Search results for: live cells imaging
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 5472

Search results for: live cells imaging

3522 Auto Surgical-Emissive Hand

Authors: Abhit Kumar

Abstract:

The world is full of master slave Telemanipulator where the doctor’s masters the console and the surgical arm perform the operations, i.e. these robots are passive robots, what the world needs to focus is that in use of these passive robots we are acquiring doctors for operating these console hence the utilization of the concept of robotics is still not fully utilized ,hence the focus should be on active robots, Auto Surgical-Emissive Hand use the similar concept of active robotics where this anthropomorphic hand focuses on the autonomous surgical, emissive and scanning operation, enabled with the vision of 3 way emission of Laser Beam/-5°C < ICY Steam < 5°C/ TIC embedded in palm of the anthropomorphic hand and structured in a form of 3 way disc. Fingers of AS-EH (Auto Surgical-Emissive Hand) as called, will have tactile, force, pressure sensor rooted to it so that the mechanical mechanism of force, pressure and physical presence on the external subject can be maintained, conversely our main focus is on the concept of “emission” the question arises how all the 3 non related methods will work together that to merged in a single programmed hand, all the 3 methods will be utilized according to the need of the external subject, the laser if considered will be emitted via a pin sized outlet, this radiation is channelized via a thin channel which further connect to the palm of the surgical hand internally leading to the pin sized outlet, here the laser is used to emit radiation enough to cut open the skin for removal of metal scrap or any other foreign material while the patient is in under anesthesia, keeping the complexity of the operation very low, at the same time the TIC fitted with accurate temperature compensator will be providing us the real time feed of the surgery in the form of heat image, this gives us the chance to analyze the level, also ATC will help us to determine the elevated body temperature while the operation is being proceeded, the thermal imaging camera in rooted internally in the AS-EH while also being connected to the real time software externally to provide us live feedback. The ICY steam will provide the cooling effect before and after the operation, however for more utilization of this concept we can understand the working of simple procedure in which If a finger remain in icy water for a long time it freezes the blood flow stops and the portion become numb and isolated hence even if you try to pinch it will not provide any sensation as the nerve impulse did not coordinated with the brain hence sensory receptor did not got active which means no sense of touch was observed utilizing the same concept we can use the icy stem to be emitted via a pin sized hole on the area of concern ,temperature below 273K which will frost the area after which operation can be done, this steam can also be use to desensitized the pain while the operation in under process. The mathematical calculation, algorithm, programming of working and movement of this hand will be installed in the system prior to the procedure, since this AS-EH is a programmable hand it comes with the limitation hence this AS-EH robot will perform surgical process of low complexity only.

Keywords: active robots, algorithm, emission, icy steam, TIC, laser

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3521 Cardiotoxicity Associated with Radiation Therapy: The Role of Bone Marrow Mesenchymal Cells in Improvement of Heart Function

Authors: Isalira Peroba Ramos, Cherley Borba Vieira de Andrade, Grazielle Suhett, Camila Salata, Paulo Cesar Canary, Guilherme Visconde Brasil, Antonio Carlos Campos de Carvalho, Regina Coeli dos Santos Goldenberg

Abstract:

Background: The therapeutic options for patients with cancer now include increasingly complex combinations of medications, radiation therapy (RT), and surgical intervention. Many of these treatments have important potential adverse cardiac effects and are likely to have significant effects on patient outcomes. Cell therapy appears to be promising for the treatment of chronic and degenerative diseases, including cardiomyopathy induced by RT, as the current therapeutic options are insufficient. Aims: To evaluate the potential of bone marrow mesenchymal cells (BMMCs) in radioinduced cardiac damage Methods: Female Wistar rats, 3 months old (Ethics Committee 054/14), were divided into 2 groups, non-treated irradiated group (IR n=15) and irradiated and BMMC treated (IRT n=10). Echocardiography was performed to evaluate heart function. After euthanasia, 3 months post treatment; the left ventricle was removed and prepared for RT-qPCR (VEGF and Pro Collagen I) and histological (picrosirius) analysis. Results: In both groups, 45 days after irradiation, ejection fraction (EF) was in the normal range for these animals (> 70%). However, the BMMC treated group had EF (83.1%±2.6) while the non-treated IR group showed a significant reduction (76.1%±2.6) in relation to the treated group. In addition, we observed an increase in VEGF gene expression and a decrease in Pro Collagen I in IRT when compared to IR group. We also observed by histology that the collagen deposition was reduced in IRT (10.26%±0.83) when compared to IR group (25.29%±0.96). Conclusions: Treatment with BMMCs was able to prevent ejection fraction reduction and collagen deposition in irradiated animals. The increase of VEGF and the decrease of pro collagen I gene expression might explain, at least in part, the cell therapy benefits. All authors disclose no financial or personal relationships with individuals or organizations that could be perceived to bias their work. Sources of funding: FAPERJ, CAPES, CNPq, MCT.

Keywords: mesenchymal cells, radioation, cardiotoxicity, bone marrow

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3520 Efficient Production of Cell-Adhesive Motif From Human Fibronectin Domains to Design a Bio-Functionalized Scaffold for Tissue Engineering

Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

Abstract:

Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

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3519 Relevance Of Cognitive Rehabilitation Amongst Children Having Chronic Illnesses – A Theoretical Analysis

Authors: Pulari C. Milu Maria Anto

Abstract:

Background: Cognitive Rehabilitation/Retraining has been variously used in the research literature to represent non-pharmacological interventions that target the cognitive impairments with the goal of ameliorating cognitive function and functional behaviors to optimize the quality of life. Along with adult’s cognitive impairments, the need to address acquired cognitive impairments (due to any chronic illnesses like CHD - congenital heart diseases or ALL - Acute Lymphoblastic Leukemia) among child populations is inevitable. Also, it has to be emphasized as same we consider the cognitive impairments seen in the children having neurodevelopmental disorders. Methods: All published brain image studies (Hermann, B. et al,2002, Khalil, A. et al., 2004, Follin, C. et al, 2016, etc.) and studies emphasizing cognitive impairments in attention, memory, and/or executive function and behavioral aspects (Henkin, Y. et al,2007, Bellinger, D. C., & Newburger, J. W. (2010), Cheung, Y. T., et al,2016, that could be identified were reviewed. Based on a systematic review of the literature from (2000 -2021) different brain imaging studies, increased risk of neuropsychological and psychosocial impairments are briefly described. Clinical and research gap in the area is discussed. Results:30 papers, both Indian studies and foreign publications (Sage journals, Delhi psychiatry journal, Wiley Online Library, APA PsyNet, Springer, Elsevier, Developmental medicine, and child neurology), were identified. Conclusions: In India, a very limited number of brain imaging studies and neuropsychological studies have done by indicating the cognitive deficits of a child having or undergone chronic illness. None of the studies have emphasized the relevance nor the need of implementingCR among such children, even though its high time to address but still not established yet. The review of the current evidence is to bring out an insight among rehabilitation professionals in establishing a child specific CR and to publish new findings regarding the implementation of CR among such children. Also, this study will be an awareness on considering cognitive aspects of a child having acquired cognitive deficit (due to chronic illness), especially during their critical developmental period.

Keywords: cognitive rehabilitation, neuropsychological impairments, congenital heart diseases, acute lymphoblastic leukemia, epilepsy, and neuroplasticity

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3518 A Comprehensive Approach to Create ‘Livable Streets’ in the Mixed Land Use of Urban Neighborhoods Applying Urban Design Principles Which Will Achieve Quality of Life for Pedestrians

Authors: K. C. Tanuja, Mamatha P. Raj

Abstract:

Urbanisation is happening rapidly all over the world. As population increasing in the urban settlements, its required to provide quality of life to all the inhabitants who live in. Urban design is a place making strategic planning. Urban design principles promote visualising any place environmentally, socially and economically viable. Urban design strategies include building mass, transit development, economic viability and sustenance and social aspects.

Keywords: livable streets, social interaction, pedestrian use, urban design

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3517 Biodegradation of Phenazine-1-Carboxylic Acid by Rhodanobacter sp. PCA2 Proceeds via Decarboxylation and Cleavage of Nitrogen-Containing Ring

Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

Abstract:

Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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3516 IL6/PI3K/mTOR/GFAP Molecular Pathway Role in COVID-19-Induced Neurodegenerative Autophagy, Impacts and Relatives

Authors: Mohammadjavad Sotoudeheian

Abstract:

COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

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3515 In vitro Comparison Study of Biologically Synthesized Cupper-Disulfiram Nanoparticles with Its Free Corresponding Complex as Therapeutic Approach for Breast and Liver Cancer

Authors: Marwa M. Abu-Serie, Marwa M. Eltarahony

Abstract:

The search for reliable, effective, and safe nanoparticles (NPs) as a treatment for cancer is a pressing priority. In this study, Cu-NPs were fabricated by Streptomyces cyaneofuscatus through simultaneous bioreduction strategy of copper nitrate salt. The as-prepared Cu-NPs subjected to structural analysis; energy-dispersive X-ray spectroscopy, elemental mapping, X-ray diffraction, transmission electron microscopy, and ζ-potential. These biological synthesized Cu-NPs were mixed with disulfiram (DS), forming a nanocomplex of Cu-DS with a size of ~135 nm. The prepared nanocomplex (nanoCu-DS) exhibited higher anticancer activity than that of free complex of DS-Cu, Cu-NPs, and DS alone. This was illustrated by the lowest IC50 of nanoCu-DS (< 4 µM) against human breast and liver cancer cell lines comparing with DS-Cu, Cu-NPs, and DS (~8, 22.98-33.51 and 11.95-14.86, respectively). Moreover, flow cytometric analysis confirmed that higher apoptosis percentage range of nanoCu-DS-treated in MDA-MB 231, MCF-7, Huh-7, and HepG-2 cells (51.24-65.28%) than free complex of Cu-DS ( < 4.5%). Regarding inhibition potency of liver and breast cancer cell migration, no significant difference was recorded between free and nanocomplex. Furthermore, nanoCu-DS suppressed gene expression of β-catenine, Akt, and NF-κB and upregulated p53 expression (> 3, >15, > 5 and ≥ 3 folds, respectively) more efficiently than free complex (all ~ 1 fold) in MDA-MB 231 and Huh-7 cells. Our finding proved this prepared nano complex has a powerful anticancer activity relative to free complex, thereby offering a promising cancer treatment.

Keywords: biologically prepared Cu-NPs, breast cancer cell lines, liver cancer cell lines, nanoCu- disulfiram

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3514 Development of Three-Dimensional Bio-Reactor Using Magnetic Field Stimulation to Enhance PC12 Cell Axonal Extension

Authors: Eiji Nakamachi, Ryota Sakiyama, Koji Yamamoto, Yusuke Morita, Hidetoshi Sakamoto

Abstract:

The regeneration of injured central nerve network caused by the cerebrovascular accidents is difficult, because of poor regeneration capability of central nerve system composed of the brain and the spinal cord. Recently, new regeneration methods such as transplant of nerve cells and supply of nerve nutritional factor were proposed and examined. However, there still remain many problems with the canceration of engrafted cells and so on and it is strongly required to establish an efficacious treating method of a central nerve system. Blackman proposed the electromagnetic stimulation method to enhance the axonal nerve extension. In this study, we try to design and fabricate a new three-dimensional (3D) bio-reactor, which can load a uniform AC magnetic field stimulation on PC12 cells in the extracellular environment for enhancement of an axonal nerve extension and 3D nerve network generation. Simultaneously, we measure the morphology of PC12 cell bodies, axons, and dendrites by the multiphoton excitation fluorescence microscope (MPM) and evaluate the effectiveness of the uniform AC magnetic stimulation to enhance the axonal nerve extension. Firstly, we designed and fabricated the uniform AC magnetic field stimulation bio-reactor. For the AC magnetic stimulation system, we used the laminated silicon steel sheets for a yoke structure of 3D chamber, which had a high magnetic permeability. Next, we adopted the pole piece structure and installed similar specification coils on both sides of the yoke. We searched an optimum pole piece structure using the magnetic field finite element (FE) analyses and the response surface methodology. We confirmed that the optimum 3D chamber structure showed a uniform magnetic flux density in the PC12 cell culture area by using FE analysis. Then, we fabricated the uniform AC magnetic field stimulation bio-reactor by adopting analytically determined specifications, such as the size of chamber and electromagnetic conditions. We confirmed that measurement results of magnetic field in the chamber showed a good agreement with FE results. Secondly, we fabricated a dish, which set inside the uniform AC magnetic field stimulation of bio-reactor. PC12 cells were disseminated with collagen gel and could be 3D cultured in the dish. The collagen gel were poured in the dish. The collagen gel, which had a disk shape of 6 mm diameter and 3mm height, was set on the membrane filter, which was located at 4 mm height from the bottom of dish. The disk was full filled with the culture medium inside the dish. Finally, we evaluated the effectiveness of the uniform AC magnetic field stimulation to enhance the nurve axonal extension. We confirmed that a 6.8 increase in the average axonal extension length of PC12 under the uniform AC magnetic field stimulation at 7 days culture in our bio-reactor, and a 24.7 increase in the maximum axonal extension length. Further, we confirmed that a 60 increase in the number of dendrites of PC12 under the uniform AC magnetic field stimulation. Finally, we confirm the availability of our uniform AC magnetic stimulation bio-reactor for the nerve axonal extension and the nerve network generation.

Keywords: nerve regeneration, axonal extension , PC12 cell, magnetic field, three-dimensional bio-reactor

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3513 The Effect of High-Pressure Processing on the Inactivation of Saccharomyces cerevisiae in Different Concentration of Manuka Honey and Its Relation with ° Brix

Authors: Noor Akhmazillah Fauzi, Mohammed Mehdi Farid, Filipa V. Silva

Abstract:

The aim of this paper is to investigate if different concentration of Manuka honey (as a model food) has a major influence on the inactivation of Saccharomyces cerevisiae (as the testing microorganism) after subjecting it to HPP. Honey samples with different sugar concentrations (20, 30, 40, 50, 60 and 70 °Brix) were prepared aseptically using sterilized distilled water. No dilution of honey was made for the 80 °Brix sample. For the 0 °Brix sample (control), sterilized distilled water was used. Thermal treatment at 55 °C for 10 min (conventionally applied in honey pasteurisation in industry) was carried out for comparison purpose. S. cerevisiae cell numbers in honey samples were established before and after each HPP and thermal treatment. The number of surviving cells was determined after a proper dilution of the untreated and treated samples by the viable plate count method. S. cerevisiae cells, in different honey concentrations (0 to 80 °Brix), subjected to 600 MPa (at ambient temperature) showed an increasing resistance to inactivation with °Brix. A significant correlation (p < 0.05) between cell reduction and °Brix was found. Cell reduction in high pressure-treated samples varied linearly with °Brix (R2 > 0.9), confirming that the baroprotective effect of the food is due to sugar content. This study has practical implications in establishing efficient process design for commercial manufacturing of high sugar food products and on the potential use of HPP for such products.

Keywords: high pressure processing, honey, Saccharomyces cerevisiae, °Brix

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3512 Efficient Delivery of Biomaterials into Living Organism by Using Noble Metal Nanowire Injector

Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

Abstract:

Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

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3511 Transmission Line Inspection Using Drones

Authors: Jae Kyung Lee, Joon Young Park

Abstract:

Maintenance on power transmission lines requires a lot of works. Sometimes they should be maintained on live-line environment with high altitude. Therefore, there always exist risks of falling from height and electric shock. To decline those risks, drones are recently applying on the electric power industry. This paper presents new operational technology while inspecting power transmission line. This paper also describes a technique for creating a flight path of a drone for transmission line inspection and a technique for controlling the drones of different types. Its technical and economical feasibilities have confirmed through experiments.

Keywords: drones, transmission line, inspection, control system

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3510 Validation of an Impedance-Based Flow Cytometry Technique for High-Throughput Nanotoxicity Screening

Authors: Melanie Ostermann, Eivind Birkeland, Ying Xue, Alexander Sauter, Mihaela R. Cimpan

Abstract:

Background: New reliable and robust techniques to assess biological effects of nanomaterials (NMs) in vitro are needed to speed up safety analysis and to identify key physicochemical parameters of NMs, which are responsible for their acute cytotoxicity. The central aim of this study was to validate and evaluate the applicability and reliability of an impedance-based flow cytometry (IFC) technique for the high-throughput screening of NMs. Methods: Eight inorganic NMs from the European Commission Joint Research Centre Repository were used: NM-302 and NM-300k (Ag: 200 nm rods and 16.7 nm spheres, respectively), NM-200 and NM- 203 (SiO₂: 18.3 nm and 24.7 nm amorphous, respectively), NM-100 and NM-101 (TiO₂: 100 nm and 6 nm anatase, respectively), and NM-110 and NM-111 (ZnO: 147 nm and 141 nm, respectively). The aim was to assess the biological effects of these materials on human monoblastoid (U937) cells. Dispersions of NMs were prepared as described in the NANOGENOTOX dispersion protocol and cells were exposed to NMs at relevant concentrations (2, 10, 20, 50, and 100 µg/mL) for 24 hrs. The change in electrical impedance was measured at 0.5, 2, 6, and 12 MHz using the IFC AmphaZ30 (Amphasys AG, Switzerland). A traditional toxicity assay, Trypan Blue Dye Exclusion assay, and dark-field microscopy were used to validate the IFC method. Results: Spherical Ag particles (NM-300K) showed the highest toxic effect on U937 cells followed by ZnO (NM-111 ≥ NM-110) particles. Silica particles were moderate to non-toxic at all used concentrations under these conditions. A higher toxic effect was seen with smaller sized TiO2 particles (NM-101) compared to their larger analogues (NM-100). No interferences between the IFC and the used NMs were seen. Uptake and internalization of NMs were observed after 24 hours exposure, confirming actual NM-cell interactions. Conclusion: Results collected with the IFC demonstrate the applicability of this method for rapid nanotoxicity assessment, which proved to be less prone to nano-related interference issues compared to some traditional toxicity assays. Furthermore, this label-free and novel technique shows good potential for up-scaling in directions of an automated high-throughput screening and for future NM toxicity assessment. This work was supported by the EC FP7 NANoREG (Grant Agreement NMP4-LA-2013-310584), the Research Council of Norway, project NorNANoREG (239199/O70), the EuroNanoMed II 'GEMN' project (246672), and the UH-Nett Vest project.

Keywords: cytotoxicity, high-throughput, impedance, nanomaterials

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3509 New Active Dioxin Response Element Sites in Regulatory Region of Human and Viral Genes

Authors: Ilya B. Tsyrlov, Dmitry Y. Oshchepkov

Abstract:

A computational search for dioxin response elements (DREs) in genes of proteins comprising the Ah receptor (AhR) cytosolic core complex was performed by highly efficient tool SITECON. Eventually, the following number of new DREs in 5’flanking region was detected by SITECON: one in AHR gene, five in XAP2, eight in HSP90AA1, and three in HSP90AB1 genes. Numerous DREs found in genes of AhR and AhR cytosolic complex members would shed a light on potential mechanisms of expression, the stoichiometry of unliganded AhR core complex, and its degradation vs biosynthesis dynamics resulted from treatment of target cells with the AhR most potent ligand, 2,3,7,8-TCDD. With human viruses, reduced susceptibility to TCDD of geneencoding HIV-1 P247 was justified by the only potential DRE determined in gag gene encoding HIV-1 P24 protein, whereas the regulatory region of CMV genes encoding IE gp/UL37 has five potent DRE, 1.65 kb/UL36 – six DRE, pp65 and pp71 – each has seven DRE, and pp150 – ten DRE. Also, from six to eight DRE were determined with SITECON in the regulatory region of HSV-1 IE genes encoding tegument proteins, UL36 and UL37, and of UL19 gene encoding bindingglycoprotein C (gC). So, TCDD in the low picomolar range may activate in human cells AhR: Arnt transcription pathway that triggers CMV and HSV-1 reactivation by binding to numerous promoter DRE within immediate-early (IE) genes UL37 and UL36, thus committing virus to the lytic cycle.

Keywords: dioxin response elements, Ah receptor, AhR: Arnt transcription pathway, human and viral genes

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3508 Anti-proliferative Activity and HER2 Receptor Expression Analysis of MCF-7 (Breast Cancer Cell) Cells by Plant Extract Coleus Barbatus (Andrew)

Authors: Anupalli Roja Rani, Pavithra Dasari

Abstract:

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

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

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3507 Diagnostic Accuracy in the Detection of Cervical Lymph Node Metastases in Head and Neck Squamous Cell Carcinoma Patients: A Comparison of Sonography, CT, PET/CT and MRI

Authors: Di Luo, Maria Buchberger, Anja Pickhard

Abstract:

Objectives: The purpose of this study was to assess and compare the diagnostic accuracy of four common morphological approaches, including sonography, computed tomography (CT), positron emission tomography/computed tomography (PET/CT), and magnetic resonance imaging (MRI) for the evaluation of cervical lymph node metastases in head and neck squamous cell carcinoma (HNSCC) patients. Material and Methods: Included in this retrospective study were 26 patients diagnosed with HNSCC between 2010 and 2011 who all underwent sonography, CT, PET/CT, and MRI imaging before neck dissection. Morphological data were compared to the corresponding histopathological results. Statistical analysis was performed with SPSS statistic software (version 26.0), calculating sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy for detection of cervical lymph node metastases. Results: The 5-year survival rate of the patient collective was 55.5%.Risk factors for survival included initial primary tumor stage, initial lymph node stage, initial metastasis status, and therapeutic approaches. Cox regression showed initial metastasis status(HR 8.671, 95%CI 1.316-57.123, p=0.025) and therapeutic approaches(HR 6.699, 95%CI 1.746-25.700, p=0.006)to be independent predictive risk factors for survival. Sensitivity was highest for MRI (96% compared to 85% for sonography and 89% for CT and PET/CT). Specificity was comparable with 95 % for CT and 98 % for sonography and PET/CT, but only 68% for MRI. While the MRI showed the least PPV (34%) compared to all other methods (85% for sonography,75% for CT, and 86% for PET/CT), the NPV was comparable in all methods(98-99%). The overall accuracy of cervical lymph node metastases detection was comparable for sonography, CT, and PET/CT with 96%,97%,94%, respectively, while MRI had only 72% accuracy. Conclusion: Since the initial status of metastasis is an independent predictive risk factor for patients’ survival, efficient detection is crucial to plan adequate therapeutic approaches. Sonography, CT, and PET/CT have better diagnostic accuracy than MRI for the evaluation of cervical lymph node metastases in HNSCC patients.

Keywords: cervical lymph node metastases, diagnostic accuracy, head and neck squamous carcinoma, risk factors, survival

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3506 N-Glycosylation in the Green Microalgae Chlamydomonas reinhardtii

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

Abstract:

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

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

Procedia PDF Downloads 175
3505 Regulation of Desaturation of Fatty Acid and Triglyceride Synthesis by Myostatin through Swine-Specific MEF2C/miR222/SCD5 Pathway

Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

Abstract:

Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

Procedia PDF Downloads 129
3504 Desirable Fatty Acids in Meat of Cattle Fed Different Levels of Lipid-Based Diets

Authors: Tiago N. P. Valente, Erico S. Lima, Roberto O. Roça

Abstract:

Introduction: Research has stimulated animal production studies on solutions to decrease the level of saturated fatty acids and increase unsaturated in foods of animal origin. The objective of this study was to determine the effect of the dietary inclusion of lipid-based diets on the fatty acid profiles from finishing cattle. Materials and Methods: The study was carried out in the Chapéu de Couro Farm in Aguaí/SP, Brazil. A group of 39 uncastrated Nellore cattle. Mean age of the animals was 36 months, and initial mean live weight was 494.1 ± 10.1. Animals were randomly assigned to one of three treatments, based on dry matter: feed with control diet 2.50% cottonseed, feed with 11.50% cottonseed, and feed with 3.13% cottonseed added of 1.77% protected lipid. Forage:concentrate ratio was 50:50 on a dry matter basis. Sugar cane chopped was used as forage. After 63 days mean final live weight was 577.01 kg ± 11.34. After slaughter, carcasses were identified and divided into two halves that were kept in a cold chamber for 24 hours at 2°C. Then, part of the M. longissimus thoracis of each animal was removed between the 12th and 13th rib of the left half carcass. The samples steaks were 2.5 cm thick and were identified and stored frozen in a freezer at -18°C. The analysis of methyl esters of fatty acids was carried out in a gas chromatograph. Desirable fatty acids (FADes) were determined by the sum of unsaturated fatty acids and stearic acid (C18:0). Results and Discussion: No differences (P>0.05) between the diets for the proportion of FADes in the meat of the animals in this study, according to the lipid sources used. The inclusion of protected fat or cottonseed in the diet did not change the proportion of FADes in the meat. The proportion mean of FADes in meat in the present study were: as pentadecanoic acid (C15:1 = 0.29%), palmitoleic acid (C16:1 = 4.26%), heptadecanoic acid (C17:1 = 0.07%), oleic acid (C18:1n9c = 37.32%), γ-linolenic acid (0.94%) and α-linolenic acid (1.04%), elaidic acid (C18:1n9t = 0.50%), eicosatrienoic acid (C20:3n3 = 0.03%), eicosapentaenoic acid (C20:5n3 = 0.04%), erucic acid (C22:1n9 = 0.89%), docosadienoic acid (C22:2 = 0.04%) and stearic acid (C18:0 = 21.53%). Conclusions: The add the cottonseed or protected lipid in diet is not affected fatty acids profiles the desirable fatty acids in meat. Acknowledgements: IFGoiano, FAPEG and CNPq (Brazil).

Keywords: beef quality, cottonseed, protected fat, unsaturated fatty acids

Procedia PDF Downloads 290
3503 Application of Infrared Thermal Imaging, Eye Tracking and Behavioral Analysis for Deception Detection

Authors: Petra Hypšová, Martin Seitl

Abstract:

One of the challenges of forensic psychology is to detect deception during a face-to-face interview. In addition to the classical approaches of monitoring the utterance and its components, detection is also sought by observing behavioral and physiological changes that occur as a result of the increased emotional and cognitive load caused by the production of distorted information. Typical are changes in facial temperature, eye movements and their fixation, pupil dilation, emotional micro-expression, heart rate and its variability. Expanding technological capabilities have opened the space to detect these psychophysiological changes and behavioral manifestations through non-contact technologies that do not interfere with face-to-face interaction. Non-contact deception detection methodology is still in development, and there is a lack of studies that combine multiple non-contact technologies to investigate their accuracy, as well as studies that show how different types of lies produced by different interviewers affect physiological and behavioral changes. The main objective of this study is to apply a specific non-contact technology for deception detection. The next objective is to investigate scenarios in which non-contact deception detection is possible. A series of psychophysiological experiments using infrared thermal imaging, eye tracking and behavioral analysis with FaceReader 9.0 software was used to achieve our goals. In the laboratory experiment, 16 adults (12 women, 4 men) between 18 and 35 years of age (SD = 4.42) were instructed to produce alternating prepared and spontaneous truths and lies. The baseline of each proband was also measured, and its results were compared to the experimental conditions. Because the personality of the examiner (particularly gender and facial appearance) to whom the subject is lying can influence physiological and behavioral changes, the experiment included four different interviewers. The interviewer was represented by a photograph of a face that met the required parameters in terms of gender and facial appearance (i.e., interviewer likability/antipathy) to follow standardized procedures. The subject provided all information to the simulated interviewer. During follow-up analyzes, facial temperature (main ROIs: forehead, cheeks, the tip of the nose, chin, and corners of the eyes), heart rate, emotional expression, intensity and fixation of eye movements and pupil dilation were observed. The results showed that the variables studied varied with respect to the production of prepared truths and lies versus the production of spontaneous truths and lies, as well as the variability of the simulated interviewer. The results also supported the assumption of variability in physiological and behavioural values during the subject's resting state, the so-called baseline, and the production of prepared and spontaneous truths and lies. A series of psychophysiological experiments provided evidence of variability in the areas of interest in the production of truths and lies to different interviewers. The combination of technologies used also led to a comprehensive assessment of the physiological and behavioral changes associated with false and true statements. The study presented here opens the space for further research in the field of lie detection with non-contact technologies.

Keywords: emotional expression decoding, eye-tracking, functional infrared thermal imaging, non-contact deception detection, psychophysiological experiment

Procedia PDF Downloads 98
3502 The Cell Viability Study of Extracts of Bark, Flowers, Leaves and Seeds of Indian Dhak Tree, Flame of Forest

Authors: Madhavi S. Apte, Milind Bhitre

Abstract:

In pharmaceutical research and new drug development, medicinal plants have important roles. Similarly, Indian dhak tree belonging to family Fabaceae has been widely used in the traditional Indian medical system of ‘Ayurveda’ for the treatment of a variety of ailments. Hence the cell viability study was undertaken to evaluate and compare the activity of extracts of various parts like flower, bark, leaf, seed by conducting MTT assay method along with other pharmacognostical studies. The methanolic extracts of bark, flowers, leaves, and seeds were used for the study. The cell viability MTT assay was performed using the standard operating procedures. The extracts were dissolved in DMSO and serially diluted with complete medium to get the concentrations range of test concentration. DMSO concentration was kept < 0.1% in all the samples. HUVEC cells maintained in appropriate conditions were seeded in 96 well plates and treated with different concentrations of the test samples and incubated at 37°C, 5% CO₂ for 96 hours. MTT reagent was added to the wells and incubated for 4 hours; the dark blue formazan product formed by the cells was dissolved in DMSO under a safety cabinet and read at 550nm. Percentage inhibitions were calculated and plotted with the concentrations used to calculate the IC50 values. The bark, flower, leaves and seed extracts have shown the cytotoxicity activity and can be further studied for antiangiogenesis activity.

Keywords: pharmacognosy, Cell viability, MTT assay, anti-angiogenesis

Procedia PDF Downloads 295
3501 The Resistance of Fish Outside of Water Medium

Authors: Febri Ramadhan

Abstract:

Water medium is a vital necessity for the survival of fish. Fish can survive inside/outside of water medium within a certain time. By knowing the level of survival fish at outside of water medium, a person can transport the fish to a place with more efficiently. Transport of live fish from one place to another can be done with wet and dry media system. In this experiment the treatment-given the observed differences in fish species. This experiment aimed to test the degree of resilience of fish out of water media. Based on the ANOVA table is obtained, it can be concluded that the type of fish affects the level of resilience of fish outside the water (Fhit> Ftab).

Keywords: fish, transport, retention rate, fish resiliance

Procedia PDF Downloads 334
3500 Inhibitory Effects of Crocin from Crocus sativus L. on Cell Proliferation of a Medulloblastoma Human Cell Line

Authors: Kyriaki Hatziagapiou, Eleni Kakouri, Konstantinos Bethanis, Alexandra Nikola, Eleni Koniari, Charalabos Kanakis, Elias Christoforides, George Lambrou, Petros Tarantilis

Abstract:

Medulloblastoma is a highly invasive tumour, as it tends to disseminate throughout the central nervous system early in its course. Despite the high 5-year-survival rate, a significant number of patients demonstrate serious long- or short-term sequelae (e.g., myelosuppression, endocrine dysfunction, cardiotoxicity, neurological deficits and cognitive impairment) and higher mortality rates, unrelated to the initial malignancy itself but rather to the aggressive treatment. A strong rationale exists for the use of Crocus sativus L (saffron) and its bioactive constituents (crocin, crocetin, safranal) as pharmaceutical agents, as they exert significant health-promoting properties. Crocins are water soluble carotenoids. Unlike other carotenoids, crocins are highly water-soluble compounds, with relatively low toxicity as they are not stored in adipose and liver tissues. Crocins have attracted wide attention as promising anti-cancer agents, due to their antioxidant, anti-inflammatory, and immunomodulatory effects, interference with transduction pathways implicated in tumorigenesis, angiogenesis, and metastasis (disruption of mitotic spindle assembly, inhibition of DNA topoisomerases, cell-cycle arrest, apoptosis or cell differentiation) and sensitization of cancer cells to radiotherapy and chemotherapy. The current research aimed to study the potential cytotoxic effect of crocins on TE671 medulloblastoma cell line, which may be useful in the optimization of existing and development of new therapeutic strategies. Crocins were extracted from stigmas of saffron in ultrasonic bath, using petroleum-ether, diethylether and methanol 70%v/v as solvents and the final extract was lyophilized. Identification of crocins according to high-performance liquid chromatography (HPLC) analysis was determined comparing the UV-vis spectra and the retention time (tR) of the peaks with literature data. For the biological assays crocin was diluted to nuclease and protease free water. TE671 cells were incubated with a range of concentrations of crocins (16, 8, 4, 2, 1, 0.5 and 0.25 mg/ml) for 24, 48, 72 and 96 hours. Analysis of cell viability after incubation with crocins was performed with Alamar Blue viability assay. The active ingredient of Alamar Blue, resazurin, is a blue, nontoxic, cell permeable compound virtually nonfluorescent. Upon entering cells, resazurin is reduced to a pink and fluorescent molecule, resorufin. Viable cells continuously convert resazurin to resorufin, generating a quantitative measure of viability. The colour of resorufin was quantified by measuring the absorbance of the solution at 600 nm with a spectrophotometer. HPLC analysis indicated that the most abundant crocins in our extract were trans-crocin-4 and trans-crocin-3. Crocins exerted significant cytotoxicity in a dose and time-dependent manner (p < 0.005 for exposed cells to any concentration at 48, 72 and 96 hours versus cells not exposed); as their concentration and time of exposure increased, the reduction of resazurin to resofurin decreased, indicating reduction in cell viability. IC50 values for each time point were calculated ~3.738, 1.725, 0.878 and 0.7566 mg/ml at 24, 48, 72 and 96 hours, respectively. The results of our study could afford the basis of research regarding the use of natural carotenoids as anticancer agents and the shift to targeted therapy with higher efficacy and limited toxicity. Acknowledgements: The research was funded by Fellowships of Excellence for Postgraduate Studies IKY-Siemens Programme.

Keywords: crocetin, crocin, medulloblastoma, saffron

Procedia PDF Downloads 215
3499 Rapid, Label-Free, Direct Detection and Quantification of Escherichia coli Bacteria Using Nonlinear Acoustic Aptasensor

Authors: Shilpa Khobragade, Carlos Da Silva Granja, Niklas Sandström, Igor Efimov, Victor P. Ostanin, Wouter van der Wijngaart, David Klenerman, Sourav K. Ghosh

Abstract:

Rapid, label-free and direct detection of pathogenic bacteria is critical for the prevention of disease outbreaks. This paper for the first time attempts to probe the nonlinear acoustic response of quartz crystal resonator (QCR) functionalized with specific DNA aptamers for direct detection and quantification of viable E. coli KCTC 2571 bacteria. DNA aptamers were immobilized through biotin and streptavidin conjugation, onto the gold surface of QCR to capture the target bacteria and the detection was accomplished by shift in amplitude of the peak 3f signal (3 times the drive frequency) upon binding, when driven near fundamental resonance frequency. The developed nonlinear acoustic aptasensor system demonstrated better reliability than conventional resonance frequency shift and energy dissipation monitoring that were recorded simultaneously. This sensing system could directly detect 10⁽⁵⁾ cells/mL target bacteria within 30 min or less and had high specificity towards E. coli KCTC 2571 bacteria as compared to the same concentration of S.typhi bacteria. Aptasensor response was observed for the bacterial suspensions ranging from 10⁽⁵⁾-10⁽⁸⁾ cells/mL. Conclusively, this nonlinear acoustic aptasensor is simple to use, gives real-time output, cost-effective and has the potential for rapid, specific, label-free direction detection of bacteria.

Keywords: acoustic, aptasensor, detection, nonlinear

Procedia PDF Downloads 564
3498 Enhancing Early Detection of Coronary Heart Disease Through Cloud-Based AI and Novel Simulation Techniques

Authors: Md. Abu Sufian, Robiqul Islam, Imam Hossain Shajid, Mahesh Hanumanthu, Jarasree Varadarajan, Md. Sipon Miah, Mingbo Niu

Abstract:

Coronary Heart Disease (CHD) remains a principal cause of global morbidity and mortality, characterized by atherosclerosis—the build-up of fatty deposits inside the arteries. The study introduces an innovative methodology that leverages cloud-based platforms like AWS Live Streaming and Artificial Intelligence (AI) to early detect and prevent CHD symptoms in web applications. By employing novel simulation processes and AI algorithms, this research aims to significantly mitigate the health and societal impacts of CHD. Methodology: This study introduces a novel simulation process alongside a multi-phased model development strategy. Initially, health-related data, including heart rate variability, blood pressure, lipid profiles, and ECG readings, were collected through user interactions with web-based applications as well as API Integration. The novel simulation process involved creating synthetic datasets that mimic early-stage CHD symptoms, allowing for the refinement and training of AI algorithms under controlled conditions without compromising patient privacy. AWS Live Streaming was utilized to capture real-time health data, which was then processed and analysed using advanced AI techniques. The novel aspect of our methodology lies in the simulation of CHD symptom progression, which provides a dynamic training environment for our AI models enhancing their predictive accuracy and robustness. Model Development: it developed a machine learning model trained on both real and simulated datasets. Incorporating a variety of algorithms including neural networks and ensemble learning model to identify early signs of CHD. The model's continuous learning mechanism allows it to evolve adapting to new data inputs and improving its predictive performance over time. Results and Findings: The deployment of our model yielded promising results. In the validation phase, it achieved an accuracy of 92% in predicting early CHD symptoms surpassing existing models. The precision and recall metrics stood at 89% and 91% respectively, indicating a high level of reliability in identifying at-risk individuals. These results underscore the effectiveness of combining live data streaming with AI in the early detection of CHD. Societal Implications: The implementation of cloud-based AI for CHD symptom detection represents a significant step forward in preventive healthcare. By facilitating early intervention, this approach has the potential to reduce the incidence of CHD-related complications, decrease healthcare costs, and improve patient outcomes. Moreover, the accessibility and scalability of cloud-based solutions democratize advanced health monitoring, making it available to a broader population. This study illustrates the transformative potential of integrating technology and healthcare, setting a new standard for the early detection and management of chronic diseases.

Keywords: coronary heart disease, cloud-based ai, machine learning, novel simulation techniques, early detection, preventive healthcare

Procedia PDF Downloads 63
3497 Nanoporous Activated Carbons for Fuel Cells and Supercapacitors

Authors: A. Volperts, G. Dobele, A. Zhurinsh, I. Kruusenberg, A. Plavniece, J. Locs

Abstract:

Nowadays energy consumption constantly increases and development of effective and cheap electrochemical sources of power, such as fuel cells and electrochemical capacitors, is topical. Due to their high specific power, charge and discharge rates, working lifetime supercapacitor based energy accumulation systems are more and more extensively being used in mobile and stationary devices. Lignocellulosic materials are widely used as precursors and account for around 45% of the total raw materials used for the manufacture of activated carbon which is the most suitable material for supercapacitors. First part of our research is devoted to study of influence of main stages of wood thermochemical activation parameters on activated carbons porous structure formation. It was found that the main factors governing the properties of carbon materials are specific surface area, volume and pore size distribution, particles dispersity, ash content and oxygen containing groups content. Influence of activated carbons attributes on capacitance and working properties of supercapacitor are demonstrated. The correlation between activated carbons porous structure indices and electrochemical specifications of supercapacitors with electrodes made from these materials has been determined. It is shown that if synthesized activated carbons are used in supercapacitors then high specific capacitances can be reached – more than 380 F/g in 4.9M sulfuric acid based electrolytes and more than 170 F/g in 1 M tetraethylammonium tetrafluoroborate in acetonitrile electrolyte. Power specifications and minimal price of H₂-O₂ fuel cells are limited by the expensive platinum-based catalysts. The main direction in development of non-platinum catalysts for the oxygen reduction is the study of cheap porous carbonaceous materials which can be obtained by the pyrolysis of polymers including renewable biomass. It is known that nitrogen atoms in carbon materials to a high degree determine properties of the doped activated carbons, such as high electrochemical stability, hardness, electric resistance, etc. The lack of sufficient knowledge on the doping of the carbon materials calls for the ongoing researches of properties and structure of modified carbon matrix. In the second part of this study, highly porous activated carbons were synthesized using alkali thermochemical activation from wood, cellulose and cellulose production residues – craft lignin and sewage sludge. Activated carbon samples were doped with dicyandiamide and melamine for the application as fuel cell cathodes. Conditions of nitrogen introduction (solvent, treatment temperature) and its content in the carbonaceous material, as well as porous structure characteristics, such as specific surface and pore size distribution, were studied. It was found that efficiency of doping reaction depends on the elemental oxygen content in the activated carbon. Relationships between nitrogen content, porous structure characteristics and electrodes electrochemical properties are demonstrated.

Keywords: activated carbons, low-temperature fuel cells, nitrogen doping, porous structure, supercapacitors

Procedia PDF Downloads 117
3496 Rumen Epithelium Development of Bovine Fetuses and Newborn Calves

Authors: Juliana Shimara Pires Ferrão, Letícia Palmeira Pinto, Francisco Palma Rennó, Francisco Javier Hernandez Blazquez

Abstract:

The ruminant stomach is a complex and multi-chambered organ. Although the true stomach (abomasum) is fully differentiated and functional at birth, the same does not occur with the rumen chamber. At this moment, rumen papillae are small or nonexistent. The papillae only fully develop after weaning and during calf growth. Papillae development and ruminal epithelium specialization during the fetus growth and at birth must be two interdependent processes that will prepare the rumen to adapt to ruminant adult feeding. The microscopic study of rumen epithelium at these early phases of life is important to understand how this structure prepares the rumen to deal with the following weaning processes and its functional activation. Samples of ruminal mucosa of bovine fetuses (110- and 150 day-old) and newborn calves were collected (dorsal and ventral portions) and processed for light and electron microscopy and immunohistochemistry. The basal cell layer of the stratified pavimentous epithelium present in different ruminal portions of the fetuses was thicker than the same portions of newborn calves. The superficial and intermediate epithelial layers of 150 day-old fetuses were thicker than those found in the other 2 studied ages. At this age (150 days), dermal papillae begin to invade the intermediate epithelial layer which gradually disappears in newborn calves. At birth, the ruminal papillae project from the epithelial surface, probably by regression of the epithelial cells (transitory cells) surrounding the dermal papillae. The PCNA cell proliferation index (%) was calculated for all epithelial samples. Fetuses 150 day-old showed increased cell proliferation in basal cell layer (Dorsal Portion: 84.2%; Ventral Portion: 89.8%) compared to other ages studied. Newborn calves showed an intermediate index (Dorsal Portion: 65.1%; Ventral Portion: 48.9%), whereas 110 day-old fetuses had the lowest proliferation index (Dorsal Portion: 57.2%; Ventral Portion: 20.6%). Regarding the transitory epithelium, 110 day-old fetuses showed the lowest proliferation index (Dorsal Portion: 44.6%; Ventral Portion: 20.1%), 150 day-old fetuses showed an intermediate proliferation index (Dorsal Portion: 57.5%; Ventral Portion: 71.1%) and newborn calves presented a higher proliferation index (Dorsal Portion: 75.1%; Ventral Portion: 19.6%). Under TEM, the 110- and 150 day-old fetuses presented thicker and poorly organized basal cell layer, with large nuclei and dense cytoplasm. In newborn calves, the basal cell layer was more organized and with fewer layers, but typically similar in both regions of the rumen. For the transitory epithelium, fetuses displayed larger cells than those found in newborn calves with less electrondense cytoplasm than that found in the basal cells. The ruminal dorsal portion has an overall higher cell proliferation rate than the ventral portion. Thus we can infer that the dorsal portion may have a higher cell activity than the ventral portion during ruminal development. Moreover, the basal cell layer is thicker in the 110- and 150 day-old fetuses than in the newborn calves. The transitory epithelium, which is much reduced, at birth may have a structural support function of the developing dermal papillae. When it regresses or is sheared off, the papillae are “carved out” from the surrounding epithelial layer.

Keywords: bovine, calf, epithelium, fetus, hematoxylin-eosin, immunohistochemistry, TEM, Rumen

Procedia PDF Downloads 385
3495 Design of RF Generator and Its Testing in Heating of Nickel Ferrite Nanoparticles

Authors: D. Suman, M. Venkateshwara Rao

Abstract:

Cancer is a disease caused by an uncontrolled division of abnormal cells in a part of the body, which is affecting millions of people leading to death. Even though there have been tremendous developments taken place over the last few decades the effective therapy for cancer is still not a reality. The existing techniques of cancer therapy are chemotherapy and radio therapy which are having their limitations in terms of the side effects, patient discomfort, radiation hazards and the localization of treatment. This paper describes a novel method for cancer therapy by using RF-hyperthermia application of nanoparticles. We have synthesized ferromagnetic nanoparticles and characterized by using XRD and TEM. These nanoparticles after the biocompatibility studies will be injected in to the body with a suitable tracer element having affinity to the specific tumor site. When RF energy is applied to the nanoparticles at the tumor site it produces heat of excess room temperature and nearly 41-45°C is sufficient to kill the tumor cells. We have designed a RF source generator provided with a temperature feedback controller to control the radiation induced temperature of the tumor site. The temperature control is achieved through a negative feedback mechanism of the thermocouple and a relay connected to the power source of the RF generator. This method has advantages in terms of its effect like localized therapy, less radiation, and no side effects. It has several challenges in designing the RF source provided with coils suitable for the tumour site, biocompatibility of the nanomaterials, cooling system design for the RF coil. If we can overcome these challenges this method will be a huge benefit for the society.

Keywords: hyperthermia, cancer therapy, RF source generator, nanoparticles

Procedia PDF Downloads 457
3494 Detection Kit of Type 1 Diabetes Mellitus with Autoimmune Marker GAD65 (Glutamic Acid Decarboxylase)

Authors: Aulanni’am Aulanni’am

Abstract:

Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

Procedia PDF Downloads 266
3493 Comparison Serum Vitamin D by Geographic between the Highland and Lowland Schizophrenic Patient in the Sumatera Utara

Authors: Novita Linda Akbar, Elmeida Effendy, Mustafa M. Amin

Abstract:

Background: The most common of psychotic disorders is schizophrenia. Vitamin D is made from sunlight, and in the skin from UVB radiation from sunlight. If people with Vitamin D deficiency is common severe mental illness such as schizophrenia.Schizophrenia is a chronic mental illness characterised by positive symptoms and negatives symptoms, such as hallucinations and delusions, flat affect and lack of motivation we can found. In patients with Schizophrenia maybe have several environmental risk factors for schizophrenia, such as season of birth, latitude, and climate has been linked to vitamin D deficiency. There is also relationship between the risk of schizophrenia and latitude, and with an increased incidence rate of schizophrenia seen at a higher latitude. Methods: This study was an analytical study, conducted in BLUD RS Jiwa Propinsi Sumatera Utara and RSUD Deli Serdang, the period in May 2016 and ended in June 2016 with a sample of the study 60 sample (20 patients live in the Highland and Lowland, 20 healthy controls). Inclusion criteria were schizophrenic patients both men and women, aged between 18 to 60 years old, acute phase no agitation or abstinence antipsychotic drugs for two weeks, live in the Highland and Lowland, and willing to participate this study. Exclusion criteria were history of other psychotic disorders, comorbidities with other common medical condition, a history of substance abuse. Sample inspection for serum vitamin D using ELFA method. Statistical analysis using numeric comparative T-independent test. Results: The results showed that average levels of vitamin D for a group of subjects living in areas of high land was 227.6 ng / mL with a standard deviation of 86.78 ng / mL, the lowest levels of vitamin D is 138 ng / mL and the highest 482 ng / mL. In the group of subjects who settled in the low lands seem mean vitamin D levels higher than the mountainous area with an average 237.8 ng / mL with a standard deviation of 100.16 ng / mL. Vitamin D levels are lowest and the highest 138-585 ng / mL. Conclusion and Suggestion: The results of the analysis using the Mann Whitney test showed that there were no significant differences between the mean for the levels of vitamin D based on residence subject with a value of p = 0.652.

Keywords: latitude, schizophrenia, Vitamin D, Sumatera Utara

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