Search results for: target marketing

Authors: Hwang Ahreum, Kang Taejoon, Kim Bongsoo

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Nanosensors for high sensitive detection of diseases have been widely studied to improve the quality of life. Here, we suggest robust nano-plasmonic diagnostic sensor using cysteine tagged protein G (Cys3-protein G) and ultraflat, ultraclean and single-crystalline Au nanoplates. Protein G formed on an ultraflat Au surface provides ideal background for dense and uniform immobilization of antibodies. The Au is highly stable in diverse biochemical environment and can immobilize antibodies easily through Au-S bonding, having been widely used for various biosensing applications. Especially, atomically smooth single-crystalline Au nanomaterials synthesized using chemical vapor transport (CVT) method are very suitable to fabricate reproducible sensitive sensors. As the C-reactive protein (CRP) is a nonspecific biomarker of inflammation and infection, it can be used as a predictive or prognostic marker for various cardiovascular diseases. Cys3-protein G immobilized uniformly on the Au nanoplate enable CRP antibody (anti-CRP) to be ordered in a correct orientation, making their binding capacity be maximized for CRP detection. Immobilization condition for the Cys3-protein G and anti-CRP on the Au nanoplate is optimized visually by AFM analysis. Au nanoparticle - Au nanoplate (NPs-on-Au nanoplate) assembly fabricated from sandwich immunoassay for CRP can reduce zero-signal extremely caused by nonspecific bindings, providing a distinct surface-enhanced Raman scattering (SERS) enhancement still in 10-18 M of CRP concentration. Moreover, the NP-on-Au nanoplate sensor shows an excellent selectivity against non-target proteins with high concentration. In addition, comparing with control experiments employing a Au film fabricated by e-beam assisted deposition and linker molecule, we validate clearly contribution of the Au nanoplate for the attomolar sensitive detection of CRP. We expect that the devised platform employing the complex of single-crystalline Au nanoplates and Cys3-protein G can be applied for detection of many other cancer biomarkers.

Keywords: Au nanoplate, biomarker, diagnostic sensor, protein G, SERS

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Authors: Noor Abdelhamid, Donovan Nelson, Cara Prosser

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The architectural design process could be mapped out as a dialogue between designer and user that is constructed across multiple phases with the overarching goal of aligning design outcomes with user needs. Traditionally, this dialogue is bounded within a preliminary phase of determining factors that will direct the design intent, and a completion phase, of handing off the project to the client. Pre- and post-occupancy evaluations (P/POE’s) could provide an alternative process by extending this dialogue on both ends of the design process. The purpose of this research is to study the influence of systematic pre-occupancy data collection in achieving design goals by conducting post-occupancy evaluations of two case studies. In the context of this study, systematic pre-occupancy data collection is defined as the preliminary documentation of the existing conditions that helps portray stakeholders’ needs. When implemented, pre-occupancy occurs during the early phases of the architectural design process, utilizing the information to shape the design intent. Investigative POE’s are performed on two case studies with distinct early design approaches to understand how the current space is impacting user needs, establish design outcomes, and inform future strategies. The first case study underwent systematic pre-occupancy data collection and synthesis, while the other represents the traditional, uncoordinated practice of informally collecting data during an early design phase. POE’s target the dynamics between the building and its occupants by studying how spaces are serving the needs of the users. Data collection for this study consists of user surveys, audiovisual materials, and observations during regular site visits. Mixed methods of qualitative and quantitative analyses are synthesized to identify patterns in the data. The paper concludes by positioning value on both sides of the architectural design process: the integration of systematic pre-occupancy methods in the early phases and the reinforcement of a continued dialogue between building and design team after building completion.

Keywords: architecture, design process, pre-occupancy data, post-occupancy evaluation

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Authors: Tamunotonye Mayowa Braide, Isaac Oluwatayo

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This study investigates multidimensional poverty, and its correlates among rural households in Sekhukhune and Capricorn District municipalities (SDM & CDM) in Limpopo Province, South Africa. Primary data were collected from 407 rural households selected through purposive and simple random sampling techniques. Analytical techniques employed include descriptive statistics, principal component analysis (PCA), and the Alkire Foster (A-F) methodology. The results of the descriptive statistics showed there are more females (66%) than males (34%) in rural areas of Limpopo Province, with about 45% of them having secondary school education as the highest educational level attained and only about 3% do not have formal education. In the analysis of deprivation, eight dimensions of deprivation, constructed from 21 variables, were identified using the PCA. These dimensions include type and condition of dwelling water and sanitation, educational attainment and income, type of fuel for cooking and heating, access to clothing and cell phone, assets and fuel for light, health condition, crowding, and child health. In identifying the poor with poverty cut-off (0.13) of all indicators, about 75.9% of the rural households are deprived in 25% of the total dimensions, with the adjusted headcount ratio (M0) being 0.19. Multidimensional poverty estimates showed higher estimates of poor rural households with 71%, compared to 29%, which fall below the income poverty line. The study conducted poverty decomposition, using sub-groups within the area by examining regions and household characteristics. In SDM, there are more multidimensionally poor households than in CDM. The water and sanitation dimension is the largest contributor to the multidimensional poverty index (MPI) in rural areas of Limpopo Province. The findings can, therefore, assist in better design of welfare policy and target poverty alleviation programs and as well help in efficient resource allocation at the provincial and local municipality levels.

Keywords: Alkire-Foster methodology, Limpopo province, multidimensional poverty, principal component analysis, South Africa

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Authors: Mohit Wadhawan, Sushma Rathaur

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Lymphatic filariasis, also called elephantiasis is a tropical disease afflicting over 120 million people in 81 countries worldwide. Existing anti filarial drugs are effective against the larval stages of filarial parasites which call for an urgent need of drugs which are macrofilaricidal. Identification of molecular targets crucial for survival of filarial parasites is a prerequisite for drug designing. Prolyl oligopeptidase (POP) is one such crucial enzyme involved in the maturation and degradation of neuropeptides and peptide hormones. We have identified this peptidase in the bovine filarial parasite, Setaria cervi. Effect of inhibition of POP on the proteome profile of filarial parasite has been discussed in this study. Filarial parasites were exposed to Z-pro-prolinal (ZPP), a specific POP inhibitor for 8 h and the motility and viability of the parasites was observed. It significantly reduced the motility and viability of the parasites. To study the proteome profile, the cytosolic, endoplasmic reticulum (ER) and mitochondrial extracts of the adult female parasites were subjected to 2-dimensional electrophoresis. As analyzed by the PD-Quest software, the ZPP caused the alteration in the different subcellular proteins, and the significantly altered proteins were identified using MALDI-MS/MS spectrometry. The major proteins identified were found to play important role in diverse biological functions like signaling, redox regulation, energy metabolism, stress response, and cytoskeleton formation. Moreover, we found upregulation in the calcium binding proteins such as calreticulin, calponin, and calpain-6 suggesting that POP inhibition regulates calcium release. This relates to earlier reports that POP plays non-catalytic role in inositol 1,4,5-trisphosphate (IP3) signaling inducing release of calcium from ER. Taken together, the data demonstrated that inhibition of prolyl oligopeptidase alter the overall proteome signifying its role in survival of the filarial parasites. Thus this study provides a basis for the use of POP as a chemotherapeutic target for the treatment of lymphatic filariasis.

Keywords: lymphatic filariasis, setaria cervi, prolyl oligopeptidase, proteomics

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Authors: Farheen Tabassum, Shoab Ahmed Khan

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The brutality of attacks on networks and decisive infrastructures are on the climb over recent years and appears to continue to do so. Distributed Denial of service attack is the most prevalent and easy attack on the availability of a service due to the easy availability of large botnet computers at cheap price and the general lack of protection against these attacks. Application layer DDoS attack is DDoS attack that is targeted on wed server, application server or database server. These types of attacks are much more sophisticated and challenging as they get around most conventional network security devices because attack traffic often impersonate normal traffic and cannot be recognized by network layer anomalies. Conventional techniques of single-hosted security systems are becoming gradually less effective in the face of such complicated and synchronized multi-front attacks. In order to protect from such attacks and intrusion, corporation among all network devices is essential. To overcome this issue, a collaborative intrusion detection system (CIDS) is proposed in which multiple network devices share valuable information to identify attacks, as a single device might not be capable to sense any malevolent action on its own. So it helps us to take decision after analyzing the information collected from different sources. This novel attack detection technique helps to detect seemingly benign packets that target the availability of the critical infrastructure, and the proposed solution methodology shall enable the incident response teams to detect and react to DDoS attacks at the earliest stage to ensure that the uptime of the service remain unaffected. Experimental evaluation shows that the proposed collaborative detection approach is much more effective and efficient than the previous approaches.

Keywords: Distributed Denial-of-Service (DDoS), Collaborative Intrusion Detection System (CIDS), Slowloris, OSSIM (Open Source Security Information Management tool), OSSEC HIDS

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Authors: Sharmistha Chowdhury, Shuva Chowdhury

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Buying-selling new as well as second-hand goods like tools, furniture, household, electronics, clothing, baby stuff, vehicles, and hobbies through the Facebook marketplace has become a new paradigm for c2c sellers. This phenomenon encourages and empowers decentralised home-oriented sellers. This study adopts Elaboration Likelihood Model (ELM) to explain consumer behaviour on Facebook Marketplace (FM). ELM suggests that consumers process information through the central and peripheral routes, which eventually shape their attitudes towards posts. The central route focuses on information quality, and the peripheral route focuses on cues. Sellers’ FM posts usually include product features, prices, conditions, pictures, and pick-up location. This study uses information relevance and accuracy as central route factors. The post’s attractiveness represents cues and creates positive or negative associations with the product. A post with remarkable pictures increases the attractiveness of the post. So, post aesthetics is used as a peripheral route factor. People influenced via the central or peripheral route forms an attitude that includes multiple processes – response and purchase intention. People respond to FM posts through save, share and chat. Purchase intention reflects a positive image of the product and higher purchase intention. This study proposes trust on sellers as a moderator to test the strength of its influence on consumer attitudes and behaviour. Trust on sellers is assessed whether sellers have badges or not. A sample questionnaire will be developed and distributed among a group of random FM sellers who are selling vehicles on this platform to conduct the study. The chosen product of this study is the vehicle, a high-value purchase item. High-value purchase requires consumers to consider forming their attitude without any sign of impulsiveness seriously. Hence, vehicles are the perfect choice to test the strength of consumers attitudes and behaviour. The findings of the study add to the elaboration likelihood model and online second-hand marketplace literature.

Keywords: consumer behaviour, elaboration likelihood model, facebook marketplace, c2c marketing

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Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues

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Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.

Keywords: fish allergies, fish nutrition, proteomics, aquaculture

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Authors: Saraswati Suna

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The outbreak of the COVID-19 epidemic and the subsequent lockdown have significantly impacted India's political, structural, and economic systems and a rising gap between the rich and the disadvantaged, upper and lower caste. For Dalit women, such forms of subjugation were followed by socioeconomic uncertainty due to the pandemic's economic shutdown and labour oppressions. Dalit women have been the victims of the most oppression among the nation's underprivileged groups. Dalit women undergo systemic oppression at the hands of the state, caste, class, gender, and religious hegemons historically. Dalit women hold a subordinate position within the gender to their male counterparts and caste to their upper-caste counterparts. This paper examines how Brahminical patriarchy and state power severely affected Dalit/Adivasi women during COVID-19 in Odisha, India. In order to understand caste-based sexual violence, a total of five cases have been analysed from newspapers. Findings revealed that Covid-19 appears to have a significant physical, psychological, and economic impact on Dalit women. The intention of sexual harassment and rape perpetrated by upper caste men is to maintain power and patriarchal culture in society. Dalit women are economically, socially, and culturally marginalised, which effectively exacerbates the sense of impunity by perpetrators of violence against Dalit women. This issue requires special attention to end atrocities against Dalit women. Dalit women become the target of rape, sexual assault, and murder. Sexual violence against Dalit women cannot be fully explained without linkage to caste, gender, and power. Dominant caste comes through caste privilege-socio-economic and politically; these factors contribute to sexual violence against Dalit women. The findings revealed that state police manipulate sexual violence, and in so doing, they create and deny access to both services to get justice. This article has argued that understanding Brahminical culture and the legal impacts of state police on Dalit women's identity requires a nuanced analysis.

Keywords: COVID-19, dalit women, sexual violence, brahminical patriarchy, power

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Authors: Małgorzata Tomczyńska, Joanna Saluk-Bijak

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Background: Graves’ disease is a frequent organ-specific autoimmune thyroid disease, which characterized by the presence of different kind autoantibodies, that, in most cases, act as agonists of the thyrotropin receptor, leading to hyperthyroidism. Role of platelets and lymphocytes can be modulated in the pathophysiology of thyroid autoimmune diseases. Interference in the physiology of platelets can lead to enhanced activity of these cells. Activated platelets can bind to circulating lymphocytes and to affect lymphocyte adhesion. Platelets and lymphocytes can regulate mutual functions. Therefore, the activation of T lymphocytes, as well as blood platelets, is associated with the development of inflammation and oxidative stress within the target tissue. The present study was performed to investigate a platelet-lymphocyte relation by assessing the degree of their mutual aggregation in whole blood of patients with Graves’ disease. Also, the purpose of this study was to examine the impact of platelet interaction on lymphocyte migration capacity. Methods: 30 patients with Graves’ disease were recruited in the study. The matched 30 healthy subjects were served as the control group. Immunophenotyping of lymphocytes was carried out by flow cytometry method. A CytoSelect™ Cell Migration Assay Kit was used to evaluate lymphocyte migration and adhesion to blood platelets. Visual assessment of lymphocyte-platelet aggregate morphology was done using confocal microscope after magnetic cell isolation by Miltenyi Biotec. Results: The migration and functional responses of lymphocytes to blood platelets were greater in the group of Graves’ disease patients compared with healthy controls. The group of Graves’ disease patients exhibited a reduced T lymphocyte and a higher B cell count compared with controls. Based on microscopic analysis, more platelet-lymphocyte aggregates were found in patients than in control. Conclusions: Studies have shown that in Graves' disease, lymphocytes show increased platelet affinity, more strongly migrating toward them, and forming mutual cellular conglomerates. This may be due to the increased activation of blood platelets in this disease.

Keywords: blood platelets, cell migration, Graves’ disease, lymphocytes, lymphocyte-platelet aggregates

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Authors: Alexandra P. Alexandropoulou, Andreas Fousteris, Eleni Didaskalou, Dimitrios A. Georgakellos

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As urbanization is becoming a major stress factor not only for the urban environment but also for the wellbeing of city dwellers, incorporating the issues of urban health in strategic city planning and policy-making has never been more relevant. The impact of urbanization can vary from low to severe and relates to all non-communicable diseases caused by the different functions of cities. Air pollution, noise pollution, water and soil pollution, availability of open green spaces, and urban heat island are the major factors that can compromise citizens' health. Urban health describes the effects of the social environment, the physical environment, and the availability and accessibility to health and social services. To assess the quality of urban wellbeing, all urban characteristics that might have an effect on citizens' health must be considered, evaluated, and introduced in integrated local planning. A series of indices and indicators can be used to better describe these effects and set the target values in policy making. Local strategic planning is one of the most valuable development tools a local city administration can possess; thus, it has become mandatory under Greek law for all municipalities. It involves a two-stage procedure; the first aims to collect, analyse and evaluate data on the current situation of the city (administrative data, population data, environmental data, social data, swot analysis), while the second aims to introduce a policy vision described and supported by distinct (nevertheless integrated) actions, plans and measures to be implemented with the aim of city development and citizen wellbeing. In this procedure, the element of health is often neglected or under-evaluated. A relative survey was conducted among all Greek local authorities in order to shed light on the current situation. Evidence shows that the rate of incorporation of health in strategic planning is lacking behind. The survey also highlights key hindrances and concerns raised by local officials and suggests a path for the way forward.

Keywords: urban health, strategic planning, local authorities, integrated development

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Authors: Armando A. Rodríguez, Emilio Salceda, Anoland Garateix, André J. Zaharenko, Steve Peigneur, Omar López, Tirso Pons, Michael Richardson, Maylín Díaz, Yasnay Hernández, Ludger Ständker, Jan Tytgat, Enrique Soto

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Acid-sensing ion channels are cation (Na+) channels activated by a pH drop. These proteins belong to the ENaC/degenerin superfamily of sodium channels. ASICs are involved in sensory perception, synaptic plasticity, learning, memory formation, cell migration and proliferation, nociception, and neurodegenerative disorders, among other processes; therefore those molecules that specifically target these channels are of growing pharmacological and biomedical interest. Sea anemones produce a large variety of ion channels peptide toxins; however, those acting on ligand-gated ion channels, such as Glu-gated, Ach-gated ion channels, and acid-sensing ion channels (ASICs), remain barely explored. The peptide PhcrTx1 is the first compound characterized from the sea anemone Phymanthus crucifer, and it constitutes a novel ASIC inhibitor. This peptide was purified by chromatographic techniques and pharmacologically characterized on acid-sensing ion channels of mammalian neurons using patch-clamp techniques. PhcrTx1 inhibited ASIC currents with an IC50 of 100 nM. Edman degradation yielded a sequence of 32 amino acids residues, with a molecular mass of 3477 Da by MALDI-TOF. No similarity to known sea anemone peptides was found in protein databases. The computational analysis of Cys-pattern and secondary structure arrangement suggested that this is a structurally ICK (Inhibitor Cystine Knot)-type peptide, a scaffold that had not been found in sea anemones but in other venomous organisms. These results show that PhcrTx1 represents the first member of a new structural group of sea anemones toxins acting on ASICs. Also, this peptide constitutes a novel template for the development of drugs against pathologies related to ASICs function.

Keywords: animal toxin, inhibitor cystine knot, ion channel, sea anemone

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Authors: Gezahegn Gizaw

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The FDRE Tourism Training Institute organizes forums for debates, best practices exchange and focus group discussions to forge a sustainable and growing tourism sector while minimizing negative impacts on the environment, communities, and cultures. This study aimed at applying empirical research method to identify and quantify relative importance of success factors and individual engagement indicators that were identified in these forums. Response to the 12-question survey was collected from a total of 437 respondents in academic training institutes (212), business executive and employee (204) and non-academic government offices (21). Overall, capacity building was perceived as the most important driver of success for stakeholder engagement. Business executive and employee category rated capacity building as the most important driver of success (53%), followed by decision-making process (27%) and community participation (20%). Among educators and students, both capacity building and decision-making process were perceived as the most important factors (40% of respondents), whereas community participation was perceived as the most important success factor only by 20% of respondents. Individual engagement score in capacity building, decision-making process and community participation showed highest variability by educational level of participants (variance of 3.4% - 5.2%, p<0.001). Individual engagement score in capacity building was highly correlated to perceived benefit of training on improved efficiency, job security, higher customer satisfaction and self-esteem. On the other hand, individual engagement score in decision making process was highly correlated to its perceived benefit on lowering business costs, improving ability to meet the needs of a target market, job security, self-esteem and more teamwork. The study provides a set of recommendations that help educators, business executives and policy makers to maximize the individual and synergetic effect of training, decision making process on sustainability and growth of the tourism sector in Ethiopia.

Keywords: engagement score, driver of success, capacity building, tourism

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Authors: Merfat Algethami, Anton Blencowe, Bryce Feltis, Stephen Best, Moshi Geso

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Nano-materials with high atomic number atoms have been demonstrated to enhance the effective radiation dose and thus potentially could improve therapeutic efficacy in radiotherapy. The optimal nanoparticulate agents require high X-ray absorption coefficients, low toxicity, and should be cost effective. The focus of our research is the development of a nanoparticle therapeutic agent that can be used in radiotherapy to provide optimal enhancement of the radiation effects on the target. In this study, we used bismuth (Bi) nanoparticles coated with starch and bismuth sulphide nanoparticles (Bi2S3) coated with polyvinylpyrrolidone (PVP). These NPs are of low toxicity and are one of the least expensive heavy metal-based nanoparticles. The aims of this study were to synthesise Bi2S3 and Bi NPs, and examine their cytotoxicity to human lung adenocarcinoma epithelial cells (A549). The dose enhancing effects of NPs on A549 cells were examined at both KV and MV energies. The preliminary results revealed that bismuth based nanoparticles show increased radio-sensitisation of cells, displaying dose enhancement with KV X-ray energies and to a lesser degree for the MV energies. We also observed that Bi NPs generated a greater dose enhancement effect than Bi2S3 NPs in irradiated A549 cells. The maximum Dose Enhancement Factor (DEF) was obtained at lower energy KV range when cells treated with Bi NPs (1.5) compared to the DEF of 1.2 when cells treated with Bi2S3NPs. Less radiation dose enhancement was observed when using high energy MV beam with higher DEF value of Bi NPs treatment (1.26) as compared to 1.06 DEF value with Bi2S3 NPs. The greater dose enhancement was achieved at KV energy range, due the effect of the photoelectric effect which is the dominant process of interaction of X-ray. The cytotoxic effect of Bi NPs on enhancing the X-ray dose was higher due to the higher amount of elemental Bismuth present in Bi NPs compared to Bi2S3 NPs. The results suggest that Bismuth based NPs can be considered as valuable dose enhancing agents when used in clinical applications.

Keywords: A549 lung cancer cells, Bi2S3 nanoparticles, dose enhancement effect, radio-sensitising agents

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Authors: Abbas Ali Husseini, Ali Mohammad Yazdani, Fatemeh Ghadiri, Alper Şişman

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We developed a microchip device that uses surface acoustic waves for rapid lysis of low level of cell samples. The device incorporates sharp-edge glass microparticles for improved performance. We optimized the lysis conditions for high efficiency and evaluated the device's feasibility for point-of-care applications. The microchip contains a 13-finger pair interdigital transducer with a 30-degree focused angle. It generates high-intensity acoustic beams that converge 6 mm away. The microchip operates at a frequency of 16 MHz, exciting Rayleigh waves with a 250 µm wavelength on the LiNbO3 substrate. Cell lysis occurs when Candida albicans cells and glass particles are placed within the focal area. The high-intensity surface acoustic waves induce centrifugal forces on the cells and glass particles, resulting in cell lysis through lateral forces from the sharp-edge glass particles. We conducted 42 pilot cell lysis experiments to optimize the surface acoustic wave-induced streaming. We varied electrical power, droplet volume, glass particle size, concentration, and lysis time. A regression machine-learning model determined the impact of each parameter on lysis efficiency. Based on these findings, we predicted optimal conditions: electrical signal of 2.5 W, sample volume of 20 µl, glass particle size below 10 µm, concentration of 0.2 µg, and a 5-minute lysis period. Downstream analysis successfully amplified a DNA target fragment directly from the lysate. The study presents an efficient microchip-based cell lysis method employing acoustic streaming and microparticle collisions within microdroplets. Integration of a surface acoustic wave-based lysis chip with an isothermal amplification method enables swift point-of-care applications.

Keywords: cell lysis, surface acoustic wave, micro-glass particle, droplet

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Authors: David W. Taylor, Fernando Lourenço, Carolyn Branston, Paul Tucker

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There are an increasing number of students at Universities in the United Kingdom engaging in entrepreneurship role-play to explore business start-up as a career alternative to employment. These role-play activities have been shown to have a positive influence on students’ entrepreneurial intentions. Universities also play a role in developing graduates’ awareness of social responsibility. However, social responsibility is often missing from these entrepreneurship role-plays. It is important that these role-play activities include the development of values that support social responsibility, in-line with those running hybrid, humane and sustainable enterprises, and not simply focus on profit. The Young Enterprise (YE) Start-Up programme is an example of a role-play activity that is gaining in popularity amongst United Kingdom Universities seeking ways to give students insight into a business start-up. A Post-92 University in the North-West of England has adapted the traditional YE Directorship roles (e.g., Marketing Director, Sales Director) by including a Corporate Social Responsibility (CSR) Director in all of the team-based YE Start-Up businesses. The aim for introducing this Directorship was to observe if such a role would help create a more socially responsible value-system within each company and in turn shape business decisions. This paper investigates role-play as a tool to help enterprise educators develop socially responsible attitudes and values in nascent entrepreneurs. A mixed qualitative methodology approach has been used, which includes interviews, role-play, and reflection, to help students develop positive value characteristics through the exploration of unethical and selfish behaviors. The initial findings indicate that role-play helped CSR Directors learn and gain insights into the importance of corporate social responsibility, influenced the values and actions of their YE Start-Ups, and increased the likelihood that if the participants were to launch a business post-graduation, that the intent would be for the business to be socially responsible. These findings help inform educators on how to develop socially responsible nascent entrepreneurs within a traditionally profit orientated business model.

Keywords: student entrepreneurship, young enterprise, social responsibility, role-play, values

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Authors: Davod Ahmadi, Narges Ebadi, Ethan Wang, Hugo Melgar-Quiñonez

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Compared to the other Least Developed Countries (LDCs), major countries in sub-Saharan Africa (SSA) have limited access to the clean water. People in this region, and more specifically females, suffer from acute water scarcity problems. They are compelled to spend too much of their time bringing water for domestic use like drinking and washing. Apart from domestic use, water through affecting agriculture and livestock contributes to the food security status of people in vulnerable regions like SSA. Livestock needs water to grow, and agriculture requires enormous quantities of water for irrigation. The main objective of this study is to explore the association between access to water and individuals’ food security status. Data from 2017 Gallup World Poll (GWP) for SSA were analyzed (n=35,000). The target population in GWP is the entire civilian, non-institutionalized, aged 15 and older population. All samples selection is probability based and nationally representative. The Gallup surveys an average of 1,000 samples of individuals per country. Three questions related to water (i.e., water quality, availability of water for crops and availability of water for livestock) were used as the exposure variables. Food Insecurity Experience Scale (FIES) was used as the outcome variable. FIES measures individuals’ food security status, and it is composed of eight questions with simple dichotomous responses (1=Yes and 0=No). Different statistical analyses such as descriptive, crosstabs and binary logistic regression, form the basis of this study. Results from descriptive analyses showed that more than 50% of the respondents had no access to enough water for crops and livestock. More than 85% of respondents were categorized as “food insecure”. Findings from cross-tabulation analyses showed that food security status was significantly associated with water quality (0.135; P=0.000), water for crops (0.106; P=0.000) and water for livestock (0.112; P=0.000). In regression analyses, the probability of being food insecure increased among people who expressed no satisfaction with water quality (OR=1.884 (OR=1.768-2.008)), not enough water for crops (OR=1.721 (1.616-1.834)) and not enough water for livestock (OR=1.706 (1.819)). In conclusion, it should note that water access affects food security status in SSA.

Keywords: water access, agriculture, livestock, FIES

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Authors: R. Fardid, R. Coppes

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Introduction: In mediastinal radiotherapy and to a lesser extend also in total-body irradiation (TBI) radiation exposure may lead to development of cardiac diseases. Radiation-induced heart disease is dose-dependent and it is characterized by a loss of cardiac function, associated with progressive heart cells degeneration. We aimed to determine the in-vivo radiation effects on fibronectin, ColaA1, ColaA2, galectin and TGFb1 gene expression levels in left ventricle heart tissues of rats after irradiation. Material and method: Four non-treatment adult Wistar rats as control group (group A) were selected. In group B, 4 adult Wistar rats irradiated to 20 Gy single dose of 150 Mev proton beam locally in heart only. In heart plus lung irradiate group (group C) 4 adult rats was irradiated by 50% of lung laterally plus heart radiation that mentioned in before group. At 8 weeks after radiation animals sacrificed and left ventricle heart dropped in liquid nitrogen for RNA extraction by Absolutely RNA® Miniprep Kit (Stratagen, Cat no. 400800). cDNA was synthesized using M-MLV reverse transcriptase (Life Technologies, Cat no. 28025-013). We used Bio-Rad machine (Bio Rad iQ5 Real Time PCR) for QPCR testing by relative standard curve method. Results: We found that gene expression of fibronectin in group C significantly increased compared to control group, but it was not showed significant change in group B compared to group A. The levels of gene expressions of Cola1 and Cola2 in mRNA did not show any significant changes between normal and radiation groups. Changes of expression of galectin target significantly increased only in group C compared to group A. TGFb1 expressions in group C more than group B showed significant enhancement compared to group A. Conclusion: In summary we can say that 20 Gy of proton exposure of heart tissue may lead to detectable damages in heart cells and may distribute function of them as a component of heart tissue structure in molecular level.

Keywords: gene expression, heart damage, proton irradiation, radiotherapy

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Authors: Zahid Ullah, Atlas Khan

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The rapid growth of data in diverse domains has created an urgent need for effective utilization of mathematical and statistical sciences in data analytics. This abstract explores the journey from theory to practice, emphasizing the importance of harnessing mathematical and statistical innovations to unlock the full potential of data analytics. Drawing on a comprehensive review of existing literature and research, this study investigates the fundamental theories and principles underpinning mathematical and statistical sciences in the context of data analytics. It delves into key mathematical concepts such as optimization, probability theory, statistical modeling, and machine learning algorithms, highlighting their significance in analyzing and extracting insights from complex datasets. Moreover, this abstract sheds light on the practical applications of mathematical and statistical sciences in real-world data analytics scenarios. Through case studies and examples, it showcases how mathematical and statistical innovations are being applied to tackle challenges in various fields such as finance, healthcare, marketing, and social sciences. These applications demonstrate the transformative power of mathematical and statistical sciences in data-driven decision-making. The abstract also emphasizes the importance of interdisciplinary collaboration, as it recognizes the synergy between mathematical and statistical sciences and other domains such as computer science, information technology, and domain-specific knowledge. Collaborative efforts enable the development of innovative methodologies and tools that bridge the gap between theory and practice, ultimately enhancing the effectiveness of data analytics. Furthermore, ethical considerations surrounding data analytics, including privacy, bias, and fairness, are addressed within the abstract. It underscores the need for responsible and transparent practices in data analytics, and highlights the role of mathematical and statistical sciences in ensuring ethical data handling and analysis. In conclusion, this abstract highlights the journey from theory to practice in harnessing mathematical and statistical sciences in data analytics. It showcases the practical applications of these sciences, the importance of interdisciplinary collaboration, and the need for ethical considerations. By bridging the gap between theory and practice, mathematical and statistical sciences contribute to unlocking the full potential of data analytics, empowering organizations and decision-makers with valuable insights for informed decision-making.

Keywords: data analytics, mathematical sciences, optimization, machine learning, interdisciplinary collaboration, practical applications

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Authors: Umar Saeed

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Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

Procedia PDF Downloads 55

Authors: Fei Sun, Meilan Xu, Jianghui Zhu, Maria Stefanie Dwiyanti, Cheolwoo Park, Fanjiang Kong, Baohui Liu, Tetsuya Yamada, Jun Abe

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Reduced or lack of sensitivity to long daylengths is a key character for soybean, a short-day crop, to adapt to higher latitudinal environments. However, the photoperiod-insensitivity often results in a reduction of the duration of vegetative growth and final yield. To overcome this limitation, a photoperiod insensitive line (RIL16) was developed in this study that delayed flowering from the recombinant inbred population derived from a cross between a photoperiod-insensitive cultivar AGS292 and a late-flowering Thai cultivar K3. Expression analyses under SD and LD conditions revealed that the expression levels of FLOWERING LOCUS T (FT) orthologues, FT2a and FT5a, were lowered in RIL16 relative to AGS292, although the expression of E1, a soybean-specific suppressor for FTs, was inhibited in both conditions. A soybean orthologue of TARGET OF EAT1 (TOE1), another suppressor of FT, showed an upregulated expression in RIL16, which appeared to reflect a lower expression of miR172a. Our data suggest that the delayed flowering of RIL16 most likely is controlled by genes involved in an age-dependent pathway in flowering. The QTL analysis based on 1,125 SNPs obtained from Restriction Site Associated DNA Sequencing revealed two major QTLs for flowering dates in Chromosome 16 and two minor QTLs in Chromosome 4, all of which accounted for 55% and 48% of the whole variations observed in natural day length and artificially-induced long day length conditions, respectively. The intervals of the major QTLs harbored FT2a and FT5a, respectively, on the basis of annotated genes in the Williams 82 reference genome. Sequencing analysis further revealed a nonsynonymous mutation in FT2a and an SNP in the 3′ UTR region of FT5a. A further study may elucidate a detailed mechanism underlying the QTL for late flowering. The alleles from K3 at the two QTLs can be used singly or in combination to retain an appropriate duration of vegetative growth to maximize the final yield of photoperiod-insensitive soybeans.

Keywords: FT genes, miR72a, photoperiod-insensitive, soybean flowering

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Authors: Xie Xiaobin, Zhang Yan, Shi Yipeng, Sun Wenying, Chen Shuang, Cai Zhipeng, Zhang Hong, Zhang Lixia, Xie Like

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Objective: The objective of this research is to compare the efficacy of the late in-situ surgical revision augmented with continuous infusion and needle revision on failed trabeculectomy blebs. Methods From December 2018 to December 2021, a prospective randomized controlled trial was performed on 44 glaucoma patients with failed bleb ≥ 6months with medically uncontrolled in Eye Hospital, China Academy of Chinese Medical Sciences. They were randomly divided into two groups. 22 eyes of 22 patients underwent the late in-situ surgical revision with continuous anterior chamber infusion in the study group, and 22 of 22 patients were treated with needle revision in the control group. Main outcome measures include preoperative and postoperative intraocular pressure (IOP), the number of anti-glaucoma medicines, the operation success rate, and the postoperative complications. Results The postoperative IOP values decreased significantly from the baseline in both groups (both P<0.05). IOP was significantly lower in the study group than in the control group at one week, 1, and 3 months postoperatively (all P<0.05). IOP reductions in the study group were substantially more prominent than in the control group at all postoperative time points (all P<0.05). The complete success rate in the study group was significantly higher than in the control group (71.4% vs. 33.3%, P<0.05), while the complete failure rate was significantly lower in the study group (0% vs. 28.5%, P<0.05). According to Cox’s proportional hazards regression analysis, high IOP at baseline was independently associated with increased risks of complete failure (adjusted hazard ratio=1.141, 95% confidence interval=1.021-1.276, P<0.05). There was no significant difference in the incidence of postoperative complications between the two groups (P>0.05). Conclusion: Both in-situ surgical and needle revision have acceptable success rates and safety for the late failed trabeculectomy blebs, while the former is likely to have a higher level of efficacy over the latter. Needle revision may be insufficient for eyes with low target IOP.

Keywords: glaucoma, trabeculectomy blebs, in-situ surgical revision, needle revision

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Authors: Yasmin Begum Mohammed

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Ketorolac tromethamine (KTM) is a non-steroidal anti-inflammatory drug with a potent analgesic and anti-inflammatory activity due to prostaglandin related inhibitory effect of drug. It is a non-selective cyclo-oxygenase inhibitor. The drug is currently used orally and intramuscularly in multiple divided doses, clinically for the management arthritis, cancer pain, post-surgical pain, and in the treatment of migraine pain. KTM has short biological half-life of 4 to 6 hours, which necessitates frequent dosing to retain the action. The frequent occurrence of gastrointestinal bleeding, perforation, peptic ulceration, and renal failure lead to the development of other drug delivery strategies for the appropriate delivery of KTM. The ideal solution would be to target the drug only to the cells or tissues affected by the disease. Drug targeting could be achieved effectively by liposomes that are biocompatible and biodegradable. The aim of the study was to develop a parenteral liposome formulation of KTM with improved efficacy while reducing side effects by targeting the inflammation due to arthritis. PEG-anchored (stealth) and non-PEG-anchored liposomes were prepared by thin film hydration technique followed by extrusion cycle and characterized for in vitro and in vivo. Stealth liposomes (SLs) exhibited increase in percent encapsulation efficiency (94%) and 52% percent of drug retention during release studies in 24 h with good stability for a period of 1 month at -20°C and 4°C. SLs showed about maximum 55% of edema inhibition with significant analgesic effect. SLs produced marked differences over those of non-SL formulations with an increase in area under plasma concentration time curve, t₁/₂, mean residence time, and reduced clearance. 0.3% of the drug was detected in arthritic induced paw with significantly reduced drug localization in liver, spleen, and kidney for SLs when compared to other conventional liposomes. Thus SLs help to increase the therapeutic efficacy of KTM by increasing the targeting potential at the inflammatory region.

Keywords: biodistribution, ketorolac tromethamine, stealth liposomes, thin film hydration technique

Procedia PDF Downloads 282

Authors: Gift Chidi-Onwuta, Iwe Nkem Nkechinyere, Chikamadu Christabelle Chinyere

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The teaching of English is a resource-driven activity that requires rich resource-classroom settings for the delivery of effective lessons and the acquisition of interpersonal skills for integration in a target-language environment. However, throughout the world, English is often taught in low-resource classrooms. This paper is aimed to reveal the common problems associated with teaching English in low-resource environments and the prospects for teachers who found themselves in such undefined teaching settings. Self-structured and validated questionnaire in a closed-ended format, open question format and scaling format was administered to teachers across five countries: Nigeria, Cameroun, Iraq, Turkey, and Sudan. The study adopts situational language teaching theory (SLTT), which emphasizes a performance improvement imperative. This study inclines to this model because it maintains that learning must be fun and enjoyable like playing a favorite sport, just as in real life. Since teaching resources make learning engaging, we found this model apt for the current study. The perceptions of teachers about accessibility and functionality of teaching material resources, the nature of teaching outcomes in resource-less environments, their levels of involvement in improvisation and the prospects associated with resource limitations were sourced. Data were analysed using percentages and presented in frequency tables. Results: showed that a greater number of teachers across these nations do not have access to sufficient productive resource materials that can aid effective English language teaching. Teaching outcomes, from the findings, are affected by low material resources; however, results show certain advantages to teaching English with limited resources: flexibility and autonomy with students and creativity and innovation amongst teachers. Results further revealed group work, story, critical thinking strategy, flex, cardboards and flashcards, dictation and dramatization as common teaching strategies, as well as materials adopted by teachers to overcome low resource-related challenges in classrooms.

Keywords: teaching materials, low-resource environments, English language teaching, situational language theory

Procedia PDF Downloads 117

Authors: Y. A. Prada, Fanny Guzman, Rafael Cabanzo, John J. Castillo, Enrique Mejia-Ospino

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In this work, we show the important results about of synthesis of photoluminiscents gold nanoclusters using a small peptide as template for biosensing applications. Interestingly, we design one peptide (NBC2854) homologue to conservative domain from 215 250 residue of a galactolectin protein which can recognize the proteophosphoglycans (PPG) from Leishmania. Peptide was synthetized by multiple solid phase synthesis using FMoc group methodology in acid medium. Finally, the peptide was purified by High-Performance Liquid Chromatography using a Vydac C-18 preparative column and the detection was at 215 nm using a Photo Diode Array detector. Molecular mass of this peptide was confirmed by MALDI-TOF and to verify the α-helix structure we use Circular Dichroism. By means of the methodology used we obtained a novel fluorescents gold nanoclusters (AuNC) using NBC2854 as a template. In this work, we described an easy and fast microsonic method for the synthesis of AuNC with ≈ 3.0 nm of hydrodynamic size and photoemission at 630 nm. The presence of cysteine residue in the C-terminal of the peptide allows the formation of Au-S bond which confers stability to Peptide-based gold nanoclusters. Interactions between the peptide and gold nanoclusters were confirmed by X-ray Photoemission and Raman Spectroscopy. Notably, from the ultrafine spectra shown in the MALDI-TOF analysis which containing only 3-7 KDa species was assigned to Au₈-₁₈[NBC2854]₂ clusters. Finally, we evaluated the Peptide-gold nanocluster as an optical biosensor based on fluorescence spectroscopy and the fluorescence signal of PPG (0.1 µg-mL⁻¹ to 1000 µg-mL⁻¹) was amplified at the same wavelength emission (≈ 630 nm). This can suggest that there is a strong interaction between PPG and Pep@AuNC, therefore, the increase of the fluorescence intensity can be related to the association mechanism that take place when the target molecule is sensing by the Pep@AuNC conjugate. Further spectroscopic studies are necessary to evaluate the fluorescence mechanism involve in the sensing of the PPG by the Pep@AuNC. To our best knowledge the fabrication of an optical biosensor based on Pep@AuNC for sensing biomolecules such as Proteophosphoglycans which are secreted in abundance by parasites Leishmania.

Keywords: biosensing, fluorescence, Leishmania, peptide-gold nanoclusters, proteophosphoglycans

Procedia PDF Downloads 156

Authors: M. F. Slim, A. Alhussein, F. Sanchette, M. François

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Thin films are widely used in various applications to enhance the surface properties and characteristics of materials. They are used in many domains such as: biomedical, automotive, aeronautics, military, electronics and energy. Depending on the elaboration technique, the elastic behavior of thin films may be different from this of bulk materials. This dependence on the elaboration techniques and their parameters makes the control of the elasticity constants of coated components necessary. Our work is focused on the characterization of the elasticity constants of isotropic thin films by means of Impulse Excitation Techniques. The tests rely on the measurement of the sample resonance frequency before and after deposition. In this work, a finite element model was performed with ABAQUS software. This model was then compared with the analytical approaches used to determine the Young’s and shear moduli. The best model to determine the film Young’s modulus was identified and a relation allowing the determination of the shear modulus of thin films of any thickness was developed. In order to confirm the model experimentally, Tungsten films were deposited on glass substrates by DC magnetron sputtering of a 99.99% purity tungsten target. The choice of tungsten was done because it is well known that its elastic behavior at crystal scale is ideally isotropic. The macroscopic elasticity constants, Young’s and shear moduli and Poisson’s ratio of the deposited film were determined by means of Impulse Excitation Technique. The Young’s modulus obtained from IET was compared with measurements by the nano-indentation technique. We did not observe any significant difference and the value is in accordance with the one reported in the literature. This work presents a new methodology on the determination of the elasticity constants of thin films using Impulse Excitation Technique. A formulation allowing the determination of the shear modulus of a coating, whatever the thickness, was developed and used to determine the macroscopic elasticity constants of tungsten films. The developed model was validated numerically and experimentally.

Keywords: characterization, coating, dynamical resonant method, Poisson's ratio, PVD, shear modulus, Young's modulus

Procedia PDF Downloads 355

Authors: Irna Malganova

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The paper assessed the effectiveness of the use of urban functional zoning using the method of M.A. Kramer by the example of Kazan city (Republic of Tatarstan, Russian Federation) using geoinformation technologies. On the basis of the data obtained, the calculations were carried out to obtain data on population density, overcoming geographic determinism, as well as the effectiveness of the formation of urban frameworks. The authors proposed recommendations for the effectiveness of municipal frameworks in the period from 2018 to 2021: economic, social, environmental and social. The study of effective territorial planning in a given period allows to display of the dynamics of planning changes, as well as assessment of changes in the formation of urban frameworks. Based on the incoming data obtained from the master plan of the municipal formation of Kazan, in the period from 2018 to 2021, there was an increase in population by 13841 people or 1.1% of the values of 2018. In addition, the area of Kazan increased by 2419.6 hectares. In the structure of the distribution of areas of functional zones, there was an increase in such zones of the municipality as zones of residential and public purpose. Changes in functional zoning, as well as territories requiring reorganization, are presented using geoinformation technologies in open-source software Quantum Geographic Information System (QGIS 3.32). According to the calculations based on the method of functional zoning efficiency by M.A. Kreimer, the territorial-planning structure of Kazan City is quite effective. However, in the development of spatial planning concepts, it is possible to emphasize the weakened interest of the population in the development of territorial planning documents. Thus, the approach to spatial planning of Kazan differs from foreign methods and approaches based on the joint development of planning directions and development of territories of municipalities between the developers of the planning structure, business representatives and the population. The population plays the role of the target audience on which territorial planning is oriented. It follows that there is a need to satisfy the opinions and demands of the population.

Keywords: spatial development, metamorphosis, Kazan city, spatial planning, efficiency, geographic determinism., GIS, QGIS

Procedia PDF Downloads 69

Authors: Qi LI, Xu Lin, Nengming Lin

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Objective: The aim of this study was to investigate the role of desmocollin 2 (DSC2) protein in the proliferation, migration and drug resistance of lung cancer cells. Method: CCK-8 assays and colony formation assays were used to evaluate the effect of dsc2 regulation on cancer cell viability and colony formation. Transwell assays and wound healing assays were also performed. Cell flow double staining was used to detect the apoptosis rate of cells with DSC2, which was added cisplatin. Western blot assay was used to detect cell cycle, PI3k/Akt and apoptosis-related proteins. Results: Our data showed that dsc2 is upregulated in clinical lung cancer tissues compared with pericarcinomatous tissues, and it is differentially expressed in lung cancer cell lines. The down-regulation of dsc2 in A549 and H358 lung cancer cells significantly suppressed the cell proliferation, metastasis, and motility. In contrast, the opposite effects were observed in overexpression of dsc2 both in H23 and PC9 cell lines. In addition to lung adenocarcinoma cell lines, we also examined its expression in lung squamous cell lines, such as H226. Western blotting showed that dsc2 could reduce the level of phosphorylated Akt (Ser 473) and p-mTOR. Thus, it is speculated that dsc2 up-regulation promotes proliferation and invasiveness through activation of the PI3K/AKT pathway. Also, knockdown of dsc2 in A549 and H226 could significantly decreased in the levels of cyclinB and wee1 protein. Additionally, flow cytometry showed that dsc2 knockdown combined with cisplatin could significantly enhance cell apoptosis rate. Conclusion: These data suggest that dsc2 promotes the proliferation and migration of lung cancer cells in vitro. Also, the results suggested that dsc2 could affect the cell cycle and apoptosis of lung cells. Furthermore, knockdown of dsc2 could sensitize cisplatin in both lung adenocarcinoma and lung squamous cell lines. Thus we suggested that dsc2 can be used as a therapeutic target for lung cancer.

Keywords: desmocollin 2, cisplatin, lung cancer, PI3K/AKT, lung squamous cell

Procedia PDF Downloads 67

Authors: Anil Koklu, Amin Mansoorifar, Ali Beskok

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Dielectric spectroscopy (DS) is a noninvasive, label free technique for a long term real-time measurements of the impedance spectra of biological cells. DS enables characterization of cellular dielectric properties such as membrane capacitance and cytoplasmic conductivity. We have developed a lab-on-a-chip device that uses an electro-activated microwells array for loading, DS measurements, and unloading of biological cells. We utilized from dielectrophoresis (DEP) to capture target cells inside the wells and release them after DS measurement. DEP is a label-free technique that exploits differences among dielectric properties of the particles. In detail, DEP is the motion of polarizable particles suspended in an ionic solution and subjected to a spatially non-uniform external electric field. To the best of our knowledge, this is the first microfluidic chip that combines DEP and DS to analyze biological cells using electro-activated wells. Device performance is tested using two different cell lines of prostate cancer cells (RV122, PC-3). Impedance measurements were conducted at 0.2 V in the 10 kHz to 40 MHz range with 6 s time resolution. An equivalent circuit model was developed to extract the cell membrane capacitance and cell cytoplasmic conductivity from the impedance spectra. We report the time course of the variations in dielectric properties of PC-3 and RV122 cells suspended in low conductivity medium (LCB), which enhances dielectrophoretic and impedance responses, and their response to sudden pH change from a pH of 7.3 to a pH of 5.8. It is shown that microfluidic chip allowed online measurements of dielectric properties of prostate cancer cells and the assessment of the cellular level variations under external stimuli such as different buffer conductivity and pH. Based on these data, we intend to deploy the current device for single cell measurements by fabricating separately addressable N × N electrode platforms. Such a device will allow time-dependent dielectric response measurements for individual cells with the ability of selectively releasing them using negative-DEP and pressure driven flow.

Keywords: microfluidic, microfabrication, lab on a chip, AC electrokinetics, dielectric spectroscopy

Procedia PDF Downloads 139

Authors: Nishanthi N. S., Srikanth Vedantam

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Intracellular delivery of materials has always proved to be a challenge in research and therapeutic applications. Usually, vector-based methods, such as liposomes and polymeric materials, and physical methods, such as electroporation and sonoporation have been used for introducing nucleic acids or proteins. Reliance on exogenous materials, toxicity, off-target effects was the short-comings of these methods. Microinjection was an alternative process which addressed the above drawbacks. However, its low throughput had hindered its adoption widely. Mechanical deformation of cells by squeezing them through constriction channel can cause the temporary development of pores that would facilitate non-targeted diffusion of materials. Advantages of this method include high efficiency in intracellular delivery, a wide choice of materials, improved viability and high throughput. This cell squeezing process can be studied deeper by employing simple models and efficient computational procedures. In our current work, we present a finite sized dissipative particle dynamics (FDPD) model to simulate the dynamics of the cell flowing through a constricted channel. The cell is modeled as a capsule with FDPD particles connected through a spring network to represent the membrane. The total energy of the capsule is associated with linear and radial springs in addition to constraint of the fixed area. By performing detailed simulations, we studied the strain on the membrane of the capsule for channels with varying constriction heights. The strain on the capsule membrane was found to be similar though the constriction heights vary. When strain on the membrane was correlated to the development of pores, we found higher porosity in capsule flowing in wider channel. This is due to localization of strain to a smaller region in the narrow constriction channel. But the residence time of the capsule increased as the channel constriction narrowed indicating that strain for an increased time will cause less cell viability.

Keywords: capsule, cell squeezing, dissipative particle dynamics, intracellular delivery, microfluidics, numerical simulations

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

Procedia PDF Downloads 307