Search results for: whole genome sequence

Authors: Graham Hancox, Saydia Razak, Sue Hignett, Jo Barnes, Jyri Silmari, Florian Kading

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In the event of a Chemical, Biological, Radiological and Nuclear (CBRN) incident rapidly gaining, situational awareness is of paramount importance and advanced technologies have an important role to play in improving detection, identification, monitoring (DIM) and patient tracking. Understanding how these advanced technologies can fit into current response systems is essential to ensure they are optimally designed, usable and meet end-users’ needs. For this reason, Human Factors (Ergonomics) methods have been used within an EU Horizon 2020 project (TOXI-Triage) to firstly describe (map) the hierarchical structure in a CBRN response with adapted Accident Map (AcciMap) methodology. Secondly, Hierarchical Task Analysis (HTA) has been used to describe and review the sequence of steps (sub-tasks) in a CBRN scenario response as a task system. HTA methodology was then used to map one advanced technology, ‘Tag and Trace’, which tags an element (people, sample and equipment) with a Near Field Communication (NFC) chip in the Hot Zone to allow tracing of (monitoring), for example casualty progress through the response. This HTA mapping of the Tag and Trace system showed how the provider envisaged the technology being used, allowing for review and fit with the current CBRN response systems. These methodologies have been found to be very effective in promoting and supporting a dialogue between end-users and technology providers. The Human Factors methods have given clear diagrammatic (visual) representations of how providers see their technology being used and how end users would actually use it in the field; allowing for a more user centered approach to the design process. For CBRN events usability is critical as sub-optimum design of technology could add to a responders’ workload in what is already a chaotic, ambiguous and safety critical environment.

Keywords: AcciMap, CBRN, ergonomics, hierarchical task analysis, human factors

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Authors: Sina Mahdavi, Mahdi Asghari Ozma

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Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human Varicella-zoster virus (VZV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on VZV retrovirus infection in MS disease progression. For this study, the keywords "Multiple sclerosis", " Human Varicella-zoster virus ", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched and 14 articles were chosen, studied, and analyzed. Analysis of the amino acid sequences of HNRNPA1 with VZV proteins has shown a 62% amino acid sequence similarity between VZV gE and the PrLD/M9 epitope region (TNPO1 binding domain) of mutant HNRNPA1. A heterogeneous nuclear ribonucleoprotein (hnRNP), which is produced by HNRNPA1, is involved in the processing and transfer of mRNA and pre-mRNA. Mutant HNRNPA1 mimics gE of VZV as an antigen that leads to autoantibody production. Mutant HnRNPA1 translocates to the cytoplasm, after aggregation is presented by MHC class I, followed by CD8 + cells. Of these, antibodies and immune cells against the gE epitopes of VZV remain due to the memory immune response, causing neurodegeneration and the development of MS in genetically predisposed individuals. VZV expression during the course of MS is present in genetically predisposed individuals with HNRNPA1 mutation, suggesting a link between VZV and MS, and that this virus may play a role in the development of MS by inducing an inflammatory state. Therefore, measures to modulate VZV expression may be effective in reducing inflammatory processes in demyelinated areas of MS patients in genetically predisposed individuals.

Keywords: multiple sclerosis, varicella-zoster virus, central nervous system, autoimmunity

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Authors: Alisa Kazarina, Guntis Gerhards, Elina Petersone-Gordina, Ilva Pole, Viktorija Igumnova, Janis Kimsis, Valentina Capligina, Renate Ranka

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Human body is inhabited by a vast number of microorganisms, collectively known as the human microbiome, and there is a tremendous interest in evolutionary changes in human microbial ecology, diversity and function. The field of paleomicrobiology, study of ancient human microbiome, is powered by modern techniques of Next Generation Sequencing (NGS), which allows extracting microbial genomic data directly from archaeological sample of interest. One of the major techniques is 16S rRNA gene sequencing, by which certain 16S rRNA gene hypervariable regions are being amplified and sequenced. However, some limitations of this method exist including the taxonomic precision and efficacy of different regions used. The aim of this study was to evaluate the phylogenetic sensitivity of different 16S rRNA gene hypervariable regions for microbiome studies in the archaeological samples. Towards this aim, archaeological bone samples and corresponding soil samples from each burial environment were collected in Medieval cemeteries in Latvia. The Ion 16S™ Metagenomics Kit targeting different 16S rRNA gene hypervariable regions was used for library construction (Ion Torrent technologies). Sequenced data were analysed by using appropriate bioinformatic techniques; alignment and taxonomic representation was done using Mothur program. Sequences of most abundant genus were further aligned to E. coli 16S rRNA gene reference sequence using MEGA7 in order to identify the hypervariable region of the segment of interest. Our results showed that different hypervariable regions had different discriminatory power depending on the groups of microbes, as well as the nature of samples. On the basis of our results, we suggest that wider range of primers used can provide more accurate recapitulation of microbial communities in archaeological samples. Acknowledgements. This work was supported by the ERAF grant Nr. 1.1.1.1/16/A/101.

Keywords: 16S rRNA gene, ancient human microbiome, archaeology, bioinformatics, genomics, microbiome, molecular biology, next-generation sequencing

Procedia PDF Downloads 190

Authors: Amr A. El Hanafy, Muhammad I. Qureshi, Jamal Sabir, Mohamed Mutawakil, Mohamed M. Ahmed, Hassan El Ashmaoui, Hassan Ramadan, Mohamed Abou-Alsoud, Mahmoud Abdel Sadek

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β-Lactoglobulin (β-LG) is the dominant non-casein whey protein found in bovine milk and of most ruminants. The amino acid sequence of β-LG along with its 3-dimensional structure illustrates linkage with the lipocalin superfamily. Preliminary studies in goats indicated that milk yield can be influenced by polymorphism in genes coding for whey proteins. The aim of this study is to identify and evaluate the incidence of functional polymorphisms in the exonic and intronic portions of β-LG gene in native Saudi goat breeds (Ardi, Habsi, and Harri). Blood samples were collected from 300 animals (100 for each breed) and genomic DNA was extracted using QIAamp DNA extraction Kit. A fragment of the β-LG gene from exon 7 to 3’ flanking region was amplified with pairs of specific primers. Subsequent digestion with Sac II restriction endonuclease revealed two alleles (A and B) and three different banding patterns or genotypes i.e. AA, AB and BB. The statistical analysis showed that β-LG AA genotype had higher milk yield than β-LG AB and β-LG BB genotypes. Nucleotide sequencing of the selected β-LG fragments was done and submitted to GenBank NCBI (Accession No. KJ544248, KJ588275, KJ588276, KJ783455, KJ783456 and KJ874959). Two already established SNPs in exon 7 (+4601 and +4603) and one fresh SNP in the 3’ UTR region were detected in the β-LG fragments with designated AA genotype. The polymorphisms in exon 7 did not produce any amino acid change. Phylogenetic analysis on the basis of nucleotide sequences of native Saudi goats indicated evolutional similarity with the GenBank reference sequences of goat, Bubalus bubalis and Bos taurus.

Keywords: β-Lactoglobulin, Saudi goats, PCR-RFLP, functional polymorphism, nucleotide sequencing, phylogenetic analysis

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Authors: Reza Miresmaeili, Asghar Heydari Astaraee, Fereshteh Dolati

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In the present study, an analytical model based on dislocation density model was developed to simulate grain refinement in surface mechanical attrition treatment (SMAT). The correlation between SMAT time and development in plastic strain on one hand, and dislocation density evolution, on the other hand, was established to simulate the grain refinement in SMAT. A dislocation density-based constitutive material law was implemented using VUHARD subroutine. A random sequence of shots is taken into consideration for multiple impacts model using Python programming language by utilizing a random function. The simulation technique was to model each impact in a separate run and then transferring the results of each run as initial conditions for the next run (impact). The developed Finite Element (FE) model of multiple impacts describes the coverage evolution in SMAT. Simulations were run to coverage levels as high as 4500%. It is shown that the coverage implemented in the FE model is equal to the experimental coverage. It is depicted that numerical SMAT coverage parameter is adequately conforming to the well-known Avrami model. Comparison between numerical results and experimental measurements for residual stresses and depth of deformation layers confirms the performance of the established FE model for surface engineering evaluations in SMA treatment. X-ray diffraction (XRD) studies of grain refinement, including resultant grain size and dislocation density, were conducted to validate the established model. The full width at half-maximum in XRD profiles can be used to measure the grain size. Numerical results and experimental measurements of grain refinement illustrate good agreement and show the capability of established FE model to predict the gradient microstructure in SMA treatment.

Keywords: dislocation density, grain refinement, severe plastic deformation, simulation, surface mechanical attrition treatment

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Authors: Erika De Finis, Paola Gattinoni, Laura Scesi

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Massive rock avalanches formed some of the largest landslide deposits on Earth and they represent one of the major geohazards in high-relief mountains. This paper interprets a very large sedimentary fan (the Sernio fan, Valtellina, Northern Italy), located 20 Km SW from Val Pola Rock avalanche (1987), as the deposit of a partial collapse of a Deep Seated Gravitational Slope Deformation (DSGSD), afterwards eroded and buried by debris flows. The proposed emplacement sequence has been reconstructed based on geomorphological, structural and mechanical evidences. The Sernio fan is actually considered anomalous with reference to the very high ratio between the fan area (about 4.5km2) and the basin area (about 3km2). The morphology of the fan area is characterised by steep slopes (dip about 20%) and the fan apex is extended for 1.8 km inside the small catchment basin. This sedimentary fan was originated by a landslide that interested a part of a large deep-seated gravitational slope deformation, involving a wide area of about 55 km². The main controlling factor is tectonic and it is related to the proximity to regional fault systems and the consequent occurrence of fault weak rocks (GSI locally lower than 10 with compressive stress lower than 20MPa). Moreover, the fan deposit shows sedimentary evidences of recent debris flow events. The best current explanation of the Sernio fan involves an initial failure of some hundreds of Mm3. The run-out was quite limited because of the morphology of Valtellina’s valley floor, and the deposit filled the main valley forming a landslide dam, as confirmed by the lacustrine deposits detected upstream the fan. Nowadays the debris flow events represent the main hazard in the study area.

Keywords: anomalous sedimentary fans, deep seated gravitational slope deformation, Italy, rock avalanche

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Authors: Catarina Isabel Ramos Pereira, Jorge Machado, Begona Alonso Criado, Maria João Santos

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Introduction: Gait disorders are one of the symptoms that have severe implications on the quality of life in Parkinson's disease (PD). Currently, there is no therapy to reverse or treat this condition. None of the drugs used in conventional medical treatment is entirely efficient, and all have a high incidence of side effects. Acupuncture treatment is believed to improve motor ability, but there is still little scientific evidence in individuals with PD. Aim: The aim of the study is to investigate the acute effect of acupuncture on gait parameters in Parkinson's disease. Methods: This is a randomized and controlled crossover study. The same individual patient was part of both the experimental (real acupuncture) and control group (false acupuncture/sham), and the sequence was randomized. Gait parameters were measured at two different moments, before and after treatment, using four force platforms as well as the collection of 3D markers positions taken by 11 cameras. Images were quantitatively analyzed using Qualisys Track Manager software that let us extract data related to the quality of gait and balance. Seven patients with the diagnosis of Parkinson's disease were included in the study. Results: Statistically significant differences were found in gait speed (p = 0.016), gait cadence (p = 0.006), support base width (p = 0.0001), medio-lateral oscillation (p = 0.017), left-right step length (p = 0.0002), and stride length: right-right (p = 0.0000) and left-left (p = 0.0018), time of left support phase (p = 0.029), right support phase (p = 0.025) and double support phase (p = 0.015), between the initial and final moments for the experimental group. Differences in right-left stride length were found for both groups. Conclusion: Our results show that acupuncture could enhance gait in Parkinson's disease patients. Deep research involving a larger number of volunteers should be accomplished to validate these encouraging findings.

Keywords: acupuncture, traditional Chinese medicine, Parkinson's disease, gait

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Authors: Katharigatta N. Venugopala, Fernando Albericio, Bander E. Al-Dhubiab, T. Govender

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Recent years have witnessed a renaissance in the field of peptides that are obtained from various natural sources such as many bacteria, fungi, plants, seaweeds, vertebrates, invertebrates and have been reported for various pharmacological properties such as anti-TB, anticancer, antimalarial, anti-inflammatory, anti-HIV, antibacterial, antifungal, and antidiabetic, activities. In view of the pharmacological significance of natural peptides, serious research efforts of many scientific groups and pharmaceutical companies have consequently focused on them to explore the possibility of developing their potential analogues as therapeutic agents. Solid phase and solution phase peptide synthesis are the two methodologies currently available for the synthesis of natural or synthetic linear or cyclic depsi-peptides. From a synthetic point of view, there is no doubt that the solid-phase methodology gained added advantages over solution phase methodology in terms of simplicity, purity of the compound and the speed with which peptides can be synthesised. In the present study total synthesis, purification and structural elucidation of analogues of natural anti-TB cyclic depsi-peptides such as depsidomycin, massetolides and viscosin has been attempted by solid phase method using standard Fmoc protocols and finally off resin cyclization in solution phase method. In case of depsidomycin, synthesis of linear peptide on solid phase could not be achieved because of two turn inducing amino acids in the peptide sequence, but total synthesis was achieved by convergent solid phase peptide synthesis followed by cyclization in solution phase method. The title compounds obtained were in good yields and characterized by NMR and HRMS. Anti-TB results revealed that the potential title compound exhibited promising activity at 4 µg/mL against H37Rv and 16 µg/mL against MDR strains of tuberculosis.

Keywords: total synthesis, cyclic depsi-peptides, anti-TB activity, tuberculosis

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Authors: Lixin Tian, Wei Xue

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Many jobs in society go underground, such as mine mining, tunnel construction and subways, which are vital to the development of society. Once accidents occur in these places, the interruption of traditional wired communication is not conducive to the development of rescue work. In order to realize the positioning, early warning and command functions of underground personnel and improve rescue efficiency, it is necessary to develop and design an emergency ground communication system. It is easy to be subjected to narrowband interference when performing conventional underground communication. Spreading communication can be used for this problem. However, general spread spectrum methods such as direct spread communication are inefficient, so it is proposed to use parallel combined spread spectrum (PCSS) communication to improve efficiency. The PCSS communication not only has the anti-interference ability and the good concealment of the traditional spread spectrum system, but also has a relatively high frequency band utilization rate and a strong information transmission capability. So, this technology has been widely used in practice. This paper presents a PCSS communication model-multiple detection parallel combined spread spectrum (MDPCSS) communication system. In this paper, the principle of MDPCSS communication system is described, that is, the sequence at the transmitting end is processed in blocks and cyclically shifted to facilitate multiple detection at the receiving end. The block diagrams of the transmitter and receiver of the MDPCSS communication system are introduced. At the same time, the calculation formula of the system bit error rate (BER) is introduced, and the simulation and analysis of the BER of the system are completed. By comparing with the common parallel PCSS communication, we can draw a conclusion that it is indeed possible to reduce the BER and improve the system performance. Furthermore, the influence of different pseudo-code lengths selected on the system BER is simulated and analyzed, and the conclusion is that the larger the pseudo-code length is, the smaller the system error rate is.

Keywords: cyclic shift, multiple detection, parallel combined spread spectrum, PN code

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Authors: Jernice S. Y. Tan

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Introduction: Reliability within and between testers is vital to ensure the accuracy of any motor assessment instrument. However, reliability checks of the Movement Assessment Battery for Children-2 (MABC-2) age band 3 using multiple testers assigned to different MABC-2 tasks for the same group of participants are uncommon. Multiple testers were not stated as a choice in the MABC-2 manual. Therefore, the purpose of this study was to determine the inter- and intra-tester reliability for using multiple testers to administer the test protocols of MABC-2 age band 3. Methods: Thirty volunteered adolescents (n = 30; 15 males, 15 females; age range: 13 – 16 years) performed the eight tasks in a randomised sequence at three different test stations for the MABC-2 task components (Manual Dexterity, Aiming and Catching, Balance). Ethics approval and parental consent were obtained. The participants were videotaped while performing the test protocols of MABC-2 age band 3. Five testers were involved in the data collection process. They were Sports Science graduating students doing their final year project and were supervised by experienced motor assessor. Inter- and intra-tester reliability checks using intra-class coefficient (ICC) were carried out using the videotaped data. Results: The inter-tester reliability between the five testers for the eight tasks ranged from rᵢcc = 0.705 to rᵢcc = 0.995. This suggests that the average agreement between them was considered good to excellent. With the exception of one tester who had rᵢcc = 0.687 for one of the eight tasks (i.e. zip-zap hopping), the intra-tester reliability within each tester ranged from rᵢcc = 0.728 to rᵢcc = 1.000, and this also suggested good to excellent consistency within testers. Discussion: The use of multiple testers with good intra-tester reliability for different test stations is feasible. This method allows several participants to be assessed concurrently at different test stations and saves overall data collection time. Therefore, it is recommended that the administering of MABC-2 with multiple testers should be extended to other age bands ensuring the feasibility of such method for other age bands.

Keywords: adolescents, MABC, motor assessment, motor skills, reliability

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Authors: Marianna Maiaru, Gregory M. Odegard

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During the processing of high-performance thermoset polymer matrix composites, chemical reactions occur during elevated pressure and temperature cycles, causing the constituent monomers to crosslink and form a molecular network that gradually can sustain stress. As the crosslinking process progresses, the material naturally experiences a gradual shrinkage due to the increase in covalent bonds in the network. Once the cured composite completes the cure cycle and is brought to room temperature, the thermal expansion mismatch of the fibers and matrix cause additional residual stresses to form. These compounded residual stresses can compromise the reliability of the composite material and affect the composite strength. Composite process modeling is greatly complicated by the multiscale nature of the composite architecture. At the molecular level, the degree of cure controls the local shrinkage and thermal-mechanical properties of the thermoset. At the microscopic level, the local fiber architecture and packing affect the magnitudes and locations of residual stress concentrations. At the macroscopic level, the layup sequence controls the nature of crack initiation and propagation due to residual stresses. The goal of this research is use molecular dynamics (MD) and finite element analysis (FEA) to predict the residual stresses in composite laminates and the corresponding effect on composite failure. MD is used to predict the polymer shrinkage and thermomechanical properties as a function of degree of cure. This information is used as input into FEA to predict the residual stresses on the microscopic level resulting from the complete cure process. Virtual testing is subsequently conducted to predict strength allowables. Experimental characterization is used to validate the modeling.

Keywords: molecular dynamics, finite element analysis, processing modeling, multiscale modeling

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Authors: Elif Tugce Aksun Tumerkan

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Although seafood is an important part of human diet and categorized highly traded food industry internationally, it is remain overlooked generally in the global food security aspect. Food product authentication is the main interest in the aim of both avoids commercial fraud and to consider the risks that might be harmful to human health safety. In recent years, with increasing consumer demand for regarding food content and it's transparency, there are some instrumental analyses emerging for determining food fraud depend on some analytical methodologies such as proteomic and metabolomics. While, fish and seafood consumed as fresh previously, within advanced technology, processed or packaged seafood consumption have increased. After processing or packaging seafood, morphological identification is impossible when some of the external features have been removed. The main fish and seafood quality-related issues are the authentications of seafood contents such as mislabelling products which may be contaminated and replacement partly or completely, by lower quality or cheaper ones. For all mentioned reasons, truthful consistent and easily applicable analytical methods are needed for assurance the correct labelling and verifying of seafood products. DNA-barcoding methods become popular robust that used in taxonomic research for endangered or cryptic species in recent years; they are used for determining food traceability also. In this review, when comparing the other proteomic and metabolic analysis, DNA-based methods are allowing a chance to identification all type of food even as raw, spiced and processed products. This privilege caused by DNA is a comparatively stable molecule than protein and other molecules. Furthermore showing variations in sequence based on different species and founding in all organisms, make DNA-based analysis more preferable. This review was performed to clarify the main advantages of using DNA-barcoding for determining seafood fraud among other techniques.

Keywords: DNA-barcoding, genetic analysis, food fraud, mislabelling, packaged seafood

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Authors: Debit Datta, Jeffrey J. Coleman, Scott A. Enebak, Lori G. Eckhardt

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Loblolly pine (Pinus taeda) is a leading productive timber species in the southeastern USA. Over the past three years, an emerging threat is expressed by successive needle defoliation followed by stunted growth and tree mortality in loblolly pine plantations. Considering economic significance, it has now become a rising concern among landowners, forest managers, and forest health state cooperators. However, the symptoms of the disease were perplexed somewhat with root disease(s) and recurrently attributed to invasive Phytophthora species due to the similarity of disease nature and devastation. Therefore, the study investigated the potential causal agent of this disease and characterized the fungi associated with loblolly pine needle defoliation in the southeastern USA. Besides, 70 trees were selected at seven long-term monitoring plots at Chatom, Alabama, to monitor and record the annual disease incidence and severity. Based on colony morphology and ITS-rDNA sequence data, a total of 28 species of fungi representing 17 families have been recovered from diseased loblolly pine needles. The native brown-spot pathogen, Lecanosticta acicola, was the species most frequently recovered from unhealthy loblolly pine needles in combination with some other common needle cast and rust pathogen(s). Identification was confirmed using morphological similarity and amplification of translation elongation factor 1-alpha gene region of interest. Tagged trees were consistently found chlorotic and defoliated from 2019 to 2020. The current emergence of the brown-spot pathogen causing loblolly pine mortality necessitates the investigation of the role of changing climatic conditions, which might be associated with increased pathogen pressure to loblolly pines in the southeastern USA.

Keywords: brown-spot needle blight, loblolly pine, needle defoliation, plantation forestry

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Authors: El Montassir Dahmane, Nadia Eladlani, Aziz Ouahrouch, Mohammed Rhazi, Moha Taourirte

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The present study was aimed to approximate the optimal conditions to chromium recovery from wastewater by nanoparticles and whiskers of chitosan. Chitosan with an average molecular weight of 63 kDa and a 96% deacetylation degree was prepared according to our previous study. Chromium recovery is influenced by different parameters. In our search, we determined the appropriate range of pH to form chitosan–Cr(III), nanoparticles Cr(III), and whiskers– Cr(III) complex. We studied also the influence of chromium concentration and the nature of chitosan-based materials on the complexation process. Our main aim is to approximate the optimal conditions to remove chromium(III) from the tanning bath, recuperated from tannery wastewater of Marrakech in Morocco. A Perkin Elmer optima 2000 Inductively Coupled Plasma- Optical Emission Spectrometer (ICP-OES), was used to determine the quantity of chromium persistent in tannery wastewater after complexation phenomenon. To the best of our knowledge, this is the first report interested in the optimal conditions for chromium recovery from wastewater by nanoparticles and whiskers of chitosan. From our research, we found that in chromium solution, the appropriate range of pH to form complex is between 5.6 and 6.7. Also, the complexation of Cr(III) is depending on the nature of complexing ligand and chromium concentration. The obtained results reveal that nanoparticles present an excellent adsorption capacity regardless of chromium concentration. In addition, after a critical chromium concentration (250 mg/l), our ligand becomes saturated, that requires an increase of ligand mass for increasing chromium concentration in order to have a better adsorption capacity. Hence, in the same conditions, we used chitosan, its nanoparticles, whiskers, and chitosan based films to remove Cr(III) from tannery wastewater. The pH of this effluent was around 6, and its chromium concentration was 300 mg/l. The results expose that the sequence of complexing ligand in the effluent is the same in chromium solution, determined via our previous study. However, the adsorbed quantity is less due to the presence of other metallic ions in tannery wastewater. We conclude that the best complexing ligand-based chitosan is chitosan nanoaprticles whether it’s in chromium solution or in tannery wastewater. Nanoparticles are the best complexing ligand after 24 h of contact nanoparticles can remove 70% of chromium from this tannery wastewater.

Keywords: nanoparticles, whiskers, chitosan, chromium

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Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E.histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

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Authors: Abhilash Sahu, Amit Priyadarshi

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In this paper, we will discuss the existence of weak solutions of the Kirchhoff type boundary value problem on the Sierpinski gasket. Where S denotes the Sierpinski gasket in R² and S₀ is the intrinsic boundary of the Sierpinski gasket. M: R → R is a positive function and h: S × R → R is a suitable function which is a part of our main equation. ∆p denotes the p-Laplacian, where p > 1. First of all, we will define a weak solution for our problem and then we will show the existence of at least two solutions for the above problem under suitable conditions. There is no well-known concept of a generalized derivative of a function on a fractal domain. Recently, the notion of differential operators such as the Laplacian and the p-Laplacian on fractal domains has been defined. We recall the result first then we will address the above problem. In view of literature, Laplacian and p-Laplacian equations are studied extensively on regular domains (open connected domains) in contrast to fractal domains. In fractal domains, people have studied Laplacian equations more than p-Laplacian probably because in that case, the corresponding function space is reflexive and many minimax theorems which work for regular domains is applicable there which is not the case for the p-Laplacian. This motivates us to study equations involving p-Laplacian on the Sierpinski gasket. Problems on fractal domains lead to nonlinear models such as reaction-diffusion equations on fractals, problems on elastic fractal media and fluid flow through fractal regions etc. We have studied the above p-Laplacian equations on the Sierpinski gasket using fibering map technique on the Nehari manifold. Many authors have studied the Laplacian and p-Laplacian equations on regular domains using this Nehari manifold technique. In general Euler functional associated with such a problem is Frechet or Gateaux differentiable. So, a critical point becomes a solution to the problem. Also, the function space they consider is reflexive and hence we can extract a weakly convergent subsequence from a bounded sequence. But in our case neither the Euler functional is differentiable nor the function space is known to be reflexive. Overcoming these issues we are still able to prove the existence of at least two solutions of the given equation.

Keywords: Euler functional, p-Laplacian, p-energy, Sierpinski gasket, weak solution

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Authors: Balakrishna Shetty

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Introduction: Stem Cell Imaging is a challenging field since the advent of Stem Cell treatment in humans. Series of research on tagging and tracking the stem cells has not been very effective. The present study is an effort by the authors to track the stem cells injected into calf muscles by Magnetic Resonance Diffusion Weighted Imaging. Materials and methods: Stem Cell injection deep into the calf muscles of patients with peripheral vascular disease is one of the recent treatment modalities followed in our institution. 5 patients who underwent deep intramuscular injection of stem cells as treatment were included for this study. Pre and two hours Post injection MRI of bilateral calf regions was done using 1.5 T Philips Achieva, 16 channel system using 16 channel torso coils. Axial STIR, Axial Diffusion weighted images with b=0 and b=1000 values with back ground suppression (DWIBS sequence of Philips MR Imaging Systems) were obtained at 5 mm interval covering the entire calf. The invert images were obtained for better visualization. 120ml of autologous bone marrow derived stem cells were processed and enriched under c-GMP conditions and reduced to 40ml solution containing mixture of above stem cells. Approximately 40 to 50 injections, each containing 0.75ml of processed stem cells, was injected with marked grids over the calf region. Around 40 injections, each of 1ml normal saline, is injected into contralateral leg as control. Results: Significant Diffusion hyper intensity is noted at the site of injected stem cells. No hyper intensity noted before the injection and also in the control side where saline was injected conclusion: This is one of the earliest studies in literature showing diffusion hyper intensity in intramuscularly injected stem cells. The advantages and deficiencies in this study will be discussed during the presentation.

Keywords: stem cells, imaging, DWI, peripheral vascular disease

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Authors: Mohammad Moridzadeh, Mohammad Djavid, Barry Doyle

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An assessment of the long-term stability of a cavern in Chagrin shale excavated by the sequential excavation method was performed during and after construction. During the excavation of the cavern, deformations of rock mass were measured at the surface of excavation and within the rock mass by surface and deep measurement instruments. Rock deformations were measured during construction which appeared to result from the as-built excavation sequence that had potentially disturbed the rock and its behavior. Also some additional time dependent rock deformations were observed during and post excavation. Several opinions have been expressed to explain this time dependent deformation including stress changes induced by excavation, strain softening (or creep) in the beddings with and without clay and creep of the shaley rock under compressive stresses. In order to analyze and replicate rock behavior observed during excavation, including current and post excavation elastic, plastic, and time dependent deformation, Finite Element Analysis (FEA) was performed. The analysis was also intended to estimate long term deformation of the rock mass around the excavation. Rock mass behavior including time dependent deformation was measured by means of rock surface convergence points, MPBXs, extended creep testing on the long anchors, and load history data from load cells attached to several long anchors. Direct creep testing of Chagrin Shale was performed on core samples from the wall of the Pump Room. Results of these measurements were used to calibrate the FEA of the excavation. These analyses incorporate time dependent constitutive modeling for the rock to evaluate the potential long term movement in the roof, walls, and invert of the cavern. The modeling was performed due to the concerns regarding the unanticipated behavior of the rock mass as well as the forecast of long term deformation and stability of rock around the excavation.

Keywords: Cavern, Chagrin shale, creep, finite element.

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Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

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Authors: Ram Sharan Mehta, Gayanandra Malla, Anita Gurung, Anu Aryal, Divya Labh, Hricha Neupane

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Objectives: Lack of resuscitation skills of nurses and doctors in basic life support (BLS) and advanced life support (ALS) has been identified as a contributing factor to poor outcomes of cardiac arrest victims. The objective of this study was to examine retention of life support measures (BLS/ALS) knowledge and skills of nurses following education intervention programme. Materials and Methods: Pre-experimental research design was used to conduct the study among the nurses working in medical units of B.P Koirala Institute of Health Sciences, where CPR is very commonly performed. Using convenient sampling technique total of 20 nurses agreed to participate and give consent were included in the study. The theoretical, demonstration and re-demonstration were arranged involving the trained doctors and nurses during the three hours educational session. Post-test was carried out after two week of education intervention programme. The 2010 BLS & ALS guidelines were used as guide for the study contents. The collected data were analyzed using SPSS-15 software. Results: It was found that there is significant increase in knowledge after education intervention in the components of life support measures (BLS/ALS) i.e. ratio of chest compression to ventilation in BLS (P=0.001), correct sequence of CPR (p <0.001), rate of chest compression in ALS (P=0.001), the depth of chest compression in adult CPR (p<0.001), and position of chest compression in CPR (P=0.016). Nurses were well appreciated the programme and request to continue in future for all the nurses. Conclusions: At recent BLS/ALS courses (2010), a significant number of nurses remain without any such training. Action is needed to ensure all nurses receive BLS training and practice this skill regularly in order to retain their knowledge.

Keywords: pre-experimental, basic and advance life support, nurses, sampling technique

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Authors: Ali Kadivar, Kaveh Niayesh

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The integration of green energy resources is a major focus, and the role of Medium Voltage Direct Current (MVDC) systems is exponentially expanding. However, the protection of MVDC systems against DC faults is a challenge that can have consequences on reliable and safe grid operation. This challenge reveals the need for MVDC circuit breakers (MVDC CB), which are in infancies of their improvement. Therefore will be a lack of MVDC CBs standards, including thresholds for acceptable power losses and operation speed. To establish a baseline for comparison purposes, a benchmark system for testing future MVDC CBs is vital. The literatures just give the timing sequence of each switch and the emphasis is on the topology, without in-depth study on the control algorithm of DCCB, as the circuit breaker control system is not yet systematic. A digital testing benchmark is designed for the Proof-of-concept of simulation studies using software models. It can validate studies based on real-time digital simulators and Transient Network Analyzer (TNA) models. The proposed experimental setup utilizes data accusation from the accurate sensors installed on the tested MVDC CB and through general purpose input/outputs (GPIO) from the microcontroller and PC Prototype studies in the laboratory-based models utilizing Hardware-in-the-Loop (HIL) equipment connected to real-time digital simulators is achieved. The improved control algorithm of the circuit breaker can reduce the peak fault current and avoid arc resignation, helping the coordination of DCCB in relay protection. Moreover, several research gaps are identified regarding case studies and evaluation approaches.

Keywords: DC circuit breaker, hardware-in-the-loop, real time digital simulation, testing benchmark

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Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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

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

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

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Authors: M. A. Barros, M. V. Barros

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The aim of this work is to analyze the discursive dynamics of preservice teachers during the implementation of a didactic sequence on topics of Quantum Mechanics for High School. Our research methodology was qualitative, case study type, in which we selected two prospective teachers on the Physics Teacher Training Course of the Sao Carlos Institute of Physics, at the University of Sao Paulo/Brazil. The set of modes of communication analyzed were the intentions and interventions of the teachers, the established communicative approach, the patterns and the contents of the interactions between teachers and students. Data were collected through video recording, interviews and questionnaires conducted before and after an 8 hour mini-course, which was offered to a group of 20 secondary students. As teaching strategy we used an active learning methodology, called: Peer Instruction. The episodes pointed out that both future teachers used interactive dialogic and authoritative communicative approaches to mediate the discussion between peers. In the interactive dialogic dimension the communication pattern was predominantly I-R-F (initiation-response-feedback), in which the future teachers assisted the students in the discussion by providing feedback to their initiations and contributing to the progress of the discussions between peers. Although the interactive dialogic dimension has been preferential during the use of the Peer Instruction method the authoritative communicative approach was also employed. In the authoritative dimension, future teachers used predominantly the type I-R-E (initiation-response-evaluation) communication pattern by asking the students several questions and leading them to the correct answer. Among the main implications the work contributes to the improvement of the practices of future teachers involved in applying active learning methodologies in classroom by identifying the types of communicative approaches and communication patterns used, as well as researches on curriculum innovation in physics in high school.

Keywords: curricular innovation, high school, physics teaching, discursive dynamics

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Authors: Priya Varma, Denis John McKeown

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Psychological time, unlike physical time, is believed to be ‘compressive’ in the sense that the mental representations of a series of events may be internally arranged with ever decreasing inter-event spacing (looking back from the most recently encoded event). If this is true, the record within immediate memory of recent events is severely temporally distorted. Although this notion of temporal distortion of the memory record is captured within some theoretical accounts of human forgetting, notably temporal distinctiveness accounts, the way in which the fundamental nature of the distortion underpins memory and forgetting broadly is barely recognised or at least directly investigated. Our intention here was to manipulate the spacing of items for recall in order to ‘reverse’ this supposed natural compression within the encoding of the items. In Experiment 1 three schedules of presentation (expanding, contracting and fixed irregular temporal spacing) were created using logarithmic spacing of the words for both free and serial recall conditions. The results of recall of lists of 7 words showed statistically significant benefits of temporal isolation, and more excitingly the contracting word series (which we may think of as reversing the natural compression within the mental representation of the word list) showed best performance. Experiment 2 tested for effects of active verbal rehearsal in the recall task; this reduced but did not remove the benefits of our temporal scheduling manipulation. Finally, a third experiment used the same design but with Chinese characters as memoranda, in a further attempt to subvert possible verbal maintenance of items. One change to the design here was to introduce a probe item following the sequence of items and record response times to this probe. Together the outcomes of the experiments broadly support the notion of temporal compression within immediate memory.

Keywords: memory, serial position curves, temporal isolation, temporal schedules

Procedia PDF Downloads 217

Authors: Tae-Hoon Kim, Kwon-Ha Yoon, Hong Young Jun, Ki-Jong Kim, Young Hwan Lee, Myeung Su Lee, Keum Ha Choi, Ki Jung Yun, Eun Young Cho, Yong-Yeon Jeong, Chung-Hwan Jun

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Purpose: To assess the changes of hepatic metabolite for differentiation between non-alcoholic steatohepatitis (NASH) and simple steatosis on proton magnetic resonance spectroscopy (1H-MRS) in both humans and animal model. Methods: The local institutional review board approved this study and subjects gave written informed consent. 1H-MRS measurements were performed on a localized voxel of the liver using a point-resolved spectroscopy (PRESS) sequence and hepatic metabolites of alanine (Ala), lactate/triglyceride (Lac/TG), and TG were analyzed in NASH, simple steatosis and control groups. The group difference was tested with the ANOVA and Tukey’s post-hoc tests, and diagnostic accuracy was tested by calculating the area under the receiver operating characteristics (ROC) curve. The associations between metabolic concentration and pathologic grades or non-alcoholic fatty liver disease(NAFLD) activity scores were assessed by the Pearson’s correlation. Results: Patient with NASH showed the elevated Ala(p<0.001), Lac/TG(p < 0.001), TG(p < 0.05) concentration when compared with patients who had simple steatosis and healthy controls. The NASH patients were higher levels in Ala(mean±SEM, 52.5±8.3 vs 2.0±0.9; p < 0.001), Lac/TG(824.0±168.2 vs 394.1±89.8; p < 0.05) than simple steatosis. The area under the ROC curve to distinguish NASH from simple steatosis was 1.00 (95% confidence interval; 1.00, 1.00) with Ala and 0.782 (95% confidence interval; 0.61, 0.96) with Lac/TG. The Ala and Lac/TG levels were well correlated with steatosis grade, lobular inflammation, and NAFLD activity scores. The metabolic changes in human were reproducible to a mice model induced by streptozotocin injection and a high-fat diet. Conclusion: 1H-MRS would be useful for differentiation of patients with NASH and simple hepatic steatosis.

Keywords: non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, 1H MR spectroscopy, hepatic metabolites

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Authors: Sharique Ahmed, Khushtar Anwar Salman

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Alpha-1-antitrypsin (AAT) is a major plasma serine protease inhibitor (SERPIN). Hereditary AAT deficiency is one of the common diseases in some part of the world. AAT is mainly produced in the liver and functions to protect the lung against proteolytic damage (e.g., from neutrophil elastase) acting as the major inhibitor for neutrophil elastase. α (1)-Antitrypsin (AAT) deficiency is an under recognized genetic condition that affects approximately 1 in 2,000 to 1 in 5,000 individuals and predisposes to liver disease and early-onset emphysema. Not only does α-1-antitrypsin deficiency lead to disabling syndrome of pulmonary emphysema, there are other disorders too which include ANCA (antineutrophilic cytoplasmic antibody) positive Wegener's granulomatosis, diffuse bronchiectasis, necrotizing panniculitis in α-1-antitrypsin phenotype (S), idiopathic pulmonary fibrosis and steroid dependent asthma. Augmentation therapy with alpha-1 antitrypsin (AAT) from human plasma has been available for specific treatment of emphysema due to AAT deficiency. Apart from this several observations have also suggested a role for endogenous suppressors of HIV-1, alpha-1 antitrypsin (AAT) has been identified to be one of those. In view of its varied important role in humans, serum from a mammalian source was chosen for the isolation and purification. Studies were performed on the homogeneous fraction. This study suggests that the buffalo serum α-1-antritrypsin has characteristics close to ovine, dog, horse and more importantly to human α-1-antritrypsin in terms of its hydrodynamic properties such as molecular weight, carbohydrate content, etc. The similarities in the hydrodynamic properties of buffalo serum α-1-antitrypsin with other sources of mammalian α-1-antitrypsin mean that it can be further studied and be a potential source for "augmentation therapy", as well as a source of AAT replacement therapy to raise serum levels above the protective threshold. Other parameters like the amino acid sequence, the effect of denaturants, and the thermolability or thermostability of the inhibitor will be the interesting basis of future studies on buffalo serum alpha-1 antitrypsin (AAT).

Keywords: α-1-antitrypsin, augmentation therapy , hydrodynamic properties, serine protease inhibitor

Procedia PDF Downloads 489

Authors: Yang Ye, Hongfei Qiu, Yaqi Li

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The planning and construction of traditional cities are oriented by cars, which leads to the problems of insufficient urban public space, fragmentation, and low utilization efficiency. With the development of driverless technology, the urban structure will change from the traditional single-core grid structure to the multi-core model. In terms of traffic organization, with the release of land for traffic facilities, public space will become more continuous and integrated with traffic space. In the context of driverless technology, urban public reconstruction is characterized by modularization and high efficiency, and its planning and layout features accord with points (service facilities), lines (smart lines), surfaces (activity centers). The public space of driverless urban roads will provide diversified urban public facilities and services. The intensive urban layout makes the commercial public space realize the functions of central activities and style display, respectively, in the interior (building atrium) and the exterior (building periphery). In addition to recreation function, urban green space can also utilize underground parking space to realize efficient dispatching of shared cars. The roads inside the residential community will be integrated into the urban landscape, providing conditions for the community public activity space with changing time sequence and improving the efficiency of space utilization. The intervention of driverless technology will change the thinking of traditional urban construction and turn it into a human-oriented one. As a result, urban public space will be richer, more connected, more efficient, and the urban space justice will be optimized. By summarizing the frontier research, this paper discusses the impact of unmanned driving on cities, especially urban public space, which is beneficial for landscape architects to cope with the future development and changes of the industry and provides a reference for the related research and practice.

Keywords: driverless, urban public space, construction strategy, urban design

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Authors: Othman E. Othman, Hassan R. Darwish, Amira M. Nowier

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Lactoferrin (LF) is a multifunctional protein involved in economically production traits like milk protein composition and skeletal structure in small ruminants including sheep and goat. So, LF gene - with its genetic polymorphisms associated with production traits - is considered a candidate genetic marker used in marker-assisted selection in goats. This study aimed to identify the different alleles and genotypes of this gene in three Egyptian goat breeds using PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing. Genomic DNA was extracted from 120 animals belonging to Barki, Zaraibi, and Damascus goat breeds. Using specific primers, PCR amplified 247-bp fragments from exon 2 of LF goat gene. The PCR products were subjected to Single-Strand Conformation Polymorphism (SSCP) technique. The results showed the presence of two genotypes GG and AG in the tested animals. The frequencies of both genotypes varied among the three tested breeds with the highest frequencies of GG genotype in all tested goat breeds. The sequence analysis of PCR products representing these two detected genotypes declared the presence of an SNP (single nucleotide polymorphisms) substitution (G/A) among G and A alleles of this gene. The association between different LF genotypes and milk composition as well as body measurement was estimated. The comparison showed that the animals possess AG genotypes are superior over those with GG genotypes for different parameters of milk protein compositions and skeletal structures. This finding declared that allele A of LF gene is considered the promising marker for the productive traits in goat. In conclusion, the Egyptian goat breeds will be needed to enhance their milk protein composition and growth trait parameters through the increasing of allele A frequency in their herds depending on the superior production traits of this allele in goats.

Keywords: lLactoferrin gene, PCR-SSCP, SNPs, Egyptian goat

Procedia PDF Downloads 155

Authors: K. S. Vimal, D. Bodhini, K. Ramya, N. Lakshmipriya, R. M. Anjana, V. Sudha, J. A. Lovegrove, V. Mohan, V. Radha

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Gene-lifestyle interaction studies have been carried out in various populations. However, to date there are no studies in an Asian Indian population. Hence, we examined whether lifestyle factors such as diet and physical activity modify the association between fat mass and obesity–associated (FTO) gene variants and obesity and type 2 diabetes (T2D) in an Asian Indian population. We studied 734 unrelated T2D and 884 normal glucose-tolerant (NGT) participants randomly selected from the Chennai Urban Rural Epidemiology Study (CURES) in Southern India. Obesity was defined according to the World Health Organization Asia Pacific Guidelines (non-obese, BMI < 25 kg/m2; obese, BMI ≥ 25 kg/m2). Six single nucleotide polymorphisms (SNPs) in the FTO gene (rs9940128, rs7193144, rs8050136, rs918031, rs1588413 and rs11076023) identified from recent genome-wide association studies for T2D were genotyped by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Dietary assessment was carried out using a validated food frequency questionnaire and physical activity was based upon the self-report. Interaction analyses were performed by including the interaction terms in the model. A joint likelihood ratio test of the main SNP effects and the SNP-diet/physical activity interaction effects was used in the linear regression analyses to maximize statistical power. Statistical analyses were performed using STATA version 13. There was a significant interaction between FTO SNP rs8050136 and carbohydrate energy percentage (Pinteraction=0.04) on obesity, where the ‘A’ allele carriers of the SNP rs8050136 had 2.46 times higher risk of obesity than those with ‘CC’ genotype (P=3.0x10-5) among individuals in the highest tertile of carbohydrate energy percentage. Furthermore, among those who had lower levels of physical activity, the ‘A’ allele carriers of the SNP rs8050136 had 1.89 times higher risk of obesity than those with ‘CC’ genotype (P=4.0x10-5). We also found a borderline interaction between SNP rs11076023 and carbohydrate energy percentage (Pinteraction=0.08) on T2D, where the ‘A’ allele carriers in the highest tertile of carbohydrate energy percentage, had 1.57 times higher risk of T2D than those with ‘TT’ genotype (P=0.002). There was also a significant interaction between SNP rs11076023 and physical activity (Pinteraction=0.03) on T2D. No further significant interactions between SNPs and macronutrient intake or physical activity on obesity and T2D were observed. In conclusion, this is the first study to provide evidence for a gene-diet and gene-physical activity interaction on obesity and T2D in an Asian Indian population. These findings suggest that the association between FTO gene variants and obesity and T2D is influenced by carbohydrate intake and physical activity levels. Greater understanding of how FTO gene influences obesity and T2D through dietary and exercise interventions will advance the development of behavioral intervention and personalised lifestyle strategies predicted to reduce the development of metabolic diseases in ‘A’ allele carriers of both SNPs in this Asian Indian population.

Keywords: dietary intake, FTO, obesity, physical activity, type 2 diabetes, Asian Indian.

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