Search results for: extracellular matrix mineralization

Authors: D. Adel, F. Giacomini, R. Gildenhaar, G. Berger, C. Gomes, U. Linow, M. Hardt, B. Peleskae, J. Günster, A. Houshmand, M. Stiller, A. Rack, K. Ghaffar, A. Gamal, M. El Mofty, C. Knabe

Abstract:

The goal of this study was to develop 3D scaffolds from a silica containing calcium alkali orthophosphate utilizing two different fabrication processes, first a replica technique namely the Schwartzwalder Somers method (SSM), and second 3D printing, i.e. Rapid prototyping (RP). First, the mechanical and physical properties of the scaffolds (porosity, compressive strength, and solubility) was assessed and second their potential to facilitate homogenous colonization with osteogenic cells and extracellular bone matrix formation throughout the porous scaffold architecture. To this end murine and rat calavarie osteoblastic cells were dynamically seeded on both scaffold types under perfusion with concentrations of 3 million cells. The amount of cells and extracellular matrix as well as osteogenic marker expression was evaluated using hard tissue histology, immunohistochemistry, and histomorphometric analysis. Total porosities of both scaffolds were 86.9 % and 50% for SSM and RP respectively, Compressive strength values were 0.46 ± 0.2 MPa for SSM and 6.6± 0.8 MPa for RP. Regarding the cellular behavior, RP scaffolds displayed a higher cell and matrix percentage of 24.45%. Immunoscoring yielded strong osteocalcin expression of cells and matrix in RP scaffolds and a moderate expression in SSM scaffolds. 3D printed RP scaffolds displayed superior mechanical and biological properties compared to SSM. 3D printed scaffolds represent excellent candidates for bone tissue engineering.

Keywords: calcium alkali orthophosphate, extracellular matrix mineralization, osteoblast differentiation, rapid prototyping, scaffold

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Authors: Hawon Lee, Young-Pil Kim

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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.

Keywords: collagen, ECM, FRET, MMP

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Authors: Yun-An Chen, Hung-Jun Lin, Tai-Horng Young, Der-Zen Liu

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Decellularized brain extracellular matrix had been shown that it has the ability to influence on cell proliferation, differentiation and associated cell phenotype. However, this scaffold is thought to have poor mechanical properties and rapid degradation, it is hard for cell recellularization. In this study, we used decellularized brain extracellular matrix combined with chitosan, which is naturally occurring polysaccharide and non-cytotoxic polymer, forming a 3-D scaffold for neural stem/precursor cells (NSPCs) regeneration. HE staining and DAPI fluorescence staining confirmed decellularized process could effectively vanish the cellular components from the brain. GAGs and collagen I, collagen IV were be showed a great preservation by Alcain staining and immunofluorescence staining respectively. Decellularized brain extracellular matrix was well mixed in chitosan to form a 3-D scaffold (DB-C scaffold). The pore size was approximately 50±10 μm examined by SEM images. Alamar blue results demonstrated NSPCs had great proliferation ability in DB-C scaffold. NSPCs that were cultured in this complex scaffold differentiated into neurons and astrocytes, as reveled by NSPCs expression of microtubule-associated protein 2 (MAP2) and glial fibrillary acidic protein (GFAP). In conclusion, DB-C scaffold may provide bioinformatics cues for NSPCs generation and aid for CNS injury functional recovery applications.

Keywords: brain, decellularization, chitosan, scaffold, neural stem/precursor cells

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Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

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The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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Authors: Valeria Panzetta, Ida Musella, Sabato Fusco, Paolo Antonio Netti

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The study of the biophysics of living cells drew attention to the pivotal role of the cytoskeleton in many cell functions, such as mechanics, adhesion, proliferation, migration, differentiation and neoplastic transformation. In particular, during the complex process of malignant transformation and invasion cell cytoskeleton devolves from a rigid and organized structure to a more compliant state, which confers to the cancer cells a great ability to migrate and adapt to the extracellular environment. In order to better understand the malignant transformation process from a mechanical point of view, it is necessary to evaluate the direct crosstalk between the cells and their surrounding extracellular matrix (ECM) in a context which is close to in vivo conditions. In this study, human biopsy tissues of lung adenocarcinoma were analyzed in order to define their mechanical phenotype at cell and ECM level, by using particle tracking microrheology (PTM) technique. Polystyrene beads (500 nm) were introduced into the sample slice. The motion of beads was obtained by tracking their displacements across cell cytoskeleton and ECM structures and mean squared displacements (MSDs) were calculated from bead trajectories. It has been already demonstrated that the amplitude of MSD is inversely related to the mechanical properties of intracellular and extracellular microenvironment. For this reason, MSDs of particles introduced in cytoplasm and ECM of healthy and tumor tissues were compared. PTM analyses showed that cancerous transformation compromises mechanical integrity of cells and extracellular matrix. In particular, the MSD amplitudes in cells of adenocarcinoma were greater as compared to cells of normal tissues. The increased motion is probably associated to a less structured cytoskeleton and consequently to an increase of deformability of cells. Further, cancer transformation is also accompanied by extracellular matrix stiffening, as confirmed by the decrease of MSDs of matrix in tumor tissue, a process that promotes tumor proliferation and invasiveness, by activating typical oncogenic signaling pathways. In addition, a clear correlation between MSDs of cells and tumor grade was found. MSDs increase when tumor grade passes from 2 to 3, indicating that cells undergo to a trans-differentiation process during tumor progression. ECM stiffening is not dependent on tumor grade, but the tumor stage resulted to be strictly correlated with both cells and ECM mechanical properties. In fact, a greater stage is assigned to tumor spread to regional lymph nodes and characterized by an up-regulation of different ECM proteins, such as collagen I fibers. These results indicate that PTM can be used to get nanomechanical characterization at different scale levels in an interpretative and diagnostic context.

Keywords: cytoskeleton, extracellular matrix, mechanical properties, particle tracking microrheology, tumor

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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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Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: extracellular laccase, production, yeast, natural inducer

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Authors: Sandip Sharma, Jayesh Ruparelia

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The present work focuses on the comparative study of ozone based advanced oxidation processes (AOPs): O3, O3/UV and O3/UV/Persulfate for mineralization of synthetic wastewater containing Reactive Black5 (RB5) dye. The effect of various parameters: pH, ozone flow rate, initial concentration of dye and intensity of UV light was analyzed to access performance efficiency of AOPs. The performance of all the three AOPs was evaluated on the basis of decolorization, % TOC removal and ozone consumption. The highest mineralization rate of 86.83% was achieved for O3/UV/Persulfate followed by 71.53% and 66.82 % for O3/UV and O3 respectively. This is attributed to the fact that Persulfate ions (S2O82-) upon activation produce sulfate radical (SO4-●) which is very strong oxidant capable of degrading a wide variety of recalcitrant organic compounds, moreover to enhance the performance of Persulfate it is activated using UV irradiation. On increasing the intensity of UV irradiation from 11W to 66W, TOC removal efficiency is increased by 59.04%. Ozone based AOPs gives better mineralization on basic conditions, at pH 12 it gives 68.81%, 60.01% and 40.32% TOC removal for O3/UV/Persulfate, O3/UV and O3 process respectively. The result also reveals that decolorization of 98.95%, 95.17% and 94.71% was achieved by O3/UV/Persulfate, O3/UV and O3 process respectively. In addition to above, ozone consumption was also considerably decreased by 17% in case of O3/UV/Persulfate, as efficiency of process is enhanced by means of activation of persulfate through UV irradiation. Thus study reveals that mineralization follows: O3/UV/Persulfate> O3/UV> O3.

Keywords: AOP, mineralization, TOC, recalcitrant organic compounds

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Authors: Seok Woo Chang

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Aim: Capseal I and Capseal II are calcium silicate and calcium phosphate based experimental root canal sealer. The aim of this study was to evaluate the biocompatibility and mineralization potential of Capseal I and Capseal II. Materials and Methods: The biocompatibility and mineralization-related gene expression (alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN)) of Capseal I and Capseal II were compared using methylthiazol tetrazolium assay and reverse transcription-polymerization chain reaction analysis, respectively. The results were analyzed by Kruskal-Wallis test. P-value of < 0.05 was considered significant. Result: Both Capseal I and Capseal II were favorable in biocompatibility and influenced the messenger RNA expression of ALP and BSP. Conclusion: Within the limitation of this study, Capseal is biocompatible and have mineralization promoting potential, and thus could be a promising root canal sealer.

Keywords: biocompatibility, mineralization-related gene expression, Capseal I, Capseal II

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Authors: Hossein Bazmara, Kaamran Raahemifar, Mostafa Sefidgar, Madjid Soltani

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Angiogenesis is formation of new blood vessels from existing vessels. Due to flow of blood in vessels, during angiogenesis, blood flow plays an important role in regulating the angiogenesis process. Multiple mathematical models of angiogenesis have been proposed to simulate the formation of the complicated network of capillaries around a tumor. In this work, a multi-scale model of angiogenesis is developed to show the effect of blood flow on capillaries and network formation. This model spans multiple temporal and spatial scales, i.e. intracellular (molecular), cellular, and extracellular (tissue) scales. In intracellular or molecular scale, the signaling cascade of endothelial cells is obtained. Two main stages in development of a vessel are considered. In the first stage, single sprouts are extended toward the tumor. In this stage, the main regulator of endothelial cells behavior is the signals from extracellular matrix. After anastomosis and formation of closed loops, blood flow starts in the capillaries. In this stage, blood flow induced signals regulate endothelial cells behaviors. In cellular scale, growth and migration of endothelial cells is modeled with a discrete lattice Monte Carlo method called cellular Pott's model (CPM). In extracellular (tissue) scale, diffusion of tumor angiogenic factors in the extracellular matrix, formation of closed loops (anastomosis), and shear stress induced by blood flow is considered. The model is able to simulate the formation of a closed loop and its extension. The results are validated against experimental data. The results show that, without blood flow, the capillaries are not able to maintain their integrity.

Keywords: angiogenesis, endothelial cells, multi-scale model, cellular Pott's model, signaling cascade

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Authors: Mohammad Salem Abdullah Al-Hwaiti

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The stratiform copper mineralization of the Burj-Dolomite shale (BDS) formations of deposits shows that the copper mineralization within the BDS occurs as hydrated copper chlorides and carbonates (mainly paratacamite and malachite, respectively), while copper silicates (mainly chrysocolla and planchette) are the major ore minerals in the BDS. Thus, on the basis of the petrographic and field occurrence, three main stages operated during the development of the copper ore in the sandy and shaly lithofacies. During the first stage, amorphous chrysocolla replaced clays, feldspars, and quartz. This stage was followed by the transition from an amorphous phase to a better-crystallized phase, i.e., the formation of planchette and veins from chrysocolla. The third stage was the formation of chrysocolla along fracture planes. Other secondary minerals are pseudomalachite, dioptase, neoticite together with authigenic fluorapatite. Paratacamite and malachite, which are common in the dolomitic lithofacies, are relatively rare in the sandy and silty lithofacies. The Rare Earth Elements (REEs) patterns for the BDS showed three stages in the evolution of the Precambrian–Cambrian copper mineralization system, involving the following: (A) Epigenetic mobilization of Cu-bearing solution with formation Cu-carbonate in dolomite and limestone mineralization and Cu-silicate mineralization in sandstone; (B) Transgression of Cambrian Sea and SSC deposition of Cu-sulphides during dolomite diagenesis in the BDS Formation; continued diagenesis and oxidation leads to the formation of Cu(II) minerals; (C) Erosion and supergene enrichment of Cu in basement rocks. Detrital copper-bearing sediments accumulate in the lower Cambrian clastic sequence.

Keywords: dolomite shale, copper mineralization, REE, Jordan

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Authors: B. Merdas

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The Guelma region constitutes a vast geothermal field whose local geothermal gradient is very high. Indeed, various thermal and thermo sources emerging in the region, including some at relatively high temperatures. In the mio Pliocene Hammam N'bails, basin emerges a hot spring that leaves develop a thick series of thermal travertine linked to it. Near the thermal emergences has settled a very special mineralization antimony and zinc and lead. The results of analyses of the thermal waters of the source of Hammam N'bails and the associated travertine, show abnormal values in Pb, Sb, Zn, As, and other metals, demonstrating the genetic link between those waters and mineralization. Hammam N'bails mineralizations by their mineral assembling represented and their association with the hot springs, are very similar to epithermal deposits with precious metals (gold and silver) like Senator mine in Turkey or ‘Carlin-type’ in Nevada (USA).

Keywords: hot springs, mineralization; basement faults, Guelma, NE Algeria

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Authors: M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, A. K. Abdul Jalil, M. R. Salina, M. R. Maryam, M. Rosfarizan

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The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, streptomycin sulfate, extracellular proteins, antibiotics

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Authors: Alfi Y. Zakiyyah, Hanni Garminia, M. Salman, A. N. Irawati

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In the terminology of the hypergraph, there is a relation with the terminology graph. In the theory of graph, the edges connected two vertices. In otherwise, in hypergraph, the edges can connect more than two vertices. There is representation matrix of a graph such as adjacency matrix, Laplacian matrix, and incidence matrix. The adjacency matrix is symmetry matrix so that all eigenvalues is real. This matrix is a nonnegative matrix. The all diagonal entry from adjacency matrix is zero so that the trace is zero. Another representation matrix of the graph is the Laplacian matrix. Laplacian matrix is symmetry matrix and semidefinite positive so that all eigenvalues are real and non-negative. According to the spectral study in the graph, some that result is generalized to hypergraph. A hypergraph can be represented by a matrix such as adjacency, incidence, and Laplacian matrix. Throughout for this term, we use Laplacian matrix to represent a complete tripartite hypergraph. The aim from this research is to determine second smallest eigenvalues from this matrix and find a relation this eigenvalue with the connectivity of that hypergraph.

Keywords: connectivity, graph, hypergraph, Laplacian matrix

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Authors: Fakher Eddine Messaoudi

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The M'Taguinarou North iron occurrence contains Iron and polymetallic mineralization (Fe-Zn-Cu), hosted in Turonian limestone. It manifests in metric clusters of goethite and hematite and in centimetre veins of smithsonite, malachite, and azurite. The genesis of this mineralization is clearly polyphased and results from the supergene processes superposed on hydrothermal phases where the Triassic diapirs probably generated the circulation of hydrothermal fluids through the sedimentary series, and the alteration of the Turonian limestone gave the formation of the hydrothermal primary ore composed of iron carbonates (siderite). Several uplift episodes affected the mineralization and the host rocks, generating the genesis of a polymetallic supergene assembly (goethite, malachite, azurite, quartz, and calcite). In M’taguinarou North, iron oxy-hydroxides are mainly observed in the form of fibrous stalactites, stalagmites, and Botroydale structures, where hematite precipitated first, followed immediately by goethite, limonite, and smithsonite. Siderite is completely absent. Subsequently, malachite, azurite, and calcite formed in the form of small veins intersecting the surrounding limestone.

Keywords: mineralization, genetic model, hydrothermal iron, supergene, Tebessa, Algeria

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Authors: Prativa Das, Lay Poh Tan

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Three-dimensional microenvironment is a need to study the event cascades of liver fibrosis in vitro. Electrospun nanofibers modified with essential extracellular matrix proteins can closely mimic the random fibrous structure of native liver extracellular matrix (ECM). In this study, we fabricate a series of 3D electrospun scaffolds by wet electrospinning process modified with different ratios of collagen-I to fibronectin to achieve optimized distribution of these two ECM proteins on the fiber surface. A ratio of 3:1 of collagen-I to fibronectin was found to be optimum for surface modification of electrospun poly(lactic-co-glycolic acid) (PLGA) fibers by chemisorption process. In 3:1 collagen-I to fibronectin modified scaffolds the total protein content increased by ~2 fold compared to collagen-I modified and ~1.5 fold compared to 1:1/9:1 collagen-I to fibronectin modified scaffolds. We have cultured LX-2 cells on this scaffold over 14 days and found that LX-2 cells acquired more quiescent phenotype throughout the culture period and shown significantly lower expression of alpha smooth muscle actin and collagen-I. Thus, this system can be used as a model to study liver fibrosis by using different fibrogenic mediators in vitro.

Keywords: electrospinning, collagen-I and fibronectin, surface modification of fiber, LX-2 cells, liver fibrosis

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Authors: Ric Joseph R. Murillo, Edna N. Gueco, Dennis I. Merino

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Let F be C or R, where C and R are the set of complex numbers and real numbers, respectively, and n be a natural number. An n-by-n matrix A over the field F is called an α-involutory matrix or an α-involution if there exists an α in the field such that the square of the matrix is equal to αI, where I is the n-by-n identity matrix. If α is a complex number or a nonnegative real number, then an n-by-n matrix A over the field F can be written as a sum of n-by-n α-involutory matrices over the field F if and only if the trace of that matrix is an integral multiple of the square root of α. Meanwhile, if α is a negative real number, then a 2n-by-2n matrix A over R can be written as a sum of 2n-by-2n α-involutory matrices over R if and only the trace of the matrix is zero. Some other properties of α-involutory matrices are also determined

Keywords: α-involutory Matrices, sum of α-involutory Matrices, Trace, Matrix Theory

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Authors: Anna Cameron, Chunxia Zhao, Haofei Wang, Yun Liu, Guang Ze Yang

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Since the first publication in 1775, cancer research has built a comprehensive understanding of how cellular components of the tumor niche promote disease development. However, only within the last decade has research begun to establish the impact of non-cellular components of the niche, particularly the extracellular matrix (ECM). The ECM, a three-dimensional scaffold that sustains the tumor microenvironment, plays a crucial role in disease progression. Cancer cells actively deregulate and remodel the ECM to establish a tumor-promoting environment. Recent work has highlighted the need to further our understanding of the complexity of this cancer-ECM relationship. In vitro models use hydrogels to mimic the ECM, as hydrogel matrices offer biological compatibility and stability needed for long term cell culture. However, natural hydrogels are being used in these models verbatim, without tuning their biophysical characteristics to achieve pathophysiological relevance, thus limiting their broad use within cancer research. The biophysical attributes of these gels dictate cancer cell proliferation, invasion, metastasis, and therapeutic response. Evaluating the three most widely used natural hydrogels, Matrigel, collagen, and agarose gel, the permeability, stiffness, and pore-size of each gel were measured and compared to the in vivo environment. The pore size of all three gels fell between 0.5-6 µm, which coincides with the 0.1-5 µm in vivo pore size found in the literature. However, the stiffness for hydrogels able to support cell culture ranged between 0.05 and 0.3 kPa, which falls outside the range of 0.3-20,000 kPa reported in the literature for an in vivo ECM. Permeability was ~100x greater than in vivo measurements, due in large part to the lack of cellular components which impede permeation. Though, these measurements prove important when assessing therapeutic particle delivery, as the ECM permeability decreased with increasing particle size, with 100 nm particles exhibiting a fifth of the permeability of 10 nm particles. This work explores ways of adjusting the biophysical characteristics of hydrogels by changing protein concentration and the trade-off, which occurs due to the interdependence of these factors. The global aim of this work is to produce a more pathophysiologically relevant model for each tumor type.

Keywords: cancer, extracellular matrix, hydrogel, microfluidic

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Authors: Sourita Ghosh, Falguni Pati, Suhanya Duraiswamy

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Spheroid, a simple three-dimensional cellular aggregate, allows us to simulate the in-vivo complexity of cellular signaling and interactions in greater detail than traditional 2D cell culture. It can be used as an in-vitro model for drug toxicity testing, tumor modeling and many other such applications specifically for cancer. Our work is focused on the development of an affordable, user-friendly, robust, reproducible, high throughput microfluidic device for water in oil droplet production, which can, in turn, be used for spheroids manufacturing. Here, we have investigated the droplet breakup between two non-Newtonian fluids, viz. silicone oil and decellularized liver matrix, which acts as our extra cellular matrix (ECM) for spheroids formation. We performed some biochemical assays to characterize the liver ECM, as well as rheological studies on our two fluids and observed a critical dependence of capillary number (Ca) on droplet breakup and homogeneous drop formation

Keywords: chip less, droplets, extracellular matrix, liver spheroid

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Authors: Yu Wen Wang, Shyh Ming Kuo, Hsia Ying Cheng, Yu Chiuan Wu

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Liver fibrosis is caused by the activation of hepatic stellate cells in the liver to secrete excessive and deposition of extracellular matrix. In recent years, many treatment strategies have been developed to reduce the activation of hepatic stellate cells and therefore to increase the decomposition of extracellular matrix. Bacopa monnieri, an herbaceous plant of the scrophulariaceae, containing saponins and glycosides, which with antioxidant, anti-inflammation, pain relief and free radical scavenging characteristics. This study was to evaluate the inhibition of hepatic stellate cell activity by Bacopa monnieri extract and its therapeutic potential in treating thioacetamide-induced liver fibrosis in rats. The results showed that the IC50 of Bacopa monnieri extract was 0.39 mg/mL. Bacopa monnieri extract could effectively reduce H2O2-induced hepatic stellate cells inflammation. In the TAA-induced liver fibrosis animal studies, albumin secretion recovered to normal level after treated with Bacopa monnieri extract for 2-w, and fibrosis related proteins, α-SMA and TGF-1levels decreased indicating the extract exerted therapeutic effect on the liver fibrosis. However, inflammatory factors TNF- obviously decreased after 4-w treatment. In summary, we could successfully extract the main component-Bacopaside I from the plant and acquired a potential therapy using this component in treating TAA-induced liver fibrosis in rat.

Keywords: anti-inflammatory, Bacopa monnieri, fibrosis, hepatic stellate cells, water extract

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Authors: Ali Al-Bakri, Mohammed Sazid

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The investigated study area locates about 72 km from Jeddah city, Makkah district, Kingdom of Saudi Arabia. The study mainly aimed to define only in detail the most significant zones of possible mineralization and outline their subsurface parameters (location and strike) in the southeast part of Jabal Al-Qashah. Several geophysical methods have been conducted to carry out the goal. Among these methods are the ground magnetic method, self-potential (SP) method, and induced polarization (IP) method. Integrating these methods aims to help in delineating the possible mineralization in the study area. The magnetic survey was conducted along 17 profiles where these profiles were chosen to be perpendicular to the strike of the quartz shear zone. Self-potential was applied along with five profiles covering the study area. At the same time, induced polarization was used along with one profile located at the western side of the study area corresponding to some magnetic and SP profiles. The most interesting zones of mineralization were successfully determined by comparing the results of residual magnetic profile (3), SP profile (1), and IP profile, where geological structures control some mineralization.

Keywords: geophysical methods, magnetic method, self-potential, induced polarization, Jabal Al-Qashah

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Authors: Samir Verdiyev, Fuad Huseynov, Islam Guliyev, Coşqun İsmayıl

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The Filizchay polymetallic deposit is located on the southern slope of the Greater Caucasus Mountain Range, northwest of Azerbaijan in the Balaken district. Filizchay is the largest polymetallic deposit in the region and the second-largest polymetallic deposit in Europe. The mineral deposits in the region are associated with two different geodynamic evolutions that began with the Mesozoic collision along the Eurasian continent and the formation of a magmatic arc after the collision and continued with subduction in the Cenozoic. The bedrocks associated with Filizchay mineralization are Early Jurassic aged. The stratigraphic sequence of the deposit is consisting of black metamorphic clay shales, sandstones, and ore layers. Shales, sandstones, and siltstones are encountered in the upper and middle sections of the ore body, while only shales are observed at the lowest ranges. The ore body is mainly layered by the geometric structure of the bedrock; folding can be observed in the ore layers along with the bedrock foliation, and just in few points indirect laying due to the metamorphism. This suggests that the Filizchay ore mineralization is syngenetic, which is proved by the mineralization by the bedrock. To determine the ore petrography properties of the Filizchay deposit, samples were collected from the region where the ore is concentrated, and a polished section was prepared. These collected samples were examined under the mineralogical microscope to reveal the paragenesis of the mineralization and to explain the relation of ore minerals to each other. In this study, macroscopically observed minerals and textures of these minerals were used in the cores revealed during drilling exploration made by AzerGold CJS company. As a result of all these studies, it has been determined that there are three main mineralization types in the Filizchay deposit: banded, massive, and veinlet ores. The mineralization is in the massive pyrite; furthermore, the basis of the ore-mass contains pyrite, chalcopyrite, sphalerite, and galena. The pyrite in some parts of the ore body transformed to pyrrhotite as a result of metamorphism. Pyrite-chalcopyrite, pyrite-sphalerite-galena, pyrite-pyrrhotite mineral assemblages were determined during microscopic studies of mineralization. The replacement texture is more developed in Filizchay ores. The banded polymetallic type mineralization and near bedrocks are cut by quartz-carbonate veins. The geotectonic position and lithological conditions of the Filizchay deposit, the texture, and interrelationship of the sulfide mineralization indicate that it is a sedimentary-exhalative type of Au-Cu-Ag-Zn-Pb polymetallic deposit that is genetically related to the massive sulfide deposits.

Keywords: Balaken, Filizchay, metamorphism, polymetallic mineralization

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Authors: M. Hadji, N. Chiker, Y. Hadji, A. Haddad

Abstract:

In this paper, we report for the first time on the synthesis and characterization of novel MAX phases (Ti3SiC2, Ti2AlC) reinforced Ni-matrix and Ti2AlC reinforced Al-matrix. The stability of MAX phases in Al-matrix and Ni-matrix at a temperature of 985°C has been investigated. All the composites were cold pressed and sintered at a temperature of 985°C for 20min in H2 environment, except (Ni/Ti3SiC2) who was sintered at 1100°C for 1h.Microstructure analysis by scanning electron microscopy and phase analysis by X-Ray diffraction confirmed that there was minimal interfacial reaction between MAX particles and Ni, thus Al/MAX samples shown that MAX phases was totally decomposed at 985°C.The Addition of MAX enhanced the Al-matrix and Ni-matrix.

Keywords: MAX phase, microstructures, composites, hardness, SEM

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Authors: Renata Masarova, Bohuslava Juhasova, Martin Juhas, Zuzana Sutova

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In dynamic system theory a mathematical model is often used to describe their properties. In order to find a transfer matrix of a dynamic system we need to calculate an inverse matrix. The paper contains the fusion of the classical theory and the procedures used in the theory of automated control for calculating the inverse matrix. The final part of the paper models the given problem by the Matlab.

Keywords: dynamic system, transfer matrix, inverse matrix, modeling

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Authors: Behzad Hajalilou, Nasim Hajalilou, Saeid Ansari

Abstract:

The study area is located in East Azerbaijan province, north of Ahar city, and 1/100000 geological map of Varzgan. This region is located in the middle of Iran zone. Ali Javad Porphyry copper-gold ore deposit has been created in a magmatic complex containing intrusive masses, combining Granodiorite and quartz Monzonite that penetrates into the Eocene volcanic aggregate. The most important mineralization includes primary oxides minerals (magnetite), sulfide (pyrite, chalcopyrite, Molybdenite, Bornite, Chalcocite, Covollite), secondary oxide or hydroxide minerals (hematite, goethite, limonite), and carbonate (malachite and Azurite). The mineralization forms into the vein-veinlets and scattered system. The alterations observed in the region include intermediate Argillic, advanced Argillic, Phyllic, silica, Propylitic, chlorite and Potassic. The 3D model of mineralization of the Alijavad is provided by Data DATAMINE software and based on the study of 700 polished sections of 32 drilled boreholes in the region. This model is completely compatible with the model provided by Lowell and Gilbert for the mineralization of porphyry copper deposits of quartz Monzonite type. The estimated cumulative residual value of copper for Ali Javad deposit is 81.5 million tons with 0.75 percent of copper, and for gold is 8.37 million tons with 1.8 ppm.

Keywords: porphyry copper, mineralization, Ali Javad, modeling, reserve estimation

Procedia PDF Downloads 178

Authors: Mohamed A. Mohamed-Ali, Jannis Simos, Khalid M. Kheiralla

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The Ingessana hills in the southern Blue Nile of Sudan are part of the southern sector of the NE-SW trending ophiolithic belt of the Arab-Nubian Shield with mid-neoproterozoic age. The rocks are mainly serpentinized and in parts highly silicified dunites especially towards the contact with the intruding Bau granite. A promising chromite mineralization zones in the area tend to be generally associated with NE-SW trending shear-zones. A detailed geophysical survey employing electrical resistivity tomography (ERT) at 34 lines were carried out over a zone of a known chromite mineralization to test feasibility of detecting and delineating the ore (if exist) and accordingly facilitate the positioning of exploratory drill holes. ERT sections were inverted with smooth constraints inversion code where the contacts between the granite and the ultramafics are showing up clearly. The continuity of mineralization along the contact is not well confirmed. However, the low-resistivity anomalies are probably recognized as potential chromite mineralization zones. These anomalies represent prime targets for further exploration by drilling, trenching or shallow pits. If the results of the drilling or excavations are positive, small open pit exploitations may produce important tonnages of chromite.

Keywords: chromite exploration, ERT, Ingessana Hills, inversion

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Authors: A. Y. Tsidulko, T. M. Pankova, E. V. Grigorieva

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High-grade gliomas are the most frequent and most aggressive brain tumors which are characterized by active invasion of tumor cells into the surrounding brain tissue, where the extracellular matrix (ECM) plays a crucial role. Disruption of ECM can be involved in anticancer drugs effectiveness, side-effects and also in tumor relapses. The anti-inflammatory agent dexamethasone is a common drug used during high-grade glioma treatment for alleviating cerebral edema. Although dexamethasone is widely used in the clinic, its effects on normal brain tissue ECM remain poorly investigated. It is known that proteoglycans (PGs) are a major component of the extracellular matrix in the central nervous system. In our work, we studied the effects of dexamethasone on the ECM proteoglycans (syndecan-1, glypican-1, perlecan, versican, brevican, NG2, decorin, biglican, lumican) using RT-PCR in the experimental animal model. It was shown that proteoglycans in rat brain have age-specific expression patterns. In early post-natal rat brain (8 days old rat pups) overall PGs expression was quite high and mainly expressed PGs were biglycan, decorin, and syndecan-1. The overall transcriptional activity of PGs in adult rat brain is 1.5-fold decreased compared to post-natal brain. The expression pattern was changed as well with biglycan, decorin, syndecan-1, glypican-1 and brevican becoming almost equally expressed. PGs expression patterns create a specific tissue microenvironment that differs in developing and adult brain. Dexamethasone regimen close to the one used in the clinic during high-grade glioma treatment significantly affects proteoglycans expression. It was shown that overall PGs transcription activity is 1.5-2-folds increased after dexamethasone treatment. The most up-regulated PGs were biglycan, decorin, and lumican. The PGs expression pattern in adult brain changed after treatment becoming quite close to the expression pattern in developing brain. It is known that microenvironment in developing tissues promotes cells proliferation while in adult tissues proliferation is usually suppressed. The changes occurring in the adult brain after dexamethasone treatment may lead to re-activation of cell proliferation due to signals from changed microenvironment. Taken together obtained data show that dexamethasone treatment significantly affects the normal brain ECM, creating the appropriate microenvironment for tumor cells proliferation and thus can reduce the effectiveness of anticancer treatment and promote tumor relapses. This work has been supported by a Russian Science Foundation (RSF Grant 16-15-10243)

Keywords: dexamthasone, extracellular matrix, glioma, proteoglycan

Procedia PDF Downloads 156

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Emily Schlebes, Christian Hundhausen, Jens W. Fischer

Abstract:

The glycosaminoglycan hyaluronan (HA) is a major component of the extracellular matrix, typically produced by fibroblasts of the connective tissue but also by immune cells. Here, we investigated the capacity of human peripheral blood CD8 T cells from healthy donors to produce HA and to express HA receptors as well as HA degrading enzymes. Further, we evaluated the effect of pharmacological HA inhibition on CD8 T cell function. Using immunocytochemistry together with quantitative PCR analysis, we found that HA synthesis is rapidly induced upon antibody-induced T cell receptor (TCR) activation and almost exclusively mediated by HA synthase 3 (HAS3). TCR activation also resulted in the upregulation of HA receptors CD44, hyaluronan-mediated motility receptor (HMMR), and layilin (LAYN), although kinetics and strength of expression varied greatly between subjects. The HA-degrading enzymes HYAL1 and HYAL2 were detected at low levels and induced by cell activation in some individuals. Interestingly, expression of HAS3, HA receptors, and hyaluronidases were modulated by the proinflammatory cytokines IL-6 and IL-1bβ in most subjects. To assess the functional role of HA in CD8 T cells, we performed carboxyfluorescein succinimidyl ester (CFSE) based proliferation assays and cytokine analysis in the presence of the HA inhibitor 4- Methylumbelliferone (4-MU). Despite significant inter-individual variation with regard to the effective dose, 4-MU resulted in the inhibition of CD8 T cell proliferation and reduced release of TNF-α and IFN-γ. Collectively, these data demonstrate that human CD8 T cells respond to TCR stimulation with a synthesis of HA and expression of HA-related genes. They further suggest that HA inhibition may be helpful in interfering with pathogenic T cell activation in human disease.

Keywords: CD8 T cells, extracellular matrix, hyaluronan, hyaluronan synthase 3

Procedia PDF Downloads 62

Authors: K. M. Kheiralla, M. A. Ali, M. Y. Abdelgalil, N. E. Mohamed, G. Boutsis

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The present study focus on integrated geophysical surveys carried out in the mineralization zone in Erkowit region, Eastern Sudan to determine the extensions of the potential ore deposits on the topographically high hilly area and under the cover of alluvium along the nearby wadi and to locate other occurrences if any. The magnetic method (MAG) and the electrical resistivity tomography (ERT) were employed for the survey. Eleven traverses were aligned approximately at right angles to the general strike of the rock formations. The disseminated sulfides are located on the alteration shear zone which is composed of granitic and dioritic highly ferruginated rock occupying the southwestern and central parts of the area, this was confirmed using thin and polished sections mineralogical analysis. The magnetic data indicates low magnetic values for wadi sedimentary deposits in its southern part of the area, and high anomalies which are suspected as gossans due to magnetite formed during wall rock alteration consequent to mineralization. The significant ERT images define low resistivity zone as traced as sheared zones which may associated with the main loci of ore deposition. The study designates that correlation of magnetic and ERT anomalies with lithology are extremely useful in mineral exploration due to variations in some specific physical properties of rocks.

Keywords: ERT, magnetic, mineralization, Red Sea, Sudan

Procedia PDF Downloads 353