Search results for: signaling pathway

Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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Authors: Aoxue Yang, Weili Tian, Haikun Liu

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

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

Procedia PDF Downloads 105

Authors: Ali Hamza

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Accurate segmentation of breast ultrasound images is of paramount importance in enhancing the diagnostic capabilities of breast cancer detection. This study presents an approach utilizing the U-Net architecture for segmenting breast ultrasound images aimed at improving the accuracy and reliability of mass identification within the breast tissue. The proposed method encompasses a multi-stage process. Initially, preprocessing techniques are employed to refine image quality and diminish noise interference. Subsequently, the U-Net architecture, a deep learning convolutional neural network (CNN), is employed for pixel-wise segmentation of regions of interest corresponding to potential breast masses. The U-Net's distinctive architecture, characterized by a contracting and expansive pathway, enables accurate boundary delineation and detailed feature extraction. To evaluate the effectiveness of the proposed approach, an extensive dataset of breast ultrasound images is employed, encompassing diverse cases. Quantitative performance metrics such as the Dice coefficient, Jaccard index, sensitivity, specificity, and Hausdorff distance are employed to comprehensively assess the segmentation accuracy. Comparative analyses against traditional segmentation methods showcase the superiority of the U-Net architecture in capturing intricate details and accurately segmenting breast masses. The outcomes of this study emphasize the potential of the U-Net-based segmentation approach in bolstering breast ultrasound image analysis. The method's ability to reliably pinpoint mass boundaries holds promise for aiding radiologists in precise diagnosis and treatment planning. However, further validation and integration within clinical workflows are necessary to ascertain their practical clinical utility and facilitate seamless adoption by healthcare professionals. In conclusion, leveraging the U-Net architecture for breast ultrasound image segmentation showcases a robust framework that can significantly enhance diagnostic accuracy and advance the field of breast cancer detection. This approach represents a pivotal step towards empowering medical professionals with a more potent tool for early and accurate breast cancer diagnosis.

Keywords: mage segmentation, U-Net, deep learning, breast cancer detection, diagnostic accuracy, mass identification, convolutional neural network

Procedia PDF Downloads 58

Authors: Afifa Hafidh, Fathi Touati, Ahmed Hichem Hamzaoui, Sayda Somrani

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The objective of the present study has been to synthesize and to characterize silica hybrid materials using sol-gel technic and to investigate their properties. Silica materials were successfully fabricated using various bi-functional 1,3,4-thiadiazoles and tetraethoxysilane (TEOS) as co-precursors via a facile one-pot sol-gel pathway. TEOS was introduced at room temperature with 1,3,4-thiadiazole 2,5-difunctiunal adducts, in ethanol as solvent and using HCl acid as catalyst. The sol-gel process lead to the formation of monolithic, coloured and transparent gels. TEOS was used as a principal network forming agent. The incorporation of 1,3,4-thiadiazole molecules was realized by attachment of these later onto a silica matrix. This allowed covalent linkage between organic and inorganic phases and lead to the formation of Si-N and Si-S bonds. The prepared hybrid materials were characterized by Fourier transform infrared, NMR ²⁹Si and ¹³C, scanning electron microscopy and nitrogen absorption-desorption measurements. The optic and magnetic properties of hybrids are studied respectively by ultra violet-visible spectroscopy and electron paramagnetic resonance. It was shown in this work, that heterocyclic moieties were successfully attached in the hybrid skeleton. The formation of the Si-network composed of cyclic units (Q3 structures) connected by oxygen bridges (Q4 structures) was proved by ²⁹Si NMR spectroscopy. The Brunauer-Elmet-Teller nitrogen adsorption-desorption method shows that all the prepared xerogels have isotherms type IV and are mesoporous solids. The specific surface area and pore volume of these materials are important. The obtained results show that all materials are paramagnetic semiconductors. The data obtained by Nuclear magnetic resonance ²⁹Si and Fourier transform infrared spectroscopy, show that Si-OH and Si-NH groups existing in silica hybrids can participate in adsorption interactions. The obtained materials containing reactive centers could exhibit adsorption properties of metal ions due to the presence of OH and NH functionality in the mesoporous frame work. Our design of a simple method to prepare hybrid materials may give interest of the development of mesoporous hybrid systems and their use within the domain of environment in the future.

Keywords: hybrid materials, sol-gel process, 1, 3, 4-thiadaizole, TEOS

Procedia PDF Downloads 152

Authors: Chen Shuwen, Nunthapin Chantachaimongkol

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Since the world has moved into a transitional period, known as globalization; the business environment is now more complicated than ever before. Corporate information has become a matter of great importance for stakeholders, in order to understand the current situation. As a result of this, the concept of corporate governance has been broadly introduced to manage and control the affairs of corporations while businesses are required to disclose both financial and non-financial information to public via various communication channels such as the annual report, the financial report, the company’s website, etc. However, currently there are several other issues related to asymmetric information such as moral hazard or adverse selection that still occur intensively in workplaces. To prevent such problems in the business, it is required to have an understanding of what factors strengthen their transparency, accountability, fairness, and responsibility. Under aforementioned arguments, this paper aims to propose a conceptual framework that enables an investigation on how corporate governance mechanism influences disclosure efficiency of listed companies in the Association of Southeast Asia Nations (ASEAN) and the factors that should be considered for further development of good behaviors, particularly in regards to voluntary disclosure practices. To achieve its purpose, extensive reviews of literature are applied as a research methodology. It is divided into three main steps. Firstly, the theories involved with both corporate governance and disclosure practices such as agency theory, contract theory, signaling theory, moral hazard theory, and information asymmetry theory are examined to provide theoretical backgrounds. Secondly, the relevant literatures based on multi- perspectives of corporate governance, its attributions and their roles on business processes, the influences of corporate governance mechanisms on business performance, and the factors determining corporate governance characteristics as well as capability are reviewed to outline the parameters that should be included in the proposed model. Thirdly, the well-known regulatory document OECD principles and previous empirical studies on the corporate disclosure procedures are evaluated to identify the similarities and differentiations with the disclosure patterns in the ASEAN. Following the processes and consequences of the literature review, abundant factors and variables are found. Further to the methodology, additional critical factors that also have an impact on the disclosure behaviors are addressed in two groups. In the first group, the factors which are linked to the national characteristics - the quality of national code, legal origin, culture, the level of economic development, and so forth. Whereas in the second group, the discoveries which refer to the firm’s characteristics - ownership concentration, ownership’s rights, controlling group, and so on. However, because of research limitations, only some literature are chosen and summarized to form part of the conceptual framework that explores the relationship between corporate governance and the disclosure practices of listed companies in ASEAN.

Keywords: corporate governance, disclosure practice, ASEAN, listed company

Procedia PDF Downloads 176

Authors: Elena Kokkoni, Prasanna Kannappan, Ashkan Zehfroosh, Effrosyni Mavroudi, Kristina Strother-Garcia, James C. Galloway, Jeffrey Heinz, Rene Vidal, Herbert G. Tanner

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Early immobility affects the motor, cognitive, and social development. Current pediatric rehabilitation lacks the technology that will provide the dosage needed to promote mobility for young children at risk. The addition of socially assistive robots in early interventions may help increase the mobility dosage. The aim of this study is to examine the feasibility of an early intervention paradigm where non-walking infants experience independent mobility while socially interacting with robots. A dynamic environment is developed where both the child and the robot interact and learn from each other. The environment involves: 1) a range of physical activities that are goal-oriented, age-appropriate, and ability-matched for the child to perform, 2) the automatic functions that perceive the child’s actions through novel activity recognition algorithms, and decide appropriate actions for the robot, and 3) a networked visual data acquisition system that enables real-time assessment and provides the means to connect child behavior with robot decision-making in real-time. The environment was tested by bringing a two-year old boy with Down syndrome for eight sessions. The child presented delays throughout his motor development with the current being on the acquisition of walking. During the sessions, the child performed physical activities that required complex motor actions (e.g. climbing an inclined platform and/or staircase). During these activities, a (wheeled or humanoid) robot was either performing the action or was at its end point 'signaling' for interaction. From these sessions, information was gathered to develop algorithms to automate the perception of activities which the robot bases its actions on. A Markov Decision Process (MDP) is used to model the intentions of the child. A 'smoothing' technique is used to help identify the model’s parameters which are a critical step when dealing with small data sets such in this paradigm. The child engaged in all activities and socially interacted with the robot across sessions. With time, the child’s mobility was increased, and the frequency and duration of complex and independent motor actions were also increased (e.g. taking independent steps). Simulation results on the combination of the MDP and smoothing support the use of this model in human-robot interaction. Smoothing facilitates learning MDP parameters from small data sets. This paradigm is feasible and provides an insight on how social interaction may elicit mobility actions suggesting a new early intervention paradigm for very young children with motor disabilities. Acknowledgment: This work has been supported by NIH under grant #5R01HD87133.

Keywords: activity recognition, human-robot interaction, machine learning, pediatric rehabilitation

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Authors: Regiane A. Oliveira, Carlos E. Vaz Rossell, Rubens Maciel Filho

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Lactic acid, besides a valuable chemical may be considered a platform for other chemicals. In fact, the feasibility of hemicellulosic sugars as feedstock for lactic acid production process, may represent the drop of some of the barriers for the second generation bioproducts, especially bearing in mind the 5-carbon sugars from the pre-treatment of sugarcane bagasse. Bearing this in mind, the purpose of this study was to use the hemicellulosic liquor from sugarcane bagasse as a substrate to produce lactic acid by fermentation. To release of sugars from hemicellulose it was made a pre-treatment with a diluted sulfuric acid in order to obtain a xylose's rich liquor with low concentration of inhibiting compounds for fermentation (≈ 67% of xylose, ≈ 21% of glucose, ≈ 10% of cellobiose and arabinose, and around 1% of inhibiting compounds as furfural, hydroxymethilfurfural and acetic acid). The hemicellulosic sugars associated with 20 g/L of yeast extract were used in a fermentation process with Lactobacillus plantarum to produce lactic acid. The fermentation process pH was controlled with automatic injection of Ca(OH)2 to keep pH at 6.00. The lactic acid concentration remained stable from the time when the glucose was depleted (48 hours of fermentation), with no further production. While lactic acid is produced occurs the concomitant consumption of xylose and glucose. The yield of fermentation was 0.933 g lactic acid /g sugars. Besides, it was not detected the presence of by-products, what allows considering that the microorganism uses a homolactic fermentation to produce its own energy using pentose-phosphate pathway. Through facultative heterofermentative metabolism the bacteria consume pentose, as is the case of L. plantarum, but the energy efficiency for the cell is lower than during the hexose consumption. This implies both in a slower cell growth, as in a reduction in lactic acid productivity compared with the use of hexose. Also, L. plantarum had shown to have a capacity for lactic acid production from hemicellulosic hydrolysate without detoxification, which is very attractive in terms of robustness for an industrial process. Xylose from hydrolyzed bagasse and without detoxification is consumed, although the hydrolyzed bagasse inhibitors (especially aromatic inhibitors) affect productivity and yield of lactic acid. The use of sugars and the lack of need for detoxification of the C5 liquor from sugarcane bagasse hydrolyzed is a crucial factor for the economic viability of second generation processes. Taking this information into account, the production of second generation lactic acid using sugars from hemicellulose appears to be a good alternative to the complete utilization of sugarcane plant, directing molasses and cellulosic carbohydrates to produce 2G-ethanol, and hemicellulosic carbohydrates to produce 2G-lactic acid.

Keywords: fermentation, lactic acid, hemicellulosic sugars, sugarcane

Procedia PDF Downloads 350

Authors: Aurea C. L. Lacerda, Paulo R. H. Moreno, Bruna M. P. Vianna, Cristina H. R. Serra, Airton Martin, André R. Baby, Vladi O. Consiglieri, Telma M. Kaneko

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The modified Franz cell is the most widely used model for in vitro permeation studies, however it still presents some disadvantages. Thus, some alternative methods have been developed such as Skin PAMPA, which is a bio- artificial membrane that has been applied for skin penetration estimation of xenobiotics based on HT permeability model consisting. Skin PAMPA greatest advantage is to carry out more tests, in a fast and inexpensive way. The membrane system mimics the stratum corneum characteristics, which is the primary skin barrier. The barrier properties are given by corneocytes embedded in a multilamellar lipid matrix. This layer is the main penetration route through the paracellular permeation pathway and it consists of a mixture of cholesterol, ceramides, and fatty acids as the dominant components. However, there is no consensus on the membrane composition. The objective of this work was to compare the performance among different bio-artificial membranes for studying the permeation in skin PAMPA system. Material and methods: In order to mimetize the lipid composition`s present in the human stratum corneum six membranes were developed. The membrane composition was equimolar mixture of cholesterol, ceramides 1-O-C18:1, C22, and C20, plus fatty acids C20 and C24. The membrane integrity assay was based on the transport of Brilliant Cresyl Blue, which has a low permeability; and Lucifer Yellow with very poor permeability and should effectively be completely rejected. The membrane characterization was performed using Confocal Laser Raman Spectroscopy, using stabilized laser at 785 nm with 10 second integration time and 2 accumulations. The membrane behaviour results on the PAMPA system were statistically evaluated and all of the compositions have shown integrity and permeability. The confocal Raman spectra were obtained in the region of 800-1200 cm-1 that is associated with the C-C stretches of the carbon scaffold from the stratum corneum lipids showed similar pattern for all the membranes. The ceramides, long chain fatty acids and cholesterol in equimolar ratio permitted to obtain lipid mixtures with self-organization capability, similar to that occurring into the stratum corneum. Conclusion: The artificial biological membranes studied for Skin PAMPA showed to be similar and with comparable properties to the stratum corneum.

Keywords: bio-artificial membranes, comparison, confocal Raman, skin PAMPA

Procedia PDF Downloads 483

Authors: Norihiro Kato, Yuriko Takayama

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

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

Procedia PDF Downloads 245

Authors: Lubaina Soni, Claire Farrell, Christopher Szota, Tim D. Fletcher

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Green roofs are a promising engineered ecosystem for reducing stormwater runoff and restoring vegetation cover in cities. Plants can contribute to rainfall retention by rapidly depleting water in the substrate; however, this increases the risk of plant drought stress. Green roof configurations, therefore, need to provide plants the opportunity to efficiently deplete the substrate but also avoid severe drought stress. This study used green roof modules placed in a rainout shelter during a six-month rainfall regime simulated in Melbourne, Australia. Rainfall was applied equally with an overhead irrigation system on each module. Aside from rainfall, modules were under natural climatic conditions, including temperature, wind, and radiation. A single species, Ficinia nodosa, was planted with five different treatments and three replicates of each treatment. In this experiment, we tested the impact of three plant cover treatments (0%, 50% and 100%) on rainfall retention and plant drought stress. We also installed two runoff zone treatments covering 50% of the substrate surface for additional modules with 0% and 50% plant cover to determine whether directing rainfall resources towards plant roots would reduce drought stress without impacting rainfall retention. The retention performance for the simulated rainfall events was measured, quantifying all components for hydrological performance and survival on green roofs. We found that evapotranspiration and rainfall retention were similar for modules with 50% and 100% plant cover. However, modules with 100% plant cover showed significantly higher plant drought stress. Therefore, planting at a lower cover/density reduced plant drought stress without jeopardizing rainfall retention performance. Installing runoff zones marginally reduced evapotranspiration and rainfall retention, but by approximately the same amount for modules with 0% and 50% plant cover. This indicates that reduced evaporation due to the installation of the runoff zones likely contributed to reduced evapotranspiration and rainfall retention. Further, runoff occurred from modules with runoff zones faster than those without, indicating that we created a faster pathway for water to enter and leave the substrate, which also likely contributed to lower overall evapotranspiration and retention. However, despite some loss in retention performance, modules with 50% plant cover installed with runoff zones showed significantly lower drought stress in plants compared to those without runoff zones. Overall, we suggest that reducing plant cover represents a simple means of optimizing green roof performance but creating runoff zones may reduce plant drought stress at the cost of reduced rainfall retention.

Keywords: green roof, plant cover, plant drought stress, rainfall retention

Procedia PDF Downloads 94

Authors: Hamza Javar Magnier, Robin Curtis

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There have been rapid advances in the upstream processing of protein therapeutics, which has shifted the bottleneck to downstream purification and formulation. Finding liquid formulations with shelf lives of up to two years is increasingly difficult for some of the newer therapeutics, which have been engineered for activity, but their formulations are often viscous, can phase separate, and have a high propensity for irreversible aggregation1. We explore means to develop improved predictive ability from a better understanding of how protein-protein interactions on formulation conditions (pH, ionic strength, buffer type, presence of excipients) and how these impact upon the initial steps in protein self-association and aggregation. In this work, we study the initial steps in the aggregation pathways using a minimal protein model based on square-well potentials and discontinuous molecular dynamics. The effect of model parameters, including range of interaction, stiffness, chain length, and chain sequence, implies that protein models fold according to various pathways. By reducing the range of interactions, the folding- and collapse- transition come together, and follow a single-step folding pathway from the denatured to the native state2. After parameterizing the model interaction-parameters, we developed an understanding of low-temperature conformational properties and fluctuations, and the correlation to the folding transition of proteins in isolation. The model fluctuations increase with temperature. We observe a low-temperature point, below which large fluctuations are frozen out. This implies that fluctuations at low-temperature can be correlated to the folding transition at the melting temperature. Because proteins “breath” at low temperatures, defining a native-state as a single structure with conserved contacts and a fixed three-dimensional structure is misleading. Rather, we introduce a new definition of a native-state ensemble based on our understanding of the core conservation, which takes into account the native fluctuations at low temperatures. This approach permits the study of a large range of length and time scales needed to link the molecular interactions to the macroscopically observed behaviour. In addition, these models studied are parameterized by fitting to experimentally observed protein-protein interactions characterized in terms of osmotic second virial coefficients.

Keywords: protein folding, native-ensemble, conformational fluctuation, aggregation

Procedia PDF Downloads 337

Authors: Pierre-Louis Lucas, Corinne Loutelier-Bourhis, Narimane Mati-Baouche, Philippe Chan Tchi-Song, Patrice Lerouge, Elodie Mathieu-Rivet, Muriel Bardor

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N-glycosylation is a post-translational modification taking place in the Endoplasmic Reticulum and the Golgi apparatus where defined glycan features are added on protein in a very specific sequence Asn-X-Thr/Ser/Cys were X can be any amino acid except proline. Because it is well-established that those N-glycans play a critical role in protein biological activity, protein half-life and that a different N-glycan structure may induce an immune response, they are very important in Biopharmaceuticals which are mainly glycoproteins bearing N-glycans. From now, most of the biopharmaceuticals are produced by mammalian cells like Chinese Hamster Ovary cells (CHO) for their N-glycosylation similar to the human, but due to the high production costs, several other species are investigated as the possible alternative system. In this purpose, the green microalgae Chlamydomonas reinhardtii was investigated as the potential production system for Biopharmaceuticals. This choice was influenced by the facts that C. reinhardtii is a well-study microalgae which is growing fast with a lot of molecular biology tools available. This organism is also producing N-glycan on its endogenous proteins. However, the analysis of the N-glycan structure of this microalgae has revealed some differences as compared to the human. Rather than in Human where the glycans are processed by key enzymes called N-acetylglucosaminyltransferase I and II (GnTI and GnTII) adding GlcNAc residue to form a GlcNAc₂Man₃GlcNAc₂ core N-glycan, C. reinhardtii lacks those two enzymes and possess a GnTI independent glycosylation pathway. Moreover, some enzymes like xylosyltransferases and methyltransferases not present in human are supposed to act on the glycans of C. reinhardtii. Furthermore, the recent structural study by mass spectrometry shows that the N-glycosylation precursor supposed to be conserved in almost all eukaryotic cells results in a linear Man₅GlcNAc₂ rather than a branched one in C. reinhardtii. In this work, we will discuss the new released MS information upon C. reinhardtii N-glycan structure and their impact on our attempt to modify the glycan in a Human manner. Two strategies will be discussed. The first one consisted in the study of Xylosyltransferase insertional mutants from the CLIP library in order to remove xyloses from the N-glycans. The second will go further in the humanization by transforming the microalgae with the exogenous gene from Toxoplasma gondii having an activity similar to GnTI and GnTII with the aim to synthesize GlcNAc₂Man₃GlcNAc₂ in C. reinhardtii.

Keywords: Chlamydomonas reinhardtii, N-glycosylation, glycosyltransferase, mass spectrometry, humanization

Procedia PDF Downloads 147

Authors: Surapong Vongvatcharanon, Fardeela Binalee, Wandee Udomuksorn, Ekkasit Kumarnsit, Uraporn Vongvatcharanon

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Hypogonadism is characterized by a decline in sex hormone levels, especially testosterone. It has been shown to be an important contributor to the decrease in muscle mass, muscle strength and performance, a condition known as sarcopenia. Preparations from Butea superba Roxb. (red Kwao Krua) have been reported to have androgenic properties. The active compounds are proposed to be flavonoids and flavonoid glycosides. Treatment with B. superba has been shown to improve erectile dysfunction in males. Parvalbumin (PV) is a relaxing factor and identified in fast twitch fibers. Alterations of the PV levels affects skeletal muscle functions. This study aimed to investigate the effects of orhchidectomy, testosterone replacement and different doses of Butea superba Roxb. on the structure, performance, levels of parvalbumin, parvalbumin and androgen receptor immunoreactivities in the extensor digitorum longus (EDL) and gastrocnemius muscles of orchidectomized rats. Twelve-week old male Wistar rats were randomly divided into 6 groups; sham-operated (SHAM), orchidectomized (BS-0), orchidectomized group that was treated with testosterone replacement of 6 µg/kg (TP) or an orchidectomized group that was treated with various doses of an extract from Butea superba Roxb.; 5 mg/kg (BS-5), 50 mg/kg (BS-50) and 500 mg/kg (BS-500) all for 90 days. The testosterone level, epididymis, seminal vesicle, prostate gland, vas deference weight, muscle fiber size, strength and endurance in both the EDL and gastrocnemius muscle were decreased in the BS-0 group but increased in the testosterone replacement group. Treatment with the B. superba Roxb. extract replacement group improved muscle fiber size, strength and endurance, but not total testosterone levels, or the epididymis, seminal vesicle, prostate gland, vas deference weight. Furthermore, the parvalbumin level, parvalbumin and androgen receptor immunoreactivities were reduced in the BS-0 group but increased in the testosterone replacement group and the B. superba Roxb. extract groups for both the EDL and gastrocnemius muscle. This study indicated that the reduction of testosterone level led to a decrease of the androgen receptor density resulting in a decline in the muscle mass and parvalbumin levels. The decrease of parvalbumin levels affected muscle performance. Testosterone replacement increased the androgen receptor density and led to an increase of muscle mass and parvalbumin levels. The increase in the parvalbumin levels may result in an improvement of muscle performance. This may explain one mechanism of testosterone on muscle mass and strength in the testosterone dependent sarcopenia. The B. superba Roxb. extract groups also had improved muscle mass, strength and endurance, parvalbumin level, parvalbumin and androgen immunoreactivities compared to the BS-O group . Butea superba Roxb. Extracts contains a flavonoid (3, 7, 3'-Trihydroxy-4'-methoxyflavone), flavonoiglycoside (3, 3'-dihydroxy-4'-methoxyflavone-7-O-β-D-glucopyranoside) and isoflavanolignans (butesuperins A and butesuperins B) all known to inhibit the cAMP phosphodiesterase enzyme. Therefore, cAMP signaling may have adaptive effects on skeletal muscle by increasing muscle mass, strength and endurance.

Keywords: Butea superba, parvalbumin, skeletal muscle, orchidectomy

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Authors: Thauane Silva, Gabrielle do Vale, Andre Ferreira, Marilda Siqueira, Thiago Moreno L. Souza, Milene D. Miranda

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The exposure of A(H1N1)pdm09-infected epithelial cells (HeLa) to HIV-1 viral particles, or its gp120, enhanced interferon-induced transmembrane protein (IFITM3) content, a viral restriction factor (RF), resulting in a decrease in influenza replication. The gp120 binds to CCR5 (R5) or CXCR4 (X4) cell receptors during HIV-1 infection. Then, it is possible that the endogenous ligands of these receptors also modulate the expression of IFITM3 and other cellular factors that restrict influenza virus replication. Thus, the aim of this study is to analyze the role of cellular receptors R5 and X4 in modulating RFs in order to inhibit the replication of the influenza virus. A549 cells were treated with 2x effective dose (ED50) of endogenous R5 or X4 receptor agonists, CCL3 (20 ng/ml), CCL4 (10 ng/ml), CCL5 (10 ng/ml) and CXCL12 (100 ng/mL) or exogenous agonists, gp120 Bal-R5, gp120 IIIB-X4 and its mutants (5 µg/mL). The interferon α (10 ng/mL) and oseltamivir (60 nM) were used as a control. After 24 h post agonists exposure, the cells were infected with virus influenza A(H3N2) at 2 MOI (multiplicity of infection) for 1 h. Then, 24 h post infection, the supernatant was harvested and, the viral titre was evaluated by qRT-PCR. To evaluate IFITM3 and SAM and HD domain containing deoxynucleoside triphosphate triphosphohydrolase 1 (SAMHD1) protein levels, A549 were exposed to agonists for 24 h, and the monolayer was lysed with Laemmli buffer for western blot (WB) assay or fixed for indirect immunofluorescence (IFI) assay. In addition to this, we analyzed other RFs modulation in A549, after 24 h post agonists exposure by customized RT² Profiler Polymerase Chain Reaction Array. We also performed a functional assay in which SAMHD1-knocked-down, by single-stranded RNA (siRNA), A549 cells were infected with A(H3N2). In addition, the cells were treated with guanosine to assess the regulatory role of dNTPs by SAMHD1. We found that R5 and X4 agonists inhibited influenza replication in 54 ± 9%. We observed a four-fold increase in SAMHD1 transcripts by RFs mRNA quantification panel. After 24 h post agonists exposure, we did not observe an increase in IFITM3 protein levels through WB or IFI assays, but we observed an upregulation up to three-fold in the protein content of SAMHD1, in A549 exposed to agonists. Besides this, influenza replication enhanced in 20% in cell cultures that SAMDH1 was knockdown. Guanosine treatment in cells exposed to R5 ligands further inhibited influenza virus replication, suggesting that the inhibitory mechanism may involve the activation of the SAMHD1 deoxynucleotide triphosphohydrolase activity. Thus, our data show for the first time a direct relationship of SAMHD1 and inhibition of influenza replication, and provides perspectives for new studies on the signaling modulation, through cellular receptors, to induce proteins of great importance in the control of relevant infections for public health.

Keywords: chemokine receptors, gp120, influenza, virus restriction factors

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Authors: Scott Hollier, Ruchi Permvattana

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The purpose of this abstract is to share the findings of a recently completed disability-related Internet of Things (IoT) project undertaken at Curtin University in Australia. The project focused on identifying how IoT could support people with disabilities with their educational outcomes. To achieve this, the research consisted of an analysis of current literature and interviews conducted with students with vision, hearing, mobility and print disabilities. While the research acknowledged the ability to collect data with IoT is now a fairly common occurrence, its benefits and applicability still need to be grounded back into real-world applications. Furthermore, it is important to consider if there are sections of our society that may benefit from these developments and if those benefits are being fully realised in a rush by large companies to achieve IoT dominance for their particular product or digital ecosystem. In this context, it is important to consider a group which, to our knowledge, has had little specific mainstream focus in the IoT area –people with disabilities. For people with disabilities, the ability for every device to interact with us and with each other has the potential to yield significant benefits. In terms of engagement, the arrival of smart appliances is already offering benefits such as the ability for a person in a wheelchair to give verbal commands to an IoT-enabled washing machine if the buttons are out of reach, or for a blind person to receive a notification on a smartphone when dinner has finished cooking in an IoT-enabled microwave. With clear benefits of IoT being identified for people with disabilities, it is important to also identify what implications there are for education. With higher education being a critical pathway for many people with disabilities in finding employment, the question as to whether such technologies can support the educational outcomes of people with disabilities was what ultimately led to this research project. This research will discuss several significant findings that have emerged from the research in relation to how consumer-based IoT can be used in the classroom to support the learning needs of students with disabilities, how industrial-based IoT sensors and actuators can be used to monitor and improve the real-time learning outcomes for the delivery of lectures and student engagement, and a proposed method for students to gain more control over their learning environment. The findings shared in this presentation are likely to have significant implications for the use of IoT in the classroom through the implementation of affordable and accessible IoT solutions and will provide guidance as to how policies can be developed as the implications of both benefits and risks continue to be considered by educators.

Keywords: disability, higher education, internet of things, students

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Authors: Nadia E. Christiana, Azzahra A. N. Ginting, Ardhito Nurcahya, Havisa P. Novira

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One of the quickest ways to do a short trip in urban areas is by walking, either individually, in couple or groups. Walkability nowadays becomes one of the parameters to measure the quality of an urban neighborhood. As a Central Business District and public transport transit hub, Dukuh Atas area becomes one of the highest numbers of commuters that pass by the area and interchange between transportation modes daily. Thus, as a public transport hub, a lot of investment should be focused to speed up the development of the area that would support urban transit activity between transportation modes, one of them is revitalizing pedestrian walkways. The purpose of this research is to formulate the visual design concept of 'Walkable City' based on the results of the observation and a series of rankings. To achieve this objective, it is necessary to accomplish several stages of the research that consists of (1) Identifying the system of pedestrian paths in Dukuh Atas area using descriptive qualitative method (2) Analyzing the sidewalk walkability rate according to the perception and the walkability satisfaction rate using the characteristics of pedestrians and non-pedestrians in Dukuh Atas area by using Global Walkability Index analysis and Multicriteria Satisfaction Analysis (3) Analyzing the factors that determine the integration of pedestrian walkways in Dukuh Atas area using descriptive qualitative method. The results achieved in this study is that the walkability level of Dukuh Atas corridor area is 44.45 where the value is included in the classification of 25-49, which is a bit of facility that can be reached by foot. Furthermore, based on the questionnaire, satisfaction rate of pedestrian walkway in Dukuh Atas area reached a number of 64%. It is concluded that commuters have not been fully satisfied with the condition of the sidewalk. Besides, the factors that influence the integration in Dukuh Atas area have been reasonable as it is supported by the utilization of land and modes such as KRL, Busway, and MRT. From the results of all analyzes conducted, the visual design and the application of the concept of walkable city along the pathway pedestrian corridor of Dukuh Atas area are formulated. Achievement of the results of this study amounted to 80% which needs to be done further review of the results of the analysis. The work of this research is expected to be a recommendation or input for the government in the development of pedestrian paths in maximizing the use of public transportation modes.

Keywords: design, global walkability index, mass rapid transit, walkable city

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Authors: Mark De Ste Croix, Jonathan Hughes, Francisco Ayala, Michal Lehnert

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There are well-known challenges to implementing injury prevention training for youth players but no data are available on the knowledge and understanding of deliverers of such programmes at grass root level. To increase adoption and adherence to such programmes coach knowledge and understanding of injury risk and prevention is essential. Therefore, the purpose of this study was to examine grass-root coaches knowledge and understanding of injury risk and prevention in youth players. 68 grass root coaches (18 females and 50 males) who were attending a one-day injury prevention workshop completed a modified validated questionnaire exploring knowledge and understanding of injury risk and prevention in youth players. Only 59% of coaches agreed that youth players are at a high risk of suffering an injury. There were high levels of agreement that injuries can have negative impacts on team performance (75%) and can cause physical problems in later life (85%), however only around half of coaches felt that injuries affect youth players current quality of life (59%). There was strong agreement that it is possible to prevent injuries in youth players (84%), but coaches were generally unaware of programs to help prevent injuries (84%), and only 9% used some form of injury prevention program. Despite this, nearly all coaches felt that their coaching could benefit from a greater understanding of growth and maturation (91%), injury prevention programmes (91%) and specific exercises (93%) for youth athletes. 17% of coaches rated their knowledge of injury prevention as good/very good at the start of the workshop and this increased to 94% at the end of the workshop. 62% of coaches identified their attitude towards injury prevention as indifferent at the start of the workshop compared with only 1% at the end. Only 14% of coaches at the start of the workshop were confident to deliver an injury prevention session but 83% stated they were confident by the end of the workshop. Finally, 98% of coaches felt that the workshop provided them with the confidence and the knowledge to deliver an injury prevention session and 98% suggested that they would implement injury prevention into their coaching. These data suggest that there is a lack of understanding of grass root coaches that children are a high-risk group for injuries, and that such injuries impact on current quality of life. Despite understanding that injuries can be prevented most grass root coaches do not have the knowledge to implement injury prevention into their coaching and very few do. There is a common consensus amongst these coaches that a greater understanding of such programmes will enhance their coaching. The injury prevention workshop appears to have increased the knowledge and changed the attitude of coaches towards injury prevention. All coaches felt that the workshop provided them with the tools to adopt, implement and deliver injury prevention in their coaching. These data highlight that there is a clear need for education regarding injury risk and prevention to be embedded within the coach education pathway, especially at grass root level.

Keywords: coach education, injury prevention, knowledge, and understanding, youth

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Authors: Emery Coppola Jr., Marwan Sadat, Il Kim, Diane Trube, Richard Kurisko

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Contamination sites like landfills often pose significant risks to receptors like surface water bodies. Surface water bodies are often a source of recreation, including fishing and swimming, which not only enhances their value but also serves as a direct exposure pathway to humans, increasing their need for protection from water quality degradation. In this paper, a case study presents the potential effects of leachate-impacted groundwater from a large closed sanitary landfill on the surface water quality of the nearby Raritan River, situated in New Jersey. The study, performed over a two year period, included in-depth field evaluation of both the groundwater and surface water systems, and was supplemented by computer modeling. The analysis required delineation of a representative average daily groundwater discharge from the Landfill shoreline into the large, highly tidal Raritan River, with a corresponding estimate of daily mass loading of potential contaminants of concern. The average daily groundwater discharge into the river was estimated from a high-resolution water level study and a 24-hour constant-rate aquifer pumping test. The significant tidal effects induced on groundwater levels during the aquifer pumping test were filtered out using an advanced algorithm, from which aquifer parameter values were estimated using conventional curve match techniques. The estimated hydraulic conductivity values obtained from individual observation wells closely agree with tidally-derived values for the same wells. Numerous models were developed and used to simulate groundwater contaminant transport and surface water quality impacts. MODFLOW with MT3DMS was used to simulate the transport of potential contaminants of concern from the down-gradient edge of the Landfill to the Raritan River shoreline. A surface water dispersion model based upon a bathymetric and flow study of the river was used to simulate the contaminant concentrations over space within the river. The modeling results helped demonstrate that because of natural attenuation, the Landfill does not have a measurable impact on the river, which was confirmed by an extensive surface water quality study.

Keywords: groundwater flow and contaminant transport modeling, groundwater/surface water interaction, landfill leachate, surface water quality modeling

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Authors: Marian Emmanuel Okon

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Introduction: The Niger Delta region of Nigeria has a vast reserve of oil and gas, which has globally positioned the nation as the sixth largest exporter of crude oil. Production rapidly rose following oil discovery. In most oil producing nations of the world, the wealth generated from oil production and export has propelled economic advancement, enabling the development of industries and other relevant infrastructures. Therefore, it can be assumed that majority of the oil resource such as Nigeria’s, has the potential to improve the health of the population via job creation and derived revenues. However, the health benefits of this economic development might be offset by the environmental consequences of oil exploitation and production. Objective: This research aims to evaluate the balance between the health benefits of oil-led economic development and harmful environmental consequences of crude oil exploitation in Nigeria. Study Design: A pathway has been designed to guide data search and this study. The model created will assess the relationship between oil-led economic development and population health development via job creation, improvement of education, development of infrastructure and other forms of development as well as through harmful environmental consequences from oil activities. Data/Emerging Findings: Diverse potentially suitable datasets which are at different geographical scales have been identified, obtained or applied for and the dataset from the World Bank has been the most thoroughly explored. This large dataset contains information that would enable the longitudinal assessment of both the health benefits and harms from oil exploitation in Nigeria as well as identify the disparities that exist between the communities, states and regions. However, these data do not extend far back enough in time to capture the start of crude oil production. Thus, it is possible that the maximum economic benefits and health harms could be missed. To deal with this shortcoming, the potential for a comparative study with countries like United Kingdom, Morocco and Cote D’ivoire has also been taken into consideration, so as to evaluate the differences between these countries as well as identify the areas of improvement in Nigeria’s environmental and health policies. Notwithstanding, these data have shown some differences in each country’s economic, environmental and health state over time as well as a corresponding summary statistics. Conclusion: In theory, the beneficial effects of oil exploitation to the health of the population may be substantial as large swaths of the ‘wider determinants’ of population heath are influenced by the wealth of a nation. However, if uncontrolled, the consequences from environmental pollution and degradation may outweigh these benefits. Thus, there is a need to address this, in order to improve environmental and population health in Nigeria.

Keywords: environmental pollution, health benefits, oil-led economic development, petroleum exploitation

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Authors: Aya Al Masri, Kamel Guerchouche, Youssef Laynaoui, Safoin Aktaou, Malorie Martin, Fouad Maaloul

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Introduction: Mobile C-arms move throughout operating rooms of the operating theater. Being designed to move between rooms, they are not equipped with relays to retrieve the exposition information and export it outside the room. Therefore, no light signaling is available outside the room to warn the X-ray emission for staff. Inadvertent exposition of staff outside the operating theater is a real problem for radiation protection. The French standard NFC 15-160 require that: (1) access to any room containing an X-ray emitting device must be controlled by a light signage so that it cannot be inadvertently crossed, and (2) setting up an emergency button to stop the X-ray emission. This study presents a system that we developed to meet these requirements and the results of its clinical test. Materials and methods: The system is composed of two communicating boxes: o The "DetectBox" is to be installed inside the operating theater. It identifies the various operation states of the C-arm by analyzing its power supply signal. The DetectBox communicates (in wireless mode) with the second box (AlertBox). o The "AlertBox" can operate in socket or battery mode and is to be installed outside the operating theater. It detects and reports the state of the C-arm by emitting a real time light signal. This latter can have three different colors: red when the C-arm is emitting X-rays, orange when it is powered on but does not emit X-rays, and green when it is powered off. The two boxes communicate on a radiofrequency link exclusively carried out in the ‘Industrial, Scientific and Medical (ISM)’ frequency bands and allows the coexistence of several on-site warning systems without communication conflicts (interference). Taking into account the complexity of performing electrical works in the operating theater (for reasons of hygiene and continuity of medical care), this system (having a size <10 cm²) works in complete safety without any intrusion in the mobile C-arm and does not require specific electrical installation work. The system is equipped with emergency button that stops X-ray emission. The system has been clinically tested. Results: The clinical test of the system shows that: it detects X-rays having both high and low energy (50 – 150 kVp), high and low photon flow (0.5 – 200 mA: even when emitted for a very short time (<1 ms)), Probability of false detection < 10-5, it operates under all acquisition modes (continuous, pulsed, fluoroscopy mode, image mode, subtraction and movie mode), it is compatible with all C-arm models and brands. We have also tested the communication between the two boxes (DetectBox and AlertBox) in several conditions: (1) Unleaded room, (2) leaded room, and (3) rooms with particular configuration (sas, great distances, concrete walls, 3 mm of lead). The result of these last tests was positive. Conclusion: This system is a reliable tool to alert the staff present outside the operating room for X-ray emission and insure their radiation protection.

Keywords: Clinical test, Inadvertent staff exposition, Light signage, Operating theater

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Authors: Selisha A. Sooklal, Phelelani Mpangase, Shaun Aron, Karl Rumbold

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Organically bound fluorine (C-F bond) is extremely rare in nature. Despite this, the first fluorinated secondary metabolite, fluoroacetate, was isolated from the plant Dichapetalum cymosum (commonly known as Gifblaar). However, the enzyme responsible for fluorination (fluorinase) in Gifblaar was never isolated and very little progress has been achieved in understanding this process in higher plants. Fluorinated compounds have vast applications in the pharmaceutical, agrochemical and fine chemicals industries. Consequently, an enzyme capable of catalysing a C-F bond has great potential as a biocatalyst in the industry considering that the field of fluorination is virtually synthetic. As with any biocatalyst, a range of these enzymes are required. Therefore, it is imperative to expand the exploration for novel fluorinases. This study aimed to gain molecular insights into secondary metabolite biosynthesis in Gifblaar using a high-throughput sequencing-based approach. Mechanical wounding studies were performed using Gifblaar leaf tissue in order to induce expression of the fluorinase. The transcriptome of the wounded and unwounded plant was then sequenced on the Illumina HiSeq platform. A total of 26.4 million short sequence reads were assembled into 77 845 transcripts using Trinity. Overall, 68.6 % of transcripts were annotated with gene identities using public databases (SwissProt, TrEMBL, GO, COG, Pfam, EC) with an E-value threshold of 1E-05. Sequences exhibited the greatest homology to the model plant, Arabidopsis thaliana (27 %). A total of 244 annotated transcripts were found to be differentially expressed between the wounded and unwounded plant. In addition, secondary metabolic pathways present in Gifblaar were successfully reconstructed using Pathway tools. Due to lack of genetic information for plant fluorinases, a transcript failed to be annotated as a fluorinating enzyme. Thus, a local database containing the 5 existing bacterial fluorinases was created. Fifteen transcripts having homology to partial regions of existing fluorinases were found. In efforts to obtain the full coding sequence of the Gifblaar fluorinase, primers were designed targeting the regions of homology and genome walking will be performed to amplify the unknown regions. This is the first genetic data available for Gifblaar. It has provided novel insights into the mechanisms of metabolite biosynthesis and will allow for the discovery of the first eukaryotic fluorinase.

Keywords: biocatalyst, fluorinase, gifblaar, transcriptome

Procedia PDF Downloads 251

Authors: Rosa S. Wong, Keith T.S. Tung, Winnie W. Y. Tso, King-Wa Fu, Nirmala Rao, Patrick Ip

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Electronic devices have become an essential part of our lives. Various reports have highlighted the alarming usage of electronic devices at early ages and its long-term developmental consequences. More sedentary screen time was associated with increased adiposity, worse cognitive and motor development, and psychosocial health. Apart from the problems caused by children’s own screen time, parents today are often paying less attention to their children due to hand-held device. Some anecdotes suggest that distracted parenting has negative impact on parent-child relationship. This study examined whether distracted parenting detrimentally affected parent-child activities which may, in turn, impair children’s psychosocial health. In 2018/19, we recruited a cohort of preschoolers from 32 local kindergartens in Tin Shui Wai and Sham Shui Po for a 5-year programme aiming to build stronger foundations for children from disadvantaged backgrounds through an integrated support model involving medical, education and social service sectors. A comprehensive set of questionnaires were used to survey parents on their frequency of being distracted while parenting and their frequency of learning and recreational activities with children. Furthermore, they were asked to report children’s screen time amount and their psychosocial problems. Mediation analyses were performed to test the direct and indirect effects of electronic device-distracted parenting on children’s psychosocial problems. This study recruited 873 children (448 females and 425 males, average age: 3.42±0.35). Longer screen time was associated with more psychosocial difficulties (Adjusted B=0.37, 95%CI: 0.12 to 0.62, p=0.004). Children’s screen time positively correlated with electronic device-distracted parenting (r=0.369, p < 01). We also found that electronic device-distracted parenting was associated with more hyperactive/inattentive problems (Adjusted B=0.66, p < 0.01), fewer prosocial behavior (Adjusted B=-0.74, p < 0.01), and more emotional symptoms (Adjusted B=0.61, p < 0.001) in children. Further analyses showed that electronic device-distracted parenting exerted influences both directly and indirectly through parent-child interactions but to different extent depending upon the outcome under investigation (38.8% for hyperactivity/inattention, 31.3% for prosocial behavior, and 15.6% for emotional symptoms). We found that parents’ use of devices and children’s own screen time both have negative effects on children’s psychosocial health. It is important for parents to set “device-free times” each day so as to ensure enough relaxed downtime for connecting with children and responding to their needs.

Keywords: early childhood, electronic device, psychosocial wellbeing, parenting

Procedia PDF Downloads 139

Authors: Peter C. St. John, Michael F. Crowley, Yannick J. Bomble

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Production of chemicals from engineered organisms in a batch culture typically involves a trade-off between productivity, yield, and titer. However, strategies for strain design typically involve designing mutations to achieve the highest yield possible while maintaining growth viability. Such approaches tend to follow the principle of designing static networks with minimum metabolic functionality to achieve desired yields. While these methods are computationally tractable, optimum productivity is likely achieved by a dynamic strategy, in which intracellular fluxes change their distribution over time. One can use multi-stage fermentations to increase either productivity or yield. Such strategies would range from simple manipulations (aerobic growth phase, anaerobic production phase), to more complex genetic toggle switches. Additionally, some computational methods can also be developed to aid in optimizing two-stage fermentation systems. One can assume an initial control strategy (i.e., a single reaction target) in maximizing productivity - but it is unclear how close this productivity would come to a global optimum. The calculation of maximum theoretical yield in metabolic engineering can help guide strain and pathway selection for static strain design efforts. Here, we present a method for the calculation of a maximum theoretical productivity of a batch culture system. This method follows the traditional assumptions of dynamic flux balance analysis: that internal metabolite fluxes are governed by a pseudo-steady state and external metabolite fluxes are represented by dynamic system including Michealis-Menten or hill-type regulation. The productivity optimization is achieved via dynamic programming, and accounts explicitly for an arbitrary number of fermentation stages and flux variable changes. We have applied our method to succinate production in two common microbial hosts: E. coli and A. succinogenes. The method can be further extended to calculate the complete productivity versus yield Pareto surface. Our results demonstrate that nearly optimal yields and productivities can indeed be achieved with only two discrete flux stages.

Keywords: A. succinogenes, E. coli, metabolic engineering, metabolite fluxes, multi-stage fermentations, succinate

Procedia PDF Downloads 190

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: Jong H. Kim, Kathleen L. Chan, Luisa W. Cheng

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Control of fungal pathogens, such as foodborne mycotoxin producers, is problematic as effective antimycotic agents are often very limited. Mycotoxin contamination significantly interferes with the safe production of foods or crops worldwide. Moreover, expansion of fungal resistance to commercial drugs or fungicides is a global human health concern. Therefore, there is a persistent need to enhance the efficacy of commercial antimycotic agents or to develop new intervention strategies. Disruption of the cellular antioxidant system should be an effective method for pathogen control. Such disruption can be achieved with safe, redox-active compounds. Natural phenolic derivatives are potent redox cyclers that inhibit fungal growth through destabilization of the cellular antioxidant system. The goal of this study is to identify novel, redox-active compounds that disrupt the fungal antioxidant system. The identified compounds could also function as sensitizing agents to conventional antimycotics (i.e., chemosensitization) to improve antifungal efficacy. Various benzo derivatives were tested against fungal pathogens. Gene deletion mutants of the yeast Saccharomyces cerevisiae were used as model systems for identifying molecular targets of benzo analogs. The efficacy of identified compounds as potent antifungal agents or as chemosensitizing agents to commercial drugs or fungicides was examined with methods outlined by the Clinical Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing. Selected benzo derivatives possessed potent antifungal or antimycotoxigenic activity. Molecular analyses by using S. cerevisiae mutants indicated antifungal activity of benzo derivatives was through disruption of cellular antioxidant or cell wall integrity system. Certain benzo analogs screened overcame tolerance of Aspergillus signaling mutants, namely mitogen-activated protein kinase mutants, to fludioxonil fungicide. Synergistic antifungal chemosensitization greatly lowered minimum inhibitory or fungicidal concentrations of test compounds, including inhibitors of mitochondrial respiration. Of note, salicylaldehyde is a potent antimycotic volatile that has some practical application as a fumigant. Altogether, benzo derivatives targeting cellular antioxidant system of fungi (along with cell wall integrity system) effectively suppress fungal growth. Candidate compounds possess the antifungal, antimycotoxigenic or chemosensitizing capacity to augment the efficacy of commercial antifungals. Therefore, chemogenetic approaches can lead to the development of novel antifungal intervention strategies, which enhance the efficacy of established microbe intervention practices and overcome drug/fungicide resistance. Chemosensitization further reduces costs and alleviates negative side effects associated with current antifungal treatments.

Keywords: antifungals, antioxidant system, benzo derivatives, chemosensitization

Procedia PDF Downloads 229

Authors: Wei-Ju Huang, Hung-Pin Hsu

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[Background] In congestive heart failure (CHF), it has always been the principle of clinical treatment to control the water retention mechanism in the body to prevent excessive fluid retention. Early control of sympathetic nerves, Renin-Angiotensin-Aldosterone system (RAA system, RAAS), or strengthening of Atrial Natriuretic Peptide (ANP) was the point. In RAA system, related hormones, such as angiotensin, or enzymes in the pathway, such as ACE-I, can be used with corresponding inhibitors to reduce water content.[Aim] In recent years, clinical studies have pointed out that if different mechanisms are combined, the control effect seems to be better. For example, recent studies showed that ENTRESTO, a combination of Sacubitril and Valsartan, is a good new drug for CHF. Sacubitril is a prodrug. After activation, it can inhibit neprilysin and act as a neprilysin inhibitor (ARNI) to reduce the breakdown of natriuretic peptides(ANP). Valsartan is a kind of angiotensin receptor blocker (ARB), both of which are used to treat heart failure at the same time, have excellent curative effects.[Materials and Methods] Considering the side effects of this drug, coughing and a few cases of diarrhea were observed. However, the effect of this drug on the patient's intestinal tract has not been confirmed. On the other hand, studies have pointed out that ANP supplement can improve the CHF and increase the inhibitory effect on cancer cells. Therefore, the purpose of this study is to use a special microbial detection method to prove that whether oral drugs have an effect on microorganisms.The experimental method uses Nissui Compact Dry to observe the situation in different types of microorganisms. After the drug is dissolved in water, it is implanted in a petri dish, and the presence of different microorganisms is detected through different antibody reactions to confirm whether the drug has some toxicology in the gut.[Results and Discussion]From the above experimental results, it can be known that among the effects of Sacubitril and Valsartan on the basic microbial flora of the human body, low doses had no significant effect on Escherichia coli or intestinal bacteria. If Sacubitril or Valsartan with a high concentration of 3mg/ml is used alone or under the stimulation of a high concentration of the two drugs, it has a significant inhibitory effect on Escherichia coli. However, in terms of the effect on intestinal bacteria, high concentration of Sacubitril has a more significant inhibitory effect on intestinal bacteria, while high concentration of Valsartan has a less significant inhibitory effect on intestinal bacteria. The inhibitory effect of the combination of the two drugs on intestinal bacteria is also less significant.[Conclusion]The results of this study can be used as a further reference for the possible side effects of the clinical use of Sacubitril and Valsartan on the intestinal tract of patients,

Keywords: sacubitril, valsartan, entresto, congestive heart failure (CHF)

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Authors: Anupalli Roja Rani, Pavithra Dasari

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Background: Among several, breast cancer has emerged as the most common female cancer in developing countries. It is the most common cause of cancer-related deaths worldwide among women. It is a molecularly and clinically heterogeneous disease. Moreover, it is a hormone–dependent tumor in which estrogens can regulate the growth of breast cells by binding with estrogen receptors (ERs). Moreover, the use of natural products in cancer therapeutics is due to their properties of biocompatibility and less toxicity. Plants are the vast reservoirs for various bioactive compounds. Coleus barbatus (Lamiaceae) contains anticancer properties against several cancer cell lines. Method: In the present study, an attempt is being made to enrich the knowledge of the anticancer activity of pure compounds extracted from Coleus barbatus (Andrew). On human breast cancer cell lines MCF-7. Here in, we are assessing the antiproliferative activity of Coleus barbatus (Andrew) plant extracts against MCF 7 and also evaluating their toxicity in normal human mammary cell lines such as Human Mammary Epithelial Cells (HMEC). The active fraction of plant extract was further purified with the help of Flash chromatography, Medium Pressure Liquid Chromatography (MPLC) and preparative High-Performance Liquid Chromatography (HPLC). The structure of pure compounds will be elucidated by using modern spectroscopic methods like Nuclear magnetic resonance (NMR), Electrospray Ionisation Mass Spectrometry (ESI-MS) methods. Later, the growth inhibition morphological assessment of cancer cells and cell cycle analysis of purified compounds were assessed using FACS. The growth and progression of signaling molecules HER2, GRP78 was studied by secretion assay using ELISA and expression analysis by flow cytometry. Result: Cytotoxic effect against MCF-7 with IC50 values were derived from dose response curves, using six concentrations of twofold serially diluted samples, by SOFTMax Pro software (Molecular device) and respectively Ellipticine and 0.5% DMSO were used as a positive and negative control. Conclusion: The present study shows the significance of various bioactive compounds extracted from Coleus barbatus (Andrew) root material. It acts as an anti-proliferative and shows cytotoxic effects on human breast cancer cell lines MCF7. The plant extracts play an important role pharmacologically. The whole plant has been used in traditional medicine for decades and the studies done have authenticated the practice. Earlier, as described, the plant has been used in the ayurveda and homeopathy medicine. However, more clinical and pathological studies must be conducted to investigate the unexploited potential of the plant. These studies will be very useful for drug designing in the future.

Keywords: coleus barbatus, HPLC, MPLC, NMR, MCF7, flash chromatograph, ESI-MS, FACS, ELISA.

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Authors: Yu-Shun Elisa Pong

Abstract:

Music aesthetics related to phenomenology is rarely discussed and still in the ascendant while multi-dimensional discourses of philosophy were emerged to be an important trend in the 20th century. In the present study, a basic theory of phenomenology from Edmund Husserl (1859-1938) is revealed and discussed followed by the introduction of intentionality concepts, eidetic reduction, horizon, world, and inter-subjectivity issues. Further, phenomenology of music and general art was brought to attention by the introduction of Roman Ingarden’s The Work of Music and the Problems of its Identity (1933) and Mikel Dufrenne’s The Phenomenology of Aesthetic Experience (1953). Finally, Debussy’s music will be analyzed and discussed from the perspective of phenomenology. Phenomenology is not so much a methodology or analytics rather than a common belief. That is, as much as possible to describe in detail the different human experience, relative to the object of purpose. Such idea has been practiced in various guises for centuries, only till the early 20th century Phenomenology was better refined through the works of Husserl, Heidegger, Sartre, Merleau-Ponty and others. Debussy was born in an age when the Western society began to accept the multi-cultural baptism. With his unusual sensitivity to the oriental culture, Debussy has presented considerable inspiration, absorption, and echo in his music works. In fact, his relationship with nature is far from echoing the idea of Chinese ancient literati and nature. Although he is not the first composer to associate music with human and nature, the unique quality and impact of his works enable him to become a significant figure in music aesthetics. Debussy’s music tried to develop a quality analogous of nature, and more importantly, based on vivid life experience and artistic transformation to achieve the realm of pure art. Such idea that life experience comes before artwork, either clear or vague, simple or complex, was later presented abstractly in his late works is still an interesting subject worth further discussion. Debussy’s music has existed for more than or close to a century. It has received musicology researcher’s attention as much as other important works in the history of Western music. Among the pluralistic discussion about Debussy’s art and ideas, phenomenological aesthetics has enlightened new ideas and view angles to relook his great works and even gave some previous arguments legitimacy. Overall, this article provides a new insight of Debussy’s music from phenomenological exploration and it is believed phenomenology would be an important pathway in the research of the music aesthetics.

Keywords: Debussy's music, music esthetics, oriental culture, phenomenology

Procedia PDF Downloads 239

Authors: Jineetkumar Gawad, Chandrakant Bonde

Abstract:

Tuberculosis (TB) is a major worldwide concern whose control has been exacerbated by HIV, the rise of multidrug-resistance (MDR-TB) and extensively drug resistance (XDR-TB) strains of Mycobacterium tuberculosis. The interest for newer and faster acting antitubercular drugs are more remarkable than any time. To search potent compounds is need and challenge for researchers. Here, we tried to design lead for inhibition of Decaprenyl phosphoryl-β-D-ribose 2′-epimerase (DprE1) enzyme. Arabinose is an essential constituent of mycobacterial cell wall. DprE1 is a flavoenzyme that converts decaprenylphosphoryl-D-ribose into decaprenylphosphoryl-2-keto-ribose, which is intermediate in biosynthetic pathway of arabinose. Latter, DprE2 converts keto-ribose into decaprenylphosphoryl-D-arabinose. We had a selection of 23 compounds from azaindole series for computational study, and they were drawn using marvisketch. Ligands were prepared using Maestro molecular modeling interface, Schrodinger, v10.5. Common pharmacophore hypotheses were developed by applying dataset thresholds to yield active and inactive set of compounds. There were 326 hypotheses were developed. On the basis of survival score, ADRRR (Survival Score: 5.453) was selected. Selected pharmacophore hypotheses were subjected to virtual screening results into 1000 hits. Hits were prepared and docked with protein 4KW5 (oxydoreductase inhibitor) was downloaded in .pdb format from RCSB Protein Data Bank. Protein was prepared using protein preparation wizard. Protein was preprocessed, the workspace was analyzed using force field OPLS 2005. Glide grid was generated by picking single atom in molecule. Prepared ligands were docked with prepared protein 4KW5 using Glide docking. After docking, on the basis of glide score top-five compounds were selected, (5223, 5812, 0661, 0662, and 2945) and the glide docking score (-8.928, -8.534, -8.412, -8.411, -8.351) respectively. There were interactions of ligand and protein, specifically HIS 132, LYS 418, TRY 230, ASN 385. Pi-pi stacking was observed in few compounds with basic Imidazo [4,5-b] pyridine ring. We had basic azaindole ring in parent compounds, but after glide docking, we received compounds with Imidazo [4,5-b] pyridine as a basic ring. That might be the new lead in the process of drug discovery.

Keywords: DprE1 inhibitors, in silico drug designing, imidazo [4, 5-b] pyridine, lead, tuberculosis

Procedia PDF Downloads 129

Authors: Yanmei Xiong, Baowen Liao, Kun Xin, Zhongmao Jiang, Hao Guo, Yujun Chen, Mei Li

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Conversion of mangroves to aquaculture ponds represented as one major reason for mangrove loss in Asian countries in the 20th century. Recently the Chinese government has set a goal to increase 48,650 ha (more than the current mangrove area) of mangroves before the year of 2025 and “rehabilitation of aquaculture ponds to mangroves” projects are considered to be the major pathway to increase the mangrove area of China. It remains unclear whether natural colonization is feasible and what are the main influencing factors for mangrove restoration in these projects. In this study, a total of 17 rehabilitation sites in Dongzhai Bay, Hainan, China were surveyed for vegetation, soil and surface elevation five years after the rehabilitation project was initiated. Colonization of non-planted mangrove species was found at all sites and non-planted species dominated over planted species at 14 sites. Mangrove plants could only be found within the elevation range of -20 cm to 65 cm relative to the mean sea level. Soil carbon and nitrogen contents of the top 20 cm were generally low, ranging between 0.2%–1.4% and 0.03%–0.09%, respectively, and at each site, soil carbon and nitrogen were significantly lower at elevations with mangrove plants than lower elevations without mangrove plants. Seven sites located at the upper stream of river estuaries, where soil salinity was relatively lower, and nutrient was relatively higher, was dominated by non-planted Sonneratia caseolaris. Seven sites located at the down-stream of river estuaries or in the inner part of the bay, where soil salinity and nutrient were intermediate, were dominated by non-planted alien Sonneratia apetala. Another three sites located at the outer part of the bay, where soil salinity was higher and nutrient was lower, were dominated by planted species (Rhizophora stylosa, Kandelia obovata, Aegiceras corniculatum and Bruguiera sexangula) with non-planted S. apetala and Avicennia marina also found. The results suggest that natural colonization of mangroves is feasible in pond rehabilitation projects given the rehabilitation of tidal activities and appropriate elevations. Surface elevation is the major determinate for the success of mangrove rehabilitation, and soil salinity and nutrients are important in shaping vegetation structure. The colonization and dominance of alien species (Sonneratia apetala in this case) in some rehabilitation sites poses invasion risks and thus cautions should be taken when introducing alien mangrove species.

Keywords: coastal wetlands, ecological restoration, mangroves, natural colonization, shrimp pond rehabilitation, wetland restoration

Procedia PDF Downloads 110