Search results for: geometrically nonlinear analysis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 27605

Search results for: geometrically nonlinear analysis

5465 Development of a Regression Based Model to Predict Subjective Perception of Squeak and Rattle Noise

Authors: Ramkumar R., Gaurav Shinde, Pratik Shroff, Sachin Kumar Jain, Nagesh Walke

Abstract:

Advancements in electric vehicles have significantly reduced the powertrain noise and moving components of vehicles. As a result, in-cab noises have become more noticeable to passengers inside the car. To ensure a comfortable ride for drivers and other passengers, it has become crucial to eliminate undesirable component noises during the development phase. Standard practices are followed to identify the severity of noises based on subjective ratings, but it can be a tedious process to identify the severity of each development sample and make changes to reduce it. Additionally, the severity rating can vary from jury to jury, making it challenging to arrive at a definitive conclusion. To address this, an automotive component was identified to evaluate squeak and rattle noise issue. Physical tests were carried out for random and sine excitation profiles. Aim was to subjectively assess the noise using jury rating method and objectively evaluate the same by measuring the noise. Suitable jury evaluation method was selected for the said activity, and recorded sounds were replayed for jury rating. Objective data sound quality metrics viz., loudness, sharpness, roughness, fluctuation strength and overall Sound Pressure Level (SPL) were measured. Based on this, correlation co-efficients was established to identify the most relevant sound quality metrics that are contributing to particular identified noise issue. Regression analysis was then performed to establish the correlation between subjective and objective data. Mathematical model was prepared using artificial intelligence and machine learning algorithm. The developed model was able to predict the subjective rating with good accuracy.

Keywords: BSR, noise, correlation, regression

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5464 Mouse Knockouts for Elucidating the Role of Cysteine-Rich Angiogenic Inducer 61 in Tendon Development and Maintenance

Authors: Josephine Hai, Jie Jiang, Karen M. Lyons

Abstract:

Of the musculoskeletal tissues, tendon is least understood in terms of biological development. The current study examines Cysteine-rich angiogenic inducer 61, or CCN1, a member of the CCN family of secreted matricellular proteins that regulate cell behavior via intercellular signaling. Though CCN1 is notable in limiting fibrosis by inducing senescence in fibroblasts, little is known about its role in normal fibrous tissue, where it may be essential to the development of ECM-rich structures like tendons. We found that CCN1 knockout mice (using limb-specific Prx1-Cre) exhibited clubfoot and waddling gaits, a unique phenotype not described in any other mutant to date. Histological analysis showed that the Achilles and patellar tendons, where we previously found high CCN1 expression in adult reporter mice, were thicker and denser in the Prx1-Cre knockouts than in their wildtype littermates. We then hypothesized that CCN1 is required directly in tendon progenitor cells for normal tendon development and generated tendon-specific CCN1 knockout mice using Scx-Cre. We observed similar Achilles/patellar tendon morphology among the Scx-Cre and Prx1-Cre mutants, indicating that the phenotype is a direct result of CCN1’s loss in tendon. To further study phenotype onset and progression, we will histologically characterize these tendons across different developmental time-points. We will also perform RNA-seq and qPCR to analyze tenocyte gene expression and expect fibrotic marker upregulation in the Scx-Cre mutants if CCN1 is required to maintain a normal tendon phenotype. Thus, our study aims to elucidate the molecular mechanisms underlying tendon formation and maintenance. Understanding tendons at the most basic level invites novel approaches to tendon repair.

Keywords: development, matricellular, musculoskeletal, tendon

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5463 The American Theater: Latinos Performing as American Citizens by Supporting Trump's Ideals

Authors: Mariana Anaya Villafana

Abstract:

The sudden change of a significant percentage of the Latino community in the United States elections towards a Republican political orientation was reflected during the 2016 presidential election. This moment represented a radical change that is happening inside the Latino community in the United States, the support they have given to Trump's campaign only demonstrates their support for new anti-immigration regulations and conservative values, which are causing a division of ideologies inside the Latino community. One of the main goals of the following research is to understand the whole phenomenon 'Why would people join their own oppressor?' Align themselves with the politics that prevent many of their relatives to come to the United States and made the assimilation process difficult for their parents. It is important to prove that a change in the identity has happened, through the use of power relations and the attachment to the desired object. A group of Hispanics/Latinos have decided to vote for Trump in order to belong to a society that hasn’t been able to fully include them within it, an action that can result on the non-intentional harm of the values and aims of the rest of the Latino/Hispanic community. In order to understand their new political beliefs, it is necessary to use the method of discourse analysis to comprehend those comments and interviews that are published on web sites such as: 'Latinos for Trump' and 'GOP Hispanic Division'. Among the results that the research has shown, the notion of the 'American Dream' can be considered as a determinant object for the construction of a new identity that is rooted in hard work and legality. One that is proud of the Latino heritage but still wants to maintain the boundaries between legality and illegality in relation to the immigrants. This discourse results on a contradiction to most of the cases because they mention that their families came to the U.S. as immigrants; the only difference is that they work hard to obtain legal citizenship.

Keywords: populism, identity, Latino Community, migration

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5462 Experimental Analysis for the Inlet of the Brazilian Aerospace Vehicle 14-X B

Authors: João F. A. Martos, Felipe J. Costa, Sergio N. P. Laiton, Bruno C. Lima, Israel S. Rêgo, Paulo P. G. Toro

Abstract:

Nowadays, the scramjet is a topic that has attracted the attention of several scientific communities (USA, Australia, Germany, France, Japan, India, China, Russia), that are investing in this in this type of propulsion system due its interest to facilitate access to space and reach hypersonic speed, who have invested in this type of propulsion due to the interest in facilitating access to space. The Brazilian hypersonic scramjet aerospace vehicle 14-X B is a technological demonstrator of a hypersonic airbreathing propulsion system based on the supersonic combustion (scramjet) intended to be tested in flight into the Earth's atmosphere at 30 km altitude and Mach number 7. The 14-X B has been designed at the Prof. Henry T. Nagamatsu Laboratory of Aerothermodynamics and Hypersonics of the Institute for Advanced Studies (IEAv) in Brazil. The IEAv Hypersonic Shock Tunnel, named T3, is a ground-test facility able to reproduce the flight conditions as the Mach number as well as pressure and temperature in the test section close to those encountered during the test flight of the vehicle 14-X B into design conditions. A 1-m long stainless steel 14-X B model was experimentally investigated at T3 Hypersonic Shock Tunnel, for freestream Mach number 7. Static pressure measurements along the lower surface of the 14-X B model, along with high-speed schlieren photographs taken from the 5.5° leading edge and the 14.5° deflection compression ramp, provided experimental data that were compared to the analytical-theoretical solutions and the computational fluid dynamics (CFD) simulations. The results show a good qualitative agreement, and in consequence demonstrating the importance of these methods in the project of the 14-X B hypersonic aerospace vehicle.

Keywords: 14-X, CFD, hypersonic, hypersonic shock tunnel, scramjet

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5461 Category-Base Theory of the Optimum Signal Approximation Clarifying the Importance of Parallel Worlds in the Recognition of Human and Application to Secure Signal Communication with Feedback

Authors: Takuro Kida, Yuichi Kida

Abstract:

We show a base of the new trend of algorithm mathematically that treats a historical reason of continuous discrimination in the world as well as its solution by introducing new concepts of parallel world that includes an invisible set of errors as its companion. With respect to a matrix operator-filter bank that the matrix operator-analysis-filter bank H and the matrix operator-sampling-filter bank S are given, firstly, we introduce the detailed algorithm to derive the optimum matrix operator-synthesis-filter bank Z that minimizes all the worst-case measures of the matrix operator-error-signals E(ω) = F(ω) − Y(ω) between the matrix operator-input-signals F(ω) and the matrix operator-output signals Y(ω) of the matrix operator-filter bank at the same time. Further, feedback is introduced to the above approximation theory and it is indicated that introducing conversations with feedback does not superior automatically to the accumulation of existing knowledge of signal prediction. Secondly, the concept of category in the field of mathematics is applied to the above optimum signal approximation and is indicated that the category-based approximation theory is applied to the set-theoretic consideration of the recognition of humans. Based on this discussion, it is shown naturally why the narrow perception that tends to create isolation shows an apparent advantage in the short term and, often, why such narrow thinking becomes intimate with discriminatory action in a human group. Throughout these considerations, it is presented that, in order to abolish easy and intimate discriminatory behavior, it is important to create a parallel world of conception where we share the set of invisible error signals, including the words and the consciousness of both worlds.

Keywords: signal prediction, pseudo inverse matrix, artificial intelligence, conditional optimization

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5460 Clinical Parameters Response to Low Level Laser Versus Monochromatic Near Infrared Photo Energy in Diabetic Patient with Peripheral Neuropathy

Authors: Abeer Ahmed Abdehameed

Abstract:

Background: Diabetic sensorimotor polyneuropathy (DSP) is one of the most common micro vascular complications of type 2 diabetes. Loss of sensation is thought to contribute to lake of static and dynamic stability and increased risk of falling. Purpose: The purpose of this study was to compare the effects of low level laser (LLL) and monochromatic near infrared photo energy (MIRE) on pain , cutaneous sensation, static stability and index of lower limb blood flow in diabetic with peripheral neuropathy. Methods: Forty subjects with diabetic peripheral neuropathy were recruited for study. They were divided into two groups: The ( MIRE) group that included (20) patients and (LLL) group included (20) patients. All patients in the study had been subjected to various physical assessment procedures including pain, cutaneous sensation, Doppler flow meter and static stability assessments. The baseline measurements were followed by treatment sessions that conducted twice a week for 6 successive weeks. Results: The statistical analysis of the data had revealed significant improvement of the pain in both groups, with significant improvement in cutaneous sensation and static balance in (MIRE) group compared to (LLL) group; on the other hand results showed no significant differences on lower limb blood flow in both groups. Conclusion: Low level laser and monochromatic near infrared therapy can improve painful symptoms in patients with diabetic neuropathy. On the other hand (MIRE) is useful in improving cutaneous sensation and static stability in patients with diabetic neuropathy.

Keywords: diabetic neuropathy, doppler flow meter, low level laser, monochromatic near infrared photo energy

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5459 Digital Self-Identity and the Role of Interactivity in Psychiatric Assessment and Treatment

Authors: Kevin William Taylor

Abstract:

This work draws upon research in the fields of games development and mental health treatments to assess the influence that interactive entertainment has on the populous, and the potential of technology to affect areas of psychiatric assessment and treatment. It will use studies to establish the evolving direction of interactive media in the development of ‘digital self-identity,’ and how this can be incorporated into treatment to the benefit of psychiatry. It will determine that this approach will require collaborative production between developers and psychiatrists in order to ensure precise goals are met, improving the success of serious gaming for psychiatric assessment and treatment. Analysis documents the reach of video games across a growing global community of gamers, highlighting cases of the positives and negatives of video game usage. The games industry is largely oblivious to the psychological negatives, with psychiatrists encountering new conditions such as gaming addiction, which is now recognized by the World Health Organization. With an increasing amount of gamers worldwide, and an additional time per day invested in online gaming and character development, the concept of virtual identity as a means of expressing the id needs further study to ensure successful treatment. In conclusion, the assessment and treatment of game-related conditions are currently reactionary, and while some mental health professionals have begun utilizing interactive technologies to assist with the assessment and treatment of conditions, this study will determine how the success of these products can be enhanced. This will include collaboration between software developers and psychiatrists, allowing new avenues of skill-sharing in interactive design and development. Outlining how to innovate approaches to engagement will reap greater rewards in future interactive products developed for psychiatric assessment and treatment.

Keywords: virtual reality, virtual identity, interactivity, psychiatry

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5458 Experiences of Homophobia, Machismo and Misogyny in Tourist Destinations: A Netnography in a Facebook Community of LGBT Backpackers

Authors: Renan De Caldas Honorato, Ana Augusta Ferreira De Freitas

Abstract:

Homosexuality is still criminalized in a large number of countries. In some of them, being gay or lesbian can even be punished by death. Added to this context, the experiences of social discrimination faced by the LGBT population, including homophobia, machismo and misogyny, cause numerous restrictions throughout their lives. The possibility of confronting these challenges in moments that should be pleasant, such as on a trip or on vacation, is unpleasant, to say the least. In the current scenario of intensifying the use of Social network sites (SNSs) to search for information, including in the tourist area, this work aims to analyze the sharing of tourist experiences with situations of confrontation and perceptions of homophobia, machismo and misogyny, and restrictions suffered in tourist destinations. The fieldwork is a community of LGBT backpackers based on Facebook. Netnography was the core method adopted. A qualitative approach was conducted and 463 publications posted from January to December 2020 were assessed through the computer-mediated discourse analysis (CMDA). The results suggest that these publications exist to identify the potential exposure to these offensive behaviors while traveling. Individuals affirm that the laws, positive or not, in relation to the LGBT public are not the only factors for a place to be defined as safe or not for gay travelers. The social situation of a country and its laws are quite different and this is the main target of these publications. The perception of others about the chosen destination is more important than knowing your rights and the legal status of each country and it also lessens uncertainty, even when they are never totally confident when choosing a travel destination. In certain circumstances, sexual orientation also needs to be protected from the judgment of hosts and residents. The systemic treatment of homophobic behavior and the construction of a more inclusive society are urgent.

Keywords: homophobia, hospitality, machismo, misogyny

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5457 The New Insight about Interspecies Transmission of Iranian H9N2 Influenza Viruses from Avian to Human

Authors: Masoud Soltanialvar, Ali Bagherpour

Abstract:

Documented cases of human infection with H9N2 avian influenza viruses, first detected in 1999 in Hong Kong and China, indicate that these viruses can be directly transmitted from birds to humans. In this study, we characterized the mutation in the Hemagglutinin (HA) genes and proteins that correlates with a shift in affinity of the Hemagglutinin (HA) protein from the “avian” type sialic receptors to the “human” type in 10 Iranian isolates. We delineated the genomes and receptor binding profile of HA gene of some field isolates and established their phylogenetic relationship to the other Asian H9N2 sub lineages. A total of 1200 tissue samples collected from 40 farms located in various states of Iran during 2008 – 2010 as part of a program to monitor Avian Influenza Viruses (AIV) infection. To determine the genetic relationship of Iranian viruses, the Hemagglutinin (HA) genes from ten isolates were amplified and sequenced (by RT-PCR method). Nucleotide sequences (orf) of the (HA) genes were used for phylogenetic tree construction. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) in all ten Iranian isolates which indicates a preference to binding of α (2–6) sialic acid receptors, so these Iranian H9N2 viruses have the potential to infect human beings. These isolates showed high degree of homology with 2 human H9N2 isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of showed that all the HA genes of the Iranian H9N2 viruses fall into a single group within a G1-like sublineage which had contributed as donor of six internal genes to H5N1 highly pathogenic avian influenza. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.

Keywords: influenza virus, hemagglutinin, neuraminidase, Iran

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5456 Features of the Functional and Spatial Organization of Railway Hubs as a Part of the Urban Nodal Area

Authors: Khayrullina Yulia Sergeevna, Tokareva Goulsine Shavkatovna

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The article analyzes the modern major railway hubs as a main part of the Urban Nodal Area (UNA). The term was introduced into the theory of urban planning at the end of the XX century. Tokareva G.S. jointly with Gutnov A.E. investigated the structure-forming elements of the city. UNA is the basic unit, the "cell" of the city structure. Specialization is depending on the position in the frame or the fabric of the city. This is related to feature of its organization. Spatial and functional features of UNA proposed to investigate in this paper. The base object for researching are railway hubs as connective nodes of inner and extern-city communications. Research used a stratified sampling type with the selection of typical objects. Research is being conducted on the 14 railway hubs of the native and foreign experience of the largest cities with a population over 1 million people located in one and close to the Russian climate zones. Features of the organization identified in the complex research of functional and spatial characteristics based on the hypothesis of the existence of dual characteristics of the organization of urban nodes. According to the analysis, there is using the approximation method that enable general conclusions of a representative selection of the entire population of railway hubs and it development’s area. Results of the research show specific ratio of functional and spatial organization of UNA based on railway hubs. Based on it there proposed typology of spaces and urban nodal areas. Identification of spatial diversity and functional organization’s features of the greatest railway hubs and it development’s area gives an indication of the different evolutionary stages of formation approaches. It help to identify new patterns for the complex and effective design as a prediction of the native hub’s development direction.

Keywords: urban nodal area, railway hubs, features of structural, functional organization

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5455 Relationship between Employee Welfare Practices and Performance of Non-Governmental Organizations in Kenya

Authors: Protus A. Lumiti, Susan O. Wekesa, Mary Omondi

Abstract:

Performance is a key pillar to the accomplishment of the goals of all organizations, whether private, public or non- profit. Employees are the intellectual assets of the organization and they are an avenue to the achievement of competitive advantage. An employee welfare service in an organization is vital in fostering employee motivation and improving their productivity. In view of this, the main goal of this research was to determine the relationship between employee welfare practices and the performance of non-governmental organizations in Kenya. The study was guided by four objectives, namely: to establish, determine, evaluate and assess the relationship between employee welfare practices and the performance of non-governmental organizations in Kenya. The study utilized a survey design using both qualitative and quantitative approaches. In this study, a purposive, stratified and simple random sampling technique was used to arrive at a sample of 355 respondents who comprised senior managers, middle level managers and operational employees out of the targeted population of 14,283 employees of non-governmental organizations working in Nairobi County. The primary data collection tools were questionnaires supplemented by an interview schedule, while secondary data was obtained from reviewed journals, published books and articles. Data analysis was done using Statistical Packages for Social Sciences Software version 23. The study utilized multiple linear regression and a structural equation model. The findings of the study were that: employee welfare practices had a positive and significant relationship with the performance of Non-governmental organizations in Kenya. In addition, there was also a linear relationship between the independent variables and the dependent variable and the study concluded that there was a relationship between the predictor variable and the dependent variable of the study. The study recommended that management of No-governmental organization boards in Kenya should come up with a comprehensive policy document on employee welfare practices in order to enhance the performance of non-governmental organizations in Kenya.

Keywords: employee, economic, performance, welfare

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5454 Exploring the Effect of Using Lesh Model in Enhancing Prospective Mathematics Teachers’ Number Sense

Authors: Areej Isam Barham

Abstract:

Developing students’ number sense is an essential element in the learning of mathematics. Number sense is one of the foundational ideas in mathematics where students need to understand numbers, representing them in different ways, and realize the relationships among numbers. Number sense also reflects students’ understanding of the meaning of operations, how they related to one another, how to compute fluently and make reasonable estimates. Developing students’ number sense in the mathematics classroom requires good preparation for mathematics teachers, those who will direct their students towards the real understanding of numbers and its implementation in the learning of mathematics. This study describes the development of elementary prospective mathematics teachers’ number sense through a mathematics teaching methods course at Qatar University. The study examined the effect of using the Lesh model in enhancing mathematics prospective teachers’ number sense. Thirty-nine elementary prospective mathematics teachers involved in the current study. The study followed an experimental research approach, and quantitative research methods were used to answer the research questions. Pre-post number sense test was constructed and implemented before and after teaching by using the Lesh model. Data were analyzed using Statistical Packages for Social Sciences (SPSS). Descriptive data analysis and t-test were used to examine the impact of using the Lesh model in enhancing prospective teachers’ number sense. Finding of the study indicated poor number sense and limited numeracy skills before implementing the use of the Lesh model, which highly demonstrate the importance of the study. The results of the study also revealed a positive impact on the use of the Lesh model in enhancing prospective teachers’ number sense with statistically significant differences. The discussion of the study addresses different features and issues related to the participants’ number sense. In light of the study, the research presents recommendations and suggestions for the future development of mathematics prospective teachers’ number sense.

Keywords: number sense, Lesh model, prospective mathematics teachers, development of number sense

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5453 Queerness and Gender Representation Through the Lens of Five Ghanaian Artists

Authors: Sela Adjei

Abstract:

This research delves into the nuanced representations of queerness in Ghana, presented through photographs, illustrations, film and music videos on social media and streaming platforms. The study focuses on the works of five Ghanaian artists (Va-Bene Elikem Fiatsi, Angel Maxine, Josephine Kuuire, Bright Ackwerh and Philip Nee Whang) within the context of Ghana's evolving media landscape. Of primary concern is a need to uncover the various aspects of queerness captured within the distinct artistic expressions of these five creatives. This study adopts a qualitative approach by analyzing artistic expressions of queerness in Ghana’s digital media spaces. Content analysis and visual semiotics served as the guiding tools to discuss and decipher the nuanced messages embedded in their works, considering both the visual and narrative aspects. This dual approach takes into account both the visual aesthetics and narrative elements, enhancing our understanding of the complex interplay between queerness and gender representation in the media. This study's contribution is twofold. First, it enriches the discourse surrounding queerness as portrayed by artists within Ghana's vibrant media landscape and situates their works within the broader discourse of global gender identities. Secondly, analyzing the creative output of these five Ghanaian artists broadens our understanding of gender minorities and the various challenges they face in Ghana (currently debating in parliament to pass an anti-LGBTQ+ bill that criminalizes activities related to gender minority groups). While focusing on the intersection of queerness, art, and gender identities, the reflections in this study challenge existing narratives and offer fresh insights into how these artists navigate and challenge societal norms through their creative expressions.

Keywords: queer, film, representation, streaming, media, gender

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5452 Surface Thermodynamics Approach to Mycobacterium tuberculosis (M-TB) – Human Sputum Interactions

Authors: J. L. Chukwuneke, C. H. Achebe, S. N. Omenyi

Abstract:

This research work presents the surface thermodynamics approach to M-TB/HIV-Human sputum interactions. This involved the use of the Hamaker coefficient concept as a surface energetics tool in determining the interaction processes, with the surface interfacial energies explained using van der Waals concept of particle interactions. The Lifshitz derivation for van der Waals forces was applied as an alternative to the contact angle approach which has been widely used in other biological systems. The methodology involved taking sputum samples from twenty infected persons and from twenty uninfected persons for absorbance measurement using a digital Ultraviolet visible Spectrophotometer. The variables required for the computations with the Lifshitz formula were derived from the absorbance data. The Matlab software tools were used in the mathematical analysis of the data produced from the experiments (absorbance values). The Hamaker constants and the combined Hamaker coefficients were obtained using the values of the dielectric constant together with the Lifshitz equation. The absolute combined Hamaker coefficients A132abs and A131abs on both infected and uninfected sputum samples gave the values of A132abs = 0.21631x10-21Joule for M-TB infected sputum and Ã132abs = 0.18825x10-21Joule for M-TB/HIV infected sputum. The significance of this result is the positive value of the absolute combined Hamaker coefficient which suggests the existence of net positive van der waals forces demonstrating an attraction between the bacteria and the macrophage. This however, implies that infection can occur. It was also shown that in the presence of HIV, the interaction energy is reduced by 13% conforming adverse effects observed in HIV patients suffering from tuberculosis.

Keywords: absorbance, dielectric constant, hamaker coefficient, lifshitz formula, macrophage, mycobacterium tuberculosis, van der waals forces

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5451 Extraskeletal Ewing Sarcoma- Experience in a Tertiary Cancer Care Centre of India

Authors: Himanshu Rohela

Abstract:

BACKGROUND: Ewing sarcoma can arise in either bone or soft tissue. Extraskeletal Ewing sarcoma (EES) is an uncommon primary tumor of the soft tissues, accounting for 20 30% of all reported cases of ES. AIM: Was to investigate demographic distribution, survival analysis and factors affecting the survival and recurrence in patients of EES. METHODS: Retrospective study of 19 biopsy-proven EES was performed. Overall survival (OS) using log-rank test and factors affecting OS and local recurrence (LR) were evaluated for the entire cohort. RESULTS: Patients with EES had a mean age of 19.5 and it was more commonly seen in males (63%). Axial location (58%) and solitary presentation (84%) were more common. The average size was 11 cm, 3 of 19 were metastatic at presentation, with the lung beings the most common site for metastasis. 17 received NACT, 16 with VAC-IE regimen and 1 underwent a second line with GEM/DOCE regimen. Unplanned surgery was done in 2 of 19. 3 patients received definitive RT and 13 underwent surgical-wide local excision. 2 of 13 showed good response to NACT. 10 patients required readmission out of which 6 patients had chemotherapy-related complications, 2 had surgical site complications and one patient developed secondary AML post-completion of treatment. A total of 4 patients had a recurrence. One had local recurrence alone, one had distant recurrence alone and 2 patients had a distant and local recurrence both. Tumor size >10 cm, axial location, and previous unplanned surgery was associated with higher LR rate. The mean overall survival was 32 months (2.66 years), with higher rates seen in non-metastatic and non-recurrent settings. CONCLUSIONS: Early and accurate diagnosis is the key to the management of EES, with promising results seen via NACT and RO resection regimens. But further studies with larger study groups are needed to standardize the treatment protocol and evaluate its efficacy.

Keywords: Ewings, sarcoma, extraskeletal, chemotherapy

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5450 Design & Development of a Static-Thrust Test-Bench for Aviation/UAV Based Piston Engines

Authors: Syed Muhammad Basit Ali, Usama Saleem, Irtiza Ali

Abstract:

Internal combustion engines have been pioneers in the aviation industry, use of piston engines for aircraft propulsion, from propeller-driven bi-planes to turbo-prop, commercial, and cargo airliners. To provide an adequate amount of thrust piston engine rotates the propeller at a specific rpm, allowing enough mass airflow. Thrust is the only forward-acting force of an aircraft that helps heavier than air bodies to fly, depending on the mathematical model and variables included in that with the correct measurement. Test-benches have been a bench-mark in the aerospace industry to analyse the results before a flight, having paramount significance in reliability and safety engineering, depending on the mathematical model and variables included in that with the correct measurement. Calculation of thrust from a piston engine also depends on environmental changes, the diameter of the propeller, and the density of air. The project would be centered on piston engines used in the aviation industry for light aircraft and UAVs. A static thrust test bench involves various units, each performing a designed purpose to monitor and display. Static thrust tests are performed on the ground, and safety concerns hold paramount importance. The execution of this study involves research, design, manufacturing, and results based on reverse engineering initiating from virtual design, analytical analysis, and simulations. The final evaluation of results gathered from various methods such as co-relation between conventional mass-spring and digital loadcell. On average, we received 17.5kg of thrust (25+ engine run-ups – around 40 hours of engine run), only 10% deviation from analytically calculated thrust –providing 90% accuracy.

Keywords: aviation, aeronautics, static thrust, test bench, aircraft maintenance

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5449 Exploring Simple Sequence Repeats within Conserved microRNA Precursors Identified from Tea Expressed Sequence Tag (EST) Database

Authors: Anjan Hazra, Nirjhar Dasgupta, Chandan Sengupta, Sauren Das

Abstract:

Tea (Camellia sinensis) has received substantial attention from the scientific world time to time, not only for its commercial importance, but also for its demand to the health-conscious people across the world for its extensive use as potential sources of antioxidant supplement. These health-benefit traits primarily rely on some regulatory networks of different metabolic pathways. Development of microsatellite markers from the conserved genomic regions is being worthwhile for studying the genetic diversity of closely related species or self-pollinated species. Although several SSR markers have been reported, in tea the trait-specific Simple Sequence Repeats (SSRs) are yet to be identified, which can be used for marker assisted breeding technique. MicroRNAs are endogenous, noncoding, short RNAs directly involved in regulating gene expressions at the post-transcriptional level. It has been found that diversity in miRNA gene interferes the formation of its characteristic hair pin structure and the subsequent function. In the present study, the precursors of small regulatory RNAs (microRNAs) has been fished out from tea Expressed Sequence Tag (EST) database. Furthermore, the simple sequence repeat motifs within the putative miRNA precursor genes are also identified in order to experimentally validate their existence and function. It is already known that genic-SSR markers are very adept and breeder-friendly source for genetic diversity analysis. So, the potential outcome of this in-silico study would provide some novel clues in understanding the miRNA-triggered polymorphic genic expression controlling specific metabolic pathways, accountable for tea quality.

Keywords: micro RNA, simple sequence repeats, tea quality, trait specific marker

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5448 A Review of Current Practices in Tattooing of Colonic Lesion at Endoscopy

Authors: Dhanashree Moghe, Roberta Bullingham, Rizwan Ahmed, Tarun Singhal

Abstract:

Aim: The NHS Bowel Screening Programme recommends the use of endoscopic tattooing for suspected malignant lesions that later require surgical or endoscopic localisation, using local protocols as guidance. This is in accordance with guidance from the BSG (The British Society of Gastroenterologists). We used a well-recognised local protocol as a standard to audit current tattooing practice in a large district general hospital with no current local guidelines. Method: A retrospective quantitative analysis of 50 patients who underwent segmental colonic resection for cancer over a 6-month period in 2021. We reviewed historic electronic endoscopy reports recording relevant data on tattoo indication and placement. Secondly, we carried out an anonymous survey of 16 independent lower GI endoscopists on self-reported details of their practice. Results: In our study, 28 patients (56%) had a tattoo placed at the time of their colonoscopy. Of these, only 53% (n=15) had the tattoo distal to the lesion, with the measured distance of the tattoo from the lesion only being documented in 8 reports. Only seven patients (25%) had a circumferential (4 quadrant) placement of the tattoo. 13 patients had lesions either in the caecum or rectum, locations deemed unnecessary as per BSG guidelines. Of the survey responses collected, there were four different protocols being used to guide practice. Only 50% of respondents placed tattoos at the correct distance from the lesion, and 83% placed the correct number of tattoos. Conclusion: There is a lack of standardisation of practices in colonic tattooing demonstrated in our study with incomplete compliance to our standard. The inadequate documentation of tattoo location can contribute to confusion and inaccuracy in the intraoperative localisation of lesions. This has the potential to increase operation length and morbidity. There is a need to standardise both technique and documentation in colonoscopic tattooing practice.

Keywords: colorectal cancer, endoscopic tattooing, colonoscopy, NHS BSCP

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5447 Conserving History: Evaluating and Selecting Effective Restoration Methods for a Fragment Mural Painting from Amarna

Authors: Kholod Khairy Salama, Shabban Hassan Thabet

Abstract:

In the present study, a comprehensive investigation has been undertaken into an Egyptian mural painting with feet wear slippers approach to choose the most successful restoration methods. The mural painting under examination dates back to the Amarna period; it was detached from a wall of an unknown tomb in Egypt, and currently, it is initially displayed in a showcase at the Egyptian Museum – Tahrir Square – Cairo, Egypt. The main objectives of this research were to (a) reveal the pigment used in the mural painting, (b) reveal the medium used with colours, (c) determine the technique of manufacturing, (e) determine the ground support, and (f) reveal the main deterioration aspects. The analytical techniques used for investigation were Optical Microscopy, Raman, X-ray Florescence, X-ray diffraction, and Fourier transform infrared coupled with attenuated total reflectance “FTIR-ATR”. The investigation revealed that the vital deterioration factors affecting the object. This research aims to examine and analyze the mural painting to choose the suitable method for the restoration process (a) define the colours through comparative analysis to choose the suitable material for cleaning, (b) define the natural structure of the ground support layer, which appeared as mud layer (c) determine the medium used with colours (d) diagnosis the presence of the white wash layer, and (e) choose the suitable restoration methods according to the results. Conclusion: This study focused mainly on the physical and chemical properties of the mural painting compound and the main changes that happened to the mural painting material, which caused deterioration and fall down of the painting parts, so we can find the best and optimum restoration ways for this object.

Keywords: mural paintings, Tal Al-Amarna, digital microscope, Raman, XRF, XRD, FTIR

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5446 High Expression Levels and Amplification of rRNA Genes in a Mentally Retarded Child with 13p+: A Familial Case Study

Authors: Irina S. Kolesnikova, Alexander A. Dolskiy, Natalya A. Lemskaya, Yulia V. Maksimova, Asia R. Shorina, Alena S. Telepova, Alexander S. Graphodatsky, Dmitry V. Yudkin

Abstract:

A cytogenetic and molecular genetic study of the family with a male child who had mental retardation and autistic features revealed an abnormal chromosome 13 bearing an enlarged p-arm with amplified ribosomal DNA (rDNA) in a boy and his father. Cytogenetic analysis using standard G-banding and FISH with labeled rDNA probes revealed an abnormal chromosome 13 with an enlarged p-arms due to rDNA amplification in a male child, who had clinically confirmed mental retardation and an autistic behavior. This chromosome is evidently inherited from the father, who has morphologically the same chromosome, but is healthy. The karyotype of the mother was normal. Ag-NOR staining showed brightly stained large whole-p-arm nucleolus organizer regions (NORs) in a child and normal-sized NORs in his father with 13p+-NOR-amount mosaicism. qRT-PCR with specific primers showed highly increased levels of 18S, 28S and 5,8 S ribosomal RNA (rRNA) in the patient’s blood samples compared to a normal healthy control donor. Both patient’s father and mother had no elevated levels of rRNAs expression. Thus, in this case, rRNA level seems to correlate with mental retardation in familial individuals with 13p+. Our findings of rRNA overexpression in a patient with mental retardation and his parents may show a possible link between the karyotype (p-arm enlargement due to rDNA amplification), rDNA functionality (rRNA overexpression), functional changes in the brain and mental retardation. The study is supported by Russian Science Foundation Grant 15-15-10001.

Keywords: mental retardation, ribosomal DNA–rDNA, ribosomal RNA–rRNA, nucleolus organizer region–NOR, chromosome 13

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5445 Elaboration and Validation of a Survey about Research on the Characteristics of Mentoring of University Professors’ Lifelong Learning

Authors: Nagore Guerra Bilbao, Clemente Lobato Fraile

Abstract:

This paper outlines the design and development of the MENDEPRO questionnaire, designed to analyze mentoring performance within a professional development process carried out with professors at the University of the Basque Country, Spain. The study took into account the international research carried out over the past two decades into teachers' professional development, and was also based on a thorough review of the most common instruments used to identify and analyze mentoring styles, many of which fail to provide sufficient psychometric guarantees. The present study aimed to gather empirical data in order to verify the metric quality of the questionnaire developed. To this end, the process followed to validate the theoretical construct was as follows: The formulation of the items and indicators in accordance with the study variables; the analysis of the validity and reliability of the initial questionnaire; the review of the second version of the questionnaire and the definitive measurement instrument. Content was validated through the formal agreement and consensus of 12 university professor training experts. A reduced sample of professors who had participated in a lifelong learning program was then selected for a trial evaluation of the instrument developed. After the trial, 18 items were removed from the initial questionnaire. The final version of the instrument, comprising 33 items, was then administered to a sample group of 99 participants. The results revealed a five-dimensional structure matching theoretical expectations. Also, the reliability data for both the instrument as a whole (.98) and its various dimensions (between .91 and .97) were very high. The questionnaire was thus found to have satisfactory psychometric properties and can therefore be considered apt for studying the performance of mentoring in both induction programs for young professors and lifelong learning programs for senior faculty members.

Keywords: higher education, mentoring, professional development, university teaching

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5444 Isolation and Characterisation of Novel Environmental Bacteriophages Which Target the Escherichia coli Lamb Outer Membrane Protein

Authors: Ziyue Zeng

Abstract:

Bacteriophages are viruses which infect bacteria specifically. Over the past decades, phage λ has been extensively studied, especially its interaction with the Escherichia coli LamB (EcLamB) protein receptor. Nonetheless, despite the enormous numbers and near-ubiquity of environmental phages, aside from phage λ, there is a paucity of information on other phages which target EcLamB as a receptor. In this study, to answer the question of whether there are other EcLamB-targeting phages in the natural environment, a simple and convenient method was developed and used for isolating environmental phages which target a particular surface structure of a particular bacterium; in this case, the EcLamB outer membrane protein. From the enrichments with the engineered bacterial hosts, a collection of EcLamB-targeting phages (ΦZZ phages) were easily isolated. Intriguingly, unlike phage λ, an obligate EcLamB-dependent phage in the Siphoviridae family, the newly isolated ΦZZ phages alternatively recognised EcLamB or E. coli OmpC (EcOmpC) as a receptor when infecting E. coli. Furthermore, ΦZZ phages were suggested to represent new species in the Tequatrovirus genus in the Myoviridae family, based on phage morphology and genomic sequences. Most phages are thought to have a narrow host range due to their exquisite specificity in receptor recognition. With the ability to optionally recognise two receptors, ΦZZ phages were considered relatively promiscuous. Via the heterologous expression of EcLamB on the bacterial cell surface, the host range of ΦZZ phages was further extended to three different enterobacterial genera. Besides, an interesting selection of evolved phage mutants with a broader host range was isolated, and the key mutations involved in their evolution to adapt to new hosts were investigated by genomic analysis. Finally, and importantly, two ΦZZ phages were found to be putative generalised transducers, which could be exploited as tools for DNA manipulations.

Keywords: environmental microbiology, phage, microbe-host interactions, microbial ecology

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5443 The Significance of Ernest Hemingway's Writing Style in the Development of Georgian Prose of 1950-1960s

Authors: Natia Kvachakidze

Abstract:

The given research aims to study and analyze the influence of Ernest Hemingway’s writing style on Georgian prose of 1950s and 1960s. It is universally known that Ernest Hemingway’s unique writing style has had an enormous effect on various writers. His work remains highly relevant and influential even today. This is especially true about the works written in English, but literary prose created in other languages is not an exception. Certain stylistic peculiarities characteristic for Hemingway’s writing can be traced in literary works written in various languages. It is particularly interesting for us, Georgians, how all these aspects were reflected in Georgian prose of the second-half of XX century. This particular paper (which is a part of a larger research) focuses on major significant peculiarities of Georgian prose of 1950-1960s that might be connected to Hemingway's writing. In this respect, GuramRcheulishvili’s (1934-1960) works should be particularly distinguished (especially his short fiction), but literary works of other Georgian authors are not at all less important. The research involves the analysis of the prose works of some Georgian writers of the given period in the context of tracing similarities and parallels between them and the characteristic features of Ernest Hemingway’s writing style. The use of everyday language as well as short and simple sentences, a concise and sparse style, repetitions, intense dialogues are some of the essential traits in question. Themes like birth and death, war and violence, family, nature, disillusionment also prove to be vitally important for this research. Complex interconnections between the author, the narrator, and the protagonist (often autobiographical) provide another interesting subject to study. At the same time, this paper aims at studying and revealing how Hemingway’s method was reflected and transformed in Georgian prose. In this respect, it is interesting to trace not only the direct effect of Hemingway’s style but also the role of certain Georgian translations of the works of this American writer.

Keywords: hemingway, prose, georgian writers, writing style

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5442 Remediation Activities in Bagnoli Superfund Site: An Italian Case of Study

Authors: S. Bellagamba, S. Malinconico, P. De Simone, F. Paglietti

Abstract:

Until the 1990s, Italy was among the world’s leading producers of raw asbestos fibres and Asbestos Containing Materials (ACM) and one of the most contaminated Countries in Europe. To reduce asbestos-related health effects, Italy has adopted many laws and regulations regarding exposure thresholds, limits, and remediation tools. The Italian Environmental Ministry (MASE) has identified 42 Italian Superfund sites, 11 of which are mainly contaminated by Asbestos. The highest levels of exposure occur during remediation activities in the 42 superfund-sites and during the management of asbestos containing waste in landfills, which requires specific procedures. INAIL-DIT play a role as MASE scientific consultant on issues concerning pollution, remediation, and Asbestos Containing Waste (ACW) management. The aim is to identify the best Emergency Safety Measures, to suggest specific best pratics for remediation through occupational on site monitorings and laboratory analysis. Moreover, the aim of INAIL research is testing the available technologies for working activities and analytical methodologies. This paper describes the remediation of Bagnoli industrial facility (Naples), an Eternit factory which produced asbestos cement products. The remediation has been analyzed, considering a first phase focused on the demolition of structures and plants and a second phase regarding the characterization, screening, removal, and disposal of polluted soils. The project planned the complete removal of all the asbestos dispersed in the soil and subsoil and the recovery of the clean fraction. This work highlights the remediation techniques used and the prevention measures provide for workers and daily life areas protection. This study, considering the high number of asbestos cement factories in the world, can to serve as an important reference for similar situation at European or international scale.

Keywords: safety, asbestos, workers, contaminated sites, hazardous waste

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5441 (Re)Framing the Muslim Subject: Studying the Artistic Representation of Guantanamo and Abu Ghraib Detainees

Authors: Iqra Raza

Abstract:

This paper attempts to conceptualize the (de)humanization of the Muslim subject in Karen J. Greenberg and Janet Hamlin’s transmedia Sketching Guantanamo through a close study of the aesthetics and semiotics of the text. The Muslim experience, the paper shall argue, is mediated through a (de)humanization confined and incarcerated within the chains of artistic representation. Hamlin’s reliance on the distortions offered by stereotypes is reminiscent of the late Victorian epistemology on criminality, as evidenced most starkly in the sketch of Khalid Sheikh Mohammad. The position of the white artist thus becomes suspect in the enterprise of neo-Victorian ethnography. The visual stories of movement from within Guantanamo become potent; the paper shall argue, especially in juxtaposition with the images of stillness that came out from the detention centers, which portrayed the enactment of violence on individual bodies with a deliberate erasure of faces. So, while art becomes a way for reclaiming subjectivity or humanizing these identifiable bodies, the medium predicates itself on their objectification. The paper shall explore various questions about what it means for the (criminal?) subjects to be rendered into art rather than being photographed. Does art entail a necessary departure from the assumed objectivity of the photographic images? What makes art the preferred medium for (de)humanization of the violated Muslim bodies? What happens when art is produced without a recognition of the ‘precariousness’ of the life being portrayed? Rendering the detainees into art becomes a slippery task complicated by Hamlin’s privileged position outside the glass walls of the court. The paper shall adjourn analysis at the many dichotomies that exist in the text viz. between the White men and the brown, the Muslims and the Christians, Occident and the Orient problematized by Hamlin’s politics, that of a ‘neutral outsider’ which quickly turns on its head and becomes complicity in her deliberate erasure of the violence that shaped and still shapes Guantanamo.

Keywords: Abu Ghraib, Derrida, Guantanamo, graphic journalism, Muslimness, orient, spectrality

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5440 A Systematic Review on Measuring the Physical Activity Level and Pattern in Persons with Chronic Fatigue Syndrome

Authors: Kuni Vergauwen, Ivan P. J. Huijnen, Astrid Depuydt, Jasmine Van Regenmortel, Mira Meeus

Abstract:

A lower activity level and imbalanced activity pattern are frequently observed in persons with chronic fatigue syndrome (CFS) / myalgic encephalomyelitis (ME) due to debilitating fatigue and post-exertional malaise (PEM). Identification of measurement instruments to evaluate the activity level and pattern is therefore important. The objective is to identify measurement instruments suited to evaluate the activity level and/or pattern in patients with CFS/ME and review their psychometric properties. A systematic literature search was performed in the electronic databases PubMed and Web of Science until 12 October 2016. Articles including relevant measurement instruments were identified and included for further analysis. The psychometric properties of relevant measurement instruments were extracted from the included articles and rated based on the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN) checklist. The review was performed and reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. A total of 49 articles and 15 unique measurement instruments were found, but only three instruments were evaluated in patients with CFS/ME: the Chronic Fatigue Syndrome-Activity Questionnaire (CFS-AQ), Activity Pattern Interview (API) and International Physical Activity Questionnaire-Short Form (IPAQ-SF), three self-report instruments measuring the physical activity level. The IPAQ-SF, CFS-AQ and API are all equally capable of evaluating the physical activity level, but none of the three measurement instruments are optimal to use. No studies about the psychometric properties of activity monitors in patients with CFS/ME were found, although they are often used as the gold standard to measure the physical activity pattern. More research is needed to evaluate the psychometric properties of existing instruments, including the use of activity monitors.

Keywords: chronic fatigue syndrome, data collection, physical activity, psychometrics

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5439 Human Leukocyte Antigen Class 1 Phenotype Distribution and Analysis in Persons from Central Uganda with Active Tuberculosis and Latent Mycobacterium tuberculosis Infection

Authors: Helen K. Buteme, Rebecca Axelsson-Robertson, Moses L. Joloba, Henry W. Boom, Gunilla Kallenius, Markus Maeurer

Abstract:

Background: The Ugandan population is heavily affected by infectious diseases and Human leukocyte antigen (HLA) diversity plays a crucial role in the host-pathogen interaction and affects the rates of disease acquisition and outcome. The identification of HLA class 1 alleles and determining which alleles are associated with tuberculosis (TB) outcomes would help in screening individuals in TB endemic areas for susceptibility to TB and to predict resistance or progression to TB which would inevitably lead to better clinical management of TB. Aims: To be able to determine the HLA class 1 phenotype distribution in a Ugandan TB cohort and to establish the relationship between these phenotypes and active and latent TB. Methods: Blood samples were drawn from 32 HIV negative individuals with active TB and 45 HIV negative individuals with latent MTB infection. DNA was extracted from the blood samples and the DNA samples HLA typed by the polymerase chain reaction-sequence specific primer method. The allelic frequencies were determined by direct count. Results: HLA-A*02, A*01, A*74, A*30, B*15, B*58, C*07, C*03 and C*04 were the dominant phenotypes in this Ugandan cohort. There were differences in the distribution of HLA types between the individuals with active TB and the individuals with LTBI with only HLA-A*03 allele showing a statistically significant difference (p=0.0136). However, after FDR computation the corresponding q-value is above the expected proportion of false discoveries (q-value 0.2176). Key findings: We identified a number of HLA class I alleles in a population from Central Uganda which will enable us to carry out a functional characterization of CD8+ T-cell mediated immune responses to MTB. Our results also suggest that there may be a positive association between the HLA-A*03 allele and TB implying that individuals with the HLA-A*03 allele are at a higher risk of developing active TB.

Keywords: HLA, phenotype, tuberculosis, Uganda

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5438 Transit-Oriented Development as a Tool for Building Social Capital

Authors: Suneet Jagdev

Abstract:

Rapid urbanization has resulted in informal settlements on the periphery of nearly all big cities in the developing world due to lack of affordable housing options in the city. Residents of these communities have to travel long distances to get to work or search for jobs in these cities, and women, children and elderly people are excluded from urban opportunities. Affordable and safe public transport facilities can help them expand their possibilities. The aim of this research is to identify social capital as another important element of livable cities that can be protected and nurtured through transit-oriented development, as a tool to provide real resources that can help these transit-oriented communities become self-sustainable. Social capital has been referred to the collective value of all social networks and the inclinations that arise from these networks to do things for each other. It is one of the key component responsible to build and maintain democracy. Public spaces, pedestrian amenities and social equity are the other essential part of Transit Oriented Development models that will be analyzed in this research. The data has been collected through the analysis of several case studies, the urban design strategies implemented and their impact on the perception and on the community´s experience, and, finally, how these focused on the social capital. Case studies have been evaluated on several metrics, namely ecological, financial, energy consumption, etc. A questionnaire and other tools were designed to collect data to analyze the research objective and reflect the dimension of social capital. The results of the questionnaire indicated that almost all the participants have a positive attitude towards this dimensions of building a social capital with the aid of transit-oriented development. Statistical data of the identified key motivators against against demographic characteristics have been generated based on the case studies used for the paper. The findings suggested that there is a direct relation between urbanization, transit-oriented developments, and social capital.

Keywords: better opportunities, low-income settlements, social capital, social inclusion, transit oriented development

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5437 Emerging Therapeutic Approach with Dandelion Phytochemicals in Breast Cancer Treatment

Authors: Angel Champion, Sadia Kanwal, Rafat Siddiqui

Abstract:

Harnessing phytochemicals from plant sources presents a novel opportunity to prevent or treat malignant diseases, including breast cancer. Chemotherapy lacks precision in targeting cancerous cells while sparing normal cells, but a phytopharmaceutical approach may offer a solution. Dandelion, a common weed plant, is rich in phytochemicals and provides a safer, more cost-effective alternative with lower toxicity than traditional pharmaceuticals for conditions such as breast cancer. In this study, an in-vitro experiment will be conducted using the ethanol extract of Dandelion on triple-negative MDA-231 breast cancer cell lines. The polyphenolic analysis revealed that the Dandelion extract, particularly from the root and leaf (both cut and sifted), had the most potent antioxidant properties and exhibited the most potent antioxidation activity from the powdered leaf extract. The extract exhibits prospective promising effects for inducing cell proliferation and apoptosis in breast cancer cells, highlighting its potential for targeted therapeutic interventions. Standardizing methods for Dandelion use is crucial for future clinical applications in cancer treatment. Combining plant-derived compounds with cancer nanotechnology holds the potential for effective strategies in battling malignant diseases. Utilizing liposomes as carriers for phytoconstituent anti-cancer agents offers improved solubility, bioavailability, immunoregulatory effects, advancing anticancer immune function, and reducing toxicity. This integrated approach of natural products and nanotechnology has significant potential to revolutionize healthcare globally, especially in underserved communities where herbal medicine is prevalent.

Keywords: apoptosis, antioxidant activity, cancer nanotechnology, phytopharmaceutical

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5436 Optimization for Guide RNA and CRISPR/Cas9 System Nanoparticle Mediated Delivery into Plant Cell for Genome Editing

Authors: Andrey V. Khromov, Antonida V. Makhotenko, Ekaterina A. Snigir, Svetlana S. Makarova, Natalia O. Kalinina, Valentin V. Makarov, Mikhail E. Taliansky

Abstract:

Due to its simplicity, CRISPR/Cas9 has become widely used and capable of inducing mutations in the genes of organisms of various kingdoms. The aim of this work was to develop applications for the efficient modification of DNA coding sequences of phytoene desaturase (PDS), coilin and vacuolar invertase (Solanum tuberosum) genes, and to develop a new nanoparticles carrier efficient technology to deliver the CRISPR/Cas9 system for editing the plant genome. For each of the genes - coilin, PDS and vacuolar invertase, five single RNA guide (sgRNAs) were synthesized. To determine the most suitable nanoplatform, two types of NP platforms were used: magnetic NPs (MNPS) and gold NPs (AuNPs). To test the penetration efficiency, they were functionalized with fluorescent agents - BSA * FITS and GFP, as well as labeled Cy3 small-sized RNA. To measure the efficiency, a fluorescence and confocal microscopy were used. It was shown that the best of these options were AuNP - both in the case of proteins and in the case of RNA. The next step was to check the possibility of delivering components of the CRISPR/Cas9 system to plant cells for editing target genes. AuNPs were functionalized with a ribonucleoprotein complex consisting of Cas9 and corresponding to target genes sgRNAs, and they were biolistically bombarded to axillary buds and apical meristems of potato plants. After the treatment by the best NP carrier, potato meristems were grown to adult plants. DNA isolated from this plants was sent to a preliminary fragment of the analysis to screen out the non-transformed samples, and then to the NGS. The present work was carried out with the financial support from the Russian Science Foundation (grant No. 16-16-04019).

Keywords: biobombardment, coilin, CRISPR/Cas9, nanoparticles, NPs, PDS, sgRNA, vacuolar invertase

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