Search results for: orphan nuclear receptors

Authors: S. Visetpotjanakit, C. Khrautongkieo

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Radiation monitoring in the environment and foodstuffs is one of the main responsibilities of Office of Atoms for Peace (OAP) as the nuclear regulatory body of Thailand. The main goal of the OAP is to assure the safety of the Thai people and environment from any radiological incidents. Various radioanalytical methods have been developed to monitor radiation and radionuclides in the environmental and foodstuff samples. To validate our analytical performance, several proficiency test exercises from the International Atomic Energy Agency (IAEA) have been performed. Here, the results of a proficiency test exercise referred to as the Proficiency Test for Tritium, Cobalt, Strontium and Caesium Isotopes in Seawater 2017 (IAEA-RML-2017-01) are presented. All radionuclides excepting ³H were analysed using various radioanalytical methods, i.e. direct gamma-ray counting for determining ⁶⁰Co, ¹³⁴Cs and ¹³⁷Cs and developed radiochemical techniques for analysing ¹³⁴Cs, ¹³⁷Cs using AMP pre-concentration technique and 90Sr using di-(2-ethylhexyl) phosphoric acid (HDEHP) liquid extraction technique. The analysis results were submitted to IAEA. All results passed IAEA criteria, i.e. accuracy, precision and trueness and obtained ‘Accepted’ statuses. These confirm the data quality from the OAP environmental radiation laboratory to monitor radiation in the environment.

Keywords: international atomic energy agency, proficiency test, radiation monitoring, seawater

Procedia PDF Downloads 171

Authors: Berengere Vire, Nicolas Salvetat, Yoann Lannay, Guillaume Marcellin, Siem Van Der Laan, Franck Molina, Dinah Weissmann

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Predicting suicidal behaviors is one of the most complex challenges of daily psychiatric practices. Today, suicide risk prediction using biological tools is not validated and is only based on subjective clinical reports of the at-risk individual. Therefore, there is a great need to identify biomarkers that would allow early identification of individuals at risk of suicide. Alterations of adenosine-to-inosine (A-to-I) RNA editing of neurotransmitter receptors and other proteins have been shown to be involved in etiology of different psychiatric disorders and linked to suicidal behavior. RNA editing is a co- or post-transcriptional process leading to a site-specific alteration in RNA sequences. It plays an important role in the epi transcriptomic regulation of RNA metabolism. On postmortem human brain tissue (prefrontal cortex) of depressed suicide victims, Alcediag found specific alterations of RNA editing activity on the mRNA coding for the serotonin 2C receptor (5-HT2cR). Additionally, an increase in expression levels of ADARs, the RNA editing enzymes, and modifications of RNA editing profiles of prime targets, such as phosphodiesterase 8A (PDE8A) mRNA, have also been observed. Interestingly, the PDE8A gene is located on chromosome 15q25.3, a genomic region that has recurrently been associated with the early-onset major depressive disorder (MDD). In the current study, we examined whether modifications in RNA editing profile of prime targets allow identifying disease-relevant blood biomarkers and evaluating suicide risk in patients. To address this question, we performed a clinical study to identify an RNA editing signature in blood of depressed patients with and without the history of suicide attempts. Patient’s samples were drawn in PAXgene tubes and analyzed on Alcediag’s proprietary RNA editing platform using next generation sequencing technology. In addition, gene expression analysis by quantitative PCR was performed. We generated a multivariate algorithm comprising various selected biomarkers to detect patients with a high risk to attempt suicide. We evaluated the diagnostic performance using the relative proportion of PDE8A mRNA editing at different sites and/or isoforms as well as the expression of PDE8A and the ADARs. The significance of these biomarkers for suicidality was evaluated using the area under the receiver-operating characteristic curve (AUC). The generated algorithm comprising the biomarkers was found to have strong diagnostic performances with high specificity and sensitivity. In conclusion, we developed tools to measure disease-specific biomarkers in blood samples of patients for identifying individuals at the greatest risk for future suicide attempts. This technology not only fosters patient management but is also suitable to predict the risk of drug-induced psychiatric side effects such as iatrogenic increase of suicidal ideas/behaviors.

Keywords: blood biomarker, next-generation-sequencing, RNA editing, suicide

Procedia PDF Downloads 259

Authors: K. Ravi, Sabu Subhash

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Densely compacted bentonite or bentonite–sand mixture has been identified as a suitable buffer in the deep geological repository (DGR) for the safe disposal of high-level nuclear waste (HLW) due to its favourable physicochemical and hydro-mechanical properties. The addition of sand to the bentonite enhances the thermal conductivity and compaction properties and reduces the drying shrinkage of the buffer material. The buffer material may undergo cyclic wetting and drying upon ingress of groundwater from the surrounding rock mass and from evaporation due to high temperature (50–210 °C) derived from the waste canister. The cycles of changes in temperature may result in thermal history, and the hydro-mechanical properties of the buffer material may be affected. This paper examines the influence of thermal history on the undrained shear strength of bentonite and bentonite-sand mixture. Bentonite from Rajasthan state and sand from the Assam state of India are used in this study. The undrained shear strength values are obtained by conducting unconfined compressive strength (UCS) tests on cylindrical specimens (dry densities 1.30 and 1.5 Mg/m3) of bentonite and bentonite-sand mixture consisting of 30 % bentonite+ 70 % sand. The specimens are preheated at temperatures varying from 50-150 °C for one, two and four hours in hot air oven. The results indicate that the undrained shear strength is increased by the thermal history of the buffer material. The specimens of bentonite-sand mixture exhibited more increase in strength compared to the pure bentonite specimens. This indicates that the sand content of the mixture plays a vital role in taking the thermal stresses of the bentonite buffer in DGR conditions.

Keywords: bentonite, deep geological repository, thermal history, undrained shear strength

Procedia PDF Downloads 345

Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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Authors: Hamsini Pulugurtha, V.S. Lakshmi Jagadmaba Paluri

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Distributed denial of service attack (DDoS) is one altogether the top-rated cyber threats presently. It runs down the victim server resources like a system of measurement and buffer size by obstructing the server to supply resources to legitimate shoppers. Throughout this text, we tend to tend to propose a mathematical model of DDoS attack; we discuss its relevancy to the choices like inter-arrival time or rate of arrival of the assault customers accessing the server. We tend to tend to further analyze the attack model in context to the exhausting system of measurement and buffer size of the victim server. The projected technique uses an associate in nursing unattended learning technique, self-organizing map, to make the clusters of identical choices. Lastly, the abstract applies mathematical correlation and so the standard likelihood distribution on the clusters and analyses their behaviors to look at a DDoS attack. These systems not exclusively interconnect very little devices exchanging personal data, but to boot essential infrastructures news standing of nuclear facilities. Although this interconnection brings many edges and blessings, it to boot creates new vulnerabilities and threats which might be conversant in mount attacks. In such sophisticated interconnected systems, the power to look at attacks as early as accomplishable is of paramount importance.

Keywords: application attack, bandwidth, buffer correlation, DDoS distribution flooding intrusion layer, normal prevention probability size

Procedia PDF Downloads 225

Authors: Belén Vinaixa Kinnear, Arturo Hidalgo López, Bernardo Elembo Wilasi, Pablo Fernández Pérez, Cecilia Hernández Fuentealba

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The expanded use of risk assessment in legislative and corporate decision-making has increased the role of expert judgement in giving data for security-related decision-making. Expert judgements are required in most steps of risk assessment: danger recognizable proof, hazard estimation, risk evaluation, and examination of choices. This paper presents a fault tree analysis (FTA), which implies a probabilistic failure analysis applied to leakage of oil in a subsea production system. In standard FTA, the failure probabilities of items of a framework are treated as exact values while evaluating the failure probability of the top event. There is continuously insufficiency of data for calculating the failure estimation of components within the drilling industry. Therefore, fuzzy hypothesis can be used as a solution to solve the issue. The aim of this paper is to examine the leaks from the Zafiro West subsea oil facility by using fuzzy fault tree analysis (FFTA). As a result, the research has given theoretical and practical contributions to maritime safety and environmental protection. It has been also an effective strategy used traditionally in identifying hazards in nuclear installations and power industries.

Keywords: expert judgment, probability assessment, fault tree analysis, risk analysis, oil pipelines, subsea production system, drilling, quantitative risk analysis, leakage failure, top event, off-shore industry

Procedia PDF Downloads 190

Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova

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The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.

Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF

Procedia PDF Downloads 265

Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

Procedia PDF Downloads 180

Authors: Derjew Ayele Ejigu, Houde Song, Xiaojing Liu

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This work presents the particle swarm optimization trained neural network (PSO-NN) supervisory proportional integral derivative (PID) control method to monitor the pressurized water reactor (PWR) core power for safe operation. The proposed control approach is implemented on the transfer function of the PWR core, which is computed from the state-space model. The PWR core state-space model is designed from the neutronics, thermal-hydraulics, and reactivity models using perturbation around the equilibrium value. The proposed control approach computes the control rod speed to maneuver the core power to track the reference in a closed-loop scheme. The particle swarm optimization (PSO) algorithm is used to train the neural network (NN) and to tune the PID simultaneously. The controller performance is examined using integral absolute error, integral time absolute error, integral square error, and integral time square error functions, and the stability of the system is analyzed by using the Bode diagram. The simulation results indicated that the controller shows satisfactory performance to control and track the load power effectively and smoothly as compared to the PSO-PID control technique. This study will give benefit to design a supervisory controller for nuclear engineering research fields for control application.

Keywords: machine learning, neural network, pressurized water reactor, supervisory controller

Procedia PDF Downloads 155

Authors: C. S. Wong, K. H. Badri, N. Ataollahi, K. P. Law, M. S. Su’ait, N. I. Hassan

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Novel bio-based polymer electrolyte was synthesized with LiClO4 as the main source of charge carrier. Initially, polyurethane-LiClO4 polymer electrolytes were synthesized via polymerization method with different NCO/OH ratios and labelled as PU1, PU2, PU3, and PU4. Subsequently, the chemical, thermal properties and ionic conductivity of the films produced were determined. Fourier transform infrared (FTIR) analysis indicates the co-ordination between Li+ ion and polyurethane in PU1 due to the greatest amount of hard segment of polyurethane in PU1 as proven by soxhlet analysis. The structures of polyurethanes were confirmed by 13 nuclear magnetic resonance spectroscopy (13C NMR) and FTIR spectroscopy. Differential scanning calorimetry (DSC) analysis indicates PU 1 has the highest glass transition temperature (Tg) corresponds to the most abundant urethane group which is the hard segment in PU1. Scanning electron microscopy (SEM) of the PU-LiClO4 shows the good miscibility between lithium salt and the polymer. The study found that PU1 possessed the greatest ionic conductivity (1.19 × 10-7 S.cm-1 at 298 K and 5.01 × 10-5 S.cm-1 at 373 K) and the lowest activation energy, Ea (0.32 eV) due to the greatest amount of hard segment formed in PU 1 induces the coordination between lithium ion and oxygen atom of carbonyl group in polyurethane. All the polyurethanes exhibited linear Arrhenius variations indicating ion transport via simple lithium ion hopping in polyurethane. This research proves the NCO content in polyurethane plays an important role in affecting the ionic conductivity of this polymer electrolyte.

Keywords: ionic conductivity, palm kernel oil-based monoester-OH, polyurethane, solid polymer electrolyte

Procedia PDF Downloads 425

Authors: Hossein Sabouri

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News stories as one of the branches of political discourse has always been regarded a sensitive and challenging area. Political translators often encounter some struggles that are vitally important when it comes to deal with the political tension between the source culture and the target one. Translating news stories is of prime importance since it has widespread availability and power of defining or even changing the facts. News translation is usually more than straight transfer of source text. Like original text endeavoring to manipulate the readers’ minds with imposing their ideologies, translated text seeking to change these ideologies influenced by ideological power. In other words, translation product is not considered more than a recontextualisation of the source text. The present study examines possible criteria for occurring changes in the translation process of news stories based on the ideological and political stance of translator using theories of ‘critical discourse analysis’and ‘translation and power. Fairclough investigates the ideological issues in (political) discourse and Tymoczko studies the political and power-related engagement of the translator in the process of translation. Incorporation of Fairclough and Gentzler and Tymoczko’s theories paves the way for the researcher to looks at the ideological power position of the translator. Data collection and analysis have been accomplished using 17 political-text samples taken from online news agencies which are related to the ‘Iran’s Nuclear Program’. Based on the findings, recontextualisation is mainly observed in terms of the strategies of ‘substitution, omissions, and addition’ in the translation process. The results of the study suggest that there is a significant relationship between the translation of political texts and ideologies of target culture.

Keywords: news translation, recontextualisation, ideological power, political discourse

Procedia PDF Downloads 193

Authors: Lola T. Alimkhodjaeva, Lola T. Zakirova, Soniya S. Ziyavidenova

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Background: Breast cancer (BC) is still one of the urgent oncology problems. The essential obstacle to the full anti-tumor therapy implementation is drug resistance development. Taking into account the fact that chemotherapy is main antitumor treatment in BC patients, the important task is to improve treatment results. Certain success in overcoming this situation has been associated with the use of methods of extracorporeal blood treatment (ECBT), plasmapheresis. Materials and Methods: We examined 129 women with resistant BC stages 3-4, aged between 56 to 62 years who had previously received 2 courses of CAF chemotherapy. All patients additionally underwent 2 courses of CAF chemotherapy but against the background ECBT with ultrasonic exposure. We studied the following parameters: 1. The highlights of peripheral blood before and after therapy. 2. The state of cellular immunity and identification of activation markers CD23 +, CD25 +, CD38 +, CD95 + on lymphocytes was performed using monoclonal antibodies. Evaluation of humoral immunity was determined by the level of main classes of immunoglobulins IgG, IgA, IgM in serum. 3. The degree of tumor regression was assessed by WHO recommended 4 gradations. (complete - 100%, partial - more than 50% of initial size, process stabilization–regression is less than 50% of initial size and tumor advance progressing). 4. Medical pathomorphism in the tumor was determined by Lavnikova. 5. The study of immediate and remote results, up to 3 years and more. Results and Discussion: After performing extracorporeal blood treatment anemia occurred in 38.9%, leukopenia in 36.8%, thrombocytopenia in 34.6%, hypolymphemia in 26.8%. Studies of immunoglobulin fractions in blood serum were able to establish a certain relationship between the classes of immunoglobulin A, G, M and their functions. The results showed that after treatment the values of main immunoglobulins in patients’ serum approximated to normal. Analysis of expression of activation markers CD25 + cells bearing receptors for IL-2 (IL-2Rα chain) and CD95 + lymphocytes that were mediated physiological apoptosis showed the tendency to increase, which apparently was due to activation of cellular immunity cytokines allocated by ultrasonic treatment. To carry out ECBT on the background of ultrasonic treatment improved the parameters of the immune system, which were expressed in stimulation of cellular immunity and correcting imbalances in humoral immunity. The key indicator of conducted treatment efficiency is the immediate result measured by the degree of tumor regression. After ECBT performance the complete regression was 10.3%, partial response - 55.5%, process stabilization - 34.5%, tumor advance progressing no observed. Morphological investigations of tumor determined therapeutic pathomorphism grade 2 in 15%, in 25% - grade 3 and therapeutic pathomorphism grade 4 in 60% of patients. One of the main criteria for the effect of conducted treatment is to study the remission terms in the postoperative period (up to 3 years or more). The remission terms up to 3 years with ECBT was 34.5%, 5-year survival was 54%. Carried out research suggests that a comprehensive study of immunological and clinical course of breast cancer allows the differentiated approach to the choice of methods for effective treatment.

Keywords: breast cancer, immunoglobulins, extracorporeal blood treatment, chemotherapy

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Authors: Kassa Endalamaw Rewnu

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The variability properties of a quasar sample, spectroscopically complete to magnitude J = 22.0, are investigated on a time baseline of 2 years using three different photometric bands (U, J and F). The original sample was obtained using a combination of different selection criteria: colors, slitless spectroscopy and variability, based on a time baseline of 1 yr. The main goals of this work are two-fold: first, to derive the percentage of variable quasars on a relatively short time baseline; secondly, to search for new quasar candidates missed by the other selection criteria; and, thus, to estimate the completeness of the spectroscopic sample. In order to achieve these goals, we have extracted all the candidate variable objects from a sample of about 1800 stellar or quasi-stellar objects with limiting magnitude J = 22.50 over an area of about 0.50 deg2. We find that > 65% of all the objects selected as possible variables are either confirmed quasars or quasar candidates on the basis of their colors. This percentage increases even further if we exclude from our lists of variable candidates a number of objects equal to that expected on the basis of `contamination' induced by our photometric errors. The percentage of variable quasars in the spectroscopic sample is also high, reaching about 50%. On the basis of these results, we can estimate that the incompleteness of the original spectroscopic sample is < 12%. We conclude that variability analysis of data with small photometric errors can be successfully used as an efficient and independent (or at least auxiliary) selection method in quasar surveys, even when the time baseline is relatively short. Finally, when corrected for the different intrinsic time lags corresponding to a fixed observed time baseline, our data do not show a statistically significant correlation between variability and either absolute luminosity or redshift.

Keywords: nuclear activity, galaxies, active quasars, variability

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Authors: Seokyoon Moon, Yun-Ho Ahn, Heejoong Kim, Sujin Hong, Yunseok Lee, Youngjune Park

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Gas hydrate is a non-stoichiometric crystalline compound consisting of host water-framework and low molecular weight guest molecules. Small gaseous molecules such as CH₄, CO₂, and N₂ can be captured in the host water framework lattices of the gas hydrate with specific temperature and pressure conditions. The three well-known crystal structures of structure I (sI), structure II (sII), and structure H (sH) are determined by the size and shape of guest molecules. In this study, we measured the phase equilibria of binary (2,2-dimethyl-1-propanol + CO₂, CH₄, N₂) hydrates to explore their fundamental thermodynamic characteristics. We identified the structure of the binary gas hydrate by employing synchrotron high-resolution powder diffraction (HRPD), and the guest distributions in the lattice of gas hydrate were investigated via dispersive Raman and ¹³C solid-state nuclear magnetic resonance (NMR) spectroscopies. The end-to-end distance of 2,2-dimethyl-1-propanol was calculated to be 7.76 Å, which seems difficult to be enclathrated in large cages of sI or sII. However, due to the flexibility of the host water framework, binary hydrates of sI or sII types can be formed with the help of small gas molecule. Also, the synchrotron HRPD patterns revealed that the binary hydrate structure highly depends on the type of help gases; a cubic Fd3m sII hydrate was formed with CH₄ or N₂, and a cubic Pm3n sI hydrate was formed with CO₂. Interestingly, dispersive Raman and ¹³C NMR spectra showed that the unique tuning phenomenon occurred in binary (2,2-dimethyl-1-propanol + CO₂) hydrate. By optimizing the composition of NPA, we can achieve both thermodynamic stability and high CO₂ storage capacity for the practical application to CO₂ capture.

Keywords: clathrate, gas hydrate, neopentyl alcohol, CO₂, tuning phenomenon

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Authors: A. Kumar, V. Devadas

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Energy is the basic element for any country’s economic development. India is one of the most populated countries, and is dependent on fossil fuel and nuclear-based energy generation. The energy sector faces huge challenges and is dependent on the import of energy from neighboring countries to fulfill the gap in demand and supply. India has huge setbacks for efficient energy generation, distribution, and consumption, therefore they consume more quantity of energy to produce the same amount of Gross Domestic Product (GDP) compared to the developed countries. Technology and technique use, availability, and affordability in the various sectors are varying according to their economic status. In this paper, an attempt is made to quantify the domestic electrical energy consumption in Jodhpur, India. Survey research methods have been employed and stratified sampling technique-based households were chosen for conducting the investigation. Pre-tested survey schedules are used to investigate the grassroots level study. The collected data are analyzed by employing statistical techniques. Thereafter, a multiple regression model is developed to understand the functions of total electricity consumption in the domestic sector corresponding to other independent variables including electrical appliances, age of the building, household size, education, etc. The study resulted in identifying the governing variable in energy consumption at the household level and their relationship with the efficiency of household-based electrical and energy appliances. The analysis is concluded with the recommendation for optimizing the gap in peak electrical demand and supply in the domestic sector.

Keywords: appliance, consumption, electricity, households

Procedia PDF Downloads 116

Authors: Taufik Hidayat, Yudha Nurhantari, Bambang U. D. Rianto

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Introduction: The examination of the Short Tandem Repeats (STR) locus on nuclear DNA is very useful in solving the paternity cases. The purpose of this study is to know the description of paternity cases and paternity index/probability of paternity analysis based on Indonesian allele frequency at Sardjito Hospital Yogyakarta. Method: This was an observational study with cross-sectional analytic method. Population and sample were all cases of disputed paternity from January 2011 to June 2015 that fulfill the inclusion and exclusion criteria and were examined at Forensic Medicine Unit of Sardjito Hospital, Medical Faculty of Gadjah Mada University. The paternity index was calculated with EasyDNA Program by Fung (2013). Analysis of the study was conducted by comparing the results through unpaired categorical test using Kolmogorov-Smirnov test. This study was designed with 95% confidence interval (CI) with α = 5% and significance level is p < 0,05. Results: From 42 disputed paternity cases we obtained trio paternity cases were 32 cases (76.2%) and duo without a mother was 10 cases (23.8%). The majority of the fathers' estimated ages were 21-30 years (33.3%) and the mother's age was 31-40 years (38.1%). The majority of the ages of children examined for paternity were under 12 months (47.6%). The majority of ethnic clients are Javanese. Conclusion of inclusion was 57.1%, and exclusion was 42.9%. The Kolmogorov-Smirnov test obtained p-value = 0.673. Conclusion: There is no significant difference between paternity index/probability of paternity based on Indonesian allele frequency between trio and duo of paternity.

Keywords: disputed paternity, paternity index, probability of paternity, short tandem

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Authors: Yakui Bai, Chen Sun, Ke Wang

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An experimental investigation of environmental effect on fatigue behavior in SA508 Gr.3 Cl.2 Steam Generator Forging CAP1400 nuclear power plant has been carried out. In order to simulate actual loading condition, a range of strain amplitude was applied in different low cycle fatigue (LCF) tests. The current American Society of Mechanical Engineers (ASME) design fatigue code does not take full account of the interactions of environmental, loading, and material's factors. A range of strain amplitude was applied in different low cycle fatigue (LCF) tests at a strain rate of 0.01%s⁻¹. A design fatigue model was constructed by taking environmentally assisted fatigue effects into account, and the corresponding design curves were given for the convenience of engineering applications. The corrosion fatigue experiment was performed in a strain control mode in 320℃ borated and lithiated water environment to evaluate the effects of a mixed environment on fatigue life. Stress corrosion cracking (SCC) in steam generator large forging in primary water of pressurized water reactor was also observed. In addition, it is found that the CF life of SA508 Gr.3 Cl.2 decreases with increasing temperature in the water environment. The relationship between the reciprocal of temperature and the logarithm of fatigue life was found to be linear. Through experiments and subsequent analysis, the mechanisms of reduced low cycle fatigue life have been investigated for steam generator forging.

Keywords: failure behavior, low alloy steel, steam generator forging, stress corrosion cracking

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Authors: Parinaz Tavakoli Zaniani, Dimitrios Balomenos

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Mitochondrial reactive oxygen species (mROS) play a crucial role in macrophage pro-inflammatory activation, although a detailed understanding of the mechanism and kinetics by which mROS drive signaling molecules is still lacking. In general, it is thought that NF-κB activation drives mROS and general ROS production. Here, We performed a detailed kinetic analysis of mROS production during macrophage activation. We found early mROS generation after LPS (lipopolysaccharide) stimulation. Remarkably as early as 5 minutes, mROS signaling promoted initial NF-κB, MAPK activation and pro-inflammatory cytokine production, as established through inhibition or quenching of mROS. On the contrary, NF-κB inhibition had no effect on mROS production. Our findings point to a mechanism by which mROS increase TRAF-6 ubiquitination and, thus NF-κB activity. mROS inhibition reduced LPS-induced lethality in an in vivo septic shock model by controlling pro-inflammatory cytokine production. Overall, our research provides novel insights into the role of mROS as a primary messenger in the pathway of macrophage and as a regulator of inflammatory responses. We found that early mROS production promotes initial NF-κB, and MAPK activation by regulating TRAF-6 ubiquitination and that mROS inhibition can reduce LPS-induced inflammatory cytokines and lethality in a septic shock model. These findings might lead to novel immunotherapeutic strategies targeting early mROS production and control of extreme inflammation in the context of sepsis and other inflammatory diseases.

Keywords: mitochondria, reactive oxygen species, nuclear factor κB, lipopolysaccharide, macrophages

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Authors: Massimo Zucchetti

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In the US, the SPARC-ARC projects of compact tokamaks are being developed: both are aimed at the technological demonstration of fusion power reactors with cutting-edge technology but following different design approaches. However, they show more similarities than differences in the fuel cycle, safety, radiation protection, environmental, waste and decommissioning aspects: all reactors, either experimental or demonstration ones, have to fulfill certain "essential" requirements to pass from virtual to real machines, to be built in the real world. The paper will discuss these "essential" requirements. Some of the relevant activities in these fields, carried out by our research group (ESSENTIAL group), will be briefly reported, with the aim of showing some methodology aspects that have been developed and might be of wider interest. Also, a non-competitive comparison between our results for different projects will be included when useful. The question of advanced D-He3 fuel cycles to be used for those machines will be addressed briefly. In the past, the IGNITOR project of a compact high-magnetic field D-T ignition experiment was found to be able to sustain limited D-He3 plasmas, while the Candor project was a more decisive step toward D-He3 fusion reactors. The following topics will be treated: Waste management and radioactive safety studies for advanced fusion power plants; development of compact high-field advanced fusion reactors; behavior of nuclear materials under irradiation: neutron-induced radioactivity due to side DT reactions, radiation damage; accident analysis; reactor siting.

Keywords: advanced fuel fusion reactors, deuterium-helium3, high-field tokamaks, fusion safety

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Authors: Johannes Bibinger, Sebastian Eibl, Hans-Joachim Gudladt

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This study considers the influence of different irradiation scenarios on the thermal degradation of carbon fiber-reinforced polymers (CFRP). Real threats are simulated, such as fires with long-lasting low heat fluxes and nuclear heat flashes with short-lasting high heat fluxes. For this purpose, coated and uncoated quasi-isotropic samples of the commercially available CFRP HexPly® 8552/IM7 are thermally irradiated from one side by a cone calorimeter and a xenon short-arc lamp with heat fluxes between 5 and 175 W/cm² at varying time intervals. The specimen temperature is recorded on the front and backside as well as at different laminate depths. The CFRP is non-destructively tested with ultrasonic testing, infrared spectroscopy (ATR-FTIR), scanning electron microscopy (SEM), and micro-focused computed X-Ray tomography (μCT). Destructive tests are performed to evaluate the mechanical properties in terms of interlaminar shear strength (ILSS), compressive and tensile strength. The irradiation scenarios vary significantly in heat flux and exposure time. Thus, different heating rates, radiation effects, and temperature distributions occur. This leads to unequal decomposition processes, which affect the sensitivity of the strength type and damage behaviour of the specimens. However, with the use of surface coatings, thermal degradation of composite materials can be delayed.

Keywords: CFRP, one-sided thermal damage, high heat flux, heating rate, non-destructive and destructive testing

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Authors: Yong Jae Yu, Jae-Yeol Cho

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In the design of structural concrete for impact loading, design or model codes often employ a dynamic increase factor (DIF) to impose dynamic effect on static response. Dynamic increase factors that are obtained from laboratory material test results and that are commonly given as a function of strain rate only are quite different from each other depending on the design concept of design codes like ACI 349M-06, fib Model Code 2010 and ACI 370R-14. Because the dynamic increase factors currently adopted in the codes are too simple and limited to consider a variety of strength of materials, their application in practical design is questionable. In this study, the dynamic increase factors used in the three codes were validated through the finite element analysis of reinforced concrete slab elements which were tested and reported by other researcher. The test was intended to simulate a wall element of the containment building in nuclear power plants that is assumed to be subject to impact scenario that the Pentagon experienced on September 11, 2001. The finite element analysis was performed using the ABAQAUS 6.10 and the plasticity models were employed for the concrete, reinforcement. The dynamic increase factors given in the three codes were applied to the stress-strain curves of the materials. To estimate the dynamic increase factors, strain rate was adopted as a parameter. Comparison of the test and analysis was done with regard to perforation depth, maximum deflection, and surface crack area of the slab. Consequently, it was found that DIF has so great an effect on the behavior of the reinforced concrete structures that selection of DIF should be very careful. The result implies that DIF should be provided in design codes in more delicate format considering various influence factors.

Keywords: impact, strain rate, DIF, slab elements

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Authors: Jefri Draup, Antoine Ambard, Chi-Toan Nguyen

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Zirconium alloys exhibit solid-state phase transformations under thermal loading. These can lead to a significant evolution of the microstructure and associated mechanical properties of materials used in nuclear fuel cladding structures. Therefore, the ability to capture effects of phase transformation on the material constitutive behavior is of interest during conditions of severe transient thermal loading. Whilst typical Avrami, or Johnson-Mehl-Avrami-Kolmogorov (JMAK), type models for phase transformations have been shown to have a good correlation with the behavior of Zircaloy-4 under constant heating rates, the effects of variable and fast heating rates are not fully explored. The present study utilises the results of in-situ high energy synchrotron X-ray diffraction (SXRD) measurements in order to validate the phase transformation models for Zircaloy-4 under fast variable heating rates. These models are used to assess the performance of fuel cladding structures under loss of coolant accident (LOCA) scenarios. The results indicate that simple Avrami type models can provide a reasonable indication of the phase distribution in experimental test specimens under variable fast thermal loading. However, the accuracy of these models deteriorates under the faster heating regimes, i.e., 100Cs⁻¹. The studies highlight areas for improvement of simple Avrami type models, such as the inclusion of temperature rate dependence of the JMAK n-exponent.

Keywords: accident, fuel, modelling, zirconium

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Authors: Vinod Kumar

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Background: Tissue Plasminogen Activator (tPA) showed a level 1 benefit in acute stroke (within 3-6 hrs). Intracarotid sodium nitroprusside (ICSNP) has been studied in this context with a wide treatment window, fast recovery and affordability. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a)Retrograde Neurotransmission (acute cases): 1)Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2)Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b)Vasospasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c)Long-Term Potentıatıon (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Des’gn: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat stroke cases. Case-control prospective study. Mater’als And Methods: The experimental population included 82 stroke patients (10 patients were given control treatments without superfusion or with 5% dextrose superfusion, and 72 patients comprised the ICSNP group). The mean time for superfusion was 9.5 days post-stroke. Pre- and post-ICSNP status was monitored by NIHSS, MRI and TCD. Results: After 90 seconds in the ICSNP group, the mean change in the NIHSS score was a decrease of 1.44 points, or 6.55%; after 2 h, there was a decrease of 1.16 points; after 24 h, there was an increase of 0.66 points, 2.25%, compared to the control-group increase of 0.7 points, or 3.53%; at 7 days, there was an 8.61-point decrease, 44.58%, compared to the control-group increase of 2.55 points, or 22.37%; at 2 months in ICSNP, there was a 6.94-points decrease, 62.80%, compared to the control-group decrease of 2.77 points, or 8.78%. TCD was documented and improvements were noted. Conclusions: ICSNP is a swift-acting drug in the treatment of stroke, acting within 90 seconds on day 9.5 post-stroke with a small decrease after 24 hours. The drug recovers from this decrease quickly.

Keywords: brain infarcts, intracarotid sodium nitroprusside, perforators, vasodilatıons, retrograde transmission, the 10, 000-fold effect

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Authors: Zoe Goldberger, Priscilla S. Briquez, Jeffrey A. Hubbell

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Tumor cells have mutations resulting from genetic instability that the immune system can actively recognize. Immune checkpoint immunotherapy (ICI) is commonly used in the clinic to re-activate immune reactions against mutated proteins, called neoantigens, resulting in tumor remission in cancer patients. However, only around 20% of patients show durable response to ICI. While tumor mutational burden (TMB) has been approved by the Food and Drug Administration (FDA) as a criterion for ICI therapy, the relevance of the subcellular localizations of the mutated proteins within the tumor cell has not been investigated. Here, we hypothesized that localization of mutations impacts the effect of immune responsiveness to ICI. We analyzed publicly available tumor mutation sequencing data of ICI treated patients from 3 independent datasets. We extracted the subcellular localization from the UniProtKB/Swiss-Prot database and quantified the proportion of membrane, cytoplasmic, nuclear, or secreted mutations per patient. We analyzed this information in relation to response to ICI treatment and overall survival of patients showing with 1722 ICI-treated patients that high mutational burden localized at the membrane (mTMB), correlate with ICI responsiveness, and improved overall survival in multiple cancer types. We anticipate that our results will ameliorate predictability of cancer patient response to ICI with potential implications in clinical guidelines to tailor ICI treatment. This would not only increase patient survival for those receiving ICI, but also patients’ quality of life by reducing the number of patients enduring non-effective ICI treatments.

Keywords: cancer, immunotherapy, membrane neoantigens, efficacy prediction, biomarkers

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Authors: Arjun, Gautam, Sanjeev Naval

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Concrete technology has been changing rapidly and constantly since its discovery. Concrete is the most widely used man-made construction material, versatility of making concrete is the 2nd largest consumed material on earth. In this paper an effort has been made to use metal aggregates in concrete has been discussed, the metal aggregates has been named as “BAUT” which had outstandingly qualities to resist shear, tension and compression forces. In this paper, COARSE BAUT AGGREGATES (C.B.A.) 10mm & 20mm and FINE BAUT AGGREGATES (F.B.A.) 3mm were divided and used for making high performance concrete (H.P.C). This “BAUT” had cutting edge technology through draft and design by the use of Auto CAD, ANSYS software can be used effectively In this research paper we study high performance concrete (H.P.C) with “BAUT” and consider the grade of M65 and finally we achieved the result of 90-95 Mpa (high compressive strength) for mega structures and irregular structures where center of gravity (CG) is not balanced. High Performance BAUT Concrete is the extraordinary qualities like long-term performance, no sorptivity by BAUT AGGREGATES, better rheological, mechanical and durability proportion that conventional concrete. This high strength BAUT concrete using “BAUT” is applied in the construction of mega structure like skyscrapers, dam, marine/offshore structures, nuclear power plants, bridges, blats and impact resistance structures. High Performance BAUT Concrete which is a controlled concrete possesses invariable high strength, reasonable workability and negligibly permeability as compare to conventional concrete by the mix of Super Plasticizers (SMF), silica fume and fly ash.

Keywords: BAUT, High Strength Concrete, High Performance Concrete, Fine BAUT Aggregate, Coarse BAUT Aggregate, metal aggregates, cutting edge technology

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Authors: Nuri Başpınar, Abdullah Başoğlu, Özgür Özdemir, Çağlayan Özel, FundaTerzi, Özgür Yaman

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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like nonalcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family were performed. LDL-cholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARα, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to precede in this effectiveness.

Keywords: high protein and energy diet, boron, metabolomics, transcriptomic

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Authors: Shahid Iqbal

Abstract:

It is observed that international migration is increased over a time period in all over the world counties and Pakistan is not escaped. It played a pivotal role in household economy and affecting the gender roles both positive and negative in the developing countries. A vast majority of males from Pakistan migrate to other countries in search of employment and income generating activities and their families left behind, particularly nuclear families are subjected to different social problems. In this scenario, most of the responsibilities lie on the female partners as they have to play the role of male as well as female for their children and household chores. Wives of some males feel loneliness, isolation and a sense of insecurity. Keeping in mind, these realities, this study aims to explore the social impact of husbands’ absentee ship on the lives of families left behind. Particularly, wives’ own experiences will be analyzed. This study will be carried out in the District Gujrat of Punjab Pakistan. Since this study will focus on observing the social impact of male’s migration on families, so, all the households that had at least one member abroad will be the potential respondents of the study. Purposes sampling technique will be used as method to locate the respondents. A focus group discussion will be conducted as a tool for the collection of data. Those women will be approached who are taking care of their families in the absence of their husbands for last three years. For the sake of analysis Interpretative Phenomenological Analysis (IPA) will be applied and researcher will explore how participants make sense of their personal experience and social world.

Keywords: social consequences, male migration, left behind, absentee ship, Pakistan

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Authors: G. Hosseini, A. Sadighzadeh, M. Rahimnejad, N. Hosseini, Z. Jamalzadeh

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Metal pollutants, especially heavy metals from anthropogenic sources such as metallurgical industries’ waste including mining, smelting, casting or production of nuclear fuel, including mining, concentrate production and uranium processing ends in the environment contamination (water and soil) and risk to human health around the facilities of this type of industrial activity. There are different methods that can be used to remove these contaminants from water and soil. These are very expensive and time-consuming. In this case, the people have been forced to leave the area and the decontamination is not done. For example, in the case of Chernobyl accident, an area of 30 km around the plant was emptied of human life. A very efficient and cost-effective method for decontamination of the soil and the water is phytoremediation. In this method, the plants preferentially native plants which are more adaptive to the regional climate are well used. In this study, three types of plants including Alfalfa, Sunflower and wheat were used to Barium decontamination. Alfalfa and Sunflower were not grown good enough in Saghand mine’s soil sample. This can be due to non-native origin of these plants. But, Wheat rise in Saghand Uranium Mine soil sample was satisfactory. In this study, we have investigated the effect of 4 types of acids inclusive nitric acid, oxalic acid, acetic acid and citric acid on the removal efficiency of Barium by Wheat. Our results indicate the increase of Barium absorption in the presence of citric acid in the soil. In this paper, we will present our research and laboratory results.

Keywords: phytoremediation, heavy metal, wheat, soil

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Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz

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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.

Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes

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Authors: Farrah Putri Salmanida, Mei-Li Wu, Rika Wahyuningtyas, Wen-Bin Chung, Hso-Chi Chaung, Ko-Tung Chang

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Within the field of immunotherapy, oncolytic virotherapy (OVT) employs dual approaches that directly eliminate tumor cells while preserving healthy ones and indirectly reprogram the tumor microenvironment (TME) to elicit antitumor responses. Within the TME, tumor associated macrophages (TAMs) manifest characteristics akin to those of anti-inflammatory M2 macrophages, thus earning the designation of M2-like TAMs. In prior research, two antigens denoted as A1 (g6Ld10T) and A3 (ORF6L5), derived from a complete sequence of ORF5 with partial sequence of ORF6 in Porcine Reproductive and Respiratory Syndrome Virus Genotype 2 (PRRSV-2), demonstrated the capacity to repolarize M2-type porcine alveolar macrophages (PAMs) into M1 phenotypes. In this study, we sought for utilizing OVT strategies by introducing A1 or A3 on TAMs to endow them with the anti-tumor traits of M1 macrophages while retaining their capacity to target cancer cells. Upon exposing human THP-1-derived M2 macrophages to a cross-species test with 2 µg/ml of either A1 or A3 for 24 hours, real time PCR revealed that A3, but not A1, treated cells exhibited upregulated gene expressions of M1 markers (CCR7, IL-1ß, CCL2, Cox2, CD80). These cells reacted to virus-derived antigen, as evidenced by increased expression of pattern-recognition receptors TLR3, TLR7, and TLR9, subsequently providing feedback in the form of type I interferon responses like IFNAR1, IFN-ß, IRF3, IRF7, OAS1, Mx1, and ISG15. Through an MTT assay, only after 15 µg/ml of A3 treatment could the cell viability decrease, with a predicted IC50 of 16.96 µg/ml. Interestingly, A3 caused dose-dependent toxicity to a rat C6 glial cancer cell line even at doses as low as 2.5 µg/ml and reached its IC50 at 9.419 µg/ml. Using Annexin V/7AAD staining and PCR test, we deduced that a significant proportion of C6 cells were undergoing the early apoptosis phase predominantly through the intrinsic apoptosis cascade involving Bcl-2 family proteins. Following this stage, we conducted a test on A3’s repolarization ability, which revealed a significant rise in M1 gene expression markers, such as TNF, CD80, and IL-1ß, in M2-like TAMs generated in vitro from murine RAW264.7 macrophages grown with conditioned medium of 4T1 breast cancer cells. This was corroborated by the results of transcriptome analysis, which revealed that the primary subset among the top 10 to top 30 significantly upregulated differentially expressed genes (DEGs) dominantly consisted of M1 macrophages profiles, including Ccl3, Ccl4, Csf3, TNF, Bcl6b, Stc1, and Dusp2. Our findings unveiled the remarkable potential of the PRRSV-derived antigen A3 to repolarize macrophages while also being capable of selectively inducing apoptosis in cancerous cells. While further in vivo study is needed for A3, it holds promise as an adjuvant by its dual effects in cancer therapy modalities.

Keywords: cancer cell apoptosis, interferon responses, macrophage repolarization, recombinant protein

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