Search results for: direct immunofluorescence assay (DFA)

Authors: O. Gsib, U. Peirera, C. Egles, S. A. Bencherif

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In skin regenerative medicine, one of the critical issues is to produce a three-dimensional scaffold with optimized porosity for dermal fibroblast infiltration and neovascularization, which exhibits high mechanical properties and displays sufficient wound healing characteristics. In this study, we report on the synthesis and characterization of macroporous sequential interpenetrating polymer networks (IPNs) combining skin wound healing properties of fibrin with the excellent physical properties of polyethylene glycol (PEG). Fibrin fibers serve as a provisional biologically active network to promote cell adhesion and proliferation while PEG provides the mechanical stability to maintain the entire 3D construct. After having modified both PEG and Serum Albumin (used for promoting enzymatic degradability) by adding methacrylate residues (PEGDM and SAM, respectively), Fibrin/PEGDM-SAM sequential IPNs were synthesized as follows: Macroporous sponges were first produced from PEGDM-SAM hydrogels by a freeze-drying technique and then rehydrated by adding the fibrin precursors. Environmental Scanning Electron Microscopy (ESEM) and Confocal Laser Scanning Microscopy (CLSM) were used to characterize their microstructure. Human dermal fibroblasts were cultivated during one week in the constructs and different cell culture parameters (viability, morphology, proliferation) were evaluated. Subcutaneous implantations of the scaffolds were conducted on five-week old male nude mice to investigate their biocompatibility in vivo. We successfully synthesized interconnected and macroporous Fibrin/PEGDM-SAM sequential IPNs. The viability of primary dermal fibroblasts was well maintained (above 90%) after 2 days of culture. Cells were able to adhere, spread and proliferate in the scaffolds suggesting the suitable porosity and intrinsic biologic properties of the constructs. The fibrin network adopted a spider web shape that covered partially the pores allowing easier cell infiltration into the macroporous structure. To further characterize the in vitro cell behavior, cell proliferation (EdU incorporation, MTS assay) is being studied. Preliminary histological analysis of animal studies indicated the persistence of hydrogels even after one-month post implantation and confirmed the absence of inflammation response, good biocompatibility and biointegration of our scaffolds within the surrounding tissues. These results suggest that our Fibrin/PEGDM-SAM IPNs could be considered as potential candidates for dermis regenerative medicine. Histological analysis will be completed to further assess scaffold remodeling including de novo extracellular matrix protein synthesis and early stage angiogenesis analysis. Compression measurements will be conducted to investigate the mechanical properties.

Keywords: fibrin, hydrogels for dermal reconstruction, polyethylene glycol, semi-interpenetrating polymer network

Procedia PDF Downloads 230

Authors: Jin Hwan Cheong, Mina Hwang, Myung Hoon Han, Je Il Ryu, Young ha Oh, Seong Ho Koh, Wu Duck Won, Byung Jin Ha

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Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) has been reported to play critical roles in the proliferation of various cancer cells. However, the roles of LGR5 in brain tumors and the specific intracellular signaling proteins directly associated with it remain unknown. Expression of LGR5 was first measured in normal brain tissue, meningioma, and pituitary adenoma of humans. To identify the downstream signaling pathways of LGR5, siRNA-mediated knockdown of LGR5 was performed in SH-SY5Y neuroblastoma cells followed by proteomics analysis with 2-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In addition, the expression of LGR5-associated proteins was evaluated in LGR5-inꠓhibited neuroblastoma cells and in human normal brain, meningioma, and pituitary adenoma tissue. Proteomics analysis showed 12 protein spots were significantly different in expression level (more than two-fold change) and subsequently identified by peptide mass fingerprinting. A protein association network was constructed from the 12 identified proteins altered by LGR5 knockdown. Direct and indirect interactions were identified among the 12 proteins. HSP 90-beta was one of the proteins whose expression was altered by LGR5 knockdown. Likewise, we observed decreased expression of proteins in the hnRNP subfamily following LGR5 knockdown. In addition, we have for the first time identified significantly higher hnRNP family expression in meningioma and pituitary adenoma compared to normal brain tissue. Taken together, LGR5 and its downstream sigꠓnaling play critical roles in neuroblastoma and brain tumors such as meningioma and pituitary adenoma.

Keywords: LGR5, neuroblastoma, meningioma, pituitary adenoma, hnRNP

Procedia PDF Downloads 47

Authors: Mohammad Zafar-ul Islam, Shaily Menon, Xingong Li, A. Townsend Peterson

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In addition to the mounting empirical data on direct implications of climate change for natural and human systems, evidence is increasing for other, indirect climate change phenomena such as sea-level rise. Rising sea levels and associated marine intrusion into terrestrial environments are predicted to be among the most serious eventual consequences of climate change. The many complex and interacting factors affecting sea levels create considerable uncertainty in sea-level rise projections: conservative estimates are on the order of 0.5-1.0 m globally, while other estimates are much higher, approaching 6 m. Marine intrusion associated with 1– 6 m sea-level rise will impact species and habitats in coastal ecosystems severely. Examining areas most vulnerable to such impacts may allow design of appropriate adaptation and mitigation strategies. We present an overview of potential effects of 1 and 6 m sea level rise for coastal conservation areas in the Indian Subcontinent. In particular, we examine the projected magnitude of areal losses in relevant biogeographic zones, ecoregions, protected areas (PAs), and Important Bird Areas (IBAs). In addition, we provide a more detailed and quantitative analysis of likely effects of marine intrusion on 22 coastal PAs and IBAs that provide critical habitat for birds in the form of breeding areas, migratory stopover sites, and overwintering habitats. Several coastal PAs and IBAs are predicted to experience higher than 50% losses to marine intrusion. We explore consequences of such inundation levels on species and habitat in these areas.

Keywords: sea-level change, coastal inundation, marine intrusion, biogeographic zones, ecoregions, protected areas, important bird areas, adaptation, mitigation

Procedia PDF Downloads 251

Authors: R. Morbidelli, C. Saltalippi, A. Flammini, A. Garcia-Marin, J. L. Ayuso-Munoz

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The climate change, mainly due to greenhouse gas emissions associated to human activities, has been modifying hydrologic processes with a direct effect on air surface temperature that has significantly increased in the last century at global scale. In this context the Mediterranean area is considered to be particularly sensitive to the climate change impacts on temperature indices. An analysis finalized to study the evolution of temperature indices and to check the existence of significant trends in the Umbria Region (Italy) is presented. Temperature data were obtained by seven meteorological stations uniformly distributed in the study area and characterized by very long series of temperature observations (at least 60 years) spanning the 1924-2015 period. A set of 39 temperature indices represented by monthly and annual mean, average maximum and average minimum temperatures, has been derived. The trend analysis was realized by applying the non-parametric Mann-Kendall test, while the non-parametric Pettit test and the parametric Standard Normal Homogeneity test (SNHT) were used to check the presence of breakpoints or in-homogeneities due to environmental changes/anthropic activity or climate change effects. The Umbria region, in agreement with other recent studies exploring the temperature behavior in Italy, shows a general increase in all temperature indices, with the only exception of Gubbio site that exhibits very light negative trends or absence of trend. The presence of break points and in-homogeneity was widely explored through the selected tests and the results were checked on the basis of the well-known metadata of the meteorological stations.

Keywords: reception theory, reading, literary translation, horizons of expectation, reader

Procedia PDF Downloads 154

Authors: Gi-Hoon Yang, JongHan Ha, MyungGu Yeo, JaeYoon Lee, SeungHyun Ahn, Hyeongjin Lee, HoJun Jeon, YongBok Kim, Minseong Kim, GeunHyung Kim

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Tissue engineering scaffolds must be biocompatible and biodegradable, provide adequate mechanical strength and cell attachment site for proliferation and differentiation. Furthermore, the scaffold morphology (such as pore size, porosity and pore interconnectivity) plays an important role. The electrospinning process has been widely used to fabricate micro/nano-sized fibres. Electrospinning allows for the fabrication of non-woven meshes containing micro- to nano-sized fibers providing high surface-to-volume area for cell attachment. Due to its advantageous characteristics, electrospinning is a useful method for skin, cartilage, bone, and nerve regeneration. In this study, we fabricated PCL scaffolds (SP) consisting of spiral-like struts using 3D melt-plotting system and micro/nanofibers using direct electrospinning writing. By altering the conditions of the conventional melt-plotting method, spiral-like struts were generated. Then, micro/nanofibers were deposited selectively. The control scaffold composed of perpendicular PCL struts was fabricated using the conventional melt-plotting method to compare the cellular activities. The effect on the attached cells (osteoblast-like cells (MG63)) was evaluated depending on the bending instability of the struts. The SP scaffolds showed enhanced biological properties such as initial cell attachment, proliferation and osteogenic differentiation. These results suggest that the SP scaffolds has potential as a bioengineered substitute for soft and hard tissue regeneration.

Keywords: cell attachment, electrospinning, mechanical strength, melt-plotting

Procedia PDF Downloads 313

Authors: Yuvraj Singh Dangi, Brajesh Kumar Tiwari, Ashok Kumar Jain, Kamta Prasad Namdeo

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The potential use of liposomes as drug carriers by i.v. injection is limited by their low stability in blood stream. Firstly, phospholipid exchange and transfer to lipoproteins, mainly HDL destabilizes and disintegrates liposomes with subsequent loss of content. To avoid the pain associated with injection and to obtain better patient compliance studies concerning various dosage forms, have been developed. Conventional liposomes (unilamellar and multilamellar) have certain drawbacks like low entrapment efficiency, stability and release of drug after single breach in external membrane, have led to the new type of liposomal systems. The challenge has been successfully met in the form of Double Liposomes (DL). DL is a recently developed type of liposome, consisting of smaller liposomes enveloped in lipid bilayers. The outer lipid layer of DL can protect inner liposomes against various enzymes, therefore DL was thought to be more effective than ordinary liposomes. This concept was also supported by in vitro release characteristics i.e. DL formation inhibited the release of drugs encapsulated in inner liposomes. DL consists of several small liposomes encapsulated in large liposomes, i.e., multivesicular vesicles (MVV), therefore, DL should be discriminated from ordinary classification of multilamellar vesicles (MLV), large unilamellar vesicles (LUV), small unilamellar vesicles (SUV). However, for these liposomes, the volume of inner phase is small and loading volume of water-soluble drugs is low. In the present study, the potential of phosphatidylethanolamine (PE) lipid anchored double liposomes (DL) to incorporate two drugs in a single system is exploited as a tool to augment the H. pylori eradication rate. Preparation of DL involves two steps, first formation of primary (inner) liposomes by thin film hydration method containing one drug, then addition of suspension of inner liposomes on thin film of lipid containing the other drug. The success of formation of DL was characterized by optical and transmission electron microscopy. Quantitation of DL-bacterial interaction was evaluated in terms of percent growth inhibition (%GI) on reference strain of H. pylori ATCC 26695. To confirm specific binding efficacy of DL to H. pylori PE surface receptor we performed an agglutination assay. Agglutination in DL treated H. pylori suspension suggested selectivity of DL towards the PE surface receptor of H. pylori. Monotherapy is generally not recommended for treatment of a H. pylori infection due to the danger of development of resistance and unacceptably low eradication rates. Therefore, combination therapy with amoxicillin trihydrate (AMOX) as anti-H. pylori agent and ranitidine bismuth citrate (RBC) as antisecretory agent were selected for the study with an expectation that this dual-drug delivery approach will exert acceptable anti-H. pylori activity.

Keywords: Helicobacter pylorI, amoxicillin trihydrate, Ranitidine Bismuth citrate, phosphatidylethanolamine, multi vesicular systems

Procedia PDF Downloads 202

Authors: Fergus Wade, Jasmine Makker, Matthew Jankinson, Aminah Qamar, Gemma Morrelli, Shayan Shah

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Background: Ambulance offload delays (AODs) affect patient outcomes. At baseline, the average AOD at Princess Royal University Hospital (PRUH) was 41 minutes, in breach of the 15-minute target. Aims: By February 2023, we aimed to reduce: the average AOD to 30 minutes percentage of AOD >30 minutes (PA30) to 25% and >60 minutes (PA60) to 10% Methods: Following a root-cause analysis, we implemented 2 Plan, Do, Study, Act (PDSA) cycles. PDSA-1 ‘Drop-and-run’: ambulances waiting >15 minutes for a handover left the patients in the Emergency Department (ED) and returned to the community. PDSA-2: Booking in the patients before the handover, allowing direct updates to online records, eliminating the need for handwritten notes. Outcome measures: AOD, PA30, and PA60, and process measures: total ambulances and patients in the ED were recorded for 16 weeks. Results: In PDSA-1, all parameters increased slightly despite unvarying ED crowding. In PDSA-2, two shifts in data were seen: initially, a sharp increase in the outcome measures consistent with increased ED crowding, followed by a downward shift when crowding returned to baseline (p<0.01). Within this interval, the AOD reduced to 29.9 minutes, and PA30 and PA60 were 31.2% and 9.2% respectively. Discussion/conclusion: PDSA-1 didn’t result in any significant changes; lack of compliance was a key cause. The initial upward shift in PDSA-2 is likely associated with NHS staff strikes. However, during the second interval, the AOD and the PA60 met our targets of 30 minutes and 10%, respectively, improving patient flow in the ED. This was sustained without further input and if maintained, saves 2 paramedic shifts every 3 days.

Keywords: ambulance offload, district general hospital, handover, quality improvement

Procedia PDF Downloads 94

Authors: J. Prathap Kumar, G. Vaitheeswaran

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The present study investigates the role of ammonium and potassium ion in the electronic, bonding and optical properties of dinitramide salts due to their stability and non-toxic nature. A detailed analysis of bonding between NH₄ and K with dinitramide, optical transitions from the valence band to the conduction band, absorption spectra, refractive indices, reflectivity, loss function are reported. These materials are well known as oxidizers in solid rocket propellants. In the present work, we use full potential linear augmented plane wave (FP-LAPW) method which is implemented in the Wien2k package within the framework of density functional theory. The standard DFT functional local density approximation (LDA) and generalized gradient approximation (GGA) always underestimate the band gap by 30-40% due to the lack of derivative discontinuities of the exchange-correlation potential with respect to an occupation number. In order to get reliable results, one must use hybrid functional (HSE-PBE), GW calculations and Tran-Blaha modified Becke-Johnson (TB-mBJ) potential. It is very well known that hybrid functionals GW calculations are very expensive, the later methods are computationally cheap. The new developed TB-mBJ functionals use information kinetic energy density along with the charge density employed in DFT. The TB-mBJ functionals cannot be used for total energy calculations but instead yield very much improved band gap. The obtained electronic band gap at gamma point for both the ammonium dinitramide and potassium dinitramide are found to be 2.78 eV and 3.014 eV with GGA functional, respectively. After the inclusion of TB-mBJ, the band gap improved by 4.162 eV for potassium dinitramide and 4.378 eV for ammonium dinitramide. The nature of the band gap is direct in ADN and indirect in KDN. The optical constants such as dielectric constant, absorption, and refractive indices, birefringence values are presented. Overall as there are no experimental studies we present the improved band gap with TB-mBJ functional following with optical properties.

Keywords: ammonium dinitramide, potassium dinitramide, DFT, propellants

Procedia PDF Downloads 149

Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

Procedia PDF Downloads 104

Authors: Nik Kamariah Nikmat, Noor Hasmini Abdghani

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The Malaysian government had consistently revived its campaign for “Buy Malaysian Goods” from time to time. The purpose of the campaign is to remind consumers to be ethnocentric and patriotic when purchasing product and services. This is necessary to ensure high demand for local products and services compared to foreign products. However, the decline of domestic investment in 2012 has triggered concern for the Malaysian economy. Hence, this study attempts to determine the drivers of actual purchasing behavior, intention to purchase domestic products and ethnocentrism. The study employs the cross-sectional primary data, self-administered on household, selected using stratified random sampling in four Malaysian regions. A nine factor driver of actual domestic purchasing behavior (culture openness, conservatism, collectivism, patriotism, control belief, interest in foreign travel, attitude, ethnocentrism and intention) were measured utilizing 60 items, using 7-point Likert-scale. From 1000 questionnaires distributed, a sample of 486 were returned representing 48.6 percent response rate. From the fit generated structural model (SEM analysis), it was found that the drivers of actual purchase behavior are collectivism, cultural openness and patriotism; the drivers of intention to purchase domestic product are attitude, control belief, collectivism and conservativeness; and drivers of ethnocentrism are cultural openness, control belief, foreign travel and patriotism. It also shows that Malaysian consumers scored high in ethnocentrism and patriotism. The findings are discussed in the perspective of its implication to Malaysian National Agenda.

Keywords: actual purchase, ethnocentrism, patriotism, culture openness, conservatism

Procedia PDF Downloads 310

Authors: Sakshi V. Tantak, Vishap K. Malik, Neelanjney Pilarisetty

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A patent is an exclusive right granted for an invention. It can be a product or a process that provides an innovative method of doing something, or offers a new technical perspective or solution to a problem. A patent can be obtained by making the technical information and details about the invention publicly available. The patent owner has exclusive rights to prevent or stop anyone from using the patented invention for commercial uses. Any commercial usage, distribution, import or export of a patented invention or product requires the patent owner’s consent. It has been observed that the central and important parts of patents are scripted in idiosyncratic and complex linguistic structures that can be difficult to read, comprehend or interpret for the masses. The abstracts of these patents tend to obfuscate the precise nature of the patent instead of clarifying it via direct and simple linguistic constructs. This makes it necessary to have an efficient access to this knowledge via concise and transparent summaries. However, as mentioned above, due to complex and repetitive linguistic constructs and extremely long sentences, common extraction-oriented automatic text summarization methods should not be expected to show a remarkable performance when applied to patent documents. Other, more content-oriented or abstractive summarization techniques are able to perform much better and generate more concise summaries. This paper proposes an efficient summarization system for patents using artificial intelligence, natural language processing and deep learning techniques to condense the knowledge and essential information from a patent document into a single summary that is easier to understand without any redundant formatting and difficult jargon.

Keywords: abstractive summarization, deep learning, natural language Processing, patent document

Procedia PDF Downloads 116

Authors: Austin Jiang, Richard M. Salmon, Nicholas W. Morrell, Wei Li

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BMP10 is highly expressed in the developing heart and plays essential roles in cardiogenesis. BMP10 deletion in mice results in embryonic lethality due to impaired cardiac development. In adults, BMP10 expression is restricted to the right atrium, though ventricular hypertrophy is accompanied by increased BMP10 expression in a rat hypertension model. However, reports of BMP10 activity in the circulation are inconclusive. In particular it is not known whether in vivo secreted BMP10 is active or whether additional factors are required to achieve its bioactivity. It has been shown that high-affinity binding of the BMP10 prodomain to the mature ligand inhibits BMP10 signaling activity in C2C12 cells, and it was proposed that prodomain-bound BMP10 (pBMP10) complex is latent. In this study, we demonstrated that the BMP10 prodomain did not inhibit BMP10 signaling activity in multiple endothelial cells, and that recombinant human pBMP10 complex, expressed in mammalian cells and purified under native conditions, was fully active. In addition, both BMP10 in human plasma and BMP10 secreted from the mouse right atrium were fully active. Finally, we confirmed that active BMP10 secreted from mouse right atrium was in the prodomain-bound form. Our data suggest that circulating BMP10 in adults is fully active and that the reported vascular quiescence function of BMP10 in vivo is due to the direct activity of pBMP10 and does not require an additional activation step. Moreover, being an active ligand, recombinant pBMP10 may have therapeutic potential as an endothelial-selective BMP ligand, in conditions characterized by loss of BMP9/10 signaling.

Keywords: bone morphogenetic protein 10 (BMP10), endothelial cell, signal transduction, transforming growth factor beta (TGF-B)

Procedia PDF Downloads 271

Authors: Mohammad H. Yarmohammadian, Reza Safdari, Mahmoud Keyvanara, Nahid Tavakoli

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One of the greatest challenges in disaster and emergencies is patient tracking. The concept of tracking has different denotations. One of the meanings refers to tracking patients’ physical locations and the other meaning refers to tracking patients ‘medical needs during emergency services. The main goal of patient tracking is to provide patient safety during disaster and emergencies and manage the flow of patient and information in different locations. In most of cases, there are not sufficient and accurate data regarding the number of injuries, medical conditions and their accommodation and transference. The objective of the present study is to survey on patient tracking issue in natural disaster and emergencies. Methods: This was a narrative study in which the population was E-Journals and the electronic database such as PubMed, Proquest, Science direct, Elsevier, etc. Data was gathered by Extraction Form. All data were analyzed via content analysis. Results: In many countries there is no appropriate and rapid method for tracking patients and transferring victims after the occurrence of incidents. The absence of reliable data of patients’ transference and accommodation, even in the initial hours and days after the occurrence of disasters, and coordination for appropriate resource allocation, have faced challenges for evaluating needs and services challenges. Currently, most of emergency services are based on paper systems, while these systems do not act appropriately in great disasters and incidents and this issue causes information loss. Conclusion: Patient tracking system should update the location of patients or evacuees and information related to their states. Patients’ information should be accessible for authorized users to continue their treatment, accommodation and transference. Also it should include timely information of patients’ location as soon as they arrive somewhere and leave therein such a way that health care professionals can be able to provide patients’ proper medical treatment.

Keywords: patient tracking, challenges, disaster, emergency

Procedia PDF Downloads 298

Authors: Raymond Ndhlovu, Jen Snowball

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It has long been known that the Cultural and Creative Industries (CCIs) have the potential to aid in physical, social and economic renewal and regeneration of towns and cities, hence their importance when dealing with regional development. The CCIs can act as a catalyst for activity and investment in an area because the ‘consumption’ of cultural activities will lead to the activities and use of other non-cultural activities, for example, hospitality development including restaurants and bars, as well as public transport. ‘Consumption’ of cultural activities also leads to employment creation, and diversification. However, CCIs tend to be clustered, especially around large cities. There is, moreover, a case for development of CCIs around smaller towns and cities, because they do not rely on high technology inputs, and long supply chains, and, their direct link to rural and isolated places makes them vital in regional development. However, there is currently little research on how to craft cultural policy for regions with smaller towns and cities. Using the Sarah Baartman District (SBDM) in South Africa as an example, this paper describes the process of developing cultural policy for a region that has potential, and existing, cultural clusters, but currently no one, coherent policy relating to CCI development. The SBDM was chosen as a case study because it has no large cities, but has some CCI clusters, and has identified them as potential drivers of local economic development. The process of developing cultural policy is discussed in stages: Identification of what resources are present; including human resources, soft and hard infrastructure; Identification of clusters; Analysis of CCI labour markets and ownership patterns; Opportunities and challenges from the point of view of CCIs and other key stakeholders; Alignment of regional policy aims with provincial and national policy objectives; and finally, design and implementation of a regional cultural policy.

Keywords: cultural and creative industries, economic impact, intrinsic value, regional development

Procedia PDF Downloads 227

Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab

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Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.

Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations

Procedia PDF Downloads 335

Authors: Kai-Jui Kou, Tzu-Ling Shen, Ying-Fang Wang

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The objectives of the present study are to characterize nanoparticles generated from the different working type in 3D printing room and the stack flue during 3D printing process. The studied laboratory (10.5 m× 7.2 m × 3.2 m) with a ventilation rate of 500 m³/H is installed a 3D metal printing machine. Direct-reading instrument of a scanning mobility particle sizer (SMPS, Model 3082, TSI Inc., St. Paul, MN, USA) was used to conduct static sampling for nanoparticle number concentration and particle size distribution measurements. The SMPS obtained particle number concentration at every 3 minutes, the diameter of the SMPS ranged from 11~372 nm when the aerosol and sheath flow rates were set at 0.6 and 6 L/min, respectively. The concentrations of background, printing process, clearing operation, and screening operation were performed in the laboratory. On the other hand, we also conducted nanoparticle measurement on the 3D printing machine's stack flue to understand its emission characteristics. Results show that the nanoparticles emitted from the different operation process were the same distribution in the form of the uni-modal with number median diameter (NMD) as approximately 28.3 nm to 29.6 nm. The number concentrations of nanoparticles were 2.55×10³ count/cm³ in laboratory background, 2.19×10³ count/cm³ during printing process, 2.29×10³ count/cm³ during clearing process, 3.05×10³ count/cm³ during screening process, 2.69×10³ count/cm³ in laboratory background after printing process, and 6.75×10³ outside laboratory, respectively. We found that there are no emission nanoparticles during the printing process. However, the number concentration of stack flue nanoparticles in the ongoing print is 1.13×10⁶ count/cm³, and that of the non-printing is 1.63×10⁴ count/cm³, with a NMD of 458 nm and 29.4 nm, respectively. It can be confirmed that the measured particle size belongs to easily penetrate the filter in theory during the printing process, even though the 3D printer has a high-efficiency filtration device. Therefore, it is recommended that the stack flue of the 3D printer would be equipped with an appropriate dust collection device to prevent the operators from exposing these hazardous particles.

Keywords: nanoparticle, particle emission, 3D printing, number concentration

Procedia PDF Downloads 173

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

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Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

Procedia PDF Downloads 219

Authors: Miroslava Rysová, Zdena Syrová, Tomáš Zajíc, Petr Exnar

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Currently, attention of tissue engineers has been attracted to novel nanofibrous materials having advanced properties and ability to mimic extracellular matrix (ECM) by structure which makes them interesting candidates for application in regenerative medicine as scaffolding and/or drug delivering material. Throughout the last decade, more than 200 synthetic and natural polymers have been successfully electrospun leading to the formation of nanofibres with a wide range of chemical, mechanical and degradation properties. In this family, inorganic nanofibres represent very specific group offering an opportunity to manufacture inert to body, well degradable and in properties tunable material. Aim of this work, was to reveal unique properties of silica (SiO2, CAS 7631-86-9) nanofibres and their potential in field of regenerative medicine. Silica nanofibres were prepared by sol-gel method from tetraethyl orthosilicate (TEOS, CAS 78-10-4) as a precursor and subsequently manufactured by needleless electrospinning on NanospiderTM device. Silica nanofibres thermally stabilized under 200°C were confirmed to be fully biodegradable and soluble in several simulated body fluids. In vitro cytotoxicity tests of eluate (ES ISO 10993-5:1999) and in direct contact (ES ISO 10993-5:2009) showed no toxicity - e.g. cell viabilities reached values exceeding 80%. Those results were obtained equally from two different cell lines (Vero, 3T3). Non-toxicity of silaca nanofibres´ eluate was additionally confirmed in real time by testing on xCelligence (ACEA Biosciences, Inc.) device. Both cell types also showed good adhesion to material. To conclude, all mentioned results lead to resumption that silica nanofibres have a potential as material for regenerative medicine which opens door to further research.

Keywords: cytotoxicity, electrospinning, nanofibres, silica, tissue engineering

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Authors: N. Valderrama, W. Albarracín, N. Algecira

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It was studied the effect of the inclusion of thyme and rosemary essential oils into chitosan films, as well as the microbiological and physical properties when storing chitosan film with and without the mentioned inclusion. The film forming solution was prepared by dissolving chitosan (2%, w/v), polysorbate 80 (4% w/w CH) and glycerol (16% w/w CH) in aqueous lactic acid solutions (control). The thyme (TEO) and rosemary (REO) essential oils (EOs) were included 1:1 w/w (EOs:CH) on their combination 50/50 (TEO:REO). The films were stored at temperatures of 5, 20, 33°C and a relative humidity of 75% during four weeks. The films with essential oil inclusion did not show an antimicrobial activity against strains. This behavior could be explained because the chitosan only inhibits the growth of microorganisms in direct contact with the active sites. However, the inhibition capacity of TEO was higher than the REO and a synergic effect between TEO:REO was found for S. enteritidis strains in the chitosan solution. Some physical properties were modified by the inclusion of essential oils. The addition of essential oils does not affect the mechanical properties (tensile strength, elongation at break, puncture deformation), the water solubility, the swelling index nor the DSC behavior. However, the essential oil inclusion can significantly decrease the thickness, the moisture content, and the L* value of films whereas the b* value increased due to molecular interactions between the polymeric matrix, the loosing of the structure, and the chemical modifications. On the other hand, the temperature and time of storage changed some physical properties on the chitosan films. This could have occurred because of chemical changes, such as swelling in the presence of high humidity air and the reacetylation of amino groups. In the majority of cases, properties such as moisture content, tensile strength, elongation at break, puncture deformation, a*, b*, chrome, ΔE increased whereas water resistance, swelling index, L*, and hue angle decreased.

Keywords: chitosan, food additives, modified films, polymers

Procedia PDF Downloads 362

Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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Authors: Seyed Reza Aghili, Tahereh Shokohi, Lotfollah Davoodi, Zahra Kashi, Azam Moslemi, Mahdi Abastabar, Iman Haghani, Sabah Mayahi, Asoudeh A.

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Introduction: In diabetic foot ulcers, if fungal agents such as Candida species penetrate into the cutaneous or depth of ulcer, can increase the degree of the wound and cause Candia infection and make it more difficult to heal. Material & Methods: A cross-sectional study was performed on 100 diabetic foot ulcer patients in 2020 in Sari, Iran. patient's data and wound grade were recorded in a questionnaire. Candida infection was diagnosed with direct microscopic examination and culture of samples. Colony-PCR molecular method was used for ITS region of DNA and then PCR-RFLP with Msp1 enzyme and using HWP1 specific gene to determine species of Candida agent. Results: Of 100 patients, the mean age 62.1 ± 10.8 years, 95% type 2 diabetes, 83%>10 years duration diabetes, 59% male, 66%> poor education level, 99% married, 52% rural, 95% neuropathic symptoms, 88% using antibiotics, 69%HbA1C >9%, and mean ulcer degree 2.6±1.05 were. Candida infection was seen in 13% of the deep tissue of the wound and 7% cutaneous around the wound. The predominant Candida isolated was C. parapsilosis (71.5%), C .albicans (14.3%). Fungal infections caused by mold fungi were not detected. There was a statistically significant relationship between yeast infection and gender, rural, HbA1C and ulcer degree. Conclusion: Mycological evaluations often are ignored. Candida parapsilosis is the most common infectious agent in these patients and may require specific treatment. Therefore, more attention of physicians to Candida infections particularly, early diagnosis and effective treatment can help faster recovery and prevent amputation.

Keywords: diabetic foot ulcer, candida infection, risk factors, c. parapsilosis

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Authors: C. Busuioc, S. I. Jinga, E. Pavel

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The continuous need of more non-volatile memories with a higher storage capacity, smaller dimensions and weight, as well as lower costs, has led to the exploration of optical lithography on active media, as well as patterned magnetic composites. In this context, optical lithography is a technique that can provide a significant decrease of the information bit size to the nanometric scale. However, there are some restrictions that arise from the need of breaking the optical diffraction limit. Major achievements have been obtained by employing a vitoceramic material as active medium and a laser beam operated at low power for the direct writing procedure. Thus, optical discs with ultra-high density were fabricated by a conventional melt-quenching method starting from analytical purity reagents. They were subsequently used for 3D recording based on their photosensitive features. Naturally, the next step consists in the elucidation of the composition and structure of the active centers, in correlation with the use of silver and rare-earth compounds for the synthesis of the optical supports. This has been accomplished by modern characterization methods, namely transmission electron microscopy coupled with selected area electron diffraction, scanning transmission electron microscopy and electron energy loss spectroscopy. The influence of laser diode parameters, silver concentration and fluorescent compounds formation on the writing process and final material properties was investigated. The results indicate performances in terms of capacity with two order of magnitude higher than other reported information storage systems. Moreover, the fluorescent photosensitive vitroceramics may be integrated in other applications which appeal to nanofabrication as the driving force in electronics and photonics fields.

Keywords: data storage, fluorescent compounds, laser writing, vitroceramics

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Authors: Muhammad Izzam Bin Mohd Khazar, Nur Adibah Binti Zainudin

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Islamic banking as one of the financial institutions is highly required to maintain a prudent approach to ensure that any financing given is able to generate income to their respective shareholders. As the default payment of customers is probably occurred in the financing, having a prudent approach in the recovery process is a must to ensure that financing losses are within acceptable limits. The objective of this research is to provide the best practice of recovery which is anticipated to benefit both bank and customers. This study will address arising issue on the current practice of recovery process and followed by providing humanising recovery solutions in the light of the Maqasid Shariah. The study identified main issues pertaining to Islamic recovery process which can be categorized into knowledge crisis, process issues, specific treatment cases and system issues. Knowledge crisis is related to direct parties including judges, solicitors and salesperson, while the recovery process issues include the process of issuance of reminder, foreclosure and repossession of asset. Furthermore, special treatment for particular cases also should be observed since different contracts in Islamic banking products will need different treatment. Finally, issues in the system used in the recovery process are still unresolved since the existing technology is still young in this area to embraced Islamic finance requirements and nature of calculation. In order to humanize the financial services in Islamic banking recovery process, we have highlighted four main recommendation to be implemented by Islamic Financial Institutions namely; 1) early deterrent by improving the awareness, 2) improvement of the internal process, 3) reward mechanism, and 4) creative penalty to provide awareness to all stakeholders.

Keywords: humanizing financial services, Islamic Finance, Maqasid Syariah, recovery process

Procedia PDF Downloads 190

Authors: Rony Tawk, Boutros Ghannam, Maroun Nemer

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Topology optimization technique presents a potential tool for the design and optimization of structures involved in mass and heat transfer. The method starts with an initial intermediate domain and should be able to progressively distribute the solid and the two fluids exchanging heat. The multi-objective function of the problem takes into account minimization of total pressure loss and maximization of heat transfer between solid and fluid subdomains. Existing methods account for the presence of only one fluid, while the actual work extends optimization distribution of solid and two different fluids. This requires to separate the channels of both fluids and to ensure a minimum solid thickness between them. This is done by adding a third objective function to the multi-objective optimization problem. This article uses density approach where each cell holds two local design parameters ranging from 0 to 1, where the combination of their extremums defines the presence of solid, cold fluid or hot fluid in this cell. Finite volume method is used for direct solver coupled with a discrete adjoint approach for sensitivity analysis and method of moving asymptotes for numerical optimization. Several examples are presented to show the ability of the method to find a trade-off between minimization of power dissipation and maximization of heat transfer while ensuring the separation and continuity of the channel of each fluid without crossing or mixing the fluids. The main conclusion is the possibility to find an optimal bi-fluid domain using topology optimization, defining a fluid to fluid heat exchanger device.

Keywords: topology optimization, density approach, bi-fluid domain, laminar steady state regime, fluid-to-fluid heat exchanger

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Authors: N. Attari, S. Amziane, M. Chemrouk

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A large number of old buildings have been identified as having potentially critical detailing to resist earthquakes. The main reinforcement of lap-spliced columns just above the joint region, discontinuous bottom beam reinforcement, and little or no joint transverse reinforcement are the most critical details of interior beam column joints in such buildings. This structural type constitutes a large share of the building stock, both in developed and developing countries, and hence it represents a substantial exposure. Direct observation of damaged structures, following the Algiers 2003 earthquake, has shown that damage occurs usually at the beam-column joints, with failure in bending or shear, depending on geometry and reinforcement distribution and type. While substantial literature exists for the design of concrete frame joints to withstand this type of failure, after the earthquake many structures were classified as slightly damaged and, being uneconomic to replace them, at least in the short term, suitable means of repairs of the beam column joint area are being studied. Furthermore; there exists a large number of buildings that need retrofitting of the joints before the next earthquake. The paper reports the results of the experimental programme, constituted of three beam-column reinforced concrete joints at a scale of one to three (1/3) tested under the effect of a pre-stressing axial load acting over the column. The beams were subjected at their ends to an alternate cyclic loading under displacement control to simulate a seismic action. Strain and cracking fields were monitored with the help a digital recording camera. Following the analysis of the results, a comparison can be made between the performances in terms of ductility, strength and mode of failure of the different strengthening solution considered.

Keywords: fibre reinforced polymers, joints, reinforced concrete, beam columns

Procedia PDF Downloads 409

Authors: M. Ali Lotfollahi Yaghin, M. Reza Bagerzadeh Karimi, Ali Rahmani, V. Sadeghi Balkanlou

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Structural engineers have long been trying to develop solutions using the full potential of its composing materials. Therefore, there is no doubt that the structural solution progress is directly related to an increase in materials science knowledge. These efforts in conjunction with up-to-date modern construction techniques have led to an extensive use of composite floors in large span structures. On the other hand, the competitive trends of the world market have long been forcing structural engineers to develop minimum weight and labour cost solutions. A direct consequence of this new design trend is a considerable increase in problems related to unwanted floor vibrations. For this reason, the structural floors systems become vulnerable to excessive vibrations produced by impacts such as human rhythmic activities. The main objective of this paper is to present an analysis methodology for the evaluation of the composite floors human comfort. This procedure takes into account a more realistic loading model developed to incorporate the dynamic effects induced by human walking. The investigated structural models were based on various composite floors, with main spans varying from 5 to 10 m. based on an extensive parametric study the composite floors dynamic response, in terms of peak accelerations, was obtained and compared to the limiting values proposed by several authors and design standards. This strategy was adopted to provide a more realistic evaluation for this type of structure when subjected to vibration due to human walking.

Keywords: vibration, resonance, composite floors, people’s rhythmic movement, dynamic analysis, Abaqus software

Procedia PDF Downloads 295

Authors: Lizinis Cassendra Frederick Dony, Dewi Binti Tajuddin, Jirom Jeremy Frederick Dony, Andrew Nicholas

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Unemployed graduates have become among the world major concerns lately. 70% of Malaysian graduates are jobless. Due to this concerns, this paper aims to identify major factors influencing Sabah minority ethnic favorable in the entrepreneurialism business interest. Hence, extensive introductory entrepreneurship syllabus clusters development beginning from the early childhood, primary, secondary and university students. This may induce interest appeal and to develop focus group of self-employment. The study focus on 7 indicators consist of demographic profiles variable (DP), social norms (SN), attitude (A), self-efficacy (SE) and business management skills (BMS) with reference to university students’ entrepreneurial intention. This study also partially mediates the relationship between product attractiveness (PA) and the minority ethnic entrepreneurialism business interest (MEEBI), by testifying their direct and indirect relationships. Hence, this study provides new perception towards improving the graduates’ characteristic, capabilities to exploit the business opportunities in the market. Furthermore, this paper will assess the relationship with the product attractiveness(PA) as a mediator. The study encompasses on the type of Sabah minority ethnic (ME) and nature of family own business (FOB) background with the 280 samples students in UiTM Sabah and UMS. The descriptive and random sampling method of research which revealed that majority of the respondents agreed that FOB and entrepreneurship education had positively influenced the ethnic students’ involvement in the entrepreneurial process and career development either full-time or part-time basis.

Keywords: Demographic profile (DP), soci, unemployed graduates, Malaysian minority

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Authors: Charles Amoo, Joshua New, Bill Eckman

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Buildings in the United States account for a significant proportion of energy consumption and greenhouse gas (GHG) emissions, and this trend is expected to continue as well as rise in the near future. Low-income households, in particular, bear a disproportionate burden of high building energy consumption and spending due to high energy costs. Energy efficiency improvements need to reach an average of 4% per year in this decade in order to meet global net zero emissions target by 2050, but less than 1 % of U.S. buildings are improved each year. The government has recognized the importance of technology in addressing this issue, and energy efficiency programs have been developed to tackle the problem. The Weatherization Assistance Program (WAP), the largest residential whole-house energy efficiency program in the U.S., is specifically designed to reduce energy costs for low-income households. Under the WAP, energy auditors must follow specific audit procedures and use Department of Energy (DOE) approved energy audit tools or software. This article proposes an expanded framework of factors that should be considered in energy audit software that is approved for use in energy efficiency programs, particularly for low-income households. The framework includes more than 50 factors organized under 14 assessment criteria and can be used to qualitatively and quantitatively score different energy audit software to determine their suitability for specific energy efficiency programs. While the tool can be useful for developers to build new tools and improve existing software, as well as for energy efficiency program administrators to approve or certify tools for use, there are limitations to the model, such as the lack of flexibility that allows continuous scoring to accommodate variability and subjectivity. These limitations can be addressed by using aggregate scores of each criterion as weights that could be combined with value function and direct rating scores in a multicriteria decision analysis for a more flexible scoring.

Keywords: buildings, energy efficiency, energy audit, software

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Authors: Supriya Majumder, Pabitra Banik

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Fates of Arsenic (As) on the soil-plant environment belong to the critical emerging issue, which in turn to appraises the threatening implications of a human health risk — assessing the dynamics of As in soil solid components are likely to impose its potential availability towards plant uptake. In the present context, we introduced an improved Sequential Extraction Procedure (SEP) questioning to identify solid-phase speciation of As in paddy soil under variable soil environmental conditions during two consecutive seasons of rice cultivation practices. We coupled gradients of water management practices with the addition of fertilizer amendments to assess the changes in a partition of As through a field experimental study during monsoon and post-monsoon season using two rice cultivars. Water management regimes were varied based on the methods of cultivation of rice by Conventional (waterlogged) vis-a-vis System of Rice Intensification-SRI (saturated). Fertilizer amendment through the nutrient treatment of absolute control, NPK-RD, NPK-RD + Calcium silicate, NPK-RD + Ferrous sulfate, Farmyard manure (FYM), FYM + Calcium silicate, FYM + Ferrous sulfate, Vermicompost (VC), VC + Calcium silicate, VC + Ferrous sulfate were selected to construct the study. After harvest, soil samples were sequentially extracted to estimate partition of As among the different fractions such as: exchangeable (F1), specifically sorbed (F2), As bound to amorphous Fe oxides (F3), crystalline Fe oxides (F4), organic matter (F5) and residual phase (F6). Results showed that the major proportions of As were found in F3, F4 and F6, whereas F1 exhibited the lowest proportion of total soil As. Among the nutrient treatment mediated changes on As fractions, the application of organic manure and ferrous sulfate were significantly found to restrict the release of As from exchangeable phase. Meanwhile, conventional practice produced much higher release of As from F1 as compared to SRI, which may substantially increase the environmental risk. In contrast, SRI practice was found to retain a significantly higher proportion of As in F2, F3, and F4 phase resulting restricted mobilization of As. This was critically reflected towards rice grain As bioavailability where the reduction in grain As concentration of 33% and 55% in SRI concerning conventional treatment (p <0.05) during monsoon and post-monsoon season respectively. Also, prediction assay for rice grain As bioavailability based on the linear regression model was performed. Results demonstrated that rice grain As concentration was positively correlated with As concentration in F1 and negatively correlated with F2, F3, and F4 with a satisfactory level of variation being explained (p <0.001). Finally, we conclude that F1, F2, F3 and F4 are the major soil. As fractions critically may govern the potential availability of As in soil and suggest that rice cultivation with the SRI treatment is particularly at less risk of As availability in soil. Such exhaustive information may be useful for adopting certain management practices for rice grown in contaminated soil concerning to the environmental issues in particular.

Keywords: arsenic, fractionation, paddy soil, potential availability

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Authors: Peng Hou

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Natural products produced from marine bacteria serve as an immense reservoir for anti-infective drugs and therapeutic agents. Nowadays, heterologous expression of gene clusters of interests has been widely adopted as an effective strategy for natural product discovery. Briefly, the heterologous expression flowchart would be: biosynthetic gene cluster identification, pathway construction and expression, and product detection. However, gene cluster capture using traditional Transformation-associated recombination (TAR) protocol is low-efficient (0.5% positive colony rate). To make things worse, most of these putative new natural products are only predicted by bioinformatics analysis such as antiSMASH, and their corresponding natural products biosynthetic pathways are either not expressed or expressed at very low levels under laboratory conditions. Those setbacks have inspired us to focus on seeking new technologies to efficiently edit and refractor of biosynthetic gene clusters. Recently, two cutting-edge techniques have attracted our attention - the CRISPR-Cas9 and Gibson Assembly. By now, we have tried to pretreat Brevibacillus laterosporus strain genomic DNA with CRISPR-Cas9 nucleases that specifically generated breaks near the gene cluster of interest. This trial resulted in an increase in the efficiency of gene cluster capture (9%). Moreover, using Gibson Assembly by adding/deleting certain operon and tailoring enzymes regardless of end compatibility, the silent construct (~80kb) has been successfully refactored into an active one, yielded a series of analogs expected. With the appearances of the novel molecular tools, we are confident to believe that development of a high throughput mature pipeline for DNA assembly, transformation, product isolation and identification would no longer be a daydream for marine natural product discovery.

Keywords: biosynthesis, CRISPR-Cas9, DNA assembly, refactor, TAR cloning

Procedia PDF Downloads 273