Search results for: stem cell fate decision

Authors: Subodh Kumar, Sanjeev Kumar Sharma, Gaurav Kaushik, Pramod Avti, Phulen Sarma, Bikash Medhi, Krishan Lal Khanduja

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Background: It is well known that cigarette smoke is one of the causative factors in various lung diseases especially cancer. Carcinogens and oxidant molecules present in cigarette smoke not only damage the cellular constituents (lipids, proteins, DNA) but may also regulate the molecular pathways involved in inflammation and cancer. Continuous oxidative stress caused by the constituents of cigarette smoke leads to higher PhospholipaseA₂ (PLA₂) activity, resulting in elevated levels of secondary metabolites whose role is well defined in cancer. To reduce the burden of chronic inflammation as well as oxidative stress, and higher levels of secondary metabolites, we checked the curative potential of PLA₂ inhibitor Bromoenol Lactone (BEL) during continuous exposure of cigarette smoke condensate (CSC). Aim: To check the therapeutic potential of Bromoenol Lactone (BEL), an inhibitor of PhospholipaseA₂s, in pathways of CSC-induced changes in type I and type II alveolar epithelial cells. Methods: Effect of BEL on CSC-induced PLA2 activity were checked using colorimetric assay, cellular toxicity using cell viability assay, membrane integrity using fluorescein di-acetate (FDA) uptake assay, reactive oxygen species (ROS) levels and apoptosis markers through flow cytometry, and cellular regulation using MAPKinases levels, in lung epithelium. Results: BEL significantly mimicked CSC-induced PLA₂ activity, ROS levels, apoptosis, and kinases level whereas improved cellular viability and membrane integrity. Conclusions: Current observations revealed that BEL may be a potential therapeutic agent during Cigarette smoke-induced anomalies in lung epithelium.

Keywords: cigarette smoke condensate, phospholipase A₂, oxidative stress, alveolar epithelium, bromoenol lactone

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Authors: Urbánek Jiří J., Krahulec Josef, Urbánek Jiří F., Johanidesová Jitka

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These Computational assistance for the research and modelling of critical infrastructure subjects continuity deal with this paper. It enables us the using of prevailing operation system MS Office (SmartArt...) for mathematical models, using DYVELOP (Dynamic Vector Logistics of Processes) method. It serves for crisis situations investigation and modelling within the organizations of critical infrastructure. In the first part of the paper, it will be introduced entities, operators and actors of DYVELOP method. It uses just three operators of Boolean algebra and four types of the entities: the Environments, the Process Systems, the Cases and the Controlling. The Process Systems (PrS) have five “brothers”: Management PrS, Transformation PrS, Logistic PrS, Event PrS and Operation PrS. The Cases have three “sisters”: Process Cell Case, Use Case and Activity Case. They all need for the controlling of their functions special Ctrl actors, except ENV – it can do without Ctrl. Model´s maps are named the Blazons and they are able mathematically - graphically express the relationships among entities, actors and processes. In the second part of this paper, the rich blazons of DYVELOP method will be used for the discovering and modelling of the cycling cases and their phases. The blazons need live PowerPoint presentation for better comprehension of this paper mission. The crisis management of energetic crisis infrastructure organization is obliged to use the cycles for successful coping of crisis situations. Several times cycling of these cases is a necessary condition for the encompassment of the both the emergency event and the mitigation of organization´s damages. Uninterrupted and continuous cycling process bring for crisis management fruitfulness and it is a good indicator and controlling actor of organizational continuity and its sustainable development advanced possibilities. The research reliable rules are derived for the safety and reliable continuity of energetic critical infrastructure organization in the crisis situation.

Keywords: blazons, computational assistance, DYVELOP method, critical infrastructure

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Authors: Shang-Yu Chiang, Yu-Shan Tsai, Shih-Hsien Sung, Chung-Ming Lo

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Mitral regurgitation (MR) is a heart disorder which the mitral valve does not close properly when the heart pumps out blood. MR is the most common form of valvular heart disease in the adult population. The diagnostic echocardiographic finding of MR is straightforward due to the well-known clinical evidence. In the determination of MR severity, quantification of sonographic findings would be useful for clinical decision making. Clinically, the vena contracta is a standard for MR evaluation. Vena contracta is the point in a blood stream where the diameter of the stream is the least, and the velocity is the maximum. The quantification of vena contracta, i.e. the vena contracta width (VCW) at mitral valve, can be a numeric measurement for severity assessment. However, manually delineating the VCW may not accurate enough. The result highly depends on the operator experience. Therefore, this study proposed an automatic method to quantify VCW to evaluate MR severity. Based on color Doppler ultrasound, VCW can be observed from the blood flows to the probe as the appearance of red or yellow area. The corresponding brightness represents the value of the flow rate. In the experiment, colors were firstly transformed into HSV (hue, saturation and value) to be closely align with the way human vision perceives red and yellow. Using ellipse to fit the high flow rate area in left atrium, the angle between the mitral valve and the ultrasound probe was calculated to get the vertical shortest diameter as the VCW. Taking the manual measurement as the standard, the method achieved only 0.02 (0.38 vs. 0.36) to 0.03 (0.42 vs. 0.45) cm differences. The result showed that the proposed automatic VCW extraction can be efficient and accurate for clinical use. The process also has the potential to reduce intra- or inter-observer variability at measuring subtle distances.

Keywords: mitral regurgitation, vena contracta, color doppler, image processing

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Authors: Monika Michałowska, Anna Alichniewicz

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Detrimental outcomes of advanced maternal age on the chances of fertilization, pregnancy as well as mother and fetus health have been recognized for several decades. It seemed interesting to investigate whether there is a comparable awareness of the detrimental influence on the reproductive outcomes of late fatherhood, given that it has been already ten years since an intense and growing interest concerning later-age fatherhood commenced in medical research. To address that issue a two-step research was done. First, we performed a review of the subject literature to answer the following questions: 1) What age is defined as advanced?; 2) Is the same age defined as advanced in both genders?; 3) What terminology concerning age issues is used?; 4) Is the same age terminology used regarding both genders? The second part of our studies was devoted to the views of medical students. This part of our research comprised both quantitative and qualitative studies. Opinions of medical students in one of the Polish medical universities on several issues connected with assisted reproduction technology (ART) were gathered: 1) students’ attitude to in vitro fertilization (IVF) for women over 40 and for postmenopausal women; 2) students’ attitude to late fatherhood; 3) students’ reasoning given against acceptability of IVF procedure for all of these group of patients involved in an IVF procedure. Our analyses revealed that: First, there is no universal definition of the term ‘advanced age’; secondly, there is a general tendency to adopt different age limits depending on whether they refer to maternal or paternal age, but no justification is provided by the researchers explaining why they set different age limits for women and men; thirdly, the image of postponed fatherhood stands in stark contrast to postponed motherhood - while postponed fatherhood is frequently portrayed as a reasonable and conscious decision enabling a stable family environment for a child, the reasonableness of postponed motherhood is often questioned; finally, the bias regarding maternal versus paternal age is deeply embedded in medical students’ attitude to IVF for women over 40 and for postmenopausal women.

Keywords: gender stereotypes, reproductive medicine, maternal age, paternal age

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Authors: Xiaowei Zhang, Min Xu, Saeid Habibi, Fengjun Yan, Ryan Ahmed

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Recently, Electric Vehicles (EVs) have received extensive consideration since they offer a more sustainable and greener transportation alternative compared to fossil-fuel propelled vehicles. Lithium-Ion (Li-ion) batteries are increasingly being deployed in EVs because of their high energy density, high cell-level voltage, and low rate of self-discharge. Since Li-ion batteries represent the most expensive component in the EV powertrain, accurate monitoring and control strategies must be executed to ensure their prolonged lifespan. The Battery Management System (BMS) has to accurately estimate parameters such as the battery State-of-Charge (SOC), State-of-Health (SOH), and Remaining Useful Life (RUL). In order for the BMS to estimate these parameters, an accurate and control-oriented battery model has to work collaboratively with a robust state and parameter estimation strategy. Since battery physical parameters, such as the internal resistance and diffusion coefficient change depending on the battery state-of-life (SOL), the BMS has to be adaptive to accommodate for this change. In this paper, an extensive battery aging study has been conducted over 12-months period on 5.4 Ah, 3.7 V Lithium polymer cells. Instead of using fixed charging/discharging aging cycles at fixed C-rate, a set of real-world driving scenarios have been used to age the cells. The test has been interrupted every 5% capacity degradation by a set of reference performance tests to assess the battery degradation and track model parameters. As battery ages, the combined model parameters are optimized and tracked in an offline mode over the entire batteries lifespan. Based on the optimized model, a state and parameter estimation strategy based on the Extended Kalman Filter (EKF) and the relatively new Smooth Variable Structure Filter (SVSF) have been applied to estimate the SOC at various states of life.

Keywords: lithium-ion batteries, genetic algorithm optimization, battery aging test, parameter identification

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Authors: Izabel Cristina Zattar, Gilberto Passos Lima, Guilherme Schünemann de Oliveira

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In Brazil, there are several institutions committed to the development of new businesses based on product innovation. Among them are business incubators, universities and science institutes. Business incubators can be defined as nurseries for new companies, which may be in the technology segment, discussed in this article. Business incubators provide services related to infrastructure, such as physical space and meeting rooms. Besides these services, incubators also offer assistance in the form of information and communication, access to finance, relationship networks and business monitoring and mentoring processes. Business incubators support not all technology companies. One of the business incubators tasks is to assess the nature and feasibility of new business proposals. To assist this goal, this paper proposes a methodology for evaluating new business using the Analytic Hierarchy Process (AHP). This paper presents the concepts used in the assessing methodology application for new business, concepts that have been tested with positive results in practice. This study counts on three main steps: first, a hierarchy was built, based on new business manuals used by the business incubators. These books and manuals relate business selection requirements, such as the innovation status and other technological aspects. Then, a questionnaire was generated, in order to guide incubator experts in the parity comparisons at all hierarchy levels. The weights of each requirement are calculated from information obtained from the questionnaire responses. Finally, the proposed method was applied to evaluate five new business proposals, which were applying to be part of a company incubator. The main result is the classification of these new businesses, which helped the incubator experts to decide what companies were more eligible to work with. This classification may also be helpful to the decision-making process of business incubators in future selection processes.

Keywords: Analytic Hierarchy Process (AHP), Brazilian companies incubators, technology companies, incubator

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Authors: Adel Alblihy, Reem Ali, Mashael Algethami, Ahmed Shoqafi, Michael S. Toss, Juliette Brownlie, Natalie J. Tatum, Ian Hickson, Paloma Ordonez Moran, Anna Grabowska, Jennie N. Jeyapalan, Nigel P. Mongan, Emad A. Rakha, Srinivasan Madhusudan

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Platinum resistance is a clinical challenge in ovarian cancer. Platinating agents induce DNA damage which activate Mre11 nuclease directed DNA damage signalling and response (DDR). Upregulation of DDR may promote chemotherapy resistance. Here we have comprehensively evaluated Mre11 in epithelial ovarian cancers. In clinical cohort that received platinum- based chemotherapy (n=331), Mre11 protein overexpression was associated with aggressive phenotype and poor progression free survival (PFS) (p=0.002). In the ovarian cancer genome atlas (TCGA) cohort (n=498), Mre11 gene amplification was observed in a subset of serous tumours (5%) which correlated highly with Mre11 mRNA levels (p<0.0001). Altered Mre11 levels was linked with genome wide alterations that can influence platinum sensitivity. At the transcriptomic level (n=1259), Mre11 overexpression was associated with poor PFS (p=0.003). ROC analysis showed an area under the curve (AUC) of 0.642 for response to platinum-based chemotherapy. Pre-clinically, Mre11 depletion by gene knock down or blockade by small molecule inhibitor (Mirin) reversed platinum resistance in ovarian cancer cells and in 3D spheroid models. Importantly, Mre11 inhibition was synthetically lethal in platinum sensitive XRCC1 deficient ovarian cancer cells and 3D-spheroids. Selective cytotoxicity was associated with DNA double strand break (DSB) accumulation, S-phase cell cycle arrest and increased apoptosis. We conclude that pharmaceutical development of Mre11 inhibitors is a viable clinical strategy for platinum sensitization and synthetic lethality in ovarian cancer.

Keywords: MRE11; XRCC1, ovarian cancer, platinum sensitization, synthetic lethality

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Authors: Daniel Monsalve, Takafumi Noguchi

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Mycelium-based composites (MBC) are made by growing living mycelium on lignocellulosic fibres to create a porous composite material which can be lightweight, and biodegradable, making them suitable as a sustainable thermal insulation. Thus, they can help to reduce material extraction while improving the energy efficiency of buildings, especially when agricultural by-products are used. However, as MBC are hygroscopic materials, moisture can reduce their thermal insulation efficiency. It is known that surface growth, or “mycelium skin”, can form a natural coating due to the hydrophobic properties in the mycelium cell wall. Therefore, this research aims to biofabricate a homogeneous mycelium skin and measure its influence on the final composite material by testing material properties such as thermal conductivity, vapour permeability and water absorption by partial immersion over 24 hours. In addition, porosity, surface morphology and chemical composition were also analyzed. The white-rot fungi species Pleurotus ostreatus, Ganoderma lucidum, and Trametes versicolor were grown on 10 mm hemp fibres (Cannabis sativa), and three different biofabrication protocols were used during incubation, varying the time and surface treatment, including the addition of pre-colonised sawdust. The results indicate that density can be reduced by colonisation time, which will favourably impact thermal conductivity but will negatively affect vapour and liquid water control. Additionally, different fungi can exhibit different resistance to prolonged water absorption, and due to osmotic sensitivity, mycelium skin may also diminish moisture control. Finally, a collapse in the mycelium network after water immersion was observed through SEM, indicating how the microstructure is affected, which is also dependent on fungi species and the type of skin achieved. These results help to comprehend the differences and limitations of three of the most common species used for MBC fabrication and how precise engineering is needed to effectively control the material output.

Keywords: mycelium, thermal conductivity, vapor permeability, water absorption

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Authors: Jessica Achebe, Susan Tighe

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The reduction of material consumption and greenhouse gas emission when maintain and rehabilitating road networks can achieve added benefits including improved life cycle performance of pavements, reduced climate change impacts and human health effect due to less air pollution, improved productivity due to an optimal allocation of resources and reduced road user cost. This is the essence of incorporating environmental sustainability into pavement management. The functionality of performance measurement approach has made it one of the most valuable tool to Pavement Management Systems (PMSs) to account for different criteria in the decision-making process. However measuring the environmental performance of road network is still a far-fetched practice in road network management, more so an ostensive agency-wide environmental sustainability or sustainable maintenance specifications is missing. To address this challenge, this present research focuses on the environmental sustainability performance of network-level pavement management. The ultimate goal is to develop a framework to incorporate environmental sustainability in pavement management systems for network-level maintenance programming. In order to achieve this goal, this paper present the first step, the intention is to review the previous studies that employed environmental performance measures, as well as the suitability of environmental performance indicators for the evaluation of the sustainability of network-level pavement maintenance strategies. Through an industry practice survey, this paper provides a brief forward regarding the pavement manager motivations and barriers to making more sustainable decisions, and data needed to support the network-level environmental sustainability. The trends in network-level sustainable pavement management are also presented, existing gaps are highlighted, and ideas are proposed for network-level sustainable maintenance and rehabilitation programming.

Keywords: pavement management, environment sustainability, network-level evaluation, performance measures

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Authors: Clara Franch de la Cal, Christopher J Morris, Michael McArthur

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Cystic fibrosis (CF) is an autosomal recessive genetic disorder characterized by abnormal transport of chloride and sodium across the lung epithelium, leading to thick and viscous secretions. Within which CF patients suffer from repeated bacterial pulmonary infections, with Pseudomonas aeru-ginosa (PA) eliciting the greatest inflammatory response, causing an irreversible loss of lung func-tion that determines morbidity and mortality. The cell wall of PA is a permeability barrier to many antibacterials and the rise of Mutli-Drug Resistant strains (MDR) is eroding the efficacy of the few remaining clinical options. In addition when PA infection becomes established it forms an antibi-otic-resistant biofilm, embedded in which are slow growing cells that are refractive to drug treat-ment. Making the development of new antibacterials a major challenge. This work describes the development of new type of nanoparticulate oligonucleotide antibacterial capable of tackling PA infections, including MDR strains. It is being developed to both block growth and prevent biofilm formation. These oligonucleotide therapeutics, Transcription Factor Decoys (TFD), act on novel genomic targets by capturing key regulatory proteins to block essential bacterial genes and defeat infection. They have been successfully transfected into a wide range of pathogenic bacteria, both in vitro and in vivo, using a proprietary delivery technology. The surfactant used self-assembles with TFD to form a nanoparticle stable in biological fluids, which protects the TFD from degradation and preferentially transfects prokaryotic membranes. Key challenges are to adapt the nanoparticle so it is active against PA in the context of biofilms and to formulate it for administration by inhalation. This would allow the drug to be delivered to the respiratory tract, thereby achieving drug concentrations sufficient to eradicate the pathogenic organisms at the site of infection.

Keywords: antibacterials, transcriptional factor decoys (TFDs), pseudomonas aeruginosa

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Authors: Siti Anis Nadia Abu Bakar, Vani Tanggamani

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Over recent years, the quality and environmental management practices are the necessary tasks in hospitality industry in order to provide high quality services, a comfortable and safe environment for occupants as well as innovative nature and shareholders' satisfaction, its environmental and social added value sustainable. Numerous studies have observed and measured quality management program (QMProg) and environmental management practices (EMPrac) independently. This paper analyzed the level of QMProg, and EMPrac in hospitality industry, particularly on hotel performance, specifically in the context of Malaysia as hotel industry in Malaysia has contributed tremendously to the development in the Malaysia tourism industry.The research objectives are; (1) to analyze how the level of QMProg influences on firm performance; (2) to investigate the level of EMPrac and its influence on firm performance. This paper contributes to the literature by providing added-value to the service industry strategic decision-making processes by helping to predict the varying impacts of positive and negative corporate social responsibility (CSR) activities on financial performance in their respective industries. Further, this paper also contributes to develop more applicable CSR strategies. As a matter of fact, the findings of this paper has contributed towards an integrated management system that will assist a firm in implementation of their environmental strategy by creating a higher level of accountability for environmental performance. The best results in environmental systems have instigated managers to explore more options when dealing with problems, especially problems involving the reputation of their hotel. In conclusion, the results of the study infer that the best CSR strategies of the quality and environmental management practices influences hotel performance.

Keywords: corporate social responsibility (CSR), environmental management practices (EMPrac), performance (PERF), quality management program (QMProg)

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Authors: Taoufik Djennane

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Algeria is characterized by a fragmented language education policy. While pre-university education is entirely conducted in Arabic, higher education remains linguistically divided, with some fields offered in Arabic and others exclusively based on French. Within this linguistic policy, English remains far behind French. However, there has been a significant shift in the state’s linguistic orientation since the social riot of March 2019, known as El-Hirak, which ousted away the ex-president. Since then, social calls were voiced to get rid of French, and English started to receive an unprecedented political push. The historical decision only came at the beginning of the academic year 2023-2024 when the ministry of higher education imposed English as medium of instruction (hereafter EMI), especially in scientific and technological fields. As such, this paper considered this abrupt switch in the medium of instruction and its effects on the community of teachers. Building on a socio-psychological approach, teachers’ attitudes towards EMI were measured. Data were collected using classroom observation, semi-structured interviews and a survey. The results showed that a clear majority of teachers hold negative attitudes towards EMI. The point is that they are linguistically incompetent, and they are not ready yet to deliver content subjects in a language they have no, or little, command of. The study showed the importance of considering attitudes in the ‘policy-formation’ stage before the ‘implementation’ stage. The findings also proved that teachers are not passive bystanders; they can rather be the final arbiters imposing themselves as policy-makers resisting ministerial instructions through their linguistic practices inside the classroom which only acknowledge French. The study showed the necessity to avoid sudden switch and opt for gradual change, without putting aside those who are directly concerned with political/pedagogical measures (teachers, learners, etc).

Keywords: micro planning, EMI, language education policy, agency

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Authors: Ali Al Orf, Khawaja Bilal Waheed

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Background and objective: Joubert syndrome (JS) is a rare, autosomal-recessive condition. Early recognition is important for management and counseling. Magnetic resonance imaging (MRI) can help in diagnosis. Therefore, we sought to evaluate clinical presentation and MRI findings in Joubert syndrome and related disorders. Method: A retrospective review of genetically proven cases of Joubert syndromes and related disorders was reviewed for their clinical presentation, demographic information, and magnetic resonance imaging findings in a period of the last 10 years. Two radiologists documented magnetic resonance imaging (MRI) findings. The presence of hypoplasia of the cerebellar vermis with hypoplasia of the superior cerebellar peduncle resembling the “Molar Tooth Sign” in the mid-brain was documented. Genetic testing results were collected to label genes linked to the diagnoses. Results: Out of 12 genetically proven JS cases, most were females (9/12), and nearly all presented with hypotonia, ataxia, developmental delay, intellectual impairment, and speech disorders. 5/12 children presented at age of 1 or below. The molar tooth sign was seen in 10/12 cases. Two cases were associated with other brain findings. Most of the cases were found associated with consanguineous marriage Conclusion and discussion: The molar tooth sign is a frequent and reliable sign of JS and related disorders. Genes related to defective cilia result in malfunctioning in the retina, renal tubule, and neural cell migration, thus producing heterogeneous syndrome complexes known as “ciliopathies.” Other ciliopathies like Senior-Loken syndrome, Bardet Biedl syndrome, and isolated nephronophthisis must be considered as the differential diagnosis of JS. The main imaging findings are the partial or complete absence of the cerebellar vermis, hypoplastic cerebellar peduncles (giving MTS), and (bat-wing appearance) fourth ventricular deformity. LimitationsSingle-center, small sample size, and retrospective nature of the study were a few of the study limitations.

Keywords: Joubart syndrome, magnetic resonance imaging, molar tooth sign, hypotonia

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Authors: Daniel Asiamah Ameyaw

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This research attempts to investigate how lecturer evaluation makes a difference in enhancing higher education teaching and learning processes. The research questions to guide this research work states first as, “What are the perspectives on the difference made by evaluating academic teachers in order to enhance higher education teaching and learning processes?” and second, “What are the implications of the findings for Policy and Practice?” Data for this research was collected mainly through interviewing and partly documents review. Data analysis was conducted under the framework of grounded theory. The findings showed that for individual lecturer level, lecturer evaluation provides a continuous improvement of teaching strategies, and serves as source of data for research on teaching. At the individual student level, it enhances students learning process; serving as source of information for course selection by students; and by making students feel recognised in the educational process. At the institutional level, it noted that lecturer evaluation is useful in personnel and management decision making; it assures stakeholders of quality teaching and learning by setting up standards for lecturers; and it enables institutions to identify skill requirement and needs as a basis for organising workshops. Lecturer evaluation is useful at national level in terms of guaranteeing the competencies of graduates who then provide the needed manpower requirement of the nation. Besides, it mentioned that resource allocation to higher educational institution is based largely on quality of the programmes being run by the institution. The researcher concluded, that the findings have implications for policy and practice, therefore, higher education managers are expected to ensure that policy is implemented as planned by policy-makers so that the objectives can successfully be achieved.

Keywords: academic quality, higher education, lecturer evaluation, teaching and learning processes

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Authors: Adewale Tolulope Irewale, Elias Emeka Elemike, Christian O. Dimkpa, Emeka Emmanuel Oguzie

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As the global population steadily approaches the 10billion mark, the world is currently faced with two major challenges among others – accessing sustainable and clean energy, and food security. Accessing cleaner and sustainable energy sources to drive global economy and technological advancement, and feeding the teeming human population require sustainable, innovative, and smart solutions. To solve the food production problem, producers have relied on fertilizers as a way of improving crop productivity. Commercial inorganic fertilizers, which is employed to boost agricultural food production, however, pose significant ecological sustainability and economic problems including soil and water pollution, reduced input efficiency, development of highly resistant weeds, micronutrient deficiency, soil degradation, and increased soil toxicity. These ecological and sustainability concerns have raised uncertainties about the continued effectiveness of conventional fertilizers. With the application of nanotechnology, plant biomass upcycling offers several advantages in greener energy production and sustainable agriculture through reduction of environmental pollution, increasing soil microbial activity, recycling carbon thereby reducing GHG emission, and so forth. This innovative technology has the potential for a circular economy and creating a sustainable agricultural practice. Nanomaterials have the potential to greatly enhance the quality and nutrient composition of organic biomass which in turn, allows for the conversion of biomass into nanofertilizers that are potentially more efficient. Water hyacinth plant harvested from an inland water at Warri, Delta State Nigeria were air-dried and milled into powder form. The dry biomass were used to prepare biochar at a pre-determined temperature in an oxygen deficient atmosphere. Physicochemical analysis of the resulting biochar was carried out to determine its porosity and general morphology using the Scanning Transmission Electron Microscopy (STEM). The functional groups (-COOH, -OH, -NH2, -CN, -C=O) were assessed using the Fourier Transform InfraRed Spectroscopy (FTIR) while the heavy metals (Cr, Cu, Fe, Pb, Mg, Mn) were analyzed using Inductively Coupled Plasma – Optical Emission Spectrometry (ICP-OES). Impregnation of the biochar with nanonutrients were achieved under varied conditions of pH, temperature, nanonutrient concentrations and resident time to achieve optimum adsorption. Adsorption and desorption studies were carried out on the resulting nanofertilizer to determine kinetics for the potential nutrients’ bio-availability to plants when used as green fertilizers. Water hyacinth (Eichhornia crassipes) which is an aggressively invasive aquatic plant known for its rapid growth and profusion is being examined in this research to harness its biomass as a sustainable feedstock to formulate functionalized nano-biochar fertilizers, offering various benefits including water hyacinth biomass upcycling, improved nutrient delivery to crops and aquatic ecosystem remediation. Altogether, this work aims to create output values in the three dimensions of environmental, economic, and social benefits.

Keywords: biochar-based nanofertilizers, eichhornia crassipes, greener agriculture, sustainable ecosystem, water hyacinth

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Authors: Amin Naboureh, Ainong Li, Jinhu Bian, Guangbin Lei, Hamid Ebrahimy

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Large-scale land cover mapping has become a new challenge in land change and remote sensing field because of involving a big volume of data. Moreover, selecting the right classification method, especially when there are different types of landscapes in the study area is quite difficult. This paper is an attempt to compare the performance of different machine learning (ML) algorithms for generating a land cover map of the China-Central Asia–West Asia Corridor that is considered as one of the main parts of the Belt and Road Initiative project (BRI). The cloud-based Google Earth Engine (GEE) platform was used for generating a land cover map for the study area from Landsat-8 images (2017) by applying three frequently used ML algorithms including random forest (RF), support vector machine (SVM), and artificial neural network (ANN). The selected ML algorithms (RF, SVM, and ANN) were trained and tested using reference data obtained from MODIS yearly land cover product and very high-resolution satellite images. The finding of the study illustrated that among three frequently used ML algorithms, RF with 91% overall accuracy had the best result in producing a land cover map for the China-Central Asia–West Asia Corridor whereas ANN showed the worst result with 85% overall accuracy. The great performance of the GEE in applying different ML algorithms and handling huge volume of remotely sensed data in the present study showed that it could also help the researchers to generate reliable long-term land cover change maps. The finding of this research has great importance for decision-makers and BRI’s authorities in strategic land use planning.

Keywords: land cover, google earth engine, machine learning, remote sensing

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Authors: Jean Hugé, Francisco Benitez-Capistros, Giorgia Camperio-Ciani

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Addressing conservation conflicts requires the consideration of multiple stakeholders' perspectives and knowledge claims, in order to inform complex and possibly contentious decision-making dilemmas. Hence, a better understanding of why people in particular contexts act in a particular way in a conservation conflict is needed. First, this contribution aims at providing and applying an approach to map and interpret the diversity of subjective viewpoints with regard to iconic species in conservation conflicts. Secondly, this contribution aims to feed the reflection on the possible consequences of the diversity of perspectives for the future management of wildlife (in particular iconic species), based on case studies in Galapagos and Malaysia. The use of the semi-quantitative Q methodology allowed us to identify various perspectives on conservation in different social-ecological contexts. While the presence of iconic species may lead to a more passionate and emotional debate, it may also provide more opportunities for finding common ground and for jointly developing acceptable management solutions that will depolarize emergent, long-lasting or latent conservation conflicts. Based on the research team’s experience in the field, and on the integration of ecological and social knowledge, methodological and management recommendations are made with regard to conservation conflicts involving iconic wildlife. The mere presence of iconic wildlife does not guarantee its centrality in conservation conflicts, and comparisons will be drawn between the cases of the giant tortoises (Chelonoidis spec.) in Galapagos, Ecuador and the Milky Stork (Mycteria cinerea) in western peninsular Malaysia. Acknowledging the diversity of viewpoints, reflecting how different stakeholders see, act and talk about wildlife management, highlights the need to develop pro-active and resilient strategies to deal with these issues.

Keywords: conservation conflicts, Q methodology, Galapagos, Malaysia, giant tortoise, milky stork

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Authors: Hamidreza Joudaki, Yousefali Ziari

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The appearance of city and urbanism is one of the important processes which have affected social communities. Being industrialized urbanism developed along with each other in the history. In addition, they have had simple relationship for more than six thousand years, that is, from the appearance of the first cities. In 18th century by coming out of industrial capitalism, progressive development took place in urbanism in the world. In Iran, the city of each region made its decision by itself and the capital of region (downtown) was the only central part and also the regional city without any hierarchy, controlled its realm. However, this method of ruling during these three decays, because of changing in political, social and economic issues that have caused changes in rural and urban relationship. Moreover, it has changed the variety of performance of cities and systematic urban network in Iran. Today, urban system has very vast imbalanced apace and performance. In Isfahan, the trend of urbanism is like the other part of Iran and systematic urban hierarchy is not suitable and normal. This article is a quantitative and analytical. The statistical communities are Isfahan Province cities and the changes in urban network and its hierarchy during the period of fifty years (1956 -2006) has been surveyed. In addition, those data have been analyzed by model of Rank and size and Entropy index. In this article Iran cities and also the factor of entropy of primate city and urban hierarchy of Isfahan Province have been introduced. Urban residents of this Province have been reached from 55 percent to 83% (2006). As we see the analytical data reflects that there is mismatching and imbalance between cities. Because the entropy index was.91 in 1956.And it decreased to.63 in 2006. Isfahan city is the primate city in the whole of these periods. Moreover, the second and the third cities have population gap with regard to the other cities and finally, they do not follow the system of rank-size.

Keywords: urban network, urban hierarchy, primate city, Isfahan province, urbanism, first cities

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Authors: Romeu Bonk Mesquita

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Argentina is the South American number 1 country of destiny to intraregional migration flows. This research aims to shed light on the main trends of the Argentine immigrant policy from 2016 on, when Mauricio Marci was elected President, taking the approval of the current and fairly protective of human rights Ley de Migraciones (2003) as an analytical starting point. Foreign Policy Analysis (FPA) serves as the theoretical background, highlighting decision-making processes and institutional designs that encourage or constraint political and social actors. The analysis goes through domestic and international levels, observing how immigration policy is formulated as a public policy and is simultaneously connected to Mercosur and other international organizations, such as the International Organization for Migration (IOM) and the United Nations High Commissioner for Refugees (UNHCR). Thus, the study revolves around the Direccion Nacional de Migraciones, which is the state agency in charge of executing the country’s immigrant policy, as to comprehend how its internal processes and the connections it has with both domestic and international institutions shape Argentina’s immigrant policy formulation and execution. Also, it aims to locate the migration agenda within the country’s contemporary social and political context. The methodology is qualitative, case-based and oriented by process-tracing techniques. Empirical evidence gathered includes official documents and data, media coverage and interviews to key-informants. Recent events, such as the Decreto de Necesidad y Urgencia 70/2017 issued by President Macri, and the return of discursive association between migration and criminality, indicate a trend of nationalization and securitization of the immigration policy in contemporary Argentina.

Keywords: Argentine foreign policy, human rights, immigrant policy, Mercosur

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Authors: George B. Abaka-Wood, Massimiliano Zanin, Jonas Addai-Mensah, William Skinner

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The versatility of froth flotation has made it vital in the beneficiation of rare earth elements minerals from either high or low-grade ores. There has been a significant increase in the quantity of iron oxide silicate-rich tailings generated from the extraction of primary commodities such as copper and gold in Australia, which have been identified to contain very low-grade rare earth oxides (≤ 1%). There is a vast knowledge gap in the beneficiation of rare earth oxides from such tailings. The aim of this research is to investigate the feasibility of using fatty acids as collectors for the flotation recovery and upgrade of rare earth oxides from selected iron-oxide silicate-rich tailings. Two forms of fatty acid collectors (oleic acid and sodium oleate) were tested in this investigation. Flotation tests were carried out using a 1.2 L Denver D-12 cell. The effects of pulp pH, fatty acid dosage, particle size distribution (-150 +75 µm, -75 +38 µm and -38 µm) and conventional depressants (sodium silicate and starch) dosage on flotation recovery of rare earth oxides were investigated. A comparison of the flotation results indicated that sodium oleate was the more efficient fatty acid for rare earth oxides flotation at all the pulp pH investigated. The flotation performance was found to be particle size-dependent. Both sodium silicate and starch were unselective in decreasing the recovery of iron oxides and silicate minerals, respectively with the corresponding decrease in rare earth oxides recovery. Generally, iron oxides and silicate minerals formed the substantial fraction of the flotation concentrates obtained, both in the absence and presence of depressants, resulting in a generally low rare earth oxides upgrade, even though rare earth oxides recoveries were high. The flotation tests carried out on the tailings sample suggest the feasibility of rare earth oxides recovery using fatty acids, although particle size distribution and minerals liberation are key limiting factors in achieving selective rare earth oxides upgrade.

Keywords: depressants, flotation, oleic acid, sodium oleate

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Authors: Parimalakrishnan Sundararjan, Madheswaran Murugan, Dhanya Dharman, Yatindra Kumar, Sudhir Singh Gangwar, Guru Prasad Mohanta

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Antibiotic resistance AR is a global issue, India started to redress the issues of antibiotic resistance late and it plans to have: active surveillance of microbial resistance and promote appropriate use of antibiotics. The present study attempted to achieve appropriate use of antibiotics through pharmacists’ intervention at practice point. In a quasi-experimental prospective cohort study, the cases with bacteremia from four hospitals were identified during 2015 and 2016 for intervention. The pharmacists centered intervention: active screening of each prescription and comparing with the selection of antibiotics with susceptibility of the bacteria. Wherever irrationality noticed, it was brought to the notice of the treating physician for making changes. There were two groups: intervention group and control group without intervention. The active screening and intervention in 915 patients has reduced therapeutic regimen time in patients with bacteremia. The intervention group showed the decreased duration of hospital stay 3.4 days from 5.1 days. Further, multivariate modeling of patients who were in control group showed that patients in the intervention group had a significant decrease in both duration of hospital stay and infection-related mortality. Unlike developed countries, pharmacists are not active partners in patient care in India. This unique attempt of pharmacist’ invention was planned in consultation with hospital authorities which proved beneficial in terms of reducing the duration of treatment, hospital stay, and infection-related mortality. This establishes the need for a collaborative decision making among the health workforce in patient care at least for promoting rational use of antibiotics, an attempt to combat resistance.

Keywords: antibiotics resistance, intervention, bacteremia, multivariate modeling

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Authors: Diana C. El Assal, Fatima Monteiro, Caroline May, Peter Barbuti, Silvia Bolognin, Averina Nicolae, Hulda Haraldsdottir, Lemmer R. P. El Assal, Swagatika Sahoo, Longfei Mao, Jens Schwamborn, Rejko Kruger, Ines Thiele, Kathrin Marcus, Ronan M. T. Fleming

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Degeneration of substantia nigra pars compacta dopaminergic neurons is one of the hallmarks of Parkinson's disease. These neurons have a highly complex axonal arborisation and a high energy demand, so any reduction in ATP synthesis could lead to an imbalance between supply and demand, thereby impeding normal neuronal bioenergetic requirements. Synaptic mitochondria exhibit increased vulnerability to dysfunction in Parkinson's disease. After biogenesis in and transport from the cell body, synaptic mitochondria become highly dependent upon oxidative phosphorylation. We applied a systems biochemistry approach to identify the metabolic pathways used by neuronal mitochondria for energy generation. The mitochondrial component of an existing manual reconstruction of human metabolism was extended with manual curation of the biochemical literature and specialised using omics data from Parkinson's disease patients and controls, to generate reconstructions of synaptic and somal mitochondrial metabolism. These reconstructions were converted into stoichiometrically- and fluxconsistent constraint-based computational models. These models predict that Parkinson's disease is accompanied by an increase in the rate of glycolysis and a decrease in the rate of oxidative phosphorylation within synaptic mitochondria. This is consistent with independent experimental reports of a compensatory switching of bioenergetic pathways in the putamen of post-mortem Parkinson's disease patients. Ongoing work, in the context of the SysMedPD project is aimed at computational prediction of mitochondrial drug targets to slow the progression of neurodegeneration in the subset of Parkinson's disease patients with overt mitochondrial dysfunction.

Keywords: bioenergetics, mitochondria, Parkinson's disease, systems biochemistry

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Authors: Sanjay Kumar, Dinesh Pathak, Sudhir Saralch

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Copper oxide is a semiconductor that has been studied for several reasons such as the natural abundance of starting material copper (Cu); the easiness of production by Cu oxidation; their non-toxic nature and the reasonably good electrical and optical properties. Copper oxide is well-known as cuprite oxide. The cuprite is p-type semiconductors having band gap energy of 1.21 to 1.51 eV. As a p-type semiconductor, conduction arises from the presence of holes in the valence band (VB) due to doping/annealing. CuO is attractive as a selective solar absorber since it has high solar absorbency and a low thermal emittance. CuO is very promising candidate for solar cell applications as it is a suitable material for photovoltaic energy conversion. It has been demonstrated that the dip technique can be used to deposit CuO films in a simple manner using metallic chlorides (CuCl₂.2H₂O) as a starting material. Copper oxide films are prepared using a methanolic solution of cupric chloride (CuCl₂.2H₂O) at three baking temperatures. We made three samples, after heating which converts to black colour. XRD data confirm that the films are of CuO phases at a particular temperature. The optical band gap of the CuO films calculated from optical absorption measurements is 1.90 eV which is quite comparable to the reported value. Dip technique is a very simple and low-cost method, which requires no sophisticated specialized setup. Coating of the substrate with a large surface area can be easily obtained by this technique compared to that in physical evaporation techniques and spray pyrolysis. Another advantage of the dip technique is that it is very easy to coat both sides of the substrate instead of only one and to deposit otherwise inaccessible surfaces. This method is well suited for applying coating on the inner and outer surfaces of tubes of various diameters and shapes. The main advantage of the dip coating method lies in the fact that it is possible to deposit a variety of layers having good homogeneity and mechanical and chemical stability with a very simple setup. In this paper, the CuO thin films preparation by dip coating method and their characterization will be presented.

Keywords: absorber material, cupric oxide, dip coating, thin film

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Authors: Radmila Maksimovic, Sonja Ketin, Rade Biocanin, Jelena Maksimovic

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The abnormal conditions of life and work genetic factors often play a major role in incidence of diabetes-diabetes, heart disease and vascular disease, jaundice, and post traumatic stress. Trauma and post traumatic stress are most common in the displaced persons,and the focus of this paper is to shed light on this issue in former Yugoslavia, Yugoslavia and now in our country. This is caused by increased beta-cell sensitivity to viruses, the development of autoimmune antibodies against its own pancreascells, degenerative changes in cells that r esult in change of structure and insulin. In this paper, we dealt with traumatic events and long-term psycho social consequences for internally displaced persons, several years after displacement, and found a high level of PTSD symptoms. This stress is present in almost 1/3 of internally displaced persons, and every sixth person is suffering from PTSD in the past. Respondents generally suffer from symptoms of intrusion, but there was a large number of symptoms, avoidance and increased arousal. We also found that gender, age andeducation related to the symptoms. Females, and older respondents and internally displaced persons with lower levels of education how a higher level of PTSD symptoms, especially symptoms of intrusion and increase darousal. It is a highly traumatized sample in which more than 1/2 of respondents experienced more than three traumatic events in life,although the number of traumas experienced before, during and after the conflict varies.We found that during the war, internally displaced persons haveexperienced more traumatic events compared with the periodbefore and after the conflict. Trauma are different in type. No significant correlation between the number of experienced trauma and PTSD, suggesting that it is necessary to further study the structure of past traumas and the intermediary effects of certain risk factors and protective factors.

Keywords: living environment, displaced persons, jaundice, diabetes, trauma, diabetic hypertension, post-traumatic stress (PTSD), treatment

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Authors: Shaho Ghafur Ahmed

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The right to self-determination of peoples is a fundamental human right recognized by the principles of international law. It could be embodied in the internal level in the form of federalism. Most federal constitutions prevented the secession of constituent entities, while some remained silent, as the case of Iraq, and rare instances of them recognize it. But, after the failure of federalism, these entities seek to separate whenever the opportunity arises. In several cases, they have resort to peaceful methods in some others they resort to force. The constitutional Supreme Court, which guaranty the unity and integrity of the State, often prevent these attempts. After not a commitment of federalism in Iraq, which has been founded since 2004, the Kurdistan region, as the only federated entity, has conducted a unilateral referendum on 25 September 2017 for its independence. The Iraqi government refused it. The Iraqi Federal Supreme Court, through interpreting the constitutional provisions, decided that this referendum and it’s purposes, which was the independence of the region, was unconstitutional. Subsequently, the Iraqi government used forces and blockaded the region so as to force it to turn off this process. So, in this paper, the right to self-determination of the peoples in federated entities and its obstacles will be discussed through the comparative legal basis and analyzing the decisions of the Federal Constitutional Courts. We will compare the role that the Supreme Court of Canada played regarding the referendum that operated in Quebec in 1995, in which it refused only the unilaterally attempts for the independence of this province. While, in the case of the Kurdistan region, the Iraqi Federal Supreme Court has definitively refused this right. No measures were taken by this Court to protect the region from the Iraqi government reactions. This decision led to the questioning of the neutrality of this Court. So, from the point of view of the Kurdistan region, this Court became a political instrument to prevent it to be independent in the international community, in the absence of a clear constitutional provision, through an abstract and an incomplete interpretation of federal constitutional provisions.

Keywords: right of self-determination, federal supreme court, supremacy of federal constitution

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Authors: Mohammed Alassim

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In 2014, a UN survey stated that: "The greatest challenge to the adoption of whole-of government, which fundamentally rests on increased collaboration, is resistance to change among government actors". Change management has experienced both theoretically and practically many transformation over the years. When organizations have to implement radical changes, they have to encounter a plethora of issues which leads to ineffective or inefficient implementation of change in most cases. 70% of change projects fail because of human issues. It has been cited that” most studies still show a 60-70% failure rate for organizational change projects — a statistic that has stayed constant from the 1970’s to the present.”. E-government involves not just technical change but cultural, policy, social and organizational evolution. Managing change and overcoming resistance to change is seen as crucial in the success of E-government projects. Resistance can be from different levels in the organization (top management, middle management or employees at operational levels). There can be many reasons for resistance including fear of change and insecurity, lack of knowledge and absence of commitment from management to implement the change. The purpose of this study is to conduct in-depth research to understand the process of change and to identify the critical factors that have led to resistance from employees at different levels (top management, Middle management and operational employees) during e-government initiatives in the public sector in Saudi Arabia. The study is based on qualitative and empirical research methods conducted in the public sector in the Kingdom of Saudi Arabia. This research will use triangulation in data method (interview, group discussion and document review). This research will contribute significantly to knowledge in this field and will identify the measures that can be taken to reduce resistance to change, Upon analysis recommendations or model will be offered which can enable decision makers in public sector in Saudi Arabia how to plan, implement and evaluate change in e-government initiatives via change management strategy.

Keywords: change management, e-government, managing change, resistance to change

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Authors: Inayat Shah, Muhammad Omar Malik, Ghareeb Alshuwaier, Ronald H. Baxendale

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Background: The balance between angiogenesis and angiostasis is important in growth and developmental processes in the body. Angiogenic and angiostatic mediators control this balance. Endostatin is one of the prominent angiostatic mediators. The marked angiostatic effect of endostatin includes inhibiting endothelial cell migration, proliferation and apoptosis. Physical activity decreases the risk and development of many angiogenesis related health problems including atherosclerosis and numerous cancers. Physiological influences of different physical activities on plasma endostatin concentration are controversial and not completely clear. Moreover, correlation of physical characteristics and metabolic predictors during physical activity on circulating endostatin is indistinct and poorly speculated. The study aimed to determine the effects of mild, moderate and vigorous exercise on the concentration of endostatin in plasma. Methodology: 22 participants, 16 males (age = 30.6 ± 7.8 years) and 6 females (age = 26.5 ± 5 years) were recruited. Weekly session of different intensities exercise based on the predicted maximum heart of the participants [60%(low), 70% (moderate) and 80% (vigorous)] were carried out. The duration and work rate for each participant was determined through sub-maximal exercise. Standardization of the session was done on total energy expenditure of the participants per session. One pre exercise and two post exercise samples were taken at intervals of 10 and 60 minutes. Results: Pre-exercise mean endostatin was 101 ± 20 ng/dl. Low intensity exercise insignificantly decreased the endostatin concentration in plasma at 10 and 60 minutes 97 ± 20 ng/dl (p= 0.5), 98 ± 23 ng/dl (p= 0.8)). However, moderate (p= 0.022, 0.004) and vigorous intensities (p ≤ 0.001, 0.02) increased the endostatin concentrations significantly at both 10 and 60 minutes intervals respectively. The effects were not significantly influenced by gender, exercise mode (walking vs. running), components of exercise (HR, Speed, Gradients, distance, duration) or metabolism during exercise (VO₂ max, VCO₂, RER, energy expenditure, rate of carbohydrate or fats oxidation). Conclusion: Low intensity exercises did not influence endostatin concentration. However, moderate to high intensity exercises significantly increase endostatin concentration and may have potential benefits.

Keywords: angiogenesis, exercise, endostatin, physical activity

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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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Authors: Jayakant Singh, Enu Anand

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Background: Child marriage is widely prevalent in India. It is a form of gross human right violation that succumbs a child bride to be subservient to her husband within a marital relation. We investigated the relationship between age at marriage of women and her level of empowerment with physical violence experienced 12 months preceding the survey among young women aged 20-24 in two culturally distinct states- Bihar and Tamil Nadu of India. Methods: We used the information collected from 10514 young married women (20-24 years) at all India level, 373 in Bihar and 523 in Tamil Nadu from the third round of National Family Health Survey. Empowerment index was calculated using different parameters such as mobility, economic independence and decision making power of women using Principal Component Analysis method. Bivariate analysis was performed primarily using chi square for the test of significance. Logistic regression was carried out to assess the effect of age at marriage and empowerment on physical violence. Results: Lower level of women empowerment was significantly associated with physical violence in Tamil Nadu (OR=2.38, p<0.01) whereas child marriage (marriage before age 15) was associated with physical violence in Bihar (OR=3.27, p<0.001). The mean difference in age at marriage between those who experienced physical violence and those who did not experience varied by 7 months in Bihar and 10 months in Tamil Nadu. Conclusion: Culture specific intervention may be a key to reduction of violence against women as the results showed association of different factors contributing to physical violence in Bihar and Tamil Nadu. Marrying at an appropriate age perhaps is protective of abuse because it equips a woman to assert her rights effectively. It calls for an urgent consideration to curb both violence and child marriage with stricter involvement of family, civil society and the government. In the meanwhile physical violence may be recognized as a public health problem and integrate appropriate treatment to the victims within the health care institution.

Keywords: child marriage, empowerment, India, physical violence

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Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

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