Search results for: macrophage activation

Authors: Ihab Elaff

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Simulating the heart’s electrical stimulation is essential in modeling and evaluating the electrophysiology behavior of the heart. For achieving that, there are two structures in concern: the ventricles’ Myocardium, and the ventricles’ Conduction Network. Ventricles’ Myocardium has been modeled as anisotropic material from Diffusion Tensor Imaging (DTI) scan, and the Conduction Network has been extracted from DTI as a case-based structure based on the biological properties of the heart tissues and the working methodology of the Magnetic Resonance Imaging (MRI) scanner. Results of the produced activation were much similar to real measurements of the reference model that was presented in the literature.

Keywords: diffusion tensor, DTI, heart, conduction network, excitation propagation

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Authors: Mariam A. Abu-Alim

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The purpose of this study was to validate muscle activation amplitudes of two field base dynamic balance tests that are used as strengthen and motor control exercises too in the activation of selected hip and knee stabilizer muscles. Methods: Eighteen college-age females students (21±2 years; 65.6± 8.7 kg; 169.7±8.1 cm) who participated at least for 30 minutes in physical activity most days of the week volunteered. The wireless BIOPAC (MP150, BIOPAC System. Inc, California, USA) surface electromyography system was used to validate the activation of the Gluteus Medius and the Adductor Magnus of hip stabilizer muscles; and the Hamstrings, Quadriceps, and the Gastrocnemius of the knee stabilizer muscles. Surface electrodes (EL 503, BIOPAC, System. Inc) connected to dual wireless EMG BioNormadix Transmitters were place on selected muscles of participants dominate side. Manual muscle testing was performed to obtain the maximal voluntary isometric contraction (MVIC) in which all collected muscle activity data during the three reaching direction: anterior, posteromedial, posterolateral of the Star Excursion Balance Test (SEBT) and the Y-balance Test (YBT) data could be normalized. All participants performed three trials for each reaching direction of the SEBT and the YBT. The domanial leg trial for each participant was selected for analysis which was also the standing leg. Results: the selected hip stabilizer muscles (Gluteus Medius, Adductor Magnus) were both greater than 100%MVIC during the performance of the SEBT and in all three directions. Whereas, selected knee stabilizer muscles had greater activation 0f 100% MVIC and were significantly more activated during the performance of the YBT test in all three reaching directions. The results showed that the posterolateral and the postmedial reaching directions for both dynamic balance tests had greater activation levels and greater than 200%MVIC for all tested muscles expect of the hamstrings. Conclusion: the results of this study showed that the SEBT and the YBT had validated high levels of muscular activity for the hip and the knee stabilizer muscles; which can be used to represent the improvement, strength, control and the decreasing in the injury levels. Since these selected hip and knee stabilizer muscles, represent 35% of all athletic injuries depending on the type of sport.

Keywords: dynamic balance tests, electromyography, hip stabilizer muscles, nee stabilizer muscles

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Authors: Suresh Vanguri, Suresh Palla, K. V. Kalyani, S. K. Chaturvedi, B. N. Mohapatra

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Enhancing the fly ash utilization in the manufacture of cement is identified as one of the key areas to mitigate the Green House Gas emissions from the cement industry. Though increasing the fly ash content in cement has economic and environmental benefits, it results in a decrease in the compressive strength values, particularly at early ages. Quality of clinker and fly ash were identified as predominant factors that govern the extent of absorption of fly ash in the manufacturing of cement. This paper presents systematic investigations on the effect of clinker and fly ash quality on the properties of resultant cement. Since mechanical activation alters the physicochemical properties such as particle size distribution, surface area, phase morphology, understanding the variation of these properties with activation is required for its applications. The effect of mechanical activation on fly ash surface area, specific gravity, flow properties, lime reactivity, comparative compressive strength (CCS), reactive silica and mineralogical properties were also studied. The fineness of fly ash was determined by Blaine’s method, specific gravity, lime reactivity, CCS were determined as per the method IS 1727-1967. The phase composition of fly ash was studied using the X-ray Diffraction technique. The changes in the microstructure and morphology with activation were examined using the scanning electron microscope. The studies presented in this paper also include evaluation of Portland Pozzolana Cement (PPC), prepared using high volume fly ash. Studies are being carried out using clinker from cement plants located in different regions/clusters in India. Blends of PPC containing higher contents of activated fly ash have been prepared and investigated for their chemical and physical properties, as per Indian Standard procedures. Changes in the microstructure of fly ash with activation and mechanical properties of resultant cement containing high volumes of fly ash indicated the significance of optimization of the quality of clinker and fly ash fineness for better techno-economical benefits.

Keywords: flow properties, fly ash enhancement, lime reactivity, microstructure, mineralogy

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

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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

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Authors: Alhulw H. Alshammari, Ahmed Iraqi, S. A. Saad, T. A. Taha

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In this work, we present for the first time the study of Raman spectra and dielectric relaxation of polyester polymer-CoFe₂O₄ (5.0, 10.0, 15.0, and 20.0 wt%) nanocomposites. Raman spectroscopy was applied as a sensitive structural identification technique to characterize the polyester-CoFe₂O₄ nanocomposites. The images of AFM confirmed the uniform distribution of CoFe₂O₄ inside the polymer matrix. Dielectric relaxation was employed as an important analytical technique to obtain information about the ability of the polymer nanocomposites to store and filter electrical signals. The dielectric relaxation analyses were carried out on the polyester-CoFe₂O₄ nanocomposites at different temperatures. An increase in dielectric constant ε₁ was observed for all samples with increasing temperatures due to the alignment of the electric dipoles with the applied electric field. In contrast, ε₁ decreased with increasing frequency. This is attributed to the difficulty for the electric dipoles to follow the electric field. The α relaxation peak that appeared at a high frequency shifted to higher frequencies when increasing the temperature. The activation energies for Maxwell-Wagner Sillar (MWS) changed from 0.84 to 1.01 eV, while the activation energies for α relaxations were 0.54 – 0.94 eV. The conduction mechanism for the polyester- CoFe₂O₄ nanocomposites followed the correlated barrier hopping (CBH) model.

Keywords: AC conductivity, activation energy, dielectric permittivity, polyester nanocomposites

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Authors: Nagarjuna Remalli, Robert Brandt

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Down-sizing of combustion engines in automobiles are prevailed owing to required increase in efficiency. This leads to a stress increment on valve springs, which affects their intended function due to an increase in relaxation. High strength martensitic steels are used for valve spring applications. Recent investigations unveiled that low temperature creep (LTC) in martensitic steels obey a logarithmic creep law. The exhaustion theory links the logarithmic creep behavior to an activation energy which is characteristic for any given time during creep. This activation energy increases with creep strain due to barriers of low activation energies exhausted during creep. The assumption of the exhaustion theory is that the material is inhomogeneous in microscopic scale. According to these assumptions it is anticipated that small obstacles (e. g. ε–carbides) having a wide range of size distribution are non-uniformly distributed in the materials. X-ray diffraction studies revealed the presence of ε–carbides in high strength martensitic steels. In this study, high strength martensitic steels that are crept in the temperature range of 75 – 150 °C were investigated with the aid of a transmission electron microscope for the evidence of an inhomogeneous distribution of obstacles having different size to examine the validation of exhaustion theory.

Keywords: creep mechanisms, exhaustion theory, low temperature creep, martensitic steels

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Authors: Asiya Mahtab, Sushama Talegaonkar

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The research is aimed at developing targeted lipid-based nanocarrier systems of chondroitin sulfate (CS) to deliver an antirheumatic drug to the inflammatory site in arthritic paw. Lipid-based nanoparticle (TEF-lipo) was prepared by using a thin-film hydration method. The coating of prepared drug-loaded nanoparticles was done by the ionic interaction mechanism. TEF-lipo and CS-coated lipid nanoparticle (CS-lipo) were characterized for mean droplet size, zeta potential, and surface morphology. TEF-lipo and CS-lipo were further subjected to in vitro cell line studies on RAW 264.7 murine macrophage, U937, and MG 63 cell lines. The pharmacodynamic study was performed to establish the effectiveness of the prepared lipid-based conventional and targeted nanoparticles in comparison to pure drugs. Droplet size and zeta potential of TEF-lipo were found to be 128. 92 ± 5.42 nm and +12.6 ± 1.2 mV. It was observed that after the coating of TEF-lipo with CS, particle size increased to 155.6± 2.12 nm and zeta potential changed to -10.2± 1.4mV. Transmission electron microscopic analysis revealed that the nanovesicles were uniformly dispersed and detached from each other. Formulations followed sustained release pattern up to 24 h. Results of cell line studies ind icated that CS-lipo formulation showed the highest cytotoxic potential, thereby proving its enhanced ability to kill the RAW 264.7 murine macrophage and U937 cells when compared with other formulations. It is clear from our in vivo pharmacodynamic results that targeted nanocarriers had a higher inhibitory effect on arthritis progression than nontargeted nanocarriers or free drugs. Results demonstrate that this approach will provide effective treatment for rheumatoid arthritis, and CS served as a potential prophylactic against the advancement of cartilage degeneration.

Keywords: adjuvant induced arthritis, chondroitin sulfate, rheumatoid arthritis, teriflunomide

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Authors: Mohammadjavad Sotoudeheian, Navid Farahmandian

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Kawasaki disease (KD) is the primary cause of acquired pediatric heart disease as an acute vasculitis. In children with prolonged fever, rash, and inflammation of the mucosa KD must be considered as a clinical diagnosis. There is a persuasive suggestion of immune-mediated damage as the pathophysiologic cascade of KD. For example, the invasion of cytotoxic T-cells supports a viral etiology and the inflammasome of the innate immune system is a critical component in the vasculitis formation in KD. Animal models of KD propose the cytokine profiles, such as increased IL-1 and GM-CSF, which cause vascular damage. CRP and IFN-γ elevated expression and the upregulation of IL-6, and IL-10 production are also described in previous studies. Untreated KD is a critical risk factor for coronary artery diseases and myocardial infarction. Vascular damage may encompass amplified T-cell activity. SMAD3 is an essential molecule in down-regulating T-cells and increasing expression of FoxP3. It has a critical effect in the differentiation of regulatory T-cells. The discrepancy of regulatory T-cells and pro-inflammatory Th17 has been studied in acute coronary syndrome during KD. However in the coronary artery damaged lymphocytes and IgA plasma cells are seen at the lesion locations, the major immune cells in the coronary lesions are monocytes/macrophages and neutrophils. These cells secrete TNF-α, and activates matrix metalloprotease (MMP)-9, reducing the integrity of vessels and prompting patients to arise aneurysm. MMPs can break down the components of the extracellular matrix and assist immune cell movement. IVIG as an effective form of treatment clarified the role of the immune system, which may target pathogenic antigens and regulate cytokine production. Several reports have revealed that in the coronary arteries, high expression of MMP-9 in monocyte/macrophage results in pathologic cascades. Curcumin is a potent antioxidant and anti-inflammatory molecule. Curcumin decreases the production of reactive oxygen and nitrogen species and inhibits transcription factors like AP-1 and NF-κB. Curcumin also contains the characteristics of inhibitory effects on MMPs, especially MMP-9. The upregulation of MMP-9 is an important cellular response. Curcumin treatment caused a reverse effect and down-regulates MMP-9 gene expression which may fund the anti-inflammatory effect. Curcumin inhibits MMP-9 expression via PKC and AMPK-dependent pathways in Human monocytes cells. Elevated expression and activity of MMP-9 are correlated with advanced vascular lesions. AMPK controls lipid metabolism and oxidation, and protein synthesis. AMPK is also necessary for the MMP-9 activity and THP-1 cell adhesion to endothelial cells. Curcumin was shown to inhibit the activation of AMPKα. Compound C (AMPK inhibitor) inhibits MMP-9 expression level. Therefore, through inactivating AMPKs and PKC, curcumin decreases the MMP-9 level, which results in inhibiting monocyte/macrophage differentiation. Compound C also suppress the phosphorylation of three major classes of MAP kinase signaling, suggesting that curcumin may suppress MMP-9 level by inactivation of MAPK pathways. MAPK cascades are activated to induce the expression of MMP-9. Curcumin inhibits MAPKs phosphorylation, which contributes to the down-regulation of MMP-9. This study demonstrated that the potential inhibitory properties of curcumin over MMP-9 lead to a therapeutic strategy to reduce the risk of coronary artery involvement during KD.

Keywords: MMP-9, coronary artery aneurysm, Kawasaki’s disease, curcumin, AMPK, immune system, NF-κB, MAPK

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Authors: Seyedahmad Hosseini, Mohammadjavad Sotoudeheian

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Introduction: Allergic asthma is a heterogeneous immuno-inflammatory disease based on Th-2-mediated inflammation. Histopathologic abnormalities of the airways characteristic of asthma include epithelial damage and subepithelial collagen deposition. Objectives: Human bronchial epithelial cell genome expression of TNF‑α, IL‑6, ICAM‑1, VCAM‑1, nuclear factor (NF)‑κB signaling pathways up-regulate during inflammatory cascades. Moreover, immunofluorescence assays confirmed the nuclear translocation of NF‑κB p65 during inflammatory responses. An absolute LDH leakage assays suggestedLPS-inducedcells injury, and the associated mechanisms are co-incident events. LPS-induced phosphorylation of ERKand JNK causes inflammation in epithelial cells through inhibition of ERK and JNK activation and NF-κB signaling pathway. Furthermore, the inhibition of NF-κB mRNA expression and the nuclear translocation of NF-κB lead to anti-inflammatory events. Likewise, activation of SUMF2 which inhibits IL-13 and reduces Th2-cytokines, NF-κB, and IgE levels to ameliorate asthma. On the other hand, TNFα-induced mucus production reduced NF-κB activation through inhibition of the activation status of Rac1 and IκBα phosphorylation. In addition, bradykinin B2 receptor (B2R), which mediates airway remodeling, regulates through NF-κB. Bronchial B2R expression is constitutively elevated in allergic asthma. In addition, certain NF-κB -dependent chemokines function to recruit eosinophils in the airway. Besides, bromodomain containing 4 (BRD4) plays a significant role in mediating innate immune response in human small airway epithelial cells as well as transglutaminase 2 (TG2), which is detectable in saliva. So, the guanine nucleotide-binding regulatory protein α-subunit, Gα16, expresses a κB-driven luciferase reporter. This response was accompanied by phosphorylation of IκBα. Furthermore, expression of Gα16 in saliva markedly enhanced TNF-α-induced κB reporter activity. Methods: The applied method to form NF-κB activation is the electromobility shift assay (EMSA). Also, B2R-BRD4-TG2 complex detection by immunoassay method within saliva with EMSA of NF-κB activation may be a novel biomarker for asthma diagnosis and follow up. Conclusion: This concept introduces NF-κB signaling pathway as potential asthma biomarkers and promising targets for the development of new therapeutic strategies against asthma.

Keywords: NF-κB, asthma, saliva, T-helper

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Authors: Chaoran Liu

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Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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Authors: Mayada Mazher, Ahmed Moustafa, Ahmed Abdellatif

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Background: Autophagy is a eukaryotic, highly conserved catabolic process implicated in many pathophysiologies such as wound healing. Autophagy-associated genes serve as a scaffolding platform for signal transduction of macrophage polarization during the inflammatory phase of wound healing and tissue repair process. In the current study, we report a model for the interplay between autophagy-associated genes and macrophages polarization associated genes. Methods: In silico analysis was performed on 249 autophagy-related genes retrieved from the public autophagy database and gene expression data retrieved from Gene Expression Omnibus (GEO); GSE81922 and GSE69607 microarray data macrophages polarization 199 DEGS. An integrated protein-protein interaction network was constructed for autophagy and macrophage gene sets. The gene sets were then used for GO terms pathway enrichment analysis. Common transcription factors for autophagy and macrophages' polarization were identified. Finally, microRNAs enriched in both autophagy and macrophages were predicated. Results: In silico prediction of common transcription factors in DEGs macrophages and autophagy gene sets revealed a new role for the transcription factors, HOMEZ, GABPA, ELK1 and REL, that commonly regulate macrophages associated genes: IL6,IL1M, IL1B, NOS1, SOC3 and autophagy-related genes: Atg12, Rictor, Rb1cc1, Gaparab1, Atg16l1. Conclusions: Autophagy and macrophages' polarization are interdependent cellular processes, and both autophagy-related proteins and macrophages' polarization related proteins coordinate in tissue remodelling via transcription factors and microRNAs regulatory network. The current work highlights a potential new role for transcription factors HOMEZ, GABPA, ELK1 and REL in wound healing.

Keywords: autophagy related proteins, integrated network analysis, macrophages polarization M1 and M2, tissue remodelling

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Authors: Miao Zhang, Limin Liu, Feng Zhi, Panpan Niu, Mengya Yang, Xuemei Zhu, Ying Diao, Jun Wang, Ying Zhao

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Background and Aims: Clinical studies have demonstrated that serum semicarbazide-sensitive amine oxidase (SSAO) activities positively correlate with the progression of atherosclerosis. The aim of the present study is to investigate the effect of SSAO inactivation on the development of atherosclerosis. Methods: Female LDLr knockout (KO) mice were given the Western-type diet for 6 and 9 weeks to induce the formation of early and advanced lesions, and semicarbazide (SCZ, 0.125%) was added into the drinking water to inactivate SSAO in vivo. Results: Despite no impact on plasma total cholesterol levels, abrogation of SSAO by SCZ not only resulted in the enlargement of both early (1.5-fold, p=0.0043) and advanced (1.8-fold, p=0.0013) atherosclerotic lesions, but also led to reduced/increased lesion contents of macrophages/smooth muscle cells (SMCs) (macrophage: ~0.74-fold, p=0.0002(early)/0.0016(advanced); SMC: ~1.55-fold, p=0.0003(early) /0.0001(advanced)), respectively. Moreover, SSAO inactivation inhibited the migration of circulating monocytes into peripheral tissues and reduced the amount of circulating Ly6Chigh monocytes (0.7-fold, p=0.0001), which may account for the reduced macrophage content in lesions. In contrast, the increased number of SMCs in lesions of SCZ-treated mice is attributed to an augmented synthetic vascular SMC phenotype switch as evidenced by the increased proliferation of SMCs and accumulation of collagens in vivo. Conclusion: SSAO inactivation by SCZ promotes the phenotypic switch of SMCs and the development of atherosclerosis. The enzymatic activity of SSAO may thus represent a potential target in the prevention and/or treatment of atherosclerosis.

Keywords: atherosclerosis, phenotype switch of smooth muscle cells, SSAO/VAP-1, semicarbazide

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Authors: Jay J. Vora, Vishvesh J. Badheka

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This work attempts to investigate the effect of oxide fluxes on 6mm thick Reduced Activation ferritic/martensitic steels (RAFM) during Activated TIG (A-TIG) welding. Six different fluxes Al₂O₃, Co₃O₄, CuO, HgO, MoO₃, and NiO were mixed with methanol for conversion into paste and bead-on-plate experiments were then carried out. This study, systematically investigates the influence of oxide-based flux powder and carrier solvent composition on the weld bead shape, geometric shape of weld bead and dominant depth enhancing mechanism in tungsten inert gas (TIG) welding of reduced activation ferritic/martensitic (RAFM) steel. It was inferred from the study that flux Co₃O₄ and MoO₃ imparted full and secure (more than 6mm) penetration with methanol owing to dual mechanism of reversed Marangoni and arc construction. The use of methanol imparted good spreadabilty and coverability and ultimately higher peak temperatures were observed with its use owing to stronger depth enhancing mechanisms than use of acetone with same oxide fluxes and welding conditions.

Keywords: A-TIG, flux, oxides, penetration, RAFM, temperature, welding

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Authors: Jianli Dong, Fangling Xu, Gengming Huang

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Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.

Keywords: CD4 cell, HERV-K, HIV-1, melanoma

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Authors: Kefale W. Yizengaw, Delele Worku Ayele, Jyh-Chiang Jiang

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The catalytic co-conversion of CO₂ and CH₄ to value-added compounds has become one of the promising approaches to addressing global climate change by having valuable fossil fuels. Thedirect co-conversion of CO₂ and CH₄ to value-added compounds is attractive but tremendously challenging because of both molecules' thermodynamic stability and kinetic inertness. In the present study, a single iridium atom doped and a single oxygen atom defect hematite (110)surface model catalyst, which can comprehend direct C–O coupling based on simultaneous activation of CO2 and CH4 was studied using density functional theory plus U (DFT + U)calculations. The presence of dual active sites on the Ir/Fe₂O₃(110)-OV surface catalyst enablesCO₂ activation on the Ir site and CH₄ activation at the defect site. The electron analysis for the theco-adsorption of CO₂ and CH₄ deals with the electron redistribution on the surface and clearly shows the synergistic effect for simultaneous CO₂ and CH₄ activation on Ir/α- Fe₂O₃(110)-OVsurface. The microkinetic analysis shows that the dissociation of CH4 to CH3 * and H* plays an excellent role in the C–O coupling. The coverage analysis for the intermediate products of the microkinetic simulation results indicates that C–O coupling is the reaction limiting step. Finally, after the CH₃O* intermediate product species is produced, the radical hydrogen species spontaneously diffuse to the CH3O* intermediate product to form methanol at around 490 [K]. The present work provides mechanistic and kinetic insights into the direct C–O coupling of CO₂and CH₄, which could help design more-efficient catalysts.

Keywords: co-conversion, C–O coupling, doping, oxygen vacancy, microkinetic

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Authors: Virgil Miranda, Gissele Mosqueda, Pablo Delgado, Yimesker Yihun

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Muscular fatigue affects the muscle activation that is needed for producing the desired clinical outcome. Integrating optimal muscle relaxation periods into a variety of health care rehabilitation protocols is important to maximize the efficiency of the therapy. In this study, four muscle relaxation periods (30, 60, 90, and 120 seconds) and their effectiveness in producing consistent muscle activation of the muscle biceps brachii between sets of elbow flexion and extension task was investigated among a sample of 10 subjects with no disabilities. The same resting periods were then utilized in a controlled exoskeleton-based exercise for a sample size of 5 subjects and have shown similar results. On average, the muscle activity of the biceps brachii decreased by 0.3% when rested for 30 seconds, and it increased by 1.25%, 0.76%, and 0.82% when using muscle relaxation periods of 60, 90, and 120 seconds, respectively. The preliminary results suggest that a muscle relaxation period of about 60 seconds is needed for optimal continuous muscle activation within rehabilitation regimens. Robot-based rehabilitation is good to produce repetitive tasks with the right intensity, and knowing the optimal resting period will make the automation more effective.

Keywords: rest intervals, muscle biceps brachii, robot rehabilitation, muscle fatigue

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Authors: Ronald Croce, Timothy Quinn, John Miller

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Incomplete activation, or activation failure, of motor units during maximal voluntary contractions is often referred to as muscle inhibition (MI), and is defined as the inability of the central nervous system to maximally drive a muscle during a voluntary contraction. The purpose of the present study was to assess the interrelationship amongst peak torque (PT), muscle inhibition (MI; incomplete activation of motor units), and voluntary muscle activation (VMA) of the quadriceps’ muscle group as a function of knee angle and muscle length during maximal voluntary isometric contractions (MVICs). Nine young adult males (mean + standard deviation: age: 21.58 + 1.30 years; height: 180.07 + 4.99 cm; weight: 89.07 + 7.55 kg) performed MVICs in random order with the knee at 15, 55, and 95° flexion. MI was assessed using the interpolated twitch technique and was estimated by the amount of additional knee extensor PT evoked by the superimposed twitch during MVICs. Voluntary muscle activation was estimated by root mean square amplitude electromyography (EMGrms) and mechanomyography (MMGrms) of agonist (vastus medialis [VM], vastus lateralis [VL], and rectus femoris [RF]) and antagonist (biceps femoris ([BF]) muscles during MVICs. Data were analyzed using separate repeated measures analysis of variance. Results revealed a strong dependency of quadriceps’ PT (p < 0.001), MI (p < 0.001) and MA (p < 0.01) on knee joint position: PT was smallest at the most shortened muscle position (15°) and greatest at mid-position (55°); MI and MA were smallest at the most shortened muscle position (15°) and greatest at the most lengthened position (95°), with the RF showing the greatest change in MA. It is hypothesized that the ability to more fully activate the quadriceps at short compared to longer muscle lengths (96% contracted at 15°; 91% at 55°; 90% at 95°) might partly compensate for the unfavorable force-length mechanics at the more extended position and consequent declines in VMA (decreases in EMGrms and MMGrms muscle amplitude during MVICs) and force production (PT = 111-Nm at 15°, 217-NM at 55°, 199-Nm at 95°). Biceps femoris EMG and MMG data showed no statistical differences (p = 0.11 and 0.12, respectively) at joint angles tested, although there were greater values at the extended position. Increased BF muscle amplitude at this position could be a mechanism by which anterior shear and tibial rotation induced by high quadriceps’ activity are countered. Measuring and understanding the degree to which one sees MI and VMA in the QF muscle has particular clinical relevance because different knee-joint disorders, such ligament injuries or osteoarthritis, increase levels of MI observed and markedly reduced the capability of full VMA.

Keywords: electromyography, interpolated twitch technique, mechanomyography, muscle activation, muscle inhibition

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Authors: Apoorva Vinod, Archana Mathur, Snehanshu Saha

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Though there exists a plethora of Activation Functions (AFs) used in single and multiple hidden layer Neural Networks (NN), their behavior always raised curiosity, whether used in combination or singly. The popular AFs –Sigmoid, ReLU, and Tanh–have performed prominently well for shallow and deep architectures. Most of the time, AFs are used singly in multi-layered NN, and, to the best of our knowledge, their performance is never studied and analyzed deeply when used in combination. In this manuscript, we experiment with multi-layered NN architecture (both on shallow and deep architectures; Convolutional NN and VGG16) and investigate how well the network responds to using two different AFs (Sigmoid-Tanh, Tanh-ReLU, ReLU-Sigmoid) used alternately against a traditional, single (Sigmoid-Sigmoid, Tanh-Tanh, ReLUReLU) combination. Our results show that using two different AFs, the network achieves better accuracy, substantially lower loss, and faster convergence on 4 computer vision (CV) and 15 Non-CV (NCV) datasets. When using different AFs, not only was the accuracy greater by 6-7%, but we also accomplished convergence twice as fast. We present a case study to investigate the probability of networks suffering vanishing and exploding gradients when using two different AFs. Additionally, we theoretically showed that a composition of two or more AFs satisfies Universal Approximation Theorem (UAT).

Keywords: activation function, universal approximation function, neural networks, convergence

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Authors: Ediati Budi Cahyono, Triana Hertiani, Nauval Arrazy Asawimanda, Wahyu Puji Pratomo

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Background: Doxorubicin is widely used as a chemotherapeutic drug despite having many side effects. It may cause macrophage dysfunction and decreasing proliferation of lymphocyte. Noni (Morinda citrifolia) fruit which has rich of polysaccharide content has potential as antitumor and immunostimulant effect. The isolation of polysaccharide from Noni fruit has been optimized according to four different methods based on macrophage and lymphocyte activities. We found the highest polysaccharide content from one of the four methods isolation. A method of polysaccharide isolation which has the highest immunostimulant effect was used for further observation as co-chemotherapy. The aim of the study: was to evaluate the isolated polysaccharide from the method of choice as co-chemotherapy of doxorubicin for the growth of Vero, He-La, and T47D cell lines in vitro. The method: in vitro growth assay of Vero, He-La, and T47D cell lines was done using MTT-reduction method, and apoptosis test was done by double staining method to evaluate the induction apoptotic effect of the combination. Every group was treated with doxorubicin and isolated polysaccharide from method of choice with 4 variances of concentrations (25 µg/ml, 50 µg/ml, 100 µg/ml and 200 µg/ml) a long with negative control (doxorubicin only) and normal control (without doxorubicin or polysaccharide administration). Results: The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin against He-La and T47D cell lines influenced the highest cytotoxic effect by suppressing cell viability comparing with doxorubicin only. The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin-induced apoptotic effect the He-La cell line comparing with doxorubicin only. The result of the study: it can be concluded that the combination of polysaccharide fraction and doxorubicin effect more selective toward He-La and T47D cell lines than to Vero cell line. It can be suggested isolated polysaccharide from the method of choice has co-chemotherapy activity against doxorubicin.

Keywords: polysaccharide, noni fruit, doxorubicin, cancer cell lines, vero cell line

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Authors: Rajkumar Ghosh

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The study examines the structural analysis of Chaura Thrust in Himachal Pradesh, India, focusing on the activation timing of Main Central Thrust (MCT) and South Tibetan Detachment System (STDS), mylonitised zones, and the characterization of box fold and its signature in the regional geology of Himachal Himalaya. The research aims to document the Higher Himalayan Out-of-Sequence Thrust (OOST) in Himachal Pradesh, which activated the MCTL and in between a zone south of MCTU. The study also documents the GBM-associated temperature range and the activation of Higher Himalayan Out-of-Sequence Thrust (OOST) in Himachal Pradesh. The findings contribute to understanding the structural analysis of Chaura Thrust and its signature in the regional geology of Himachal Himalaya. The study highlights the significance of microscopic studies in documenting mylonitized zones and identifying various types of crenulated schistosity. The study concludes that Chaura Thrust is not a blind thrust and details the field evidence for the OOST. The study characterizes the box fold and its signature in the regional geology of Himachal Himalaya. The study also documents the activation timing and ages of MCT, STDS, MBT, and MFT and identifies various types of crenulated schistosity under the microscope. The study also highlights the significance of microscopic studies in the structural analysis of Chaura Thrust. Finally, the study documents the activation of Higher Himalayan Out-of-Sequence Thrust (OOST) in Himachal Pradesh and the expectations for strain variation near the OOST.

Keywords: Chaura Thrust, Higher Himalaya, Jhakri Thrust, Main Central Thrust, Out-of-Sequence Thrust, Sarahan Thrust

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Authors: Yu-Chen Hu

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Gene editing by CRISPR and gene regulation by microRNA or CRISPR activation have dramatically changed the way to manipulate cellular gene expression and cell fate. In recent years, various gene editing and gene manipulation technologies have been applied to control stem cell differentiation to enhance tissue regeneration. This research will focus on how to develop CRISPR, CRISPR activation (CRISPRa), CRISPR inhibition (CRISPRi), as well as bi-directional CRISPR-AI gene regulation technologies to control cell differentiation and bone regeneration. Moreover, in this study, CRISPR/Cas13d-mediated RNA editng for miRNA editing and bone regeneration will be discussed.

Keywords: gene therapy, bone regeneration, stem cell, CRISPR, gene regulation

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Authors: Michael K. Adu, Boniface K. Alese, Olumide S. Ogunnusi

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This paper brings to fore the inherent advantages in application of mobile agents to procure software products rather than downloading software content on the Internet. It proposes a system whereby the products come on compact disk with mobile agent as deliverable. The client/user purchases a software product, but must connect to the remote server of the software developer before installation. The user provides an activation code that activates mobile agent which is part of the software product on compact disk. The validity of the activation code is checked on connection at the developer’s end to ascertain authenticity and prevent piracy. The system is implemented by downloading two different software products as compare with installing same products on compact disk with mobile agent’s application. Downloading software contents from developer’s database as in the traditional method requires a continuously open connection between the client and the developer’s end, a fixed network is not economically or technically feasible. Mobile agent after being dispatched into the network becomes independent of the creating process and can operate asynchronously and autonomously. It can reconnect later after completing its task and return for result delivery. Response Time and Network Load are very minimal with application of Mobile agent.

Keywords: software products, software developer, internet, activation code, mobile agent

Procedia PDF Downloads 278

Authors: A. P. Srivastava, D. Srivastava, D. J. Browne

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Relaxation dynamics of Zr44Cu40Al8Ag8 bulk metallic glass (BMG) has been probed using dynamic mechanical analyzer. The BMG sample was casted in the form of a plate of dimension 55 mm x 40 mm x 3 mm using tilt casting technique. X-ray diffraction and transmission electron microscope have been used for the microstructural characterization of as-cast BMG. For the mechanical spectroscopy study, samples in the form of a bar of size 55 mm X 2 mm X 3 mm were machined from the BMG plate. The mechanical spectroscopy was performed on dynamic mechanical analyzer (DMA) by 50 mm 3-point bending method in a nitrogen atmosphere. It was observed that two glass transition process were competing in supercooled liquid region around temperature 390°C and 430°C. The supercooled liquid state was completely characterized using DMA and differential scanning calorimeter (DSC). In addition to the main α-relaxation process, presence of β relaxation process around temperature 360°C; below the glass transition temperature was also observed. The β relaxation process could be described by Arrhenius law with the activation energy of 160 kJ/mole. The volume of the flow unit associated with this relaxation process has been estimated. The results from DMA study has been used to characterize the shear transformation zone in terms of activation volume and size. High fragility parameter value of 34 and higher activation volume indicates that this alloy could show good plasticity in supercooled liquid region. The possible mechanism for the relaxation processes has been discussed.

Keywords: DMA, glass transition, metallic glass, thermoplastic forming

Procedia PDF Downloads 272

Authors: Beth Taylor, Kojima Mituaki, Atsushi Senda, Koji Morishita, Yasuhiro Otomo

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Intestinal ischaemia-reperfusion injury drives systemic inflammation and organ failure following trauma/haemorrhagic shock (T/HS), through the release of pro-inflammatory mediators into the mesenteric lymph (ML). However, changes in the biological function of ML are not fully understood, and therefore, a specific model of intestinal ischaemia-reperfusion injury is required to obtain ML for the study of its biological function upon inflammatory cells. ML obtained from a model of intestinal ischaemia-reperfusion injury was used to assess biological function upon inflammatory cells and investigate changes in the biological function of individual ML components. An additional model was used to determine the effect of vagal nerve stimulation (VNS) upon biological function. Rat ML was obtained by mesenteric lymphatic duct cannulation before and after occlusion of the superior mesenteric artery (SMAO). ML was incubated with human polymorphonuclear neutrophils (PMNs), monocytes and lymphocytes, and the biological function of these cells was assessed. ML was then separated into supernatant, exosome and micro-vesicle components, and biological activity was compared in monocytes. A model with an additional VNS phase was developed, in which the right cervical vagal nerve was exposed and stimulated, and ML collected for comparison of biological function with the conventional model. The biological function of ML was altered by intestinal ischaemia-reperfusion injury, increasing PMN activation, monocyte activation, and lymphocyte apoptosis. Increased monocyte activation was only induced by the exosome component of ML, with no significant changes induced by the supernatant or micro-vesicle components. VNS partially attenuated monocyte activation, but no attenuation of PMN activation was observed. Intestinal ischaemia-reperfusion injury induces changes in the biological function of ML upon both innate and adaptive inflammatory cells, supporting the role of intestinal ischaemia-reperfusion injury in driving systemic inflammation following T/HS. The exosome component of ML appears to be critical to the transport of pro-inflammatory mediators in ML. VNS partially attenuates changes in innate inflammatory cell biological activity observed, presenting possibilities for future novel treatment development in multiple organ failure patients.

Keywords: exosomes, inflammation, intestinal ischaemia, mesenteric lymph, vagal stimulation

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Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Elham Javanroudi

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Introduction: Tuberculosis (TB) is a known disease with hidden features caused by Mycobacterium tuberculosis (MTB). This disease, which is one of the 10 deadliest in the world, has caused millions of deaths in recent decades. Furthermore, TB is responsible for infecting about 30% population of world. Like any infection, TB can activate the immune system by locating and colonization in the human body, especially in the alveoli. TB is granulomatosis, so MTB can absorb the host’s immune cells and other cells to form granuloma. Method: Different databases (e.g., PubMed) were recruited to prepare this paper and fulfill our goals to search and find effective papers and investigations. Results: Immune response to MTB is related to T cell killers and contains CD1, CD4, and CD8 T lymphocytes. CD1 lymphocytes can recognize glycolipids, which highly exist in the Mycobacterial fatty cell wall. CD4 lymphocytes and macrophages form granuloma, and it is the main line of immune response to Mycobacteria. On the other hand, CD8 cells have cytolytic function for directly killing MTB by secretion of granulysin. Other functions and secretion to the deal are interleukin-12 (IL-12) by induction of expression interferon-γ (INF-γ) for macrophages activation and creating a granuloma, and tumor necrosis factor (TNF) by promoting macrophage phagolysosomal fusion. Conclusion: Immune cells in battle with MTB are macrophages, dendritic cells (DCs), neutrophils, and natural killer (NK) cells. These immune cells can recognize the Mycobacterium by various receptors, including Toll-like receptors (TLRs), Nod-like receptors (NLRs), and C-type lectin receptors (CLRs) located in the cell surface. In human alveoli exist about 50 dendritic macrophages, which have close communication with other immune cells in the circulating system and epithelial cells to deal with Mycobacteria. Against immune cells, MTB handles some factors (e.g., cordfactor, O-Ag, lipoarabinomannan, sulfatides, and adenylate cyclase) and practical functions (e.g., inhibition of macrophages).

Keywords: mycobacterium tuberculosis, immune responses, immunological mechanisms, respiratory tuberculosis

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Authors: Michael R. Shurin, Galina Shurin, Vladimir A. Kirichenko

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Background. Visibility of hepatic malignancies is poor on non-contrast imaging for daily verification of liver malignancies prior to radiation therapy on MRI-guided Linear Accelerators (MR-Linac). Ferumoxytol® (Feraheme, AMAG Pharmaceuticals, Waltham, MA) is a SPION agent that is increasingly utilized off-label as hepatic MRI contrast. This agent has the advantage of providing a functional assessment of the liver based upon its uptake by hepatic Kupffer cells proportionate to vascular perfusion, resulting in strong T1, T2 and T2* relaxation effects and enhanced contrast of malignant tumors, which lack Kupffer cells. The latter characteristic has been recently utilized for MRI-guided radiotherapy planning with precision targeting of liver malignancies. However potential radiotoxicity of SPION has never been addressed for its safe use as an MRI-contrast agent during liver radiotherapy on MRI-Linac. This study defines the radiomodulating properties of SPIONs in vitro on human monocyte and macrophage cell lines exposed to 60Go gamma-rays within clinical radiotherapy dose range. Methods. Human monocyte and macrophages cell line in cultures were loaded with a clinically relevant concentration of Ferumoxytol (30µg/ml) for 2 and 24 h and irradiated to 3Gy, 5Gy and 10Gy. Cells were washed and cultured for additional 24 and 48 h prior to assessing their phenotypic activation by flow cytometry and function, including viability (Annexin V/PI assay), proliferation (MTT assay) and cytokine expression (Luminex assay). Results. Our results reveled that SPION affected both human monocytes and macrophages in vitro. Specifically, iron oxide nanoparticles decreased radiation-induced apoptosis and prevented radiation-induced inhibition of human monocyte proliferative activity. Furthermore, Ferumoxytol protected monocytes from radiation-induced modulation of phenotype. For instance, while irradiation decreased polarization of monocytes to CD11b+CD14+ and CD11bnegCD14neg phenotype, Ferumoxytol prevented these effects. In macrophages, Ferumoxytol counteracted the ability of radiation to up-regulate cell polarization to CD11b+CD14+ phenotype and prevented radiation-induced down-regulation of expression of HLA-DR and CD86 molecules. Finally, Ferumoxytol uptake by human monocytes down-regulated expression of pro-inflammatory chemokines MIP-1α (Macrophage inflammatory protein 1α), MIP-1β (CCL4) and RANTES (CCL5). In macrophages, Ferumoxytol reversed the expression of IL-1RA, IL-8, IP-10 (CXCL10) and TNF-α, and up-regulates expression of MCP-1 (CCL2) and MIP-1α in irradiated macrophages. Conclusion. SPION agent Ferumoxytol increases resistance of human monocytes to radiation-induced cell death in vitro and supports anti-inflammatory phenotype of human macrophages under radiation. The effect is radiation dose-dependent and depends on the duration of Feraheme uptake. This study also finds strong evidence that SPIONs reversed the effect of radiation on the expression of pro-inflammatory cytokines involved in initiation and development of radiation-induced liver damage. Correlative translational work at our institution will directly assess the cyto-protective effects of Ferumoxytol on human Kupfer cells in vitro and ex vivo analysis of explanted liver specimens in a subset of patients receiving Feraheme-enhanced MRI-guided radiotherapy to the primary liver tumors as a bridge to liver transplant.

Keywords: superparamagnetic iron oxide nanoparticles, radioprotection, magnetic resonance imaging, liver

Procedia PDF Downloads 52

Authors: Lídia E. O. Cruz, Adriano P. C. Calvo, Renato J. Soares, Regiane A. Carvalho

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Individuals with non-specific chronic low back pain may present altered movement patterns during functional activities. However, muscle behavior before and after performing a functional task with different load conditions is not yet fully understood. The aim of this study is to analyze lumbar muscle activity before and after performing the functional task of picking up and placing an object on the ground (with and without load) in individuals with nonspecific chronic low back pain. 20 subjects with nonspecific chronic low back pain and 20 healthy subjects participated in this study. A surface electromyography was performed in the ilio-costal, longissimus and multifidus muscles to evaluate lumbar muscle activity before and after performing the functional task of picking up and placing an object on the ground, with and without load. The symptomatic participants had greater lumbar muscle activation compared to the asymptomatic group, more evident in performing the task without load, with statistically significant difference (p = 0,033) between groups for the right multifidus muscle. This study showed that individuals with nonspecific chronic low back pain have higher muscle activation before and after performing a functional task compared to healthy participants.

Keywords: chronic low back pain, functional task, lumbar muscles, muscle activity

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Authors: Hannele Laaksonen, Seija Nyqvist, Kari Nurmes

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Objectives: In the year 2011 the City of Vaasa started to develop know-how in the technology and the introduction of services for aging people in cooperation with the Polytechnic Novia University of Applied Sciences and VAMK, University of Applied Sciences. The project´s targets included: to help elderly people to maintain their ability to function, to provide them social and physical activities, to prevent their social exclusion, to decrease their feelings of loneliness and insecurity and to develop their technical know-how. Methods: The project was built based on open source code, tailor-made service system and user interface for the elderly living at home and their families, based on the users´ expectations and experiences of services. Activation-TV®-project vas carried out 1.4.2011-31.3.2014. A pilot group of eight elderly persons, who were living at home, were selected to the project. All necessary technical means as well as guidance and teaching equipment were provided to the pilot group. The students of University of Applied Sciences (VAMK, Novia) and employees of Center of Ageing were made all programs to the Activation-TV®. The project group were interviewed after and before intervention. The data were evaluated both qualitatively and quantitatively. Results: The built service includes a video library, a group room for interactive programs and a personal room for bilateral meetings and direct shipment. The program is bilingual and produced in both national languages. The Activation TV® reduced elderly peoples´ (n=8) feelings of emptiness, added mental well-being and quality of life with social contacts. Relatives felt, that they were able to get in to older peoples´ everyday life with Activation TV®. Discussion: The built application was tailored to the model that has not been developed elsewhere in Finland. This model can be copied from one server to another and thus transferred to other municipalities but the program requires its own personnel system management and maintenance as well as program production cooperation between the different actors. This service can be used for the elderly who are living at home without dementia.

Keywords: mental well-being, quality of life, elderly people, Finland

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Authors: Nafisa Komilova, Plamena Angelova, Andrey Abramov, Ulugbek Mirkhodjaev

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Parkinson’s disease (PD) is a progressive neurodegenerative disorder which is induced by the loss of dopaminergic neurons in the midbrain. The mechanism of neurodegeneration is associated with the aggregation of misfolded proteins, oxidative stress, and mitochondrial disfunction. Considering this, the process of removal of unwanted organelles or proteins by autophagy is vitally important in neurons, and activation of these processes could be protective in PD. Short-time acidification of cytosol can activate mitophagy and autophagy, and here we used sodium pyruvate and sodium lactate in human fibroblasts with PD mutations (Pink1, Pink1/Park2, α-syn triplication, A53T) to induce changes in intracellular pH. We have found that both lactate and pyruvate in millimolar concentrations can induce short-time acidification of cytosol in these cells. It induced activation of mitophagy and autophagy in control and PD fibroblasts and protected against cell death. Importantly, the application of lactate to acute brain slices of control and Pink1 knockout mice also induced a reduction of pH in neurons and astrocytes that increase the level of mitophagy. Thus, acidification of cytosol by compounds which play important role in cell metabolism also can activate mitophagy and autophagy and protect cells in the familial form of PD.

Keywords: Parkinson's disease, mutations, mitophagy, autophagy

Procedia PDF Downloads 166

Authors: Eduardo Kanagushiku Pereira, Frank Gregory Cavalcante da Silva, Barbara Soares Gonçalves, Ana Lúcia Bergamasco Galastri, Ronei Luciano Mamoni

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The inflammatory response initiates after the recognition of pathogens by receptors expressed by innate immune cells. Among these receptors, the NLRP3 was associated with the recognition of pathogenic fungi in experimental models. NLRP3 operates forming a multiproteic complex called inflammasome, which actives caspase-1, responsible for the production of the inflammatory cytokines IL-1beta and IL-18. In this study, we aimed to investigate the involvement of NLRP3 in the inflammatory response elicited in macrophages against Paracoccidioides brasiliensis (Pb), the etiologic agent of PCM. Macrophages were differentiated from THP-1 cells by treatment with phorbol-myristate-acetate. Following differentiation, macrophages were stimulated by Pb yeast cells for 24 hours, after previous treatment with specific NLRP3 (3,4-methylenedioxy-beta-nitrostyrene) and/or caspase-1 (VX-765) inhibitors, or specific inhibitors of pathways involved in NLRP3 activation such as: Reactive Oxigen Species (ROS) production (N-Acetyl-L-cysteine), K+ efflux (Glibenclamide) or phagossome acidification (Bafilomycin). Quantification of IL-1beta and IL-18 in supernatants was performed by ELISA. Our results showed that the production of IL-1beta and IL-18 by THP-1-derived-macrophages stimulated with Pb yeast cells was dependent on NLRP3 and caspase-1 activation, once the presence of their specific inhibitors diminished the production of these cytokines. Furthermore, we found that the major pathways involved in NLRP3 activation, after Pb recognition, were dependent on ROS production and K+ efflux. In conclusion, our results showed that NLRP3 participates in the recognition of Pb yeast cells by macrophages, leading to the activation of the NLRP3-inflammasome and production of IL-1beta and IL-18. Together, these cytokines can induce an inflammatory response against P. brasiliensis, essential for the establishment of the initial inflammatory response and for the development of the subsequent acquired immune response.

Keywords: inflammation, IL-1beta, IL-18, NLRP3, Paracoccidioidomycosis

Procedia PDF Downloads 249