Search results for: tandem and ovoid

Authors: Khaled Khleifat, Muayyad Abboud, Amjad Khleifat, Humodi Saeed

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In this study, Pseudomonas aeruginosa was isolated from infected burn patients and characterized by standard biochemical tests. The in vitro copper uptake was compared between this isolated pathogenic strain and two non-pathogenic control strains of Gram positive bacteria Bacillusthuringiensis strain Israelisas well as Gram negative bacteria Enterobacter aerogenes. Maximum copper uptake of 470 ppm/g biomass was obtained by P. aeruginosa strain, while the control strains B. thuringiensis andEnterobacter aerogenes had copper uptake of 350 and 383 ppm/g biomass, respectively. However, the lowest copper uptake (60 ppm/g biomass) was observed with another control the saprophytic strain Pseudomonas (Shewanella) putrefaciens. A further investigation regarding the effect of copper toxicity on bacterial growth, gave an MIC score of 600 ppm for P. aeruginosa strain compared to 460 and 300 ppm for the two Gram positive and Gram negative control strains, respectively. In tandem with these in vitro findings, blood analysis on burn patients infected with P. aeruginosa has indicated a selective decrease of copper (hypocupremia) and ceruloplasmin plasma levels. The iron metabolism was also affected by this copper deprivation leading to a similar decrease in plasma levels of PCV, iron, total iron binding capacity, and transferrin. All these hematological changes were significantly different (P < 0.05) from the matched group of non-infected burn patients. The observed hypocupremia in infected burn patients was attributed to demanding scavenger ability by P. aeruginosa strain for the copper of plasma.

Keywords: pseudomonas, Cu uptake, burn patients, biosorption

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Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.

Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon

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Authors: Rodrigo Pacheco-Ruiz, Jonathan Adams, Felix Pedrotti

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This paper introduces the applicability of underwater photogrammetric survey within challenging conditions as the main tool to enhance and enrich the process of documenting archaeological excavation through the creation of 4D models. Photogrammetry was being attempted on underwater archaeological sites at least as early as the 1970s’ and today the production of traditional 3D models is becoming a common practice within the discipline. Photogrammetry underwater is more often implemented to record exposed underwater archaeological remains and less so as a dynamic interpretative tool.  Therefore, it tends to be applied in bright environments and when underwater visibility is > 1m, reducing its implementation on most submerged archaeological sites in more turbid conditions. Recent years have seen significant development of better digital photographic sensors and the improvement of optical technology, ideal for darker environments. Such developments, in tandem with powerful processing computing systems, have allowed underwater photogrammetry to be used by this research as a standard recording and interpretative tool. Using multi-source photogrammetry (5, GoPro5 Hero Black cameras) this paper presents the accumulation of daily (4D) underwater surveys carried out in the Early Bronze Age (3,300 BC) to Late Ottoman (17th Century AD) archaeological site of Ropotamo in the Bulgarian Black Sea under challenging conditions (< 0.5m visibility). It proves that underwater photogrammetry can and should be used as one of the main recording methods even in low light and poor underwater conditions as a way to better understand the complexity of the underwater archaeological record.

Keywords: 4D modelling, Black Sea Maritime Archaeology Project, multi-source photogrammetry, low visibility underwater survey

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Authors: Adriano Mortada

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Human history is rife with conflict, and the question of refugee status determination and their rehabilitation has been up for debate since. Refugee Status Determination is the administrative or legal process by which UNHCR or governments determine whether a person seeking international protection or asylum can be identified as a refugee under international, regional, or national law. Refugee Status Determination is considered to be a vital process in aiding refugees’ realization of their rights under international law. One of the major reasons why the refugee status determination is considered an “issue”, and is one that is much debated upon annually, is the fact that the national bureaucratic systems are rigid and unbending. This is particularly concerning in the 21st century despite human advancement in policy and diplomacy, working in tandem with the United Nations and their charters and resolutions on human rights and dignity. The paper seeks to criticize the European member states' response to the refugee crisis and their inflexible and prejudiced bureaucratic systems when it comes to refugee status determination. The paper looks at multiple case studies as primary evidence and the alternate case studies where the system helped refugees, like those in Jordan, Pakistan, Turkey, and Lebanon. The main concern of the paper is to highlight the bias in the selected European systems, which do not stem from the Human Rights Charter but rather on the basis of geographical backgrounds, cultural and religious affiliations of those seeking refugee status or asylum in their respective countries. The paper hopes to not only create awareness about this issue but also provide a research background to advocacy programs to bring a change in the systems.

Keywords: refugee status determination, human rights, bureaucracy, United Nations, European Union

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Authors: Shivaji M. Sonawane, N. B. Chaure

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ZnTe is direct band gap, the P-type semiconductor with the high absorption coefficient of the order of 104cm-1 is suitable for solar cell development. It can be used as a low resistive ohmic contact to CdS/CdTe or tandem solar cell application. ZnTe and Cu-ZnTe thin film have been electrochemically synthesized on to fluorine-doped tin oxide coated glass substrates using three electrode systems containing Ag/AgCl, graphite and FTO as reference, counter and working electrode respectively were used to deposit the thin films. The aqueous electrolytic solution consist of 0.5M TeO2, 0.2M ZnSO4, and 0.1M Na3C6H5O7:2H2O, 0.1MC6H8O7:H2O and 0.1mMCuSO4 with PH 2.5 at room temperature was used. The reaction mechanism is studied in the cyclic voltammetry to identify the deposition potentials of ZnTe and Cu-ZnTe.The potential was optimized in the range -0,9 to -1,1 V. Vs Ag/AgCl reference electrode. The effect of deposition potential on the structural properties was studied by using X-ray diffraction. The X-ray diffraction result reveled cubic crystal structure of ZnTe with preferential (111) orientation with cubic structure. The surface morphology and film composition were analyzed by means of Scanning electron microscopy (SEM) and Energy Dispersive Analysis of X- Rays (EDAX). The optical absorption measurement has been analyzed for the band gap determination of deposited layers about 2.26 eV by UV-Visible spectroscopy. The drastic change in resistivity has been observed due to incorporation of copper probably due to the diffusion of Cu into grain boundaries.

Keywords: ohmic back contact, zinc telluride, electrodeposition, photovoltaic devices

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Authors: Nataša Stojić, Mira Pucarević, Nebojša Ralević, Vojislava Bursić, Gordan Stojić

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Surface soil (0 – 10 cm) samples from 52 sampling sites along the length of railway tracks on the territory of Srem (the western part of the Autonomous Province of Vojvodina, itself part of Serbia) were collected and analyzed for 7 polychlorinated biphenyls (PCBs) in order to see how the distance from the railroad on the one hand and dump on the other hand, affect the concentration of PCBs (CPCBs) in the soil. Samples were taken at a distance of 0.03 to 4.19 km from the railway and 0.43 to 3.35 km from the landfills. For the soil extraction the Soxhlet extraction (USEPA 3540S) was used. The extracts were purified on a silica-gel column (USEPA 3630C). The analysis of the extracts was performed by gas chromatography with tandem mass spectrometry. PCBs were not detected only at two locations. Mean total concentration of PCBs for all other sampling locations was 0,0043 ppm dry weight (dw) with a range of 0,0005 to 0,0227 ppm dw. On the part of the data that were interesting for this research with statistical methods (PCA) were isolated factors that affect the concentration of PCBs. Data were also analyzed using the Pearson's chi-squared test which showed that the hypothesis of independence of CPCBs and distance from the railway can be rejected. Hypothesis of independence between CPCB and the percentage of humus in the soil can also be rejected, in contrast to dependence of CPCB and the distance from the landfill where the hypothesis of independence cannot be rejected. Based on these results can be said that railway transport is a potential source of PCBs. The next step in this research is to establish the position of transformers which are located near sampling sites as another important factor that affects the concentration of PCBs in the soil.

Keywords: GC/MS, landfill, PCB, railway, soil

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Authors: László Forró

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The organolead halide perovskite CH3NH3PbI3 and its derivatives are known to be very efficient light harvesters revolutionizing the field of solid-state solar cells. The major research area in this field is photovoltaic device engineering although other applications are being explored, as well. Recently, we have shown that nanowires of this photovoltaic perovskite can be synthesized which in association with carbon nanostructures (carbon nanotubes and graphene) make outstanding composites with rapid and strong photo-response. They can serve as conducting electrodes, or as central components of detectors. The performance of several miniature devices based on these composite structures will be demonstrated. Our latest findings on the guided growth of perovskite nanowires by solvatomorph graphoepitaxy will be presented. This method turned out to be a fairly simple approach to overcome the spatially random surface nucleation. The process allows the synthesis of extremely long (centimeters) and thin (a few nanometers) nanowires with a morphology defined by the shape of nanostructured open fluidic channels. This low-temperature solution-growth method could open up an entirely new spectrum of architectural designs of organometallic-halide-perovskite-based heterojunctions and tandem solar cells, LEDs and other optoelectronic devices. Acknowledgment: This work is done in collaboration with Endre Horvath, Massimo Spina, Alla Arakcheeva, Balint Nafradi, Eric Bonvin1, Andrzej Sienkievicz, Zsolt Szekrenyes, Hajnalka Tohati, Katalin Kamaras, Eduard Tutis, Laszlo Mihaly and Karoly Holczer The research is supported by the ERC Advanced Grant (PICOPROP670918).

Keywords: photovoltaics, perovskite, nanowire, photodetector

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Authors: Ali Mahmoud, Shadi Najjar, Mounir Mabsout, Kassim Tarhini

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Empirical expressions for estimating the wheel load distribution and live-load bending moment are typically specified in highway bridge codes such as the AASHTO procedures. The purpose of this paper is to analyze the reliability levels that are inherent in reinforced concrete slab bridges that are designed based on the simplified empirical live load equations in the AASHTO LRFD procedures. To achieve this objective, bridges with multi-lanes (three and four lanes) and different spans are modeled using finite-element analysis (FEA) subjected to HS20 truck loading, tandem loading, and standard lane loading per AASHTO LRFD procedures. The FEA results are compared with the AASHTO LRFD moments in order to quantify the biases that might result from the simplifying assumptions adopted in AASHTO. A reliability analysis is conducted to quantify the reliability index for bridges designed using AASHTO procedures. To reach a consistent level of safety for three- and four-lane bridges, following a previous study restricted to one- and two-lane bridges, the live load factor in the design equation proposed by AASHTO LRFD will be assessed and revised if needed by alternating the live load factor for these lanes. The results will provide structural engineers with more consistent provisions to design concrete slab bridges or evaluate the load-carrying capacity of existing bridges.

Keywords: reliability analysis of concrete bridges, finite element modeling, reliability analysis, reinforced concrete bridge design, load carrying capacity

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Authors: Monika Szymańska-Czerwińska, Agnieszka Jodełko, Krzysztof Niemczuk

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Q fever (coxiellosis) is an infectious disease of animals and humans causes by C. burnetii and widely distributed throughout the world. Cattle and small ruminants are commonly known as shedders of C. burnetii. The aims of this study were the evaluation of seroprevalence and shedding of C. burnetii in cattle. Genotypes of the pathogen present in the tested specimens were also identified using MLVA (Multiple Locus Variable-Number Tandem Repeat Analysis) and MST (multispacer sequence typing) methods. Sampling was conducted in different regions of Poland in 2018-2021. In total, 2180 bovine serum samples from 801 cattle herds were tested by ELISA (enzyme-linked immunosorbent assay). 489 specimens from 157 cattle herds such as: individual milk samples (n=407), bulk tank milk (n=58), vaginal swabs (n=20), placenta (n=3) and feces (n=1) were subjected to C. burnetii specific qPCR. The qPCR (IS1111 transposon-like repetitive region) was performed using Adiavet COX RealTime PCR kit. Genotypic characterization of the strains was conducted utilizing MLVA and MST methods. MLVA was performed using 6 variable loci. The overall herd-level seroprevalence of C. burnetii infection was 36.74% (801/2180). Shedders were detected in 29.3% (46/157) cattle herds in all tested regions. ST 61 sequence type was identified in 10 out of 18 genotyped strains. Interestingly one strain represents sequence type which has never been recorded previously. MLVA method identified three previously known genotypes: most common was J but also I and BE were recognized. Moreover, a one genotype has never been described previously. Seroprevalence and shedding of C. burnetii in cattle is common and strains are genetically diverse.

Keywords: Coxiella burnetii, cattle, MST, MLVA, Q fever

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Authors: Manal Suleiman, George Abu-Aqil, Uraib Sharaha, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman, Mahmoud Huleihel

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Klebsiella pneumoniae is one of the most aggressive multidrug-resistant bacteria associated with human infections resulting in high mortality and morbidity. Thus, for an effective treatment, it is important to diagnose both the species of infecting bacteria and their susceptibility to antibiotics. Current used methods for diagnosing the bacterial susceptibility to antibiotics are time-consuming (about 24h following the first culture). Thus, there is a clear need for rapid methods to determine the bacterial susceptibility to antibiotics. Infrared spectroscopy is a well-known method that is known as sensitive and simple which is able to detect minor biomolecular changes in biological samples associated with developing abnormalities. The main goal of this study is to evaluate the potential of infrared spectroscopy in tandem with Random Forest and XGBoost machine learning algorithms to diagnose the susceptibility of Klebsiella pneumoniae to antibiotics within approximately 20 minutes following the first culture. In this study, 1190 Klebsiella pneumoniae isolates were obtained from different patients with urinary tract infections. The isolates were measured by the infrared spectrometer, and the spectra were analyzed by machine learning algorithms Random Forest and XGBoost to determine their susceptibility regarding nine specific antibiotics. Our results confirm that it was possible to classify the isolates into sensitive and resistant to specific antibiotics with a success rate range of 80%-85% for the different tested antibiotics. These results prove the promising potential of infrared spectroscopy as a powerful diagnostic method for determining the Klebsiella pneumoniae susceptibility to antibiotics.

Keywords: urinary tract infection (UTI), Klebsiella pneumoniae, bacterial susceptibility, infrared spectroscopy, machine learning

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Authors: Nabil Jallouli, Luisa M. Pastrana-Martínez, Ana R. Ribeiro, Nuno F. F. Moreira, Joaquim L. Faria, Olfa Hentati, Adrián M. T. Silva, Mohamed Ksibi

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Degradation and mineralization of ibuprofen (IBU) were investigated using Ultraviolet (UV) Light Emitting Diodes (LEDs) in TiO2 photocatalysis. Samples of ultrapure water (UP) and a secondary treated effluent of a municipal wastewater treatment plant (WWTP), both spiked with IBU, as well as a highly concentrated IBU (230 mgL-1) pharmaceutical industry wastewater (PIWW), were tested in the TiO2/UV-LED system. Three operating parameters, namely, pH, catalyst load and number of LEDs were optimized. The process efficiency was evaluated in terms of IBU removal using high performance liquid chromatography (HPLC) and ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Additionally, the mineralization was investigated by determining the dissolved organic carbon (DOC) content. The chemical structures of transformation products were proposed based on the data obtained using liquid chromatography with a high resolution mass spectrometer ion trap/time-of-flight (LC-MS-IT-TOF). A possible pathway of IBU degradation was accordingly proposed. Bioassays were performed using the marine bacterium Vibrio fischeri to evaluate the potential acute toxicity of original and treated wastewaters. TiO2 heterogeneous photocatalysis was efficient to remove IBU from UP and from PIWW, and less efficient in treating the wastewater from the municipal WWTP. The acute toxicity decreased by ca. 40% after treatment, regardless of the studied matrix.

Keywords: acute toxicity, Ibuprofen, UV-LEDs, wastewaters

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Authors: Guoqian Ren, Haijiang Li, Jisong Zhang

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Over the past 40 years, urban development has undergone large-scale, high-speed expansion, beyond what was previously considered normal and in a manner not proportionally related to population growth or physical considerations. With more scientific and refined decision-making in the urban construction process, new urbanization approaches, aligned with public-private partnerships (PPPs) which evolved in the early 1990s, have become acceptable and, in some situations, even better solutions to outstanding urban municipal construction projects, especially in developing countries. However, as the main driving force to deal with urban public services, PPPs are still problematic regarding value for money (VFM) process in most large-scale construction projects. This paper therefore reviews recent PPP articles in popular project management journals and relevant toolkits, published in the last 10 years, to identify the indicators that influence VFM within PPPs across regions. With increasing concerns about profitability and environmental and social impacts, the current PPP structure requires a more integrated platform to manage multi-performance project life cycles. Building information modelling (BIM), a popular approach to the procurement process in AEC sectors, provides the potential to ensure VFM while also working in tandem with the semantic approach to holistically measure life cycle costs (LCC) and achieve better sustainability. This paper suggests that BIM applied to the entire PPP life cycle could support holistic decision-making regarding VFM processes and thus meet service targets.

Keywords: public-private partnership, value for money, building information modelling, semantic approach

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Authors: Leila Najafian

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Sarcoplasmic protein hydrolysates (SPHs) and myofibrillar protein hydrolysates (MPHs) from patin (Pangasius sutchi) were produced using two types of proteases: Papain and Alcalase. 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) radical scavenging activities and metal chelating activity assays for antioxidant activities were carried out on the SPHs and MPHs. The hydrolysates were isolated and purified by ultrafiltration, gel filtration and reverse phase high-performance liquid chromatography (RP-HPLC) and liquid chromatography with tandem mass spectrometry detection (LC-MS/MS) was used in identifying peptide sequences. The results showed that when the DH of MPHs increased, the protein solubility increased, while the highest amount of the protein solubility of SPHs was after 60 min incubation. The effect of DH on antioxidant activities of SPHs and MPHs was investigated. Among the hydrolysates, papain-MPH and Alcalase-SPH, which had the highest antioxidant activities, were purified. The potent fractions obtained from RP-HPLC of sarcoplasmic (SI 3 fraction) and myofibrillar (MI 4 fraction) hydrolysates showed the highest DPPH radical scavenging activity. The FVNQPYLLYSVHMK peptide for MPH and the LVVDIPAALQHA peptide for SPH exhibited the highest antioxidant activity. The presence of hydrophobic and hydrophilic amino acids, namely leucine (L), valine (V), phenylalanine (F), histidine (H) and proline (P), in the peptide sequences of SPH and MPH are believed to contribute to high antioxidant activity. Hence, SPH and MPH from patin have the potential as a natural functional ingredient in food and pharmaceutical industry.

Keywords: patin (Pangasius sutchi), protein hydrolysates, antioxidative peptides, mass spectrometry

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Authors: Ana Vulic, Tina Lesic, Nina Kudumija, Maja Kis, Manuela Zadravec, Nada Vahcic, Tomaz Polak, Jelka Pleadin

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Mycotoxins are toxic secondary metabolites produced by numerous types of molds. They can contaminate both food and feed so that they represent a serious public health concern. Production of dry-fermented meat products involves ripening, during which molds can overgrow the product surface, produce mycotoxins, and consequently contaminate the final product. Citrinin is a mycotoxin produced mainly by the Penicillium citrinum. Data on citrinin occurrence in both food and feed are limited. Therefore, there is a need for research on citrinin occurrence in these types of meat products. The LC-MS/MS method for citrinin determination was developed and validated. Sample preparation was performed using immunoaffinity columns, which resulted in clean sample extracts. Method validation included the determination of the limit of detection (LOD), the limit of quantification (LOQ), recovery, linearity, and matrix effect in accordance to the latest validation guidance. The determined LOD and LOQ were 0.60 µg/kg and 1.98 µg/kg, respectively, showing a good method sensitivity. The method was tested for its linearity in the calibration range of 1 µg/L to 10 µg/L. The recovery was 100.9 %, while the matrix effect was 0.7 %. This method was employed in the analysis of 47 samples of dry-fermented sausages collected from local households. Citrinin wasn’t detected in any of these samples, probably because of the short ripening period of the tested sausages that takes three months tops. The developed method shall be used to test other types of traditional dry-cured products, such as prosciuttos, whose surface is usually more heavily overgrown by surface molds due to the longer ripening period.

Keywords: citrinin, dry-fermented meat products, LC-MS/MS, mycotoxins

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Authors: Lily Yarney, Emmanuel M. Y. Seidu, Thomas Chireh Kuusaanu, Belinda Adzimah-Yeboah

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Healthcare delivery is labor-intensive; the role of the health worker is, therefore, indispensable in maintaining and improving individual and population health. In Ghana, doctor-patient ratio is 1:10,450, with a disproportionate tilt in favor of the relatively resource rich southern part of the country. The Upper West Region located in Northern Ghana, is among the poorest regions in the country. The study was aimed at finding out the reasons why medical doctors are unwilling to accept postings to the Upper West Region where their services are needed most despite some efforts to attract, motivate and retain them. Current initiatives by the Ministry of Health and its partners to attract and retain doctors in the region were also examined. Qualitative methodology was employed with an in-depth interview guide to collect data. Sixteen respondents comprising medical doctors, health managers, and other health-related partners purposively selected took part in the study. Data were recorded, transcribed, coded, and categorized into themes in tandem with the objectives of the study. The study found that medical doctors are unwilling to take up appointments in the Upper West Region because of limited opportunities for career and continuing professional development, poor financial inducement, and weak leadership, among other important contextual social and cultural factors. Critical success factors to surmount these challenges include concessions and sponsorship for medical specialization training for doctors and clear implementable national and local policies on postings.

Keywords: attraction, retention, medical doctors, deprived regions, Ghana

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Authors: Javier Romera, Alberto Justo, Ignacio Fidalgo, Joshue Perez, Javier Araluce

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Nowadays, individuals with mobility needs face a significant challenge when docking vehicles. In many cases, after parking, they encounter insufficient space to exit, leading to two undesired outcomes: either avoiding parking in that spot or settling for improperly placed vehicles. To address this issue, the following paper presents a parking control system employing gestural teleoperation. The system comprises three main phases: capturing body markers, interpreting gestures, and transmitting orders to the vehicle. The initial phase is centered around the MediaPipe framework, a versatile tool optimized for real-time gesture recognition. MediaPipe excels at detecting and tracing body markers, with a special emphasis on hand gestures. Hands detection is done by generating 21 reference points for each hand. Subsequently, after data capture, the project employs the MultiPerceptron Layer (MPL) for indepth gesture classification. This tandem of MediaPipe's extraction prowess and MPL's analytical capability ensures that human gestures are translated into actionable commands with high precision. Furthermore, the system has been trained and validated within a built-in dataset. To prove the domain adaptation, a framework based on the Robot Operating System (ROS), as a communication backbone, alongside CARLA Simulator, is used. Following successful simulations, the system is transitioned to a real-world platform, marking a significant milestone in the project. This real vehicle implementation verifies the practicality and efficiency of the system beyond theoretical constructs.

Keywords: gesture detection, mediapipe, multiperceptron layer, robot operating system

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Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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Authors: Viveka Jackson, Erouscilla Joseph, Denise Beckles, Thomas Christopher

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Sulphur Dioxide (SO2) has been defined as a non-flammable, non-explosive, colourless gas, having a pungent, irritating odour, and is one of the main gases emitted from volcanoes. Sulphur dioxide has been recorded in concentrations hazardous to humans (0.25 – 0.5 ppm (~650 – 1300 μg/m3), downwind of many volcanoes and hence warrants constant air-quality monitoring around these sites. It has been linked to an increase in chronic respiratory disease attributed to long-term exposures and alteration in lung and other physiological functions attributed to short-term exposures. Sulphur Springs in Saint Lucia is a highly active geothermal area, located within the Soufrière Volcanic Centre, and is a park widely visited by tourists and locals. It is also a current source of continuous volcanic emissions via its many fumaroles and bubbling pools, warranting concern by residents and visitors to the park regarding the effects of exposure to these gases. In this study, we introduce a novel SO2 measurement system for the monitoring and quantification of ambient levels of airborne volcanic SO2 using low-cost technology. This work involves the extensive production of low-cost SO2 monitors/samplers, as well as field examination in tandem with standard commercial samplers (SO2 diffusion tubes). It also incorporates community involvement in the volcanic monitoring process as non-professional users of the instrument. We intend to present the preliminary monitoring results obtained from the low-cost samplers, to identify the areas in the Park exposed to high concentrations of ambient SO2, and to assess the feasibility of the instrument for non-professional use and application in volcanic settings

Keywords: ambient SO2, community-based monitoring, risk-reduction, sulphur springs, low-cost

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Authors: Jin Young Kim, Kwon Kyoo Kang

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The SGR1 (STAYGREEN1) protein is a critical regulator of plant leaves in chlorophyll degradation and senescence. The functions and mechanisms of tomato SGR1 action are poorly understood and worthy of further investigation. To investigate the function of the SGR1 gene, we generated a SGR1-knockout (KO) null line via clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and conducted RNA sequencing and gas chromatography tandem mass spectrometry (GC-MS/MS) analysis to identify the differentially expressed genes. The SlSGR1 (Solanum lycopersicum SGR1) knockout null line clearly showed a turbid brown color with significantly higher chlorophyll and carotenoid content compared to wild-type (WT) fruit. Differential gene expression analysis revealed 728 differentially expressed genes (DEGs) between WT and sgr1 #1-6 line, including 263 and 465 downregulated and upregulated genes, respectively, for which fold change was >2, and the adjusted p-value was <0.05. Most of the DEGs were related to photosynthesis and chloroplast function. In addition, the pigment, carotenoid changes in sgr1 #1-6 line was accumulated of key primary metabolites such as sucrose and its derivatives (fructose, galactinol, raffinose), glycolytic intermediates (glucose, G6P, Fru6P) and tricarboxylic acid cycle (TCA) intermediates (malate and fumarate). Taken together, the transcriptome and metabolite profiles of SGR1-KO lines presented here provide evidence for the mechanisms underlying the effects of SGR1 and molecular pathways involved in chlorophyll degradation and carotenoid biosynthesis.

Keywords: tomato, CRISPR/Cas9, null line, RNA-sequencing, metabolite profiling

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Authors: Spyridon Komaitis, Christos Koutsarnakis, Evangelos Drosos, Aristotelis Kalyvas

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This study opts to investigate the intrinsic architecture, morphology, and spatial relationship of the subcortical pathways implicated in the connectivity of the motor/premotor cortex and SMA/pre-SMA complex. Twenty normal, adult, formalin-fixed cerebral hemispheres were explored through the fiber micro-dissection technique. Lateral to medial and medial to lateral dissections focused on the area of interest were performed in a tandem manner and under the surgical microscope. We traced the subcortical architecture, spatial relationships, and axonal connectivity of four major pathways: a) the dorsal component of the SLF (SLF-I) was found to reside in the medial aspect of the hemisphere and seen to connect the precuneus with the SMA and pre-SMA complex, b) the frontal longitudinal system (FLS) was consistently encountered as the natural anterior continuation of the SLF-II and SLF-III and connected the premotor and prefrontal cortices c) the fronto-caudate tract (FCT), a fan-shaped tract, was documented to participate in connectivity of the prefrontal and premotor cortices to the head and body of the caudate nucleus and d) the cortico-tegmental tract(CTT) was invariably recorded to subserve the connectivity of the tegmental area with the fronto-parietal cortex. No hemispheric asymmetries were recorded for any of the implicated pathways. Sub-segmentation systems were also proposed for each of the aforementioned tracts. The structural connectivity and functional specialization of motor and premotor areas in the human brain remain vague to this day as most of the available evidence derives either from animal or tractographic studies. By using the fiber-microdissection technique as our main method of investigation, we provide sound structural evidence on the delicate anatomy of the related white matter pathways.

Keywords: neuroanatomy, premotor, motor, connectivity

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Authors: Amy Yee Kai Wan

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The arrival of a new baby to first-time parents is an exciting and joyous occasion, yet, the daunting task of raising the baby and the uncertainty of how it will affect the lives of the couple present a great challenge to them. This study examines the causes of conflicts and needs of the new parents through a qualitative research of five pairs of new parents in Hong Kong. Semi-structured in-depth qualitative interviews were conducted to explore the changes babies brought to their marriages, sources of support they received and found important and assistance they felt would help with their transition to parenthood. Thematic analysis was used to analyze the commonalities and differences between the five couples’ subjective experiences. Narrative analysis was used to compare the experiences of two parents who are the under-functioning parent of the couple, to study the different strategies they employed in response to the over-functioning parent and to analyze how the marital relationships were affected. Four main themes emerged from the study: 1) Change and adjustment in marital relationship, 2) parents’ level of involvement, 3) support in childcaring, and 4) challenges faced by the parents. Results from the study indicated that father involvement in childcaring is an important element in mother’s marital satisfaction Father’s marital satisfaction is dependent upon the mother – her satisfaction with father involvement, which affects the mother’s marital satisfaction. Marital convergence and co-parenting alliance acted as moderators for marital satisfaction. Implications from the study include: i) offering programmes that improve couple relationship and enhance parenting efficacy in tandem to improve overall marital satisfaction, and ii) offering prenatal counselling services or provide education to new parents from prenatal to postnatal period that can help couples reduce discrepancies between expectations and realities of their marital relationship and parenting responsibilities after their baby is born.

Keywords: co-parenting alliance, father involvement, marital convergence, maternal gatekeeping, new parents, transition to parenthood

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Authors: Abdulraaof H. Alqaili, Hamad A. Alsoliman

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Through progress in pavement design developments, a pavement design method was developed, which is titled the Mechanistic Empirical Pavement Design Guide (MEPDG). Nowadays, the evolution in roads network and highways is observed in Saudi Arabia as a result of increasing in traffic volume. Therefore, the MEPDG currently is implemented for flexible pavement design by the Saudi Ministry of Transportation. Implementation of MEPDG for local pavement design requires the calibration of distress models under the local conditions (traffic, climate, and materials). This paper aims to prepare data for calibration of MEPDG in Central Saudi Arabia. Thus, the first goal is data collection for the design of flexible pavement from the local conditions of the Riyadh region. Since, the modifying of collected data to input data is needed; the main goal of this paper is the analysis of collected data. The data analysis in this paper includes processing each: Trucks Classification, Traffic Growth Factor, Annual Average Daily Truck Traffic (AADTT), Monthly Adjustment Factors (MAFi), Vehicle Class Distribution (VCD), Truck Hourly Distribution Factors, Axle Load Distribution Factors (ALDF), Number of axle types (single, tandem, and tridem) per truck class, cloud cover percent, and road sections selected for the local calibration. Detailed descriptions of input parameters are explained in this paper, which leads to providing of an approach for successful implementation of MEPDG. Local calibration of MEPDG to the conditions of Riyadh region can be performed based on the findings in this paper.

Keywords: mechanistic-empirical pavement design guide (MEPDG), traffic characteristics, materials properties, climate, Riyadh

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Authors: Sara Vora (Hoxha)

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Defamation, invasion of privacy, and cyberbullying have all increased in tandem with the growth of the internet. European international private law may struggle to deal with such transgressions if they occur in many jurisdictions. The current study examines how effectively the legal system of European international private law addresses abuses of personality rights in cyberspace. The study starts by discussing how established legal frameworks are being threatened by online personality rights abuses. The article then looks into the rules and regulations of European international private law that are in place to handle overseas lawsuits. This article examines the different elements that courts evaluate when deciding which law to use in a particular case, focusing on the concepts of jurisdiction, choice of law, and recognition and execution of foreign judgements. Next, the research analyses the function of the European Union in preventing and punishing online personality rights abuses. Key pieces of law that control the collecting and processing of personal data on the Internet, including the General Data Protection Regulation (GDPR) and the e-Commerce Directive, are discussed. In addition, this article investigates how the ECtHR handles cases involving the infringement of personal freedoms, including privacy and speech. The article finishes with an assessment of how well the legal framework of European international private law protects individuals' right to privacy online. It draws attention to problems with the present legal structure, such as the inability to enforce international judgements, the inconsistency between national laws, and the necessity for stronger measures to safeguard people' rights online. This paper concludes that while European international private law provides a useful framework for dealing with violations of personality rights online, further harmonisation and stronger enforcement mechanisms are necessary to effectively protect individuals' rights in the digital age.

Keywords: European international private law, personality rights, internet, jurisdiction, cross-border disputes, data protection

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Authors: Sara D. Garduno-Diaz, Ramona Milcheva, Chanyu Xu

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Women’s reproductive stages (pre-pregnancy, pregnancy, and lactation) represent a time of higher micronutrient needs. With a healthy food selection as the first path of choice to cover these increased needs, tandem micronutrient supplementation is often required. Because pregnancy and lactation should be treated with care, all supplements consumed should be of quality ingredients and manufactured through controlled processes. This work describes the process followed for the development of plant-based multiple micronutrient supplements aimed at addressing the growing demand for natural ingredients of non-animal origin. A list of key nutrients for inclusion was prioritized, followed by the identification and selection of qualified raw ingredient providers. Nutrient absorption into the food matrix was carried out through natural processes. The outcome is a new line of products meeting the set criteria of being gluten and lactose-free, suitable for vegans/vegetarians, and without artificial conservatives. In addition, each product provides the consumer with 10 vitamins, 6 inorganic nutrients, 1 source of essential fatty acids, and 1 source of phytonutrients each (maca, moringa, and chlorella). Each raw material, as well as the final product, was submitted to microbiological control three-fold (in-house and external). The final micronutrient mix was then tested for human factor contamination, pesticides, total aerobic microbial count, total yeast count, and total mold count. The product was created with the aim of meeting product standards for the European Union, as well as specific requirements for the German market in the food and pharma fields. The results presented here reach the point of introduction of the newly developed product to the market, with acceptability and effectiveness results to be published at a later date.

Keywords: fertility, lactation, organic, pregnancy, vegetarian

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Authors: Lukman Ahmad Jamil, Heather M. Wallace

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Introduction: Numerous epidemiological studies have shown a positive as well as negative association and no association in some cases between chronic use of calcium channel blockers and the increased risk of developing cancer. However, these associations were enmeshed with controversies in the absence of laboratory based studies to back up those claims. Aim: The aim of this study was to determine in mechanistic terms the association between the long-term administration of nifedipine and diltiazem and increased risk of developing cancer using the human embryonic kidney (HEK293) cell line. Methods: Cell counting using the Trypan blue dye exclusion and 3-4, 5-Dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assays were used to investigate the effect of nifedipine and diltiazem on the growth pattern of HEK293 cells. Protein assay using modified Lowry method and analysis of intracellular polyamines concentration using Liquid Chromatography – Tandem Mass Spectrometry (LC-MS) were performed to ascertain the mechanism through which chronic use of nifedipine increases the risk of developing cancer. Results: Both nifedipine and diltiazem significantly increased the proliferation of HEK293 cells dose and time dependently. This proliferative effect after 24, 48 and 72-hour incubation period was observed at 0.78, 1.56 and 25 µM for nifedipine and 0.39, 1.56 and 25 µM for diltiazem, respectively. The increased proliferation of the cells was found to be statistically significantly (p<0.05). Furthermore, the increased proliferation of the cells induced by nifedipine was associated with the increase in the protein content and elevated intracellular polyamines concentration level. Conclusion: The chronic use of nifedipine is associated with increased proliferation of cells with concomitant elevation of polyamines concentration and elevated polyamine levels have been implicated in many malignant transformations and hence, these provide a possible explanation on the link between long term use of nifedipine and development of some human cancers. Further studies are needed to evaluate the cause of this association.

Keywords: cancer, nifedipine, polyamine, proliferation

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Authors: Abu Salim Mustafa

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Brucellosis is a zoonotic disease endemic in Kuwait. Human brucellosis can be caused by several Brucella species with Brucella melitensis causing the most severe and Brucella abortus the least severe disease. Furthermore, relapses are common after successful chemotherapy of patients. The classical biochemical methods of culture and serology for identification of Brucellae provide information about the species and serotypes only. However, to differentiate between relapse and reinfection/epidemiological investigations, the identification of genotypes using molecular methods is essential. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-16] were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. The 16S rRNA gene sequencing suggested that all the strains were B. melitensis and real-time PCR confirmed their species identity as B. melitensis. The ERIC-PCR band profiles produced a dendrogram of 75 branches suggesting each strain to be of a unique type. The cluster classification, based on ~ 80% similarity, divided all the ERIC genotypes into two clusters, A and B. Cluster A consisted of 9 ERIC genotypes (A1-A9) corresponding to 9 individual strains. Cluster B comprised of 13 ERIC genotypes (B1-B13) with B5 forming the largest cluster of 51 strains. MLVA-16 identified all isolates as B. melitensis and divided them into 71 MLVA-types. The cluster analysis of MLVA-16-types suggested that most of the strains in Kuwait originated from the East Mediterranean Region, a few from the African group and one new genotype closely matched with the West Mediterranean region. In conclusion, this work demonstrates that B. melitensis, the most pathogenic species of Brucella, is prevalent in Kuwait. Furthermore, MLVA-16 is the best molecular method, which can identify the Brucella species and genotypes as well as determine their origin in the global context. Supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: Brucella, ERIC-PCR, MLVA-16, RT-PCR, 16S rRNA gene sequencing

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Authors: Laure Wiest, Barbara Giroud, Azziz Assoumani, Francois Lestremau, Emmanuelle Vulliet

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Due to their production at high tonnages and their extensive use, surfactants are contaminants among those determined at the highest concentrations in wastewater. However, analytical methods and data regarding their occurrence in river water are scarce and concern only a few families, mainly anionic surfactants. The objective of this study was to develop an analytical method to extract and analyze a wide variety of surfactants in a minimum of steps, with a sensitivity compatible with the detection of ultra-traces in surface waters. 27 substances, from 12 families of surfactants, anionic, cationic and non-ionic were selected for method optimization. Different retention mechanisms for the extraction by solid phase extraction (SPE) were tested and compared in order to improve their detection by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The best results were finally obtained with a C18 grafted silica LC column and a polymer cartridge with hydrophilic lipophilic balance (HLB), and the method developed allows the extraction of the three types of surfactants with satisfactory recoveries. The final analytical method comprised only one extraction and two LC injections. It was validated and applied for the quantification of surfactants in 36 river samples. The method's limits of quantification (LQ), intra- and inter-day precision and accuracy were evaluated, and good performances were obtained for the 27 substances. As these compounds have many areas of application, contaminations of instrument and method blanks were observed and considered for the determination of LQ. Nevertheless, with LQ between 15 and 485 ng/L, and accuracy of over 80%, this method was suitable for monitoring surfactants in surface waters. Application on French river samples revealed the presence of anionic, cationic and non-ionic surfactants with median concentrations ranging from 24 ng/L for octylphenol ethoxylates (OPEO) to 4.6 µg/L for linear alkylbenzenesulfonates (LAS). The analytical method developed in this work will therefore be useful for future monitoring of surfactants in waters. Moreover, this method, which shows good performances for anionic, non-ionic and cationic surfactants, may be easily adapted to other surfactants.

Keywords: anionic surfactant, cationic surfactant, LC-MS/MS, non-ionic surfactant, SPE, surface water

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Authors: Dina Tawfic, Robert Hassan

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The current chapter addresses Al-Azhar's strategy to counter extremism in tandem with reflecting on the ideology of the Islamic establishment itself. The topic is motivated by the fact that some of the Western governments have been relying on Al-Azhar to counter the ideology of Islamist radicalism and violent extremism, in particular during the rise of the Islamic State in Syria and Iraq (known as ISIS/ ISIL/ Daesh) in 2014/2015. In his visit to Egypt in June 2016, Brett McGurk, the then U.S. envoy for the global coalition to counter ISIS, commended Al-Azhar’s “intellectual and reforming role” in refuting the ideology of extremism. On the other hand, Egyptian liberal intellectuals, such as Farag Fouda (1945- 1992) and Nasr Hamed Abu Zeid (1943-2010), had always questioned the ideological capability of Al-Azhar to counter extremism, citing the rigidity and resistance of the Islamic establishment to carry out genuine reformation. This chapter aims to discuss the following research questions: what is the strategy of Al-Azhar to counter extremism? Does Al-Azhar have a solid strategy to combat online propaganda produced by violent extremist groups? Is it applicable to identify Al-Azhar ideological identity? and is it capable of countering extremism? To answer these questions, I conducted intensive interviews with seven senior scholars and officials at Al-Azhar and the Endowments ministry from September to December 2020. Using a qualitative approach as a backdrop, this project uses semi-structured interviews to collect data. Participants were briefed on the purpose of the study and consented to be interviewed and to record their interviews. Some of the participants chose to conceal their names. All the interviews were conducted in Arabic via Zoom. The researcher then transcribed and translated the interviews into English. A purposive sample is used to select the seven interviewees, based on their prominence and experience in the field of counter-extremism and Al-Azhar affairs. The researcher uses a snowball sample to select the sample, in which a personal contact recommends other officials within the establishment.

Keywords: Al-Azhar, Egypt, Counter-Extremism, Political Islam, Ideology

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Authors: Qin Yang, Fan Zhang, Juan Du, Chongkui Sun, Krishna Kota, Yun-Ling Zheng

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Telomeres are specialized structures at chromosome ends consisting of tandem repetitive DNA sequences [(TTAGGG)n in humans] and associated proteins, which are necessary for telomere function. Telomere lengths are tightly regulated within a narrow range in normal human somatic cells, the basis of cellular senescence and aging. Previous studies have extensively focused on how short telomeres are extended and have demonstrated that telomerase plays a central role in telomere maintenance through elongating the short telomeres. However, the molecular mechanisms of regulating excessively long telomeres are unknown. Here, we found that telomerase enzymatic component hTERT plays a dual role in the regulation of telomeres length. We analyzed single telomere alterations at each chromosomal end led to the discoveries that hTERT shortens excessively long telomeres and elongates short telomeres simultaneously, thus maintaining the optimal telomere length at each chromosomal end for an efficient protection. The hTERT-mediated telomere shortening removes large segments of telomere DNA rapidly without inducing telomere dysfunction foci or affecting cell proliferation, thus it is mechanistically distinct from rapid telomere deletion. We found that expression of hTERT generates telomeric circular DNA, suggesting that telomere homologous recombination may be involved in this telomere shortening process. Moreover, the hTERT-mediated telomere shortening is required its enzymatic activity, but telomerase RNA component hTR is not involved in it. Furthermore, shelterin protein TPP1 interacts with hTERT and recruits it on telomeres to mediate telomere shortening. In addition, telomere-associated proteins, DKC1 and TCAB1 also play roles in this process. This novel hTERT-mediated telomere shortening mechanism not only exists in cancer cells, but also in primary human cells. Thus, the hTERT-mediated telomere shortening is expected to shift the paradigm on current molecular models of telomere length maintenance, with wide-reaching consequences in cancer and aging fields.

Keywords: aging, hTERT, telomerase, telomeres, human cells

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Authors: L. Kolatorova, J. Vitku, K. Adamcova, M. Simkova, M. Hill, A. Parizek, M. Duskova

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Endocrine disruptors (EDs) are substances leaching from various industrial products, which are able to interfere with the endocrine system. Their harmful effects on human health are generally well-known, and exposure during fetal development may have lasting effects. Fetal exposure and transplacental transport of bisphenol A (BPA) have been recently studied; however, less is known about alternatives such as bisphenol S (BPS), bisphenol F (BPF) and bisphenol AF (BPAF), which have started to appear in consumer products. The human organism is usually exposed to the mixture of EDs, out of which parabens are otherwise known to transfer placenta. The usage of many cosmetic, pharmaceutical and consumer products during the pregnancy that may contain parabens and bisphenols has led to the need for investigation. The aim of the study was to investigate the transplacental transport of BPA, its alternatives, and parabens, and to study their relation to fetal steroidogenesis. BPA, BPS, BPF, BPAF, methylparaben, ethylparaben, propylparaben, butylparaben, benzylparaben and 15 steroids including estrogens, corticoids, androgens and immunomodulatory ones were determined in 27 maternal (37th week of gestation) and cord plasma samples using liquid chromatography - tandem mass spectrometry methods. The statistical evaluation of the results showed significantly higher levels of BPA (p=0.0455) in cord plasma compared to maternal plasma. The results from multiple regression models investigated that in cord plasma, methylparaben, propylparaben and the sum of all measured parabens were inversely associated with testosterone levels. To our best knowledge, this study is the first attempt to determine the levels of alternative bisphenols in the maternal and cord blood, and also the first study reporting the simultaneous detection of bisphenols, parabens, and steroids in these biological fluids. Our study confirmed the transplacental transport of BPA, with likely accumulation in the fetal compartment. The negative association of cord blood parabens and testosterone levels highlights their possible risks, especially for the development of male fetuses. Acknowledgements: This work was supported by the project MH CR 17-30528 A from the Czech Health Research Council, MH CZ - DRO (Institute of Endocrinology - EÚ, 00023761) and by the MEYS CR (OP RDE, Excellent research - ENDO.CZ).

Keywords: bisphenol, endocrine disruptor, paraben, pregnancy, steroid

Procedia PDF Downloads 178