Search results for: lncRNA profiling
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 255

Search results for: lncRNA profiling

225 Use of Pig as an Animal Model for Assessing the Differential MicroRNA Profiling in Kidney after Aristolochic Acid Intoxication

Authors: Daniela E. Marin, Cornelia Braicu, Gina C. Pistol, Roxana Cojocneanu-Petric, Ioana Berindan Neagoe, Mihail A. Gras, Ionelia Taranu

Abstract:

Aristolochic acid (AA) is a carcinogenic, mutagenic, and nephrotoxic compound commonly found in the Aristolochiaceae family of plants. AA is frequently associated with urothelial carcinoma of the upper urinary tract in human and animals and is considered as being responsible for Balkan Endemic Nephropathy. The pig provides a good animal model because the porcine urological system is very similar to that of humans, both in aspects of physiology and anatomy. MicroRNA (miRNA) are small non-coding RNAs that have an impact on a wide range of biological processes by regulating gene expression at post-transcriptional level. The objective of this study was to analyze the miRNA profiling in the kidneys of AA intoxicated swine. For this purpose, ten TOPIGS-40 crossbred weaned piglets, 4-week-old, male and females with an initial average body weight of 9.83 ± 0.5 kg were studied for 28 days. They were given ad libitum access to water and feed and randomly allotted to one of the following groups: control group (C) or aristolochic acid group (AA). They were fed a maize-soybean-meal-based diet contaminated or not with 0.25mgAA/kg. To profile miRNA in the kidneys of pigs, microarrays and bioinformatics approaches were applied to analyze the miRNA in the kidney of control and AA intoxicated pigs. After normalization, our results have shown that a total of 5 known miRNAs and 4 novel miRNAs had different profiling in the kidney of intoxicated animals versus control ones. Expression of miR-32-5p, miR-497-5p, miR-423-3p, miR-218-5p, miR-128-3p were up-regulated by 0.25mgAA/kg feed, while the expression of miR-9793-5p, miR-9835-3p, miR-9840-3p, miR-4334-5p was down-regulated. The microRNA profiling in kidney of intoxicated animals was associated with modified expression of target genes as: RICTOR, LASP1, SFRP2, DKK2, BMI1, RAF1, IGF1R, MAP2K1, WEE1, HDGF, BCL2, EIF4E etc, involved in cell division cycle, apoptosis, cell differentiation and cell migration, cell signaling, cancer etc. In conclusion, this study provides new data concerning the microRNA profiling in kidney after aristolochic acid intoxications with important implications for human and animal health.

Keywords: aristolochic acid, kidney, microRNA, swine

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224 Exploring the Profiles of Militants in the SWAT Valley of Pakistan

Authors: Lateef Hakim Zai Khyber, Syed Rashid Ali

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In the post 9/11 era, a new trend has developed of terrorist profiling on the basis of the ethnic, religious, political, psychological, social, and economic background of the terrorists to anticipate and assess the possible risk and to prevent and prosecute the suspected before they commit any violent act. The same profiling approach was adopted in different militant or terrorist de-radicalization and rehabilitation programs across the world in order to evaluate and identify the reasons and causes for joining terrorism in terms of push and pull factors. This paper attempts to explore and investigate the profiles of the detainees in the Sabaoon de-radicalization and Emancipation program, which aimed at de-radicalizing the former militants of Tehrik-e-Taliban (TTP) Pakistan in the Swat valley of Pakistan. This research attempted to use qualitative methods for collecting data, including a number of formal and informal open-ended interviews with the former staff members of Sabaoon to explore various aspects of the program, such as various approaches used at Sabaoon for terrorist profiling. It conducts a thorough examination of the profiles of the terrorist through their socioeconomic, ideological, emotional, intellectual, and psychological conditions and orientations, personal details, family issues, social preferences, etc. The study finds out that the majority of the terrorists belonged to the marginalized groups or lower class, including underprivileged tenants and poor laborers, of society having no access to land. They possess almost the same profiles, including low socioeconomic status, absence of a father or strict behavior of parents, large and combined families, lack of education, lack of religious understanding, etc. They also possess some common traits such as anxiety disorder, emotional instability, aggressive impulses and insecurity, depression, inferiority complex, lack of critical thinking and logical reasoning, authority-seeking behavior, and revenge-seeking behavior.

Keywords: terrorist profiling, Sabaoon, de-radicalization, rehabilitation, Swat, Pakistan, juvenile militants

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223 Business Intelligence for Profiling of Telecommunication Customer

Authors: Rokhmatul Insani, Hira Laksmiwati Soemitro

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Business Intelligence is a methodology that exploits the data to produce information and knowledge systematically, business intelligence can support the decision-making process. Some methods in business intelligence are data warehouse and data mining. A data warehouse can store historical data from transactional data. For data modelling in data warehouse, we apply dimensional modelling by Kimball. While data mining is used to extracting patterns from the data and get insight from the data. Data mining has many techniques, one of which is segmentation. For profiling of telecommunication customer, we use customer segmentation according to customer’s usage of services, customer invoice and customer payment. Customers can be grouped according to their characteristics and can be identified the profitable customers. We apply K-Means Clustering Algorithm for segmentation. The input variable for that algorithm we use RFM (Recency, Frequency and Monetary) model. All process in data mining, we use tools IBM SPSS modeller.

Keywords: business intelligence, customer segmentation, data warehouse, data mining

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222 HPTLC Metabolite Fingerprinting of Artocarpus champeden Stembark from Several Different Locations in Indonesia and Correlation with Antimalarial Activity

Authors: Imam Taufik, Hilkatul Ilmi, Puryani, Mochammad Yuwono, Aty Widyawaruyanti

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Artocarpus champeden Spreng stembark (Moraceae) in Indonesia well known as ‘cempedak’ had been traditionally used for malarial remedies. The difference of growth locations could cause the difference of metabolite profiling. As a consequence, there were difference antimalarial activities in spite of the same plants. The aim of this research was to obtain the profile of metabolites that contained in A. champeden stembark from different locations in Indonesia for authentication and quality control purpose of this extract. The profiling had been performed by HPTLC-Densitometry technique and antimalarial activity had been also determined by HRP2-ELISA technique. The correlation between metabolite fingerprinting and antimalarial activity had been analyzed by Principle Component Analysis, Hierarchical Clustering Analysis and Partial Least Square. As a result, there is correlation between the difference metabolite fingerprinting and antimalarial activity from several different growth locations.

Keywords: antimalarial, artocarpus champeden spreng, metabolite fingerprinting, multivariate analysis

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221 Gut Metabolite Profiling of the Ethnic Groups from Assam, India

Authors: Madhusmita Dehingia, Supriyo Sen, Bhuwan Bhaskar, Tulsi Joishy, Mojibur R. Khan

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Human gut microbes and their metabolites are important for maintaining homeostasis in the gut and are responsible for many metabolic and immune mediated diseases. In the present study, we determined the profiles of the gut metabolites of five different ethnic groups (Bodo, Tai-Phake, Karbi, Tea tribe and Tai-Aiton) of Assam. Fecal metabolite profiling of the 39 individuals belonging to the ethnic groups was carried out using Gas chromatography – Mass spectrometry (GC-MS), and comparison was performed among the tribes for common and unique metabolites produced within their gut. Partial Least Squares Discriminant Analysis (PLS-DA) of the metabolites suggested that the individuals grouped according to their ethnicity. Among the 66 abundant metabolites, 12 metabolites were found to be common among the five ethnic groups. Additionally, ethnicity wise some unique metabolites were also detected. For example, the tea tribe of Assam contained the tea components, Aniline and Benzoate more in their gut in comparison to others. Metabolites of microbial origin were also correlated with the already published metagenomic data of the same ethnic group and functional analysis were carried out based on human metabolome database.

Keywords: ethnicity, gut microbiota, GC-MS, metabolites

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220 Approach to Honey Volatiles' Profiling by Gas Chromatography and Mass Spectrometry

Authors: Igor Jerkovic

Abstract:

Biodiversity of flora provides many different nectar sources for the bees. Unifloral honeys possess distinctive flavours, mainly derived from their nectar sources (characteristic volatile organic components (VOCs)). Specific or nonspecific VOCs (chemical markers) could be used for unifloral honey characterisation as addition to the melissopalynologycal analysis. The main honey volatiles belong, in general, to three principal categories: terpenes, norisoprenoids, and benzene derivatives. Some of these substances have been described as characteristics of the floral source, and other compounds, like several alcohols, branched aldehydes, and furan derivatives, may be related to the microbial purity of honey processing and storage conditions. Selection of the extraction method for the honey volatiles profiling should consider that heating of the honey produce different artefacts and therefore conventional methods of VOCs isolation (such as hydrodistillation) cannot be applied for the honey. Two-way approach for the isolation of the honey VOCs was applied using headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE). The extracts were analysed by gas chromatography and mass spectrometry (GC-MS). HS-SPME (with the fibers of different polarity such as polydimethylsiloxane/ divinylbenzene (PDMS/DVB) or divinylbenzene/carboxene/ polydimethylsiloxane (DVB/CAR/PDMS)) enabled isolation of high volatile headspace VOCs of the honey samples. Among them, some characteristic or specific compounds can be found such as 3,4-dihydro-3-oxoedulan (in Centaurea cyanus L. honey) or 1H-indole, methyl anthranilate, and cis-jasmone (in Citrus unshiu Marc. honey). USE with different solvents (mainly dichloromethane or the mixture pentane : diethyl ether 1 : 2 v/v) enabled isolation of less volatile and semi-volatile VOCs of the honey samples. Characteristic compounds from C. unshiu honey extracts were caffeine, 1H-indole, 1,3-dihydro-2H-indol-2-one, methyl anthranilate, and phenylacetonitrile. Sometimes, the selection of solvent sequence was useful for more complete profiling such as sequence I: pentane → diethyl ether or sequence II: pentane → pentane/diethyl ether (1:2, v/v) → dichloromethane). The extracts with diethyl ether contained hydroquinone and 4-hydroxybenzoic acid as the major compounds, while (E)-4-(r-1’,t-2’,c-4’-trihydroxy-2’,6’,6’-trimethylcyclo-hexyl)but-3-en-2-one predominated in dichloromethane extracts of Allium ursinum L. honey. With this two-way approach, it was possible to obtain a more detailed insight into the honey volatile and semi-volatile compounds and to minimize the risks of compound discrimination due to their partial extraction that is of significant importance for the complete honey profiling and identification of the chemical biomarkers that can complement the pollen analysis.

Keywords: honey chemical biomarkers, honey volatile compounds profiling, headspace solid-phase microextraction (HS-SPME), ultrasonic solvent extraction (USE)

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219 Metabolic and Phylogenetic Profiling of Rhizobium leguminosarum Strains Isolated from NZ Soils of Varying pH

Authors: Anish Shah, Steve A. Wakelin, Derrick Moot, Aurélie Laugraud, Hayley J. Ridgway

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A mixed pasture system of ryegrass-clover is used in New Zealand, where clovers are generally inoculated with commercially available strains of rhizobia. The community of rhizobia living in the soil and the way in which they interact with the plant are affected by different biotic and abiotic factors. In general, bacterial richness and diversity in soil varies by soil pH. pH also affects cell physiology and acts as a master variable that controls the wider soil physiochemical conditions such as P availability, Al release and micronutrient availability. As such, pH can have both primary and secondary effects on soil biology and processes. The aim of this work was to investigate the effect of soil pH on the genetic diversity and metabolic profile of Rhizobium leguminosarum strains nodulating clover. Soils were collected from 12 farms across New Zealand which had a pH(water) range of between 4.9 and 7.5, with four acidic (pH 4.9 – 5.5), four ‘neutral’ (5.8 – 6.1) and four alkaline (6.5 – 7.5) soils. Bacteria were recovered from nodules of Trifolium repens (white clover) and T. subterraneum (subterranean clover) grown in the soils. The strains were cultured and screened against a range of pH-amended media to demonstrate whether they were adapted to pH levels similar to their native soils. The strains which showed high relative growth at a given pH (~20% of those isolated) were selected for metabolic and taxonomic profiling. The Omnilog (Biolog Inc., Hayward, CA) phenotype array was used to perform assays on carbon (C) utilisation for selected strains. DNA was extracted from the strains which had differing C utilisation profiles and PCR products for both forward and reverse primers were sequenced for the following genes: 16S rRNA, recA, nodC, nodD and nifH (symbiotic).

Keywords: bacterial diversity, clover, metabolic and taxonomic profiling, pH adaptation, rhizobia

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218 Investigating Homicide Offender Typologies Based on Their Clinical Histories and Crime Scene Behaviour Patterns

Authors: Valeria Abreu Minero, Edward Barker, Hannah Dickson, Francois Husson, Sandra Flynn, Jennifer Shaw

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Purpose – The purpose of this paper is to identify offender typologies based on aspects of the offenders’ psychopathology and their associations with crime scene behaviours using data derived from the National Confidential Enquiry into Suicide and Safety in Mental Health concerning homicides in England and Wales committed by offenders in contact with mental health services in the year preceding the offence (n=759). Design/methodology/approach – The authors used multiple correspondence analysis to investigate the interrelationships between the variables and hierarchical agglomerative clustering to identify offender typologies. Variables describing: the offender’s mental health history; the offenders’ mental state at the time of offence; characteristics useful for police investigations; and patterns of crime scene behaviours were included. Findings – Results showed differences in the offender’s histories in relation to their crime scene behaviours. Further, analyses revealed three homicide typologies: externalising, psychosis and depression. Analyses revealed three homicide typologies: externalising, psychotic and depressive. Practical implications – These typologies may assist the police during homicide investigations by: furthering their understanding of the crime or likely suspect; offering insights into crime patterns; provide advice as to what an offender’s offence behaviour might signify about his/her mental health background; findings suggest information concerning offender psychopathology may be useful for offender profiling purposes in cases of homicide offenders with schizophrenia, depression and comorbid diagnosis of personality disorder and alcohol/drug dependence. Originality/value – Empirical studies with an emphasis on offender profiling have almost exclusively focussed on the inference of offender demographic characteristics. This study provides a first step in the exploration of offender psychopathology and its integration to the multivariate analysis of offence information for the purposes of investigative profiling of homicide by identifying the dominant patterns of mental illness within homicidal behaviour.

Keywords: offender profiling, mental illness, psychopathology, multivariate analysis, homicide, crime scene analysis, crime scene behviours, investigative advice

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217 Effect of Laser Input Energy on the Laser Joining of Polyethylene Terephthalate to Titanium

Authors: Y. J. Chen, T. M. Yue, Z. N. Guo

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This paper reports the effects of laser energy on the characteristics of bubbles generated in the weld zone and the formation of new chemical bonds at the Polyethylene Terephthalate (PET)/Ti joint interface in laser joining of PET to Ti. The samples were produced by using different laser energies ranging from 1.5 J – 6 J in steps of 1.5 J, while all other joining parameters remained unchanged. The types of chemical bonding at the joint interface were analysed by the x-ray photoelectron spectroscopy (XPS) depth-profiling method. The results show that the characteristics of the bubbles and the thickness of the chemically bonded interface, which contains the laser generated bonds of Ti–C and Ti–O, increase markedly with increasing laser energy input. The tensile failure load of the joint depends on the combined effect of the amount and distribution of the bubbles formed and the chemical bonding intensity of the joint interface.

Keywords: laser direct joining, Ti/PET interface, laser energy, XPS depth profiling, chemical bond, tensile failure load

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216 Chemical Profiling of Farsetia Aegyptia Turra and Farsetia Longisiliqua Decne. and Their Chemosystematic Significance

Authors: Mona M. Marzouk, Ahmed Elkhateeb, Mona Elshabrawy, Mai M. Farid, Salwa A. Kawashty, EL-Sayed S. Abdel-Hameed, Sameh R. Hussein

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The genus Farsetia Turra belongs to the family Brassicaceae and has approximately 30 accepted species distributed worldwide. Amongst them, Farsetia aegyptia Turra and Farsetia longisiliqua Decne. are two common species characteristic to the Egyptian flora. The present study considers the first characterization of the chemical constituents of F. longisiliqua, aiming to compare with those identified from the medicinal species (F. aegyptia). Additionally, the chemosystematic relationships between the two studied species were evaluated and highlight the medicinal importance for F. longisiliqua. The chemical profiling of their aqueous methanol extracts were carried out using the LC-ESI-MS technique and afforded 54 compounds belonging to different chemical groups. Flavonoids are the major constituents and are represented by 32 compounds (two C-glycosyl flavone, four flavones, and 26 flavonols). Their structural variations and common constituents confirmed the chemosystematic significance of the two species. Moreover, the flavonoid profiles showed major common constituents between the two investigated species, which predicted the medicinal importance of F. longisiliqua.

Keywords: brassicaceae, chemosystematics, farsetia, flavonoids, glucosinolates, LC-ESI-MS

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215 NMR-Based Metabolomics Reveals Dietary Effects in Liver Extracts of Arctic Charr (Salvelinus alpinus) and Tilapia (Oreochromis mossambicus) Fed Different Levels of Starch

Authors: Rani Abro, Ali Ata Moazzami, Jan Erik Lindberg, Torbjörn Lundh

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The effect of dietary starch level on liver metabolism in Arctic charr (Salvelinus alpinus) and tilapia (Oreochromis mossambicus) was studied using 1H-NMR based metabolomics. Fingerlings were fed iso-nitrogenous diets containing 0, 10 and 20 % starch for two months before liver samples were collected for metabolite analysis. Metabolite profiling was performed using 600 MHz NMR Chenomx software. In total, 48 metabolites were profiled in liver extracts from both fish species. Following the profiling, principal component analysis (PCA) and orthogonal partial least square discriminant analysis (OPLC-DA) were performed. These revealed that differences in the concentration of significant metabolites were correlated to the dietary starch level in both species. The most prominent difference in metabolic response to starch feeding between the omnivorous tilapia and the carnivorous Arctic charr was an indication of higher anaerobic metabolism in Arctic charr. The data also indicated that amino acid and pyrimidine metabolism was higher in Artic charr than in tilapia.

Keywords: arctic charr, metabolomics, starch, tilapia

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214 Comparative Transcriptome Profiling of Low Light Tolerant and Sensitive Rice Varieties Induced by Low Light Stress at Active Tillering Stage

Authors: Darshan Panda, Lambodar Behera, M. J. Baig, Sudhanshu Sekhar

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Low light intensity is a significant limitation for grain yield and quality in rice. However, yield is not significantly reduced in low-light tolerant rice varieties. The work, therefore, planned for comparative transcriptome profiling under low light stress to decipher the genes involved and molecular mechanism of low light tolerance in rice. At the active tillering stage, 50% low light exposure for one day, three days, and five days were given to Swarnaprabha (low light tolerant) and IR8 (low light sensitive) rice varieties. Illumina (HiSeq) platform was used for transcriptome sequencing. A total of 6,652 and 12,042 genes were differentially expressed due to low light intensity in Swarnaprabha and IR8, respectively, as compared to control. CAB, LRP, SBPase, MT15, TF PCL1, and Photosystem I & II complex related gene expressions were mostly increased in Swarnaprabha upon the longer duration of low light exposure, which was not found in IR8 as compared to control. Their expressions were validated by qRT-PCR. The overall study suggested that the maintenance of grain yield in the tolerant variety under low light might be the result of accelerated expression of the genes, which enable the plant to keep the photosynthetic processes moving at the same pace even under low light.

Keywords: rice, low light, photosynthesis, yield

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213 Impact of Chimerism on Y-STR DNA Determination: Sex Mismatch Analysis

Authors: Anupuma Raina, Ajay P. Balayan, Prateek Pandya, Pankaj Shrivastava, Uma Kanga, Tulika Seth

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DNA fingerprinting analysis aids in personal identification for forensic purposes and has always been a driving motivation for law enforcement agencies in almost all countries since its inception. The introduction of DNA markers (Y-STR) has allowed for greater precision and higher discriminatory power in forensic testing. A criminal/ person committing crime after bone marrow transplantation is a rare situation but not an impossible one. Keeping such a situation in mind, a study was carried out to find out the best biological sample to be used for personal identification, especially in forensic situation. We choose a female patient (recipient) and a male donor. The pre transplant sample (blood) and post transplant samples (blood, buccal swab, hair roots) were collected from the recipient (patient). The same were compared with the blood sample of the donor using DNA FP technique. Post transplant samples were collected at different interval of time (15, 30, 60, and 90 days). The study was carried out using Y-STR kit at 23 loci. The results determined discusses the phenomenon of chimerism and its impact on Y-STR. Hair sample was found the most suitable sample which had no donor DNA profiling up to 90 days.

Keywords: bone marrow transplantation, chimerism, DNA profiling, Y-STR

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212 Lc-Ms N-Alkylamide Profiling of an Ethanolic Anacyclus pyrethrum Root Extract

Authors: Vikas Sharma, V. K. Dixit

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The roots of Anacyclus pyrethrum DC (AP) (Asteraceae) are frequently used in traditional medicine as Vajikarana Rasayana. An ethanolic extract of root of Anacyclus pyrethrum demonstrated its potential to enhance the sexual behaviour of male rats, with a dose dependent effect on sperm count and androgens concentration. Phytochemical analysis of ethanolic extract of Anacyclus pyrethrum revealed that it is rich in N-alkylamide. This study therefore sought to assess characterization of ethanolic extract of Anacyclus pyrethrum root. Root extract was performed using a gradient reversed phase high performance liquid chromatography/UV/electrospray ionization ion trap mass spectrometry (HPLC/ESI-MS) method on an embedded polar column. MS1 and MS2 fragmentation data were used for identification purposes, while UV was used for quantification. Thirteen N-alkylamides (five N-isobutylamides, three N-methyl isobutylamides, four tyramides, and one 2-phenylethylamide) were detected. Five of them identified as undeca-2E,4E-diene-8,10-diynoic acid N-methyl isobutylamide, tetradeca-2E,4E-diene-8,10-diynoic acid tyramide, deca-2E,4E-dienoic acid N-methyl isobutylamide, tetradeca-2E,4E,XE/Z-trienoic acid tyramide and tetradeca-2E,4E,8Z,10Z-tetraenoic isobutylamide are novel compounds, which have never been identified in Anacyclus pyrethrum.

Keywords: Anacyclus pyrethrum (Asteraceae), LC-MS plant profiling, N-alkylamides, pellitorine, anacycline

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211 Molecular Profiling of an Oleaginous Trebouxiophycean Alga Parachlorella kessleri Subjected to Nutrient Deprivation

Authors: Pannaga Pavan Jutur

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Parachlorella kessleri, a marine unicellular green alga belonging to class Trebouxiophyceae, accumulates large amounts of oil, i.e., lipids under nutrient-deprived (-N, -P, and -S) conditions. Understanding their metabolic imprints is important for elucidating the physiological mechanisms of lipid accumulations in this microalga subjected to nutrient deprivation. Metabolic and lipidomic profiles were obtained respectively using gas chromatography-mass spectrometry (GC-MS) of P. kessleri under nutrient starvation (-N, -P and -S) conditions. Relative quantities of more than 100 metabolites were systematically compared in all these three starvation conditions. Our results demonstrate that in lipid metabolism, the quantities of neutral lipids increased significantly followed by the decrease in other metabolites involved in photosynthesis, nitrogen assimilation, etc. In conclusion, the metabolomics and lipidomic profiles have identified a few common metabolites such as citric acid, valine, and trehalose to play a significant role in the overproduction of oil by this microalga subjected to nutrient deprivation. Understanding the entire system through untargeted metabolome profiling will lead to identifying relevant metabolites involved in the biosynthesis and degradation of precursor molecules that may have the potential for biofuel production, aiming towards the vision of tomorrow’s bioenergy needs.

Keywords: algae, biofuels, nutrient stress, omics

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210 HPTLC Fingerprint Profiling of Protorhus longifolia Methanolic Leaf Extract and Qualitative Analysis of Common Biomarkers

Authors: P. S. Seboletswe, Z. Mkhize, L. M. Katata-Seru

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Protorhus longifolia is known as a medicinal plant that has been used traditionally to treat various ailments such as hemiplegic paralysis, blood clotting related diseases, diarrhoea, heartburn, etc. The study reports a High-Performance Thin Layer Chromatography (HPTLC) fingerprint profile of Protorhus longifolia methanolic extract and its qualitative analysis of gallic acid, rutin, and quercetin. HPTLC analysis was achieved using CAMAG HPTLC system equipped with CAMAG automatic TLC sampler 4, CAMAG Automatic Developing Chamber 2 (ADC2), CAMAG visualizer 2, CAMAG Thin Layer Chromatography (TLC) scanner and visionCATS CAMAG HPTLC software. Mobile phase comprising toluene, ethyl acetate, formic acid (21:15:3) was used for qualitative analysis of gallic acid and revealed eight peaks while the mobile phase containing ethyl acetate, water, glacial acetic acid, formic acid (100:26:11:11) for qualitative analysis of rutin and quercetin revealed six peaks. HPTLC sillica gel 60 F254 glass plates (10 × 10) were used as the stationary phase. Gallic acid was detected at the Rf = 0.35; while rutin and quercetin were not evident in the extract. Further studies will be performed to quantify gallic acid in Protorhus longifolia leaves and also identify other biomarkers.

Keywords: biomarkers, fingerprint profiling, gallic acid, HPTLC, Protorhus longifolia

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209 Oncogenic Functions of Long Non-Coding RNA XIST in Human Nasopharyngeal Carcinoma by Targeting MiR-34a-5p

Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li

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Long non-coding RNA (lncRNA) X inactivate-specific transcript (XIST) has been verified as an oncogenic gene in several human malignant tumors, and its dysregulation was closed associated with tumor initiation, development and progression. Nevertheless, whether the aberrant expression of XIST in human nasopharyngeal carcinoma (NPC) is corrected with malignancy, metastasis or prognosis has not been elaborated. Here, we discovered that XIST was up-regulated in NPC tissues and higher expression of XIST contributed to a markedly poorer survival time. In addition, multivariate analysis demonstrated XIST was an independent risk factor for prognosis. XIST over-expression enhanced, while XIST silencing hampered the cell growth in NPC. Additionally, mechanistic analysis revealed that XIST up-regulated the expression of miR-34a-5p targeted gene E2F3 through acting as a competitive ‘sponge’ of miR-34a-5p. Taking all into account, we concluded that XIST functioned as an oncogene in NPC through up-regulating E2F3 in part through ‘spongeing’ miR-34a-5p.

Keywords: X inactivate-specific transcript; hsa-miRNA-34a-5p, miR-34a-5p; E2F3, nasopharyngeal carcinoma, tumorigenesis

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208 Salicylic Acid Signalling in Relation to Root Colonization in Rice

Authors: Seema Garcha, Sheetal Chopra, Navraj Sarao

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Plant hormones play a role in internal colonization by beneficial microbes and also systemic acquired resistance. They define qualitative and quantitative nature of root microbiome and also influence dynamics of root rhizospheric soil. The present study is an attempt to relate salicylic acid (signal molecule) content and qualitative nature of root endophytes at various stages in the growth of rice varieties of commercial value- Parmal 121 and Basmati 1121. Root seedlings of these varieties were raised using tissue culture techniques and then they were transplanted in the fields. Cultivation was done using conventional methods in agriculture. Field soil contained 0.39% N, 75.12 Kg/hectare of phosphorus and 163.0 Kg/hectare of potassium. Microfloral profiling of the root tissue was done using the selective microbiological medium. The salicylic acid content was estimated using HPLC-Agilent 1100 HPLC Series. Salicylic acid level of Basmati 1121 remained relatively low at the time of transplant and 90 days after transplant. It increased marginally at 60 days. A similar trend was observed with Parmal 121 as well. However, Parmal variety recorded 0.935 ug/g of salicylic acid at 60 days after transplant. Salicylic acid content decreased after 90 days as both the rice varieties remained disease free. The endophytic root microflora was established by 60 days after transplant in both the varieties after which their population became constant. Rhizobium spp dominated over Azotobacter spp. Genetic profiling of endophytes for nitrogen-fixing ability is underway.

Keywords: plant-microbe interaction, rice, root microbiome, salicylic acid

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207 Exploring the Strategy to Identify Seed-Specific Acyl-Hydrolases from Arabidopsis thaliana by Activity-Based Protein Profiling

Authors: M. Latha, Achintya K. Dolui, P. Vijayaraj

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Vegetable oils mainly triacylglycerol (TAG) are an essential nutrient in the human diet as well as one of the major global commodity. There is a pressing need to enhance the yield of oil production to meet the world’s growing demand. Oil content is controlled by the balance between synthesis and breakdown in the cells. Several studies have established to increase the oil content by the overexpression of oil biosynthetic enzymes. Interestingly the significant oil accumulation was observed with impaired TAG hydrolysis. Unfortunately, the structural, as well as the biochemical properties of the lipase enzymes, is widely unknown, and so far, no candidate gene was identified in seeds except sugar-dependent1 (SDP1). Evidence has shown that SDP1directly responsible for initiation of oil breakdown in the seeds during germination. The present study is the identification of seed-specific acyl-hydrolases by activity based proteome profiling (ABPP) using Arabidopsis thaliana as a model system. The ABPP reveals that around 8 to 10 proteins having the serine hydrolase domain and are expressed during germination of Arabidopsis seed. The N-term sequencing, as well as LC-MS/MS analysis, was performed for the differentially expressed protein during germination. The coding region of the identified proteins was cloned, and lipases activity was assessed with purified recombinant protein. The enzyme assay was performed against various lipid substrates, and we have observed the acylhydrolase activity towards lysophosphatidylcholine and monoacylglycerol. Further, the functional characteristic of the identified protein will reveal the physiological significance the enzyme in oil accumulation.

Keywords: lipase, lipids, vegetable oil, triacylglycerol

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206 The Metabolite Profiling of Fulvestrant-3 Boronic Acid under Biological Oxidation

Authors: Changde Zhang, Qiang Zhang, Shilong Zheng, Jiawang Liu, Shanchun Guo, Qiu Zhong, Guangdi Wang

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Fulvestrant was approved by FDA to treat breast cancer as a selective estrogen receptor downregulator (SERD) with intramuscular injection administration. ZB716, a fulvestarnt-3 boronic acid, is an SERD with comparable anticancer effect to fulvestrant, but could produce good pharmacokinetic properties under oral administration with mice or rat models. To understand why ZB716 produced much better oral bioavailability, it was proposed that the boronic acid blocked the phase II direct biotransformation with the hydroxyl group on the 3 position of the aromatic ring on fulvestrant. In this study, ZB716 or fulvestrant was incubated with human liver microsome and oxidation cofactor NADPH in vitro. Their metabolites after oxidation were profiled with the Q-Exactive, a high-resolution mass spectrometer. The result showed that ZB716 blocked the forming of hydroxyl groups on its benzene ring except for the oxidation of C-B bond forming fulvestrant in its metabolites, and the concentration of fulvestrant with one more hydroxyl group found in the metabolites from incubation with fulvestrant was about 34 fold high as that formed from incubation with ZB716. Compared to fulvestrant, ZB716 is expected to be much difficult to be further bio-transformed into more hydrophilic compounds, to be difficult excreted out of blood system, and to have longer residence time in blood, which can lead to higher oral bioavailability. This study provided evidence to explain the high bioavailability of ZB716 after oral administration from the perspective of its difficulty of oxidation, a phase I biotransformation, on positions on its aromatic ring.

Keywords: biotransformation, fulvestrant, metabolite profiling, ZB716

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205 Design of an Automated Deep Learning Recurrent Neural Networks System Integrated with IoT for Anomaly Detection in Residential Electric Vehicle Charging in Smart Cities

Authors: Wanchalerm Patanacharoenwong, Panaya Sudta, Prachya Bumrungkun

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The paper focuses on the development of a system that combines Internet of Things (IoT) technologies and deep learning algorithms for anomaly detection in residential Electric Vehicle (EV) charging in smart cities. With the increasing number of EVs, ensuring efficient and reliable charging systems has become crucial. The aim of this research is to develop an integrated IoT and deep learning system for detecting anomalies in residential EV charging and enhancing EV load profiling and event detection in smart cities. This approach utilizes IoT devices equipped with infrared cameras to collect thermal images and household EV charging profiles from the database of Thailand utility, subsequently transmitting this data to a cloud database for comprehensive analysis. The methodology includes the use of advanced deep learning techniques such as Recurrent Neural Networks (RNN) and Long Short-Term Memory (LSTM) algorithms. IoT devices equipped with infrared cameras are used to collect thermal images and EV charging profiles. The data is transmitted to a cloud database for comprehensive analysis. The researchers also utilize feature-based Gaussian mixture models for EV load profiling and event detection. Moreover, the research findings demonstrate the effectiveness of the developed system in detecting anomalies and critical profiles in EV charging behavior. The system provides timely alarms to users regarding potential issues and categorizes the severity of detected problems based on a health index for each charging device. The system also outperforms existing models in event detection accuracy. This research contributes to the field by showcasing the potential of integrating IoT and deep learning techniques in managing residential EV charging in smart cities. The system ensures operational safety and efficiency while also promoting sustainable energy management. The data is collected using IoT devices equipped with infrared cameras and is stored in a cloud database for analysis. The collected data is then analyzed using RNN, LSTM, and feature-based Gaussian mixture models. The approach includes both EV load profiling and event detection, utilizing a feature-based Gaussian mixture model. This comprehensive method aids in identifying unique power consumption patterns among EV owners and outperforms existing models in event detection accuracy. In summary, the research concludes that integrating IoT and deep learning techniques can effectively detect anomalies in residential EV charging and enhance EV load profiling and event detection accuracy. The developed system ensures operational safety and efficiency, contributing to sustainable energy management in smart cities.

Keywords: cloud computing framework, recurrent neural networks, long short-term memory, Iot, EV charging, smart grids

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204 Blood Chemo-Profiling in Workers Exposed to Occupational Pyrethroid Pesticides to Identify Associated Diseases

Authors: O. O. Sufyani, M. E. Oraiby, S. A. Qumaiy, A. I. Alaamri, Z. M. Eisa, A. M. Hakami, M. A. Attafi, O. M. Alhassan, W. M. Elsideeg, E. M. Noureldin, Y. A. Hobani, Y. Q. Majrabi, I. A. Khardali, A. B. Maashi, A. A. Al Mane, A. H. Hakami, I. M. Alkhyat, A. A. Sahly, I. M. Attafi

Abstract:

According to the Food and Agriculture Organization (FAO) Pesticides Use Database, pesticide use in agriculture in Saudi Arabia has more than doubled from 4539 tons in 2009 to 10496 tons in 2019. Among pesticides, pyrethroids is commonly used in Saudi Arabia. Pesticides may increase susceptibility to a variety of diseases, particularly among pesticide workers, due to their extensive use, indiscriminate use, and long-term exposure. Therefore, analyzing blood chemo-profiles and evaluating the detected substances as biomarkers for pyrethroid pesticide exposure may assist to identify and predicting adverse effects of exposure, which may be used for both preventative and risk assessment purposes. The purpose of this study was to (a) analyze chemo-profiling by Gas Chromatography-Mass Spectrometry (GC-MS) analysis, (b) identify the most commonly detected chemicals in a time-exposure-dependent manner using a Venn diagram, and (c) identify their associated disease among pesticide workers using analyzer tools on the Comparative Toxicogenomics Database (CTD) website, (250 healthy male volunteers (20-60 years old) who deal with pesticides in the Jazan region of Saudi Arabia (exposure intervals: 1-2, 4-6, 6-8, more than 8 years) were included in the study. A questionnaire was used to collect demographic information, the duration of pesticide exposure, and the existence of chronic conditions. Blood samples were collected for biochemistry analysis and extracted by solid-phase extraction for gas chromatography-mass spectrometry (GC-MS) analysis. Biochemistry analysis reveals no significant changes in response to the exposure period; however, an inverse association between the albumin level and the exposure interval was observed. The blood chemo-profiling was differentially expressed in an exposure time-dependent manner. This analysis identified the common chemical set associated with each group and their associated significant occupational diseases. While some of these chemicals are associated with a variety of diseases, the distinguishing feature of these chemically associated disorders is their applicability for prevention measures. The most interesting finding was the identification of several chemicals; erucic acid, pelargonic acid, alpha-linolenic acid, dibutyl phthalate, diisobutyl phthalate, dodecanol, myristic Acid, pyrene, and 8,11,14-eicosatrienoic acid, associated with pneumoconiosis, asbestosis, asthma, silicosis and berylliosis. Chemical-disease association study also found that cancer, digestive system disease, nervous system disease, and metabolic disease were the most often recognized disease categories in the common chemical set. The hierarchical clustering approach was used to compare the expression patterns and exposure intervals of the chemicals found commonly. More study is needed to validate these chemicals as early markers of pyrethroid insecticide-related occupational disease, which might assist evaluate and reducing risk. The current study contributes valuable data and recommendations to public health.

Keywords: occupational, toxicology, chemo-profiling, pesticide, pyrethroid, GC-MS

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203 Qualitative Profiling Model and Competencies Evaluation to Fighting Unemployment

Authors: Francesca Carta, Giovanna Linfante, Laura Agneni, Debora Radicchia, Camilla Micheletta, Angelo Del Cimmuto

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Overtaking competence mismatches and fostering career pathways congruent with the individual skills profile would significantly contribute to fighting unemployment. The aim of this paper is to examine the usefulness and efficiency of qualitative tools in supporting and improving the quality of caseworkers’ activities during the jobseekers’ profile analysis and career guidance process. The selected target groups are long-term and middle term unemployed, job seekers, young people at the end of the vocational training pathway and unemployed woman with social disadvantages. The experimentation is conducted in Italy at public employment services in 2017. In the framework of Italian labour market reform, the experimentation represents the first step to develop a customized qualitative model profiling; the final general object is to improve the public employment services quality. The experimentation tests the transferability of an OECD self-assessment competences tool in the Italian public employment services. On one hand, the first analysis results will indicate the user’s perception concerning the tool’s application and their different competence levels (literacy, numeracy, problem solving, career interest, subjective well-being and health, behavioural competencies) with reference to the specific target. On the other hand, the experimentation outcomes will show caseworkers understanding regarding the tool’s usability and efficiency for career guidance and reskilling and upskilling programs.

Keywords: career guidance, evaluation competences, reskilling pathway, unemployment

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202 Author Profiling: Prediction of Learners’ Gender on a MOOC Platform Based on Learners’ Comments

Authors: Tahani Aljohani, Jialin Yu, Alexandra. I. Cristea

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The more an educational system knows about a learner, the more personalised interaction it can provide, which leads to better learning. However, asking a learner directly is potentially disruptive, and often ignored by learners. Especially in the booming realm of MOOC Massive Online Learning platforms, only a very low percentage of users disclose demographic information about themselves. Thus, in this paper, we aim to predict learners’ demographic characteristics, by proposing an approach using linguistically motivated Deep Learning Architectures for Learner Profiling, particularly targeting gender prediction on a FutureLearn MOOC platform. Additionally, we tackle here the difficult problem of predicting the gender of learners based on their comments only – which are often available across MOOCs. The most common current approaches to text classification use the Long Short-Term Memory (LSTM) model, considering sentences as sequences. However, human language also has structures. In this research, rather than considering sentences as plain sequences, we hypothesise that higher semantic - and syntactic level sentence processing based on linguistics will render a richer representation. We thus evaluate, the traditional LSTM versus other bleeding edge models, which take into account syntactic structure, such as tree-structured LSTM, Stack-augmented Parser-Interpreter Neural Network (SPINN) and the Structure-Aware Tag Augmented model (SATA). Additionally, we explore using different word-level encoding functions. We have implemented these methods on Our MOOC dataset, which is the most performant one comparing with a public dataset on sentiment analysis that is further used as a cross-examining for the models' results.

Keywords: deep learning, data mining, gender predication, MOOCs

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201 The Reliability and Shape of the Force-Power-Velocity Relationship of Strength-Trained Males Using an Instrumented Leg Press Machine

Authors: Mark Ashton Newman, Richard Blagrove, Jonathan Folland

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The force-velocity profile of an individual has been shown to influence success in ballistic movements, independent of the individuals' maximal power output; therefore, effective and accurate evaluation of an individual’s F-V characteristics and not solely maximal power output is important. The relatively narrow range of loads typically utilised during force-velocity profiling protocols due to the difficulty in obtaining force data at high velocities may bring into question the accuracy of the F-V slope along with predictions pertaining to the maximum force that the system can produce at a velocity of null (F₀) and the theoretical maximum velocity against no load (V₀). As such, the reliability of the slope of the force-velocity profile, as well as V₀, has been shown to be relatively poor in comparison to F₀ and maximal power, and it has been recommended to assess velocity at loads closer to both F₀ and V₀. The aim of the present study was to assess the relative and absolute reliability of an instrumented novel leg press machine which enables the assessment of force and velocity data at loads equivalent to ≤ 10% of one repetition maximum (1RM) through to 1RM during a ballistic leg press movement. The reliability of maximal and mean force, velocity, and power, as well as the respective force-velocity and power-velocity relationships and the linearity of the force-velocity relationship, were evaluated. Sixteen male strength-trained individuals (23.6 ± 4.1 years; 177.1 ± 7.0 cm; 80.0 ± 10.8 kg) attended four sessions; during the initial visit, participants were familiarised with the leg press, modified to include a mounted force plate (Type SP3949, Force Logic, Berkshire, UK) and a Micro-Epsilon WDS-2500-P96 linear positional transducer (LPT) (Micro-Epsilon, Merseyside, UK). Peak isometric force (IsoMax) and a dynamic 1RM, both from a starting position of 81% leg length, were recorded for the dominant leg. Visits two to four saw the participants carry out the leg press movement at loads equivalent to ≤ 10%, 30%, 50%, 70%, and 90% 1RM. IsoMax was recorded during each testing visit prior to dynamic F-V profiling repetitions. The novel leg press machine used in the present study appears to be a reliable tool for measuring F and V-related variables across a range of loads, including velocities closer to V₀ when compared to some of the findings within the published literature. Both linear and polynomial models demonstrated good to excellent levels of reliability for SFV and F₀ respectively, with reliability for V₀ being good using a linear model but poor using a 2nd order polynomial model. As such, a polynomial regression model may be most appropriate when using a similar unilateral leg press setup to predict maximal force production capabilities due to only a 5% difference between F₀ and obtained IsoMax values with a linear model being best suited to predict V₀.

Keywords: force-velocity, leg-press, power-velocity, profiling, reliability

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200 Role of Long Noncoding RNA HULC on Colorectal Carcinoma Progression through Epigenetically Repressing NKD2 Expression

Authors: Shu-Jun Li, Cheng-Cao Sun, De-Jia Li

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Recently, long noncoding RNAs (lncRNAs) have been emerged as crucial regulators of human diseases and prognostic markers in numerous of cancers, including colorectal carcinoma (CRC). Here, we identified an oncogenetic lncRNA HULC, which may promote colorectal tumorigenesis. HULC has been found to be up-regulated and acts as oncogene in gastric cancer and hepatocellular carcinoma, but its expression pattern, biological function and underlying mechanism in CRC is still undetermined. Here, we reported that HULC expression is also over-expressed in CRC, and its increased level is associated with poor prognosis and shorter survival. Knockdown of HULC impaired CRC cells proliferation, migration and invasion, facilitated cell apoptosis in vitro, and inhibited tumorigenicity of CRC cells in vivo. Mechanistically, RNA immunoprecipitation (RIP) and RNA pull-down experiment demonstrated that HULC could simultaneously interact with EZH2 to repress underlying targets NKD2 transcription. In addition, rescue experiments determined that HULC oncogenic function is partly dependent on repressing NKD2. Taken together, our findings expound how HULC over-expression endows an oncogenic function in CRC.

Keywords: long noncoding RNA, HULC, NKD2, colorectal carcinoma, proliferation, apoptosis

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199 Potential of Croatia as an Attractive Tourist Destination for the Russian Market

Authors: Maja Martinovic, Valentina Zarkovic, Hrvoje Maljak

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Europe is one of the most popular tourist destinations in the world, in which tourism occupies a significant place among the most relevant economic activities, and this applies to the Republic of Croatia as well. Based on this study, the authors intended to encourage and support the creation of an effective tourism policy in Croatia that would be based on the profiling of certain target groups. Another objective was to compare the results obtained from the customer analysis with the market analysis of the tourism industry in Croatia. The objective is to adapt the current tourist offer according to the identified needs and expectations of a particular tourist group in order to increase the attractiveness of Croatia as a tourist destination and motivate greater attendance of the targeted tourist groups. The current research was oriented towards the Russian market as the target group. Therefore, the authors wanted to encourage a discussion on how to attract more Russian guests. Consequently, the intention of the research was a detailed analysis of Russian tourists, in order to gain a better understanding of their travelling motives and tendencies. Furthermore, attention was paid to the expectations of Russian customers and to compare them with the Croatian tourist offer, and to determine whether there is a possibility for an overlap. The method used to obtain the information required was a survey conducted among Russian citizens about their travelling habits. The research was carried out on the basis of 166 participants of different age, gender, profession and income group. The sampling and distribution of the survey took place between May and July 2016. The results provided from the research indicate that Croatian tourism has certain unrealized potential considering the popularization of Croatia as a tourist destination, and there is a capacity for increasing the revenues within the group of Russian tourists. Such a conclusion is based on the fact that the Croatian tourist offer and the preferences of the Russian guests are compatible, i.e. they overlap in many aspects. The results demonstrate that beautiful nature, cultural and historical heritage as well as the sun and sea, play a leading role in attracting more Russian tourists. It is precisely these elements that form the three pillars of the Croatian tourist offer. On the other hand, the profiling revealed that the most desirable destinations for the Russian guests are Italy and Spain, both of which provide the same main tourist attractions as Croatia. Therefore, the focus of the strategic ideas given in the paper shifted to other tourism segments, such as type of accommodation, sales channels, travel motives, additional offer and seasonality etc., in order to gain advantage in the Russian market, the Mediterranean region and tourism in general. The purpose of the research is to serve as a foundation for analysing the attractiveness of the other tourist destinations in the Russian market, as well as to be a general basis for a more detailed profiling of the various specific target groups of the Russian and other tourist groups.

Keywords: Croatia, Russian market, target groups, tourism, tourist destination

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198 Nanoprofiling of GaAs Surface in a Combined Low-Temperature Plasma for Microwave Devices

Authors: Victor S. Klimin, Alexey A. Rezvan, Maxim S. Solodovnik, Oleg A. Ageev

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In this paper, the problems of existing methods of profiling and surface modification of nanoscale arsenide-gallium structures are analyzed. The use of a combination of methods of local anodic oxidation and plasma chemical etching to solve this problem is considered. The main features that make this technology one of the promising areas of modification and profiling of near-surface layers of solids are demonstrated. In this paper, we studied the effect of formation stress and etching time on the geometrical parameters of the etched layer and the roughness of the etched surface. Experimental dependences of the thickness of the etched layer on the time and stress of formation were obtained. The surface analysis was carried out using atomic force microscopy methods, the corresponding profilograms were constructed from the obtained images, and the roughness of the etched surface was studied accordingly. It was shown that at high formation voltage, the depth of the etched surface increased, this is due to an increase in the number of active particles (oxygen ions and hydroxyl groups) formed as a result of the decomposition of water molecules in an electric field, during the formation of oxide nanostructures on the surface of gallium arsenide. Oxide layers were used as negative masks for subsequent plasma chemical etching by the STE ICPe68 unit. BCl₃ was chosen as the chlorine-containing gas, which differs from analogs in some parameters for the effect of etching of nanostructures based on gallium arsenide in the low-temperature plasma. The gas mixture of reaction chamber consisted of a buffer gas NAr = 100 cm³/min and a chlorine-containing gas NBCl₃ = 15 cm³/min at a pressure P = 2 Pa. The influence of these methods modes, which are formation voltage and etching time, on the roughness and geometric parameters, and corresponding dependences are demonstrated. Probe nanotechnology was used for surface analysis.

Keywords: nanostructures, GaAs, plasma chemical etching, modification structures

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197 Identification of Bioactive Metabolites from Ficus carica and Their Neuroprotective Effects of Alzheimer's Disease

Authors: Hanan Khojah, RuAngelie Edrada-Ebel

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Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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196 Gene Expression Profiling of Iron-Related Genes of Pasteurella multocida Serotype A Strain PMTB2.1

Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar

Abstract:

Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.

Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes

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