Search results for: switching sequences
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1020

Search results for: switching sequences

90 Leptospira Lipl32-Specific Antibodies: Therapeutic Property, Epitopes Characterization and Molecular Mechanisms of Neutralization

Authors: Santi Maneewatchararangsri, Wanpen Chaicumpa, Patcharin Saengjaruk, Urai Chaisri

Abstract:

Leptospirosis is a globally neglected disease that continues to be a significant public health and veterinary burden, with millions of cases reported each year. Early and accurate differential diagnosis of leptospirosis from other febrile illnesses and the development of a broad spectrum of leptospirosis vaccines are needed. The LipL32 outer membrane lipoprotein is a member of Leptospira adhesive matrices and has been found to exert hemolytic activity to erythrocytes in vitro. Therefore, LipL32 is regarded as a potential target for diagnosis, broad-spectrum leptospirosis vaccines, and for passive immunotherapy. In this study, we established LipL32-specific mouse monoclonal antibodies, mAbLPF1 and mAbLPF2, and their respective mouse- and humanized-engineered single chain variable fragment (ScFv). Their antibodies’ neutralizing activities against Leptospira-mediated hemolysis in vitro, and the therapeutic efficacy of mAbs against heterologous Leptospira infected hamsters were demonstrated. The epitope peptide of mAb LPF1 was mapped to a non-contiguous carboxy-terminal β-turn and amphipathic α-helix of LipL32 structure contributing to phospholipid/host cell adhesion and membrane insertion. We found that the mAbLPF2 epitope was located on the interacting loop of peptide binding groove of the LipL32 molecule responsible for interactions with host constituents. Epitope sequences are highly conserved among Leptospira spp. and are absent from the LipL32 superfamily of other microorganisms. Both epitopes are surface-exposed, readily accessible by mAbs, and immunogenic. However, they are less dominant when revealed by LipL32-specific immunoglobulins from leptospirosis-patient sera and rabbit hyperimmune serum raised by whole Leptospira. Our study also demonstrated an adhesion inhibitory activity of LipL32 protein to host membrane components and cells mediated by mAbs as well as an anti-hemolytic activity of the respective antibodies. The therapeutic antibodies, particularly the humanized-ScFv, have a potential for further development as non-drug therapeutic agent for human leptospirosis, especially in subjects allergic to antibiotics. The epitope peptides recognized by two therapeutic mAbs have potential use as tools for structure-function studies. Finally, protective peptides may be used as a target for epitope-based vaccines for control of leptospirosis.

Keywords: leptospira lipl32-specific antibodies, therapeutic epitopes, epitopes characterization, immunotherapy

Procedia PDF Downloads 276
89 Electret: A Solution of Partial Discharge in High Voltage Applications

Authors: Farhina Haque, Chanyeop Park

Abstract:

The high efficiency, high field, and high power density provided by wide bandgap (WBG) semiconductors and advanced power electronic converter (PEC) topologies enabled the dynamic control of power in medium to high voltage systems. Although WBG semiconductors outperform the conventional Silicon based devices in terms of voltage rating, switching speed, and efficiency, the increased voltage handling properties, high dv/dt, and compact device packaging increase local electric fields, which are the main causes of partial discharge (PD) in the advanced medium and high voltage applications. PD, which occurs actively in voids, triple points, and airgaps, is an inevitable dielectric challenge that causes insulation and device aging. The aging process accelerates over time and eventually leads to the complete failure of the applications. Hence, it is critical to mitigating PD. Sharp edges, airgaps, triple points, and bubbles are common defects that exist in any medium to high voltage device. The defects are created during the manufacturing processes of the devices and are prone to high-electric-field-induced PD due to the low permittivity and low breakdown strength of the gaseous medium filling the defects. A contemporary approach of mitigating PD by neutralizing electric fields in high power density applications is introduced in this study. To neutralize the locally enhanced electric fields that occur around the triple points, airgaps, sharp edges, and bubbles, electrets are developed and incorporated into high voltage applications. Electrets are electric fields emitting dielectric materials that are embedded with electrical charges on the surface and in bulk. In this study, electrets are fabricated by electrically charging polyvinylidene difluoride (PVDF) films based on the widely used triode corona discharge method. To investigate the PD mitigation performance of the fabricated electret films, a series of PD experiments are conducted on both the charged and uncharged PVDF films under square voltage stimuli that represent PWM waveform. In addition to the use of single layer electrets, multiple layers of electrets are also experimented with to mitigate PD caused by higher system voltages. The electret-based approach shows great promise in mitigating PD by neutralizing the local electric field. The results of the PD measurements suggest that the development of an ultimate solution to the decades-long dielectric challenge would be possible with further developments in the fabrication process of electrets.

Keywords: electrets, high power density, partial discharge, triode corona discharge

Procedia PDF Downloads 184
88 Enhancement of Cross-Linguistic Effect with the Increase in the Multilingual Proficiency during Early Childhood: A Case Study of English Language Acquisition by a Pre-School Child

Authors: Anupama Purohit

Abstract:

The paper is a study on the inevitable cross-linguistic effect found in the early multilingual learners. The cross-linguistic behaviour like code-mixing, code-switching, foreign accent, literal translation, redundancy and syntactic manipulation effected due to other languages on the English language output of a non-native pre-school child are discussed here. A case study method is adopted in this paper to support the claim of the title. A simultaneously tetra lingual pre-school child’s (within 1;3 to 4;0) language behaviour is analysed here. The sample output data of the child is gathered from the diary entries maintained by her family, regular observations and video recordings done since her birth. She is getting the input of her mother tongue, Sambalpuri, from her grandparents only; Hindi, the local language from her play-school and the neighbourhood; English only from her mother and occasional visit of other family friends; Odia only during the reading of the Odia story book. The child is exposed to code-mixing of all the languages throughout her childhood. But code-mixing, literal translation, redundancy and duplication were absent in her initial stage of multilingual acquisition. As the child was more proficient in English in comparison to her other first languages and had never heard code-mixing in English language; it was expected from her input pattern of English (one parent, English language) that she would maintain purity in her use of English while talking to the English language interlocutor. But with gradual increase in the language proficiency in each of the languages of the child, her handling of the multiple codes becomes deft cross-linguistically. It can be deduced from the case study that after attaining certain milestone proficiency in each language, the child’s linguistic faculty can operate at a metalinguistic level. The functional use of each morpheme, their arrangement in words and in the sentences, the supra segmental features, lexical-semantic mapping, culture specific use of a language and the pragmatic skills converge to give a typical childlike multilingual output in an intelligible manner to the multilingual people (with the same set of languages in combination). The result is appealing because for expressing the same ideas which the child used to speak (may be with grammatically wrong expressions) in one language, gradually, she starts showing cross-linguistic effect in her expressions. So the paper pleads for the separatist view from the very beginning of the holophrastic phase (as the child expresses in addressee-specific language); but development of a metalinguistic ability that helps the child in communicating in a sophisticated way according to the linguistic status of the addressee is unique to the multilingual child. This metalinguistic ability is independent of the mode if input of a multilingual child.

Keywords: code-mixing, cross-linguistic effect, early multilingualism, literal translation

Procedia PDF Downloads 279
87 Characterization of Mycoplasma Pneumoniae Causing Exacerbation of Asthma: A Prototypical Finding from Sri Lanka

Authors: Lakmini Wijesooriya, Vicki Chalker, Jessica Day, Priyantha Perera, N. P. Sunil-Chandra

Abstract:

M. pneumoniae has been identified as an etiology for exacerbation of asthma (EQA), although viruses play a major role in EOA. M. pneumoniae infection is treated empirically with macrolides, and its antibiotic sensitivity is not detected routinely. Characterization of the organism by genotyping and determination of macrolide resistance is important epidemiologically as it guides the empiric antibiotic treatment. To date, there is no such characterization of M. pneumoniae performed in Sri Lanka. The present study describes the characterization of M. pneumoniae detected from a child with EOA following a screening of 100 children with EOA. Of the hundred children with EOA, M. pneumoniae was identified only in one child by Real-Time polymerase chain reaction (PCR) test for identifying the community-acquired respiratory distress syndrome (CARDS) toxin nucleotide sequences. The M. pneumoniae identified from this patient underwent detection of macrolide resistance via conventional PCR, amplifying and sequencing the region of the 23S rDNA gene that contains single nucleotide polymorphisms that confer resistance. Genotyping of the isolate was performed via nested Multilocus Sequence Typing (MLST) in which eight (8) housekeeping genes (ppa, pgm, gyrB, gmk, glyA, atpA, arcC, and adk) were amplified via nested PCR followed by gene sequencing and analysis. As per MLST analysis, the M. pneumoniae was identified as sequence type 14 (ST14), and no mutations that confer resistance were detected. Resistance to macrolides in M. pneumoniae is an increasing problem globally. Establishing surveillance systems is the key to informing local prescriptions. In the absence of local surveillance data, antibiotics are started empirically. If the relevant microbiological samples are not obtained before antibiotic therapy, as in most occasions in children, the course of antibiotic is completed without a microbiological diagnosis. This happens more frequently in therapy for M. pneumoniae which is treated with a macrolide in most patients. Hence, it is important to understand the macrolide sensitivity of M. pneumoniae in the setting. The M. pneumoniae detected in the present study was macrolide sensitive. Further studies are needed to examine a larger dataset in Sri Lanka to determine macrolide resistance levels to inform the use of macrolides in children with EOA. The MLST type varies in different geographical settings, and it also provides a clue to the existence of macrolide resistance. The present study enhances the database of the global distribution of different genotypes of M. pneumoniae as this is the first such characterization performed with the increased number of samples to determine macrolide resistance level in Sri Lanka. M. pneumoniae detected from a child with exacerbation of asthma in Sri Lanka was characterized as ST14 by MLST and no mutations that confer resistance were detected.

Keywords: mycoplasma pneumoniae, Sri Lanka, characterization, macrolide resistance

Procedia PDF Downloads 164
86 Recognition of a Thinly Bedded Distal Turbidite: A Case Study from a Proterozoic Delta System, Chaossa Formation, Simla Group, Western Lesser Himalaya, India

Authors: Priyanka Mazumdar, Ananya Mukhopadhyay

Abstract:

A lot of progress has been achieved in the research of turbidites during the last decades. However, their relationship to delta systems still deserves further attention. This paper addresses example of fine grained turbidite from a pro-deltaic deposit of a Proterozoic mixed energy delta system exposed along Chaossa-Baliana river section of the Chaossa Formation of the Simla Basin. Lithostratigraphic analysis of the Chaossa Formation reveals three major facies associations (prodelta deposit-FA1, delta slope deposit-FA2 and delta front deposit-FA3) based on lithofacies types, petrography and sedimentary structures. Detailed process-based facies and paleoenvironmental analysis of the study area have led to identification of more than150 m thick coarsening-upwards deltaic successions composed of fine grained turbidites overlain by delta slope deposits. Erosional features are locally common at the base of turbidite beds and still more widespread at the top. The complete sequence has eight sub-divisions that are here termed T1 to T8. The basal subdivision (T1) comprises a massive graded unit with a sharp, scoured base, internal parallel-lamination and cross-lamination. The overlying sequence shows textural and compositional grading through alternating silt and mud laminae (T2). T2 is overlying by T3 which is characterized by climbing ripple and cross lamination. Parallel laminae are the predominant facies attributes of T4 which caps the T3 unit. T5 has a loaded scour base and is mainly characterized laminated silt. The topmost three divisions, graded mud (T6), ungraded mud (T7) and laminated mud (T8). The proposed sequence is analogous to the Bouma (1962) structural scheme for sandy turbidites. Repetition of partial sequences represents deposition from different stages of evolution of a large, muddy, turbidity flow. Detailed facies analysis of the study area reveals that the sediments of the turbidites developed during normal regression at the stage of stable or marginally rising sea level. Thin-bedded turbidites were deposited predominantly by turbidity currents in the relatively shallower part of the Simla basin. The fine-grained turbidites are developed by resedimentation of delta-front sands and slumping of upper pro-delta muds.

Keywords: turbidites, prodelta, proterozoic, Simla Basin, Bouma sequence

Procedia PDF Downloads 248
85 Systematic Identification of Noncoding Cancer Driver Somatic Mutations

Authors: Zohar Manber, Ran Elkon

Abstract:

Accumulation of somatic mutations (SMs) in the genome is a major driving force of cancer development. Most SMs in the tumor's genome are functionally neutral; however, some cause damage to critical processes and provide the tumor with a selective growth advantage (termed cancer driver mutations). Current research on functional significance of SMs is mainly focused on finding alterations in protein coding sequences. However, the exome comprises only 3% of the human genome, and thus, SMs in the noncoding genome significantly outnumber those that map to protein-coding regions. Although our understanding of noncoding driver SMs is very rudimentary, it is likely that disruption of regulatory elements in the genome is an important, yet largely underexplored mechanism by which somatic mutations contribute to cancer development. The expression of most human genes is controlled by multiple enhancers, and therefore, it is conceivable that regulatory SMs are distributed across different enhancers of the same target gene. Yet, to date, most statistical searches for regulatory SMs have considered each regulatory element individually, which may reduce statistical power. The first challenge in considering the cumulative activity of all the enhancers of a gene as a single unit is to map enhancers to their target promoters. Such mapping defines for each gene its set of regulating enhancers (termed "set of regulatory elements" (SRE)). Considering multiple enhancers of each gene as one unit holds great promise for enhancing the identification of driver regulatory SMs. However, the success of this approach is greatly dependent on the availability of comprehensive and accurate enhancer-promoter (E-P) maps. To date, the discovery of driver regulatory SMs has been hindered by insufficient sample sizes and statistical analyses that often considered each regulatory element separately. In this study, we analyzed more than 2,500 whole-genome sequence (WGS) samples provided by The Cancer Genome Atlas (TCGA) and The International Cancer Genome Consortium (ICGC) in order to identify such driver regulatory SMs. Our analyses took into account the combinatorial aspect of gene regulation by considering all the enhancers that control the same target gene as one unit, based on E-P maps from three genomics resources. The identification of candidate driver noncoding SMs is based on their recurrence. We searched for SREs of genes that are "hotspots" for SMs (that is, they accumulate SMs at a significantly elevated rate). To test the statistical significance of recurrence of SMs within a gene's SRE, we used both global and local background mutation rates. Using this approach, we detected - in seven different cancer types - numerous "hotspots" for SMs. To support the functional significance of these recurrent noncoding SMs, we further examined their association with the expression level of their target gene (using gene expression data provided by the ICGC and TCGA for samples that were also analyzed by WGS).

Keywords: cancer genomics, enhancers, noncoding genome, regulatory elements

Procedia PDF Downloads 90
84 Studying Second Language Learners' Language Behavior from Conversation Analysis Perspective

Authors: Yanyan Wang

Abstract:

This paper on second language teaching and learning uses conversation analysis (CA) approach and focuses on how second language learners of Chinese do repair when making clarification requests. In order to demonstrate their behavior in interaction, a comparison was made to study the differences between native speakers of Chinese with non-native speakers of Chinese. The significance of the research is to make second language teachers and learners aware of repair and how to seek clarification. Utilizing the methodology of CA, the research involved two sets of naturally occurring recordings, one of native speaker students and the other of non-native speaker students. Both sets of recording were telephone talks between students and teachers. There were 50 native speaker students and 50 non-native speaker students. From multiple listening to the recordings, the parts with repairs for clarification were selected for analysis which included the moments in the talk when students had problems in understanding or hearing the speaker and had to seek clarification. For example, ‘Sorry, I do not understand ‘and ‘Can you repeat the question? ‘were the parts as repair to make clarification requests. In the data, there were 43 such cases from native speaker students and 88 cases from non-native speaker students. The non-native speaker students were more likely to use repair to seek clarification. Analysis on how the students make clarification requests during their conversation was carried out by investigating how the students initiated problems and how the teachers repaired the problems. In CA term, it is called other-initiated self-repair (OISR), which refers to student-initiated teacher-repair in this research. The findings show that, in initiating repair, native speaker students pay more attention to mutual understanding (inter-subjectivity) while non-native speaker students, due to their lack of language proficiency, pay more attention to their status of knowledge (epistemic) switch. There are three major differences: 1, native Chinese students more often initiate closed-class OISR (seeking specific information in the request) such as repeating a word or phrases from the previous turn while non-native students more frequently initiate open-class OISR (not specifying clarification) such as ‘sorry, I don’t understand ‘. 2, native speakers’ clarification requests are treated by the teacher as understanding of the content while non-native learners’ clarification requests are treated by teacher as language proficiency problem. 3, native speakers don’t see repair as knowledge issue and there is no third position in the repair sequences to close repair while non-native learners take repair sequence as a time to adjust their knowledge. There is clear closing third position token such as ‘oh ‘ to close repair sequence so that the topic can go back. In conclusion, this paper uses conversation analysis approach to compare differences between native Chinese speakers and non-native Chinese learners in their ways of conducting repair when making clarification requests. The findings are useful in future Chinese language teaching and learning, especially in teaching pragmatics such as requests.

Keywords: conversation analysis (CA), clarification request, second language (L2), teaching implication

Procedia PDF Downloads 238
83 A Novel Chicken W Chromosome Specific Tandem Repeat

Authors: Alsu F. Saifitdinova, Alexey S. Komissarov, Svetlana A. Galkina, Elena I. Koshel, Maria M. Kulak, Stephen J. O'Brien, Elena R. Gaginskaya

Abstract:

The mystery of sex determination is one of the most ancient and still not solved until the end so far. In many species, sex determination is genetic and often accompanied by the presence of dimorphic sex chromosomes in the karyotype. Genomic sequencing gave the information about the gene content of sex chromosomes which allowed to reveal their origin from ordinary autosomes and to trace their evolutionary history. Female-specific W chromosome in birds as well as mammalian male-specific Y chromosome is characterized by the degeneration of gene content and the accumulation of repetitive DNA. Tandem repeats complicate the analysis of genomic data. Despite the best efforts chicken W chromosome assembly includes only 1.2 Mb from expected 55 Mb. Supplementing the information on the sex chromosome composition not only helps to complete the assembly of genomes but also moves us in the direction of understanding of the sex-determination systems evolution. A whole-genome survey to the assembly Gallus_gallus WASHUC 2.60 was applied for repeats search in assembled genome and performed search and assembly of high copy number repeats in unassembled reads of SRR867748 short reads datasets. For cytogenetic analysis conventional methods of fluorescent in situ hybridization was used for previously cloned W specific satellites and specifically designed directly labeled synthetic oligonucleotide DNA probe was used for bioinformatically identified repetitive sequence. Hybridization was performed with mitotic chicken chromosomes and manually isolated giant meiotic lampbrush chromosomes from growing oocytes. A novel chicken W specific satellite (GGAAA)n which is not co-localizes with any previously described classes of W specific repeats was identified and mapped with high resolution. In the composition of autosomes this repeat units was found as a part of upstream regions of gonad specific protein coding sequences. These findings may contribute to the understanding of the role of tandem repeats in sex specific differentiation regulation in birds and sex chromosome evolution. This work was supported by the postdoctoral fellowships from St. Petersburg State University (#1.50.1623.2013 and #1.50.1043.2014), the grant for Leading Scientific Schools (#3553.2014.4) and the grant from Russian foundation for basic researches (#15-04-05684). The equipment and software of Research Resource Center “Chromas” and Theodosius Dobzhansky Center for Genome Bioinformatics of Saint Petersburg State University were used.

Keywords: birds, lampbrush chromosomes, sex chromosomes, tandem repeats

Procedia PDF Downloads 368
82 Lattice Twinning and Detwinning Processes in Phase Transformation in Shape Memory Alloys

Authors: Osman Adiguzel

Abstract:

Shape memory effect is a peculiar property exhibited by certain alloy systems and based on martensitic transformation, and shape memory properties are closely related to the microstructures of the material. Shape memory effect is linked with martensitic transformation, which is a solid state phase transformation and occurs with the cooperative movement of atoms by means of lattice invariant shears on cooling from high-temperature parent phase. Lattice twinning and detwinning can be considered as elementary processes activated during the transformation. Thermally induced martensite occurs as martensite variants, in self-accommodating manner and consists of lattice twins. Also, this martensite is called the twinned martensite or multivariant martensite. Deformation of shape memory alloys in martensitic state proceeds through a martensite variant reorientation. The martensite variants turn into the reoriented single variants with deformation, and the reorientation process has great importance for the shape memory behavior. Copper based alloys exhibit this property in metastable β- phase region, which has DO3 –type ordered lattice in ternary case at high temperature, and these structures martensiticaly turn into the layered complex structures with lattice twinning mechanism, on cooling from high temperature parent phase region. The twinning occurs as martensite variants with lattice invariant shears in two opposite directions, <110 > -type directions on the {110}- type plane of austenite matrix. Lattice invariant shear is not uniform in copper based ternary alloys and gives rise to the formation of unusual layered structures, like 3R, 9R, or 18R depending on the stacking sequences on the close-packed planes of the ordered lattice. The unit cell and periodicity are completed through 18 atomic layers in case of 18R-structure. On the other hand, the deformed material recovers the original shape on heating above the austenite finish temperature. Meanwhile, the material returns to the twinned martensite structures (thermally induced martensite structure) in one way (irreversible) shape memory effect on cooling below the martensite finish temperature, whereas the material returns to the detwinned martensite structure (deformed martensite) in two-way (reversible) shape memory effect. Shortly one can say that the microstructural mechanisms, responsible for the shape memory effect are the twinning and detwinning processes as well as martensitic transformation. In the present contribution, x-ray diffraction, transmission electron microscopy (TEM) and differential scanning calorimetry (DSC) studies were carried out on two copper-based ternary alloys, CuZnAl, and CuAlMn.

Keywords: shape memory effect, martensitic transformation, twinning and detwinning, layered structures

Procedia PDF Downloads 415
81 X-Ray Detector Technology Optimization In CT Imaging

Authors: Aziz Ikhlef

Abstract:

Most of multi-slices CT scanners are built with detectors composed of scintillator - photodiodes arrays. The photodiodes arrays are mainly based on front-illuminated technology for detectors under 64 slices and on back-illuminated photodiode for systems of 64 slices or more. The designs based on back-illuminated photodiodes were being investigated for CT machines to overcome the challenge of the higher number of runs and connection required in front-illuminated diodes. In backlit diodes, the electronic noise has already been improved because of the reduction of the load capacitance due to the routing reduction. This translated by a better image quality in low signal application, improving low dose imaging in large patient population. With the fast development of multi-detector-rows CT (MDCT) scanners and the increasing number of examinations, the clinical community has raised significant concerns on radiation dose received by the patient in both medical and regulatory community. In order to reduce individual exposure and in response to the recommendations of the International Commission on Radiological Protection (ICRP) which suggests that all exposures should be kept as low as reasonably achievable (ALARA), every manufacturer is trying to implement strategies and solutions to optimize dose efficiency and image quality based on x-ray emission and scanning parameters. The added demands on the CT detector performance also comes from the increased utilization of spectral CT or dual-energy CT in which projection data of two different tube potentials are collected. One of the approaches utilizes a technology called fast-kVp switching in which the tube voltage is switched between 80kVp and 140kVp in fraction of a millisecond. To reduce the cross-contamination of signals, the scintillator based detector temporal response has to be extremely fast to minimize the residual signal from previous samples. In addition, this paper will present an overview of detector technologies and image chain improvement which have been investigated in the last few years to improve the signal-noise ratio and the dose efficiency CT scanners in regular examinations and in energy discrimination techniques. Several parameters of the image chain in general and in the detector technology contribute in the optimization of the final image quality. We will go through the properties of the post-patient collimation to improve the scatter-to-primary ratio, the scintillator material properties such as light output, afterglow, primary speed, crosstalk to improve the spectral imaging, the photodiode design characteristics and the data acquisition system (DAS) to optimize for crosstalk, noise and temporal/spatial resolution.

Keywords: computed tomography, X-ray detector, medical imaging, image quality, artifacts

Procedia PDF Downloads 242
80 Scenario of Some Minerals and Impact of Promoter Hypermethylation of DAP-K Gene in Gastric Carcinoma Patients of Kashmir Valley

Authors: Showkat Ahmad Bhat, Iqra Reyaz, Falaque ul Afshan, Ahmad Arif Reshi, Muneeb U. Rehman, Manzoor R. Mir, Sabhiya Majid, Sonallah, Sheikh Bilal, Ishraq Hussain

Abstract:

Background: Gastric cancer is the fourth most common cancer and the second leading cause of worldwide cancer-related deaths, with a wide variation in incidence rates across different geographical areas. The current view of cancer is that a malignancy arises from a transformation of the genetic material of a normal cell, followed by successive mutations and by chain of alterations in genes such as DNA repair genes, oncogenes, Tumor suppressor genes. Minerals are necessary for the functioning of several transcriptional factors, proteins that recognize certain DNA sequences and have been found to play a role in gastric cancer. Material Methods:The present work was a case control study and its aim was to ascertain the role of minerals and promoter hypermethylation of CpG islands of DAP-K gene in Gastric cancer patients among the Kashmiri population. Serum was extracted from all the samples and mineral estimation was done by AAS from serum, DNA was also extracted and was modified using bisulphite modification kit. Methylation-specific PCR was used for the analysis of the promoter hypermethylation status of DAP-K gene. The epigenetic analysis revealed that unlike other high risk regions, Kashmiri population has a different promoter hypermethylation profile of DAP-K gene and has different mineral profile. Results: In our study mean serum copper levels were significantly different for the two genders (p<0.05), while as no significant differences were observed for iron and zinc levels. In Methylation-specific PCR the methylation status of the promoter region of DAP-K gene was as 67.50% (27/40) of the gastric cancer tissues showed methylated DAP-K promoter and 32.50% (13/40) of the cases however showed unmethylated DAP-K promoter. Almost all 85% (17/20) of the histopathologically confirmed normal tissues showed unmethylated DAP-K promoter except only in 3 cases where DAP-K promoter was found to be methylated. The association of promoter hypermethylation with gastric cancer was evaluated by χ2 (Chi square) test and was found to be significant (P=0.0006). Occurrence of DAP-K methylation was found to be unequally distributed in males and females with more frequency in males than in females but the difference was not statistically significant (P =0.7635, Odds ratio=1.368 and 95% C.I=0.4197 to 4.456). When the frequency of DAP-K promoter methylation was compared with clinical staging of the disease, DAP-K promoter methylation was found to be certainly higher in Stage III/IV (85.71%) compared to Stage I/ II (57.69%) but the difference was not statistically significant (P =0.0673). These results suggest that DAP-K aberrant promoter hypermethylation in Kashmiri population contributes to the process of carcinogenesis in Gastric cancer and is reportedly one of the commonest epigenetic changes in the development of Gastric cancer.

Keywords: gastric cancer, minerals, AAS, hypermethylation, CpG islands, DAP-K gene

Procedia PDF Downloads 499
79 Facies Sedimentology and Astronomic Calibration of the Reinech Member (Lutetian)

Authors: Jihede Haj Messaoud, Hamdi Omar, Hela Fakhfakh Ben Jemia, Chokri Yaich

Abstract:

The Upper Lutetian alternating marl–limestone succession of Reineche Member was deposited over a warm shallow carbonate platform that permits Nummulites proliferation. High-resolution studies of 30 meters thick Nummulites-bearing Reineche Member, cropping out in Central Tunisia (Jebel Siouf), have been undertaken, regarding pronounced cyclical sedimentary sequences, in order to investigate the periodicity of cycles and their related orbital-scale oceanic and climatic changes. The palaeoenvironmental and palaeoclimatic data are preserved in several proxies obtainable through high-resolution sampling and laboratories measurement and analysis as magnetic susceptibility (MS) and carbonates contents in conjunction with a wireline logging tools. The time series analysis of proxies permits to establish cyclicity orders present in the studied intervals which could be linked to the orbital cycles. MS records provide high-resolution proxies for relative sea level change in Late Lutetian strata. The spectral analysis of MS fluctuations confirmed the orbital forcing by the presence of the complete suite of orbital frequencies in the precession of 23 ka, the obliquity of 41 ka, and notably the two modes of eccentricity of 100 and 405 ka. Regarding the two periodic sedimentary cycles detected by wavelet analysis of proxy fluctuations which coincide with the long-term 405 ka eccentricity cycle, the Reineche Member spanned 0,8 Myr. Wireline logging tools as gamma ray and sonic were used as a proxies to decipher cyclicity and trends in sedimentation and contribute to identifying and correlate units. There are used to constraint the highest frequency cyclicity modulated by a long term wavelength cycling apparently controlled by clay content. Interpreted as a result of variations in carbonate productivity, it has been suggested that the marl-limestone couplets, represent the sedimentary response to the orbital forcing. The calculation of cycle durations through Reineche Member, is used as a geochronometer and permit the astronomical calibration of the geologic time scale. Furthermore, MS coupled with carbonate contents, and fossil occurrences provide strong evidence for combined detrital inputs and marine surface carbonate productivity cycles. These two synchronous processes were driven by the precession index and ‘fingerprinted’ in the basic marl–limestone couplets, modulated by orbital eccentricity.

Keywords: magnetic susceptibility, cyclostratigraphy, orbital forcing, spectral analysis, Lutetian

Procedia PDF Downloads 278
78 X-Ray Detector Technology Optimization in Computed Tomography

Authors: Aziz Ikhlef

Abstract:

Most of multi-slices Computed Tomography (CT) scanners are built with detectors composed of scintillator - photodiodes arrays. The photodiodes arrays are mainly based on front-illuminated technology for detectors under 64 slices and on back-illuminated photodiode for systems of 64 slices or more. The designs based on back-illuminated photodiodes were being investigated for CT machines to overcome the challenge of the higher number of runs and connection required in front-illuminated diodes. In backlit diodes, the electronic noise has already been improved because of the reduction of the load capacitance due to the routing reduction. This is translated by a better image quality in low signal application, improving low dose imaging in large patient population. With the fast development of multi-detector-rows CT (MDCT) scanners and the increasing number of examinations, the clinical community has raised significant concerns on radiation dose received by the patient in both medical and regulatory community. In order to reduce individual exposure and in response to the recommendations of the International Commission on Radiological Protection (ICRP) which suggests that all exposures should be kept as low as reasonably achievable (ALARA), every manufacturer is trying to implement strategies and solutions to optimize dose efficiency and image quality based on x-ray emission and scanning parameters. The added demands on the CT detector performance also comes from the increased utilization of spectral CT or dual-energy CT in which projection data of two different tube potentials are collected. One of the approaches utilizes a technology called fast-kVp switching in which the tube voltage is switched between 80 kVp and 140 kVp in fraction of a millisecond. To reduce the cross-contamination of signals, the scintillator based detector temporal response has to be extremely fast to minimize the residual signal from previous samples. In addition, this paper will present an overview of detector technologies and image chain improvement which have been investigated in the last few years to improve the signal-noise ratio and the dose efficiency CT scanners in regular examinations and in energy discrimination techniques. Several parameters of the image chain in general and in the detector technology contribute in the optimization of the final image quality. We will go through the properties of the post-patient collimation to improve the scatter-to-primary ratio, the scintillator material properties such as light output, afterglow, primary speed, crosstalk to improve the spectral imaging, the photodiode design characteristics and the data acquisition system (DAS) to optimize for crosstalk, noise and temporal/spatial resolution.

Keywords: computed tomography, X-ray detector, medical imaging, image quality, artifacts

Procedia PDF Downloads 176
77 Modification of Escherichia coli PtolT Expression Vector via Site-Directed Mutagenesis

Authors: Yakup Ulusu, Numan Eczacıoğlu, İsa Gökçe, Helen Waller, Jeremy H. Lakey

Abstract:

Besides having the appropriate amino acid sequence to perform the function of proteins, it is important to have correct conformation after this sequence to process. To consist of this conformation depends on the amino acid sequence at the primary structure, hydrophobic interaction, chaperones and enzymes in charge of folding etc. Misfolded proteins are not functional and tend to be aggregated. Cysteine originating disulfide cross-links make stable this conformation of functional proteins. When two of the cysteine amino acids come side by side, disulfide bond is established that forms a cystine bridge. Due to this feature cysteine plays an important role on the formation of three-dimensional structure of many proteins. There are two cysteine amino acids (C44, C69) in the Tol-A-III protein. Unlike protein disulfide bonds from within his own, any non-specific cystine bridge causes a change in the three dimensional structure of the protein. Proteins can be expressed in various host cells as directly or fusion (chimeric). As a result of overproduction of the recombinant proteins, aggregation of insoluble proteins in the host cell can occur by forming a crystal structure called inclusion body. In general fusion proteins are produced for provide affinity tags to make proteins more soluble and production of some toxic proteins via fusion protein expression system like pTolT. Proteins can be modified by using a site-directed mutagenesis. By this way, creation of non-specific disulfide crosslinks can be prevented at fusion protein expression system via the present cysteine replaced by another amino acid such as serine, glycine or etc. To do this, we need; a DNA molecule that contains the gene that encodes for the target protein, required primers for mutation to be designed according to site directed mutagenesis reaction. This study was aimed to be replaced cysteine encoding codon TGT with serine encoding codon AGT. For this sense and reverse primers designed (given below) and used site-directed mutagenesis reaction. Several new copy of the template plasmid DNA has been formed with above mentioned mutagenic primers via polymerase chain reaction (PCR). PCR product consists of both the master template DNA (wild type) and the new DNA sequences containing mutations. Dpn-l endonuclease restriction enzyme which is specific for methylated DNA and cuts them to the elimination of the master template DNA. E. coli cells obtained after transformation were incubated LB medium with antibiotic. After purification of plasmid DNA from E. coli, the presence of the mutation was determined by DNA sequence analysis. Developed this new plasmid is called PtolT-δ.

Keywords: site directed mutagenesis, Escherichia coli, pTolT, protein expression

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76 Electrical Degradation of GaN-based p-channel HFETs Under Dynamic Electrical Stress

Authors: Xuerui Niu, Bolin Wang, Xinchuang Zhang, Xiaohua Ma, Bin Hou, Ling Yang

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The application of discrete GaN-based power switches requires the collaboration of silicon-based peripheral circuit structures. However, the packages and interconnection between the Si and GaN devices can introduce parasitic effects to the circuit, which has great impacts on GaN power transistors. GaN-based monolithic power integration technology is an emerging solution which can improve the stability of circuits and allow the GaN-based devices to achieve more functions. Complementary logic circuits consisting of GaN-based E-mode p-channel heterostructure field-effect transistors (p-HFETs) and E-mode n-channel HEMTs can be served as the gate drivers. E-mode p-HFETs with recessed gate have attracted increasing interest because of the low leakage current and large gate swing. However, they suffer from a poor interface between the gate dielectric and polarized nitride layers. The reliability of p-HFETs is analyzed and discussed in this work. In circuit applications, the inverter is always operated with dynamic gate voltage (VGS) rather than a constant VGS. Therefore, dynamic electrical stress has been simulated to resemble the operation conditions for E-mode p-HFETs. The dynamic electrical stress condition is as follows. VGS is a square waveform switching from -5 V to 0 V, VDS is fixed, and the source grounded. The frequency of the square waveform is 100kHz with the rising/falling time of 100 ns and duty ratio of 50%. The effective stress time is 1000s. A number of stress tests are carried out. The stress was briefly interrupted to measure the linear IDS-VGS, saturation IDS-VGS, As VGS switches from -5 V to 0 V and VDS = 0 V, devices are under negative-bias-instability (NBI) condition. Holes are trapped at the interface of oxide layer and GaN channel layer, which results in the reduction of VTH. The negative shift of VTH is serious at the first 10s and then changes slightly with the following stress time. However, different phenomenon is observed when VDS reduces to -5V. VTH shifts negatively during stress condition, and the variation in VTH increases with time, which is different from that when VDS is 0V. Two mechanisms exists in this condition. On the one hand, the electric field in the gate region is influenced by the drain voltage, so that the trapping behavior of holes in the gate region changes. The impact of the gate voltage is weakened. On the other hand, large drain voltage can induce the hot holes generation and lead to serious hot carrier stress (HCS) degradation with time. The poor-quality interface between the oxide layer and GaN channel layer at the gate region makes a major contribution to the high-density interface traps, which will greatly influence the reliability of devices. These results emphasize that the improved etching and pretreatment processes needs to be developed so that high-performance GaN complementary logics with enhanced stability can be achieved.

Keywords: GaN-based E-mode p-HFETs, dynamic electric stress, threshold voltage, monolithic power integration technology

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75 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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74 Recognition by the Voice and Speech Features of the Emotional State of Children by Adults and Automatically

Authors: Elena E. Lyakso, Olga V. Frolova, Yuri N. Matveev, Aleksey S. Grigorev, Alexander S. Nikolaev, Viktor A. Gorodnyi

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The study of the children’s emotional sphere depending on age and psychoneurological state is of great importance for the design of educational programs for children and their social adaptation. Atypical development may be accompanied by violations or specificities of the emotional sphere. To study characteristics of the emotional state reflection in the voice and speech features of children, the perceptual study with the participation of adults and the automatic recognition of speech were conducted. Speech of children with typical development (TD), with Down syndrome (DS), and with autism spectrum disorders (ASD) aged 6-12 years was recorded. To obtain emotional speech in children, model situations were created, including a dialogue between the child and the experimenter containing questions that can cause various emotional states in the child and playing with a standard set of toys. The questions and toys were selected, taking into account the child’s age, developmental characteristics, and speech skills. For the perceptual experiment by adults, test sequences containing speech material of 30 children: TD, DS, and ASD were created. The listeners were 100 adults (age 19.3 ± 2.3 years). The listeners were tasked with determining the children’s emotional state as “comfort – neutral – discomfort” while listening to the test material. Spectrographic analysis of speech signals was conducted. For automatic recognition of the emotional state, 6594 speech files containing speech material of children were prepared. Automatic recognition of three states, “comfort – neutral – discomfort,” was performed using automatically extracted from the set of acoustic features - the Geneva Minimalistic Acoustic Parameter Set (GeMAPS) and the extended Geneva Minimalistic Acoustic Parameter Set (eGeMAPS). The results showed that the emotional state is worse determined by the speech of TD children (comfort – 58% of correct answers, discomfort – 56%). Listeners better recognized discomfort in children with ASD and DS (78% of answers) than comfort (70% and 67%, respectively, for children with DS and ASD). The neutral state is better recognized by the speech of children with ASD (67%) than by the speech of children with DS (52%) and TD children (54%). According to the automatic recognition data using the acoustic feature set GeMAPSv01b, the accuracy of automatic recognition of emotional states for children with ASD is 0.687; children with DS – 0.725; TD children – 0.641. When using the acoustic feature set eGeMAPSv01b, the accuracy of automatic recognition of emotional states for children with ASD is 0.671; children with DS – 0.717; TD children – 0.631. The use of different models showed similar results, with better recognition of emotional states by the speech of children with DS than by the speech of children with ASD. The state of comfort is automatically determined better by the speech of TD children (precision – 0.546) and children with ASD (0.523), discomfort – children with DS (0.504). The data on the specificities of recognition by adults of the children’s emotional state by their speech may be used in recruitment for working with children with atypical development. Automatic recognition data can be used to create alternative communication systems and automatic human-computer interfaces for social-emotional learning. Acknowledgment: This work was financially supported by the Russian Science Foundation (project 18-18-00063).

Keywords: autism spectrum disorders, automatic recognition of speech, child’s emotional speech, Down syndrome, perceptual experiment

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73 Doctor-Patient Interaction in an L2: Pragmatic Study of a Nigerian Experience

Authors: Ayodele James Akinola

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This study investigated the use of English in doctor-patient interaction in a university teaching hospital from a southwestern state in Nigeria with the aim of identifying the role of communication in an L2, patterns of communication, discourse strategies, pragmatic acts, and contexts that shape the interaction. Jacob Mey’s Pragmatic Acts notion complemented with Emanuel and Emanuel’s model of doctor-patient relationship provided the theoretical standpoint. Data comprising 7 audio-recorded doctors-patient interactions were collected from a University Hospital in Oyo state, Nigeria. Interactions involving the use of English language were purposefully selected. These were supplemented with patients’ case notes and interviews conducted with doctors. Transcription was patterned alongside modified Arminen’s notations of conversation analysis. In the study, interaction in English between doctor and patients has the preponderance of direct-translation, code-mixing and switching, Nigerianism and use of cultural worldviews to express medical experience. Irrespective of these, three patterns communication, namely the paternalistic, interpretive, and deliberative were identified. These were exhibited through varying discourse strategies. The paternalistic model reflected slightly casual conversational conventions and registers. These were achieved through the pragmemic activities of situated speech acts, psychological and physical acts, via patients’ quarrel-induced acts, controlled and managed through doctors’ shared situation knowledge. All these produced empathising, pacifying, promising and instructing practs. The patients’ practs were explaining, provoking, associating and greeting in the paternalistic model. The informative model reveals the use of adjacency pairs, formal turn-taking, precise detailing, institutional talks and dialogic strategies. Through the activities of the speech, prosody and physical acts, the practs of declaring, alerting and informing were utilised by doctors, while the patients exploited adapting, requesting and selecting practs. The negotiating conversational strategy of the deliberative model featured in the speech, prosody and physical acts. In this model, practs of suggesting, teaching, persuading and convincing were utilised by the doctors. The patients deployed the practs of questioning, demanding, considering and deciding. The contextual variables revealed that other patterns (such as phatic and informative) are also used and they coalesced in the hospital within the situational and psychological contexts. However, the paternalistic model was predominantly employed by doctors with over six years in practice, while the interpretive, informative and deliberative models were found among registrar and others below six years of medical practice. Doctors’ experience, patients’ peculiarities and shared cultural knowledge influenced doctor-patient communication in the study.

Keywords: pragmatics, communication pattern, doctor-patient interaction, Nigerian hospital situation

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72 Analysis and Comparison of Asymmetric H-Bridge Multilevel Inverter Topologies

Authors: Manel Hammami, Gabriele Grandi

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In recent years, multilevel inverters have become more attractive for single-phase photovoltaic (PV) systems, due to their known advantages over conventional H-bridge pulse width-modulated (PWM) inverters. They offer improved output waveforms, smaller filter size, lower total harmonic distortion (THD), higher output voltages and others. The most common multilevel converter topologies, presented in literature, are the neutral-point-clamped (NPC), flying capacitor (FC) and Cascaded H-Bridge (CHB) converters. In both NPC and FC configurations, the number of components drastically increases with the number of levels what leads to complexity of the control strategy, high volume, and cost. Whereas, increasing the number of levels in case of the cascaded H-bridge configuration is a flexible solution. However, it needs isolated power sources for each stage, and it can be applied to PV systems only in case of PV sub-fields. In order to improve the ratio between the number of output voltage levels and the number of components, several hybrids and asymmetric topologies of multilevel inverters have been proposed in the literature such as the FC asymmetric H-bridge (FCAH) and the NPC asymmetric H-bridge (NPCAH) topologies. Another asymmetric multilevel inverter configuration that could have interesting applications is the cascaded asymmetric H-bridge (CAH), which is based on a modular half-bridge (two switches and one capacitor, also called level doubling network, LDN) cascaded to a full H-bridge in order to double the output voltage level. This solution has the same number of switches as the above mentioned AH configurations (i.e., six), and just one capacitor (as the FCAH). CAH is becoming popular, due to its simple, modular and reliable structure, and it can be considered as a retrofit which can be added in series to an existing H-Bridge configuration in order to double the output voltage levels. In this paper, an original and effective method for the analysis of the DC-link voltage ripple is given for single-phase asymmetric H-bridge multilevel inverters based on level doubling network (LDN). Different possible configurations of the asymmetric H-Bridge multilevel inverters have been considered and the analysis of input voltage and current are analytically determined and numerically verified by Matlab/Simulink for the case of cascaded asymmetric H-bridge multilevel inverters. A comparison between FCAH and the CAH configurations is done on the basis of the analysis of the DC and voltage ripple for the DC source (i.e., the PV system). The peak-to-peak DC and voltage ripple amplitudes are analytically calculated over the fundamental period as a function of the modulation index. On the basis of the maximum peak-to-peak values of low frequency and switching ripple voltage components, the DC capacitors can be designed. Reference is made to unity output power factor, as in case of most of the grid-connected PV generation systems. Simulation results will be presented in the full paper in order to prove the effectiveness of the proposed developments in all the operating conditions.

Keywords: asymmetric inverters, dc-link voltage, level doubling network, single-phase multilevel inverter

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71 Development of Loop Mediated Isothermal Amplification (Lamp) Assay for the Diagnosis of Ovine Theileriosis

Authors: Muhammad Fiaz Qamar, Uzma Mehreen, Muhammad Arfan Zaman, Kazim Ali

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Ovine Theileriosis is a world-wide concern, especially in tropical and subtropical areas, due to having tick abundance that has received less awareness in different developed and developing areas due to less worth of sheep, low to the middle level of infection in different small ruminants herd. Across Asia, the prevalence reports have been conducted to provide equivalent calculation of flock and animal level prevalence of Theileriosisin animals. It is a challenge for veterinarians to timely diagnosis & control of Theileriosis and famers because of the nature of the organism and inadequacy of restricted plans to control. All most work is based upon the development of such a technique which should be farmer-friendly, less expensive, and easy to perform into the field. By the timely diagnosis of this disease will decrease the irrational use of the drugs, and other plan was to determine the prevalence of Theileriosis in District Jhang by using the conventional method, PCR and qPCR, and LAMP. We quantify the molecular epidemiology of T.lestoquardiin sheep from Jhang districts, Punjab, Pakistan. In this study, we concluded that the overall prevalence of Theileriosis was (32/350*100= 9.1%) in sheep by using Giemsa staining technique, whereas (48/350*100= 13%) is observed by using PCR technique (56/350*100=16%) in qPCR and the LAMP technique have shown up to this much prevalence percentage (60/350*100= 17.1%). The specificity and sensitivity also calculated in comparison with the PCR and LAMP technique. Means more positive results have been shown when the diagnosis has been done with the help of LAMP. And there is little bit of difference between the positive results of PCR and qPCR, and the least positive animals was by using Giemsa staining technique/conventional method. If we talk about the specificity and sensitivity of the LAMP as compared to PCR, The cross tabulation shows that the results of sensitivity of LAMP counted was 94.4%, and specificity of LAMP counted was 78%. Advances in scientific field must be upon reality based ideas which can lessen the gaps and hurdles in the way of scientific research; the lamp is one of such techniques which have done wonders in adding value and helping human at large. It is such a great biological diagnostic tools and has helped a lot in the proper diagnosis and treatment of certain diseases. Other methods for diagnosis, such as culture techniques and serological techniques, have exposed humans with great danger. However, with the help of molecular diagnostic technique like LAMP, exposure to such pathogens is being avoided in the current era Most prompt and tentative diagnosis can be made using LAMP. Other techniques like PCR has many disadvantages when compared to LAMP as PCR is a relatively expensive, time consuming, and very complicated procedure while LAMP is relatively cheap, easy to perform, less time consuming, and more accurate. LAMP technique has removed hurdles in the way of scientific research and molecular diagnostics, making it approachable to poor and developing countries.

Keywords: distribution, thelaria, LAMP, primer sequences, PCR

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70 Teachers' and Learners' Experiences of Learners' Writing in English First Additional Language

Authors: Jane-Francis A. Abongdia, Thandiswa Mpiti

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There is an international concern to develop children’s literacy skills. In many parts of the world, the need to become fluent in a second language is essential for gaining meaningful access to education, the labour market and broader social functioning. In spite of these efforts, the problem still continues. The level of English language proficiency is far from satisfactory and these goals are unattainable by others. The issue is more complex in South Africa as learners are immersed in a second language (L2) curriculum. South Africa is a prime example of a country facing the dilemma of how to effectively equip a majority of its population with English as a second language or first additional language (FAL). Given the multilingual nature of South Africa with eleven official languages, and the position and power of English, the study investigates teachers’ and learners’ experiences on isiXhosa and Afrikaans background learners’ writing in English First Additional Language (EFAL). Moreover, possible causes of writing difficulties and teacher’s practices for writing are explored. The theoretical and conceptual framework for the study is provided by studies on constructivist theories and sociocultural theories. In exploring these issues, a qualitative approach through semi-structured interviews, classroom observations, and document analysis were adopted. This data is analysed by critical discourse analysis (CDA). The study identified a weak correlation between teachers’ beliefs and their actual teaching practices. Although the teachers believe that writing is as important as listening, speaking, reading, grammar and vocabulary, and that it needs regular practice, the data reveal that they fail to put their beliefs into practice. Moreover, the data revealed that learners were disturbed by their home language because when they do not know a word they would write either the isiXhosa or the Afrikaans equivalent. Code-switching seems to have instilled a sense of “dependence on translations” where some learners would not even try to answer English questions but would wait for the teacher to translate the questions into isiXhosa or Afrikaans before they could attempt to give answers. The findings of the study show a marked improvement in the writing performance of learners who used the process approach in writing. These findings demonstrate the need for assisting teachers to shift away from focusing only on learners’ performance (testing and grading) towards a stronger emphasis on the process of writing. The study concludes that the process approach to writing could enable teachers to focus on the various parts of the writing process which can give more freedom to learners to experiment their language proficiency. It would require that teachers develop a deeper understanding of the process/genre approaches to teaching writing advocated by CAPS. All in all, the study shows that both learners and teachers face numerous challenges relating to writing. This means that more work still needs to be done in this area. The present study argues that teachers teaching EFAL learners should approach writing as a critical and core aspect of learners’ education. Learners should be exposed to intensive writing activities throughout their school years.

Keywords: constructivism, English second language, language of learning and teaching, writing

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69 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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68 Genetically Modified Fuel-Ethanol Industrial Yeast Strains as Biocontrol Agents

Authors: Patrícia Branco, Catarina Prista, Helena Albergaria

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Industrial fuel-ethanol fermentations are carried out under non-sterile conditions, which favors the development of microbial contaminants, leading to huge economic losses. Wild yeasts such as Brettanomyces bruxellensis and lactic acid bacteria are the main contaminants of industrial bioethanol fermentation, affecting Saccharomyces cerevisiae performance and decreasing ethanol yields and productivity. In order to control microbial contaminations, the fuel-ethanol industry uses different treatments, including acid washing and antibiotics. However, these control measures carry environmental risks such as acid toxicity and the rise of antibiotic-resistant bacteria. Therefore, it is crucial to develop and apply less toxic and more environmentally friendly biocontrol methods. In the present study, an industrial fuel-ethanol starter, S. cerevisiae Ethanol-Red, was genetically modified to over-express AMPs with activity against fuel-ethanol microbial contaminants and evaluated regarding its biocontrol effect during mixed-culture alcoholic fermentations artificially contaminated with B. bruxellensis. To achieve this goal, S. cerevisiae Ethanol-Red strain was transformed with a plasmid containing the AMPs-codifying genes, i.e., partial sequences of TDH1 (925-963 bp) and TDH2/3 (925-963 bp) and a geneticin resistance marker. The biocontrol effect of those genetically modified strains was evaluated against B. bruxellensis and compared with the antagonistic effect exerted by the modified strain with an empty plasmid (without the AMPs-codifying genes) and the non-modified strain S. cerevisiae Ethanol-Red. For that purpose, mixed-culture alcoholic fermentations were performed in a synthetic must use the modified S. cerevisiae Ethanol-Red strains together with B. bruxellensis. Single-culture fermentations of B. bruxellensis strains were also performed as a negative control of the antagonistic effect exerted by S. cerevisiae strains. Results clearly showed an improved biocontrol effect of the genetically-modified strains against B. bruxellensis when compared with the modified Ethanol-Red strain with the empty plasmid (without the AMPs-codifying genes) and with the non-modified Ethanol-Red strain. In mixed-culture fermentation with the modified S. cerevisiae strain, B. bruxellensis culturability decreased from 5×104 CFU/mL on day-0 to less than 1 CFU/mL on day-10, while in single-culture B. bruxellensis increased its culturability from 6×104 to 1×106 CFU/mL in the first 6 days and kept this value until day-10. Besides, the modified Ethanol-Red strain exhibited an enhanced antagonistic effect against B. bruxellensis when compared with that induced by the non-modified Ethanol-Red strain. Indeed, culturability loss of B. bruxellensis after 10 days of fermentation with the modified Ethanol-Red strain was 98.7 and 100% higher than that occurred in fermentations performed with the non-modified Ethanol-Red and the empty-plasmid modified strain, respectively. Therefore, one can conclude that the S. cerevisiae genetically modified strain obtained in the present work may be a valuable solution for the mitigation of microbial contamination in fuel-ethanol fermentations, representing a much safer and environmentally friendly preservation strategy than the antimicrobial treatments (acid washing and antibiotics) currently applied in fuel-ethanol industry.

Keywords: antimicrobial peptides, fuel-ethanol microbial contaminations, fuel-ethanol fermentation, biocontrol agents, genetically-modified yeasts

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67 Closing the Loop between Building Sustainability and Stakeholder Engagement: Case Study of an Australian University

Authors: Karishma Kashyap, Subha D. Parida

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Rapid population growth and urbanization is creating pressure throughout the world. This has a dramatic effect on a lot of elements which include water, food, transportation, energy, infrastructure etc. as few of the key services. Built environment sector is growing concurrently to meet the needs of urbanization. Due to such large scale development of buildings, there is a need for them to be monitored and managed efficiently. Along with appropriate management, climate adaptation is highly crucial as well because buildings are one of the major sources of greenhouse gas emission in their operation phase. Buildings to be adaptive need to provide a triple bottom approach to sustainability i.e., being socially, environmentally and economically sustainable. Hence, in order to deliver these sustainability outcomes, there is a growing understanding and thrive towards switching to green buildings or renovating new ones as per green standards wherever possible. Academic institutions in particular have been following this trend globally. This is highly significant as universities usually have high occupancy rates because they manage a large building portfolio. Also, as universities accommodate the future generation of architects, policy makers etc., they have the potential of setting themselves as a best industry practice model for research and innovation for the rest to follow. Hence their climate adaptation, sustainable growth and performance management becomes highly crucial in order to provide the best services to users. With the objective of evaluating appropriate management mechanisms within academic institutions, a feasibility study was carried out in a recent 5-Star Green Star rated university building (housing the School of Construction) in Victoria (south-eastern state of Australia). The key aim was to understand the behavioral and social aspect of the building users, management and the impact of their relationship on overall building sustainability. A survey was used to understand the building occupant’s response and reactions in terms of their work environment and management. A report was generated based on the survey results complemented with utility and performance data which were then used to evaluate the management structure of the university. Followed by the report, interviews were scheduled with the facility and asset managers in order to understand the approach they use to manage the different buildings in their university campuses (old, new, refurbished), respective building and parameters incorporated in maintaining the Green Star performance. The results aimed at closing the communication and feedback loop within the respective institutions and assist the facility managers to deliver appropriate stakeholder engagement. For the wider design community, analysis of the data highlights the applicability and significance of prioritizing key stakeholders, integrating desired engagement policies within an institution’s management structures and frameworks and their effect on building performance

Keywords: building optimization, green building, post occupancy evaluation, stakeholder engagement

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66 Story of Per-: The Radial Network of One Lithuanian Prefix

Authors: Samanta Kietytė

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The object of this study is the verbal derivatives stemming from the Lithuanian prefix per-. The prefix under examination can be classified as prepositional, having descended from the preposition per, thereby sharing the same prototypical meaning – denoting movement OVER. These frequently co-occur within sentences (1). The aim of this paper is to conduct a semantic analysis of the prefix per- and to propose a possible radial network of its meanings. In essence, the aim is to identify the interrelationships existing between its meanings. 1) Jis peršoko per tvorą/ 3SG.NOM.M jump.PST.3 over fence.ACC.SG. /ʻHe jumped over the fenceʼ. The foundation of this work lies in the methodological and theoretical framework of cognitive linguistics. The prototypical meaning of prefixes consistently embodies spatial dimensions that can be described through image schemas. This entails the identification of the trajectory, the landmark, and the relation between them in the situation described by the prefixed verb. The meanings of linguistic units are not perceived as arbitrary, but rather, they are interconnected through semantic motivation. According to this perspective, a singular meaning within linguistic units is considered as prototypical, while additional meanings are descended (not necessarily directly) from it. For example, one of the per- meanings TRANSFER (2) is derived from the prototypical meaning OVER. 2) Prašau persiųsti vadovo laišką man./ Ask.PRS.1 forward.INF manager.GEN.SG email.ACC.SG 1.SG.DAT/ ʻPlease forward the manager‘s email to meʼ. Certain semantic relations are explained by the conceptual metaphor and metonymy theory. For instances, when prefixed verb has a meaning WIN (3) it is related to the prototypical meaning. In this case, the prefixed verb describes situations of winning in various ways. In the prototypical meaning, the trajector moves higher than the landmark, and winning is metaphorically perceived as being higher. 3) Sūnus peraugo tėvą./ Son.NOM.SG outgrow.PST.3 father.ACC.SG/ ʻThe son has outgrown the fatherʼ. The data utilized for this study was collected from the 2014 grammatically annotated text "Lithuanian Web (LithuanianWaC v2)", consisting of 63,645,700 words. Given that the corpus is grammatically lemmatized, the list of the 793 items was obtained using the wordlist function and specifying that verbs starting with per were searched. The list included not only prefixed verbs but also other verbs whose roots have the same letter sequences as prefixes. Also, words with misspellings, without diacritical marks, and words listed for lemmatization errors were rejected, and a total of 475 derivatives were left for further analysis. The semantic analysis revealed that there are 12 distinct meanings of the prefix per-. The spatial meanings were extracted by determining what a trajector is, what a landmark is, and what the relation between them is. The connection between non-spatial meanings and spatial ones occurs through semantic motivation established by identifying elements that correspond to the trajector and landmark. The analysis reveals that there are no strict boundaries among these meanings, instead showing a continuum that encompasses a central core and a peripheral association with their internal structure, i.e., some derivatives are more prototypical of a particular meaning than others.

Keywords: word-formation, cognitive semantics, metaphor, radial networks, prototype theory, prefix

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65 Transcription Skills and Written Composition in Chinese

Authors: Pui-sze Yeung, Connie Suk-han Ho, David Wai-ock Chan, Kevin Kien-hoa Chung

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Background: Recent findings have shown that transcription skills play a unique and significant role in Chinese word reading and spelling (i.e. word dictation), and written composition development. The interrelationships among component skills of transcription, word reading, word spelling, and written composition in Chinese have rarely been examined in the literature. Is the contribution of component skills of transcription to Chinese written composition mediated by word level skills (i.e., word reading and spelling)? Methods: The participants in the study were 249 Chinese children in Grade 1, Grade 3, and Grade 5 in Hong Kong. They were administered measures of general reasoning ability, orthographic knowledge, stroke sequence knowledge, word spelling, handwriting fluency, word reading, and Chinese narrative writing. Orthographic knowledge- orthographic knowledge was assessed by a task modeled after the lexical decision subtest of the Hong Kong Test of Specific Learning Difficulties in Reading and Writing (HKT-SpLD). Stroke sequence knowledge: The participants’ performance in producing legitimate stroke sequences was measured by a stroke sequence knowledge task. Handwriting fluency- Handwriting fluency was assessed by a task modeled after the Chinese Handwriting Speed Test. Word spelling: The stimuli of the word spelling task consist of fourteen two-character Chinese words. Word reading: The stimuli of the word reading task consist of 120 two-character Chinese words. Written composition: A narrative writing task was used to assess the participants’ text writing skills. Results: Analysis of covariance results showed that there were significant between-grade differences in the performance of word reading, word spelling, handwriting fluency, and written composition. Preliminary hierarchical multiple regression analysis results showed that orthographic knowledge, word spelling, and handwriting fluency were unique predictors of Chinese written composition even after controlling for age, IQ, and word reading. The interaction effects between grade and each of these three skills (orthographic knowledge, word spelling, and handwriting fluency) were not significant. Path analysis results showed that orthographic knowledge contributed to written composition both directly and indirectly through word spelling, while handwriting fluency contributed to written composition directly and indirectly through both word reading and spelling. Stroke sequence knowledge only contributed to written composition indirectly through word spelling. Conclusions: Preliminary hierarchical regression results were consistent with previous findings about the significant role of transcription skills in Chinese word reading, spelling and written composition development. The fact that orthographic knowledge contributed both directly and indirectly to written composition through word reading and spelling may reflect the impact of the script-sound-meaning convergence of Chinese characters on the composing process. The significant contribution of word spelling and handwriting fluency to Chinese written composition across elementary grades highlighted the difficulty in attaining automaticity of transcription skills in Chinese, which limits the working memory resources available for other composing processes.

Keywords: orthographic knowledge, transcription skills, word reading, writing

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64 Decentralized Peak-Shaving Strategies for Integrated Domestic Batteries

Authors: Corentin Jankowiak, Aggelos Zacharopoulos, Caterina Brandoni

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In a context of increasing stress put on the electricity network by the decarbonization of many sectors, energy storage is likely to be the key mitigating element, by acting as a buffer between production and demand. In particular, the highest potential for storage is when connected closer to the loads. Yet, low voltage storage struggles to penetrate the market at a large scale due to the novelty and complexity of the solution, and the competitive advantage of fossil fuel-based technologies regarding regulations. Strong and reliable numerical simulations are required to show the benefits of storage located near loads and promote its development. The present study was restrained from excluding aggregated control of storage: it is assumed that the storage units operate independently to one another without exchanging information – as is currently mostly the case. A computationally light battery model is presented in detail and validated by direct comparison with a domestic battery operating in real conditions. This model is then used to develop Peak-Shaving (PS) control strategies as it is the decentralized service from which beneficial impacts are most likely to emerge. The aggregation of flatter, peak- shaved consumption profiles is likely to lead to flatter and arbitraged profile at higher voltage layers. Furthermore, voltage fluctuations can be expected to decrease if spikes of individual consumption are reduced. The crucial part to achieve PS lies in the charging pattern: peaks depend on the switching on and off of appliances in the dwelling by the occupants and are therefore impossible to predict accurately. A performant PS strategy must, therefore, include a smart charge recovery algorithm that can ensure enough energy is present in the battery in case it is needed without generating new peaks by charging the unit. Three categories of PS algorithms are introduced in detail. First, using a constant threshold or power rate for charge recovery, followed by algorithms using the State Of Charge (SOC) as a decision variable. Finally, using a load forecast – of which the impact of the accuracy is discussed – to generate PS. A performance metrics was defined in order to quantitatively evaluate their operating regarding peak reduction, total energy consumption, and self-consumption of domestic photovoltaic generation. The algorithms were tested on load profiles with a 1-minute granularity over a 1-year period, and their performance was assessed regarding these metrics. The results show that constant charging threshold or power are far from optimal: a certain value is not likely to fit the variability of a residential profile. As could be expected, forecast-based algorithms show the highest performance. However, these depend on the accuracy of the forecast. On the other hand, SOC based algorithms also present satisfying performance, making them a strong alternative when the reliable forecast is not available.

Keywords: decentralised control, domestic integrated batteries, electricity network performance, peak-shaving algorithm

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63 Targeting Glucocorticoid Receptor Eliminate Dormant Chemoresistant Cancer Stem Cells in Glioblastoma

Authors: Aoxue Yang, Weili Tian, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Glucocorticoids (GCs) are used to treat GBM-associated edema. However, glucocorticoids participate in the physiological response to psychosocial stress, linked to poor cancer prognosis. This raises concern that glucocorticoids affect the tumor and BTSCs. Identifying markers specifically expressed by brain tumor stem cells (BTSCs) may enable specific therapies that spare their regular tissue-resident counterparts. By ribosome profiling analysis, we have identified that glycerol-3-phosphate dehydrogenase 1 (GPD1) is expressed by dormant BTSCs but not by NSCs. Through different stress-induced experiments in vitro, we found that only dexamethasone (DEXA) can significantly increase the expression of GPD1 in NSCs. Adversely, mifepristone (MIFE) which is classified as glucocorticoid receptors antagonists, could decrease GPD1 protein level and weaken the proliferation and stemness in BTSCs. Furthermore, DEXA can induce GPD1 expression in tumor-bearing mice brains and shorten animal survival, whereas MIFE has a distinct adverse effect that prolonged mice lifespan. Knocking out GR in NSC can block the upregulation of GPD1 inducing by DEXA, and we find the specific sequences on GPD1 promotor combined with GR, thus improving the efficiency of GPD1 transcription from CHIP-Seq. Moreover, GR and GPD1 are highly co-stained on GBM sections obtained from patients and mice. All these findings confirmed that GR could regulate GPD1 and loss of GPD1 Impairs Multiple Pathways Important for BTSCs Maintenance GPD1 is also a critical enzyme regulating glycolysis and lipid synthesis. We observed that DEXA and MIFE could change the metabolic profiles of BTSCs by regulating GPD1 to shift the transition of cell dormancy. Our transcriptome and lipidomics analysis demonstrated that cell cycle signaling and phosphoglycerides synthesis pathways contributed a lot to the inhibition of GPD1 caused by MIFE. In conclusion, our findings raise concern that treatment of GBM with GCs may compromise the efficacy of chemotherapy and contribute to BTSC dormancy. Inhibition of GR can dramatically reduce GPD1 and extend the survival duration of GBM-bearing mice. The molecular link between GPD1 and GR may give us an attractive therapeutic target for glioblastoma.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides

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62 The Future Control Rooms for Sustainable Power Systems: Current Landscape and Operational Challenges

Authors: Signe Svensson, Remy Rey, Anna-Lisa Osvalder, Henrik Artman, Lars Nordström

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The electric power system is undergoing significant changes. Thereby, the operation and control are becoming partly modified, more multifaceted and automated, and thereby supplementary operator skills might be required. This paper discusses developing operational challenges in future power system control rooms, posed by the evolving landscape of sustainable power systems, driven in turn by the shift towards electrification and renewable energy sources. A literature review followed by interviews and a comparison to other related domains with similar characteristics, a descriptive analysis was performed from a human factors perspective. Analysis is meant to identify trends, relationships, and challenges. A power control domain taxonomy includes a temporal domain (planning and real-time operation) and three operational domains within the power system (generation, switching and balancing). Within each operational domain, there are different control actions, either in the planning stage or in the real-time operation, that affect the overall operation of the power system. In addition to the temporal dimension, the control domains are divided in space between a multitude of different actors distributed across many different locations. A control room is a central location where different types of information are monitored and controlled, alarms are responded to, and deviations are handled by the control room operators. The operators’ competencies, teamwork skills, team shift patterns as well as control system designs are all important factors in ensuring efficient and safe electricity grid management. As the power system evolves with sustainable energy technologies, challenges are found. Questions are raised regarding whether the operators’ tacit knowledge, experience and operation skills of today are sufficient to make constructive decisions to solve modified and new control tasks, especially during disturbed operations or abnormalities. Which new skills need to be developed in planning and real-time operation to provide efficient generation and delivery of energy through the system? How should the user interfaces be developed to assist operators in processing the increasing amount of information? Are some skills at risk of being lost when the systems change? How should the physical environment and collaborations between different stakeholders within and outside the control room develop to support operator control? To conclude, the system change will provide many benefits related to electrification and renewable energy sources, but it is important to address the operators’ challenges with increasing complexity. The control tasks will be modified, and additional operator skills are needed to perform efficient and safe operations. Also, the whole human-technology-organization system needs to be considered, including the physical environment, the technical aids and the information systems, the operators’ physical and mental well-being, as well as the social and organizational systems.

Keywords: operator, process control, energy system, sustainability, future control room, skill

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61 Plant Microbiota of Coastal Halophyte Salicornia Ramossisima

Authors: Isabel N. Sierra-Garcia, Maria J. Ferreira, Sandro Figuereido, Newton Gomes, Helena Silva, Angela Cunha

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Plant-associated microbial communities are considered crucial in the adaptation of halophytes to coastal environments. The plant microbiota can be horizontally acquired from the environment or vertically transmitted from generation to generation via seeds. Recruiting of the microbial communities by the plant is affected by geographical location, soil source, host genotype, and cultivation practice. There is limited knowledge reported on the microbial communities in halophytes the influence of biotic and abiotic factors. In this work, the microbiota associated with the halophyte Salicornia ramosissima was investigated to determine whether the structure of bacterial communities is influenced by host genotype or soil source. For this purpose, two contrasting sites where S. ramosissima is established in the estuarine system of the Ria de Aveiro were investigated. One site corresponds to a natural salt marsh where S. ramosissima plants are present (wild plants), and the other site is a former salt pan that nowadays are subjected to intensive crop production of S. ramosissima (crop plants). Bacterial communities from the rhizosphere, seeds and root endosphere of S. ramossisima from both sites were investigated by sequencing bacterial 16S rRNA gene using the Illumina MiSeq platform. The analysis of the sequences showed that the three plant-associated compartments, rhizosphere, root endosphere, and seed endosphere, harbor distinct microbiomes. However, bacterial richness and diversity were higher in seeds of wild plants, followed by rhizosphere in both sites, while seeds in the crop site had the lowest diversity. Beta diversity measures indicated that bacterial communities in root endosphere and seeds were more similar in both wild and crop plants in contrast to rhizospheres that differed by local, indicating that the recruitment of the similar bacterial communities by the plant genotype is active in regard to the site. Moreover, bacterial communities from the root endosphere and rhizosphere were phylogenetically more similar in both sites, but the phylogenetic composition of seeds in wild and crop sites was distinct. These results indicate that cultivation practices affect the seed microbiome. However, minimal vertical transmission of bacteria from seeds to adult plants is expected. Seeds from the crop site showed higher abundances of Kushneria and Zunongwangia genera. Bacterial members of the classes Alphaprotebacteria and Bacteroidia were the most ubiquitous across sites and compartments and might encompass members of the core microbiome. These findings indicate that bacterial communities associated with S. ramosissima are more influenced by host genotype rather than local abiotic factors or cultivation practices. This study provides a better understanding of the composition of the plant microbiota in S. ramosissima , which is essential to predict the interactions between plant and associated microbial communities and their effects on plant health. This knowledge is useful to the manipulations of these microbial communities to enhance the health and productivity of this commercially important plant.

Keywords: halophytes, plant microbiome, Salicornia ramosissima, agriculture

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