Search results for: tissue damage
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3796

Search results for: tissue damage

346 A Left Testicular Cancer with Multiple Metastases Nursing Experience

Authors: Syue-Wen Lin

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Objective:This article reviews the care experience of a 40-year-old male patient who underwent a thoracoscopic right lower lobectomy following a COVID-19 infection. His complex medical history included multiple metastases (lungs, liver, spleen, and left kidney) and lung damage from COVID-19, which complicated the weaning process from mechanical ventilation. The care involved managing cancer treatment, postoperative pain, wound care, and palliative care. Methods:Nursing care was provided from August 16 to August 17, 2024. Challenges included difficulty with sputum clearance, which exacerbated the patient's anxiety and fear of reintubation. Pain management strategies combined analgesic drugs, non-drug methods, essential oil massages with family members, and playing the patient’s favorite music to reduce pain and anxiety. Progressive rehabilitation began with stabilizing vital signs, followed by assistance with sitting on the edge of the bed and walking within the ward. Strict sterile procedures and advanced wound care technology were used for daily dressing changes, with meticulous documentation of wound conditions and appropriate dressing selection. Holistic cancer care and palliative measures were integrated to address the patient’s physical and psychological needs. Results:The interdisciplinary care team developed a comprehensive plan addressing both physical and psychological aspects. Respiratory therapy, lung expansion exercises, and a high-frequency chest wall oscillation vest facilitated sputum expulsion and assisted in weaning from mechanical ventilation. The integration of cancer care, pain management, wound care, and palliative care led to improved quality of life and recovery. The collaborative approach between nursing staff and family ensured that the patient received compassionate and effective care. Conclusion: The complex interplay of emergency surgery, COVID-19, and advanced cancer required a multifaceted care strategy. The care team’s approach, combining critical care with tailored cancer and palliative care, effectively improved the patient’s quality of life and facilitated recovery. The comprehensive care plan, developed with family collaboration, provided both high-quality medical care and compassionate support for the terminally ill patient.

Keywords: multiple metastases, testicular cancer, palliative care, nursing experience

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345 The Impact of Mycotoxins on the Anaerobic Digestion Process

Authors: Harald Lindorfer, Bettina Frauz, Dietmar Ramhold

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Next to the well-known inhibitors in anaerobic digestion like ammonia, antibiotics or disinfectants, the number of process failures connected with mould growth in the feedstock increased significantly in the last years. It was assumed that mycotoxins are the cause of the negative effects. The financial damage to plants associated with these process failures is considerable. The aim of this study was to find a way of predicting the failures and furthermore strategies for a fast process recovery. In a first step, mould-contaminated feedstocks causing process failures in full-scale digesters were sampled and analysed on mycotoxin content. A selection of these samples was applied to biological inhibition tests. In this test, crystalline cellulose is applied in addition to the feedstock sample as standard substrate. Affected digesters were also sampled and analytical process data as well as operational data of the plants were recorded. Additionally, different mycotoxin substances, Deoxynivalenol, Zearalenon, Aflatoxin B1, Mycophenolic acid and Citrinin, were applied as pure substances to lab-scale digesters, individually and in various combinations, and effects were monitored. As expected, various mycotoxins were detected in all of the mould-contaminated samples. Nevertheless, inhibition effects were observed with only one of the collected samples, after applying it to an inhibition test. With this sample, the biogas yield of the standard substrate was reduced by approx. 20%. This result corresponds with observations made on full-scale plants. However, none of the tested mycotoxins applied as pure substance caused a negative effect on biogas production in lab scale digesters, neither after application as individual substance nor in combination. The recording of the process data in full-scale plants affected by process failures in most cases showed a severe accumulation of fatty acids alongside a decrease in biogas production and methane concentration. In the analytical data of the digester samples, a typical distribution of fatty acids with exceptionally high acetic acid concentrations could be identified. This typical fatty acid pattern can be used as a rapid identification parameter pointing to the cause of the process troubles and enable a fast implication of countermeasures. The results of the study show that more attention needs to be paid to feedstock storage and feedstock conservation before their application to anaerobic digesters. This is all the more important since first studies indicate that the occurrence of mycotoxins will likely increase in Europe due to the ongoing climate change.

Keywords: Anaerobic digestion, Biogas, Feedstock conservation, Fungal mycotoxins, Inhibition, process failure

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344 The Effects of Bisphosphonates on Osteonecrosis of Jaw Bone: A Stem Cell Perspective

Authors: Huseyin Apdik, Aysegul Dogan, Selami Demirci, Ezgi Avsar Apdik, Fikrettin Sahin

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Mesenchymal stem cells (MSCs) are crucial cell types for bone maintenance and growth along with resident bone progenitor cells providing bone tissue integrity during osteogenesis and skeletal growth. Any deficiency in this regulation would result in vital bone diseases. Of those, osteoporosis, characterized by a reduction in bone mass and mineral density, is a critical skeletal disease for especially elderly people. The commonly used drugs for the osteoporosis treatment are bisphosphonates (BPs). The most prominent role of BPs is to prevent bone resorption arisen from high osteoclast activity. However, administrations of bisphosphonates may also cause bisphosphonate-induced osteonecrosis of the jaw (BIONJ). Up to the present, the researchers have proposed several circumstances for BIONJ. However, effects of long-term and/or high dose usage of BPs on stem cell’s proliferation, survival, differentiation or maintenance capacity have not been evaluated yet. The present study will be held to; figure out BPs’ effects on MSCs in vitro in the aspect of cell proliferation and toxicity, migration, angiogenic activity, lineage specific gene and protein expression levels, mesenchymal stem cell properties and potential signaling pathways affected by BP treatment. Firstly, mesenchymal stem cell characteristics of Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs) were proved using flow cytometry analysis. Cell viability analysis was completed to determine the cytotoxic effects of BPs (Zoledronate (Zol), Alendronate (Ale) and Risedronate (Ris)) on DPSCs and PDLSCs by the 3-(4,5-di-methyl-thiazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfo-phenyl)-2H-tetrazolium (MTS) assay. Non-toxic concentrations of BPs were determined at 24 h under growth condition, and at 21 days under osteogenic differentiation condition for both cells. The scratch assay was performed to evaluate their migration capacity under the usage of determined of BPs concentrations at 24 h. The results revealed that while the scratch closure is 70% in the control group for DPSCs, it was 57%, 66% and 66% in Zol, Ale and Ris groups, respectively. For PDLSs, while wound closure is 71% in control group, it was 65%, 66% and 66% in Zol, Ale and Ris groups, respectively. As future experiments, tube formation assay and aortic ring assay will be done to determinate angiogenesis abilities of DPSCs and PDLSCs treated with BPs. Expression levels of osteogenic differentiation marker genes involved in bone development will be determined using real time-polymerase change reaction (RT-PCR) assay and expression profiles of important proteins involved in osteogenesis will be evaluated using western blotting assay for osteogenically differentiated MSCs treated with or without BPs. In addition to these, von Kossa staining will be performed to measure calcium mineralization status of MSCs.

Keywords: bisphosphonates, bisphosphonate-induced osteonecrosis of the jaw, mesenchymal stem cells, osteogenesis

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343 The Role of Establishing Zakat-Based Finance in Alleviating Poverty in the Muslim World

Authors: Khan Md. Abdus Subhan, Rabeya Bushra

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The management of Intellectual Property (IP) in museums can be complex and challenging, as it requires balancing access and control. On the one hand, museums must ensure that they have balanced permissions to display works in their collections and make them accessible to the public. On the other hand, they must also protect the rights of creators and owners of works and ensure that they are not infringing on IP rights. Intellectual property has become an increasingly important aspect of museum operations in the digital age. Museums hold a vast array of cultural assets in their collections, many of which have significant value as IP assets. The balanced management of IP in museums can help generate additional revenue and promote cultural heritage while also protecting the rights of the museum and its collections. Digital technologies have greatly impacted the way museums manage IP, providing new opportunities for revenue generation through e-commerce and licensing while also presenting new challenges related to IP protection and management. Museums must take a comprehensive approach to IP management, leveraging digital technologies, protecting IP rights, and engaging in licensing and e-commerce activities to maximize income and the economy of countries through the strong management of cultural institutions. Overall, the balanced management of IP in museums is crucial for ensuring the sustainability of museum operations and for preserving cultural heritage for future generations. By taking a balanced approach to identifying museum IP assets, museums can generate revenues and secure their financial sustainability to ensure the long-term preservation of their cultural heritage. We can divide IP assets in museums into two kinds: collection IP and museum-generated IP. Certain museums become confused and lose sight of their mission when trying to leverage collections-based IP. This was the case at the German State Museum in Berlin when the museum made 100 replicas from the Nefertiti bust and wrote under the replicas all rights reserved to the Berlin Museum and issued a certificate to prevent any person or Institution from reproducing any replica from this bust. The implications of IP in museums are far-reaching and can have significant impacts on the preservation of cultural heritage, the dissemination of information, and the development of educational programs. As such, it is important for museums to have a comprehensive understanding of IP laws and regulations and to properly manage IP to avoid legal liability, damage to reputation, and loss of revenue. The research aims to highlight the importance and role of intellectual property in museums and provide some illustrative examples of this.

Keywords: zakat, economic development, Muslim world, poverty alleviation.

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342 Histogenesis of the Stomach of Pre-Hatching Quail: A Light and Electron Microscopic Study

Authors: Soha A Soliman, Yasser A Ahmed, Mohamed A Khalaf

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Although the enormous literature describing the histology of the stomach of different avian species during the posthatching development, the available literature on the pre-hatching development of quail stomach development is scanty. Thus, the current study was undertaken to provide a careful description of the main histological events during the embryonic development of quail stomach. To achieve this aim, daily histological specimens from the stomach of quail of 4 days post-incubation till the day 17 (few hours before hatching) were examined with light microscopy. The current study showed that the primitive gut tube of the embryonic quail appeared at the 4th day post incubation, and both parts of stomach (proventriculus and gizzard) were similar in structure and composed of endodermal epithelium of pseudostratified type surrounded by undifferentiated mesenchymal tissue. The sequences of the developmental events in the gut tube were preceded in a cranio-caudal pattern. By the 5th day, the endodermal covering of the primitive proventriculus gave rise to sac-like invaginations. The primitive gizzard was distinguished into thick-walled bodies and thin-walled sacs. In the 6th day, the prospective proventricular glandular epithelium became canalized and the muscular layer was developed in the cranial part of the proventriculus, whereas the primitive muscular coat of the gizzard was represented by a layer of condensed mesenchyme. In the 7th day, the proventricular glandular epithelial invaginations increased in depth and number, while, the muscularis mucosa and the muscular layer began to be distinguished. In the 8th day, the myoblasts differentiated into spindle shaped smooth muscle fibers. In the 10th day, branching of the proventricular glands began. The branching continued later on. The surface and the glandular epithelium were transformed into simple columnar type in the 12th day. The epithelial covering of the gizzard gave rise to tubular invaginations lined by simple cuboidal epithelium and the surface epithelium became simple columnar. Canalization of the tubular glands was recognized in the 14th day. In the 15th day, the proventricular surface epithelium invaginated in an concentric manner around a central cavity to form immature secretory units. The central cavity was lined by eosinophilic cells which form the ductal epithelia. The peripheral lamellae were lined by basophilic cells; the undifferentiated oxyntico-peptic cells. Entero-endocrine cells stained positive for silver impregnation in the proventricular glands. The mucosal folding in the gizzard appeared in the 15th day to form the plicae and the sulci. The wall of the proventriculus and gizzard in the 17th day acquired the main histological features of post-hatching birds, but neither the surface nor the ductal epithelium were differentiated to mucous producing cells. The current results shoed be considered in the molecular developmental studies.

Keywords: quail, proventriculus, gizzard, pre-hatching, histology

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341 Bio-Functionalized Silk Nanofibers for Peripheral Nerve Regeneration

Authors: Kayla Belanger, Pascale Vigneron, Guy Schlatter, Bernard Devauchelle, Christophe Egles

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A severe injury to a peripheral nerve leads to its degeneration and the loss of sensory and motor function. To this day, there still lacks a more effective alternative to the autograft which has long been considered the gold standard for nerve repair. In order to overcome the numerous drawbacks of the autograft, tissue engineered biomaterials may be effective alternatives. Silk fibroin is a favorable biomaterial due to its many advantageous properties such as its biocompatibility, its biodegradability, and its robust mechanical properties. In this study, bio-mimicking multi-channeled nerve guidance conduits made of aligned nanofibers achieved by electrospinning were functionalized with signaling biomolecules and were tested in vitro and in vivo for nerve regeneration support. Silk fibroin (SF) extracted directly from silkworm cocoons was put in solution at a concentration of 10wt%. Poly(ethylene oxide) (PEO) was added to the resulting SF solution to increase solution viscosity and the following three electrospinning solutions were made: (1) SF/PEO solution, (2) SF/PEO solution with nerve growth factor and ciliary neurotrophic factor, and (3) SF/PEO solution with nerve growth factor and neurotrophin-3. Each of these solutions was electrospun into a multi-layer architecture to obtain mechanically optimized aligned nanofibrous mats. For in vitro studies, aligned fibers were treated to induce β-sheet formation and thoroughly rinsed to eliminate presence of PEO. Each material was tested using rat embryo neuron cultures to evaluate neurite extension and the interaction with bio-functionalized or non-functionalized aligned fibers. For in vivo studies, the mats were rolled into 5mm long multi-, micro-channeled conduits then treated and thoroughly rinsed. The conduits were each subsequently implanted between a severed rat sciatic nerve. The effectiveness of nerve repair over a period of 8 months was extensively evaluated by cross-referencing electrophysiological, histological, and movement analysis results to comprehensively evaluate the progression of nerve repair. In vitro results show a more favorable interaction between growing neurons and bio-functionalized silk fibers compared to pure silk fibers. Neurites can also be seen having extended unidirectionally along the alignment of the nanofibers which confirms a guidance factor for the electrospun material. The in vivo study has produced positive results for the regeneration of the sciatic nerve over the length of the study, showing contrasts between the bio-functionalized material and the non-functionalized material along with comparisons to the experimental control. Nerve regeneration has been evaluated not only by histological analysis, but also by electrophysiological assessment and motion analysis of two separate natural movements. By studying these three components in parallel, the most comprehensive evaluation of nerve repair for the conduit designs can be made which can, therefore, more accurately depict their overall effectiveness. This work was supported by La Région Picardie and FEDER.

Keywords: electrospinning, nerve guidance conduit, peripheral nerve regeneration, silk fibroin

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340 Effects of Oxidized LDL in M2 Macrophages: Implications in Atherosclerosis

Authors: Fernanda Gonçalves, Karla Alcântara, Vanessa Moura, Patrícia Nolasco, Jorge Kalil, Maristela Hernandez

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Introduction: Atherosclerosis is a chronic disease where two striking features are observed: retention of lipids and inflammation. Understanding the interaction between immune cells and lipoproteins involved in atherogenesis are urgent challenges, since cardiovascular diseases are the leading cause of death worldwide. Macrophages are critical to the development of atherosclerotic plaques and in the perpetuation of inflammation in these lesions. These cells are also directly involved in unstable plaque rupture. Recently different populations of macrophages are being identified in atherosclerotic lesions. Although the presence of M2 macrophages (macrophages activated by the alternative pathway, eg. The IL-4) has been identified, the function of these cells in atherosclerosis is not yet defined. M2 macrophages have a high endocytic capacity, they promote remodeling of tissues and to have anti-inflammatory activity. However, in atherosclerosis, especially unstable plaques, severe inflammatory reaction, accumulation of cellular debris and intense degradation of the tissue is observed. Thus, it is possible that the M2 macrophages have altered function (phenotype) in atherosclerosis. Objective: Our aim is to evaluate if the presence of oxidized LDL alters the phenotype and function of M2 macrophages in vitro. Methods: For this, we will evaluate whether the addition of lipoprotein in M2 macrophages differentiated in vitro with IL -4 induces 1) a reduction in the secretion of anti-inflammatory cytokines (CBA and ELISA), 2) secretion of inflammatory cytokines (CBA and ELISA), 3) expression of cell activation markers (Flow cytometry), 4) alteration in gene expression of molecules adhesion and extracellular matrix (Real-Time PCR) and 5) Matrix degradation (confocal microscopy). Results: In oxLDL stimulated M2 macrophages cultures we did not find any differences in the expression of the cell surface markers tested, including: HLA-DR, CD80, CD86, CD206, CD163 and CD36. Also, cultures stimulated with oxLDL had similar phagocytic capacity when compared to unstimulated cells. However, in the supernatant of these cultures an increase in the secretion of the pro-inflammatory cytokine IL-8 was detected. No significant changes where observed in IL-6, IL-10, IL-12 and IL-1b levels. The culture supernatant also induced massive extracellular matrix (produced by mouse embryo fibroblast) filaments degradation. When evaluating the expression of 84 extracellular matrix and adhesion molecules genes, we observed that the stimulation of oxLDL in M2 macrophages decreased 47% of the genes and increased the expression of only 3% of the genes. In particular we noted that oxLDL inhibit the expression of 60% of the genes constituents of extracellular matrix and collagen expressed by these cells, including fibronectin1 and collagen VI. We also observed a decrease in the expression of matrix protease inhibitors, such as TIMP 2. On the opposite, the matricellular protein thrombospondin had a 12 fold increase in gene expression. In the presence of native LDL 90% of the genes had no altered expression. Conclusion: M2 macrophages stimulated with oxLDL secrete the pro-inflammatory cytokine IL-8, have an altered extracellular matrix constituents gene expression, and promote the degradation of extracellular matrix. M2 macrophages may contribute to the perpetuation of inflammation in atherosclerosis and to plaque rupture.

Keywords: atherosclerosis, LDL, macrophages, m2

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339 Examining the Role of Farmer-Centered Participatory Action Learning in Building Sustainable Communities in Rural Haiti

Authors: Charles St. Geste, Michael Neumann, Catherine Twohig

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Our primary aim is to examine farmer-centered participatory action learning as a tool to improve agricultural production, build resilience to climate shocks and, more broadly, advance community-driven solutions for sustainable development in rural communities across Haiti. For over six years, sixty plus farmers from Deslandes, Haiti, organized in three traditional work groups called konbits, have designed and tested low-input agroecology techniques as part of the Konbit Vanyan Kapab Pwoje Agroekoloji. The project utilizes a participatory action learning approach, emphasizing social inclusion, building on local knowledge, experiential learning, active farmer participation in trial design and evaluation, and cross-community sharing. Mixed methods were used to evaluate changes in knowledge and adoption of agroecology techniques, confidence in advancing agroecology locally, and innovation among Konbit Vanyan Kapab farmers. While skill and knowledge in application of agroecology techniques varied among individual farmers, a majority of farmers successfully adopted techniques outside of the trial farms. The use of agroecology techniques on trial and individual farms has doubled crop production in many cases. Farm income has also increased, and farmers report less damage to crops and property caused by extreme weather events. Furthermore, participatory action strategies have led to greater local self-determination and greater capacity for sustainable community development. With increased self-confidence and the knowledge and skills acquired from participating in the project, farmers prioritized sharing their successful techniques with other farmers and have developed a farmer-to-farmer training program that incorporates participatory action learning. Using adult education methods, farmers, trained as agroecology educators, are currently providing training in sustainable farming practices to farmers from five villages in three departments across Haiti. Konbit Vanyan Kapab farmers have also begun testing production of value-added food products, including a dried soup mix and tea. Key factors for success include: opportunities for farmers to actively participate in all phases of the project, group diversity, resources for application of agroecology techniques, focus on group processes and overcoming local barriers to inclusive decision-making.

Keywords: agroecology, participatory action learning, rural Haiti, sustainable community development

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338 Response Analysis of a Steel Reinforced Concrete High-Rise Building during the 2011 Tohoku Earthquake

Authors: Naohiro Nakamura, Takuya Kinoshita, Hiroshi Fukuyama

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The 2011 off The Pacific Coast of Tohoku Earthquake caused considerable damage to wide areas of eastern Japan. A large number of earthquake observation records were obtained at various places. To design more earthquake-resistant buildings and improve earthquake disaster prevention, it is necessary to utilize these data to analyze and evaluate the behavior of a building during an earthquake. This paper presents an earthquake response simulation analysis (hereafter a seismic response analysis) that was conducted using data recorded during the main earthquake (hereafter the main shock) as well as the earthquakes before and after it. The data were obtained at a high-rise steel-reinforced concrete (SRC) building in the bay area of Tokyo. We first give an overview of the building, along with the characteristics of the earthquake motion and the building during the main shock. The data indicate that there was a change in the natural period before and after the earthquake. Next, we present the results of our seismic response analysis. First, the analysis model and conditions are shown, and then, the analysis result is compared with the observational records. Using the analysis result, we then study the effect of soil-structure interaction on the response of the building. By identifying the characteristics of the building during the earthquake (i.e., the 1st natural period and the 1st damping ratio) by the Auto-Regressive eXogenous (ARX) model, we compare the analysis result with the observational records so as to evaluate the accuracy of the response analysis. In this study, a lumped-mass system SR model was used to conduct a seismic response analysis using observational data as input waves. The main results of this study are as follows: 1) The observational records of the 3/11 main shock put it between a level 1 and level 2 earthquake. The result of the ground response analysis showed that the maximum shear strain in the ground was about 0.1% and that the possibility of liquefaction occurring was low. 2) During the 3/11 main shock, the observed wave showed that the eigenperiod of the building became longer; this behavior could be generally reproduced in the response analysis. This prolonged eigenperiod was due to the nonlinearity of the superstructure, and the effect of the nonlinearity of the ground seems to have been small. 3) As for the 4/11 aftershock, a continuous analysis in which the subject seismic wave was input after the 3/11 main shock was input was conducted. The analyzed values generally corresponded well with the observed values. This means that the effect of the nonlinearity of the main shock was retained by the building. It is important to consider this when conducting the response evaluation. 4) The first period and the damping ratio during a vibration were evaluated by an ARX model. Our results show that the response analysis model in this study is generally good at estimating a change in the response of the building during a vibration.

Keywords: ARX model, response analysis, SRC building, the 2011 off the Pacific Coast of Tohoku Earthquake

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337 Computational and Experimental Study of the Mechanics of Heart Tube Formation in the Chick Embryo

Authors: Hadi S. Hosseini, Larry A. Taber

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In the embryo, heart is initially a simple tubular structure that undergoes complex morphological changes as it transforms into a four-chambered pump. This work focuses on mechanisms that create heart tube (HT). The early embryo is composed of three relatively flat primary germ layers called endoderm, mesoderm, and ectoderm. Precardiac cells located within bilateral regions of the mesoderm called heart fields (HFs) fold and fuse along the embryonic midline to create the HT. The right and left halves of this plate fold symmetrically to bring their upper edges into contact along the midline, where they fuse. In a region near the fusion line, these layers then separate to generate the primitive HT and foregut, which then extend vertically. The anterior intestinal portal (AIP) is the opening at the caudal end of the foregut, which descends as the HT lengthens. The biomechanical mechanisms that drive this folding are poorly understood. Our central hypothesis is that folding is caused by differences in growth between the endoderm and mesoderm while subsequent extension is driven by contraction along the AIP. The feasibility of this hypothesis is examined using experiments with chick embryos and finite-element modeling (FEM). Fertilized white Leghorn chicken eggs were incubated for approximately 22-33 hours until appropriate Hamburger and Hamilton stage (HH5 to HH9) was reached. To inhibit contraction, embryos were cultured in media containing blebbistatin (myosin II inhibitor) for 18h. Three-dimensional models were created using ABAQUS (D. S. Simulia). The initial geometry consists of a flat plate including two layers representing the mesoderm and endoderm. Tissue was considered as a nonlinear elastic material with growth and contraction (negative growth) simulated using a theory, in which the total deformation gradient is given by F=F^*.G, where G is growth tensor and F* is the elastic deformation gradient tensor. In embryos exposed to blebbistatin, initial folding and AIP descension occurred normally. However, after HFs partially fused to create the upper part of the HT, fusion, and AIP descension stopped, and the HT failed to grow longer. These results suggest that cytoskeletal contraction is required only for the later stages of HT formation. In the model, a larger biaxial growth rate in the mesoderm compared to the endoderm causes the bilayered plate to bend ventrally, as the upper edge moves toward the midline, where it 'fuses' with the other half . This folding creates the upper section of the HT, as well as the foregut pocket bordered by the AIP. After this phase completes by stage HH7, contraction along the arch-shaped AIP pulls the lower edge of the plate downward, stretching the two layers. Results given by model are in reasonable agreement with experimental data for the shape of HT, as well as patterns of stress and strain. In conclusion, results of our study support our hypothesis for the creation of the heart tube.

Keywords: heart tube formation, FEM, chick embryo, biomechanics

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336 Gold Nanoprobes Assay for the Identification of Foodborn Pathogens Such as Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

Abstract:

Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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335 Mechanical and Material Characterization on the High Nitrogen Supersaturated Tool Steels for Die-Technology

Authors: Tatsuhiko Aizawa, Hiroshi Morita

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The tool steels such as SKD11 and SKH51 have been utilized as punch and die substrates for cold stamping, forging, and fine blanking processes. The heat-treated SKD11 punches with the hardness of 700 HV wrought well in the stamping of SPCC, normal steel plates, and non-ferrous alloy such as a brass sheet. However, they suffered from severe damage in the fine blanking process of smaller holes than 1.5 mm in diameter. Under the high aspect ratio of punch length to diameter, an elastoplastic bucking of slender punches occurred on the production line. The heat-treated punches had a risk of chipping at their edges. To be free from those damages, the blanking punch must have sufficient rigidity and strength at the same time. In the present paper, the small-hole blanking punch with a dual toughness structure was proposed to provide a solution to this engineering issue in production. The low-temperature plasma nitriding process was utilized to form the nitrogen supersaturated thick layer into the original SKD11 punch. Through the plasma nitriding at 673 K for 14.4 ks, the nitrogen supersaturated layer, with the thickness of 50 μm and without nitride precipitates, was formed as a high nitrogen steel (HNS) layer surrounding the original SKD11 punch. In this two-zone structured SKD11 punch, the surface hardness increased from 700 HV for the heat-treated SKD11 to 1400 HV. This outer high nitrogen SKD11 (HN-SKD11) layer had a homogeneous nitrogen solute depth profile with a nitrogen solute content plateau of 4 mass% till the border between the outer HN-SKD11 layer and the original SKD11 matrix. When stamping the brass sheet with the thickness of 1 mm by using this dually toughened SKD11 punch, the punch life was extended from 500 K shots to 10000 K shots to attain a much more stable production line to yield the brass American snaps. Furthermore, with the aid of the masking technique, the punch side surface layer with the thickness of 50 μm was modified by this high nitrogen super-saturation process to have a stripe structure where the un-nitrided SKD11 and the HN-SKD11 layers were alternatively aligned from the punch head to the punch bottom. This flexible structuring promoted the mechanical integrity of total rigidity and toughness as a punch with an extremely small diameter.

Keywords: high nitrogen supersaturation, semi-dry cold stamping, solid solution hardening, tool steel dies, low temperature nitriding, dual toughness structure, extremely small diameter punch

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334 Plantar Neuro-Receptor Activation in Total Knee Arthroplasty Patients: Impact on Clinical Function, Pain, and Stiffness - A Randomized Controlled Trial

Authors: Woolfrey K., Woolfrey M., Bolton C. L., Warchuk D.

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Objectives: Osteoarthritis is the most common joint disease of adults worldwide. Despite total knee arthroplasty (TKA) demonstrating high levels of success, 20% of patients report dissatisfaction with their result. VOXX Wellness Stasis Socks are embedded with a proprietary pattern of neuro-receptor activation points that have been proven to activate a precise neuro-response, according to the pattern theory of haptic perception, which stimulates improvements in pain and function. The use of this technology in TKA patients may prove beneficial as an adjunct to recovery as many patients suffer from deficits to their proprioceptive system caused by ligamentous damage and alterations to mechanoreceptors during the procedure. We hypothesized that VOXX Wellness Stasis Socks are a safe, cost-effective, and easily scalable strategy to support TKA patients through their recovery. Design: Double-blinded, placebo-controlled randomized trial. Participants: Patients scheduled to receive TKA were considered eligible for inclusion in the trial. Interventions: Intervention group (I): VOXX Wellness Stasis socks containing receptor point-activation technology. Control group (C): VOXX Wellness Stasis socks without receptor point-activation technology. Sock use during the waking hours x 6 weeks. Main Outcome Measures: Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) questionnaire completed at baseline, 2 weeks, and 6 weeks to assess pain, stiffness, and physical function. Results: Data analysis using SPSS software. P-values, effect sizes, and confidence intervals are reported to assess clinical relevance of the finding. Physical status classifications were compared using t-test. Within-subject and between-subject differences in the mean WOMAC were analyzed by ANOVA. Effect size was analyzed using Cramer’s V. Consistent improvement in WOMAC scores for pain and stiffness at 2 weeks post op in the I over the C group. The womac scores assessing physical function showed a consistent improvement at both 2 and 6 weeks post op in the I group compared to C group. Conclusions: VOXX proved to be a low cost, safe intervention in TKA to help patients improve with regard to pain, stiffness, and physical function. Disclosures: None

Keywords: osteoarthritis, RCT, pain management, total knee arthroplasty

Procedia PDF Downloads 528
333 Rural Farmers-Herdsmen Conflicts, State Mediation Failure and Prospects of Traditional Institutions’ Intervention in Southwest Nigeria

Authors: Grace Adebo

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Rural Farmers-herdsmen conflicts have resulted in a large number of causalities in many parts of Nigeria. Herds of cattle have died, while farmers recorded inestimable losses of their crops and harvests. The overall consequences have impacted negatively on food security across the country. There are divided opinions by scholars, agricultural experts and conflict analysts on the root causes of the conflicts and why traditional institutional interventions are ineffective in resolving the crisis. The study, therefore, aims to investigate the fundamentality of the conflicts’ causes in Southwest Nigeria and the correlates between traditional institutional authorities’ intervention and farmers-herdsmen conflicts in Southwest Nigeria. A structured interview schedule and focus group discussion were employed to elicit information from 180 farmers and 48 herdsmen selected through a multistage sampling procedure from the conflict zones in Southwest Nigeria. Collected data were analyzed using frequency counts, percentages, means and the Relative Importance Index (RII). The study found that climate change effects, farmland encroachment, crop damage, theft, and competition for land and water resources and pollution were the root causes of the violent herders-rural farmer’s clashes. The quest for wealth acquisition by some traditional rulers and some notable individuals in the conflict neighborhoods, occasioned tribal-mix herds possession and, thus undermining local institutional interventions and perverting justice through weak conflict resolution strategies, therefore, fueling further conflicts. Most farmers in the conflict zones have abandoned their farms for fear of death. This coupled with physical, social, economic and psychological consequences have deepened food insecurity and impaired the economic conditions of the herdsmen and the farmers. Currently, there are no mutually established mediation mechanisms as most states are opposed to the enactment of grazing laws to protect territorial encroachments of lands and subsequent multiplication of the herdsmen. It is suggested that government and Non-Governmental Organisation (NGOs) should encourage a functional stakeholder's forum for sustainable conflict resolution and establish a compensation scheme for losses incurred while extension agents are equipped with knowledge on conflict management strategies for peace attainment with the envisioned goal of achieving sustainable livelihoods and food security in Southwest Nigeria.

Keywords: conflict resolution, food security, herdsmen-farmers conflict, sustainable livelihoods, traditional institutions

Procedia PDF Downloads 108
332 Osteosuture in Fixation of Displaced Lateral Third Clavicle Fractures: A Case Report

Authors: Patrícia Pires, Renata Vaz, Bárbara Teles, Marco Pato, Pedro Beckert

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Introduction: The management of lateral third clavicle fractures can be challenging due to difficulty in distinguishing subtle variations in the fracture pattern, which may be suggestive of potential fracture instability. They occur most often in men between 30 and 50 years of age, and in individuals over 70 years of age, its distribution is equal between both men and women. These fractures account for 10%–30% of all clavicle fractures and roughly 30%–45% of all clavicle nonunion fractures. Lateral third clavicle fractures may be treated conservatively or surgically, and there is no gold standard, although the risk of nonunion or pseudoarthrosis impacts the recommendation of surgical treatment when these fractures are unstable. There are many strategies for surgical treatment, including locking plates, hook plates fixation, coracoclavicular fixation using suture anchors, devices or screws, tension band fixation with suture or wire, transacromial Kirschner wire fixation and arthroscopically assisted techniques. Whenever taking the hardware into consideration, we must not disregard that obtaining adequate lateral fixation of small fragments is a difficult task, and plates are more associated to local irritation. The aim of the appropriate treatment is to ensure fracture healing and a rapid return to preinjury activities of daily living but, as explained, definitive treatment strategies have not been established and the variety of techniques avalilable add up to the discussion of this topic. Methods and Results: We present a clinical case of a 43-year-old man with the diagnosis of a lateral third clavicle fracture (Neer IIC) in the sequence of a fall on his right shoulder after a bicycle fall. He was operated three days after the injury, and through K-wire temporary fixation and indirect reduction using a ZipTight, he underwent osteosynthesis with an interfragmentary figure-of-eight tension band with polydioxanone suture (PDS). Two weeks later, there was a good aligment. He kept the sling until 6 weeks pos-op, avoiding efforts. At 7-weeks pos-op, there was still a good aligment, starting the physiotherapy exercises. After 10 months, he had no limitation in mobility or pain and returned to work with complete recovery in strength. Conclusion: Some distal clavicle fractures may be conservatively treated, but it is widely accepted that unstable fractures require surgical treatment to obtain superior clinical outcomes. In the clinical case presented, the authors chose an osteosuture technique due to the fracture pattern, its location. Since there isn´t a consensus on the prefered fixation method, it is important for surgeons to be skilled in various techniques and decide with their patient which approach is most appropriate for them, weighting the risk-benefit of each method. For instance, with the suture technique used, there is no wire migration or breakage, and it doesn´t require a reoperation for hardware removal; there is also less tissue exposure since it requires a smaller approach in comparison to the plate fixation and avoids cuff tears like the hook plate. The good clinical outcome on this case report serves the purpose of expanding the consideration of this method has a therapeutic option.

Keywords: lateral third, clavicle, suture, fixation

Procedia PDF Downloads 74
331 The Influences of Facies and Fine Kaolinite Formation Migration on Sandstones’ Reservoir Quality, Sarir Formation, Sirt Basin Libya

Authors: Faraj M. Elkhatri, Hana Ali Alafi

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The spatial and temporal distribution of diagenetic alterations related impact on the reservoir quality of the Sarir Formation. (present-day burial depth of about 9000 feet) Depositional facies and diagenetic alterations are the main controls on reservoir quality of Sarir Formation Sirt Basin Libya; these based on lithology and grain size as well as authigenic clay mineral types and their distributions. However, petrology investigation obtained on study area with five sandstone wells concentrated on main rock components and the parameters that may have impacts on reservoirs. the main authigenic clay minerals are kaolinite and dickite, these investigations have confirmed by X.R.D analysis and clay fraction. mainly Kaolinite and Dickite were extensively presented on all of wells with high amounts. As well as trace of detrital smectite and less amounts of illitized mud-matrix are possibly found by SEM image. Thin layers of clay presented as clay-grain coatings in local depth interpreted as remains of dissolved clay matrix is partly transformed into kaolinite adjacent and towards pore throat. This also may have impacts on most of the pore throats of this sandstone which are open and relatively clean with some of fine martial have been formed on occluded pores. This material is identified by EDS analysis to be collections of not only kaolinite booklets but also small disaggregated kaolinite platelets derived from the disaggregation of larger kaolinite booklets. These patches of kaolinite not only fill this pore, but also coat some of the surrounding framework grains. Quartz grains often enlarged by authigenic quartz overgrowths partially occlude and reduce porosity. Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM) was conducted on the post-test samples to examine any mud filtrate particles that may be in the pore throats. Semi-qualitative elemental data on selected minerals observed during the SEM study were obtained through the use of an Energy Dispersive Spectroscopy (EDS) unit. The samples showed mostly clean open pore throats, with limited occlusion by kaolinite. very fine-grained elemental combinations (Si/Al/Na/Cl, Si/Al Ca/Cl/Ti, and Qtz/Ti) have been identified and conformed by EDS analysis. However, the identification of the fine grained disaggregated material as mainly kaolinite though study area.

Keywords: fine migration, formation damage, kaolinite, soled bulging.

Procedia PDF Downloads 66
330 Generative Behaviors and Psychological Well-Being in Mexican Elders

Authors: Ana L. Gonzalez-Celis, Edgardo Ruiz-Carrillo, Karina Reyes-Jarquin, Margarita Chavez-Becerra

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Since recent decades, the aging has been viewed from a more positive perspective, where is not only about losses and damage, but also about being on a stage where you can enjoy life and live with well-being and quality of life. The challenge to feel better is to find those resources that seniors have. For that reason, psychological well-being has shown interest in the study of the affect and life satisfaction (hedonic well-being), while from a more recent tradition, focus on the development of capabilities and the personal growth, considering both as the main indicators of the quality of life. A resource that can be used in the later age is generativity, which refers to the ability of older people to develop and grow through activities that contribute with the improvement of the context in which they live and participate. In this way the generative interest is understood as a favourable attitude that contribute to the common benefit while strengthening and enriching the social institutions, to ensure continuity between generations and social development. On the other hand, generative behavior, differentiating from generative interest, is the expression of that attitude reflected in activities that make a social contribution and a benefit for generations to come. Hence the purpose of the research was to test if there is an association between the generative behaviour type and the psychological well-being with their dimensions. For this reason 188 Mexican adults from 60 to 94 years old (M = 69.78), 67% women, 33% men, completed two instruments: The Ryff’s Well-Being Scales to measure psychological well-being with 39 items with two dimensions (Hedonic and Eudaimonic well-being), and the Loyola’s Generative Behaviors Scale, grouped in five categories: Knowledge transmitted to the next generation, things to be remember, creativity, be productive, contribution to the community, and responsibility of other people. In addition, the socio-demographic data sheet was tested, and self-reported health status. The results indicated that the psychological well-being and its dimensions were significantly associated with the presence of generative behavior, where the level of well-being was higher when the frequency of some generative behaviour excelled; finding that the behavior with greater psychological well-being (M = 81.04, SD = 8.18) was "things to be remembered"; while with greater hedonic well-being (M = 73.39, SD = 12.19) was the behavior "responsibility of other people"; and with greater Eudaimonic well-being (M = 84.61, SD = 6.63), was the behavior "things to be remembered”. The most important findings highlight the importance of generative behaviors in adulthood, finding empirical evidence that the generativity in the last stage of life is associated with well-being. However, by finding differences in the types of generative behaviors at the level of well-being, is proposed the idea that generativity is not situated as an isolated construct, but needs other contextualized and related constructs that can simultaneously operate at different levels, taking into account the relationship between the environment and the individual, encompassing both the social and psychological dimension.

Keywords: eudaimonic well-being, generativity, hedonic well-being, Mexican elders, psychological well-being

Procedia PDF Downloads 266
329 Previously Undescribed Cardiac Abnormalities in Two Unrelated Autistic Males with Causative Variants in CHD8

Authors: Mariia A. Parfenenko, Ilya S. Dantsev, Sergei V. Bochenkov, Natalia V. Vinogradova, Olga S. Groznova, Victoria Yu. Voinova

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Introduction: Autism is the most common neurodevelopmental disorder. Autism is characterized by difficulties in social interaction and adherence to stereotypic behavioral patterns and frequently co-occurs with epilepsy, intellectual disabilities, connective tissue disorders, and other conditions. CHD8 codes for chromodomain-helicase-DNA-binding protein 8 - a chromatin remodeler that regulates cellular proliferation and neurodevelopment in embryogenesis. CHD8 is one of the genes most frequently involved in autism. Patients and methods: 2 unrelated male patients, P3 and P12, aged 3 and 12 years old, underwent whole genome sequencing, which determined that they both had different likely pathogenic variants, both previously undescribed in literature. Sanger sequencing later determined that P12 inherited the variant from his affected mother. Results: P3 and P12 presented with autism, a developmental delay, ataxia, sleep disorders, overgrowth, and macrocephaly, as well as other clinical features typically present in patients with causative variants in CHD8. The mother of P12 also has autistic traits, as well as ataxia, hypotonia, sleep disorders, and other symptoms. However, P3 and P12 also have different cardiac abnormalities. P3 had signs of a repolarization disorder: a flattened T wave in the III and aVF derivations and a negative T wave in the V1-V2 derivations. He also had structural valve anomalies with associated regurgitation, local contractility impairment of the left ventricular, and diastolic dysfunction of the right ventricle. Meanwhile, P12 had Wolff-Parkinson-White syndrome and underwent radiofrequency ablation at the age of 2 years. At the time of observation, P12 had mild sinus arrhythmia and an incomplete right bundle branch block, as well as arterial hypertension. Discussion: Cardiac abnormalities were not previously reported in patients with causative variants in CHD8. The underlying mechanism for the formation of those abnormalities is currently unknown. However, the two hypotheses are either a disordered interaction with CHD7 – another chromodomain remodeler known to be directly involved in the cardiophenotype of CHARGE syndrome – a rare condition characterized by coloboma, heart defects and growth abnormalities, or the disrupted functioning of CHD8 as an A-Kinase Anchoring Protein, which are known to modulate cardiac function. Conclusion: We observed 2 unrelated autistic males with likely pathogenic variants in CHD8 that presented with typical symptoms of CHD8-related neurodevelopmental disorder, as well as cardiac abnormalities. Cardiac abnormalities have, until now, been considered uncharacteristic for patients with causative variants in CHD8. Further accumulation of data, including experimental evidence of the involvement of CHD8 in heart formation, will elucidate the mechanism underlying the cardiophenotype of those patients. Acknowledgements: Molecular genetic testing of the patients was made possible by the Charity Fund for medical and social genetic aid projects «Life Genome.»

Keywords: autism spectrum disorders, chromodomain-helicase-DNA-binding protein 8, neurodevelopmental disorder, cardio phenotype

Procedia PDF Downloads 84
328 The Environmental Impact Assessment of Land Use Planning (Case Study: Tannery Industry in Al-Garma District)

Authors: Husam Abdulmuttaleb Hashim

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The environmental pollution problems represent a great challenge to the world, threatening to destroy all the evolution that mankind has reached, the organizations and associations that cares about environment are trying to warn the world from the forthcoming danger resulted from excessive use of nature resources and consuming it without looking to the damage happened as a result of unfair use of it. Most of the urban centers suffers from the environmental pollution problems and health, economic, and social dangers resulted from this pollution, and while the land use planning is responsible for distributing different uses in urban centers and controlling the interactions between these uses to reach a homogeneous and perfect state for the different activities in cities, the occurrence of environmental problems in the shade of existing land use planning operation refers to the disorder or insufficiency in this operation which leads to presence of such problems, and this disorder lays in lack of sufficient importance to the environmental considerations during the land use planning operations and setting up the master plan, so the research start to study this problem and finding solutions for it, the research assumes that using accurate and scientific methods in early stages of land use planning operation will prevent occurring of environmental pollution problems in the future, the research aims to study and show the importance of the environmental impact assessment method (EIA) as an important planning tool to investigate and predict the pollution ranges of the land use that has a polluting pattern in land use planning operation. This research encompasses the concept of environmental assessment and its kinds and clarifies environmental impact assessment and its contents, the research also dealt with urban planning concept and land use planning, it also dealt with the current situation of the case study (Al-Garma district) and the land use planning in it and explain the most polluting use on the environment which is the industrial land use represented in the tannery industries and then there was a stating of current situation of this land use and explaining its contents and environmental impacts resulted from it, and then we analyzed the tests applied by the researcher for water and soil, and perform environmental evaluation through applying environmental impact assessment matrix using the direct method to reveal the pollution ranges on the ambient environment of industrial land use, and we also applied the environmental and site limits and standards by using (GIS) and (AUTOCAD) to select the site of the best alternative of the industrial region in Al-Garma district after the research approved the unsuitability of its current site location for the environmental and site limitations, the research conducted some conclusions and recommendations regard clarifying the concluded facts and to set the proper solutions.

Keywords: EIA, pollution, tannery industry, land use planning

Procedia PDF Downloads 447
327 The Effect of Chloride Dioxide and High Concentration of CO2 Gas Injection on the Quality and Shelf-Life for Exporting Strawberry 'Maehyang' in Modified Atmosphere Condition

Authors: Hyuk Sung Yoon, In-Lee Choi, Mohammad Zahirul Islam, Jun Pill Baek, Ho-Min Kang

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The strawberry ‘Maehyang’ cultivated in South Korea has been increased to export to Southeast Asia. The degradation of quality often occurs in strawberries during short export period. Botrytis cinerea has been known to cause major damage to the export strawberries and the disease was caused during shipping and distribution. This study was conducted to find out the sterilized effect of chlorine dioxide(ClO2) gas and high concentration of CO2 gas injection for ‘Maehyang’ strawberry and it was packaged with oxygen transmission rate (OTR) films. The strawberry was harvested at 80% color changed stage and packaged with OTR film and perforated film (control). The treatments were a MAP used by with 20,000 cc·m-2·day·atm OTR film and gas injection in packages. The gas type of ClO2 and CO2 were injected into OTR film packages, and treatments were 6 mg/L ClO2, 15% CO2, and they were combined. The treated strawberries were stored at 3℃ for 30 days. Fresh weight loss rate was less than 1% in all OTR film packages but it was more than 15% in a perforated film treatment that showed severe deterioration of visual quality during storage. Carbon dioxide concentration within a package showed approximately 15% of the maximum CO2 concentration in all treatments except control until the 21st day, it was the tolerated range of maximum CO2 concentration of strawberry in recommended CA or MA conditions. But, it increased to almost 50% on the 30th day. Oxygen concentration showed a decrease down to approximately 0% in all treatments except control for 25 days. Ethylene concentration was shown to be steady until the 17th day, but it quickly increased on the 17th day and dropped down on the final storage day (30th day). All treatments did not show any significant differences in gas treatments. Firmness increased in CO2 (15%) and ClO2 (6mg/L) + CO2 (15%) treatments during storage. It might be the effect of high concentration CO2 known by reducing decay and cell wall degradation. The soluble solid decreased in all treatments during storage. These results were caused to use up the sugar by the increase of respiration during storage. The titratable acidity showed a similarity in all treatments. Incidence of fungi was 0% in CO2 (15%) and ClO2 (6mg/L)+ CO2 (15%), but was more than 20% in a perforated film treatment. Consequently, The result indicates that Chloride Dioxide(ClO2) and high concentration of CO2 inhibited fungi growth. Due to the fact that fresh weight loss rate and incidence of fungi were lower, the ClO2(6mg/L)+ CO2(15%) prove to be most efficient in sterilization. These results suggest that Chloride Dioxide (ClO2) and high concentration of CO2 gas injection treatments were an effective decontamination technique for improving the safety of strawberries.

Keywords: chloride dioxide, high concentration of CO2, modified atmosphere condition, oxygen transmission rate films

Procedia PDF Downloads 338
326 Effects of Plasma Technology in Biodegradable Films for Food Packaging

Authors: Viviane P. Romani, Bradley D. Olsen, Vilásia G. Martins

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Biodegradable films for food packaging have gained growing attention due to environmental pollution caused by synthetic films and the interest in the better use of resources from nature. Important research advances were made in the development of materials from proteins, polysaccharides, and lipids. However, the commercial use of these new generation of sustainable materials for food packaging is still limited due to their low mechanical and barrier properties that could compromise the food quality and safety. Thus, strategies to improve the performance of these materials have been tested, such as chemical modifications, incorporation of reinforcing structures and others. Cold plasma is a versatile, fast and environmentally friendly technology. It consists of a partially ionized gas containing free electrons, ions, and radicals and neutral particles able to react with polymers and start different reactions, leading to the polymer degradation, functionalization, etching and/or cross-linking. In the present study, biodegradable films from fish protein prepared through the casting technique were plasma treated using an AC glow discharge equipment. The reactor was preliminary evacuated to ~7 Pa and the films were exposed to air plasma for 2, 5 and 8 min. The films were evaluated by their mechanical and water vapor permeability (WVP) properties and changes in the protein structure were observed using Scanning Electron Microscopy (SEM) and X-ray diffraction (XRD). Potential cross-links and elimination of surface defects by etching might be the reason for the increase in tensile strength and decrease in the elongation at break observed. Among the times of plasma application tested, no differences were observed when higher times of exposure were used. The X-ray pattern showed a broad peak at 2θ = 19.51º that corresponds to the distance of 4.6Å by applying the Bragg’s law. This distance corresponds to the average backbone distance within the α-helix. Thus, the changes observed in the films might indicate that the helical configuration of fish protein was disturbed by plasma treatment. SEM images showed surface damage in the films with 5 and 8 min of plasma treatment, indicating that 2 min was the most adequate time of treatment. It was verified that plasma removes water from the films once weight loss of 4.45% was registered for films treated during 2 min. However, after 24 h in 50% of relative humidity, the water lost was recovered. WVP increased from 0.53 to 0.65 g.mm/h.m².kPa after plasma treatment during 2 min, that is desired for some foods applications which require water passage through the packaging. In general, the plasma technology affects the properties and structure of fish protein films. Since this technology changes the surface of polymers, these films might be used to develop multilayer materials, as well as to incorporate active substances in the surface to obtain active packaging.

Keywords: fish protein films, food packaging, improvement of properties, plasma treatment

Procedia PDF Downloads 161
325 Influence of Microparticles in the Contact Region of Quartz Sand Grains: A Micro-Mechanical Experimental Study

Authors: Sathwik Sarvadevabhatla Kasyap, Kostas Senetakis

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The mechanical behavior of geological materials is very complex, and this complexity is related to the discrete nature of soils and rocks. Characteristics of a material at the grain scale such as particle size and shape, surface roughness and morphology, and particle contact interface are critical to evaluate and better understand the behavior of discrete materials. This study investigates experimentally the micro-mechanical behavior of quartz sand grains with emphasis on the influence of the presence of microparticles in their contact region. The outputs of the study provide some fundamental insights on the contact mechanics behavior of artificially coated grains and can provide useful input parameters in the discrete element modeling (DEM) of soils. In nature, the contact interfaces between real soil grains are commonly observed with microparticles. This is usually the case of sand-silt and sand-clay mixtures, where the finer particles may create a coating on the surface of the coarser grains, altering in this way the micro-, and thus the macro-scale response of geological materials. In this study, the micro-mechanical behavior of Leighton Buzzard Sand (LBS) quartz grains, with interference of different microparticles at their contact interfaces is studied in the laboratory using an advanced custom-built inter-particle loading apparatus. Special techniques were adopted to develop the coating on the surfaces of the quartz sand grains so that to establish repeatability of the coating technique. The characterization of the microstructure of coated particles on their surfaces was based on element composition analyses, microscopic images, surface roughness measurements, and single particle crushing strength tests. The mechanical responses such as normal and tangential load – displacement behavior, tangential stiffness behavior, and normal contact behavior under cyclic loading were studied. The behavior of coated LBS particles is compared among different classes of them and with pure LBS (i.e. surface cleaned to remove any microparticles). The damage on the surface of the particles was analyzed using microscopic images. Extended displacements in both normal and tangential directions were observed for coated LBS particles due to the plastic nature of the coating material and this varied with the variation of the amount of coating. The tangential displacement required to reach steady state was delayed due to the presence of microparticles in the contact region of grains under shearing. Increased tangential loads and coefficient of friction were observed for the coated grains in comparison to the uncoated quartz grains.

Keywords: contact interface, microparticles, micro-mechanical behavior, quartz sand

Procedia PDF Downloads 186
324 COVID-19: Potential Effects of Nutritional Factors on Inflammation Relief

Authors: Maryam Nazari

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COVID-19 is a respiratory disease triggered by the novel coronavirus, SARS-CoV-2, that has reached pandemic status today. Acute inflammation and immune cells infiltration into lung injuries result in multi-organ failure. The presence of other non-communicable diseases (NCDs) with systemic inflammation derived from COVID-19 may exacerbate the patient's situation and increase the risk for adverse effects and mortality. This pandemic is a novel situation and the scientific community at this time is looking for vaccines or drugs to treat the pathology. One of the biggest challenges is focused on reducing inflammation without compromising the correct immune response of the patient. In this regard, addressing the nutritional factors should not be overlooked not only as a matter of avoiding the presence of NCDs with severe infections but also as an adjunctive way to modulate the inflammatory status of the patients. Despite the pivotal role of nutrition in modifying immune response, due to the novelty of the COVID-19 disease, information about the effects of specific dietary agents is limited in this area. From the macronutrients point of view, protein deficiency (quantity or quality) has negative effects on the number of functional immunoglobulins and gut-associated lymphoid tissue (GALT). High biological value proteins or some amino acids like arginine and glutamine are well known for their ability to augment the immune system. Among lipids, fish oil has the ability to inactivate enveloped viruses, suppress pro-inflammatory prostaglandin production and block platelet-activating factors and their receptors. In addition, protectin D1, which is an Omega-3 PUFAs derivation, is a novel antiviral drug. So it seems that these fatty acids can reduce the severity and/or improve recovery of patients with COVID-19. Carbohydrates with lower glycemic index and fibers are associated with lower levels of inflammatory cytokines (CRP, TNF-α, and IL-6). Short-Chain Fatty acids not only exert a direct anti-inflammatory effect but also provide appropriate gut microbial, which is important in gastrointestinal issues related to COVID-19. From the micronutrients point of view, Vitamins A, C, D, E, iron, magnesium, zinc, selenium and copper play a vital role in the maintenance of immune function. Inadequate status in these nutrients may result in decreased resistance against COVID-19 infection. There are specific bioactive compounds in the diet that interact with the ACE2 receptor, which is the gateway for SARS and SARS-CoV-2, and thus controls the viral infection. Regarding this, the potential benefits of probiotics, resveratrol (a polyphenol found in grape), oleoylethanolamide (derived from oleic acid), and natural peroxisome proliferator-activated receptor γ agonists in foodstuffs (like curcumin, pomegranate, hot pepper) are suggested. Yet, it should be pointed out that most of these results have been reported in animal models and further human studies are needed to be verified.

Keywords: Covid-19, inflammation, nutrition, dietary agents

Procedia PDF Downloads 173
323 Energy Metabolism and Mitochondrial Biogenesis in Muscles of Rats Subjected to Cold Water Immersion

Authors: Bosiacki Mateusz, Anna Lubkowska, Dariusz Chlubek, Irena Baranowska-Bosiacka

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Exposure to cold temperatures can be considered a stressor that can lead to adaptive responses. The present study hypothesized the possibility of a positive effect of cold water exercise on mitochondrial biogenesis and muscle energy metabolism in aging rats. The purpose of this study was to evaluate the effects of cold water exercise on energy status, purine compounds, and mitochondrial biogenesis in the muscles of aging rats as indicators of the effects of cold water exercise and their usefulness in monitoring adaptive changes. The study was conducted on 64 aging rats of both sexes, 15 months old at the time of the experiment. The rats (male and female separately) were randomly assigned to the following study groups: control, sedentary animals; 5°C groups animals - training swimming in cold water at 5°C; 36°C groups - animals training swimming in water at thermal comfort temperature. The study was conducted with the approval of the Local Ethical Committee for Animal Experiments. The animals in the experiment were subjected to swimming training for 9 weeks. During the first week of the study, the duration of the first swimming training was 2 minutes (on the first day), increasing daily by 0.5 minutes up to 4 minutes on the fifth day of the first week. From the second to the eighth week, the swimming training was 4 minutes per day, five days a week. At the end of the study, forty-eight hours after the last swim training, the animals were dissected. In the skeletal muscle tissue of the thighs of the rats, we determined the concentrations of ATP, ADP, AMP, Ado (HPLC), PGC-1a protein expression (Western blot), PGC1A, Mfn1, Mfn2, Opa1, and Drp1 gene expression (qRT PCR). The study showed that swimming in water at a thermally comfortable temperature improved the energy metabolism of the aging rat muscles by increasing the metabolic rate (increase in ATP, ADP, TAN, AEC) and enhancing mitochondrial fusion (increase in mRNA expression of regulatory proteins Mfn1 and Mfn2). Cold water swimming improved muscle energy metabolism in aging rats by increasing the rate of muscle energy metabolism (increase in ATP, ADP, TAN, AEC concentrations) and enhancing mitochondrial biogenesis and dynamics (increase in the mRNA expression of proteins of fusion-regulating factors – Mfn1, Mfn2, and Opa1, and the factor regulating mitochondrial fission – Drp1). The concentration of high-energy compounds and the expression of proteins regulating mitochondrial dynamics in the muscle may be a useful indicator in monitoring adaptive changes occurring in aging muscles under the influence of exercise in cold water. It represents a short-term adaptation to changing environmental conditions and has a beneficial effect on maintaining the bioenergetic capacity of muscles in the long term. Conclusion: exercise in cold water can exert positive effects on energy metabolism, biogenesis and dynamics of mitochondria in aging rat muscles. Enhancement of mitochondrial dynamics under cold water exercise conditions can improve mitochondrial function and optimize the bioenergetic capacity of mitochondria in aging rat muscles.

Keywords: cold water immersion, adaptive responses, muscle energy metabolism, aging

Procedia PDF Downloads 74
322 The Influences of Facies and Fine Kaolinite Formation Migration on Sandstone's Reservoir Quality, Sarir Formation, Sirt Basin Libya

Authors: Faraj M. Elkhatri

Abstract:

The spatial and temporal distribution of diagenetic alterations related impact on the reservoir quality of the Sarir Formation. ( present day burial depth of about 9000 feet) Depositional facies and diagenetic alterations are the main controls on reservoir quality of Sarir Formation Sirt Basin Libya; these based on lithology and grain size as well as authigenic clay mineral types and their distributions. However, petrology investigation obtained on study area with five sandstone wells concentrated on main rock components and the parameters that may have impacts on reservoirs. the main authigenic clay minerals are kaolinite and dickite, these investigations have confirmed by X.R.D analysis and clay fraction. mainly Kaolinite and Dickite were extensively presented on all of wells with high amounts. As well as trace of detrital smectite and less amounts of illitized mud-matrix are possibly find by SEM image. Thin layers of clay presented as clay-grain coatings in local depth interpreted as remains of dissolved clay matrix is partly transformed into kaolinite adjacent and towards pore throat. This also may have impacts on most of the pore throats of this sandstone which are open and relatively clean with some fine martial have been formed on occluded pores. This material is identified by EDS analysis to be collections of not only kaolinite booklets but also small disaggregated kaolinite platelets derived from the disaggregation of larger kaolinite booklets. These patches of kaolinite not only fill this pore but also coat some of the surrounding framework grains. Quartz grains often enlarged by authigenic quartz overgrowths partially occlude and reduce porosity. Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM) was conducted on the post-test samples to examine any mud filtrate particles that may be in the pore throats. Semi-qualitative elemental data on selected minerals observed during the SEM study were obtained through the use of an Energy Dispersive Spectroscopy (EDS) unit. The samples showed mostly clean open pore throats with limited occlusion by kaolinite. very fine-grained elemental combinations (Si/Al/Na/Cl, Si/Al Ca/Cl/Ti, and Qtz/Ti) have been identified and conformed by EDS analysis. However, the identification of the fine grained disaggregated material as mainly kaolinite though study area.

Keywords: pore throat, fine migration, formation damage, solids plugging, porosity loss

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321 Investigation on Correlation of Earthquake Intensity Parameters with Seismic Response of Reinforced Concrete Structures

Authors: Semra Sirin Kiris

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Nonlinear dynamic analysis is permitted to be used for structures without any restrictions. The important issue is the selection of the design earthquake to conduct the analyses since quite different response may be obtained using ground motion records at the same general area even resulting from the same earthquake. In seismic design codes, the method requires scaling earthquake records based on site response spectrum to a specified hazard level. Many researches have indicated that this limitation about selection can cause a large scatter in response and other charecteristics of ground motion obtained in different manner may demonstrate better correlation with peak seismic response. For this reason influence of eleven different ground motion parameters on the peak displacement of reinforced concrete systems is examined in this paper. From conducting 7020 nonlinear time history analyses for single degree of freedom systems, the most effective earthquake parameters are given for the range of the initial periods and strength ratios of the structures. In this study, a hysteresis model for reinforced concrete called Q-hyst is used not taken into account strength and stiffness degradation. The post-yielding to elastic stiffness ratio is considered as 0.15. The range of initial period, T is from 0.1s to 0.9s with 0.1s time interval and three different strength ratios for structures are used. The magnitude of 260 earthquake records selected is higher than earthquake magnitude, M=6. The earthquake parameters related to the energy content, duration or peak values of ground motion records are PGA(Peak Ground Acceleration), PGV (Peak Ground Velocity), PGD (Peak Ground Displacement), MIV (Maximum Increamental Velocity), EPA(Effective Peak Acceleration), EPV (Effective Peak Velocity), teff (Effective Duration), A95 (Arias Intensity-based Parameter), SPGA (Significant Peak Ground Acceleration), ID (Damage Factor) and Sa (Spectral Response Spectrum).Observing the correlation coefficients between the ground motion parameters and the peak displacement of structures, different earthquake parameters play role in peak displacement demand related to the ranges formed by the different periods and the strength ratio of a reinforced concrete systems. The influence of the Sa tends to decrease for the high values of strength ratio and T=0.3s-0.6s. The ID and PGD is not evaluated as a measure of earthquake effect since high correlation with displacement demand is not observed. The influence of the A95 is high for T=0.1 but low related to the higher values of T and strength ratio. The correlation of PGA, EPA and SPGA shows the highest correlation for T=0.1s but their effectiveness decreases with high T. Considering all range of structural parameters, the MIV is the most effective parameter.

Keywords: earthquake parameters, earthquake resistant design, nonlinear analysis, reinforced concrete

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320 An Analytical Metric and Process for Critical Infrastructure Architecture System Availability Determination in Distributed Computing Environments under Infrastructure Attack

Authors: Vincent Andrew Cappellano

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In the early phases of critical infrastructure system design, translating distributed computing requirements to an architecture has risk given the multitude of approaches (e.g., cloud, edge, fog). In many systems, a single requirement for system uptime / availability is used to encompass the system’s intended operations. However, when architected systems may perform to those availability requirements only during normal operations and not during component failure, or during outages caused by adversary attacks on critical infrastructure (e.g., physical, cyber). System designers lack a structured method to evaluate availability requirements against candidate system architectures through deep degradation scenarios (i.e., normal ops all the way down to significant damage of communications or physical nodes). This increases risk of poor selection of a candidate architecture due to the absence of insight into true performance for systems that must operate as a piece of critical infrastructure. This research effort proposes a process to analyze critical infrastructure system availability requirements and a candidate set of systems architectures, producing a metric assessing these architectures over a spectrum of degradations to aid in selecting appropriate resilient architectures. To accomplish this effort, a set of simulation and evaluation efforts are undertaken that will process, in an automated way, a set of sample requirements into a set of potential architectures where system functions and capabilities are distributed across nodes. Nodes and links will have specific characteristics and based on sampled requirements, contribute to the overall system functionality, such that as they are impacted/degraded, the impacted functional availability of a system can be determined. A machine learning reinforcement-based agent will structurally impact the nodes, links, and characteristics (e.g., bandwidth, latency) of a given architecture to provide an assessment of system functional uptime/availability under these scenarios. By varying the intensity of the attack and related aspects, we can create a structured method of evaluating the performance of candidate architectures against each other to create a metric rating its resilience to these attack types/strategies. Through multiple simulation iterations, sufficient data will exist to compare this availability metric, and an architectural recommendation against the baseline requirements, in comparison to existing multi-factor computing architectural selection processes. It is intended that this additional data will create an improvement in the matching of resilient critical infrastructure system requirements to the correct architectures and implementations that will support improved operation during times of system degradation due to failures and infrastructure attacks.

Keywords: architecture, resiliency, availability, cyber-attack

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319 Effect of Endurance Training on Serum Chemerin Levels and Lipid Profile of Plasma in Obese Women

Authors: A. Moghadasein, M. Ghasemi, S. Fazelifar

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Aim: Chemerin is a novel adipokine that play an important role in regulating lipid metabolism and abiogenesis. Chemerin is dependent on autocrine and paracrine signals for the differentiation and maturation of fat cells; it also regulates glucose uptake in fat cells and stimulates lipolysis. It has been reported that in adipocytes, chemerin enhances the insulin-stimulated glucose and causes the phosphorylation of tyrosine in Insulin receptor substrate. According to the studies, Chemerin may increase insulin sensitivity in adipose tissue and is largely associated with Body mass index, triglycerides, and blood pressure in those with normal glucose tolerance. There is limited information available regarding the effect of exercise training on serum chemerin concentrations. The purpose of this study was to investigate the effect of endurance training on serum chemerin levels and lipids of plasma in overweight women. Methodology: This study was a quasi-experimental research with a pre-post test design. After required examination and verification of high pressure by the physician, 22 obese subjects (age: 35.64±5.55 yr, weight: 75.62±9.30 kg, body mass index: 32.4±1.6 kg/m2) were randomly assigned to aerobic training (n= 12) and control (n= 12) groups. Participants completed a questionnaire indicating the lack of sports history during the past six months, the lack of anti-hypertension drugs use, hormone therapy, cardiovascular problems, and complete stoppage of menstrual cycle. Aerobic training was performed 3 times weekly for 8 weeks. Resting levels of chemerin plasma, metabolic parameters were measured prior to and after the intervention. The control group did not participate in any training program. In this study, ethical considerations included the complete description of the objectives to the study participants, ensuring the confidentiality of their information. Kolmogorov-Smirnov and Levin test were used for determining the normal distribution of data and homogeneity of variances, respectively. Analyze of variance with repeated measure were used to investigate the changes in the intra-group and the differences in inter-group of variables. Statistical operations were performed using SPSS 16 and the significance level of the tests was considered at P < 0.05. Results: After an 8 week aerobic training, levels of chemerin plasma were significantly decreased in aerobic trained group when compared with their control groups (p < 0.05).Concurrently, levels of HDL-c were significantly decreased (p < 0.05) whereas, levels of cholesterol, TG and LDL-c, showed no significant changes (p > 0.05). No significant correlations between chemerin levels and weight loss were observed in subjects with overweight women. Conclusion: The present study demonstrated, 8 weeks aerobic training, reduced serum chemerin concentrations in overweight women. Whereas, aerobic training exercise programmers affected the lipid profile response of obese subjects differently. However further research is warranted in order to unravel the molecular mechanism for the range of responses and the role of serum chemerin.

Keywords: chemerin, aerobic training, lipid profile, obese women

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318 The Potential Fresh Water Resources of Georgia and Sustainable Water Management

Authors: Nana Bolashvili, Vakhtang Geladze, Tamazi Karalashvili, Nino Machavariani, George Geladze, Davit Kartvelishvili, Ana Karalashvili

Abstract:

Fresh water is the major natural resource of Georgia. The average perennial sum of the rivers' runoff in Georgia is 52,77 km³, out of which 9,30 km³ inflows from abroad. The major volume of transit river runoff is ascribed to the Chorokhi river. Average perennial runoff in Western Georgia is 41,52 km³, in Eastern Georgia 11,25 km³. The indices of Eastern and Western Georgia were calculated with 50% and 90% river runoff respectively, while the same index calculation for other countries is based on a 50% river runoff. Out of total volume of resources, 133,2 m³/sec (4,21 km³) has been geologically prospected by the State Commission on Reserves and Acknowledged as reserves available for exploitation, 48% (2,02 km³) of which is in Western Georgia and 2,19 km³ in Eastern Georgia. Considering acknowledged water reserves of all categories per capita water resources accounts to 2,2 m³/day, whereas high industrial category -0. 88 m³ /day fresh drinking water. According to accepted norms, the possibility of using underground water reserves is 2,5 times higher than the long-term requirements of the country. The volume of abundant fresh-water reserves in Georgia is about 150 m³/sec (4,74 km³). Water in Georgia is consumed mostly in agriculture for irrigation purposes. It makes 66,4% around Georgia, in Eastern Georgia 72,4% and 38% in Western Georgia. According to the long-term forecast provision of population and the territory with water resources in Eastern Georgia will be quite normal. A bit different is the situation in the lower reaches of the Khrami and Iori rivers which could be easily overcome by corresponding financing. The present day irrigation system in Georgia does not meet the modern technical requirements. The overall efficiency of their majority varies between 0,4-0,6. Similar is the situation in the fresh water and public service water consumption. Organization of the mentioned systems, installation of water meters, introduction of new methods of irrigation without water loss will substantially increase efficiency of water use. Besides new irrigation norms developed from agro-climatic, geographical and hydrological angle will significantly reduce water waste. Taking all this into account we assume that for irrigation agricultural lands in Georgia is necessary 6,0 km³ water, 5,5 km³ of which goes to Eastern Georgia on irrigation arable areas. To increase water supply in Eastern Georgian territory and its population is possible by means of new water reservoirs as the runoff of every river considerably exceeds the consumption volume. In conclusion, we should say that fresh water resources by which Georgia is that rich could be significant source for barter exchange and investment attraction. Certain volume of fresh water can be exported from Western Georgia quite trouble free, without bringing any damage to population and hydroecosystems. The precise volume of exported water per region/time and method/place of water consumption should be defined after the estimation of different hydroecosystems and detailed analyses of water balance of the corresponding territories.

Keywords: GIS, management, rivers, water resources

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317 Enzymatic Determination of Limonene in Red Clover Genotypes

Authors: Andrés Quiroz, Emilio Hormazabal, Ana Mutis, Fernando Ortega, Manuel Chacón-Fuentes, Leonardo Parra

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Red clover (Trifolium pratense L.) is an important forage species in temperate regions of the world. The main limitation of this species worldwide is a lack of persistence related to the high mortality of plants due to a complex of biotic and abiotic factors, determining a life span of two or three seasons. Because of the importance of red clover in Chile, a red clover breeding program was started at INIA Carillanca Research Center in 1989, with the main objective of improving the survival of plants, forage yield, and persistence. The main selection criteria for selecting new varieties have been based on agronomical parameters and biotic factors. The main biotic factor associated with red clover mortality in Chile is Hylastinus obscurus (Coleoptera: Curculionidae). Both larval and adults feed on the roots, causing weakening and subsequent death of clover plants. Pesticides have not been successful for controlling infestations of this root borer. Therefore, alternative strategies for controlling this pest are a high priority for red clover producers. Currently, the role of semiochemical in the interaction between H. obscurus and red clover plants has been widely studied for our group. Specifically, from the red clover foliage has been identified limonene is eliciting repellency from the root borer. Limonene is generated in the plant from two independent biosynthetic pathways, the mevalonic acid, and deoxyxylulose pathway. Mevalonate pathway enzymes are localized in the cytosol, whereas the deoxyxylulose phosphate pathway enzymes are found in plastids. In summary, limonene can be determinated by enzymatic bioassay using GPP as substrate and by limonene synthase expression. Therefore, the main objective of this work was to study genetic variation of limonene in material provided by INIA´s Red Clover breeding program. Protein extraction was carried out homogenizing 250 mg of leave tissue and suspended in 6 mL of extraction buffer (PEG 1500, PVP-30, 20 mM MgCl2 and antioxidants) and stirred on ice for 20 min. After centrifugation, aliquots of 2.5 mL were desalted on PD-10 columns, resulting in a final volume of 3.5 mL. Protein determination was performed according to Bradford with BSA as a standard. Monoterpene synthase assays were performed with 50 µL of protein extracts transferred into gas-tight 2 mL crimp seal vials after addition of 4 µL MgCl₂ and 41 µL assay buffer. The assay was started by adding 5 µL of a GPP solution. The mixture was incubated for 30 min at 40 °C. Biosynthesized limonene was quantified in a GC equipped with a chiral column and using synthetic R and S-limonene standards. The enzymatic the production of R and S-limonene from different Superqueli-Carillanca genotypes is shown in this work. Preliminary results showed significant differences in limonene content among the genotypes analyzed. These results constitute an important base for selecting genotypes with a high content of this repellent monoterpene towards H. obscurus.

Keywords: head space, limonene enzymatic determination, red clover, Hylastinus obscurus

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