Search results for: asset specificity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 990

Search results for: asset specificity

300 Optimization and Kinetic Analysis of the Enzymatic Hydrolysis of Oil Palm Empty Fruit Bunch To Xylose Using Crude Xylanase from Trichoderma Viride ITB CC L.67

Authors: Efri Mardawati, Ronny Purwadi, Made Tri Ari Penia Kresnowati, Tjandra Setiadi

Abstract:

EFB are mainly composed of cellulose (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). The palm oil empty fruit bunches (EFB) is the lignosellulosic waste from crude palm oil industries mainly compose of (≈ 43%), hemicellulose (≈ 23%) and lignin (≈20%). Xylan, a polymer made of pentose sugar xylose and the most abundant component of hemicellulose in plant cell wall. Further xylose can be used as a raw material for production of a wide variety of chemicals such as xylitol, which is extensively used in food, pharmaceutical and thin coating applications. Currently, xylose is mostly produced from xylan via chemical hydrolysis processes. However, these processes are normally conducted at a high temperature and pressure, which is costly, and the required downstream processes are relatively complex. As an alternative method, enzymatic hydrolysis of xylan to xylose offers an environmentally friendly biotechnological process, which is performed at ambient temperature and pressure with high specificity and at low cost. This process is catalysed by xylanolytic enzymes that can be produced by some fungal species such as Aspergillus niger, Penicillium crysogenum, Tricoderma reseei, etc. Fungal that will be used to produce crude xylanase enzyme in this study is T. Viride ITB CC L.67. It is the purposes of this research to study the influence of pretreatment of EFB for the enzymatic hydrolysis process, optimation of temperature and pH of the hydrolysis process, the influence of substrate and enzyme concentration to the enzymatic hydrolysis process, the dynamics of hydrolysis process and followingly to study the kinetics of this process. Xylose as the product of enzymatic hydrolysis process analyzed by HPLC. The results show that the thermal pretreatment of EFB enhance the enzymatic hydrolysis process. The enzymatic hydrolysis can be well approached by the Michaelis Menten kinetic model, and kinetic parameters are obtained from experimental data.

Keywords: oil palm empty fruit bunches (EFB), xylose, enzymatic hydrolysis, kinetic modelling

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299 Consequence of Multi-Templating of Closely Related Structural Analogues on a Chitosan-Methacryllic Acid Molecularly Imprinted Polymer Matrix-Thermal and Chromatographic Traits

Authors: O.Ofoegbu, S. Roongnapa, A.N. Eboatu

Abstract:

Most polluted environments, most challengingly, aerosol types, contain a cocktail of different toxicants. Multi-templating of matrices have been the recent target by researchers in a bid to solving complex mixed-toxicant challenges using single or common remediation systems. This investigation looks at the effect of such multi-templated system vis-a-vis the synthesis by non-covalent interaction, of a molecularly imprinted polymer architecture using nicotine and its structural analogue Phenylalanine amide individually and, in the blend, (50:50), as template materials in a Chitosan-Methacrylic acid functional monomer matrix. The temperature for polymerization is 60OC and time for polymerization, 12hrs (water bath heating), 4mins for (microwave heating). The characteristic thermal properties of the molecularly imprinted materials are investigated using Simultaneous Thermal Analysis (STA) profiling, while the absorption and separation efficiencies based on the relative retention times and peak areas of templates were studied amongst other properties. Transmission Electron Microscopy (TEM) results obtained, show the creation of heterogeneous nanocavities, regardless, the introduction of Caffeine a close structural analogue presented near-zero perfusion. This confirms the selectivity and specificity of the templated polymers despite its dual-templated nature. The STA results presented the materials as having decomposition temperatures above 250OC and a relative loss in mass of less than19% over a period within 50mins of heating. Consequent to this outcome, multi-templated systems can be fabricated to sequester specifically and selectively targeted toxicants in a mixed toxicant populated system effectively.

Keywords: chitosan, dual-templated, methacrylic acid, mixed-toxicants, molecularly-imprinted-polymer

Procedia PDF Downloads 103
298 Prevalence and Comparison for Detection Methods of Candida Species in Vaginal Specimens from Pregnant and Non-Pregnant Saudi Women

Authors: Yazeed Al-Sheikh

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Pregnancy represents a risk factor in the occurrence of vulvovaginal candidiasis. To investigate the prevalence rate of vaginal carriage of Candida species in Saudi pregnant and non-pregnant women, high vaginal swab (HVS) specimens (707) were examined by direct microscopy (10% KOH and Giemsa staining) and parallel cultured on Sabouraud Dextrose Agar (SDA) as well as on “CHROM agar Candida” medium. As expected, Candida-positive cultures were frequently observed in pregnant-test group (24%) than in non-pregnant group (17%). The frequency of culture positive was correlated to pregnancy (P=0.047), parity (P=0.001), use of contraceptive (P=0.146), or antibiotics (P=0.128), and diabetic-patients (P < 0.0001). Out of 707 HVS examined specimens, 157 specimens were yeast-positive culture (22%) on Sabouraud Dextrose Agar or “CHROM agar Candida”. In comparison, the sensitivities of the direct 10% KOH and the Giemsa stain microscopic examination methods were 84% (132/157) and 95% (149/157) respectively but both with 100% specificity. As for the identity of recovered 157 yeast isolates, based on API 20C biotype carbohydrate assimilation, germ tube and chlamydospore formation, C. albicansand C. glabrata constitute 80.3 and 12.7% respectively. Rates of C. tropicalis, C. kefyr, C. famata or C. utilis were 2.6, 1.3, and 0.6% respectively. Sachromyces cerevisiae and Rhodotorula mucilaginosa yeasts were also encountered at a frequency of 1.3 and 0.6% respectively. Finally, among all recovered 157 yeast-isolates, strains resistant to ketoconazole were not detected, whereas 5% of the C. albicans and as high as 55% of the non-albicans yeast isolates (majority C. glabrata) showed resistance to fluconazole. Our findings may prove helpful for continuous determination of the existing vaginal candidiasis causative species during pregnancy, its lab-diagnosis and/or control and possible measures to minimize the incidence of the disease-associated pre-term delivery.

Keywords: vaginal candidiasis, Candida spp., pregnancy, risk factors, API 20C-yeast biotypes, giemsa stain, antifungal agents

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297 Transcriptome Analysis Reveals Role of Long Non-Coding RNA NEAT1 in Dengue Patients

Authors: Abhaydeep Pandey, Shweta Shukla, Saptamita Goswami, Bhaswati Bandyopadhyay, Vishnampettai Ramachandran, Sudhanshu Vrati, Arup Banerjee

Abstract:

Background: Long non-coding RNAs (lncRNAs) are the important regulators of gene expression and play important role in viral replication and disease progression. The role of lncRNA genes in the pathogenesis of Dengue virus-mediated pathogenesis is currently unknown. Methods: To gain additional insights, we utilized an unbiased RNA sequencing followed by in silico analysis approach to identify the differentially expressed lncRNA and genes that are associated with dengue disease progression. Further, we focused our study on lncRNAs NEAT1 (Nuclear Paraspeckle Assembly Transcript 1) as it was found to be differentially expressed in PBMC of dengue infected patients. Results: The expression of lncRNAs NEAT1, as compared to dengue infection (DI), was significantly down-regulated as the patients developed the complication. Moreover, pairwise analysis on follow up patients confirmed that suppression of NEAT1 expression was associated with rapid fall in platelet count in dengue infected patients. Severe dengue patients (DS) (n=18; platelet count < 20K) when recovered from infection showing high NEAT1 expression as it observed in healthy donors. By co-expression network analysis and subsequent validation, we revealed that coding gene; IFI27 expression was significantly up-regulated in severe dengue cases and negatively correlated with NEAT1 expression. To discriminate DI from dengue severe, receiver operating characteristic (ROC) curve was calculated. It revealed sensitivity and specificity of 100% (95%CI: 85.69 – 97.22) and area under the curve (AUC) = 0.97 for NEAT1. Conclusions: Altogether, our first observations demonstrate that monitoring NEAT1and IFI27 expression in dengue patients could be useful in understanding dengue virus-induced disease progression and may be involved in pathophysiological processes.

Keywords: dengue, lncRNA, NEAT1, transcriptome

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296 Anti-Parasite Targeting with Amino Acid-Capped Nanoparticles Modulates Multiple Cellular Processes in Host

Authors: Oluyomi Stephen Adeyemi, Kentaro Kato

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Toxoplasma gondii is the etiological agent of toxoplasmosis, a common parasitic disease capable of infecting a range of hosts, including nearly one-third of the human population. Current treatment options for toxoplasmosis patients are limited. In consequence, toxoplasmosis represents a large global burden that is further enhanced by the shortcomings of the current therapeutic options. These factors underscore the need for better anti-T. gondii agents and/or new treatment approach. In the present study, we sought to find out whether preparing and capping nanoparticles (NPs) in amino acids, would enhance specificity toward the parasite versus the host cell. The selection of amino acids was premised on the fact that T. gondii is auxotrophic for some amino acids. The amino acid-nanoparticles (amino-NPs) were synthesized, purified and characterized following established protocols. Next, we tested to determine the anti-T. gondii activity of the amino-NPs using in vitro experimental model of infection. Overall, our data show evidence that supports enhanced and excellent selective action against the parasite versus the host cells by amino-NPs. The findings are promising and provide additional support that warrants exploring the prospects of NPs as alternative anti-parasite agents. In addition, the anti-parasite action by amino-NPs indicates that nutritional requirement of parasite may represent a viable target in the development of better alternative anti-parasite agents. Furthermore, data suggest the anti-parasite mechanism of the amino-NPs involves multiple cellular processes including the production of reactive oxygen species (ROS), modulation of hypoxia-inducing factor-1 alpha (HIF-1α) as well as the activation of kynurenine pathway. Taken together, findings highlight further, the prospects of NPs as alternative source of anti-parasite agents.

Keywords: drug discovery, infectious diseases, mode of action, nanomedicine

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295 Development of Broad Spectrum Nitrilase Biocatalysts and Bioprocesses for Nitrile Biotransformation

Authors: Avinash Vellore Sunder, Shikha Shah, Pramod P. Wangikar

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The enzymatic conversion of nitriles to carboxylic acids by nitrilases has gained significance in the green synthesis of several pharmaceutical precursors and fine chemicals. While nitrilases have been characterized from different sources, the industrial application requires the identification of nitrilases that possess higher substrate tolerance, wider specificity and better thermostability, along with the development of an efficient bioprocess for producing large amounts of nitrilase. To produce large amounts of nitrilase, we developed a fed-batch fermentation process on defined media for the high cell density cultivation of E. coli cells expressing the well-studied nitrilase from Alcaligenes fecalis. A DO-stat feeding approach was employed combined with an optimized post-induction strategy to achieve nitrilase titer of 2.5*105 U/l and 78 g/l dry cell weight. We also identified 16 novel nitrilase sequences from genome mining and analysis of substrate binding residues. The nitrilases were expressed in E. coli and their biocatalytic potential was evaluated on a panel of 22 industrially relevant nitrile substrates using high-throughput screening and HPLC analysis. Nine nitrilases were identified to exhibit high activity on structurally diverse nitriles including aliphatic and aromatic dinitriles, heterocyclic, -hydroxy and -keto nitriles. With fed-batch biotransformation, whole-cell Zobelia galactanivorans nitrilase achieved yields of 2.4 M nicotinic acid and 1.8 M isonicotinic acid from 3-cyanopyridine and 4-cyanopyridine respectively within 5 h, while Cupravidus necator nitrilase enantioselectively converted 740 mM mandelonitrile to (R)–mandelic acid. The nitrilase from Achromobacter insolitus could hydrolyze 542 mM iminodiacetonitrile in 1 h. The availability of highly active nitrilases along with bioprocesses for enzyme production expands the toolbox for industrial biocatalysis.

Keywords: biocatalysis, isonicotinic acid, iminodiacetic acid, mandelic acid, nitrilase

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294 A Continuous Real-Time Analytic for Predicting Instability in Acute Care Rapid Response Team Activations

Authors: Ashwin Belle, Bryce Benson, Mark Salamango, Fadi Islim, Rodney Daniels, Kevin Ward

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A reliable, real-time, and non-invasive system that can identify patients at risk for hemodynamic instability is needed to aid clinicians in their efforts to anticipate patient deterioration and initiate early interventions. The purpose of this pilot study was to explore the clinical capabilities of a real-time analytic from a single lead of an electrocardiograph to correctly distinguish between rapid response team (RRT) activations due to hemodynamic (H-RRT) and non-hemodynamic (NH-RRT) causes, as well as predict H-RRT cases with actionable lead times. The study consisted of a single center, retrospective cohort of 21 patients with RRT activations from step-down and telemetry units. Through electronic health record review and blinded to the analytic’s output, each patient was categorized by clinicians into H-RRT and NH-RRT cases. The analytic output and the categorization were compared. The prediction lead time prior to the RRT call was calculated. The analytic correctly distinguished between H-RRT and NH-RRT cases with 100% accuracy, demonstrating 100% positive and negative predictive values, and 100% sensitivity and specificity. In H-RRT cases, the analytic detected hemodynamic deterioration with a median lead time of 9.5 hours prior to the RRT call (range 14 minutes to 52 hours). The study demonstrates that an electrocardiogram (ECG) based analytic has the potential for providing clinical decision and monitoring support for caregivers to identify at risk patients within a clinically relevant timeframe allowing for increased vigilance and early interventional support to reduce the chances of continued patient deterioration.

Keywords: critical care, early warning systems, emergency medicine, heart rate variability, hemodynamic instability, rapid response team

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293 The Right to a Fair Trial in French and Spanish Constitutional Law

Authors: Chloe Fauchon

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In Europe, the right to a fair trial is enshrined in the European Convention on Human Rights, signed in 1950, in its famous Article 6, and, in the field of the European Union, in Article 47 of the Charter of Fundamental Rights, binding since 2009. The right to a fair trial is, therefore, a fundamental right protected by all the relevant treaties. The right to a fair trial is an "umbrella right" which encompasses various sub-rights and principles. Although this right applies in all the proceedings, it gets a special relevance in criminal matters and, particularly, regarding the defendant. In criminal proceedings, the parties are not equal: the accusation is represented by a State-organ, with specific prerogatives, and the defense does not benefit from these specific powers and is often inexperienced in criminal law. Equality of arms, and consequently the right to a fair trial, needs some specific mechanisms to be effective in criminal proceedings. For instance, the defendant benefits from some procedural rights, such as the right to a lawyer, the right to be informed of the charges against them, the right to confront witnesses, and so on. These rights aim to give the defendant the tools to dispute the accusation. The role of the defense is, therefore, very important in criminal matters to avoid unjustified convictions. This specificity of criminal matters justifies that the focus will be put on them during this study. Then this paper will also focus on French and Spanish legal orders. Indeed, if the European Court and Convention on Human Rights are the most famous instruments to protect the right to a fair trial, this right is also guaranteed at a constitutional level in European national legal orders in Europe. However, this enshrinement differs from one country to the other: for instance, in Spain, the right to a fair trial is protected explicitly by the 1978 constitutional text, whereas, in France, it is more of a case-law construction. Nevertheless, this difference between both legal orders does not imply huge variations in the substantive aspect of the right to a fair trial. This can be specifically explained by the submission of both States to the European Convention on Human Rights. This work aims to show that, although the French and Spanish legal orders differ in the way they protect the right to a fair trial, this right eventually has the same substantive meaning in both legal orders.

Keywords: right to a fair trial, constitutional law, French law, Spanish law, European Court of Human Rights

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292 The Determinants of Enterprise Risk Management: Literature Review, and Future Research

Authors: Sylvester S. Horvey, Jones Mensah

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The growing complexities and dynamics in the business environment have led to a new approach to risk management, known as enterprise risk management (ERM). ERM is a system and an approach to managing the risks of an organization in an integrated manner to achieve the corporate goals and strategic objectives. Regardless of the diversities in the business environment, ERM has become an essential factor in managing individual and business risks because ERM is believed to enhance shareholder value and firm growth. Despite the growing number of literature on ERM, the question about what factors drives ERM remains limited. This study provides a comprehensive literature review of the main factors that contribute to ERM implementation. Google Scholar was the leading search engine used to identify empirical literature, and the review spanned between 2000 and 2020. Articles published in Scimago journal ranking and Scopus were examined. Thirteen firm characteristics and sixteen articles were considered for the empirical review. Most empirical studies agreed that firm size, institutional ownership, industry type, auditor type, industrial diversification, earnings volatility, stock price volatility, and internal auditor had a positive relationship with ERM adoption, whereas firm size, institutional ownership, auditor type, and type of industry were mostly seen be statistically significant. Other factors such as financial leverage, profitability, asset opacity, international diversification, and firm complexity revealed an inconclusive result. The growing literature on ERM is not without limitations; hence, this study suggests that further research should examine ERM determinants within a new geographical context while considering a new and robust way of measuring ERM rather than relying on a simple proxy (dummy) for ERM measurement. Other firm characteristics such as organizational culture and context, corporate scandals and losses, and governance could be considered determinants of ERM adoption.

Keywords: enterprise risk management, determinants, ERM adoption, literature review

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291 Capacity Oversizing for Infrastructure Sharing Synergies: A Game Theoretic Analysis

Authors: Robin Molinier

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Industrial symbiosis (I.S) rely on two basic modes of cooperation between organizations that are infrastructure/service sharing and resource substitution (the use of waste materials, fatal energy and recirculated utilities for production). The former consists in the intensification of use of an asset and thus requires to compare the incremental investment cost to be incurred and the stand-alone cost faced by each potential participant to satisfy its own requirements. In order to investigate the way such a cooperation mode can be implemented we formulate a game theoretic model integrating the grassroot investment decision and the ex-post access pricing problem. In the first period two actors set cooperatively (resp. non-cooperatively) a level of common (resp. individual) infrastructure capacity oversizing to attract ex-post a potential entrant with a plug-and-play offer (available capacity, tariff). The entrant’s requirement is randomly distributed and known only after investments took place. Capacity cost exhibits sub-additive property so that there is room for profitable overcapacity setting in the first period under some conditions that we derive. The entrant willingness-to-pay for the access to the infrastructure is driven by both her standalone cost and the complement cost to be incurred in case she chooses to access an infrastructure whose the available capacity is lower than her requirement level. The expected complement cost function is thus derived, and we show that it is decreasing, convex and shaped by the entrant’s requirements distribution function. For both uniform and triangular distributions optimal capacity level is obtained in the cooperative setting and equilibrium levels are determined in the non-cooperative case. Regarding the latter, we show that competition is deterred by the first period investor with the highest requirement level. Using the non-cooperative game outcomes which gives lower bounds for the profit sharing problem in the cooperative one we solve the whole game and describe situations supporting sharing agreements.

Keywords: capacity, cooperation, industrial symbiosis, pricing

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290 The Construction of Women’s Leadership in the Swedish Armed Forces in the Context of the Women, Peace and Security Agenda

Authors: Sofia Sutera

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Despite the introduction of the Women, Peace, and Security (WPS) Agenda in 2000, thanks to the UNSCR 1325 and subsequent resolutions, and the clear stance of the UN towards the support of increased participation of women in peace and security processes, women’s leadership in this context remains very low. Considering specifically the framework of peacekeeping operations, the aim of this paper is to analyze the way women’s leadership is constructed in the Swedish Armed Forces (SAF). In the context of the WPS Agenda, Sweden has been chosen as a case study because of the relevance of its singular feminist policies (the statement in 2014 from Wallström, previous and current Minister for Foreign Affairs and Deputy Prime Minister, that Sweden is pursuing a feminist foreign policy is a clear example). Moreover, the SAF adopted in 2016 the Handbok Gender. This policy addresses explicitly the gender perspective embraced by the Swedish military institution, a sui-generis organization even in the Scandinavian reality. Indeed, the SAF has assumed a clear commitment to represent its institution as gender aware and gender equal. The theoretical perspective utilized in this research, which focuses specifically on women, is feminism and particularly a feminist constructivist approach, with an institutional focus on the military institution, has been chosen. Taking into account the specificity of the feminist research, the above-mentioned gender policy has been examined by means of a critical discourse analysis (CDA) whose main aim is to investigate the social structures of discourse and the power relationships inherent to it. Thus, CDA appears to be quite relevant in order to understand the construction of women’s leadership in the Handbok Gender. Nevertheless, even in a country which officially identifies as feminist and which is characterized by a peculiar military institution, the conclusions of this analysis revealed that women’s leadership in peacekeeping operations remains very low.

Keywords: feminism, peacekeeping operations, swedish armed forces, UNSCR 1325, women's leadership, WPS agenda

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289 In Exile but Not at Peace: An Ethnography among Rwandan Army Deserters in South Africa

Authors: Florence Ncube

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This paper examines the military and post-military experiences of soldiers who deserted from the Rwanda Defence Force (RDF) and tried to make a living in South Africa. Because they are deserters, they try to hide their military identity, yet it is simultaneously somewhat coercively ascribed to them by the Rwandan state and can put them in potential danger. The paper attends to the constructions, experiences, practices, and subjective understanding of the deserters’ being in exile to examine how, under circumstances of perceived threat, these men navigate real or perceived state-sponsored surveillance and threat in non-military settings in South Africa where they have become potential political and disciplinary targets. To make sense of the deserters’ experiences in these circumstances, the paper stitches together a number of useful theoretical concepts, including Bourdieu’s (1992) theory of practice and Vigh’s (2009; 2018) concept of social navigation because no single approach can coherently analyze the specificity of this study. Conventional post-military literature privileges an understanding of army desertion as a malignancy and somewhat problematic. Little is known about the military and post-military experiences of deserters who believe that army desertion is in fact a building block towards achieving subjective peace, even in the context of exile. The paper argues that the presence of Rwandan state agents in South Africa strips the context of the exile of its capacity to provide the deserters with peace, safety, and security. This paper recenters army desertion in analyses of militarism, soldiering, and transition in African contexts and complicates commonsense understandings of army desertion which assume that it is entirely problematic. This paper is drawn from an ethnography conducted among 30 junior-rank Rwandan army deserters exiled in Johannesburg and Cape Town. The researcher employed life histories, in-depth interviews, and deep hangouts to collect data.

Keywords: army deserter, military, identity, exile, peacebuilding, South Africa

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288 Standard Protocol Selection for Acquisition of Breast Thermogram in Perspective of Early Breast Cancer Detection

Authors: Mrinal Kanti Bhowmik, Usha Rani Gogoi Jr., Anjan Kumar Ghosh, Debotosh Bhattacharjee

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In the last few decades, breast thermography has achieved an average sensitivity and specificity of 90% for breast tumor detection. Breast thermography is a non-invasive, cost-effective, painless and radiation-free breast imaging modality which makes a significant contribution to the evaluation and diagnosis of patients, suspected of having breast cancer. An abnormal breast thermogram may indicate significant biological risk for the existence or the development of breast tumors. Breast thermography can detect a breast tumor, when the tumor is in its early stage or when the tumor is in a dense breast. The infrared breast thermography is very sensitive to environmental changes for which acquisition of breast thermography should be performed under strictly controlled conditions by undergoing some standard protocols. Several factors like air, temperature, humidity, etc. are there to be considered for characterizing thermal images as an imperative tool for detecting breast cancer. A detailed study of various breast thermogram acquisition protocols adopted by different researchers in their research work is provided here in this paper. After going through a rigorous study of different breast thermogram acquisition protocols, a new standard breast thermography acquisition setup is proposed here in this paper for proper and accurate capturing of the breast thermograms. The proposed breast thermogram acquisition setup is being built in the Radiology Department, Agartala Government Medical College (AGMC), Govt. of Tripura, Tripura, India. The breast thermograms are captured using FLIR T650sc thermal camera with the thermal sensitivity of 20 mK at 30 degree C. The paper is an attempt to highlight the importance of different critical parameters of breast thermography like different thermography views, patient preparation protocols, acquisition room requirements, acquisition system requirements, etc. This paper makes an important contribution by providing a detailed survey and a new efficient approach on breast thermogram capturing.

Keywords: acquisition protocol, breast cancer, breast thermography, infrared thermography

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287 LTF Expression Profiling Which is Essential for Cancer Cell Proliferation and Metastasis, Correlating with Clinical Features, as Well as Early Stages of Breast Cancer

Authors: Azar Heidarizadi, Mahdieh Salimi, Hossein Mozdarani

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Introduction: As a complex disease, breast cancer results from several genetic and epigenetic changes. Lactoferrin, a member of the transferrin family, is reported to have a number of biological functions, including DNA synthesis, immune responses, iron transport, etc., any of which could play a role in tumor progression. The aim of this study was to investigate the bioinformatics data and experimental assay to find the pattern of promoter methylation and gene expression of LTF in breast cancer in order to study its potential role in cancer management. Material and Methods: In order to evaluate the methylation status of the LTF promoter, we studied the MS-PCR and Real-Time PCR on samples from patients with breast cancer and normal cases. 67 patient samples were conducted for this study, including tumoral, plasma, and normal tissue adjacent samples, as well as 30 plasma from normal cases and 10 tissue breast reduction cases. Subsequently, bioinformatics analyses such as cBioPortal databases, string, and genomatix were conducted to disclose the prognostic value of LTF in breast cancer progression. Results: The analysis of LTF expression showed an inverse relationship between the expression level of LTF and the stages of tissues of breast cancer patients (p<0.01). In fact, stages 1 and 2 had a high expression in LTF, while, in stages 3 and 4, a significant reduction was observable (p < 0.0001). LTF expression frequently alters with a decrease in the expression in ER⁺, PR⁺, and HER2⁺ patients (P < 0.01) and an increase in the expression in the TNBC, LN¯, ER¯, and PR- patients (P < 0.001). Also, LTF expression is significantly associated with metastasis and lymph node involvement factors (P < 0.0001). The sensitivity and specificity of LTF were detected, respectively. A negative correlation was detected between the results of level expression and methylation of the LTF promoter. Conclusions: The altered expression of LTF observed in breast cancer patients could be considered as a promotion in cell proliferation and metastasis even in the early stages of cancer.

Keywords: LTF, expression, methylation, breast cancer

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286 Pathogenic Effects of IgG and IgM Apoptotic Cell-Reactive Monoclonal Auto-Antibodies on Innate and Adaptive Immunity in Lupus

Authors: Monika Malik, Pooja Arora, Ruchi Sachdeva, Vishnampettai G. Ramachandran, Rahul Pal

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Apoptotic debris is believed to be the antigenic trigger in lupus. Whether such debris and autoantibodies induced in lupus-prone mice which specifically recognize its constituents can mediate differential effects on innate and humoral responses in such mice was assessed. The influence of apoptotic blebs and apoptotic cell-reactive monoclonal antibodies on phenotypic markers expressed on bone marrow-derived dendritic cells (BMDCs) and secreted cytokines were evaluated. Sera from lupus-prone and healthy mice immunized with the antibodies were analyzed for anti-self reactivity. Apoptotic blebs, as well as somatically-mutated IgG and non-mutated IgM apoptotic-cell reactive monoclonal antibodies, induced the preferential maturation of BMDCs derived from lupus-prone mice relative to BMDCs derived from healthy mice; antibody specificity and cell genotype both influenced the secretion of inflammatory cytokines. Immunization of lupus-prone mice with IgM and IgG antibodies led to hypergammaglobulinemia; elicited antibodies were self-reactive, and exhibited enhanced recognition of lupus-associated autoantigens (dsDNA, Ro60, RNP68, and Sm) in comparison with adjuvant-induced sera. While ‘natural’ IgM antibodies are believed to contribute to immune homeostasis, this study reveals that apoptotic cell-reactive IgM antibodies can promote inflammation and drive anti-self responses in lupus. Only in lupus-prone mice did immunization with IgG auto-antibodies enhance the kinetics of humoral anti-self responses, resulting in advanced-onset glomerulosclerosis. This study reveals that preferential innate and humoral recognition of the products of cell death in an autoimmune milieu influences the indices associated with lupus pathology.

Keywords: antigen spreading, apoptotic cell-reactive pathogenic IgG, and IgM autoantibodies, glomerulosclerosis, lupus

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285 Detection of Bcl2 Polymorphism in Patient with Hepatocellular carcinoma

Authors: Mohamed Abdel-Hamid, Olfat Gamil Shaker, Doha El-Sayed Ellakwa, Eman Fathy Abdel-Maksoud

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Introduction: Despite advances in the knowledge of the molecular virology of hepatitis C virus (HCV), the mechanisms of hepatocellular injury in HCV infection are not completely understood. Hepatitis C viral infection (HCV) influences the susceptibility to apoptosis. This could lead to insufficient antiviral immune response and persistent viral infection. Aim of this study: was to examine whether BCL-2 gene polymorphism at codon 43 (+127G/A or Ala43Thr) has an impact on development of hepatocellular carcinoma caused by chronic hepatitis C Egyptian patients. Subjects and Methods: The study included three groups; group 1: composing of 30 patients with hepatocellular carcinoma (HCC), group 2 composing of 30 patients with HCV, group 3 composing of 30 healthy subjects matching the same age and socioeconomic status were taken as a control group. Gene polymorphism of BCL2 (Ala43Thr) were evaluated by PCR-RFLP technique and measured for all patients and controls. Results: The summed 43Thr genotype was more frequent and statistically significant in HCC patients as compared to control group. This genotype of BCL2 gene may inhibit the programmed cell death which leads to disturbance in tissue and cells homeostasis and reduction in immune regulation. This result leads to viral replication and HCV persistence. Moreover, virus produces variety of mechanisms to block genes participated in apoptosis. This mechanism proves that HCV patients who have 43Thr genotype are more susceptible to HCC. Conclusion: The data suggest for the first time that the BCL2 polymorphism is associated with the susceptibility to HCC in Egyptian populations and might be used as molecular markers for evaluating HCC risk. This study clearly demonstrated that Chronic HCV exhibit a deregulation of apoptosis with the disease progression. This provides an insight into the pathogenesis of chronic HCV infection, and may contribute to the therapy.

Keywords: BCL2 gene, Hepatitis C Virus, Hepatocellular carcinoma, sensitivity, specificity, apoptosis

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284 Standardization of a Methodology for Quantification of Antimicrobials Used for the Treatment of Multi-Resistant Bacteria Using Two Types of Biosensors and Production of Anti-Antimicrobial Antibodies

Authors: Garzon V., Bustos R., Salvador J. P., Marco M. P., Pinacho D. G.

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Bacterial resistance to antimicrobial treatment has increased significantly in recent years, making it a public health problem. Large numbers of bacteria are resistant to all or nearly all known antimicrobials, creating the need for the development of new types of antimicrobials or the use of “last line” antimicrobial drug therapies for the treatment of multi-resistant bacteria. Some of the chemical groups of antimicrobials most used for the treatment of infections caused by multiresistant bacteria in the clinic are Glycopeptide (Vancomycin), Polymyxin (Colistin), Lipopeptide (Daptomycin) and Carbapenem (Meropenem). Molecules that require therapeutic drug monitoring (TDM). Due to the above, a methodology based on nanobiotechnology based on an optical and electrochemical biosensor is being developed, which allows the evaluation of the plasmatic levels of some antimicrobials such as glycopeptide, polymyxin, lipopeptide and carbapenem quickly, at a low cost, with a high specificity and sensitivity and that can be implemented in the future in public and private health hospitals. For this, the project was divided into five steps i) Design of specific anti-drug antibodies, produced in rabbits for each of the types of antimicrobials, evaluating the results by means of an immunoassay analysis (ELISA); ii) quantification by means of an electrochemical biosensor that allows quantification with high sensitivity and selectivity of the reference antimicrobials; iii) Comparison of antimicrobial quantification with an optical type biosensor; iv) Validation of the methodologies used with biosensor by means of an immunoassay. Finding as a result that it is possible to quantify antibiotics by means of the optical and electrochemical biosensor at concentrations on average of 1,000ng/mL, the antibodies being sensitive and specific for each of the antibiotic molecules, results that were compared with immunoassays and HPLC chromatography. Thus, contributing to the safe use of these drugs commonly used in clinical practice and new antimicrobial drugs.

Keywords: antibiotics, electrochemical biosensor, optical biosensor, therapeutic drug monitoring

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283 Synthesis and Characterization of Heterogeneous Silver Nanoparticles for Protection of Ancient Egyptian Artifacts from Microbial Deterioration

Authors: Mohamed Abd Elfattah Ibraheem Elghrbawy

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Biodeterioration of cultural heritage is a complex process which is caused by the interaction of many physical, chemical and biological agents; the growth of microorganisms can cause staining, cracking, powdering, disfigurement and displacement of monuments material, which leads to the permanent loss of monuments material. Organisms causing biodeterioration on monuments have usually been controlled by chemical products (biocides). In order to overcome the impact of biocides on the environment, human health and monument substrates, alternative tools such as antimicrobial agents from natural products can be used for monuments conservation and protection. The problem is how to formulate antibacterial agents with high efficiency and low toxicity. Various types of biodegradable metal nanoparticles (MNPs) have many applications in plant extract delivery. So, Nano-encapsulation of metal and natural antimicrobial agents using polymers such as chitosan increases their efficacy, specificity and targeting ability. Green synthesis and characterization of metal nanoparticles such as silver with natural products extracted from some plants having antimicrobial properties, using the ecofriendly method one pot synthesis. Encapsulation of the new synthesized mixture using some biopolymers such as chitosan nanoparticles. The dispersions and homogeneity of the antimicrobial heterogeneous metal nanoparticles encapsulated by biopolymers will be characterized and confirmed by Fourier Transform Infrared Spectroscopy (FTIR), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM) and Zeta seizer. The effect of the antimicrobial biopolymer metal nano-formulations on normal human cell lines will be investigated to evaluate the environmental safety of these formulations. The antimicrobial toxic activity of the biopolymeric antimicrobial metal nanoparticles formulations will be will be investigated to evaluate their efficiency towards different pathogenic bacteria and fungi.

Keywords: antimicrobial, biodeterioration, chitosan, cultural heritage, silver

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282 The Art of Resilience in the Case of Skopje

Authors: Kristina Nikolovska

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Social movements have become common in the Post Yugoslav cities. Consequently, the wave of activism has been considerably present in Skopje. Starting from 2009 the activist wave in Skopje emerged with the notion of the city. Diversity of initiatives appeared in the city in order to defend places that have been contested by the urban development project SK2014. The activist wave diffused into many different initiatives and diversity of issues. The result was unification in one massive movement in 2016, called 'The Colourful Revolution'. The paper explores the scope of activism in Skopje, with taking into consideration the influence of the spatial transformation, the project SK2014. Moreover, it examines the processes of spatiality into shaping the contention in Skopje, focusing on interdisciplinary and comprehensive approaches. Except the diversity of theoretical framework mainly founded on contentious politics theory and space elaboration from different perspectives, the study is founded on field work based on conducted interviews. Using an interdisciplinary approach and focusing on three main dimensions, the research contributes to understand the dynamics of the activist wave and importance of spatial processes in the creation of the contention in Skopje. Moreover, it elaborates the characteristics, possible effects, and reflections of the cycles of protests in Skopje. The main results of the research showed that dynamics of space is important in the creation of the activist wave in Skopje, moreover space context can give explanation about how opportunities diffuse and transformative power is created. The study contributed into deeper understanding of the importance of spatiality in contentious politics, it showed that in general contentions politics can benefit from deeper analyses of place specificity. Finally, the thesis opposes the traditional linear understanding of social movements, and proposes more dynamic, comprehensive, and sensitive elaboration.

Keywords: contentious politics, place, Skopje, SK2014, social movements, space

Procedia PDF Downloads 212
281 Maintenance Performance Measurement Derived Optimization: A Case Study

Authors: James M. Wakiru, Liliane Pintelon, Peter Muchiri, Stanley Mburu

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Maintenance performance measurement (MPM) represents an integrated aspect that considers both operational and maintenance related aspects while evaluating the effectiveness and efficiency of maintenance to ensure assets are working as they should. Three salient issues require to be addressed for an asset-intensive organization to employ an MPM-based framework to optimize maintenance. Firstly, the organization should establish important perfomance metric(s), in this case the maintenance objective(s), which they will be focuss on. The second issue entails aligning the maintenance objective(s) with maintenance optimization. This is achieved by deriving maintenance performance indicators that subsequently form an objective function for the optimization program. Lastly, the objective function is employed in an optimization program to derive maintenance decision support. In this study, we develop a framework that initially identifies the crucial maintenance performance measures, and employs them to derive maintenance decision support. The proposed framework is demonstrated in a case study of a geothermal drilling rig, where the objective function is evaluated utilizing a simulation-based model whose parameters are derived from empirical maintenance data. Availability, reliability and maintenance inventory are depicted as essential objectives requiring further attention. A simulation model is developed mimicking a drilling rig operations and maintenance where the sub-systems are modelled undergoing imperfect maintenance, corrective (CM) and preventive (PM), with the total cost as the primary performance measurement. Moreover, three maintenance spare inventory policies are considered; classical (retaining stocks for a contractual period), vendor-managed inventory with consignment stock and periodic monitoring order-to-stock (s, S) policy. Optimization results infer that the adoption of (s, S) inventory policy, increased PM interval and reduced reliance of CM actions offers improved availability and total costs reduction.

Keywords: maintenance, vendor-managed, decision support, performance, optimization

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280 Profile of Serological Response of Equids Naturally Infected with Burkholderia mallei

Authors: Iahtasham Khan, Vania Lucia De Assis Santana, Marcilia Maria Alves De Souza, Mabel Hanna Vance Harrop, Fernando Leandro Dos Santos, Cecília Maria Souza Leão E. Silva, Pedro Paulo Silveira, Marcelo Brasil, Marcus Vinícius, Hélio Cordeiro Manso Filho, Muhammad Younus, Aman Ullah Khan

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Glanders ranks high on clinical lists in some regions of Brazil as a cause of respiratory and lymphatic disease in equids. Glanders is caused by Burkolderia mallei (B. mallei) Gram-negative bacterium. B. mallei was first biological agent used in World War I in 20th century. The complement fixation test (CFT) is a serodiagnostic tool prescribed by the World Organization for Animal Health (OIE)for the diagnosis of glanders in the international trade of equids. The aim of the present study was to monitor the serological responses in equines naturally infected with B. mallei using the CFT. A total of 574 equids were tested with CFT, 30 days apart in a total of 12 samplings. One hundred thirty-four sera tested negative in all samplings; 192 sera tested positive in one sampling and 125 sera tested positive in two or more samplings. Remaining 123 samples showed uncertain results. Thus, CFT results can vary over a period of time. These variations could be the consequence of the effects of the natural immune response in each animal. The findings of the present study demonstrate difficulties regarding the simultaneous implementation of CFT and test and slaughter policies to eradicate glanders. Another constraint to control this disease is the presence of carrier/transitory CFT-negative animals, which are a potential source of disease in glanders-free areas. Serodiagnostic tests of higher sensitivity and specificity like immunobloat should be implemented to achieve success in the eradication of glanders.

Keywords: glanders, equids, horses, immunological, mules

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279 Shaping and Improving the Human Resource Management in Small and Medium Enterprises in Poland

Authors: Małgorzata Smolarek

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One of the barriers to the development of small and medium-sized enterprises (SME) are difficulties connected with management of human resources. The first part of article defines the specifics of staff management in small and medium enterprises. The practical part presents results of own studies in the area of diagnosis of the state of the human resources management in small and medium-sized enterprises in Poland. It takes into account its impact on the functioning of SME in a variable environment. This part presents findings of empirical studies, which enabled verification of the hypotheses and formulation of conclusions. The findings presented in this paper were obtained during the implementation of the project entitled 'Tendencies and challenges in strategic managing SME in Silesian Voivodeship.' The aim of the studies was to diagnose the state of strategic management and human resources management taking into account its impact on the functioning of small and medium enterprises operating in Silesian Voivodeship in Poland and to indicate improvement areas of the model under diagnosis. One of the specific objectives of the studies was to diagnose the state of the process of strategic management of human resources and to identify fundamental problems. In this area, the main hypothesis was formulated: The enterprises analysed do not have comprehensive strategies for management of human resources. The survey was conducted by questionnaire. Main Research Results: Human resource management in SMEs is characterized by simplicity of procedures, and the lack of sophisticated tools and its specificity depends on the size of the company. The process of human resources management in SME has to be adjusted to the structure of an organisation, result from its objectives, so that an organisation can fully implement its strategic plans and achieve success and competitive advantage on the market. A guarantee of success is an accurately developed policy of human resources management based on earlier analyses of the existing procedures and possessed human resources.

Keywords: human resources management, human resources policy, personnel strategy, small and medium enterprises

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278 Predicting Returns Volatilities and Correlations of Stock Indices Using Multivariate Conditional Autoregressive Range and Return Models

Authors: Shay Kee Tan, Kok Haur Ng, Jennifer So-Kuen Chan

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This paper extends the conditional autoregressive range (CARR) model to multivariate CARR (MCARR) model and further to the two-stage MCARR-return model to model and forecast volatilities, correlations and returns of multiple financial assets. The first stage model fits the scaled realised Parkinson volatility measures using individual series and their pairwise sums of indices to the MCARR model to obtain in-sample estimates and forecasts of volatilities for these individual and pairwise sum series. Then covariances are calculated to construct the fitted variance-covariance matrix of returns which are imputed into the stage-two return model to capture the heteroskedasticity of assets’ returns. We investigate different choices of mean functions to describe the volatility dynamics. Empirical applications are based on the Standard and Poor 500, Dow Jones Industrial Average and Dow Jones United States Financial Service Indices. Results show that the stage-one MCARR models using asymmetric mean functions give better in-sample model fits than those based on symmetric mean functions. They also provide better out-of-sample volatility forecasts than those using CARR models based on two robust loss functions with the scaled realised open-to-close volatility measure as the proxy for the unobserved true volatility. We also find that the stage-two return models with constant means and multivariate Student-t errors give better in-sample fits than the Baba, Engle, Kraft, and Kroner type of generalized autoregressive conditional heteroskedasticity (BEKK-GARCH) models. The estimates and forecasts of value-at-risk (VaR) and conditional VaR based on the best MCARR-return models for each asset are provided and tested using Kupiec test to confirm the accuracy of the VaR forecasts.

Keywords: range-based volatility, correlation, multivariate CARR-return model, value-at-risk, conditional value-at-risk

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277 Managing Company's Reputation during Crisis: An Analysis of Croatia Airlines' Crisis Response Strategy to the Labor Unions' Strike Announcement

Authors: M. Polic, N. Cesarec Salopek

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When it comes to crisis, no company, notwithstanding its financial success, power or reputation is immune to the new environment and circumstances emerging from it. The main challenge company faces with during a crisis is to protect its most valuable intangible asset reputation. Crisis has the serious potential to disrupt company’s everyday operations and damage its reputation extremely fast, especially if the company did not anticipate threats that may cause a crisis. Therefore, when a crisis happens, company must directly respond to it, whilst an effective crisis communication can limit consequences arising from the crisis, protect and repair the reputational damage caused to the company. Since every crisis is unique, each one of it requires different crisis response strategy. In July 2018, airline labor unions threatened Croatia Airlines, the state owned flag carrier of Croatia, to hold a strike that would be called into question regular flights and affect more than 7.600 passengers per day. This study explores the differences between crisis response strategies that Croatia Airlines, the state owned flag carrier of Croatia and airline labor unions used during the crisis period within the Situational Crisis Communication Theory (SCCT) by analyzing the content of formal communication tools used by Croatia Airlines and airline labor unions. Moreover, this study shows how Croatia Airlines successfully managed to communicate to the general public the threat that airline labor unions imposed on it and how was it received by the Croatian media. By using the qualitative and quantitative content analysis, the study will reveal the frames that dominated in the media articles during the crisis period. The greatest significance of this study is that it will provide the deeper insight into how transparent and consistent communication, the one that Croatia Airlines used before and during the crisis period, contributed to the decision of the competent court (Zagreb County Court) which prohibited labor unions strike in August 2018.

Keywords: crisis communication, crisis response strategy, Croatia Airlines, labor union, reputation management, situational crisis communication theory, strike

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276 Comparison of Formation Sensitivity Gap between Islamic Maybank Indonesia and Islamic Maybank Malaysia

Authors: Puji Sucia Sukmaningrum, Achsania Hendratmi, Noven Suprayogi, Muhammad Madyan

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Theoretically, Islamic banks in Indonesia and Malaysia not necessarily aware to the interest rate fluctuation, since they don’t use interest-based instruments. Both countries use dual banking system in which Islamic and conventional banking system are exist. This situation makes the profit-sharing level of the Islamic banks will be indirectly affected by the interest rate fluctuation from the conventional banks system. One of the risk management tools for anticipating the risk of interest rate fluctuation is gap management, which has purpose to narrow the difference between Rate Sensitive Asset (RSA) and Rate Sensitive Liability (RSL). This formed gap will give the information about the risk potential in Islamic banks which respect to the fluctuation on the interest rate. This study aims to determine the position of the gap formed at Islamic Maybank Indonesia and Islamic Maybank Malaysia, and analyze the difference in the formation of gap based on the period of sensitivity. This study is a quantitative research with comparative study using sensitivity gap analysis, independent sample t-test, and Mann-Whitney method. The data being used was secondary data from Maturity Profile contained in the Annual Financial Report of Islamic Maybank Indonesia and Islamic Maybank Malaysia from 2011 to 2015 period. The result shows that, cumulatively the formation of the gap was negative gap. From the results of independent sample t-test and Mann-Whitney, the formation of the gap in Islamic Maybank Indonesia and Islamic Maybank Malaysia for a period of sensitivity of ≤ 1 month and >1-3 months show a significant difference, while the period of sensitivity >3-12 months does not. The result shows, even though Indonesia and Malaysia using same dual banking systems, the gap values are different. The difference in debt policy between Indonesia and Malaysia also affecting the gap sensitivity in debt. In can be concluded that each country needs an appropriate gap management to support its Islamic banking performance specifically.

Keywords: assets and liability management, gap management, interest rate risk, Islamic bank

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275 Training for Digital Manufacturing: A Multilevel Teaching Model

Authors: Luís Rocha, Adam Gąska, Enrico Savio, Michael Marxer, Christoph Battaglia

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The changes observed in the last years in the field of manufacturing and production engineering, popularly known as "Fourth Industry Revolution", utilizes the achievements in the different areas of computer sciences, introducing new solutions at almost every stage of the production process, just to mention such concepts as mass customization, cloud computing, knowledge-based engineering, virtual reality, rapid prototyping, or virtual models of measuring systems. To effectively speed up the production process and make it more flexible, it is necessary to tighten the bonds connecting individual stages of the production process and to raise the awareness and knowledge of employees of individual sectors about the nature and specificity of work in other stages. It is important to discover and develop a suitable education method adapted to the specificities of each stage of the production process, becoming an extremely crucial issue to exploit the potential of the fourth industrial revolution properly. Because of it, the project “Train4Dim” (T4D) intends to develop complex training material for digital manufacturing, including content for design, manufacturing, and quality control, with a focus on coordinate metrology and portable measuring systems. In this paper, the authors present an approach to using an active learning methodology for digital manufacturing. T4D main objective is to develop a multi-degree (apprenticeship up to master’s degree studies) and educational approach that can be adapted to different teaching levels. It’s also described the process of creating the underneath methodology. The paper will share the steps to achieve the aims of the project (training model for digital manufacturing): 1) surveying the stakeholders, 2) Defining the learning aims, 3) producing all contents and curriculum, 4) training for tutors, and 5) Pilot courses test and improvements.

Keywords: learning, Industry 4.0, active learning, digital manufacturing

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274 Association Analysis of Putative Loci with Coronary Artery Disease

Authors: Asma Naseer Cheema, Attya Bhatti, Jabar Ali, John Peter

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Background: High cholesterol levels, endothelial dysfunction, inefficient coagulation cascade and hyper inflammatory response all are the basis of coronary artery disease (CAD). Several studies are carried out to see the genetic influence of these factors on disease outcome. Objective: The objective of our study was to see the association of 10 putative loci with coronary artery disease in our population. Materials & Methods: We screened our population for 10 putative loci of CAD showing significant association (p < 5x10-8) with candidate genes (regulating the cholesterol metabolism, endothelial function, coagulation cascade and inflammatory response of body). Hardy-Weinberg equilibrium and linkage disequilibrium in cases and controls s were estimated separately. Approximately 5-10 ng of dried DNA in 384 well plate format was used to genotype each sample on the Sequenom iPLEX assay at University of Pittsburgh Genomics and Proteomics Core Laboratories. It was built on single-base primer extension with the MALDI-TOF MS detection possessing high sensitivity and specificity. The SNPs were genotyped through Taqman assay. Hardy Weinberg test was applied. The 10 SNPs were selected as genetic markers for this study (rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650). Results: Mean age of the patient was 52 ± 11 years. Blood pressure and positive family history was found a significant risk factor for CAD. None of the selected SNPs showed significant association with coronary artery disease in our population (p>0.05). Conclusion: rs579459, rs1561198, rs2954029, rs1122608, rs17114036, rs9515203, rs10947789, rs7173743, rs2895811, rs2075650 are not significant genetic markers for CAD in our population.

Keywords: CAD, genetic markers, loci, risk factors

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273 Aminopeptidase P (DAP) Expression Pattern in Drosophila Melanogaster

Authors: Suneeta Gireesh Panicker

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Aim: Aminopeptidase P (APP) is an enzyme that has specificity for proline, can specifically cleave Xaa-Proline peptides and is a metallo-aminopeptidase. The bonds nearby to the imino acid proline are tough to cleave by many peptidases, but APP can specifically break peptide bonds engaged with proline. Membrane-bound form and a cytosolic form are the two forms in which this enzyme exists. The exact physiological function of APP remains unclear and hence the present work attempts to determine it. Methods: In the present study, the expression pattern of cytosolic Aminopeptidase P (DAP) was determined in all the embryonic stages and larval stages of wild-type Drosophila by using polyclonal monospecific antibodies. To show the presence of DAP RNA in embryonic and larval stages, RNA in situ hybridization was performed. DAP promoter-LacZ fusion reporter gene vector was used to construct transgenic embryos to study the regulation pattern of DAP. To study the DAP expression profile, a transgenic fly consisting of a DAP promoter with β-gal and GFP reporter genes in front of it was constructed. Results: DAP protein expression was observed in neuroectodermal cells, posterior midgut primordium, proctodeum, ventral neuroblast and primordial stomatogastric nervous system. It was observed in the ventral cord and midgut in stage 12. The completely developed embryos showed the intense occurrence of it in the ventral cord and gut region. The eye-antennal disc, wing disc and leg disc also showed the presence of DAP protein. LacZ expression in transgenic embryos also showed the same pattern. Conclusion: Similar to various known multiple-functional proteins, DAP could be one with different functions at different stages and in different cells. Data presented here designates DAP functions in the early embryonic and imaginal dics differentiation and development, suggesting that it may be required for the metabolism of proteins like neuropeptides and tachykinins.

Keywords: aminopeptidase P, in situ hybridization, transgenic fly, embryonic stages

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272 The Cleavage of DNA by the Anti-Tumor Drug Bleomycin at the Transcription Start Sites of Human Genes Using Genome-Wide Techniques

Authors: Vincent Murray

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The glycopeptide bleomycin is used in the treatment of testicular cancer, Hodgkin's lymphoma, and squamous cell carcinoma. Bleomycin damages and cleaves DNA in human cells, and this is considered to be the main mode of action for bleomycin's anti-tumor activity. In particular, double-strand breaks are thought to be the main mechanism for the cellular toxicity of bleomycin. Using Illumina next-generation DNA sequencing techniques, the genome-wide sequence specificity of bleomycin-induced double-strand breaks was determined in human cells. The degree of bleomycin cleavage was also assessed at the transcription start sites (TSSs) of actively transcribed genes and compared with non-transcribed genes. It was observed that bleomycin preferentially cleaved at the TSSs of actively transcribed human genes. There was a correlation between the degree of this enhanced cleavage at TSSs and the level of transcriptional activity. Bleomycin cleavage is also affected by chromatin structure and at TSSs, the peaks of bleomycin cleavage were approximately 200 bp apart. This indicated that bleomycin was able to detect phased nucleosomes at the TSSs of actively transcribed human genes. The genome-wide cleavage pattern of the bleomycin analogues 6′-deoxy-BLM Z and zorbamycin was also investigated in human cells. As found for bleomycin, these bleomycin analogues also preferentially cleaved at the TSSs of actively transcribed human genes. The cytotoxicity (IC₅₀ values) of these bleomycin analogues was determined. It was found that the degree of enhanced cleavage at TSSs was inversely correlated with the IC₅₀ values of the bleomycin analogues. This suggested that the level of cleavage at the TSSs of actively transcribed human genes was important for the cytotoxicity of bleomycin and analogues. Hence this study provided a deeper understanding of the cellular processes involved in the cancer chemotherapeutic activity of bleomycin.

Keywords: anti-tumour activity, bleomycin analogues, chromatin structure, genome-wide study, Illumina DNA sequencing

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271 Developing a Self-Healing Concrete Filler Using Poly(Methyl Methacrylate) Based Two-Part Adhesive

Authors: Shima Taheri, Simon Clark

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Concrete is an essential building material used in the majority of structures. Degradation of concrete over time increases the life-cycle cost of an asset with an estimated annual cost of billions of dollars to national economies. Most of the concrete failure occurs due to cracks, which propagate through a structure and cause weakening leading to failure. Stopping crack propagation is thus the key to protecting concrete structures from failure and is the best way to prevent inconveniences and catastrophes. Furthermore, the majority of cracks occur deep within the concrete in inaccessible areas and are invisible to normal inspection. Few materials intrinsically possess self-healing ability, but one that does is concrete. However, self-healing in concrete is limited to small dormant cracks in a moist environment and is difficult to control. In this project, we developed a method for self-healing of nascent fractures in concrete components through the automatic release of self-curing healing agents encapsulated in breakable nano- and micro-structures. The Poly(methyl methacrylate) (PMMA) based two-part adhesive is encapsulated in core-shell structures with brittle/weak inert shell, synthesized via miniemulsion/solvent evaporation polymerization. Stress fields associated with propagating cracks can break these capsules releasing the healing agents at the point where they are needed. The shell thickness is playing an important role in preserving the content until the final setting of concrete. The capsules can also be surface functionalized with carboxyl groups to overcome the homogenous mixing issues. Currently, this formulated self-healing system can replace up to 1% of cement in a concrete formulation. Increasing this amount to 5-7% in the concrete formulation without compromising compression strength and shrinkage properties, is still under investigation. This self-healing system will not only increase the durability of structures by stopping crack propagation but also allow the use of less cement in concrete construction, thereby adding to the global effort for CO2 emission reduction.

Keywords: self-healing concrete, concrete crack, concrete deterioration, durability

Procedia PDF Downloads 101