Search results for: gene regulation
804 The Effect of the Epstein-Barr Virus on the Development of Multiple Sclerosis
Authors: Sina Mahdavi
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Background and Objective: Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the central nervous system (CNS) that affects the myelination process in the CNS. Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially Epstein-Barr virus (EBV) and MS, is one potential cause that is not well understood. In this study, we aim to summarize the available data on EBV infection in MS disease progression. Materials and Methods: For this study, the keywords "Multiple sclerosis," "Epstein-Barr virus," and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched, and 14 articles were chosen, studied, and analyzed. Results: Demyelinated lesions isolated from MS patients contain EBNAs from EBV proteins. The EBNA1 domain contains a pentapeptide fragment identical to B-crystallin, a heat shock peptide, that is increased in peripheral B cells in response to B-crystallin infection, resulting in myelin-directed autoimmunity mediated by proinflammatory T cells. EBNA2, which is involved in the regulation of viral transcription, may enhance transcription from MS risk loci. A 7-fold increase in the risk of MS has been observed in EBV infection with HLA-DR15 synergy. Conclusion: EBV infection along with a variety of specific genetic risk alleles, cause inflammatory cascades in the CNS by infected B cells. There is a high expression of EBV during the course of MS, which indicates the relationship between EBV and MS, that this virus can play a role in the development of MS by creating an inflammatory state. Therefore, measures to modulate the expression of EBV may be effective in reducing inflammatory processes in demyelinated areas of MS patients.Keywords: multiple sclerosis, Epstein-Barr virus, central nervous system, EBNAs
Procedia PDF Downloads 99803 Frequency Control of Self-Excited Induction Generator Based Microgrid during Transition from Grid Connected to Island Mode
Authors: Azhar Ulhaq, Zubair Yameen, Almas Anjum
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Frequency behaviour of self-excited induction generator (SEIG) wind turbines during control mode transition from grid connected to islanded mode is studied in detail. A robust control scheme for frequency regulation based on combined action of STATCOM, energy storage system (ESS) and pitch angle control for wind powered microgrid (MG) is proposed. Suggested STATCOM controller comprises a 3-phase voltage source converter (VSC) that contains insulated gate bipolar transistors (IGBTs) based pulse width modulation (PWM) inverters along with a capacitor bank. Energy storage system control consists of current controlled voltage source converter and battery bank. Both of them acting simultaneously after detection of island compensates for reactive and active power demands, thus regulating frequency at point of common coupling (PCC) and also improves load stability. STATCOM integrates at point of common coupling and ESS is connected to microgrids main bus. Results reveal that proposed control not only stabilizes frequency during transition duration but also minimizes sudden frequency imbalance caused by load variation or wind intermittencies in islanded operation. System is investigated with and without suggested control scheme. The efficacy of proposed strategy has been verified by simulation in MATLAB/Simulink.Keywords: energy storage system, island, wind, STATCOM, self-excited induction generator, SEIG, transient
Procedia PDF Downloads 158802 Molecular Epidemiology of Circulating Adenovirus Types in Acute Conjunctivitis Cases in Chandigarh, North India
Authors: Mini P. Singh, Jagat Ram, Archit Kumar, Tripti Rungta, Jasmine Khurana, Amit Gupta, R. K. Ratho
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Introduction: Human adenovirus is the most common agent involved in viral conjunctivitis. The clinical manifestations vary with different serotypes. The identification of the circulating strains followed by phylogenetic analysis can be helpful in understanding the origin and transmission of the disease. The present study aimed to carry out molecular epidemiology of the adenovirus types in the patients with conjunctivitis presenting to the eye centre of a tertiary care hospital in North India. Materials and Methods: The conjunctival swabs were collected from 23 suspected adenoviral conjunctivitis patients between April-August, 2014 and transported in viral transport media. The samples were subjected to nested PCR targeting hexon gene of human adenovirus. The band size of 956bp was eluted and 8 representative positive samples were subjected to sequencing. The sequences were analyzed by using CLUSTALX2.1 and MEGA 5.1 software. Results: The male: female ratio was found to be 3.6:1. The mean age of presenting patients was 43.95 years (+17.2). Approximately 52.1% (12/23) of patients presented with bilateral involvement while 47.8% (11/23) with unilateral involvement of the eye. Human adenovirus DNA could be detected in 65.2% (15/23) of the patients. The phylogenetic analysis revealed presence of serotype 8 in 7 patients and serotype 4 in one patient. The serotype 8 sequences showed 99-100% identity with Tunisian, Indian and Japanese strains. The adenovirus serotype 4 strains had 100% identity with strains from Tunisia, China and USA. Conclusion: Human adenovirus was found be an important etiological agent for conjunctivitis in our set up. The phylogenetic analysis showed that the predominant circulating strains in our epidemic keratoconjunctivitis were serotypes 8 and 4.Keywords: conjunctivitis, human adenovirus, molecular epidemiology, phylogenetics
Procedia PDF Downloads 283801 Design and Optimisation of 2-Oxoglutarate Dioxygenase Expression in Escherichia coli Strains for Production of Bioethylene from Crude Glycerol
Authors: Idan Chiyanzu, Maruping Mangena
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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization
Procedia PDF Downloads 280800 Acupoint Injection of High Concentration of Glucose Attenuates Mice Chronic Pain and Depression Comorbidity
Authors: Chanya Inprasit, Yi-Wen Lin
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Inflammation causes changes of peripheral and central nervous system properties, affecting both neuronal and non-neuronal cells, resulting in inflammatory pain. Acupoint injection (AI) was developed in the 1950s and has been widely used for relieving pain. It is an acupoint-stimulating technique that utilizes anatomically based meridians derived from Chinese medicine theory. AI has been accepted as an effective treatment and is thought to display superior results when compared to traditional acupuncture methods. However, the mechanism of AI needs to be ratified by more scientific evidence in order to support the theory and its therapeutic development. In this study, we explored the effect of AI on the comorbidity of chronic pain and depression. Mice hindpaw was injected by complete Freund’s adjuvant (CFA) to induce the condition of chronic pain. Measurements of mechanical and thermal hyperalgesia and depression-like behavior were analyzed. The results indicated a positive tendency to AI treatment. The comorbidity of chronic pain and depression was investigated with relation to transient receptor potential V1 (TRPV1) mechanism through the use of TRPV1 gene deletion. The expression of nociceptors such as voltage-gated sodium channels (Navs) or TRPV1, was significantly down-regulated by AI. The expression of inflammation-activated molecules: astrocytic marker glial fibrillary acidic protein (GFAP), the microglial marker Iba-1, S100B, and related kinases, were reversed by AI in both the peripheral and central nervous system. Taken together, these data provided a detailed molecular mechanism of AI-induced analgesia and anti-inflammatory properties. This finding may be utilized for clinical practice to treat chronic pain and depression comorbidity.Keywords: inflammatory pain, acupoint injection, TRPV1, GFAP, S100B
Procedia PDF Downloads 154799 Anti-Aging Effects of Retinol and Alpha Hydroxy Acid on Elastin Fibers of Artificially Photo-Aged Human Dermal Fibroblast Cell Lines
Authors: Mohammed Jarrar, Shalini Behl, Nadia Shaheen, Abeer Fatima, Reem Nasab
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Skin aging is a slow multifactorial process influenced by both internal as well as external factors. Ultra-violet radiations (UV), diet, smoking and personal habits are the most common environmental factors that affect skin aging. Fat contents and fibrous proteins as collagen and elastin are core internal structural components. The direct influence of UV on elastin integrity and health is crucial on aging of skin by time. The deposition of abnormal elastic material is a major marker in a photo-aged skin. Searching for compounds that may protect against cutaneous photo-damage is highly valued. Retinoids and Alpha Hydroxy Acids protective and or repairing effects of UV have been endorsed by some researchers. For consolidating a better understanding of anti and protective effects of such anti-aging agents, we evaluated the combinatory effects of various dosages of lactic acid and retinol on the dermal fibroblasts elastin levels exposed to UV. The UV exposed cells showed significant reduction in the elastin levels. A combination of drugs with a higher concentration of lactic acid (30-35 mM) and a lower concentration of retinol (10-15mg/mL) showed to work better in enhancing elastin concentration in UV exposed cells. We assume this enhancement could be the result of increased tropo-elastin gene expression stimulated by retinol and lactic acid probably repaired the UV irradiated damage by enhancing the amount and integrity of the elastin fibers.Keywords: alpha hydroxy acid, elastin, retinol, ultraviolet radiations
Procedia PDF Downloads 346798 Combining Transcriptomics, Bioinformatics, Biosynthesis Networks and Chromatographic Analyses for Cotton Gossypium hirsutum L. Defense Volatiles Study
Authors: Ronald Villamar-Torres, Michael Staudt, Christopher Viot
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Cotton Gossypium hirsutum L. is one of the most important industrial crops, producing the world leading natural textile fiber, but is very prone to arthropod attacks that reduce crop yield and quality. Cotton cultivation, therefore, makes an outstanding use of chemical pesticides. In reaction to herbivorous arthropods, cotton plants nevertheless show natural defense reactions, in particular through volatile organic compounds (VOCs) emissions. These natural defense mechanisms are nowadays underutilized but have a very high potential for cotton cultivation, and elucidating their genetic bases will help to improve their use. Simulating herbivory attacks by mechanical wounding of cotton plants in greenhouse, we studied by qPCR the changes in gene expression for genes of the terpenoids biosynthesis pathway. Differentially expressed genes corresponded to higher levels of the terpenoids biosynthesis pathway and not to enzymes synthesizing particular terpenoids. The genes were mapped on the G. hirsutum L. reference genome; their global relationships inside the general metabolic pathways and the biosynthesis of secondary metabolites were visualized with iPath2. The chromatographic profiles of VOCs emissions indicated first monoterpenes and sesquiterpenes emissions, dominantly four molecules known to be involved in plant reactions to arthropod attacks. As a result, the study permitted to identify potential key genes for the emission of volatile terpenoids by cotton plants in reaction to an arthropod attack, opening possibilities for molecular-assisted cotton breeding in benefit of smallholder cotton growers.Keywords: biosynthesis pathways, cotton, mechanisms of plant defense, terpenoids, volatile organic compounds
Procedia PDF Downloads 380797 Characterization and Pcr Detection of Selected Strains of Psychrotrophic Bacteria Isolated From Raw Milk
Authors: Kidane workelul, Li xu, Xiaoyang Pang, Jiaping Lv
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Dairy products are exceptionally ideal media for the growth of microorganisms because of their high nutritional content. There are several ways that milk might get contaminated throughout the milking process, including how the raw milk is transported and stored, as well as how long it is kept before being processed. Psychrotrophic bacteria are among the one which can deteriorate the quality of milk mainly their heat resistance proteas and lipase enzyme. For this research purpose 8 selected strains of Psychrotrophic bacteria (Entrococcus hirae, Pseudomonas fluorescens, Pseudomonas azotoformans, Pseudomonas putida, Exiguobacterium indicum, Pseudomonas paralactice, Acinetobacter indicum, Serratia liquefacients)are chosen and try to determine their characteristics based on the research methodology protocol. Thus, the 8 selected strains are cultured, plated incubate, extracted their genomic DNA and genome DNA was amplified, the purpose of the study was to identify their Psychrotrophic properties, lipase hydrolysis positive test, their optimal incubation temperature, designed primer using the noble strain P,flourescens conserved region area in target with lipA gene, optimized primer specificity as well as sensitivity and PCR detection for lipase positive strains using the design primers. Based on the findings both the selected 8 strains isolated from stored raw milk are Psychrotrophic bacteria, 6 of the selected strains except the 2 strains are positive for lipase hydrolysis, their optimal temperature is 20 to 30 OC, the designed primer specificity is very accurate and amplifies for those strains only with lipase positive but could not amplify for the others. Thus, the result is promising and could help in detecting the Psychrotrophic bacteria producing heat resistance enzymes (lipase) at early stage before the milk is processed and this will safe production loss for the dairy industry.Keywords: dairy industry, heat-resistant, lipA, milk, primer and psychrotrophic
Procedia PDF Downloads 68796 Assessment of Mountain Hydrological Processes in the Gumera Catchment, Ethiopia
Authors: Tewele Gebretsadkan Haile
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Mountain terrains are essential to regional water resources by regulating hydrological processes that use downstream water supplies. Nevertheless, limited observed earth data in complex topography poses challenges for water resources regulation. That's why satellite product is implemented in this study. This study evaluates hydrological processes on mountain catchment of Gumera, Ethiopia using HBV-light model with satellite precipitation products (CHIRPS) for the temporal scale of 1996 to 2010 and area coverage of 1289 km2. The catchment is characterized by cultivation dominant and elevation ranges from 1788 to 3606 m above sea level. Three meteorological stations have been used for downscaling of the satellite data and one stream flow for calibration and validation. The result shows total annual water balance showed that precipitation 1410 mm, simulated 828 mm surface runoff compared to 1042 mm observed stream flow with actual evapotranspiration estimate 586mm and 1495mm potential evapotranspiration. The temperature range is 9°C in winter to 21°C. The catchment contributes 74% as quack runoff to the total runoff and 26% as lower groundwater storage, which sustains stream flow during low periods. The model uncertainty was measured using different metrics such as coefficient of determination, model efficiency, efficiency for log(Q) and flow weighted efficiency 0.76, 0.74, 0.66 and 0.70 respectively. The research result highlights that HBV model captures the mountain hydrology simulation and the result indicates quack runoff due to the traditional agricultural system, slope factor of the topography and adaptation measure for water resource management is recommended.Keywords: mountain hydrology, CHIRPS, Gumera, HBV model
Procedia PDF Downloads 22795 Quantification of the Gumera Catchment's Mountain Hydrological Processes in Ethiopia
Authors: Tewele Gebretsadkan Haile
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Mountain terrains are essential to regional water resources by regulating hydrological processes that use downstream water supplies. Nevertheless, limited observed earth data in complex topography poses challenges for water resources regulation. That's why satellite product is implemented in this study. This study evaluates hydrological processes on mountain catchment of Gumera, Ethiopia using HBV-light model with satellite precipitation products (CHIRPS) for the temporal scale of 1996 to 2010 and area coverage of 1289 km2. The catchment is characterized by cultivation dominant and elevation ranges from 1788 to 3606 m above sea level. Three meteorological stations have been used for downscaling of the satellite data and one stream flow for calibration and validation. The result shows total annual water balance showed that precipitation 1410 mm, simulated 828 mm surface runoff compared to 1042 mm observed stream flow with actual evapotranspiration estimate 586mm and 1495mm potential evapotranspiration. The temperature range is 9°C in winter to 21°C. The catchment contributes 74% as quack runoff to the total runoff and 26% as lower groundwater storage, which sustains stream flow during low periods. The model uncertainty was measured using different metrics such as coefficient of determination, model efficiency, efficiency for log(Q) and flow weighted efficiency 0.76, 0.74, 0.66 and 0.70 respectively. The research result highlights that HBV model captures the mountain hydrology simulation and the result indicates quack runoff due to the traditional agricultural system, slope factor of the topography and adaptation measure for water resource management is recommended.Keywords: mountain hydrology, CHIRPS, HBV model, Gumera
Procedia PDF Downloads 16794 A Review of Protocols and Guidelines Addressing the Exposure of Occupants to Electromagnetic Field (EMF) Radiation in Buildings
Authors: Shabnam Monadizadeh, Charles Kibert, Jiaxuan Li, Janghoon Woo, Ashish Asutosh, Samira Roostaei, Maryam Kouhirostami
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A significant share of the technology that has emerged over the past several decades produces electromagnetic field (EMF) radiation. Communications devices, household appliances, industrial equipment, and medical devices all produce EMF radiation with a variety of frequencies, strengths, and ranges. Some EMF radiation, such as Extremely Low Frequency (ELF), Radio Frequency (RF), and the ionizing range have been shown to have harmful effects on human health. Depending on the frequency and strength of the radiation, EMF radiation can have health effects at the cellular level as well as at brain, nervous, and cardiovascular levels. Health authorities have enacted regulations locally and globally to set critical values to limit the adverse effects of EMF radiation. By introducing a more comprehensive field of EMF radiation study and practice, architects and designers can design for a safer electromagnetic (EM) indoor environment, and, as building and construction specialists, will be able to monitor and reduce EM radiation. This paper identifies the nature of EMF radiation in the built environment, the various EMF radiation sources, and its human health effects. It addresses European and US regulations for EMF radiation in buildings and provides a preliminary action plan. The challenges of developing measurement protocols for the various EMF radiation frequency ranges and determining the effects of EMF radiation on building occupants are discussed. This paper argues that a mature method for measuring EMF radiation in building environments and linking these measurements to human health impacts occupant health should be developed to provide adequate safeguards for human occupants of buildings for future research.Keywords: biological affection, electromagnetic field, building regulation, human health, healthy building, clean construction
Procedia PDF Downloads 188793 Spectrum and Prevalence of Candida Infection in Diabetic Foot Ulcers
Authors: Seyed Reza Aghili, Tahereh Shokohi, Lotfollah Davoodi, Zahra Kashi, Azam Moslemi, Mahdi Abastabar, Iman Haghani, Sabah Mayahi, Asoudeh A.
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Introduction: In diabetic foot ulcers, if fungal agents such as Candida species penetrate into the cutaneous or depth of ulcer, can increase the degree of the wound and cause Candia infection and make it more difficult to heal. Material & Methods: A cross-sectional study was performed on 100 diabetic foot ulcer patients in 2020 in Sari, Iran. patient's data and wound grade were recorded in a questionnaire. Candida infection was diagnosed with direct microscopic examination and culture of samples. Colony-PCR molecular method was used for ITS region of DNA and then PCR-RFLP with Msp1 enzyme and using HWP1 specific gene to determine species of Candida agent. Results: Of 100 patients, the mean age 62.1 ± 10.8 years, 95% type 2 diabetes, 83%>10 years duration diabetes, 59% male, 66%> poor education level, 99% married, 52% rural, 95% neuropathic symptoms, 88% using antibiotics, 69%HbA1C >9%, and mean ulcer degree 2.6±1.05 were. Candida infection was seen in 13% of the deep tissue of the wound and 7% cutaneous around the wound. The predominant Candida isolated was C. parapsilosis (71.5%), C .albicans (14.3%). Fungal infections caused by mold fungi were not detected. There was a statistically significant relationship between yeast infection and gender, rural, HbA1C and ulcer degree. Conclusion: Mycological evaluations often are ignored. Candida parapsilosis is the most common infectious agent in these patients and may require specific treatment. Therefore, more attention of physicians to Candida infections particularly, early diagnosis and effective treatment can help faster recovery and prevent amputation.Keywords: diabetic foot ulcer, candida infection, risk factors, c. parapsilosis
Procedia PDF Downloads 204792 Applied Spatial Mapping and Monitoring of Illegal Landfills for Deprived Urban Areas in Romania
Authors: Șercăianu Mihai, Aldea Mihaela, Iacoboaea Cristina, Luca Oana, Nenciu Ioana
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The rise and mitigation of unauthorized illegal waste dumps are a significant global issue within waste management ecosystems, impacting disadvantaged communities. Globally, including in Romania, many individuals live in houses without legal recognition, lacking ownership or construction permits, in areas known as "informal settlements." An increasing number of regions and cities in Romania are struggling to manage their illegal waste dumps, especially in the context of increasing poverty and lack of regulation related to informal settlements. One such informal settlement is located at the end of Bistra Street in Câlnic, within the Reșița Municipality of Caras Severin County. The article presents a case study that focuses on employing remote sensing techniques and spatial data to monitor and map illegal waste practices, with subsequent integration into a geographic information system tailored for the Reșița community. In addition, the paper outlines the steps involved in devising strategies aimed at enhancing waste management practices in disadvantaged areas, aligning with the shift toward a circular economy. Results presented in the paper contain a spatial mapping and visualization methodology calibrated with in situ data collection applicable for identifying illegal landfills. The emergence and neutralization of illegal dumps pose a challenge in the field of waste management. These approaches, which prove effective where conventional solutions have failed, need to be replicated and adopted more wisely.Keywords: informal settlements, GIS, waste dumps, waste management, monitoring
Procedia PDF Downloads 92791 Economic and Financial Crime, Forensic Accounting and Sustainable Developments Goals (SDGs). Bibliometric Analysis
Authors: Monica Violeta Achim, Sorin Nicolae Borlea
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This aim of this work is to stress the needs for enhancing the role of forensic accounting in fighting economic and financial crime, in the context of the new international regulation movements in this area enhanced by the International Federation of Accountants (IFAC). Corruption, money laundering, tax evasion and other frauds significant hamper the economic growth and human development and, ultimately, the UN Sustainable Development Goals. The present paper also stresses the role of good governance in fighting the frauds, in order to achieve the most suitable sustainable development of the society. In this view, we made a bibliometric systematic review on forensic accounting and its contribution towards fraud detection and prevention and theirs relationship with good governance and Sustainable Developments Goals (SDGs). In this view, two powerful bibliometric visual software tools, VosViewer and CiteSpace are used in order to analyze published papers identifies in Scopus and Web of Science databases over the time. Our findings reveal the main red flags identified in literature as used tools by forensic accounting, the evolution in time of the interest of the topic, the distribution in space among world countries and connectivity with patterns of a good governance. Visual designs and scientific maps are useful to show these findings, in a visual way. Our findings are useful for managers and policy makers to provide important avenues that may help in reaching the 2030 Agenda for Sustainable Development, adopted by all United Nations Member States in 2015, in the area of using forensic accounting in preventing frauds.Keywords: forensic accounting, frauds, red flags, SDGs
Procedia PDF Downloads 145790 An Experimental Test of the Effects of Acute and Chronic Stress on Maternal Sensitivity
Authors: Mindy A. Brown, Emma E. Reardon, Jennifer Isenhour, Sheila E. Crowell, K. Lee Raby, Elisabeth Conradt
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The positive impact of maternal sensitivity on infant social and emotional development is well-known, as is the notion that stress may impair a mother’s ability to provide sensitive care for her infant. However, individual differences in susceptibility to parenting-related stress are less understood. This study explores how chronic prenatal stress moderates the effect of acute stressors on maternal sensitivity. Data were gathered from 110 mothers and their 7-month-old infants. Mothers were exposed to either an acute stress task or a control task, after which they engaged in the still-face paradigm, a face-to-face interaction where maternal sensitivity was measured. Chronic maternal stress was assessed using the UCLA Life Stress Interview during the third trimester of pregnancy. The results revealed that among mothers exposed to the stress condition, those with higher chronic stress levels in the previous six months displayed significantly lower sensitivity during the still-face paradigm compared to those with lower chronic stress. Notably, past stress levels had no effect on maternal sensitivity in the control condition. These findings suggest a moderating effect of chronic stress on maternal caregiving behavior, with higher prenatal stress diminishing a mother’s ability to cope with acute parenting-related stressors in the present. The mechanisms behind this may involve changes in stress reactivity pathways, such as the hypothalamic-pituitary-adrenal (HPA) axis or altered emotion regulation strategies developed in response to chronic stress. Understanding these pathways could guide targeted interventions for mothers who may be more vulnerable to stress, improving caregiving outcomes.Keywords: acute stress, maternal stress, prenatal stress, still-face paradigm
Procedia PDF Downloads 33789 Detection and Expression of Peroxidase Genes in Trichoderma harzianum KY488466 and Its Response to Crude Oil Degradation
Authors: Michael Dare Asemoloye, Segun Gbolagade Jonathan, Rafiq Ahmad, Odunayo Joseph Olawuyi, D. O. Adejoye
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Fungi have potentials for degrading hydrocarbons through the secretion of different enzymes. Crude oil tolerance and degradation by Trichoderma harzianum was investigated in this study with its ability to produce peroxidase enzymes (LiP and MnP). Many fungal strains were isolated from rhizosphere of grasses growing on a crude oil spilled site, and the most frequent strain based on percentage incidence was further characterized using morphological and molecular characteristics. Molecular characterization was done through the amplification of Ribosomal-RNA regions of 18s (1609-1627) and 28s (287-266) using ITS1 and ITS4 combinations and it was identified using NCBI BLAST tool. The selected fungus was also subjected to an in-vitro tolerance test at crude oil concentrations of 5, 10, 15, 20 and 25% while 0% served as control. In addition, lignin peroxidase genes (lig1-6) and manganese peroxidase gene (mnp) were detected and expressed in this strain using RT-PCR technique, its peroxidase producing activities was also studied in aliquots (U/ml). This strain had highest incidence of 80%, it was registered in NCBI as Trichoderma harzianum asemoJ KY488466. The strain KY488466 responded to crude oil concentrations as it increase, the dose inhibition response percentage (DIRP) increased from 41.67 to 95.41 at 5 to 25 % crude oil concentrations. All the peroxidase genes are present in KY488466, and expressed with amplified 900-1000 bp through RT-PCR technique. In this strain, lig2, lig4 and mnp genes were over-expressed, lig 6 was moderately expressed, while none of the genes was under-expressed. The strain also produced 90±0.87 U/ml lignin peroxidase and 120±1.23 U/mil manganese peroxidase enzymes in aliquots. These results imply that KY488466 can tolerate and survive high crude oil concentration and could be exploited for bioremediation of oil-spilled soils, the produced peroxidase enzymes could also be exploited for other biotechnological experiments.Keywords: crude oil, enzymes, expression, peroxidase genes, tolerance, Trichoderma harzianum
Procedia PDF Downloads 234788 Digital Athena – Contemporary Commentaries and Greek Mythology Explored through 3D Printing
Authors: Rose Lastovicka, Bernard Guy, Diana Burton
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Greek myth and art acted as tools to think with, and a lens through which to explore complex topics as a form of social media. In particular, coins were a form of propaganda to communicate the wealth and power of the city-states they originated from as they circulated from person to person. From this, how can the application of 3D printing technologies explore the infusion of ancient forms with contemporary commentaries to promote discussion? The digital reconstruction of artifacts is a topic that has been researched by various groups all over the globe. Yet, the exploration of Greek myth through artifacts infused with contemporary issues is currently unexplored in this medium. Using the Stratasys J750 3D printer - a multi-material, full-colour 3D printer - a series of coins inspired by ancient Greek currency and myth was created to present commentaries on the adversities surrounding individuals in the LGBT+ community. Using the J750 as the medium for expression allows for complete control and precision of the models to create complex high-resolution iconography. The coins are printed with a hard, translucent material with coloured 3D visuals embedded into the coin to then be viewed in close contact by the audience. These coins as commentaries present an avenue for wider understanding by drawing perspectives not only from sources concerned with the contemporary LGBT+ community but also from sources exploring ancient homosexuality and the perception and regulation of it in antiquity. By displaying what are usually points of contention between anti- and pro-LGBT+ parties, this visual medium opens up a discussion to both parties, suggesting heritage can play a vital interpretative role in the contemporary world.Keywords: 3D printing, design, Greek mythology, LGBT+ community
Procedia PDF Downloads 122787 Impacts of Hydrologic and Topographic Changes on Water Regime Evolution of Poyang Lake, China
Authors: Feng Huang, Carlos G. Ochoa, Haitao Zhao
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Poyang Lake, the largest freshwater lake in China, is located at the middle-lower reaches of the Yangtze River basin. It has great value in socioeconomic development and is internationally recognized as an important lacustrine and wetland ecosystem with abundant biodiversity. Impacted by ongoing climate change and anthropogenic activities, especially the regulation of the Three Gorges Reservoir since 2003, Poyang Lake has experienced significant water regime evolution, resulting in challenges for the management of water resources and the environment. Quantifying the contribution of hydrologic and topographic changes to water regime alteration is necessary for policymakers to design effective adaption strategies. Long term hydrologic data were collected and the back-propagation neural networks were constructed to simulate the lake water level. The impacts of hydrologic and topographic changes were differentiated through scenario analysis that considered pre-impact and post-impact hydrologic and topographic scenarios. The lake water regime was characterized by hydrologic indicators that describe monthly water level fluctuations, hydrologic features during flood and drought seasons, and frequency and rate of hydrologic variations. The results revealed different contributions of hydrologic and topographic changes to different features of the lake water regime.Noticeable changes were that the water level declined dramatically during the period of reservoir impoundment, and the drought was enhanced during the dry season. The hydrologic and topographic changes exerted a synergistic effect or antagonistic effect on different lake water regime features. The findings provide scientific reference for lacustrine and wetland ecological protection associated with water regime alterations.Keywords: back-propagation neural network, scenario analysis, water regime, Poyang Lake
Procedia PDF Downloads 143786 A Secreted Protein Can Attenuate High Fat Diet Induced Obesity and Metabolic Syndrome in Mice
Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler
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Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome
Procedia PDF Downloads 354785 Biophysical Study of the Interaction of Harmalol with Nucleic Acids of Different Motifs: Spectroscopic and Calorimetric Approaches
Authors: Kakali Bhadra
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Binding of small molecules to DNA and recently to RNA, continues to attract considerable attention for developing effective therapeutic agents for control of gene expression. This work focuses towards understanding interaction of harmalol, a dihydro beta-carboline alkaloid, with different nucleic acid motifs viz. double stranded CT DNA, single stranded A-form poly(A), double-stranded A-form of poly(C)·poly(G) and clover leaf tRNAphe by different spectroscopic, calorimetric and molecular modeling techniques. Results of this study converge to suggest that (i) binding constant varied in the order of CT DNA > poly(C)·poly(G) > tRNAphe > poly(A), (ii) non-cooperative binding of harmalol to poly(C)·poly(G) and poly(A) and cooperative binding with CT DNA and tRNAphe, (iii) significant structural changes of CT DNA, poly(C)·poly(G) and tRNAphe with concomitant induction of optical activity in the bound achiral alkaloid molecules, while with poly(A) no intrinsic CD perturbation was observed, (iv) the binding was predominantly exothermic, enthalpy driven, entropy favoured with CT DNA and poly(C)·poly(G) while it was entropy driven with tRNAphe and poly(A), (v) a hydrophobic contribution and comparatively large role of non-polyelectrolytic forces to Gibbs energy changes with CT DNA, poly(C)·poly(G) and tRNAphe, and (vi) intercalated state of harmalol with CT DNA and poly(C)·poly(G) structure as revealed from molecular docking and supported by the viscometric data. Furthermore, with competition dialysis assay it was shown that harmalol prefers hetero GC sequences. All these findings unequivocally pointed out that harmalol prefers binding with ds CT DNA followed by ds poly(C)·poly(G), clover leaf tRNAphe and least with ss poly(A). The results highlight the importance of structural elements in these natural beta-carboline alkaloids in stabilizing different DNA and RNA of various motifs for developing nucleic acid based better therapeutic agents.Keywords: calorimetry, docking, DNA/RNA-alkaloid interaction, harmalol, spectroscopy
Procedia PDF Downloads 228784 Entry Inhibitors Are Less Effective at Preventing Cell-Associated HIV-2 Infection than HIV-1
Authors: A. R. Diniz, P. Borrego, I. Bártolo, N. Taveira
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Cell-to-cell transmission plays a critical role in the spread of HIV-1 infection in vitro and in vivo. Inhibition of HIV-1 cell-associated infection by antiretroviral drugs and neutralizing antibodies (NAbs) is more difficult compared to cell-free infection. Limited data exists on cell-associated infection by HIV-2 and its inhibition. In this work, we determined the ability of entry inhibitors to inhibit HIV-1 and HIV-2 cell-to cell fusion as a proxy to cell-associated infection. We developed a method in which Hela-CD4-cells are first transfected with a Tat expressing plasmid (pcDNA3.1+/Tat101) and infected with recombinant vaccinia viruses expressing either the HIV-1 (vPE16: from isolate HTLV-IIIB, clone BH8, X4 tropism) or HIV-2 (vSC50: from HIV-2SBL/ISY, R5 and X4 tropism) envelope glycoproteins (M.O.I.=1 PFU/cell).These cells are added to TZM-bl cells. When cell-to-cell fusion (syncytia) occurs the Tat protein diffuses to the TZM-bl cells activating the expression of a reporter gene (luciferase). We tested several entry inhibitors including the fusion inhibitors T1249, T20 and P3, the CCR5 antagonists MVC and TAK-779, the CXCR4 antagonist AMD3100 and several HIV-2 neutralizing antibodies (Nabs). All compounds inhibited HIV-1 and HIV-2 cell fusion albeit to different levels. Maximum percentage of HIV-2 inhibition (MPI) was higher for fusion inhibitors (T1249- 99.8%; P3- 95%, T20-90%) followed by co-receptor antagonists (MVC- 63%; TAK-779- 55%; AMD3100- 45%). NAbs from HIV-2 infected patients did not prevent cell fusion up to the tested concentration of 4μg/ml. As for HIV-1, MPI reached 100% with TAK-779 and T1249. For the other antivirals, MPIs were: P3-79%; T20-75%; AMD3100-61%; MVC-65%.These results are consistent with published data. Maraviroc had the lowest IC50 both for HIV-2 and HIV-1 (IC50 HIV-2= 0.06 μM; HIV-1=0.0076μM). Highest IC50 were observed with T20 for HIV-2 (3.86μM) and with TAK-779 for HIV-1 (12.64μM). Overall, our results show that entry inhibitors in clinical use are less effective at preventing Env mediated cell-to-cell-fusion in HIV-2 than in HIV-1 which suggests that cell-associated HIV-2 infection will be more difficult to inhibit compared to HIV-1. The method described here will be useful to screen for new HIV entry inhibitors.Keywords: cell-to-cell fusion, entry inhibitors, HIV, NAbs, vaccinia virus
Procedia PDF Downloads 312783 Mechanisms and Regulation of the Bi-directional Motility of Mitotic Kinesin Nano-motors
Authors: Larisa Gheber
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Mitosis is an essential process by which duplicated genetic information is transmitted from mother to daughter cells. Incorrect chromosome segregation during mitosis can lead to genetic diseases, chromosome instability and cancer. This process is mediated by a dynamic microtubule-based intracellular structure, the mitotic spindle. One of the major factors that govern the mitotic spindle dynamics are the kinesin-5 biological nano motors that were believed to move unidirectionally on the microtubule filaments, using ATP hydrolysis, thus performing essential functions in mitotic spindle dynamics. Surprisingly, several reports from our and other laboratories have demonstrated that some kinesin-5 motors are bi-directional: they move in minus-end direction on the microtubules as single-molecules and can switch directionality under a number of conditions. These findings broke a twenty-five-years old dogma regarding kinesin directionality (1, 2). The mechanism of this bi-directional motility and its physiological significance remain unclear. To address this unresolved problem, we apply an interdisciplinary approach combining live cell imaging, biophysical single molecule, and structural experiments to examine the activity of these motors and their mutated variants in vivo and in vitro. Our data shows that factors such as protein phosphorylation (3, 4), motor clustering on the microtubules (5, 6) and structural elements (7, 8) regulate the bi-directional motility of kinesin motors. We also show, using Cryo-EM, that bi-directional kinesin motors obtain non-canonical microtubule binding, which is essential to their special motile properties and intracellular functions. We will discuss the implication of these findings to mechanism bi-directional motility and physiological roles in mitosis.Keywords: mitosis, cancer, kinesin, microtubules, biochemistry, biophysics
Procedia PDF Downloads 85782 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells
Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu
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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system
Procedia PDF Downloads 425781 Investigation of Effective Parameters on Pullout Capacity in Soil Nailing with Special Attention to International Design Codes
Authors: R. Ziaie Moayed, M. Mortezaee
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An important and influential factor in design and determining the safety factor in Soil Nailing is the ultimate pullout capacity, or, in other words, bond strength. This important parameter depends on several factors such as material and soil texture, method of implementation, excavation diameter, friction angle between the nail and the soil, grouting pressure, the nail depth (overburden pressure), the angle of drilling and the degree of saturation in soil. Federal Highway Administration (FHWA), a customary regulation in the design of nailing, is considered only the effect of the soil type (or rock) and the method of implementation in determining the bond strength, which results in non-economic design. The other regulations are each of a kind, some of the parameters affecting bond resistance are not taken into account. Therefore, in the present paper, at first the relationships and tables presented by several valid regulations are presented for estimating the ultimate pullout capacity, and then the effect of several important factors affecting on ultimate Pullout capacity are studied. Finally, it was determined, the effect of overburden pressure (in method of injection with pressure), soil dilatation and roughness of the drilling surface on pullout strength is incremental, and effect of degree of soil saturation on pullout strength to a certain degree of saturation is increasing and then decreasing. therefore it is better to get help from nail pullout-strength test results and numerical modeling to evaluate the effect of parameters such as overburden pressure, dilatation, and degree of soil saturation, and so on to reach an optimal and economical design.Keywords: soil nailing, pullout capacity, federal highway administration (FHWA), grout
Procedia PDF Downloads 154780 True Selves: Emerging in Emergency through Clinical Dance Therapy
Authors: Ambra B. Bergamasco
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This paper presents research findings from 35 Clinical Dance Therapy sessions conducted with migrant women aged 24 to 55 from North and Sub-Saharan Africa residing in hosting facilities in Northern Italy. It explores trauma arising from forced displacement, dispossession, and perilous migration journeys undertaken as survival strategies. While the therapeutic discourse on refugee trauma typically centers on post-traumatic stress disorder (PTSD), this study shifts focus to the transformative potential of embodied therapies in trauma recovery. Clinical Dance Therapy (CDT) offers an integrative, non-verbal modality that reconnects individuals to their physicality, fostering the emergence of the authentic self rather than perpetuating cycles of traumatic repetition. By integrating movement, rhythm, and bodily expression, CDT creates a somatic bridge to agency, self-awareness, and emotional regulation, even within emergency or resource-constrained settings. This paper critically examines the role of embodied therapies in addressing complex trauma, emphasizing their capacity to cultivate resilience and psychological reintegration. Additionally, it discusses the secondary effects on practitioners, highlighting the risks of vicarious traumatization, compassion fatigue, and the need for reflective practice in trauma work. The findings contribute to a holistic understanding of trauma management by demonstrating how dance therapy can support sustainable pathways from trauma victimization to empowered survivorship. The paper concludes by advocating for greater inclusion of embodied and creative approaches in trauma-informed care frameworks for displaced and marginalized populations.Keywords: trauma, clinical dance therapy, refugee trauma, embodied therapies, migrant health
Procedia PDF Downloads 8779 Deficiencies in Vitamin A and Iron Supply Potential of Selected Indigenous Complementary Foods of Infants in Uganda
Authors: Richard Kajjura, Joyce Kikafunda, Roger Whitehead
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Introduction: Indigenous complementary recipes for children (6-23 months) are bulky and inextricably linked. The potential contribution of indigenous complementary foods to infant’s vitamin A and iron needs is not well investigated in Uganda. Less is known whether children in Uganda are living with or without adequate supply of vitamin A and iron nutrients. In this study, vitamin A and iron contents were assessed in the complementary foods fed to infants aged 6-11 months in a Peri-urban setting in Kampala District in Central Uganda. Objective: Assessment of vitamin A and iron contents of indigenous complementary foods of children as fed and associated demographic factor. Method: In a cross sectional study design, one hundred and three (153) households with children aged 6-11 months were randomly selected to participate in the assessment. Complementary food samples were collected from the children’s mothers/caretakers at the time of feeding the child. The mothers’ socio-demographic characteristics of age, education, marital status, occupation and sex collected a semi-qualitative questionnaire. The Vitamin A and iron contents in the complementary foods were analyzed using a UV/VIS spectrophotometer for vitamin A and Atomic Absorption spectrophotometer for iron samples. The data was analyzed using Gene-stat software program. Results: The mean vitamin A content was 97.0± 72.5 µg while that of iron was 1.5 ± 0.4 mg per 100g of food sample as fed. The contribution of indigenous complementary foods found was 32% for vitamin A and 15% iron of the recommended dietary allowance. Age of children was found to be significantly associated Vitamin A and Iron supply potential. Conclusion: The contribution of indigenous complementary foods to infant’s vitamin A and iron needs was low. Complementary foods in Uganda are more likely to be deficient in vitamin A and iron content. Nutrient dense dietary supplementation should be intervened in to make possible for Ugandan children attain full growth potential.Keywords: indigenous complementary food, infant, iron, vitamin A
Procedia PDF Downloads 481778 The Role of a Novel DEAD-Box Containing Protein in NLRP3 Inflammasome Activation
Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu
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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.Keywords: inflammasome, inflammation, innate immunity, pyroptosis
Procedia PDF Downloads 287777 Acoustical Comfort in Major Highway in Birnin Kebbi, Kebbi State-Nigeria
Authors: Muhammad Naziru Yahaya, Mustapha Bashir Ayinde
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Noise has been recognized as a major source of pollution in many urban and semi-urban settlements. Noise pollution causes by vehicular movement in urban cities has reaches an alarming proportion due to continuous increases in vehicles and industrialization. This research aim to determine the geo-physical characteristics of the study area and to determine the level of noise generation and volume intensity in areas where noise levels are high within the metropolis and compare with NESREA and WHO standards. This study identified the various sources of noise, compared noise levels in various parts of the study area with recommended standards and determined the geo-physical characteristic of noise generated. A sound level meter Gm 1352, was used for the noise measurements. The study showed that the noise pollution levels measured in minimum noise level of 63.75 dBA and average maximum of 95.175 dBA, at some locations in Birnin Kebbi metropolis the noise level have exceeded the standard limits set by the World Health Organization (WHO), Federal Environment Protection Agency (FEPA). Results revealed that there was a considerable increase in noise pollution in First Bank roundabout and Haliru Abdu roundabout, attribute to high numbers of vehicular movement and road congestion within Birnin Kebbi. The study therefore concluded that there should be an enforcement and adherence to the regulation regarding noise pollution limit. The minimum average day noise level recorded was 67.225 dBA, and average maximum of 96.6 dBA is an indication that the noise level of Birnin Kebbi metropolis was highly unsatisfactory. Based on this, it is suggested that taking adequate measures and following the laid-down recommendations will reduce traffic noise to the barest minimum.Keywords: decibel, noise level, pollution, sound level, traffic, highway
Procedia PDF Downloads 80776 An Unbiased Profiling of Immune Repertoire via Sequencing and Analyzing T-Cell Receptor Genes
Authors: Yi-Lin Chen, Sheng-Jou Hung, Tsunglin Liu
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Adaptive immune system recognizes a wide range of antigens via expressing a large number of structurally distinct T cell and B cell receptor genes. The distinct receptor genes arise from complex rearrangements called V(D)J recombination, and constitute the immune repertoire. A common method of profiling immune repertoire is via amplifying recombined receptor genes using multiple primers and high-throughput sequencing. This multiplex-PCR approach is efficient; however, the resulting repertoire can be distorted because of primer bias. To eliminate primer bias, 5’ RACE is an alternative amplification approach. However, the application of RACE approach is limited by its low efficiency (i.e., the majority of data are non-regular receptor sequences, e.g., containing intronic segments) and lack of the convenient tool for analysis. We propose a computational tool that can correctly identify non-regular receptor sequences in RACE data via aligning receptor sequences against the whole gene instead of only the exon regions as done in all other tools. Using our tool, the remaining regular data allow for an accurate profiling of immune repertoire. In addition, a RACE approach is improved to yield a higher fraction of regular T-cell receptor sequences. Finally, we quantify the degree of primer bias of a multiplex-PCR approach via comparing it to the RACE approach. The results reveal significant differences in frequency of VJ combination by the two approaches. Together, we provide a new experimental and computation pipeline for an unbiased profiling of immune repertoire. As immune repertoire profiling has many applications, e.g., tracing bacterial and viral infection, detection of T cell lymphoma and minimal residual disease, monitoring cancer immunotherapy, etc., our work should benefit scientists who are interested in the applications.Keywords: immune repertoire, T-cell receptor, 5' RACE, high-throughput sequencing, sequence alignment
Procedia PDF Downloads 198775 Analysis of Extracellular Vesicles Interactomes of two Isoforms of Tau Protein via SHSY-5Y Cell Lines
Authors: Mohammad Aladwan
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Alzheimer’s disease (AD) is a widespread dementing illness with a complex and poorly understood etiology. An important role in improving our understanding of the AD process is the modeling of disease-associated changes in tau protein phosphorylation, a protein known to mediate events essential to the onset and progression of AD. A main feature of AD is the abnormal phosphorylation of tau protein and the presence of neurofibrillary tangles. In order to evaluate the respective roles of the microtubule-binding region (MTBR) and alternatively spliced exons in the N-terminal projection domains in AD, we have constructed SHSY-5Y cell lines that stably overexpress four different species of tau protein (4R2N, 4R0N, N(E-2), N(E+2)). Since the toxicity and spreading of tau lesions in AD depends on the interactions of tau with other proteins, we have performed a proteomic analysis of exosome-fraction interactomes for cell lysates and media samples that were isolated from SHSY-5Y cell lines. Functional analysis of tau interactomes based on gene ontology (GO) terms was performed using the String 10.5 database program. The highest number of exosomes proteomes and tau associated proteins were found with 4R2N isoform (2771 and 159) in cell lysate and they have a high strength of connectivity (78%) between proteins, while N(E-2) isoform in the media proteomes has the highest number of proteins and tau associated protein (1829 and 205). Moreover, known AD markers were significantly enriched in secreted interactomes relative to lysate interactomes in the SHSY-5Y cells of tau isoforms lacking exons 2 and 3 in the N-terminal. The lack of exon 2 (E-2) from tau protein can be mediated by tau secretion and spreading to different cells. Enriched functions in the secreted E-2 interactome include signaling and developmental pathways that have been linked to a) tau misprocessing and lesion development and b) tau secretion and which, therefore, could play novel roles in AD pathogenesis.Keywords: Alzheimer's disease, dementia, tau protein, neurodegenration disease
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