and biodiversity of Lake Chamo. It also explores the potential opportunities and threats related to water utilization and waste management. Conclusion: The study recommends the implementation of spatially oriented environmental planning to ensure the sustainable utilization and maintenance of Lake Chamo's biodiversity resources. It emphasizes the need for proper waste management, improved irrigation facilities, and a buffer zone with specific vegetation patterns to restore and protect the lake outskirt.

Keywords: buffer zone, geo-spatial, lake chamo, lake level changes, sustainable utilization

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39 Physical Aspects of Shape Memory and Reversibility in Shape Memory Alloys

Authors: Osman Adiguzel

Abstract:

Shape memory alloys take place in a class of smart materials by exhibiting a peculiar property called the shape memory effect. This property is characterized by the recoverability of two certain shapes of material at different temperatures. These materials are often called smart materials due to their functionality and their capacity of responding to changes in the environment. Shape memory materials are used as shape memory devices in many interdisciplinary fields such as medicine, bioengineering, metallurgy, building industry and many engineering fields. The shape memory effect is performed thermally by heating and cooling after first cooling and stressing treatments, and this behavior is called thermoelasticity. This effect is based on martensitic transformations characterized by changes in the crystal structure of the material. The shape memory effect is the result of successive thermally and stress-induced martensitic transformations. Shape memory alloys exhibit thermoelasticity and superelasticity by means of deformation in the low-temperature product phase and high-temperature parent phase region, respectively. Superelasticity is performed by stressing and releasing the material in the parent phase region. Loading and unloading paths are different in the stress-strain diagram, and the cycling loop reveals energy dissipation. The strain energy is stored after releasing, and these alloys are mainly used as deformation absorbent materials in control of civil structures subjected to seismic events, due to the absorbance of strain energy during any disaster or earthquake. Thermal-induced martensitic transformation occurs thermally on cooling, along with lattice twinning with cooperative movements of atoms by means of lattice invariant shears, and ordered parent phase structures turn into twinned martensite structures, and twinned structures turn into the detwinned structures by means of stress-induced martensitic transformation by stressing the material in the martensitic condition. Thermal induced transformation occurs with the cooperative movements of atoms in two opposite directions, <110 > -type directions on the {110} - type planes of austenite matrix which is the basal plane of martensite. Copper-based alloys exhibit this property in the metastable β-phase region, which has bcc-based structures at high-temperature parent phase field. Lattice invariant shear and twinning is not uniform in copper-based ternary alloys and gives rise to the formation of complex layered structures, depending on the stacking sequences on the close-packed planes of the ordered parent phase lattice. In the present contribution, x-ray diffraction and transmission electron microscopy (TEM) studies were carried out on two copper-based CuAlMn and CuZnAl alloys. X-ray diffraction profiles and electron diffraction patterns reveal that both alloys exhibit superlattice reflections inherited from the parent phase due to the displacive character of martensitic transformation. X-ray diffractograms taken in a long time interval show that diffraction angles and intensities of diffraction peaks change with the aging duration at room temperature. In particular, some of the successive peak pairs providing a special relation between Miller indices come close to each other. This result refers to the rearrangement of atoms in a diffusive manner.

Keywords: shape memory effect, martensitic transformation, reversibility, superelasticity, twinning, detwinning

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38 Stromal Vascular Fraction Regenerative Potential in a Muscle Ischemia/Reperfusion Injury Mouse Model

Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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37 Relevance of Dosing Time for Everolimus Toxicity in Respect to the Circadian P-Glycoprotein Expression in Mdr1a::Luc Mice

Authors: Narin Ozturk, Xiao-Mei Li, Sylvie Giachetti, Francis Levi, Alper Okyar

Abstract:

P-glycoprotein (P-gp, MDR1, ABCB1) is a transmembrane protein acting as an ATP-dependent efflux pump and functions as a biological barrier by extruding drugs and xenobiotics out of cells in healthy tissues especially in intestines, liver and brain as well as in tumor cells. The circadian timing system controls a variety of biological functions in mammals including xenobiotic metabolism and detoxification, proliferation and cell cycle events, and may affect pharmacokinetics, toxicity and efficacy of drugs. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer drug that is active against many cancers, and its pharmacokinetics depend on P-gp. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus with respect to the intestinal P-gp expression rhythms in mdr1a::Luc mice using Real Time-Biolumicorder (RT-BIO) System. Mdr1a::Luc male mice were synchronized with 12 h of Light and 12 h of Dark (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding Light onset). After 1-week baseline recordings, everolimus (5 mg/kg/day x 14 days) was administered orally at ZT1-resting period- and ZT13-activity period- to mdr1a::Luc mice singly housed in an innovative monitoring device, Real Time-Biolumicorder units which let us monitor real-time and long-term gene expression in freely moving mice. D-luciferin (1.5 mg/mL) was dissolved in drinking water. Mouse intestinal mdr1a::Luc oscillation profile reflecting P-gp gene expression and locomotor activity pattern were recorded every minute with the photomultiplier tube and infrared sensor respectively. General behavior and clinical signs were monitored, and body weight was measured every day as an index of toxicity. Drug-induced body weight change was expressed relative to body weight on the initial treatment day. Statistical significance of differences between groups was validated with ANOVA. Circadian rhythms were validated with Cosinor Analysis. Everolimus toxicity changed as a function of drug timing, which was least following dosing at ZT13, near the onset of the activity span in male mice. Mean body weight loss was nearly twice as large in mice treated with 5 mg/kg everolimus at ZT1 as compared to ZT13 (8.9% vs. 5.4%; ANOVA, p < 0.001). Based on the body weight loss and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13. Both rest-activity and mdr1a::Luc expression displayed stable 24-h periodic rhythms before everolimus and in both vehicle-treated controls. Real-time bioluminescence pattern of mdr1a revealed a circadian rhythm with a 24-h period with an acrophase at ZT16 (Cosinor, p < 0.001). Mdr1a expression remained rhythmic in everolimus-treated mice, whereas down-regulation was observed in P-gp expression in 2 of 4 mice. The study identified the circadian pattern of intestinal P-gp expression with an unprecedented precision. The circadian timing depending on the P-gp expression rhythms may play a crucial role in the tolerability/toxicity of everolimus. The circadian changes in mdr1a genes deserve further studies regarding their relevance for in vitro and in vivo chronotolerance of mdr1a-transported anticancer drugs. Chronotherapy with P-gp-effluxed anticancer drugs could then be applied according to their rhythmic patterns in host and tumor to jointly maximize treatment efficacy and minimize toxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, mdr1a::Luc mice, p-glycoprotein

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36 Thermal Characterisation of Multi-Coated Lightweight Brake Rotors for Passenger Cars

Authors: Ankit Khurana

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The sufficient heat storage capacity or ability to dissipate heat is the most decisive parameter to have an effective and efficient functioning of Friction-based Brake Disc systems. The primary aim of the research was to analyse the effect of multiple coatings on lightweight disk rotors surface which not only alleviates the mass of vehicle & also, augments heat transfer. This research is projected to aid the automobile fraternity with an enunciated view over the thermal aspects in a braking system. The results of the project indicate that with the advent of modern coating technologies a brake system’s thermal curtailments can be removed and together with forced convection, heat transfer processes can see a drastic improvement leading to increased lifetime of the brake rotor. Other advantages of modifying the surface of a lightweight rotor substrate will be to reduce the overall weight of the vehicle, decrease the risk of thermal brake failure (brake fade and fluid vaporization), longer component life, as well as lower noise and vibration characteristics. A mathematical model was constructed in MATLAB which encompassing the various thermal characteristics of the proposed coatings and substrate materials required to approximate the heat flux values in a free and forced convection environment; resembling to a real-time braking phenomenon which could easily be modelled into a full cum scaled version of the alloy brake rotor part in ABAQUS. The finite element of a brake rotor was modelled in a constrained environment such that the nodal temperature between the contact surfaces of the coatings and substrate (Wrought Aluminum alloy) resemble an amalgamated solid brake rotor element. The initial results obtained were for a Plasma Electrolytic Oxidized (PEO) substrate wherein the Aluminum alloy gets a hard ceramic oxide layer grown on its transitional phase. The rotor was modelled and then evaluated in real-time for a constant ‘g’ braking event (based upon the mathematical heat flux input and convective surroundings), which reflected the necessity to deposit a conducting coat (sacrificial) above the PEO layer in order to inhibit thermal degradation of the barrier coating prematurely. Taguchi study was then used to bring out certain critical factors which may influence the maximum operating temperature of a multi-coated brake disc by simulating brake tests: a) an Alpine descent lasting 50 seconds; b) an Autobahn stop lasting 3.53 seconds; c) a Six–high speed repeated stop in accordance to FMVSS 135 lasting 46.25 seconds. Thermal Barrier coating thickness and Vane heat transfer coefficient were the two most influential factors and owing to their design and manufacturing constraints a final optimized model was obtained which survived the 6-high speed stop test as per the FMVSS -135 specifications. The simulation data highlighted the merits for preferring Wrought Aluminum alloy 7068 over Grey Cast Iron and Aluminum Metal Matrix Composite in coherence with the multiple coating depositions.

Keywords: lightweight brakes, surface modification, simulated braking, PEO, aluminum

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35 Encapsulated Bioflavonoids: Nanotechnology Driven Food Waste Utilization

Authors: Niharika Kaushal, Minni Singh

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Citrus fruits fall into the category of those commercially grown fruits that constitute an excellent repository of phytochemicals with health-promoting properties. Fruits belonging to the citrus family, when processed by industries, produce tons of agriculture by-products in the form of peels, pulp, and seeds, which normally have no further usage and are commonly discarded. In spite of this, such residues are of paramount importance due to their richness in valuable compounds; therefore, agro-waste is considered a valuable bioresource for various purposes in the food sector. A range of biological properties, including anti-oxidative, anti-cancerous, anti-inflammatory, anti-allergenicity, and anti-aging activity, have been reported for these bioactive compounds. Taking advantage of these inexpensive residual sources requires special attention to extract bioactive compounds. Mandarin (Citrus nobilis X Citrus deliciosa) is a potential source of bioflavonoids with antioxidant properties, and it is increasingly regarded as a functional food. Despite these benefits, flavonoids suffer from a barrier of pre-systemic metabolism in gastric fluid, which impedes their effectiveness. Therefore, colloidal delivery systems can completely overcome the barrier in question. This study involved the extraction and identification of key flavonoids from mandarin biomass. Using a green chemistry approach, supercritical fluid extraction at 330 bar, temperature 40C, and co-solvent 10% ethanol was employed for extraction, and the identification of flavonoids was made by mass spectrometry. As flavonoids are concerned with a limitation, the obtained extract was encapsulated in polylactic-co-glycolic acid (PLGA) matrix using a solvent evaporation method. Additionally, the antioxidant potential was evaluated by the 2,2-diphenylpicrylhydrazyl (DPPH) assay. A release pattern of flavonoids was observed over time using simulated gastrointestinal fluids. From the results, it was observed that the total flavonoids extracted from the mandarin biomass were estimated to be 47.3 ±1.06 mg/ml rutin equivalents as total flavonoids. In the extract, significantly, polymethoxyflavones (PMFs), tangeretin and nobiletin were identified, followed by hesperetin and naringin. The designed flavonoid-PLGA nanoparticles exhibited a particle size between 200-250nm. In addition, the bioengineered nanoparticles had a high entrapment efficiency of nearly 80.0% and maintained stability for more than a year. Flavonoid nanoparticles showed excellent antioxidant activity with an IC50 of 0.55μg/ml. Morphological studies revealed the smooth and spherical shape of nanoparticles as visualized by Field emission scanning electron microscopy (FE-SEM). Simulated gastrointestinal studies of free extract and nanoencapsulation revealed the degradation of nearly half of the flavonoids under harsh acidic conditions in the case of free extract. After encapsulation, flavonoids exhibited sustained release properties, suggesting that polymeric encapsulates are efficient carriers of flavonoids. Thus, such technology-driven and biomass-derived products form the basis for their use in the development of functional foods with improved therapeutic potential and antioxidant properties. As a result, citrus processing waste can be considered a new resource that has high value and can be used for promoting its utilization.

Keywords: citrus, agrowaste, flavonoids, nanoparticles

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34 Organic Tuber Production Fosters Food Security and Soil Health: A Decade of Evidence from India

Authors: G. Suja, J. Sreekumar, A. N. Jyothi, V. S. Santhosh Mithra

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Worldwide concerns regarding food safety, environmental degradation and threats to human health have generated interest in alternative systems like organic farming. Tropical tuber crops, cassava, sweet potato, yams, and aroids are food-cum-nutritional security-cum climate resilient crops. These form stable or subsidiary food for about 500 million global population. Cassava, yams (white yam, greater yam, and lesser yam) and edible aroids (elephant foot yam, taro, and tannia) are high energy tuberous vegetables with good taste and nutritive value. Seven on-station field experiments at ICAR-Central Tuber Crops Research Institute, Thiruvananthapuram, India and seventeen on-farm trials in three districts of Kerala, were conducted over a decade (2004-2015) to compare the varietal response, yield, quality and soil properties under organic vs conventional system in these crops and to develop a learning system based on the data generated. The industrial, as well as domestic varieties of cassava, the elite and local varieties of elephant foot yam and taro and the three species of Dioscorea (yams), were on a par under both systems. Organic management promoted yield by 8%, 20%, 9%, 11% and 7% over conventional practice in cassava, elephant foot yam, white yam, greater yam and lesser yam respectively. Elephant foot yam was the most responsive to organic management followed by yams and cassava. In taro, slight yield reduction (5%) was noticed under organic farming with almost similar tuber quality. The tuber quality was improved with higher dry matter, starch, crude protein, K, Ca and Mg contents. The anti-nutritional factors, oxalate content in elephant foot yam and cyanogenic glucoside content in cassava were lowered by 21 and 12.4% respectively. Organic plots had significantly higher water holding capacity, pH, available K, Fe, Mn and Cu, higher soil organic matter, available N, P, exchangeable Ca and Mg, dehydrogenase enzyme activity and microbial count. Organic farming scored significantly higher soil quality index (1.93) than conventional practice (1.46). The soil quality index was driven by water holding capacity, pH and available Zn followed by soil organic matter. Organic management enhanced net profit by 20-40% over chemical farming. A case in point is the cost-benefit analysis in elephant foot yam which indicated that the net profit was 28% higher and additional income of Rs. 47,716 ha-1 was obtained due to organic farming. Cost-effective technologies were field validated. The on-station technologies developed were validated and popularized through on-farm trials in 10 sites (5 ha) under National Horticulture Mission funded programme in elephant foot yam and seven sites in yams and taro. The technologies are included in the Package of Practices Recommendations for crops of Kerala Agricultural University. A learning system developed using artificial neural networks (ANN) predicted the performance of elephant foot yam organic system. Use of organically produced seed materials, seed treatment in cow-dung, neem cake, bio-inoculant slurry, farmyard manure incubated with bio-inoculants, green manuring, use of neem cake, bio-fertilizers and ash formed the strategies for organic production. Organic farming is an eco-friendly management strategy that enables 10-20% higher yield, quality tubers and maintenance of soil health in tuber crops.

Keywords: eco-agriculture, quality, root crops, healthy soil, yield

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33 Multibody Constrained Dynamics of Y-Method Installation System for a Large Scale Subsea Equipment

Authors: Naeem Ullah, Menglan Duan, Mac Darlington Uche Onuoha

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The lowering of subsea equipment into the deep waters is a challenging job due to the harsh offshore environment. Many researchers have introduced various installation systems to deploy the payload safely into the deep oceans. In general practice, dual floating vessels are not employed owing to the prevalent safety risks and hazards caused by ever-increasing dynamical effects sourced by mutual interaction between the bodies. However, while keeping in the view of the optimal grounds, such as economical one, the Y-method, the two conventional tugboats supporting the equipment by the two independent strands connected to a tri-plate above the equipment, has been employed to study multibody dynamics of the dual barge lifting operations. In this study, the two tugboats and the suspended payload (Y-method) are deployed for the lowering of subsea equipment into the deep waters as a multibody dynamic system. The two-wire ropes are used for the lifting and installation operation by this Y-method installation system. 6-dof (degree of freedom) for each body are considered to establish coupled 18-dof multibody model by embedding technique or velocity transformation technique. The fundamental and prompt advantage of this technique is that the constraint forces can be eliminated directly, and no extra computational effort is required for the elimination of the constraint forces. The inertial frame of reference is taken at the surface of the water as the time-independent frame of reference, and the floating frames of reference are introduced in each body as the time-dependent frames of reference in order to formulate the velocity transformation matrix. The local transformation of the generalized coordinates to the inertial frame of reference is executed by applying the Euler Angle approach. The spherical joints are articulated amongst the multibody as the kinematic joints. The hydrodynamic force, the two-strand forces, the hydrostatic force, and the mooring forces are taken into consideration as the external forces. The radiation force of the hydrodynamic force is obtained by employing the Cummins equation. The wave exciting part of the hydrodynamic force is obtained by using force response amplitude operators (RAOs) that are obtained by the commercial solver ‘OpenFOAM’. The strand force is obtained by considering the wire rope as an elastic spring. The nonlinear hydrostatic force is obtained by the pressure integration technique at each time step of the wave movement. The mooring forces are evaluated by using Faltinsen analytical approach. ‘The Runge Kutta Method’ of Fourth-Order is employed to evaluate the coupled equations of motion obtained for 18-dof multibody model. The results are correlated with the simulated Orcaflex Model. Moreover, the results from Orcaflex Model are compared with the MOSES Model from previous studies. The MBDS of single barge lifting operation from the former studies are compared with the MBDS of the established dual barge lifting operation. The dynamics of the dual barge lifting operation are found larger in magnitude as compared to the single barge lifting operation. It is noticed that the traction at the top connection point of the cable decreases with the increase in the length, and it becomes almost constant after passing through the splash zone.

Keywords: dual barge lifting operation, Y-method, multibody dynamics, shipbuilding, installation of subsea equipment, shipbuilding

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32 Effect of Selenium Source on Meat Quality of Bonsmara Bull Calves

Authors: J. van Soest, B. Bruneel, J. Smit, N. Williams, P. Swiegers

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Selenium (Se) is an essential trace mineral involved in reducing oxidative stress, enhancing immune status, improving reproduction, and regulating growth. During finishing period, selenium supplementation can be applied to improve meat quality. Dietary selenium can be provided in inorganic or organic forms. Specifically, L-selenomethionine (organic selenium) allows for selenium storage in animal protein which supports the animal during periods of high oxidative stress. The objective of this study was to investigate the effects of synthetically produced, single amino acid, L-selenomethionine (Excential Selenium 4000, Orffa Additives BV) on production parameters, health status, and meat quality of Bonsmara bull calves. 24 calves, 7 months of age, completed a 60-day initial growing period at a commercial feedlot, after which they were transported to research station Rumen-8 (Bethlehem, South-Africa). After a ten-day adaptation period, the bulls were allocated to a control (n=12) or treatment (n=12) group. Each group was divided over 3 pens based on weight. Both groups received Total Mixed Ration supplemented with 5.25 mg Se/head per day. The control group was supplemented with sodium selenite as Se source, whilst the treatment group was supplemented with L-selenomethionine (Excential Selenium 4000, Orffa Additives BV). Animals were limited to 10 kg feed intake per head per day to ensure similar Se intake. Treatment period lasted 1.5 months. A beta-adrenergic agonist was included in the feed for the last 30 days. During the treatment period, average daily gain, average daily feed intake, and feed conversion ratio were recorded. Blood parameters were measured at day 1, day 25, and before slaughter (day 47). After slaughter, carcass weight, dressing percentage, grading, and meat quality (pH, tenderness, colour, odour, purge, proximate analyses, acid detergent fibre, and neutral detergent fibre) were determined. No differences between groups were found in performance. A higher number of animals with cortisol levels below detection limit (27.6 nmol/l) was recorded for the treatment group. Other blood parameters showed no differences. No differences were found regarding carcass weight and dressing percentage. Important parameters of meat quality were significantly improved in the treatment group: instrumental tenderness at 14 days ageing was 2.8 and 3.4 for treatment and control respectively (P=0.010), and a 0.5% decrease in purge (of fresh samples) was shown, 1.5% and 2.0% for treatment group and control respectively (p=0.029). Besides, pH was shown to be numerically reduced in the treatment group. In summary, supplementation with L-selenomethionine as selenium source improved meat quality compared to sodium selenite. Lower instrumental tenderness (Warner Bratzler Shear Force, WBSF) was recorded for the treatment group. This indicates less tough meat and highest consumer satisfaction. Regarding purge, control was just below 2.0%, an important threshold for consumer acceptation. Treatment group scored 0.5% lower for purge than control, indicating higher consumer satisfaction. The lower pH in the treatment group could be an indication of higher glycogen reserves in muscle which could contribute to a reduced risk of Dark Firm Dry carcasses. More animals showed cortisol levels below detection limit in the treatment group, indicating lower levels of stress when animals receive L-selenomethionine.

Keywords: calves, meat quality, nutrition, selenium

Procedia PDF Downloads 183
31 A Descriptive Study on Water Scarcity as a One Health Challenge among the Osiram Community, Kajiado County, Kenya

Authors: Damiano Omari, Topirian Kerempe, Dibo Sama, Walter Wafula, Sharon Chepkoech, Chrispine Juma, Gilbert Kirui, Simon Mburu, Susan Keino

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The One Health concept was officially adopted by the international organizations and scholarly bodies in 1984. It aims at combining human, animal and environmental components to address global health challenges. Using collaborative efforts optimal health to people, animals, and the environment can be achieved. One health approach plays a significant approach role in prevention and control of zoonosis diseases. It has also been noted that 75% of new emerging human infectious diseases are zoonotic. In Kenya, one health has been embraced and strongly advocated for by One Health East and Central Africa (OHCEA). It was inaugurated on 17th of October 2010 at a historic meeting facilitated by USAID with participants from 7 public health schools, seven faculties of veterinary medicine in Eastern Africa and 2 American universities (Tufts and University of Minnesota) in addition to respond project staff. The study was conducted in Loitoktok Sub County, specifically in the Amboseli Ecosystem. The Amboseli ecosystem covers an area of 5,700 square kilometers and stretches between Mt. Kilimanjaro, Chyulu Hills, Tsavo West National park and the Kenya/Tanzania border. The area is arid to semi-arid and is more suitable for pastoralism with a high potential for conservation of wildlife and tourism enterprises. The ecosystem consists of the Amboseli National Park, which is surrounded by six group ranches which include Kimana, Olgulului, Selengei, Mbirikani, Kuku and Rombo in Loitoktok District. The Manyatta of study was Osiram Cultural Manyatta in Mbirikani group ranch. Apart from visiting the Manyatta, we also visited the sub-county hospital, slaughter slab, forest service, Kimana market, and the Amboseli National Park. The aim of the study was to identify the one health issues facing the community. This was done by a conducting a community needs assessment and prioritization. Different methods were used in data collection for the qualitative and numerical data. They include among others; key informant interviews and focus group discussions. We also guided the community members in drawing their Resource Map this helped identify the major resources in their land and also help them identify some of the issues they were facing. Matrix piling, root cause analysis, and force field analysis tools were used to establish the one health related priority issues facing community members. Skits were also used to present to the community interventions to the major one health issues. Some of the prioritized needs among the community were water scarcity and inadequate markets for their beadwork. The group intervened on the various needs of the Manyatta. For water scarcity, we educated the community on water harvesting methods using gutters as well as proper storage by the use of tanks and earth dams. The community was also encouraged to recycle and conserve water. To improve markets; we educated the community to upload their products online, a page was opened for them and uploading the photos was demonstrated to them. They were also encouraged to be innovative to attract more clients.

Keywords: Amboseli ecosystem, community interventions, community needs assessment and prioritization, one health issues

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30 An Intelligence-Led Methodologly for Detecting Dark Actors in Human Trafficking Networks

Authors: Andrew D. Henshaw, James M. Austin

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Introduction: Human trafficking is an increasingly serious transnational criminal enterprise and social security issue. Despite ongoing efforts to mitigate the phenomenon and a significant expansion of security scrutiny over past decades, it is not receding. This is true for many nations in Southeast Asia, widely recognized as the global hub for trafficked persons, including men, women, and children. Clearly, human trafficking is difficult to address because there are numerous drivers, causes, and motivators for it to persist, such as non-military and non-traditional security challenges, i.e., climate change, global warming displacement, and natural disasters. These make displaced persons and refugees particularly vulnerable. The issue is so large conservative estimates put a dollar value at around $150 billion-plus per year (Niethammer, 2020) spanning sexual slavery and exploitation, forced labor, construction, mining and in conflict roles, and forced marriages of girls and women. Coupled with corruption throughout military, police, and civil authorities around the world, and the active hands of powerful transnational criminal organizations, it is likely that such figures are grossly underestimated as human trafficking is misreported, under-detected, and deliberately obfuscated to protect those profiting from it. For example, the 2022 UN report on human trafficking shows a 56% reduction in convictions in that year alone (UNODC, 2022). Our Approach: To better understand this, our research utilizes a bespoke methodology. Applying a JAM (Juxtaposition Assessment Matrix), which we previously developed to detect flows of dark money around the globe (Henshaw, A & Austin, J, 2021), we now focus on the human trafficking paradigm. Indeed, utilizing a JAM methodology has identified key indicators of human trafficking not previously explored in depth. Being a set of structured analytical techniques that provide panoramic interpretations of the subject matter, this iteration of the JAM further incorporates behavioral and driver indicators, including the employment of Open-Source Artificial Intelligence (OS-AI) across multiple collection points. The extracted behavioral data was then applied to identify non-traditional indicators as they contribute to human trafficking. Furthermore, as the JAM OS-AI analyses data from the inverted position, i.e., the viewpoint of the traffickers, it examines the behavioral and physical traits required to succeed. This transposed examination of the requirements of success delivers potential leverage points for exploitation in the fight against human trafficking in a new and novel way. Findings: Our approach identified new innovative datasets that have previously been overlooked or, at best, undervalued. For example, the JAM OS-AI approach identified critical 'dark agent' lynchpins within human trafficking that are difficult to detect and harder to connect to actors and agents within a network. Our preliminary data suggests this is in part due to the fact that ‘dark agents’ in extant research have been difficult to detect and potentially much harder to directly connect to the actors and organizations in human trafficking networks. Our research demonstrates that using new investigative techniques such as OS-AI-aided JAM introduces a powerful toolset to increase understanding of human trafficking and transnational crime and illuminate networks that, to date, avoid global law enforcement scrutiny.

Keywords: human trafficking, open-source intelligence, transnational crime, human security, international human rights, intelligence analysis, JAM OS-AI, Dark Money

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29 Effects of Delphinidin on Lipid Metabolism in HepG2 Cells and Diet-Induced Obese Mice

Authors: Marcela Parra-Vargas, Ana Sandoval-Rodriguez, Roberto Rodriguez-Echevarria, Jose Dominguez-Rosales, Juan Armendariz-Borunda

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Non-alcoholic fatty liver disease (NAFLD) is characterized by an excess of hepatic lipids, and it is to author’s best knowledge, the most prevalent chronic liver disorder. Anthocyanin-rich food consumption is linked to health benefits in metabolic disorders associated with obesity and NAFLD, although the precise functional role of anthocyanidin delphinidin (Dp) has yet to be established. The aim of this study was to investigate the effect of the Dp in NAFLD metabolic alterations by evaluating prevention or amelioration of hepatic lipid accumulation, as well as molecular mechanisms in two experimental obesity-related models of NALFD. In vitro: HepG2 cells were incubated with sodium palmitate (PA, 1 mM) to induce lipotoxic damage, and concomitantly treated with Dp (180 uM) for 24 h. Subsequently, total lipid accumulation was measured by colorimetric staining with Oil Red O, and total intrahepatic triglycerides were determined by an enzymatic assay. To assess molecular mechanisms, cells were pre-treated with PA for 24 h and then exposed to Dp for 1 h. In vivo: four-week-old male C57BL/6Nhsd mice were allocated in two main groups. Mice were fed with standard diet (control) or high-fat and high-carbohydrate diet (45% fat, HFD) for 16 wk to induce NAFLD. Then HFD was divided into subgroups: one treated orally with Dp (15 mg/kg bw, HFD-Dp) every day for 4 wk, while HFD group treated with vehicle (DMSO). Weight and fasting glucose were recorded weekly, while dietary ingestion was measured daily. Insulin tolerance test was performed at the end of treatment. Liver histology was evaluated with H&E and Masson’s trichrome stain. RT-PCR was used to evaluate gene expression and Western Blot to determine levels of protein in both experimental models. Parametric data were analyzed with one-way ANOVA and Tukey’s post-hoc test. Kruskal-Wallis and Mann-Whitney U test for non-parametric data, and P < 0.5 were considered significant. Dp prevented hepatic lipid accumulation by PA in HepG2 hepatocytes. Furthermore, Dp down-regulated gene expression of SREBP1c, FAS, and CPT1a without modifying AMPK phosphorylation levels. In vivo, Dp oral administration did not ameliorate lipid metabolic alterations raised by HFD. Adiposity, dietary ingestion, fasting glucose, and insulin sensitivity after Dp treatment remained similar to HFD group. Histological analysis showed hepatic damage in HFD groups and no differences between HFD and HFD-Dp groups were found. Hepatic gene expression of ACC and FAS were not altered by HFD. SREBP1c was similar in both HFD and HFD-Dp groups. No significant changes were observed in SREBP1c, ACC, and FAS adipose tissue gene expression by HFD or Dp treatment. Additionally, immunoblotting analysis revealed no changes in pathway SIRT1-LKB-AMPK and PPAR alpha by both HFD groups compared to control. In conclusion, the antioxidant Dp may provoke beneficial effects in the prevention of hepatic lipid accumulation. Nevertheless, the oral dose administrated in mice that simulated the total intake of anthocyanins consumed daily by humans has no effect as a treatment on hepatic lipid metabolic alterations and histological abnormalities associated with exposure to chronic HFD. A healthy lifestyle with regular intake of antioxidants such as anthocyanins may prevent metabolic alterations in NAFLD.

Keywords: anthocyanins, antioxidants, delphinidin, non-alcoholic fatty liver disease, obesity

Procedia PDF Downloads 202