Search results for: residential differentiation
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1323

Search results for: residential differentiation

393 Applying EzRAD Method for SNPs Discovery in Population Genetics of Freshwater and Marine Fish in the South of Vietnam

Authors: Quyen Vu Dang Ha, Oanh Truong Thi, Thuoc Tran Linh, Kent Carpenter, Thinh Doan Vu, Binh Dang Thuy

Abstract:

Enzyme restriction site associated DNA (EzRAD) has recently emerged as a promising genomic approach for exploring fish genetic diversity on a genome-wide scale. This is a simplified method for genomic genotyping in non-model organisms and applied for SNPs discovery in the population genetics of freshwater and marine fish in the South of Vietnam. The observations of regional-scale differentiation of commercial freshwater fish (smallscale croakers Boesemania microlepis) and marine fish (emperor Lethrinus lentjan) are clarified. Samples were collected along Hau River and coastal area in the south and center Vietnam. 52 DNA samples from Tra Vinh, An Giang Province for Boesemania microlepis and 34 DNA samples of Lethrinus lentjan from Phu Quoc, Nha Trang, Da Nang Province were used to prepare EzRAD libraries from genomic DNA digested with MboI and Sau3AI. A pooled sample of regional EzRAD libraries was sequenced using the HiSeq 2500 Illumina platform. For Boesemania microlepis, the small scale population different from upstream to downstream of Hau river were detected, An Giang population exhibited less genetic diversity (SNPs per individual from 14 to 926), in comparison to Tra Vinh population (from 11 to 2172). For Lethrinus lentjan, the result showed the minor difference between populations in the Northern and the Southern Mekong River. The numbers of contigs and SNPs vary from 1315 to 2455 and from 7122 to 8594, respectively (P ≤ 0.01). The current preliminary study reveals regional scale population disconnection probably reflecting environmental changing. Additional sampling and EzRad libraries need to be implemented for resource management in the Mekong Delta.

Keywords: Boesemania microlepis, EzRAD, Lethrinus lentjan, SNPs

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392 The Results of the Systematic Archaeological Survey of Sistan (Iran)

Authors: Reza Mehrafarin, Nafiseh Mirshekari

Abstract:

The Sistan plain has always been a site for the settlement of various human societies, thanks to its favorable environmental conditions, such as abundant water from the Hirmand River and fertile sedimentary soil. Consequently, there was a need for a systematic archaeological investigation in the area. The survey had multiple objectives, with the most significant ones being the creation of an archaeological map and the identification and documentation of all ancient sites to establish their records and chronology. The survey was carried out in two phases, with each phase covering half of the area. The research method involved fieldwork, with two teams of professional archaeologists conducting a comprehensive survey of each of the 22 areas in Sistan. Once an area was identified, various recording, scientific, and field operations were executed to study the site. In the first phase (2007), an intensive field survey focused on the residential area of Sistan, including its northern and eastern regions. This phase resulted in the identification of 808 sites in eleven selected areas. In the second phase (2009), the desert area of Sistan, or its southern half, was surveyed, leading to the identification of approximately 853 sites. Overall, these surveys resulted in the identification of 1661 sites in Sistan. Among these sites, approximately 899 belong to the Bronze Age (late 4th millennium BCE to early 2nd millennium BCE). Of these sites, around 501 date back to the historical period, while nearly 590 sites pertain to the Islamic period. The archaeological investigations of both phases revealed that Sistan has consistently possessed fertile soil, abundant water, and a skilled workforce, making it capable of becoming Iran's granary and the center of the East once again if these conditions are restored.

Keywords: sistan, field surveys, archaeology, archaeological map

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391 New Test Algorithm to Detect Acute and Chronic HIV Infection Using a 4th Generation Combo Test

Authors: Barun K. De

Abstract:

Acquired immunodeficiency syndrome (AIDS) is caused by two types of human immunodeficiency viruses, collectively designated HIV. HIV infection is spreading globally particularly in developing countries. Before an individual is diagnosed with HIV, the disease goes through different phases. First there is an acute early phase that is followed by an established or chronic phase. Subsequently, there is a latency period after which the individual becomes immunodeficient. It is in the acute phase that an individual is highly infectious due to a high viral load. Presently, HIV diagnosis involves use of tests that do not detect the acute phase infection during which both the viral RNA and p24 antigen are expressed. Instead, these less sensitive tests detect antibodies to viral antigens which are typically sero-converted later in the disease process following acute infection. These antibodies are detected in both asymptomatic HIV-infected individuals as well as AIDS patients. Studies indicate that early diagnosis and treatment of HIV infection can reduce medical costs, improve survival, and reduce spreading of infection to new uninfected partners. Newer 4th generation combination antigen/antibody tests are highly sensitive and specific for detection of acute and established HIV infection (HIV1 and HIV2) enabling immediate linkage to care. The CDC (Center of Disease Control, USA) recently recommended an algorithm involving three different tests to screen and diagnose acute and established infections of HIV-1 and HIV-2 in a general population. Initially a 4th generation combo test detects a viral antigen p24 and specific antibodies against HIV -1 and HIV-2 envelope proteins. If the test is positive it is followed by a second test known as a differentiation assay which detects antibodies against specific HIV-1 and HIV-2 envelope proteins confirming established infection of HIV-1 or HIV-2. However if it is negative then another test is performed that measures viral load confirming an acute HIV-1 infection. Screening results of a Phoenix area population detected 0.3% new HIV infections among which 32.4% were acute cases. Studies in the U.S. indicate that this algorithm effectively reduces HIV infection through immediate treatment and education following diagnosis.

Keywords: new algorithm, HIV, diagnosis, infection

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390 Structural Insulated Panels

Authors: R. Padmini, G. V. Manoj Kumar

Abstract:

Structural insulated panels (SIPs) are a high-performance building system for residential and light commercial construction. The panels consist of an insulating foam core sandwiched between two structural facings, typically oriented strand board (OSB). SIPs are manufactured under factory controlled conditions and can be fabricated to fit nearly any building design. The result is a building system that is extremely strong, energy efficient and cost effective. Building with SIPs will save you time, money and labor. Building with SIPs generally costs about the same as building with wood frame construction when you factor in the labor savings resulting from shorter construction time and less job-site waste. Other savings are realized because smaller heating and cooling systems are required with SIP construction. Structural insulated panels (SIPs) are one of the most airtight and well-insulated building systems available, making them an inherently green product. An airtight SIP building will use less energy to heat and cool, allow for better control over indoor environmental conditions, and reduce construction waste. Green buildings use less energy, reducing carbon dioxide emissions and playing an important role in combating global climate change. Buildings also use a tremendous amount of natural resources to construct and operate. Constructing green buildings that use these resources more efficiently, while minimizing pollution that can harm renewable natural resources, is crucial to a sustainable future.

Keywords: high performance, under factory controlled, wood frame, carbon dioxide emissions, natural resources

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389 Evaluation of Prestressed Reinforced Concrete Slab Punching Shear Using Finite Element Method

Authors: Zhi Zhang, Liling Cao, Seyedbabak Momenzadeh, Lisa Davey

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Reinforced concrete (RC) flat slab-column systems are commonly used in residential or office buildings, as the flat slab provides efficient clearance resulting in more stories at a given height than regular reinforced concrete beam-slab system. Punching shear of slab-column joints is a critical component of two-way reinforced concrete flat slab design. The unbalanced moment at the joint is transferred via slab moment and shear forces. ACI 318 provides an equation to evaluate the punching shear under the design load. It is important to note that the design code considers gravity and environmental load when considering the design load combinations, while it does not consider the effect from differential foundation settlement, which may be a governing load condition for the slab design. This paper describes how prestressed reinforced concrete slab punching shear is evaluated based on ACI 318 provisions and finite element analysis. A prestressed reinforced concrete slab under differential settlements is studied using the finite element modeling methodology. The punching shear check equation is explained. The methodology to extract data for punching shear check from the finite element model is described and correlated with the corresponding code provisions. The study indicates that the finite element analysis results should be carefully reviewed and processed in order to perform accurate punching shear evaluation. Conclusions are made based on the case studies to help engineers understand the punching shear behavior in prestressed and non-prestressed reinforced concrete slabs.

Keywords: differential settlement, finite element model, prestressed reinforced concrete slab, punching shear

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388 Collaborative Stylistic Group Project: A Drama Practical Analysis Application

Authors: Omnia F. Elkommos

Abstract:

In the course of teaching stylistics to undergraduate students of the Department of English Language and Literature, Faculty of Arts and Humanities, the linguistic tool kit of theories comes in handy and useful for the better understanding of the different literary genres: Poetry, drama, and short stories. In the present paper, a model of teaching of stylistics is compiled and suggested. It is a collaborative group project technique for use in the undergraduate diverse specialisms (Literature, Linguistics and Translation tracks) class. Students initially are introduced to the different linguistic tools and theories suitable for each literary genre. The second step is to apply these linguistic tools to texts. Students are required to watch videos performing the poems or play, for example, and search the net for interpretations of the texts by other authorities. They should be using a template (prepared by the researcher) that has guided questions leading students along in their analysis. Finally, a practical analysis would be written up using the practical analysis essay template (also prepared by the researcher). As per collaborative learning, all the steps include activities that are student-centered addressing differentiation and considering their three different specialisms. In the process of selecting the proper tools, the actual application and analysis discussion, students are given tasks that request their collaboration. They also work in small groups and the groups collaborate in seminars and group discussions. At the end of the course/module, students present their work also collaboratively and reflect and comment on their learning experience. The module/course uses a drama play that lends itself to the task: ‘The Bond’ by Amy Lowell and Robert Frost. The project results in an interpretation of its theme, characterization and plot. The linguistic tools are drawn from pragmatics, and discourse analysis among others.

Keywords: applied linguistic theories, collaborative learning, cooperative principle, discourse analysis, drama analysis, group project, online acting performance, pragmatics, speech act theory, stylistics, technology enhanced learning

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387 Recyclable Household Solid Waste Generation and Collection in Beijing, China

Authors: Tingting Liu, Yufeng Wu, Xi Tian, Yu Gong, Tieyong Zuo

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The household solid waste generated by household in Beijing is increasing quickly due to rapid population growth and lifestyle changes. However, there are no rigorous data on the generation and collection of the recyclable household solid wastes. The Beijing city government needs this information to make appropriate policies and plans for waste management. To address this information need, we undertook the first comprehensive study of recyclable household solid waste for Beijing. We carried out a survey of 500 families across sixteen districts in Beijing. We also analyzed the quantities, spatial distribution and categories of collected waste handled by curbside recyclers and permanent recycling centers for 340 of the 9797 city-defined residential areas of Beijing. From our results, we estimate that the total quantity of recyclable household solid waste was 1.8 million tonnes generated by Beijing household in 2013 and 71.6% of that was collected. The main generation categories were waste paper (24.4%), waste glass bottle (23.7%) and waste furniture (14.3%). The recycling rate was varied among different kinds of municipal solid waste. Also based on our study, we estimate there were 22.8 thousand curbside recyclers and 5.7 thousand permanent recycling centers in Beijing. The problems of household solid waste collecting system were inadequacies of authorized collection centers, skewed ratios of curbside recyclers and authorized permanent recycling centers, weak recycling awareness of residents and lack of recycling resources statistics and appraisal system. According to the existing problems, we put forward the suggestions to improve household solid waste management.

Keywords: Municipal waste; Recyclable waste; Waste categories; Waste collection

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386 The Pricing-Out Phenomenon in the U.S. Housing Market

Authors: Francesco Berald, Yunhui Zhao

Abstract:

The COVID-19 pandemic further extended the multi-year housing boom in advanced economies and emerging markets alike against massive monetary easing during the pandemic. In this paper, we analyze the pricing-out phenomenon in the U.S. residential housing market due to higher house prices associated with monetary easing. We first set up a stylized general equilibrium model and show that although monetary easing decreases the mortgage payment burden, it would raise house prices and lower housing affordability for first-time homebuyers (through the initial housing wealth channel and the liquidity constraint channel that increases repeat buyers’ housing demand), and increase housing wealth inequality between first-time and repeat homebuyers. We then use the U.S. household-level data to quantify the effect of the house price change on housing affordability relative to that of the interest rate change. We find evidence of the pricing-out effect for all homebuyers; moreover, we find that the pricing-out effect is stronger for first-time homebuyers than for repeat homebuyers. The paper highlights the importance of accounting for general equilibrium effects and distributional implications of monetary policy while assessing housing affordability. It also calls for complementing monetary easing with well-targeted policy measures that can boost housing affordability, particularly for first-time and lower-income households. Such measures are also needed during aggressive monetary tightening, given that the fall in house prices may be insufficient or too slow to fully offset the immediate adverse impact of higher rates on housing affordability.

Keywords: pricing-out, U.S. housing market, housing affordability, distributional effects, monetary policy

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385 An Approach towards Designing an Energy Efficient Building through Embodied Energy Assessment: A Case of Apartment Building in Composite Climate

Authors: Ambalika Ekka

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In today’s world, the growing demand for urban built forms has resulted in the production and consumption of building materials i.e. embodied energy in building construction, leading to pollution and greenhouse gas (GHG) emissions. Therefore, new buildings will offer a unique opportunity to implement more energy efficient building without compromising on building performance of the building. Embodied energy of building materials forms major contribution to embodied energy in buildings. The paper results in an approach towards designing an energy efficient apartment building through embodied energy assessment. This paper discusses the trend of residential development in Rourkela, which includes three case studies of the contemporary houses, followed by architectural elements, number of storeys, predominant material use and plot sizes using primary data. It results in identification of predominant material used and other characteristics in urban area. Further, the embodied energy coefficients of various dominant building materials and alternative materials manufactured in Indian Industry is taken in consideration from secondary source i.e. literature study. The paper analyses the embodied energy by estimating materials and operational energy of proposed building followed by altering the specifications of the materials based on the building components i.e. walls, flooring, windows, insulation and roof through res build India software and comparison of different options is assessed with consideration of sustainable parameters. This paper results that autoclaved aerated concrete block only reaches the energy performance Index benchmark i.e. 69.35 kWh/m2 yr i.e. by saving 4% of operational energy and as embodied energy has no particular index, out of all materials it has the highest EE 23206202.43  MJ.

Keywords: energy efficient, embodied energy, EPI, building materials

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384 Study on the Spatial Vitality of Waterfront Rail Transit Station Area: A Case Study of Main Urban Area in Chongqing

Authors: Lianxue Shi

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Urban waterfront rail transit stations exert a dual impact on both the waterfront and the transit station, resulting in a concentration of development elements in the surrounding space. In order to more effectively develop the space around the station, this study focuses on the perspective of the integration of station, city, and people. Taking Chongqing as an example, based on the Arc GIS platform, it explores the vitality of the site from the three dimensions of crowd activity heat, space facilities heat, and spatial accessibility. It conducts a comprehensive evaluation and interpretation of the vitality surrounding the waterfront rail transit station area in Chongqing. The study found that (1) the spatial vitality in the vicinity of waterfront rail transit stations is correlated with the waterfront's functional zoning and the intensity of development. Stations situated in waterfront residential and public spaces are more likely to experience a convergence of people, whereas those located in waterfront industrial areas exhibit lower levels of vitality. (2) Effective transportation accessibility plays a pivotal role in maintaining a steady flow of passengers and facilitating their movement. However, the three-dimensionality of urban space in mountainous regions is a notable challenge, leading to some stations experiencing limited accessibility. This underscores the importance of enhancing the optimization of walking space, particularly the access routes from the station to the waterfront area. (3) The density of spatial facilities around waterfront stations in old urban areas lags behind the population's needs, indicating a need to strengthen the allocation of relevant land and resources in these areas.

Keywords: rail transit station, waterfront, influence area, spatial vitality, urban vitality

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383 The Assessment of Particulate Matter Pollution in Kaunas Districts

Authors: Audrius Dedele, Aukse Miskinyte

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Air pollution is a major problem, especially in large cities, causing a variety of environmental issues and a risk to human health effects. In order to observe air quality, to reduce and control air pollution in the city, municipalities are responsible for the creation of air quality management plans, air quality monitoring and emission inventories. Atmospheric dispersion modelling systems, along with monitoring, are powerful tools, which can be used not only for air quality management, but for the assessment of human exposure to air pollution. These models are widely used in epidemiological studies, which try to determine the associations between exposure to air pollution and the adverse health effects. The purpose of this study was to determine the concentration of particulate matter smaller than 10 μm (PM10) in different districts of Kaunas city during winter season. ADMS-Urban dispersion model was used for the simulation of PM10 pollution. The inputs of the model were the characteristics of stationary, traffic and domestic sources, emission data, meteorology and background concentrations were entered in the model. To assess the modelled concentrations of PM10 in Kaunas districts, geographic information system (GIS) was used. More detailed analysis was made using Spatial Analyst tools. The modelling results showed that the average concentration of PM10 during winter season in Kaunas city was 24.8 µg/m3. The highest PM10 levels were determined in Zaliakalnis and Aleksotas districts with are the highest number of individual residential properties, 32.0±5.2 and 28.7±8.2 µg/m3, respectively. The lowest pollution of PM10 was modelled in Petrasiunai district (18.4 µg/m3), which is characterized as commercial and industrial neighbourhood.

Keywords: air pollution, dispersion model, GIS, Particulate matter

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382 Triplex Detection of Pistacia vera, Arachis hypogaea and Pisum sativum in Processed Food Products Using Probe Based PCR

Authors: Ergün Şakalar, Şeyma Özçirak Ergün, Emrah Yalazi̇, Emine Altinkaya, Cengiz Ataşoğlu

Abstract:

In recent years, food allergies which cause serious health problems affect to public health around the world. Foodstuffs which contain allergens are either intentionally used as ingredients or are encased as contaminant in food products. The prevalence of clinical allergy to peanuts and nuts is estimated at about 0.4%-1.1% of the adult population, representing the allergy to pistachio the 7% of the cases of tree nut causing allergic reactions. In order to protect public health and enforce the legislation, methods for sensitive analysis of pistachio and peanut contents in food are required. Pea, pistachio and peanut are used together, to reduce the cost in food production such as baklava, snack foods.DNA technology-based methods in food analysis are well-established and well-roundedtools for species differentiation, allergen detection. Especially, the probe-based TaqMan real-time PCR assay can amplify target DNA with efficiency, specificity, and sensitivity.In this study, pistachio, peanut and pea were finely ground and three separate series of triplet mixtures containing 0.1, 1, 10, 100, 1000, 10,000 and 100,000 mg kg-1 of each sample were prepared for each series, to a final weight of 100 g. DNA from reference samples and industrial products was successfully extracted with the GIDAGEN® Multi-Fast DNA Isolation Kit. TaqMan probes were designed for triplex determination of ITS, Ara h 3 and pea lectin genes which are specific regions for identification pistachio, peanut and pea, respectively.The real-time PCR as quantitative detected pistachio, peanut and pea in these mixtures down to the lowest investigated level of 0.1, 0.1 and 1 mg kg-1, respectively. Also, the methods reported here are capable of detecting of as little as 0.001% level of peanut DNA, 0,000001% level of pistachio DNA and 0.000001% level of pea DNA. We accomplish that the quantitative triplex real-time PCR method developed in this study canbe applied to detect pistachio, peanut and peatraces for three allergens at once in commercial food products.

Keywords: allergens, DNA, real-time PCR, TaqMan probe

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381 Intersectional Perspectives on Gender Equality in Higher Education: A Survey on Swiss Universities of Applied Science

Authors: Birgit Schmid, Brigitte Liebig, Susanne Burren, Maritza Le Breton, Martin Boehnel, Celestina Porta

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Internationalization of students is part of the agenda of many universities worldwide. Yet, how well do universities achieve to guarantee educational success for male and female students of migrant background? This contribution aims on analyzing the effects of the Swiss university environment on perceived educational outcome of migrant students from a gender sensitive perspective. Social selectivity and gender inequalities strongly influence students’ access and success at universities. However, the complex interaction between universities and their disciplinary environments, and educational success of migrant students of both sex remains rarely examined so far. Starting from an intersectional perspective and neo-institutional approaches on higher education organizations, this contribution addresses formal/informal factors in the university environment in its impact on male/female students’ perception of well-being, success and dropout motivation. The paper starts from a most recent Swiss online-survey of Bachelor-students in two Universities of Applied Science and a University of Education in Switzerland. It compares students’ perspectives in four large BA degree courses with different male/female ratio, i.e. educational science, technical/computer science, economy, and social work (N=9`608). Results highlight the complex interplay of gender, migrant background and further dimensions of social differentiation on students’ perception in these different fields of education. Further, they illustrate correlations between students’ perception of discriminatory contexts, poor ratings of social integration and study success, as well a higher rate of dropout ideas. The paper lines out, that formal aspects of internationalization are less important for successfully integrating male/female migrant students than informal university conditions, such as a culture of diversity, which has to become integral part of internationalization strategies.

Keywords: gender and migration, higher education, internationalization, success

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380 Modeling of Cf-252 and PuBe Neutron Sources by Monte Carlo Method in Order to Develop Innovative BNCT Therapy

Authors: Marta Błażkiewicz, Adam Konefał

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Currently, boron-neutron therapy is carried out mainly with the use of a neutron beam generated in research nuclear reactors. This fact limits the possibility of realization of a BNCT in centers distant from the above-mentioned reactors. Moreover, the number of active nuclear reactors in operation in the world is decreasing due to the limited lifetime of their operation and the lack of new installations. Therefore, the possibilities of carrying out boron-neutron therapy based on the neutron beam from the experimental reactor are shrinking. However, the use of nuclear power reactors for BNCT purposes is impossible due to the infrastructure not intended for radiotherapy. Therefore, a serious challenge is to find ways to perform boron-neutron therapy based on neutrons generated outside the research nuclear reactor. This work meets this challenge. Its goal is to develop a BNCT technique based on commonly available neutron sources such as Cf-252 and PuBe, which will enable the above-mentioned therapy in medical centers unrelated to nuclear research reactors. Advances in the field of neutron source fabrication make it possible to achieve strong neutron fluxes. The current stage of research focuses on the development of virtual models of the above-mentioned sources using the Monte Carlo simulation method. In this study, the GEANT4 tool was used, including the model for simulating neutron-matter interactions - High Precision Neutron. Models of neutron sources were developed on the basis of experimental verification based on the activation detectors method with the use of indium foil and the cadmium differentiation method allowing to separate the indium activation contribution from thermal and resonance neutrons. Due to the large number of factors affecting the result of the verification experiment, the 10% discrepancy between the simulation and experiment results was accepted.

Keywords: BNCT, virtual models, neutron sources, monte carlo, GEANT4, neutron activation detectors, gamma spectroscopy

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379 Effect of Diazepam on Internal Organs of Chrysomya megacephala Using Micro-Computed Tomograph

Authors: Sangkhao M., Butcher B. A.

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Diazepam (known as valium) is a medication for calming effect. Many reports on committed suicide cases shown that diazepam is frequently used for this purpose. This research aims to study effect of diazepam on the development of forensically important blowflies, Chrysomya megacephala (Diptera: Calliphoridae) using micro-computed tomography (micro CT). In this study, four rabbits were treated with three different lethal doses of diazepam and one control (LD₀, LD₅₀, LD₁₀₀ and LC). The rabbit’s livers were removed for rearing the blowflies. Pupae were sampled for two series (ages; S1: 24h and S2: 120h) of development. After preparing the specimens, all samples were performed Micro CT using Skyscan 1172. The results shown the effect of diazepam on internal organs and tissues such as brain, cavity of the body, gas bubble, meconium and especially fat body. In the control group, in series 1 (LCS1), fat body was equally dispersed in the head, thorax, and abdomen, development of internal organs were not completed, however, brain, thoracic muscle, wings, legs and rectum were able to observe at 24h after developing into the pupal stage. Development of each organ in the control group in the series two was completed. In the treatment groups, LD₀, LD₅₀, LD₁₀₀ (Series 1 and Series 2), tissues are different, such as gas bubble in LD₀S1, was observed due to rapidity morphological changes during the metamorphosis of blowfly’s pupa in this treatment. Meconium was observed in LD₅₀S2 group because excretion of metabolic waste was not completed. All of the samples in the treatment groups had differentiation of fat bodies because metabolic activities were not completed and these changes affected on functions of every internal system. Discovering of differentiated fat bodies are important results because fat bodies of insect functions as liver in human, therefore it is shown that toxin eliminates from blowfly’s body and homeostatic maintenance of the hemolymph proteins, lipid and carbohydrates in each treatment group are abnormal.

Keywords: forensic toxicology, forensic entomology, diptera, diazepam

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378 Impact of a Home-Based Health Intervention on Older Adults at Risk of Chronic Diseases: A Study Protocol

Authors: Elaine Wong Yee-Sing

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Older adults are at high risk of chronic health conditions in Singapore. A closer examination at all facets of their aging process has revealed that they may not be necessary aging well. This demands for an increasing healthcare services brought to their home environment due to limited mobility and in the interest of time management. The home environment is an ideal setting to implement self-directed health promoting activities at their convenience and enable family’s support and motivation. This research protocol aims to explore their healthcare concerns, and creation of age appropriate interventions targeted to improve their chronic disease biomarkers. Convenience sampling of 130 families residing in private housing within five major districts in Singapore will be selected to participate in the health intervention. Statistical Package for Social Science 25 will be used to examine the pre and post screening results of their lipid, glycaemia and anthropometric outcomes. Using focus interviews, data results will be translated and transcribed to investigate on enablers, barriers and improvement on these services. Both qualitative and quantitative research outcomes are crucial to examine the impact of these services for these older adults living in private housing as they are not exposed to government subsidized community health programs. It is hypothesized that provision of relevant yet engaging health programs at their homes may mitigate the rising burden of chronic health conditions and result in successful aging outcomes among older Singaporeans.

Keywords: chronic diseases, health program, older adults, residential homes

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377 Analyzing the Effect of Materials’ Selection on Energy Saving and Carbon Footprint: A Case Study Simulation of Concrete Structure Building

Authors: M. Kouhirostamkolaei, M. Kouhirostami, M. Sam, J. Woo, A. T. Asutosh, J. Li, C. Kibert

Abstract:

Construction is one of the most energy consumed activities in the urban environment that results in a significant amount of greenhouse gas emissions around the world. Thus, the impact of the construction industry on global warming is undeniable. Thus, reducing building energy consumption and mitigating carbon production can slow the rate of global warming. The purpose of this study is to determine the amount of energy consumption and carbon dioxide production during the operation phase and the impact of using new shells on energy saving and carbon footprint. Therefore, a residential building with a re-enforced concrete structure is selected in Babolsar, Iran. DesignBuilder software has been used for one year of building operation to calculate the amount of carbon dioxide production and energy consumption in the operation phase of the building. The primary results show the building use 61750 kWh of energy each year. Computer simulation analyzes the effect of changing building shells -using XPS polystyrene and new electrochromic windows- as well as changing the type of lighting on energy consumption reduction and subsequent carbon dioxide production. The results show that the amount of energy and carbon production during building operation has been reduced by approximately 70% by applying the proposed changes. The changes reduce CO2e to 11345 kg CO2/yr. The result of this study helps designers and engineers to consider material selection’s process as one of the most important stages of design for improving energy performance of buildings.

Keywords: construction materials, green construction, energy simulation, carbon footprint, energy saving, concrete structure, designbuilder

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376 Urban Design as a Tool to Address Safety in a Crime Ridden Area: A Case Study of Malviya Nagar, New Delhi

Authors: Shramana Mondal

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As a city is growing in population, sprawl, and complexity, use of public spaces increases variably and thus ensuring safety for the people becomes an utmost priority. While active monitoring measures may be necessary in some places, urban design can play a major role in devising self-policing and encourage active public life. This paper aims to explore the various spatial and psychological reasons for the occurrence of crime and the role of ‘urban design’ to address this issue. In this research, the principles of urban design are examined, as well as projected on actual site by addressing the issue with urban design principles. In this review the sociological, psychological, typological and morphological factors are addressed which affect the safety of a space and the possible framing guidelines, controls and urban design strategies are explored to address a safe neighborhood. On the basis of statistical survey, the residential and street network of Malviya Nagar in Delhi is chosen as the area of demonstration. The programs inhibit a safe neighborhood and a movement network that are addressed based on the four principles of natural surveillance, territoriality, community building, and connectivity. The paper concludes with a discussion of the urban design as an effective tool by creating an intense active zone with mixed use feature to ensure throughout activity and also ensuring safe pedestrian zone by introducing sense of community feeling and territoriality thus achieving active, useful and public friendly space.

Keywords: crime, public life, safety, urban design

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375 High-Rises and Urban Design: The Reasons for Unsuccessful Placemaking with Residential High-Rises in England

Authors: E. Kalcheva, A. Taki, Y. Hadi

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High-rises and placemaking is an understudied combination which receives more and more interest with the proliferation of this typology in many British cities. The reason for studying three major cities in England: London, Birmingham and Manchester, is to learn from the latest advances in urban design in well-developed and prominent urban environment. The analysis of several high-rise sites reveals the weaknesses in urban design of contemporary British cities and presents an opportunity to study from the implemented examples. Therefore, the purpose of this research is to analyze design approaches towards creating a sustainable and varied urban environment when high-rises are involved. The research questions raised by the study are: what is the quality of high-rises and their surroundings; what facilities and features are deployed in the research area; what is the role of the high-rise buildings in the placemaking process; what urban design principles are applicable in this context. The methodology utilizes observation of the researched area by structured questions, developed by the author to evaluate the outdoor qualities of the high-rise surroundings. In this context, the paper argues that the quality of the public realm around the high-rises is quite low, missing basic but vital elements such as plazas, public art, and seating, along with landscaping and pocket parks. There is lack of coherence, the rhythm of the streets is often disrupted, and even though the high-rises are very aesthetically appealing, they fail to create a sense of place on their own. The implications of the study are that future planning can take into consideration the critique in this article and provide more opportunities for urban design interventions around high-rise buildings in the British cities.

Keywords: high-rises, placemaking, urban design, townscape

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374 Expression of ULK-1 mRNA in Human Peripheral Blood Mononuclear Cells from Patients with Alzheimer's Disease

Authors: Ali Bayram, Remzi Yiğiter

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Objective: Alzheimer's disease (AD), the most common cause of dementia, is a progressive neurodegenerative disease. At present, diagnosis of AD is rather late in the disease. Therefore, we attempted to find peripheral biomarkers for the early diagnosis of AD. Herein, we conducted a study to investigate the unc-51 like autophagy activating kinase-1 (ULK1) mRNA expression levels in human peripheral blood mononuclear cells from patients with Alzheimer's disease. Method: To determine whether ULK1 gene expression are altered in AD patients, we measured their gene expression in human peripheral blood cell in 50 patients with AD and 50 age and gender matched healthy controls by quantitative real-time PCR technique. Results: We found that both ULK1 gene expression in peripheral blood cell were significantly decreased in patients with AD as compared with controls (p <0.05). Lower levels of ULK1 gene expression were significantly associated with the increased risk for AD. Conclusions: Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2, and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. Alzheimer is the most common neurodegenerative disease. Our results provide useful information that the ULK1 gene expression is decreased in the neurodegeneration and AD patients with, indicating their possible systemic involvement in AD.

Keywords: Alzheimer’s sisease, ULK1, mRNA expression, RT-PCR

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373 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

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372 Predicting Factors for Occurrence of Cardiac Arrest in Critical, Emergency and Urgency Patients in an Emergency Department

Authors: Angkrit Phitchayangkoon, Ar-Aishah Dadeh

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Background: A key aim of triage is to identify the patients with high risk of cardiac arrest because they require intensive monitoring, resuscitation facilities, and early intervention. We aimed to identify the predicting factors such as initial vital signs, serum pH, serum lactate level, initial capillary blood glucose, and Modified Early Warning Score (MEWS) which affect the occurrence of cardiac arrest in an emergency department (ED). Methods: We conducted a retrospective data review of ED patients in an emergency department (ED) from 1 August 2014 to 31 July 2016. Significant variables in univariate analysis were used to create a multivariate analysis. Differentiation of predicting factors between cardiac arrest patient and non-cardiac arrest patients for occurrence of cardiac arrest in an emergency department (ED) was the primary outcome. Results: The data of 527 non-trauma patients with Emergency Severity Index (ESI) 1-3 were collected. The factors found to have a significant association (P < 0.05) in the non-cardiac arrest group versus the cardiac arrest group at the ED were systolic BP (mean [IQR] 135 [114,158] vs 120 [90,140] mmHg), oxygen saturation (mean [IQR] 97 [89,98] vs 82.5 [78,95]%), GCS (mean [IQR] 15 [15,15] vs 11.5 [8.815]), normal sinus rhythm (mean 59.8 vs 30%), sinus tachycardia (mean 46.7 vs 21.7%), pH (mean [IQR] 7.4 [7.3,7.4] vs 7.2 [7,7.3]), serum lactate (mean [IQR] 2 [1.1,4.2] vs 7 [5,10.8]), and MEWS score (mean [IQR] 3 [2,5] vs 5 [3,6]). A multivariate analysis was then performed. After adjusting for multiple factors, ESI level 2 patients were more likely to have cardiac arrest in the ER compared with ESI 1 (odds ratio [OR], 1.66; P < 0.001). Furthermore, ESI 2 patients were more likely than ESI 1 patients to have cardiovascular disease (OR, 1.89; P = 0.01), heart rate < 55 (OR, 6.83; P = 0.18), SBP < 90 (OR, 3.41; P = 0.006), SpO2 < 94 (OR, 4.76; P = 0.012), sinus tachycardia (OR, 4.32; P = 0.002), lactate > 4 (OR, 10.66; P = < 0.001), and MEWS > 4 (OR, 4.86; P = 0.028). These factors remained predictive of cardiac arrest at the ED. Conclusion: The factors related to cardiac arrest in the ED are ESI 1 patients, ESI 2 patients, patients diagnosed with cardiovascular disease, SpO2 < 94, lactate > 4, and a MEWS > 4. These factors can be used as markers in the event of simultaneous arrival of many patients and can help as a pre-state for patients who have a tendency to develop cardiac arrest. The hemodynamic status and vital signs of these patients should be closely monitored. Early detection of potentially critical conditions to prevent critical medical intervention is mandatory.

Keywords: cardiac arrest, predicting factor, emergency department, emergency patient

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371 Analysis of Seismic Waves Generated by Blasting Operations and their Response on Buildings

Authors: S. Ziaran, M. Musil, M. Cekan, O. Chlebo

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The paper analyzes the response of buildings and industrially structures on seismic waves (low frequency mechanical vibration) generated by blasting operations. The principles of seismic analysis can be applied for different kinds of excitation such as: earthquakes, wind, explosions, random excitation from local transportation, periodic excitation from large rotating and/or machines with reciprocating motion, metal forming processes such as forging, shearing and stamping, chemical reactions, construction and earth moving work, and other strong deterministic and random energy sources caused by human activities. The article deals with the response of seismic, low frequency, mechanical vibrations generated by nearby blasting operations on a residential home. The goal was to determine the fundamental natural frequencies of the measured structure; therefore it is important to determine the resonant frequencies to design a suitable modal damping. The article also analyzes the package of seismic waves generated by blasting (Primary waves – P-waves and Secondary waves S-waves) and investigated the transfer regions. For the detection of seismic waves resulting from an explosion, the Fast Fourier Transform (FFT) and modal analysis, in the frequency domain, is used and the signal was acquired and analyzed also in the time domain. In the conclusions the measured results of seismic waves caused by blasting in a nearby quarry and its effect on a nearby structure (house) is analyzed. The response on the house, including the fundamental natural frequency and possible fatigue damage is also assessed.

Keywords: building structure, seismic waves, spectral analysis, structural response

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370 Cars in a Neighborhood: A Case of Sustainable Living in Sector 22 Chandigarh

Authors: Maninder Singh

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The Chandigarh city is under the strain of exponential growth of car density across various neighborhood. The consumerist nature of society today is to be blamed for this menace because everyone wants to own and ride a car. Car manufacturers are busy selling two or more cars per household. The Regional Transport Offices are busy issuing as many licenses to new vehicles as they can in order to generate revenue in the form of Road Tax. The car traffic in the neighborhoods of Chandigarh has reached a tipping point. There needs to be a more empirical and sustainable model of cars per household, which should be based on specific parameters of livable neighborhoods. Sector 22 in Chandigarh is one of the first residential sectors to be established in the city. There is scope to think, reflect, and work out a method to know how many cars we need to sell our citizens before we lose the argument to traffic problems, parking problems, and road rage. This is where the true challenge of a planner or a designer of the city lies. Currently, in Chandigarh city, there are no clear visible answers to this problem. The way forward is to look at spatial mapping, planning, and design of car parking units to address the problem, rather than suggesting extreme measures of banning cars (short-term) or promoting plans for citywide transport (very long-term). This is a chance to resolve the problem with a pragmatic approach from a citizen’s perspective, instead of an orthodox development planner’s methodology. Since citizens are at the center of how the problem is to be addressed, acceptable solutions are more likely to emerge from the car and traffic problem as defined by the citizens. Thus, the idea and its implementation would be interesting in comparison to the known academic methodologies. The novel and innovative process would lead to a more acceptable and sustainable approach to the issue of number of car parks in the neighborhood of Chandigarh city.

Keywords: cars, Chandigarh, neighborhood, sustainable living, walkability

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369 Visual Simulation for the Relationship of Urban Fabric

Authors: Ting-Yu Lin, Han-Liang Lin

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This article is about the urban form of visualization by Cityengine. City is composed of different domains, and each domain has its own fabric because of arrangement. For example, a neighborhood unit contains fabrics such as schools, street networks, residential and commercial spaces. Therefore, studying urban morphology can help us understand the urban form in planning process. Streets, plots, and buildings seem as urban fabrics, and they configure urban form. Traditionally, urban morphology usually discussed single parameter, which is building type, ignoring other parameters such as streets and plots. However, urban space is three-dimensional, instead of two-dimensional. People perceive urban space by their visualization. Therefore, using visualization can fill the gap between two dimensions and three dimensions. Hence, the study of urban morphology will strengthen the understanding of whole appearance of a city. Cityengine is a software which can edit, analyze and monitor the data and visualize the result for GIS, a common tool to analyze data and display the map for urban plan and urban design. Cityengine can parameterize the data of streets, plots and building types and visualize the result in three-dimensional way. The research will reappear the real urban form by visualizing. We can know whether the urban form can be parameterized and the parameterized result can match the real urban form. Then, visualizing the result by software in three dimension to analyze the rule of urban form. There will be three stages of the research. It will start with a field survey of Tainan East District in Taiwan to conclude the relationships between urban fabrics of street networks, plots and building types. Second, to visualize the relationship, it will turn the relationship into codes which Cityengine can read. Last, Cityengine will automatically display the result by visualizing.

Keywords: Cityengine, urban fabric, urban morphology, visual simulation

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368 Service Flow in Multilayer Networks: A Method for Evaluating the Layout of Urban Medical Resources

Authors: Guanglin Song

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(Objective) Situated within the context of China's tiered medical treatment system, this study aims to analyze spatial causes of urban healthcare access difficulties from the perspective of the configuration of healthcare facilities. (Methods) A social network analysis approach is employed to construct a healthcare demand and supply flow network between major residential clusters and various tiers of hospitals in the city.(Conclusion) The findings reveal that:1.there exists overall maldistribution and over-concentration of healthcare resources in Study Area, characterized by structural imbalance; 2.the low rate of primary care utilization in Study Area is a key factor contributing to congestion at higher-tier hospitals, as excessive reliance on these institutions by neighboring communities exacerbates the problem; 3.gradual optimization of the healthcare facility layout in Study Area, encompassing holistic, local, and individual institutional levels, can enhance systemic efficiency and resource balance.(Prospects) This research proposes a method for evaluating urban healthcare resource distribution structures based on service flows within hierarchical networks. It offers spatially targeted optimization suggestions for promoting the implementation of the tiered healthcare system and alleviating challenges related to accessibility and congestion in seeking medical care. Provide some new ideas for researchers and healthcare managers in countries, cities, and healthcare management around the world with similar challenges.

Keywords: flow of public services, urban networks, healthcare facilities, spatial planning, urban networks

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367 Hyaluronic Acid - Alginate Hydrogel for the Transdifferentiation of Testis Cells into Erythrocyte and Hepatocyte-like Cells; A Practice Within an Effective Agent Choice

Authors: Leila Rashki Ghaleno, Mohamad Amin Hajari, Leila Montazeri, Abdolhossein Shahverdi, Mojtaba Rezazadeh Valojerdi

Abstract:

Background: Spermatogonia stem cells (SSCs) exhibit pluripotency, enabling them to undergo differentiation into many cell lineages, including neurons, glia, endothelial cells, and hepatocytes when cultured in vitro. Although the specific mechanisms are not yet fully understood, it has been observed that biopolymer agents, such as hyaluronic acid (HA) and alginate (Alg), have the potential to induce transdifferentiation of SSCs. The current work aimed to examine the process of in vitro spermatogenesis and the conversion of mouse testicular cells into hepatocytes and erythrocyte-like cells utilizing the HA-Alg hydrogel. Method: After being extracted from the testes of a 5-day postpartum mouse (5 DPP), the testicular cells were separated into two enzymatic stages and then put into a composite hydrogel containing 0.5% HA and 1% alginate. On days 14 and 28 of culture, the colonies' growth, the cells' viability, and their histology were assessed. Result: Despite observing significant cell proliferation on day 14 and the development of circular-shaped organoids on day 28, it was noted that the organoids generated in the HA-Alg medium tended to maintain their circular morphology on day 28. Notably, the testicular cells underwent transdifferentiation into cell types resembling erythrocytes and hepatocytes. The hepatocyte-like cells exhibited the presence of glycogen and lipid deposits, indicating their hepatocyte-like characteristics. Interestingly, immunostaining analysis revealed the secretion of albumin and the presence of VEGFR on day 14. However, on day 28, albumin expression was not detected, while the expression of Sox9 (a marker for hepatocytes), Vegf, CD34, and C-kit (markers for erythrocytes) showed increased levels in the gene expression evaluation. Conclusion: The present findings indicated that HA-Alg could be a potent and effective agent for the transdifferentiation of testis cells into erythrocyte and hepatocyte-like cells, as recent studies have confirmed the transformation of SSCs into hepatocyte cells during in vitro culture.

Keywords: 3D culture, mouse testicular cell, hyaluronic acid, liver organoids

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366 Adaptor Protein APPL2 Could Be a Therapeutic Target for Improving Hippocampal Neurogenesis and Attenuating Depressant Behaviors and Olfactory Dysfunctions in Chronic Corticosterone-induced Depression

Authors: Jiangang Shen

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Olfactory dysfunction is a common symptom companied by anxiety- and depressive-like behaviors in depressive patients. Chronic stress triggers hormone responses and inhibits the proliferation and differentiation of neural stem cells (NSCs) in the hippocampus and subventricular zone (SVZ)-olfactory bulb (OB), contributing to depressive behaviors and olfactory dysfunction. However, the cellular signaling molecules to regulate chronic stress mediated olfactory dysfunction are largely unclear. Adaptor proteins containing the pleckstrin homology domain, phosphotyrosine binding domain, and leucine zipper motif (APPLs) are multifunctional adaptor proteins. Herein, we tested the hypothesis that APPL2 could inhibit hippocampal neurogenesis by affecting glucocorticoid receptor (GR) signaling, subsequently contributing to depressive and anxiety behaviors as well as olfactory dysfunctions. The major discoveries are included: (1) APPL2 Tg mice had enhanced GR phosphorylation under basic conditions but had no different plasma corticosterone (CORT) level and GR phosphorylation under stress stimulation. (2) APPL2 Tg mice had impaired hippocampal neurogenesis and revealed depressive and anxiety behaviors. (3) GR antagonist RU486 reversed the impaired hippocampal neurogenesis in the APPL2 Tg mice. (4) APPL2 Tg mice displayed higher GR activity and less capacity for neurogenesis at the olfactory system with lesser olfactory sensitivity than WT mice. (5) APPL2 negatively regulates olfactory functions by switching fate commitments of NSCs in adult olfactory bulbs via interaction with Notch1 signaling. Furthermore, baicalin, a natural medicinal compound, was found to be a promising agent targeting APPL2/GR signaling and promoting adult neurogenesis in APPL2 Tg mice and chronic corticosterone-induced depression mouse models. Behavioral tests revealed that baicalin had antidepressant and olfactory-improving effects. Taken together, APPL2 is a critical therapeutic target for antidepressant treatment.

Keywords: APPL2, hippocampal neurogenesis, depressive behaviors and olfactory dysfunction, stress

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365 Expression of Micro-RNA268 in Zinc Deficient Rice

Authors: Sobia Shafqat, Saeed Ahmad Qaisrani

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MicroRNAs play an essential role in the regulation and development of all processes in most eukaryotes because of their prospective part as mediators controlling cell growth and differentiation towards the exact position of RNAs response in plants under biotic and abiotic factors or stressors. In a few cases, Zn is oblivious poisonous for plants due to its heavy metal status. Some other metals are extremely toxic, like Cd, Hg, and Pb, but these elements require in rice for the programming of genes under abiotic stress resembling Zn stress when micro RNAs268 was importantly introduced in rice. The micro RNAs overexpressed in transgenic plants with an accumulation of a large amount of melanin dialdehyde, hydrogen peroxide, and an excessive quantity of Zn in the seedlings stage. Let out results for rice pliability under Zn stress micro RNAs act as negative controllers. But the role of micro RNA268 act as a modulator in different ecological condition. It has been explained clearly with a long understanding of the role of micro RNA268 under stress conditions; pliability and practically showed outcome to increase plant sufferance under Zn stress because micro RNAs is an intervention technique for gene regulation in gene expression. The proposed study was experimented with by using genetic factors of Zn stress and toxicity effect on rice plants done at District Vehari, Pakistan. The trial was performed randomly with three replications in a complete block design (RCBD). These blocks were controlled with different concentrations of genetic factors. By overexpression of micro RNA268 rice, seedling growth was not stopped under Zn deficiency due to the accumulation of a large amount of melanin dialdehyde, hydrogen peroxide, and an excessive quantity of Zn in their seedlings. Results showed that micro RNA268 act as a negative controller under Zn stress. In the end, under stress conditions, micro RNA268 showed the necessary function in the tolerance of rice plants. The directorial work sketch gave out high agronomic applications and yield outcomes in rice with a specific amount of Zn application.

Keywords: micro RNA268, zinc, rice, agronomic approach

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364 A Serum- And Feeder-Free Culture System for the Robust Generation of Human Stem Cell-Derived CD19+ B Cells and Antibody-Secreting Cells

Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

Abstract:

Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

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