Search results for: transcription termination

Authors: Kyung Jin Kim, Jiwon Kwak, Jae-Hoon Lee, Soo Suk Lee

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MicroRNAs (miRNA) are a class of endogenous, single-stranded, small, and non-protein coding RNA molecules typically 20-25 nucleotides long. They are thought to regulate the expression of other genes in a broad range by binding to 3’- untranslated regions (3’-UTRs) of specific mRNAs. The detection of miRNAs is very important for understanding of the function of these molecules and in the diagnosis of variety of human diseases. However, detection of miRNAs is very challenging because of their short length and high sequence similarities within miRNA families. So, a simple-to-use, low-cost, and highly sensitive method for the detection of miRNAs is desirable. In this study, we demonstrate a novel bi-directional extension (BDE) assay. In the first step, a specific linear RT primer is hybridized to 6-10 base pairs from the 3’-end of a target miRNA molecule and then reverse transcribed to generate a cDNA strand. After reverse transcription, the cDNA was hybridized to the 3’-end which is BDE sequence; it played role as the PCR template. The PCR template was amplified in an SYBR green-based quantitative real-time PCR. To prove the concept, we used human brain total RNA. It could be detected quantitatively in the range of seven orders of magnitude with excellent linearity and reproducibility. To evaluate the performance of BDE assay, we contrasted sensitivity and specificity of the BDE assay against a commercially available poly (A) tailing method using miRNAs for let-7e extracted from A549 human epithelial lung cancer cells. The BDE assay displayed good performance compared with a poly (A) tailing method in terms of specificity and sensitivity; the CT values differed by 2.5 and the melting curve showed a sharper than poly (A) tailing methods. We have demonstrated an innovative, cost-effective BDE assay that allows improved sensitivity and specificity in detection of miRNAs. Dynamic range of the SYBR green-based RT-qPCR for miR-145 could be represented quantitatively over a range of 7 orders of magnitude from 0.1 pg to 1.0 μg of human brain total RNA. Finally, the BDE assay for detection of miRNA species such as let-7e shows good performance compared with a poly (A) tailing method in terms of specificity and sensitivity. Thus BDE proves a simple, low cost, and highly sensitive assay for various miRNAs and should provide significant contributions in research on miRNA biology and application of disease diagnostics with miRNAs as targets.

Keywords: bi-directional extension (BDE), microRNA (miRNA), poly (A) tailing assay, reverse transcription, RT-qPCR

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Authors: Hsiang-Ru Lin

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Cholesterol plays an essential role in the regulation of the progression of numerous important diseases including atherosclerosis and Alzheimer disease so the generation of suitable cholesterol-lowering reagents is urgent to develop. Liver X receptor (LXR) is a ligand-activated transcription factor whose natural ligands are cholesterols, oxysterols and glucose. Once being activated, LXR can transactivate the transcription action of various genes including CYP7A1, ABCA1, and SREBP1c, involved in the lipid metabolism, glucose metabolism and inflammatory pathway. Essentially, the upregulation of ABCA1 facilitates cholesterol efflux from the cells and attenuates the production of beta-amyloid (ABeta) 42 in brain so LXR is a promising target to develop the cholesterol-lowering reagents and preventative treatment of Alzheimer disease. Engelhardia roxburghiana is a deciduous tree growing in India, China, and Taiwan. However, its chemical composition is only reported to exhibit antitubercular and anti-inflammatory effects. In this study, four compounds, engelheptanoxides A, C, engelhardiol A, and B isolated from the root of Engelhardia roxburghiana were evaluated for their agonistic activity against LXR by the transient transfection reporter assays in the HepG2 cells. Furthermore, their interactive modes with LXR ligand binding pocket were generated by molecular modeling programs. By using the cell-based biological assays, engelheptanoxides A, C, engelhardiol A, and B showing no cytotoxic effect against the proliferation of HepG2 cells, exerted obvious LXR agonistic effects with similar activity as T0901317, a novel synthetic LXR agonist. Further modeling studies including docking and SAR (structure-activity relationship) showed that these compounds can locate in LXR ligand binding pocket in the similar manner as T0901317. Thus, LXR is one of nuclear receptors targeted by pharmaceutical industry for developing treatments of Alzheimer and atherosclerosis diseases. Importantly, the cell-based assays, together with molecular modeling studies suggesting a plausible binding mode, demonstrate that engelheptanoxides A, C, engelhardiol A, and B function as LXR agonists. This is the first report to demonstrate that the extract of Engelhardia roxburghiana contains LXR agonists. As such, these active components of Engelhardia roxburghiana or subsequent analogs may show important therapeutic effects through selective modulation of the LXR pathway.

Keywords: Liver X receptor (LXR), Engelhardia roxburghiana, CYP7A1, ABCA1, SREBP1c, HepG2 cells

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Authors: Said Boularouk, Didier Josselin, Eitan Altman

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In this paper, we present a vocal ontology of OpenStreetMap data for the apprehension of space by visually impaired people. Indeed, the platform based on produsage gives a freedom to data producers to choose the descriptors of geocoded locations. Unfortunately, this freedom, called also folksonomy leads to complicate subsequent searches of data. We try to solve this issue in a simple but usable method to extract data from OSM databases in order to send them to visually impaired people using Text To Speech technology. We focus on how to help people suffering from visual disability to plan their itinerary, to comprehend a map by querying computer and getting information about surrounding environment in a mono-modal human-computer dialogue.

Keywords: TTS, ontology, open street map, visually impaired

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Authors: Isabella Maurizio

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This research analyses the pronunciation of Hebrew consonants 'bgdkpt' in II- III C. E. in Palestine, through the confrontation of two kinds of data: the fragments of transliteration of Old Testament in the Greek alphabet, from the second column of Origen’s synopsis, called Hexapla, and Hebrew names transliterated in Greek documents, especially epigraphs. Origen is a very important author, not only for his bgdkpt theological and exegetic works: the Hexapla, synoptic six columns for a critical edition of Septuaginta, has a relevant role in attempting to reconstruct the pronunciation of Hebrew language before Masoretic punctuation. For this reason, at the beginning, it is important to analyze the column in order to study phonetic and linguistic phenomena. Among the most problematic data, there is the evidence from bgdkpt consonants, always represented as Greek aspirated graphemes. This transcription raised the question if their pronunciation was the only spirant, and consequently, the double one, that is, the stop/spirant contrast, was introduced by Masoretes. However, the phonetic and linguistic examination of the column alone is not enough to establish a real pronunciation of language: this paper is significant because a confrontation between the second column’s transliteration and Hebrew names found in Greek documents epigraphic ones mainly, is achieved. Palestine in II - III was a bilingual country: Greek and Aramaic language lived together, the first one like the official language, the second one as the principal mean of communication between people. For this reason, Hebrew names are often found in Greek documents of the same geographical area: a deep examination of bgdkpt’s transliteration can help to understand better which the real pronunciation of these consonants was, or at least it allows to evidence a phonetic tendency. As a consequence, the research considers the contemporary documents to Origen and the previous ones: the first ones testify a specific stadium of pronunciation, the second ones reflect phonemes’ evolution. Alexandrian documents are also examined: Origen was from there, and the influence of Greek language, spoken in his native country, must be considered. The epigraphs have another implication: they are totally free from morphological criteria, probably used by Origen in his column, because of their popular origin. Thus, a confrontation between the hexaplaric transliteration and Hebrew names is absolutely required, in Hexapla’s studies: first of all, it can be the second clue of a pronunciation already noted in the column; then because, for documents’ specific nature, it has more probabilities to be real, reflecting a daily use of language. The examination of data shows a general tendency to employ the aspirated graphemes for bgdkpt consonants’ transliteration. This probably means that they were closer to Greek aspirated consonants rather than to the plosive ones. The exceptions are linked to a particular status of the name, i.e. its history and origin. In this way, this paper gives its contribution to onomastic studies, too: indeed, the research may contribute to verify the diffusion and the treatment of Jewish names in Hellenized world and in the koinè language.

Keywords: bgdkpt consonants, Greek epigraphs, Jewish names, origen's Hexapla

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Authors: Lisa Mockli, Chelsea Sambells

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By combining the methodologies of history and translation studies, this study will explore the interplay between humanitarian action, politics, and translation within the advertising for a lesser-known Swiss child evacuation project of some 60.000 Belgium and French children to Switzerland for three month periods from 1940 to 1945. Inspired by Descriptive-Explanatory Translation Studies, this project compares Swiss speeches published between May and September 1942 (the termination of the evacuations). Radio broadcasts, leaflets and newspapers will triangulate the data. First, linguistic and content-related differences will be identified and described. Second, based on findings from the Swiss Federal Archives, the evidence from the comparative textual analysis will then be evaluated in order to explore how the speeches were modified, for what purpose, and which key issues were raised during their modification. By exploring these questions, this paper provides new insights into (I) Switzerland’s understanding of Swiss neutrality and humanitarianism during the Second World War, (II) the role of children in war and (III) the role of translation in shaping political discourse and humanitarian action. Moreover, this interdisciplinary approach also demonstrates how scholarly collaboration may help to make some elements of humanitarian action more self-reflexive and effective.

Keywords: children, history, humanitarianism, politics, translation

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Authors: Maureen Montalban

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The military is a predominantly male-dominated organisation that has entrenched hierarchical and patriarchal norms. Since 1975, women have been allowed to continue active service in the Australian Defence Force during pregnancy and after the birth of a child; prior to this time, pregnancy was grounds for automatic termination. The military and family, as institutions, make great demands on individuals with respect to their commitment, loyalty, time and energy. This research explores what it means to serve in the Australian Army as a woman through a gender lens, overlaid during a specific time period of their service; that is, during pregnancy, birth, and being a mother. It investigates the external demands faced by servicewomen who are mothers, whether it be from society, the Army, their teammates, their partners, or their children; and how they internally make sense of that with respect to their own identity and role as a mother, servicewoman, partner and as an individual. It also seeks to uncover how Australian Army servicewomen who are also mothers attempt to manage the dilemma of serving two greedy institutions when both expect and demand so much and whether this is, in fact, an impossible dilemma.

Keywords: women's health, gender studies, military culture, identity

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Authors: Seok Woo Chang, Kee Yeon Kum, Kwang Shik Bae, WooCheol Lee

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Aim: The objective of this study was to compare the biocompatibilities and mineralization potential of ProRoot MTA and newly developed calcium phosphate based cement, Capseal. Materials and Methods: The biocompatibilities and mineralization-related gene expressions (Bone sialoprotein (BSP) and osteocalcin (OCN)) of ProRoot MTA and Capseal were also compared by a methylthiazol tetrazolium (MTT) assay and reverse transcription-polymerization chain reaction (RT-PCR) analysis on 1, 3, and 7 days, respectively. Empty rings were used as control group. The results were statistically analyzed by Kruskal-Wallis test with a Bonferroni correction. P-value of < 0.05 was considered significant. Results: The biocompatibilities of ProRoot MTA and Capseal were equally favorable. ProRoot MTA and Capseal affected the messenger RNA expression of osteocalcin and osteonectin. Conclusions: Based on the results, both ProRoot MTA and Capseal could be a useful biomaterial in clinical endodontics.

Keywords: biocompatibility, calcium silicate cement, MTT, RT-PCR

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Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

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The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

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Authors: Kaoutar Ben Azzou, Hanaa Talei

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Human resources is an important department in each organization as it manages the life cycle of employees from recruitment training to retirement or termination of contracts. The recruitment process starts with a job opening, followed by a selection of the best-fit candidates from all applicants. Matching the best profile for a job position requires a manual way of looking at many CVs, which requires hours of work that can sometimes lead to choosing not the best profile. The work presented in this paper aims at reducing the workload of HR personnel by automating the preliminary stages of the candidate screening process, thereby fostering a more streamlined recruitment workflow. This tool introduces an automated system designed to help with the recruitment process by scanning candidates' CVs, extracting pertinent features, and employing machine learning algorithms to decide the most fitting job profile for each candidate. Our work employs natural language processing (NLP) techniques to identify and extract key features from unstructured text extracted from a CV, such as education, work experience, and skills. Subsequently, the system utilizes these features to match candidates with job profiles, leveraging the power of classification algorithms.

Keywords: automated recruitment, candidate screening, machine learning, human resources management

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Authors: Ayoub Kafyulilo, Petra Fisser, Joke Voogt

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This study was conducted to assess the continuation of the use of technology in science and mathematics teaching of the pre-service and in-service teachers who attended the professional development programme. It also assessed professional development, personal, institutional, and technological factors contributing to the continuous use of technology in teaching. The study involved 42 teachers, thirteen pre-service teachers, and twenty-nine in-service teachers. A mixed-method research approach was used to collect data for this study. Findings showed that the continuous use of technology in teaching after the termination of the professional development arrangement was high among the pre-service teachers, and differed for the in-service teachers. The regression model showed that knowledge and skills, access to technology and ease of use were strong predictors (R2 = 55.3%) of the teachers’ continuous use of technology after the professional development arrangement. The professional development factor did not have a direct effect on the continuous use of technology, rather had an influence on personal factors (knowledge and skills). In turn, the personal factors had influence on the institutional factors (access to technology) and technological factors (ease of use), which together had an effect on the teachers’ continuous use of technology in teaching.

Keywords: technology, professional development, teachers, science and mathematics

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Authors: Seok Woo Chang

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Aim: Capseal I and Capseal II are calcium silicate and calcium phosphate based experimental root canal sealer. The aim of this study was to evaluate the biocompatibility and mineralization potential of Capseal I and Capseal II. Materials and Methods: The biocompatibility and mineralization-related gene expression (alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN)) of Capseal I and Capseal II were compared using methylthiazol tetrazolium assay and reverse transcription-polymerization chain reaction analysis, respectively. The results were analyzed by Kruskal-Wallis test. P-value of < 0.05 was considered significant. Result: Both Capseal I and Capseal II were favorable in biocompatibility and influenced the messenger RNA expression of ALP and BSP. Conclusion: Within the limitation of this study, Capseal is biocompatible and have mineralization promoting potential, and thus could be a promising root canal sealer.

Keywords: biocompatibility, mineralization-related gene expression, Capseal I, Capseal II

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Authors: Ziba Omidi, Min-Ki Kim

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This research presents the design, fabrication and application of a flavor sensor for an integrated electronic tongue and electronic nose that can allow rapid characterization of multi-component mixtures in a solution. The odor gas and liquid are separated using hydrophobic porous membrane in micro fluidic channel. The sensor uses an array composed of microbeads in micromachined cavities localized on silicon wafer. Sensing occurs via colorimetric and fluorescence changes to receptors and indicator molecules that are attached to termination sites on the polymeric microbeads. As a result, the sensor array system enables simultaneous and near-real-time analyses using small samples and reagent volumes with the capacity to incorporate significant redundancies. One of the key parts of the system is a passive pump driven only by capillary force. The hydrophilic surface of the fluidic structure draws the sample into the sensor array without any moving mechanical parts. Since there is no moving mechanical component in the structure, the size of the fluidic structure can be compact and the fabrication becomes simple when compared to the device including active microfluidic components. These factors should make the proposed system inexpensive to mass-produce, portable and compatible with biomedical applications.

Keywords: optical sensor, semiconductor manufacturing, smell sensor, taste sensor

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Authors: Sonal Budhiraja, Biswabrata Pradhan

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This work considers statistical inference based on progressive Type-I interval censored data with random removal. The scheme of progressive Type-I interval censoring with random removal can be described as follows. Suppose n identical items are placed on a test at time T0 = 0 under k pre-fixed inspection times at pre-specified times T1 < T2 < . . . < Tk, where Tk is the scheduled termination time of the experiment. At inspection time Ti, Ri of the remaining surviving units Si, are randomly removed from the experiment. The removal follows a binomial distribution with parameters Si and pi for i = 1, . . . , k, with pk = 1. In this censoring scheme, the number of failures in different inspection intervals and the number of randomly removed items at pre-specified inspection times are observed. Asymptotic properties of the maximum likelihood estimators (MLEs) are established under some regularity conditions. A β-content γ-level tolerance interval (TI) is determined for two parameters Weibull lifetime model using the asymptotic properties of MLEs. The minimum sample size required to achieve the desired β-content γ-level TI is determined. The performance of the MLEs and TI is studied via simulation.

Keywords: asymptotic normality, consistency, regularity conditions, simulation study, tolerance interval

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Authors: Jian-Jun Shu

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The statistical properties of CRMs are explored by estimating similar-word set occurrence distribution. It is observed that CRMs tend to have a fat-tail distribution for similar-word set occurrence. Thus, the fat-tail test with two fatness coefficients is proposed to distinguish CRMs from non-CRMs, especially from exons. For the first fatness coefficient, the separation accuracy between CRMs and exons is increased as compared with the existing content-based CRM prediction method – fluffy-tail test. For the second fatness coefficient, the computing time is reduced as compared with fluffy-tail test, making it very suitable for long sequences and large data-base analysis in the post-genome time. Moreover, these indexes may be used to predict the CRMs which have not yet been observed experimentally. This can serve as a valuable filtering process for experiment.

Keywords: statistical approach, transcription factor binding sites, cis-regulatory modules, DNA sequences

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Authors: Kai Zhu, Shanhua He, Yujiao Chang

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This paper explores the virtual linguistic landscape of an emerging virtual language community in TikTok, a language community realizing immediate and non-immediate communication without a precise Spatio-temporal domain or a specific socio-cultural boundary or interpersonal network. This kind of language community generates a large number and various forms of virtual linguistic landscape, with which we conducted a virtual ethnographic survey together with telephone interviews to collect data from coping. We have been following two language communities in TikTok for several months so that we can illustrate the composition of the two language communities and some typical virtual language landscapes in both language communities first. Then we try to explore the reasons why and how they are formed through the organization, transcription, and analysis of the interviews. Our analysis reveals the richness and diversity of the virtual linguistic landscape, and finally, we summarize some of the characteristics of this language community.

Keywords: virtual linguistic landscape, virtual language community, virtual ethnographic survey, TikTok

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Authors: Mazen Al-Bulaihed, Nissar Wani, Abdulrahman Al-Arainy, Yasin Khan

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Partial Discharge (PD) measurements are widely used for diagnostic purposes in electrical equipment used in power systems. The main cause of these measurements is to prevent large power failures as cables are prone to aging, which usually results in embrittlement, cracking and eventual failure of the insulating and sheathing materials, exposing the conductor and risking a potential short circuit, a likely cause of the electrical fire. Many distribution networks rely heavily on medium voltage (MV) power cables. The presence of joints in these networks is a vital part of serving the consumer demand for electricity continuously. Such measurements become even more important when the extent of dependence increases. Moreover, it is known that the partial discharge in joints and termination are difficult to track and are the most crucial point of failures in large power systems. This paper discusses the diagnostic techniques of four samples of XLPE insulated cable joints, each included with a different type of defect. Experiments were carried out by measuring PD and tanδ at very low frequency applied high voltage. The results show the importance of combining PD and tanδ for effective cable assessment.

Keywords: partial discharge, tan delta, very low frequency, XLPE cable

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Authors: G. C. Santini, C. Potrich, L. Lunelli, L. Vanzetti, S. Marasso, M. Cocuzza, C. Pederzolli

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The relevance of circulating miRNAs as non-invasive biomarkers for several pathologies is nowadays undoubtedly clear, as they have been found to have both diagnostic and prognostic value able to add fundamental information to patients’ clinical picture. The availability of these data, however, relies on a time-consuming process spanning from the sample collection and processing to the data analysis. In light of this, strategies which are able to ease this procedure are in high demand and considerable effort have been made in developing Lab-on-a-chip (LOC) devices able to speed up and standardise the bench work. In this context, a very promising polydimethylsiloxane (PDMS)-based microdevice which integrates the processing of the biological sample, i.e. purification of extracellular miRNAs, and reverse transcription was previously developed in our lab. In this study, we aimed at the improvement of the miRNA extraction performances of this micro device by increasing the ability of its surface to absorb extracellular miRNAs from biological samples. For this purpose, we focused on the modulation of two properties of the material: roughness and charge. PDMS surface roughness was modulated by casting with several templates (terminated with silicon oxide coated by a thin anti-adhesion aluminum layer), followed by a panel of curing conditions. Atomic force microscopy (AFM) was employed to estimate changes at the nanometric scale. To introduce modifications in surface charge we functionalized PDMS with different mixes of positively charged 3-aminopropyltrimethoxysilanes (APTMS) and neutral poly(ethylene glycol) silane (PEG). The surface chemical composition was characterized by X-ray photoelectron spectroscopy (XPS) and the number of exposed primary amines was quantified with the reagent sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate (s-SDTB). As our final end point, the adsorption rate of all these different conditions was assessed by fluorescence microscopy by incubating a synthetic fluorescently-labeled miRNA. Our preliminary analysis identified casting on thermally grown silicon oxide, followed by a curing step at 85°C for 1 hour, as the most efficient technique to obtain a PDMS surface roughness in the nanometric scaleable to trap miRNA. In addition, functionalisation with 0.1% APTMS and 0.9% PEG was found to be a necessary step to significantly increase the amount of microRNA adsorbed on the surface, therefore, available for further steps as on-chip reverse transcription. These findings show a substantial improvement in the extraction efficiency of our PDMS microdevice, ultimately leading to an important step forward in the development of an innovative, easy-to-use and integrated system for the direct purification of less abundant circulating microRNAs.

Keywords: circulating miRNAs, diagnostics, Lab-on-a-chip, polydimethylsiloxane (PDMS)

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Authors: Mohammad Jamil Abd AlGhani

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The natural native antioxidants N,N-P-methyl phenyl acetone and N,N-phenyl acetone were isolated from the Iraqi crude oil region of Kirkuk by ion exchange and their structure was characterized by spectral and chemical analysis methods. Tetraline was used as a liquid hydrocarbon to detect the efficiency of isolated molecules at elevated temperature (393 K) that it has physicochemical specifications and structure closed to hydrocarbons fractionated from crude oil. The synthesized universal antioxidant 2,6-ditertiaryisobutyl-p-methyl phenol (Unol) with known stochiometric coefficient of inhibition equal to (2) was used as a model for comparative evaluation at the same conditions. Modified chemiluminescence method was used to find the amount of absorbed oxygen and the induction periods in and without the existence of isolated antioxidants molecules. The results of induction periods and quantity of absorbed oxygen during the oxidation process were measured by manometric installation. It was seen that at specific equal concentrations of N,N-phenyl acetone and N, N-P-methyl phenyl acetone in comparison with Unol at 393 K were with (2) and (2.5) times efficient than do Unol. It means that they had the ability to inhibit the formation of new free radicals and prevent the chain reaction to pass from the propagation to the termination step rather than decomposition of formed hydroperoxides.

Keywords: antioxidants, chemiluminescence, inhibition, Unol

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Authors: Kay Thinzar Phu, Lwin Lwin Oo

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In an intelligent transport system and advanced driver assistance system, the developing of real-time traffic sign detection and recognition (TSDR) system plays an important part in recent research field. There are many challenges for developing real-time TSDR system due to motion artifacts, variable lighting and weather conditions and situations of traffic signs. Researchers have already proposed various methods to minimize the challenges problem. The aim of the proposed research is to develop an efficient and effective TSDR in real time. This system proposes an adaptive thresholding method based on RGB color for traffic signs detection and new features for traffic signs recognition. In this system, the RGB color thresholding is used to detect the blue and yellow color traffic signs regions. The system performs the shape identify to decide whether the output candidate region is traffic sign or not. Lastly, new features such as termination points, bifurcation points, and 90’ angles are extracted from validated image. This system uses Myanmar Traffic Sign dataset.

Keywords: adaptive thresholding based on RGB color, blue color detection, feature extraction, yellow color detection

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Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

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Authors: Swati Srivastava, Mattia Lauriola, Yuval Gilad, Adi Kimchi, Yosef Yarden

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The glucocorticoid receptor (GR) has been proposed to play important, but incompletely understood roles in cancer. Glucocorticoids (GCs) are widely used as co-medication of various carcinomas, due to their ability to reduce the toxicity of chemotherapy. Furthermore, GR antagonism has proven to be a strategy to treat triple negative breast cancer and castration-resistant prostate cancer. These observations suggest differential GR involvement in cancer subtypes. The goal of our study has been to elaborate the current understanding of GR signaling in tumor progression and metastasis. Our study involves two cellular models, non-tumorigenic breast epithelial cells (MCF10A) and Ewing sarcoma cells (CHLA9). In our breast cell model, the results indicated that the GR agonist dexamethasone inhibits EGF-induced mammary cell migration, and this effect was blocked when cells were stimulated with a GR antagonist, namely RU486. Microarray analysis for gene expression revealed that the mechanism underlying inhibition involves dexamenthasone-mediated repression of well-known activators of EGFR signaling, alongside with enhancement of several EGFR’s negative feedback loops. Because GR mainly acts primarily through composite response elements (GREs), or via a tethering mechanism, our next aim has been to find the transcription factors (TFs) which can interact with GR in MCF10A cells.The TF-binding motif overrepresented at the promoter of dexamethasone-regulated genes was predicted by using bioinformatics. To validate the prediction, we performed high-throughput Protein Complementation Assays (PCA). For this, we utilized the Gaussia Luciferase PCA strategy, which enabled analysis of protein-protein interactions between GR and predicted TFs of mammary cells. A library comprising both nuclear receptors (estrogen receptor, mineralocorticoid receptor, GR) and TFs was fused to fragments of GLuc, namely GLuc(1)-X, X-GLuc(1), and X-GLuc(2), where GLuc(1) and GLuc(2) correspond to the N-terminal and C-terminal fragments of the luciferase gene.The resulting library was screened, in human embryonic kidney 293T (HEK293T) cells, for all possible interactions between nuclear receptors and TFs. By screening all of the combinations between TFs and nuclear receptors, we identified several positive interactions, which were strengthened in response to dexamethasone and abolished in response to RU486. Furthermore, the interactions between GR and the candidate TFs were validated by co-immunoprecipitation in MCF10A and in CHLA9 cells. Currently, the roles played by the uncovered interactions are being evaluated in various cellular processes, such as cellular proliferation, migration, and invasion. In conclusion, our assay provides an unbiased network analysis between nuclear receptors and other TFs, which can lead to important insights into transcriptional regulation by nuclear receptors in various diseases, in this case of cancer.

Keywords: epidermal growth factor, glucocorticoid receptor, protein complementation assay, transcription factor

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Authors: Ashish Jain, Satish Nayak

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Transdermal iontophoretic drug delivery system is viable drug delivery platform technology and has a strong market worldwide. Transdermal drug delivery system is particularly desirable for therapeutic agents that need prolonged administration at controlled plasma level. This makes appropriateness to antihypertensive and anti-diabetic agents for their transdermal development. Controlled zero order absorption, easily termination of drug delivery and easy to administration also support for popularity of transdermal delivery. In this current research iontophoretic delivery of various anti diabetic agents like glipizide, glibenclamide and glimepiride were carried out. The experiments were carried out at different drug concentrations and different current densities using cathodal iontophoresis. Diffusion cell for iontophoretic permeation study was modified according to Glikfield Design. Pig skin was used for in vitro permeation study and for the in-vivo study New Zealand rabbits were used. At all concentration level iontophoresis showed enhanced permeation rate compared to passive controls. Iontophoretic transports of selected drugs were found to be increased with the current densities. Results showed that target permeation rate for selected drugs could be achieved with the aid of iontophoresis by increasing the area in an appreciable range.

Keywords: transdermal, iontophoresis, pig skin, rabbits, glipizide, glibeclamide

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Authors: Najib Khumaidillah

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A conversation needs skills to create the good flow of it. The skills also need to be paid attention by a host like Oprah Winfrey and Justin Bieber as an artist. This study is aimed at describing turn taking strategies and adjacency pairs used by the speakers. The data are from one segment of The Oprah Winfrey Show’s transcription with Justin Bieber. Those are analyzed by Stenstorm’s turn taking theories and adjacency pairs theories. From the analysis, it was found that both speakers use various turn taking strategies and adjacency pairs. These findings are hoped to be an example for non-native English speaker in doing English conversation and advance people’s comprehension of how to organize good conversation structure.

Keywords: adjacency pairs, conversation structure, the Oprah Winfrey show, turn taking

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Authors: L. Kamandulytė-Merfeldienė

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The paper deals with the main issues of methodology of the Corpus of Spoken Lithuanian which was started to be developed in 2006. At present, the corpus consists of 300,000 grammatically annotated word forms. The creation of the corpus consists of three main stages: collecting the data, the transcription of the recorded data, and the grammatical annotation. Collecting the data was based on the principles of balance and naturality. The recorded speech was transcribed according to the CHAT requirements of CHILDES. The transcripts were double-checked and annotated grammatically using CHILDES. The development of the Corpus of Spoken Lithuanian has led to the constant increase in studies on spontaneous communication, and various papers have dealt with a distribution of parts of speech, use of different grammatical forms, variation of inflectional paradigms, distribution of fillers, syntactic functions of adjectives, the mean length of utterances.

Keywords: CHILDES, corpus of spoken Lithuanian, grammatical annotation, grammatical disambiguation, lexicon, Lithuanian

Procedia PDF Downloads 198

Authors: Qi Liu, Jiqin Yao, Xiaohu Zhou, Bo Zheng

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The first artificial cell was produced by Thomas Chang in the 1950s when he was trying to make a mimic of red blood cells. Since then, many different types of artificial cells have been constructed from one of the two approaches: a so-called bottom-up approach, which aims to create a cell from scratch, and a top-down approach, in which genes are sequentially knocked out from organisms until only the minimal genome required for sustaining life remains. In this project, bottom-up approach was used to build a new cell-free expression system which mimics artificial cell that capable of protein expression and communicate with each other. The artificial cells constructed from the bottom-up approach are usually lipid vesicles, polymersomes, hydrogels or aqueous droplets containing the nucleic acids and transcription-translation machinery. However, lipid vesicles based artificial cells capable of communication present several issues in the cell communication research: (1) The lipid vesicles normally lose the important functions such as protein expression within a few hours. (2) The lipid membrane allows the permeation of only small molecules and limits the types of molecules that can be sensed and released to the surrounding environment for chemical communication; (3) The lipid vesicles are prone to rupture due to the imbalance of the osmotic pressure. To address these issues, the hydrogel-based artificial cells were constructed in this work. To construct the artificial cell, polyacrylamide hydrogel was functionalized with Acrylate PEG Succinimidyl Carboxymethyl Ester (ACLT-PEG2000-SCM) moiety on the polymer backbone. The proteinaceous factors can then be immobilized on the polymer backbone by the reaction between primary amines of proteins and N-hydroxysuccinimide esters (NHS esters) of ACLT-PEG2000-SCM, the plasmid template and ribosome were encapsulated inside the hydrogel particles. Because the artificial cell could continuously express protein with the supply of nutrients and energy, the artificial cell-artificial cell communication and artificial cell-natural cell communication could be achieved by combining the artificial cell vector with designed plasmids. The plasmids were designed referring to the quorum sensing (QS) system of bacteria, which largely relied on cognate acyl-homoserine lactone (AHL) / transcription pairs. In one communication pair, “sender” is the artificial cell or natural cell that can produce AHL signal molecule by synthesizing the corresponding signal synthase that catalyzed the conversion of S-adenosyl-L-methionine (SAM) into AHL, while the “receiver” is the artificial cell or natural cell that can sense the quorum sensing signaling molecule form “sender” and in turn express the gene of interest. In the experiment, GFP was first immobilized inside the hydrogel particle to prove that the functionalized hydrogel particles could be used for protein binding. After that, the successful communication between artificial cell-artificial cell and artificial cell-natural cell was demonstrated, the successful signal between artificial cell-artificial cell or artificial cell-natural cell could be observed by recording the fluorescence signal increase. The hydrogel-based artificial cell designed in this work can help to study the complex communication system in bacteria, it can also be further developed for therapeutic applications.

Keywords: artificial cell, cell-free system, gene circuit, synthetic biology

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Authors: Narin Salehiyan, Ramin Ghasemi Shayan

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In order to investigate coronavirus RNA replication, transcription, recombination, protein processing and transport, virion assembly, the identification of coronavirus-specific cell receptors, and polymerase processing, the manipulation of coronavirus clones and complementary DNAs (cDNAs) of defective-interfering (DI) RNAs is the subject of this chapter. The idea of the Covid genome is nonsegmented, single-abandoned, and positive-sense RNA. When compared to other RNA viruses, its size is significantly greater, ranging from 27 to 32 kb. The quality encoding the enormous surface glycoprotein depends on 4.4 kb, encoding a forcing trimeric, profoundly glycosylated protein. This takes off exactly 20 nm over the virion envelope, giving the infection the appearance-with a little creative mind of a crown or coronet. Covid research has added to the comprehension of numerous parts of atomic science as a general rule, like the component of RNA union, translational control, and protein transport and handling. It stays a fortune equipped for creating startling experiences.

Keywords: covid-19, corona, virus, genome, genetic

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Authors: Lidija Simunovic

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A Close-out netting cause is a clause within master agreements which reduces credit risks. This clause contains the parties ' advance agreement that the occurrence of a certain event (such as the commencement of bankruptcy proceedings) will result in the termination of the contract and that their mutual claims will be calculated as a net lump-sum to be paid by one party to the other. The legal treatment of the enforceability of close-out netting clauses opens up many legal matters in comparative legal systems because it is not uniformly treated in comparative laws. Certain legal systems take a liberal approach and allow the enforcement of close-out netting clauses. Others are much stricter, and they limit or completely prohibit the enforcement of close-out netting clauses through the mandatory provisions of their national bankruptcy laws. The author analyzes the concept of close-out netting clauses in selected comparative legal systems and examines the differences in their legal treatment by using the historical, analytical, and comparative method. It results that special treatment of the close-out netting in national laws with a liberal approach is often forced by financial industry lobbies and introduced in national laws without the justified reasons. Contrary to that in legal systems with limited or prohibited approach on close-out netting the uncertain enforceability of the close-out netting clause causes potential credit risks. The detected discrepancy on the national legal treatment and national financial markets regarding close-out netting lead to the conclusion to author’s best knowledge that is not possible to use any national model of close-out netting as a role model which perfectly fits all.

Keywords: close-out netting clauses, derivatives, insolvency, offsetting

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Authors: Mark Berry

Abstract:

A number of studies have identified several factors involved in the malignant progression of cancer cells. The Primary modulator in driving inflammation to these transformed cells has been identified as the transcription factor known as nuclear factor-κB. This essential regulator of inflammation and the development of cancer, combined with a microenvironment of inflammation and signaling molecules, plays a major role in the malignant progression of cancer, and this progression is the result of the mutagenic predisposition of persistent substances that combat infection at tumor sites and other areas of chronic inflammation. Inflammation-induced tumors, and their inflammatory cells and regulators may be the primary source of metastasis of tumor cells through angiogenesis. Previous research on cytokines and chemokines, including their downstream targets, has been the focus of the cancer/inflammation connection. The identification of the biological mechanisms of other proteins vital to the inflammation cascade and their interactions are crucial to novel and effective therapeutic protocols for the treatment of inflammation-induced cancers. The Ancelim HSRP Protocol is just such a therapeutic intervention.

Keywords: ancelim, cancer, inflammation, tumor

Procedia PDF Downloads 505

Authors: Alaa M. Badr Eldin, Mayar M. Shahen, Mohammed S. Sedeek, Marwa I. Ezzat, Sawsan M. ElSonbaty, Muhammed A. Saad, Manal S. Afifi, Omar M. Sabry

Abstract:

Using HPLC-MS/MS technique, 133 polyphenolic compounds were identified in the methanol extract of pomegranate rind (Punica granatum L.). In-vitro cytotoxic activity against breast cancer cell line MCF-7 was investigated, with an IC50 of 54 ug/ml. In-silico molecular docking using ellagic acid, gallagic acid, and Punicalagin as model compounds identified in pomegranate rind extract confirmed the intriguing anti-estrogenic action of the key polyphenolic components in pomegranate rind extract. Surprisingly, taxol showed low activity compared to pomegranate compounds as ERα antagonist and ERβ agonist. Pomegranate rind extract enhanced apoptosis of breast cancer cells through upregulation of the caspase-3 expression and downregulation of NF-κB transcription factor.

Keywords: HPLC-MS/MS, pomegranate rind, cytotoxicity, MCF-7, ER, caspase-3, NF-kB

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Authors: Suresh Sharma, Neeti Goutam

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In India abortions are legal and with the exceedingly liberal and broadened law that was passed in 1971, “Medical Termination of Pregnancy Act” had opened a new window to Women’s’ freedom and choice over their fertility. This paper would like to focus on the factors responsible for or leading to unsafe abortion as well as such high incidence of abortion in India which can help in understanding the ways in which we can prevent this apathy. To study the intricacies involved in delivering safety to womanhood in terms of safe abortion practice which includes more trained personnel, detailed explanation and consequences of conducting an abortion, fine reporting, awareness regarding family planning measures and not only pressurizing them to sterilize immediately after an abortion but also prior to that informing them and lastly easy accessibility of Contraceptives with a educated and brief information on that. Data has been drawn from various sources such as National Family Household Survey (1, 2, 3), Health Management Information System and Annual Health Survey. To safeguard the interest of women when it comes to complications resulting from unsafe abortions, Reproductive Health laid its strict adherence to it in its guidelines. The Government could induce more measures in terms of family planning measures and increase in the number of skilled medical health force, chiefly in rural areas to prevent the illegality of abortions. But before that fine reporting on the number of abortions performed will give an insight to this very issue only then policies and programs will work much better in favor of women.

Keywords: abortion, MTP act, India, women

Procedia PDF Downloads 309