Search results for: cellular degeneration
522 A Low Cost Gain-Coupled Distributed Feedback Laser Based on Periodic Surface p-Contacts
Authors: Yongyi Chen, Li Qin, Peng Jia, Yongqiang Ning, Yun Liu, Lijun Wang
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The distributed feedback (DFB) lasers are indispensable in optical phase array (OPA) used for light detection and ranging (LIDAR) techniques, laser communication systems and integrated optics, thanks to their stable single longitudinal mode and narrow linewidth properties. Traditional index-coupled (IC) DFB lasers with uniform gratings have an inherent problem of lasing two degenerated modes. Phase shifts are usually required to eliminate the mode degeneration, making the grating structure complex and expensive. High-quality antireflection (AR) coatings on both lasing facets are also essential owing to the random facet phases introduced by the chip cleavage process, which means half of the lasing energy is wasted. Gain-coupled DFB (GC-DFB) lasers based on the periodic gain (or loss) are announced to have single longitudinal mode as well as capable of the unsymmetrical coating to increase lasing power and efficiency thanks to facet immunity. However, expensive and time-consuming technologies such as epitaxial regrowth and nanoscale grating processing are still required just as IC-DFB lasers, preventing them from practical applications and commercial markets. In this research, we propose a low-cost, single-mode regrowth-free GC-DFB laser based on periodic surface p-contacts. The gain coupling effect is achieved simply by periodic current distribution in the quantum well caused by periodic surface p-contacts, introducing very little index-coupling effect that can be omitted. It is prepared by i-line lithography, without nanoscale grating fabrication or secondary epitaxy. Due to easy fabrication techniques, it provides a method to fabricate practical low cost GC-DFB lasers for widespread practical applications.Keywords: DFB laser, gain-coupled, low cost, periodic p-contacts
Procedia PDF Downloads 123521 Evaluation of Anti-Typhoid Effects of Azadirachta indica L. Fractions
Authors: A. Adetutu, T. M. Awodugba, O. A. Owoade
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The development of resistance to currently known conventional anti-typhoid drugs has necessitated search into cheap, more potent and less toxic anti-typhoid drugs of plant origin. Therefore, this study investigated the anti-typhoid activity of fractions of A. indica in Salmonella typhi infected rats. Leaves of A. indica were extracted in methanol and fractionated into n-hexane, chloroform, ethyl-acetate, and aqueous fractions. The anti-salmonella potentials of fractions of A. indica were assessed via in-vitro inhibition of S. typhi using agar well diffusion, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and biofilm assays. The biochemical and haematological parameters were determined by spectrophotometric methods. The histological analysis was performed using Haematoxylin and Eosin staining methods. Data analysis was performed by one-way ANOVA. Results of this study showed that S. typhi was sensitive to aqueous and chloroform fractions of A. indica, and the fractions showed biofilm inhibition at concentrations of 12.50, 1.562, and 0.39 mg/mL. In the in-vivo study, the extract and chloroform fraction had significant (p < 0.05) effects on the number of viable S. typhi recovered from the blood and stopped salmonellosis after 6 days of treatment of rats at 500 mg/kg b.w. Treatments of infected rats with chloroform and aqueous fractions of A. indica normalized the haematological parameters in the animals. Similarly, treatment with fractions of the plants sustained a normal antioxidant status when compared with the normal control group. Chloroform and ethyl-acetate fractions of A. indica reversed the liver and intestinal degeneration induced by S. typhi infection in rats. The present investigation indicated that the aqueous and chloroform fractions of A. indica showed the potential to provide an effective treatment for salmonellosis, including typhoid fever. The results of the study may justify the ethno-medicinal use of the extract in traditional medicine for the treatment of typhoid and salmonella infections.Keywords: Azadirachta indica L, salmonella, typhoid, leave fractions
Procedia PDF Downloads 124520 Altered Proteostasis Contributes to Skeletal Muscle Atrophy during Chronic Hypobaric Hypoxia: An Insight into Signaling Mechanisms
Authors: Akanksha Agrawal, Richa Rathor, Geetha Suryakumar
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Muscle represents about ¾ of the body mass, and a healthy muscular system is required for human performance. A healthy muscular system is dynamically balanced via the catabolic and anabolic process. High altitude associated hypoxia altered this redox balance via producing reactive oxygen and nitrogen species that ultimately modulates protein structure and function, hence, disrupts proteostasis or protein homeostasis. The mechanism by which proteostasis is clinched includes regulated protein translation, protein folding, and protein degradation machinery. Perturbation in any of these mechanisms could increase proteome imbalance in the cellular processes. Altered proteostasis in skeletal muscle is likely to be responsible for contributing muscular atrophy in response to hypoxia. Therefore, we planned to elucidate the mechanism involving altered proteostasis leading to skeletal muscle atrophy under chronic hypobaric hypoxia. Material and Methods-Male Sprague Dawley rats weighing about 200-220 were divided into five groups - Control (Normoxic animals), 1d, 3d, 7d and 14d hypobaric hypoxia exposed animals. The animals were exposed to simulated hypoxia equivalent to 282 torr pressure (equivalent to an altitude of 7620m, 8% oxygen) at 25°C. On completion of chronic hypobaric hypoxia (CHH) exposure, rats were sacrificed, muscle was excised and biochemical, histopathological and protein synthesis signaling were studied. Results-A number of changes were observed with the CHH exposure time period. ROS was increased significantly on 07 and 14 days which were attributed to protein oxidation via damaging muscle protein structure by oxidation of amino acids moiety. The oxidative damage to the protein further enhanced the various protein degradation pathways. Calcium activated cysteine proteases and other intracellular proteases participate in protein turnover in muscles. Therefore, we analysed calpain and 20S proteosome activity which were noticeably increased at CHH exposure as compared to control group representing enhanced muscle protein catabolism. Since inflammatory markers (myokines) affect protein synthesis and triggers degradation machinery. So, we determined inflammatory pathway regulated under hypoxic environment. Other striking finding of the study was upregulation of Akt/PKB translational machinery that was increased on CHH exposure. Akt, p-Akt, p70 S6kinase, and GSK- 3β expression were upregulated till 7d of CHH exposure. Apoptosis related markers, caspase-3, caspase-9 and annexin V was also increased on CHH exposure. Conclusion: The present study provides evidence of disrupted proteostasis under chronic hypobaric hypoxia. A profound loss of muscle mass is accompanied by the muscle damage leading to apoptosis and cell death under CHH. These cellular stress response pathways may play a pivotal role in hypobaric hypoxia induced skeletal muscle atrophy. Further research in these signaling pathways will lead to development of therapeutic interventions for amelioration of hypoxia induced muscle atrophy.Keywords: Akt/PKB translational machinery, chronic hypobaric hypoxia, muscle atrophy, protein degradation
Procedia PDF Downloads 265519 High-Intensity, Short-Duration Electric Pulses Induced Action Potential in Animal Nerves
Authors: Jiahui Song, Ravindra P. Joshi
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The use of high-intensity, short-duration electric pulses is a promising development with many biomedical applications. The uses include irreversible electroporation for killing abnormal cells, reversible poration for drug and gene delivery, neuromuscular manipulation, and the shrinkage of tumors, etc. High intensity, short-duration electric pulses result in the creation of high-density, nanometer-sized pores in the cellular membrane. This electroporation amounts to localized modulation of the transverse membrane conductance, and effectively provides a voltage shunt. The electrically controlled changes in the trans-membrane conductivity could be used to affect neural traffic and action potential propagation. A rat was taken as the representative example in this research. The simulation study shows the pathway from the sensorimotor cortex down to the spinal motoneurons, and effector muscles could be reversibly blocked by using high-intensity, short-duration electrical pulses. Also, actual experimental observations were compared against simulation predictions.Keywords: action potential, electroporation, high-intensity, short-duration
Procedia PDF Downloads 262518 Linkage between a Plant-based Diet and Visual Impairment: A Systematic Review and Meta-Analysis
Authors: Cristina Cirone, Katrina Cirone, Monali S. Malvankar-Mehta
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Purpose: An increased risk of visual impairment has been observed in individuals lacking a balanced diet. The purpose of this paper is to characterize the relationship between plant-based diets and specific ocular outcomes among adults. Design: Systematic review and meta-analysis. Methods: This systematic review and meta-analysis were conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement guidelines. The databases MEDLINE, EMBASE, Cochrane, and PubMed, were systematically searched up until May 27, 2021. Of the 503 articles independently screened by two reviewers, 21 were included in this review. Quality assessment and data extraction were performed by both reviewers. Meta-analysis was conducted using STATA 15.0. Fixed-effect and random-effect models were computed based on heterogeneity. Results: A total of 503 studies were identified which then underwent duplicate removal and a title and abstract screen. The remaining 61 studies underwent a full-text screen, 21 progressed to data extraction and fifteen were included in the quantitative analysis. Meta-analysis indicated that regular consumption of fish (OR = 0.70; CI: [0.62-0.79]) and skim milk, poultry, and non-meat animal products (OR = 0.70; CI: [0.61-0.79]) is positively correlated with a reduced risk of visual impairment (age-related macular degeneration, age-related maculopathy, cataract development, and central geographic atrophy) among adults. Consumption of red meat [OR = 1.41; CI: [1.07-1.86]) is associated with an increased risk of visual impairment. Conclusion: Overall, a pescatarian diet is associated with the most favorable visual outcomes among adults, while the consumption of red meat appears to negatively impact vision. Results suggest a need for more local and government-led interventions promoting a healthy and balanced diet.Keywords: plant-based diet, pescatarian diet, visual impairment, systematic review, meta-analysis
Procedia PDF Downloads 177517 Effect of Insulin versus Green Tea on the Parotid Gland of Streptozotocin Induced Diabetic Rats
Authors: H. El-Messiry, M. El-Zainy, D. Ghazy
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Diabetes is a metabolic disease that results in a variety of oral health complications. Green tea is a natural antioxidant proved to have powerful effects against diabetes. The aim of this study was to compare between the effect of insulin and green tea on the Parotid gland of streptozotocin induced diabetic Albino rats by using light and transmission electron microscopy. Forty male Albino rats were divided into control group and diabetic groups. The diabetic group received a single injection of 40 mg/kg of streptozotocin intra-peritoneal under anesthesia and was further subdivided into three subgroups: The diabetic untreated subgroup which was untreated for two weeks, the insulin treated subgroup which has received insulin subcutaneously in a daily dose of 5 IU/kg body weight/day for two weeks and a green tea treated subgroup received a daily dose of 1 ml/ 100 gm body weight intragastrically for two weeks. Rats were terminated and parotid glands were dissected and processed for light and transmission electron microscopic examination. Histological examination of the diabetic untreated subgroup revealed acinar cells with pyknotic and hyperchromatic nuclei with cytoplasmic vacuolations. Ultrastructurally, acinar cells showed nuclear pleomorphism, dilated rough endoplasmic reticulum and swollen mitochondria with damaged cristae. Inflammatory cell infiltration was detected both histologically and ultrastructurally. Ducts showed signs of degeneration with loss of their normal outline and stagnated secretion within the lumen. However, insulin and green tea treated subgroups showed minimal degenerative damage and were almost similar to the control with minimal changes. Treatment of the parotid gland of the streptozotocin induced diabetic rats with GT was closely comparable to the traditional insulin therapy in reducing signs of histological and ultrastructural damage.Keywords: diabetes, green tea, insulin, parotid
Procedia PDF Downloads 176516 Coverage Probability Analysis of WiMAX Network under Additive White Gaussian Noise and Predicted Empirical Path Loss Model
Authors: Chaudhuri Manoj Kumar Swain, Susmita Das
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This paper explores a detailed procedure of predicting a path loss (PL) model and its application in estimating the coverage probability in a WiMAX network. For this a hybrid approach is followed in predicting an empirical PL model of a 2.65 GHz WiMAX network deployed in a suburban environment. Data collection, statistical analysis, and regression analysis are the phases of operations incorporated in this approach and the importance of each of these phases has been discussed properly. The procedure of collecting data such as received signal strength indicator (RSSI) through experimental set up is demonstrated. From the collected data set, empirical PL and RSSI models are predicted with regression technique. Furthermore, with the aid of the predicted PL model, essential parameters such as PL exponent as well as the coverage probability of the network are evaluated. This research work may assist in the process of deployment and optimisation of any cellular network significantly.Keywords: WiMAX, RSSI, path loss, coverage probability, regression analysis
Procedia PDF Downloads 164515 Experimental Investigation on the Efficiency of Expanded Polystyrene Geofoam Post and Beam System in Protecting Lifelines
Authors: Masood Abdollahi, Seyed Naser Moghaddas Tafreshi
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Expanded polystyrene (EPS) geofoam is a cellular geosynthetic material that can be used to protect lifelines (e.g. pipelines, electricity cables, etc.) below ground. Post and beam system is the most recent configuration of EPS blocks which can be implemented for this purpose. It provides a void space atop lifelines which allows settlement of the loading surface with imposing no pressure on the lifelines system. This paper investigates the efficiency of the configuration of post-beam system subjected to static loading. To evaluate the soil surface settlement, beam deformation and transferred pressure over the beam, laboratory tests using two different densities for EPS blocks are conducted. The effect of geogrid-reinforcing the cover soil on system response is also investigated. The experimental results show favorable performance of EPS post and beam configuration in protecting underground lifelines.Keywords: beam deformation, EPS block, laboratory test, post-Beam system, soil surface settlement
Procedia PDF Downloads 235514 Mobile Microscope for the Detection of Pathogenic Cells Using Image Processing
Authors: P. S. Surya Meghana, K. Lingeshwaran, C. Kannan, V. Raghavendran, C. Priya
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One of the most basic and powerful tools in all of science and medicine is the light microscope, the fundamental device for laboratory as well as research purposes. With the improving technology, the need for portable, economic and user-friendly instruments is in high demand. The conventional microscope fails to live up to the emerging trend. Also, adequate access to healthcare is not widely available, especially in developing countries. The most basic step towards the curing of a malady is the diagnosis of the disease itself. The main aim of this paper is to diagnose Malaria with the most common device, cell phones, which prove to be the immediate solution for most of the modern day needs with the development of wireless infrastructure allowing to compute and communicate on the move. This opened up the opportunity to develop novel imaging, sensing, and diagnostics platforms using mobile phones as an underlying platform to address the global demand for accurate, sensitive, cost-effective, and field-portable measurement devices for use in remote and resource-limited settings around the world.Keywords: cellular, hand-held, health care, image processing, malarial parasites, microscope
Procedia PDF Downloads 261513 Biosynthesis of Titanium Dioxide Nanoparticles and Their Antibacterial Property
Authors: Prachi Singh
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This paper presents a low-cost, eco-friendly and reproducible microbe mediated biosynthesis of TiO2 nanoparticles. TiO2 nanoparticles synthesized using the bacterium, Bacillus subtilis, from titanium as a precursor, were confirmed by TEM analysis. The morphological characteristics state spherical shape, with the size of individual or aggregate nanoparticles, around 30-40 nm. Microbial resistance represents a challenge for the scientific community to develop new bioactive compounds. Here, the antibacterial effect of TiO2 nanoparticles on Escherichia coli was investigated, which was confirmed by CFU (Colony-forming unit). Further, growth curve study of E. coli Hb101 in the presence and absence of TiO2 nanoparticles was done. Optical density decrease was observed with the increase in the concentration of TiO2. It could be attributed to the inactivation of cellular enzymes and DNA by binding to electron-donating groups such as carboxylates, amides, indoles, hydroxyls, thiols, etc. which cause little pores in bacterial cell walls, leading to increased permeability and cell death. This justifies that TiO2 nanoparticles have efficient antibacterial effect and have potential to be used as an antibacterial agent for different purposes.Keywords: antibacterial effect, CFU, Escherichia coli Hb101, growth curve, TEM, TiO2 nanoparticle, Toxicity, UV-Vis
Procedia PDF Downloads 290512 Identification and Characterization of Small Peptides Encoded by Small Open Reading Frames using Mass Spectrometry and Bioinformatics
Authors: Su Mon Saw, Joe Rothnagel
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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs
Procedia PDF Downloads 184511 Presence, Distribution and Form of Calcium Oxalate Crystals in Relation to Age of Actinidia Deliciosa Leaves and Petioles
Authors: Muccifora S., Rinallo C., Bellani L.
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Calcium (Ca²+) is an element essential to the plant being involved in plant growth and development. At high concentrations, it is toxic and can influence every stage, process and cellular activity of plant life. Given its toxicity, cells implement mechanisms to compartmentalize calcium in a vacuole, endoplasmic reticulum, mitochondria, plastids and cell wall. One of the most effective mechanisms to reduce the excess of calcium, thus avoiding cellular damage, is its complexation with oxalic acid to form calcium oxalate crystals that are no longer osmotically or physiologically active. However, the sequestered calcium can be mobilized when the plant needs it. Calcium crystals can be accumulated in the vacuole of specialized sink-cells called idioblasts, with different crystalline forms (druse, raphyde and styloid) of diverse physiological meanings. Actinidia deliciosa cv. Hayward presents raphydes and styloid localized in idioblasts in cells of photosynthetic and non-photosynthetic tissues. The purpose of this work was to understand if there is a relationship between the age of Actinidia leaves and the presence, distribution, dimension and shape of oxalate crystals by means of light, fluorescent, polarized and transmission electron microscopy. Three vines from female plants were chosen at the beginning of the season and used throughout the study. The leaves with petioles were collected at various stages of development from the bottom to the shoot of the plants monthly from April to July. The samples were taken in corresponding areas of the central and lateral parts of the leaves and of the basal portion of the petiole. The results showed that in the leaves, the number of raphyde idioblasts decreased with the progress of the growing season, while the styloid idioblasts increased progressively, becoming very numerous in the upper nodes of July. In June and in July samples, in the vacuoles of the highest nodes, a portion regular in shape strongly stained with rubeanic acid was present. Moreover, the chlortetracycline (CTC) staining for localization of free calcium marked the wall of the idioblasts and the wall of the cells near vascular bundles. In April petiole samples, moving towards the youngest nodes, the raphydes idioblast decreased in number and in the length of the single raphydes. Besides, crystals stained with rubeanic acid appeared in the vacuoles of some cells. In June samples, numerous raphyde idioblasts oriented parallel to vascular bundles were evident. Under the electron microscope, numerous idioblasts presented not homogeneous electrondense aggregates of material, in which a few crystals (styloids) in the form of regular holes were scattered. In July samples, an increase in the number of styloid idioblasts in the youngest nodes and little masses stained with CTC near styloids were observed. Peculiar cells stained with rubeanic acid were detected and hypothesized to be involved in the formation of the idioblasts. In conclusion, in Actinidia leaves and petioles, it seems to confirm the hypothesis that the formation of styloid idioblasts can be correlated to increasing calcium levels in growing tissues.Keywords: calcium oxalate crystals, actinidia deliciosa, light and electron microscopy, idioblasts
Procedia PDF Downloads 78510 The Expression of a Novel Gene Encoding an Ankyrin-Repeat Protein, DRA1, Is Regulated by Drought-Responsive Alternative Splicing
Authors: H. Sakamoto, Y. Nakagawara, S. Oguri
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Drought stress is a critical environmental factor that adversely affects crop productivity and quality. Because of their immobile nature, plants have evolved mechanisms to sense and respond to drought stress. We identified a novel locus of Arabidopsis, designated DRA1 (drought responsive ankyrin 1), whose disruption leads to increased drought stress tolerance. DRA1 encodes a transmembrane protein with an ankyrin repeat motif that has been implicated in diverse cellular processes such as signal transduction. RT-PCR analysis revealed that there were at least two splicing variants of DRA1 transcripts in wild type plants. In response to drought stress, the levels of DRA1 transcripts retaining second and third introns were increased, whereas these introns were removed under unstressed conditions. These results suggest that DRA1 protein may negatively regulate plant drought tolerance and that the expression of DRA1 is regulated in response to drought stress by alternative splicing.Keywords: alternative splicing, ankyrin repeat, Arabidopsis, drought tolerance
Procedia PDF Downloads 316509 Prophylactic and Curative Effect of Selenium on Infertility Induced by Formaldehyde Using Male Albino Mice
Authors: Suhera M. Aburawi, Habiba A. El Jaafari, Soad A. Treesh, Abdulssalam M. Abu-Aisha, Faisal S. Alwaer, Reda A. Eltubuly, Medeha Elghedamsi
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Introduction: Infertility is a source of psychological, and sometimes social, stress on parents who desire to have children. Formaldehyde is used chiefly as disinfectant, preservative and in the chemical synthesis. The medical uses of formaldehyde are limited, but focused especially on laboratory use. Selenium is an essential trace mineral element for human; it is essential for sperm function and male fertility. Selenium deficiency has been linked to reproductive problems in animals. Objectives: To investigate the prophylactic and curative effect of selenium on male infertility induced by formaldehyde using male albino mice. Method: Forty male albino mice were used, weight 25-30 gm. Five groups of male mice (n=8) were used. Group 1 was daily administered water for injection (5ml/kg) for five days, group 2 was daily administered selenium (100 μg/kg) for five days, group 3 was daily administered formaldehyde (30mg/kg) for five days, group 4 (prophylaxis) was daily administered a combination of formaldehyde and selenium for five days, while group 5 (curative) was daily administered formaldehyde for five days followed by daily administration of selenium for the next five days. Intraperitoneal administration was adopted. At the end of the administration, seminal fluid was collected from vas deferens. Sperm count, morphology and motility were scored; histopathological screening of genital system was carried out. SPSS was applied for comparing groups. Results and conclusion: It was found that formaldehyde toxicity did not change the sperm count and percentage of motile sperm; unhealthy sperm was increased, while healthy sperm was decreased. Formaldehyde produces degeneration/damage to the male mice genital system. Selenium alone produce an increase in sperm count, volume of seminal fluid and the percentage of motile sperm. Selenium has prophylactic and curative effects against formaldehyde-induce genital system toxicity. Future work is recommended to find out if selenium protective effect is through antioxidant or other mechanisms.Keywords: infertility, formaldehyde, selenium, male mice
Procedia PDF Downloads 418508 Defective Autophagy Disturbs Neural Migration and Network Activity in hiPSC-Derived Cockayne Syndrome B Disease Models
Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche
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It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells
Procedia PDF Downloads 117507 Effects of Stiffness on Endothelial Cells Behavior
Authors: Forough Ataollahi, Sumit Pramanik, Belinda Pingguan-Murphy, Wan Abu Bakar Bin Wan Abas, Noor Azuan Bin Abu Osman
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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al2O3) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al2O3 at curing temperature 50˚C for 4 h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.Keywords: stiffness, proliferation, bovine aortic endothelial cells, extra cellular matrix, vascular
Procedia PDF Downloads 336506 Protective Effect of Probiotic Lactic Acid Bacteria on Thioacetamide-Induced Liver Fibrosis in Rats: Histomorphological Study
Authors: Chittapon Jantararussamee, Malai Taweechotipatr, Udomsri Showpittapornchai, Wisuit Pradidarcheep
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Hepatic fibrosis is characterized by collagen accumulation in hepatic lobules following wound healing process. If lefts untreated, it could progress into hepatic cirrhosis, portal hypertension, and liver failure. Probiotics comprise of lactic acid bacteria which are crucial components of the intestinal microflora and possess many beneficial properties. The objective of this study is to investigate the hepatoprotective effects of probiotic lactic acid bacteria (mixture of Lactobacillus paracasei, Lactobacillus casei, and Lactobacillus confusus at a ratio of 1: 1: 1) on thioacetamide-induced liver fibrotic rats in term of histomorphology study. Twenty-four male Wistar rats were randomly divided into four groups with 6 rats each: (A) control, (B) fibrotic, (C) fibrotic+probiotic, and (D) probiotic. Group (A) received daily oral administration of distilled water. Group (B and C) were induced by intraperitoneal injection of thioacetamide (TAA) (200 mg/kg BW) 3 times per week for consecutive 8 weeks. In probiotic-treated group (C and D), the number of a mixture of the viable microbial cells at 10⁹ CFU/ml was administered orally daily. After sacrifice, liver tissues were collected and processed for routine histological technique and stained with Sirius red. It was found that the fibrotic rats showed hepatic injury marked by area of inflammation, hydropic degeneration of hepatocytes, and accumulation of myofibroblast-like cells. The collagen fibers were substantially accumulated in the hepatic lobules. Moreover, probiotic-treated group significantly reduced the accumulation of collagen in rats treated by TAA. The liver damage was found to be lesser in the probiotic-treated group. It was noted that the liver tissues of control and probiotics groups were shown to be normal. Administration with probiotic lactic acid bacteria could improve the histomorphology in fibrotic liver and be useful for prevention of hepatic disorders.Keywords: liver fibrosis, probiotics, lactic acid bacteria, thioacetamide
Procedia PDF Downloads 123505 Biochemical Changes in the Liver of Mice after Exposure to Different Doses of Diclofenac Sodium
Authors: Deepak Mohan, Sushma Sharma
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Non-Steroidal Anti-Inflammatory Drugs (NSAIDs) are a group of widely used drugs for the treatment of rheumatoid diseases and to relieve pain and inflammation due to their analgesic anti-pyretic and anti-inflammatory properties. The therapeutic and many of the toxic effects of NSAIDs result from reversible inhibition of enzymes in the cyclooxygenase (COX) group. In the present investigation the effect of the drug on the concentration of lipids, and on the activity of the enzymes i.e. acid and alkaline phosphatase, GOT, GPT and lipid peroxidase were studied. There was a significant enhancement in the activities of both acid and alkaline phosphatase after 21 days of treatment. Proportionate increase in the MDA contents was observed after different days of diclofenac treatment. Cellular damage in the liver resulted in decrease in the activity of both GOT (Glutamate oxaloacetate transaminase) and GPT (Glutamate pyruvate transaminase) in both low and high dose groups. Significant decrease in the liver contents was also observed in both dose groups.Keywords: anti-inflammatory, cyclooxygenase, glutamate oxaloacetate transaminase, malondialdehyde
Procedia PDF Downloads 294504 Induction of Hsp70 and Antioxidant Status in Porcine Granulosa Cells in Response to Deoxynivalenol and Zearalenone Exposure in vitro
Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin
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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.Keywords: deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells
Procedia PDF Downloads 247503 Performance Evaluation of a Minimum Mean Square Error-Based Physical Sidelink Share Channel Receiver under Fading Channel
Authors: Yang Fu, Jaime Rodrigo Navarro, Jose F. Monserrat, Faiza Bouchmal, Oscar Carrasco Quilis
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Cellular Vehicle to Everything (C-V2X) is considered a promising solution for future autonomous driving. From Release 16 to Release 17, the Third Generation Partnership Project (3GPP) has introduced the definitions and services for 5G New Radio (NR) V2X. Experience from previous generations has shown that establishing a simulator for C-V2X communications is an essential preliminary step to achieve reliable and stable communication links. This paper proposes a complete framework of a link-level simulator based on the 3GPP specifications for the Physical Sidelink Share Channel (PSSCH) of the 5G NR Physical Layer (PHY). In this framework, several algorithms in the receiver part, i.e., sliding window in channel estimation and Minimum Mean Square Error (MMSE)-based equalization, are developed. Finally, the performance of the developed PSSCH receiver is validated through extensive simulations under different assumptions.Keywords: C-V2X, channel estimation, link-level simulator, sidelink, 3GPP
Procedia PDF Downloads 115502 Evaluation of Wound Healing Activity of Curcuma purpurascens BI. Rhizomes in Rats
Authors: Elham Rouhollahi, Soheil Zorofchian Moghadamtousi, Salma Baig, Mahmood Ameen Abdulla, Zahurin Mohamed
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This study was designed to assess cutaneous wound healing potential of hexane extract of Curcuma purpurascens rhizomes (HECP). Twenty-four rats were divided into 4 groups: 1. Negative, 2. Low dose, 3. High dose and 4. Treatment, with 6 rats in each group. Full-thickness incisions with a diameter of 2 cm were made on the back of each rat. Rats were topically treated two times a day for 15 days. Group 1-4 were treated with sterile distilled water, 5% and 10% of extract and intrasite gel, respectively. Masson's trichrome and hematoxylin staining techniques are employed for histological analysis revealed strong wound healing potential closer to that of conventional drug intrasite gel. HECP significantly decreased wound area and an increase in hydroxyproline, cellular proliferation, the number of blood vessels and the level of collagen synthesis was observed. Thus, it could be concluded that HECP possesses strong wound healing potential.Keywords: Curcuma purpurascens, wound healing, histopathology, hematoxylin staining
Procedia PDF Downloads 428501 Indigenous Patch Clamp Technique: Design of Highly Sensitive Amplifier Circuit for Measuring and Monitoring of Real Time Ultra Low Ionic Current through Cellular Gates
Authors: Moez ul Hassan, Bushra Noman, Sarmad Hameed, Shahab Mehmood, Asma Bashir
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The importance of Noble prize winning “Patch Clamp Technique” is well documented. However, Patch Clamp Technique is very expensive and hence hinders research in developing countries. In this paper, detection, processing and recording of ultra low current from induced cells by using transimpedence amplifier is described. The sensitivity of the proposed amplifier is in the range of femto amperes (fA). Capacitive-feedback is used with active load to obtain a 20MΩ transimpedance gain. The challenging task in designing includes achieving adequate performance in gain, noise immunity and stability. The circuit designed by the authors was able to measure current in the rangeof 300fA to 100pA. Adequate performance shown by the amplifier with different input current and outcome result was found to be within the acceptable error range. Results were recorded using LabVIEW 8.5®for further research.Keywords: drug discovery, ionic current, operational amplifier, patch clamp
Procedia PDF Downloads 512500 Structural Correlates of Reduced Malicious Pleasure in Huntington's Disease
Authors: Sandra Baez, Mariana Pino, Mildred Berrio, Hernando Santamaria-Garcia, Lucas Sedeno, Adolfo Garcia, Sol Fittipaldi, Agustin Ibanez
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Schadenfreude refers to the perceiver’s experience of pleasure at another’s misfortune. This is a multidetermined emotion which can be evoked by hostile feelings and envy. The experience of Schadenfreude engages mechanisms implicated in diverse social cognitive processes. For instance, Schadenfreude involves heightened reward processing, accompanied by increased striatal engagement and it interacts with mentalizing and perspective-taking abilities. Patients with Huntington's disease (HD) exhibit reductions of Schadenfreude experience, suggesting a role of striatal degeneration in such an impairment. However, no study has directly assessed the relationship between regional brain atrophy in HD and reduced Schadenfreude. This study investigated whether gray matter (GM) atrophy in HD patients correlates with ratings of Schadenfreude. First, we compared the performance of 20 HD patients and 23 controls on an experimental task designed to trigger Schadenfreude and envy (another social emotion acting as a control condition). Second, we compared GM volume between groups. Third, we examined brain regions where atrophy might be associated with specific impairments in the patients. Results showed that while both groups showed similar ratings of envy, HD patients reported lower Schadenfreude. The latter pattern was related to atrophy in regions of the reward system (ventral striatum) and the mentalizing network (precuneus and superior parietal lobule). Our results shed light on the intertwining of reward and socioemotional processes in Schadenfreude, while offering novel evidence about their neural correlates. In addition, our results open the door to future studies investigating social emotion processing in other clinical populations characterized by striatal or mentalizing network impairments (e.g., Parkinson’s disease, schizophrenia, autism spectrum disorders).Keywords: envy, Gray matter atrophy, Huntigton's disease, Schadenfreude, social emotions
Procedia PDF Downloads 329499 Gene Expressions in Left Ventricle Heart Tissue of Rat after 150 Mev Proton Irradiation
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Introduction: In mediastinal radiotherapy and to a lesser extend also in total-body irradiation (TBI) radiation exposure may lead to development of cardiac diseases. Radiation-induced heart disease is dose-dependent and it is characterized by a loss of cardiac function, associated with progressive heart cells degeneration. We aimed to determine the in-vivo radiation effects on fibronectin, ColaA1, ColaA2, galectin and TGFb1 gene expression levels in left ventricle heart tissues of rats after irradiation. Material and method: Four non-treatment adult Wistar rats as control group (group A) were selected. In group B, 4 adult Wistar rats irradiated to 20 Gy single dose of 150 Mev proton beam locally in heart only. In heart plus lung irradiate group (group C) 4 adult rats was irradiated by 50% of lung laterally plus heart radiation that mentioned in before group. At 8 weeks after radiation animals sacrificed and left ventricle heart dropped in liquid nitrogen for RNA extraction by Absolutely RNA® Miniprep Kit (Stratagen, Cat no. 400800). cDNA was synthesized using M-MLV reverse transcriptase (Life Technologies, Cat no. 28025-013). We used Bio-Rad machine (Bio Rad iQ5 Real Time PCR) for QPCR testing by relative standard curve method. Results: We found that gene expression of fibronectin in group C significantly increased compared to control group, but it was not showed significant change in group B compared to group A. The levels of gene expressions of Cola1 and Cola2 in mRNA did not show any significant changes between normal and radiation groups. Changes of expression of galectin target significantly increased only in group C compared to group A. TGFb1 expressions in group C more than group B showed significant enhancement compared to group A. Conclusion: In summary we can say that 20 Gy of proton exposure of heart tissue may lead to detectable damages in heart cells and may distribute function of them as a component of heart tissue structure in molecular level.Keywords: gene expression, heart damage, proton irradiation, radiotherapy
Procedia PDF Downloads 484498 An Activatable Prodrug for the Treatment of Metastatic Tumors
Authors: Eun-Joong Kim, Sankarprasad Bhuniya, Hyunseung Lee, Hyun Min Kim, Chaejoon Cheong, Su-khendu Maiti, Kwan Soo Hong, Jong Seung Kim
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Metastatic cancers have historically been difficult to treat. However, metastatic tumors have been found to have high levels of reactive oxygen species such as hydrogen peroxide (H2O2), supporting the hypothesis that a prodrug could be activated by intracellular H2O2 and lead to a potential anti-metastatic therapy. In this study, prodrug 7 was designed to be activated by H2O2-mediated boronate oxidation, resulting in activation of the fluorophore for detection and release of the therapeutic agent, SN-38. Drug release from prodrug 7 was investigated by monitoring fluorescence after addition of H2O2 to the cancer cells. Prodrug 7 activated by H2O2 selectively inhibited tumor cell growth. Furthermore, intratracheally administered prodrug 7 showed effective anti-tumor activity in a mouse model of metastatic lung disease. Thus, this H2O2-responsive prodrug has therapeutic potential as a novel treatment for metastatic cancer via cellular imaging with fluorescence as well as selective release of the anti-cancer drug, SN-38.Keywords: hydrogen peroxide, prodrug, metastatic tumors, fluorescence
Procedia PDF Downloads 450497 Influence of the Refractory Period on Neural Networks Based on the Recognition of Neural Signatures
Authors: José Luis Carrillo-Medina, Roberto Latorre
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Experimental evidence has revealed that different living neural systems can sign their output signals with some specific neural signature. Although experimental and modeling results suggest that neural signatures can have an important role in the activity of neural networks in order to identify the source of the information or to contextualize a message, the functional meaning of these neural fingerprints is still unclear. The existence of cellular mechanisms to identify the origin of individual neural signals can be a powerful information processing strategy for the nervous system. We have recently built different models to study the ability of a neural network to process information based on the emission and recognition of specific neural fingerprints. In this paper we further analyze the features that can influence on the information processing ability of this kind of networks. In particular, we focus on the role that the duration of a refractory period in each neuron after emitting a signed message can play in the network collective dynamics.Keywords: neural signature, neural fingerprint, processing based on signal identification, self-organizing neural network
Procedia PDF Downloads 484496 Understanding the Thermal Resistance of Active Dry Yeast by Differential Scanning Calorimetry Approach
Authors: Pauline Ribert, Gaelle Roudaut, Sebastien Dupont, Laurent Beney
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Yeasts, anhydrobiotic organisms, can survive extreme water disturbances, thanks to the prolonged and reversible suspension of their cellular activity as well as the establishment of a defense arsenal. This property is exploited by many industrialists. One of the protection systems implemented by yeast is the vitrification of its cytoplasm by trehalose. The thermal resistance of dry yeasts is a crucial parameter for their use. However, studies on the thermal resistance of dry yeasts are often based on yeasts produced in laboratory conditions with non-optimal drying processes. We, therefore, propose a study on the thermal resistance of industrial dry yeasts in relation to their thermophysical properties. Heat stress was applied at three temperatures (50, 75, and 100°C) for 10, 30, or 60-minute treatments. The survival of yeasts to these treatments was estimated, and their thermophysical properties were studied by differential scanning calorimetry. The industrial dry yeasts resisted 60 minutes at 50°C and 75°C and 10 minutes at a temperature close to 100°C. At 100°C, yeast was above their glass transition temperature. Industrial dry yeasts are therefore capable of withstanding high thermal stress if maintained in a specific thermophysical state.Keywords: dry yeast, glass transition, thermal resistance, vitrification
Procedia PDF Downloads 146495 Constraint-Based Computational Modelling of Bioenergetic Pathway Switching in Synaptic Mitochondria from Parkinson's Disease Patients
Authors: Diana C. El Assal, Fatima Monteiro, Caroline May, Peter Barbuti, Silvia Bolognin, Averina Nicolae, Hulda Haraldsdottir, Lemmer R. P. El Assal, Swagatika Sahoo, Longfei Mao, Jens Schwamborn, Rejko Kruger, Ines Thiele, Kathrin Marcus, Ronan M. T. Fleming
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Degeneration of substantia nigra pars compacta dopaminergic neurons is one of the hallmarks of Parkinson's disease. These neurons have a highly complex axonal arborisation and a high energy demand, so any reduction in ATP synthesis could lead to an imbalance between supply and demand, thereby impeding normal neuronal bioenergetic requirements. Synaptic mitochondria exhibit increased vulnerability to dysfunction in Parkinson's disease. After biogenesis in and transport from the cell body, synaptic mitochondria become highly dependent upon oxidative phosphorylation. We applied a systems biochemistry approach to identify the metabolic pathways used by neuronal mitochondria for energy generation. The mitochondrial component of an existing manual reconstruction of human metabolism was extended with manual curation of the biochemical literature and specialised using omics data from Parkinson's disease patients and controls, to generate reconstructions of synaptic and somal mitochondrial metabolism. These reconstructions were converted into stoichiometrically- and fluxconsistent constraint-based computational models. These models predict that Parkinson's disease is accompanied by an increase in the rate of glycolysis and a decrease in the rate of oxidative phosphorylation within synaptic mitochondria. This is consistent with independent experimental reports of a compensatory switching of bioenergetic pathways in the putamen of post-mortem Parkinson's disease patients. Ongoing work, in the context of the SysMedPD project is aimed at computational prediction of mitochondrial drug targets to slow the progression of neurodegeneration in the subset of Parkinson's disease patients with overt mitochondrial dysfunction.Keywords: bioenergetics, mitochondria, Parkinson's disease, systems biochemistry
Procedia PDF Downloads 288494 The Effect of Olea europea L. Extract on Doxorubicin-Induced Cardiotoxicity
Authors: Jessica Maiuolo, Irene Bava, Micaela Gliozzi, Vincenzo Mollace
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Doxorubicin is an anthracycline that is commonly used as a chemotherapy drug due to its cytotoxic effects. The clinical use of doxorubicin is limited due to its known cardiotoxic effects. Polyphenols have a wide range of beneficial properties, and particular importance is given to Oleuropein, one of the main polyphenolic compounds of olive oil. The biological mechanisms involved and the role of the endoplasmic reticulum were examined. Olive oil extract and Oleuropein were able to decrease the damage induced by exposure to doxorubicin. In particular, this natural compound was found to reduce cell mortality and oxidative damage, increase lipid content, and decrease the concentration of calcium ions that escaped from the endoplasmic reticulum. In addition, the direct involvement of this cellular organelle was demonstrated by silencing the ATF6 arm of the Unfolded Protein Response, which was activated after treatment with doxorubicin. The protection afforded by pre-treatment with the natural compound of interest, following the early damage induced by DOXO, provided valuable information regarding the potential use of these substances along with chemotherapy treatment.Keywords: Olea europea L., oleuropein, doxorubicin, endoplasmic reticulum, nutraceutical support
Procedia PDF Downloads 106493 Improved Predictive Models for the IRMA Network Using Nonlinear Optimisation
Authors: Vishwesh Kulkarni, Nikhil Bellarykar
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Cellular complexity stems from the interactions among thousands of different molecular species. Thanks to the emerging fields of systems and synthetic biology, scientists are beginning to unravel these regulatory, signaling, and metabolic interactions and to understand their coordinated action. Reverse engineering of biological networks has has several benefits but a poor quality of data combined with the difficulty in reproducing it limits the applicability of these methods. A few years back, many of the commonly used predictive algorithms were tested on a network constructed in the yeast Saccharomyces cerevisiae (S. cerevisiae) to resolve this issue. The network was a synthetic network of five genes regulating each other for the so-called in vivo reverse-engineering and modeling assessment (IRMA). The network was constructed in S. cereviase since it is a simple and well characterized organism. The synthetic network included a variety of regulatory interactions, thus capturing the behaviour of larger eukaryotic gene networks on a smaller scale. We derive a new set of algorithms by solving a nonlinear optimization problem and show how these algorithms outperform other algorithms on these datasets.Keywords: synthetic gene network, network identification, optimization, nonlinear modeling
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