Search results for: oxidative modification of proteins (OMP)
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2574

Search results for: oxidative modification of proteins (OMP)

2244 In Vitro Antioxidant and Free Radical Scavenging Activity of Phyllanthus Emblica L. Extract

Authors: Benyapa Suksuwan

Abstract:

Introduction: Oxidative stress is identified as the root cause of the development and progression of several diseases as the disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Aim of the Study: This study focused on the antioxidant activity of polyphenols extracted from Phyllanthus Emblica L. as oxidative stress plays a vital role in developing and progressing many diseases, including cardiovascular diseases and cancer. Materials and Methods: The plant was extracted using a mixture solvent (ethyl alcohol: water in ratio 8:2). The total phenolic content of P. Emblica extract was determined using the Folin-Cioucalteu method and calculated as gallic acid equivalents (GAE) and various antioxidant assays DPPH and ABTS radical scavenging capacity assays. Results and Discussion: The findings exhibited a strong correlation between antioxidant activity and the total phenol contents. In addition, the IC₅₀ of P. Emblica extract via DPPH and ABTS assays were 68.10 μg/mL ± 0.455, and 49.24 μg/mL ± 0.716, respectively. Furthermore, P. Emblica extract showed antioxidant activities in a concentration-dependent manner. Vitamin C was used as a positive control in the DPPH assay, while Trolox was used as a positive control in the ABTS assay. Conclusions: In conclusion, P. Emblica extract consisted of a high amount of total phenolic content, which possesses potent antioxidant activity. However, further antioxidant activity assays using human cell lines such as SOD, ROS, and RNS scavenging assays and in vitro antioxidant experiments should be performed in order.

Keywords: antioxidant, ABTS scavenging, DPPH scavenging assay, total phenol contents assay, Phyllanthus Emblica L

Procedia PDF Downloads 178
2243 Modification of ZnMgO NPs for Improving Device Performance of Quantum Dot Light-emitting Diodes

Authors: Juyon Lee, Myoungjin Park, Jonghoon Kim, Jaekook Ha, Chanhee Lee

Abstract:

We demonstrated a new positive aging methods of QLEDs devices that can apply in large size inkjet printing display. Conventional positive aging method using photo-curable resin remains unclear mechanism of the phenomenon and also there are many limitations to apply large size panels in commercial process. Through the photo acid generator (PAG) in ETL Ink, we achieved 90% of the efficiency of the conventional method and up to 1000h life time stability (T80). This techniques could be applied to next generation of QLEDs panels and also can prove the working mechanism of positive aging in QLED related to modification of ZnMgO NPs.

Keywords: quantum dots, QLED, printing, positive aging, ZnMgO NPs

Procedia PDF Downloads 124
2242 Chemically Modified Chitosan Derivatives with Ameliorated Properties Appropriate for Drug Delivery

Authors: Georgia M. Michailidou, Nina-Maria S. Ainali, Eleftheria C. Xanthopoulou, Dimitrios N. Bikiaris

Abstract:

Polysaccharides are polymeric materials derived from nature. They are extensively used in pharmaceutical technology due to their low cost, their ready availability and their low toxicity. Chitosan is the product derived from the deacetylation of chitin usually obtained from arthropods. It is a linear polysaccharide which is composed of repeated units of N-deacetylated amino groups and some N-acetylated groups residues. Due to its excellent biological properties, it is an attractive natural polymer. It is biocompatible with low toxicity and complete biodegradability. Although it has excellent properties, the chemical modification of its structure results in new derivatives with ameliorated and more improved properties compared to the initial polymer. This is the exact purpose of the present study in which chitosan was modified with three different monomers, namely trans-aconitic acid, succinic anhydride and 2-hydroxyethyl acrylate. In chitosan’s modification with trans aconitic acid, EDC was utilized as an activator of the carboxylic groups of the monomer, and then a coupling reaction with the amino groups took place. Succinic anhydride reacted with chitosan through a ring opening reaction while 2-hydroxyethyl acrylate reacted through the addition of chitosan’s amino group to the double bond of the monomer. Through FTIR and NMR measurements the success of each reaction was confirmed, and the new structures of the derivatives were verified. X-ray diffraction was utilized in order to examine the effect of the modifications in chitosan’s crystallinity. Finally, swelling tests were conducted in order to assess the improved ability of the new polymeric materials to absorb water. Our results support the successful modification of chitosan’s macromolecular chains in all three reactions. Furthermore, the new derivatives appear to be amorphous concerning their crystallinity and have great ability in absorbing water.

Keywords: chitosan, derivatives, modification, polysaccharide

Procedia PDF Downloads 88
2241 Proteomics Associated with Colonization of Human Enteric Pathogen on Solanum lycopersicum

Authors: Neha Bhadauria, Indu Gaur, Shilpi Shilpi, Susmita Goswami, Prabir K. Paul

Abstract:

The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

Procedia PDF Downloads 255
2240 Rainwater Harvesting and Management of Ground Water (Case Study Weather Modification Project in Iran)

Authors: Samaneh Poormohammadi, Farid Golkar, Vahideh Khatibi Sarabi

Abstract:

Climate change and consecutive droughts have increased the importance of using rainwater harvesting methods. One of the methods of rainwater harvesting and, in other words, the management of atmospheric water resources is the use of weather modification technologies. Weather modification (also known as weather control) is the act of intentionally manipulating or altering the weather. The most common form of weather modification is cloud seeding, which increases rain or snow, usually for the purpose of increasing the local water supply. Cloud seeding operations in Iran have been married since 1999 in central Iran with the aim of harvesting rainwater and reducing the effects of drought. In this research, we analyze the results of cloud seeding operations in the Simindashtplain in northern Iran. Rainwater harvesting with the help of cloud seeding technology has been evaluated through its effects on surface water and underground water. For this purpose, two different methods have been used to estimate runoff. The first method is the US Soil Conservation Service (SCS) curve number method. Another method, known as the reasoning method, has also been used. In order to determine the infiltration rate of underground water, the balance reports of the comprehensive water plan of the country have been used. In this regard, the study areas located in the target area of each province have been extracted by drawing maps of the influence coefficients of each area in the GIS software. It should be mentioned that the infiltration coefficients were taken from the balance sheet reports of the country's comprehensive water plan. Then, based on the area of each study area, the weighted average of the infiltration coefficient of the study areas located in the target area of each province is considered as the infiltration coefficient of that province. Results show that the amount of water extracted from the rain with the help of cloud seeding projects in Simindasht is as follows: an increase in runoff 63.9 million cubic meters (with SCS equation) or 51.2 million cubic meters (with logical equation) and an increase in ground water resources: 40.5 million cubic meters.

Keywords: rainwater harvesting, ground water, atmospheric water resources, weather modification, cloud seeding

Procedia PDF Downloads 85
2239 Quantifying the Protein-Protein Interaction between the Ion-Channel-Forming Colicin A and the Tol Proteins by Potassium Efflux in E. coli Cells

Authors: Fadilah Aleanizy

Abstract:

Colicins are a family of bacterial toxins that kill Escherichia coli and other closely related species. The mode of action of colicins involves binding to an outer membrane receptor and translocation across the cell envelope, leading to cytotoxicity through specific targets. The mechanism of colicin cytotoxicity includes a non-specific endonuclease activity or depolarization of the cytoplasmic membrane by pore-forming activity. For Group A colicins, translocation requires an interaction between the N-terminal domain of the colicin and a series of membrane- bound and periplasmic proteins known as the Tol system (TolB, TolR, TolA, TolQ, and Pal and the active domain must be translocated through the outer membranes. Protein-protein interactions are intrinsic to virtually every cellular process. The transient protein-protein interactions of the colicin include the interaction with much more complicated assemblies during colicin translocation across the cellular membrane to its target. The potassium release assay detects variation in the K+ content of bacterial cells (K+in). This assays is used to measure the effect of pore-forming colicins such as ColA on an indicator organism by measuring the changes of the K+ concentration in the external medium (K+out ) that are caused by cell killing with a K+ selective electrode. One of the goals of this work is to employ a quantifiable in-vivo method to spot which Tol protein are more implicated in the interaction with colicin A as it is translocated to its target.

Keywords: K+ efflux, Colicin A, Tol-proteins, E. coli

Procedia PDF Downloads 384
2238 From Forked Tongues to Tinkerbell Ears: Rethinking the Criminalization of Alternative Body Modification in the UK

Authors: Luci V. Hyett

Abstract:

The criminal law of England and Wales currently deems that a person cannot consent to the infliction of injury upon their own body, where the level of harm is considered to be Actual or Grevious. This renders the defence of consent of the victim as being unavailable to those persons carrying out an Alternative Body Modification procedure. However, the criminalization of consensual injury is more appropriately deemed as being categorized as an offense against public morality and not one against the person, which renders the State’s involvement in the autonomous choices of a consenting adult, when determining what can be done to one’s own body, an arbitrary one. Furthermore, to recognise in law that a person is capable of giving a valid consent to socially acceptable cosmetic interventions that largely consist of procedures designed to aesthetically please men and, not those of people who want to modify their bodies for other reasons means that patriarchal attitudes are continuing to underpin public repulsion and inhibit social acceptance of such practices. Theoretical analysis will begin with a juridical examination of R v M(B) [2019] QB 1 where the High Court determined that Alternative Body Modification was not a special category exempting a person so performing from liability for Grevious Bodily Harm using the defence of consent. It will draw from its reasoning which considered that ‘the removal of body parts were medical procedures being carried out for no medical reason by someone not qualified to carry them out’ which will form the basis of this enquiry. It will consider the philosophical work of Georgio Agamben when analysing whether the biopolitical climate in the UK, which places the optimization of the perfect, healthy body at the centre of political concern can explain why those persons who wish to engage in Alternative Body Modification are treated as the ‘Exception’ to that which is normal using the ‘no medical reason’ canon to justify criminalisation, rather than legitimising the industry through regulation. It will consider, through a feminist lens, the current conflict in law between traditional cosmetic interventions which alter one’s physical appearance for socially accepted aesthetic purposes such as those to the breast, lip and buttock and, modifications described as more outlandish such as earlobe stretching, tooth filing and transdermal implants to create horns and spikes under the skin. It will assert that ethical principles relating to the psychological impact of body modification described as ‘alternative’ is used as a means to exclude person’s seeking such a procedure from receiving safe and competent treatment via a registered cosmetic surgeon which leads to these increasingly popular surgery’s being performed in Tattoo parlours throughout the UK as an extension to other socially acceptable forms of self-modification such as piercings. It will contend that only by ‘inclusive exclusion’ will those ‘othered’ through ostracisation be welcomed into the fold of normality and this can only be achieved through recognition of alternative body modification as a legitimate cosmetic intervention, subject to the same regulatory framework as existing practice. This would assist in refocusing the political landscape by erring on the side of liberty rather than that of biology.

Keywords: biopolitics, body modification, consent, criminal law

Procedia PDF Downloads 90
2237 The Ameliorative Effects of Nanoencapsulated Triterpenoids from Petri-Dish Cultured Antrodia cinnamomea on Reproductive Function of Diabetic Male Rats

Authors: Sabri Sudirman, Yuan-Hua Hsu, Zwe-Ling Kong

Abstract:

Male reproductive dysfunction is predominantly due to insulin resistance and hyperglycemia result in inflammation and oxidative stress. Furthermore, nanotechnology provides an alternative approach to improve the bioavailability of natural active food ingredients. Therefore, the aim of this study were to investigate nanoencapsulated triterpenoids from petri-dish cultured Antrodia cinnamomea (PAC) nanoparticles whether it could increase the bioavailability; in addition, the anti-inflammatory and anti-oxidative effects could more effectively ameliorate the reproductive function of diabetic male rats. First, PAC encapsulated in chitosan-silica nanoparticles (Nano-PAC) were prepared by biosilicification method. Scanning electron micrographs confirm the average particle size is about 30 nm, and the encapsulation efficiency is 83.7% by HPLC. Diabetic male Sprague-Dawley rats were induced by high fat diet (40% kcal from fat) and streptozotocin (35 mg/kg). Nano-PAC was administered by oral gavage in three doses (4, 8 and 20 mg/kg) for 6 weeks. Besides, metformin (300 mg/kg) and nanoparticles (Nano) were treated as the positive and negative control respectively. Results indicated that 4 mg/kg Nano-PAC administration for 6 weeks improved hyperglycemia, insulin resistance, and also reduced advanced glycation end products in plasma. In addition, 8 mg/kg Nano-PAC ameliorated morphological of testicular seminiferous tubules, sperm morphology and motility, reactive oxygen species production and mitochondrial membrane potential. Moreover, 20 mg/kg Nano-PAC restored reproductive endocrine system function and increased KiSS-1 level in plasma. In plasma or testis anti-oxidant superoxide dismutase, glutathione peroxidase and catalase were increased whereas malondialdehyde, as well as pro-inflammatory cytokines tumor necrosis factor-α, interleukin-6, and interferon-gamma, decreased. Most importantly, 8 mg/kg Nano-PAC down-regulated the oxidative stress induced c-Jun N-terminal kinase (JNK) signaling pathway. Our study successfully nanoencapsulated PAC to form nanoparticles and low-dose Nano-PAC improved diabetes-induced hyperglycemia, inflammation and oxidative stress to ameliorate the reproductive function of diabetic male rats.

Keywords: Antrodia cinnamomea, diabetes mellitus, male reproduction, nanoparticles

Procedia PDF Downloads 205
2236 Gallbladder Amyloidosis Causing Gangrenous Cholecystitis: A Case Report

Authors: Christopher Leung, Guillermo Becerril-Martinez

Abstract:

Amyloidosis is a rare systemic disease where abnormal proteins invade various organs and impede their function. Occasionally, they can manifest in a solidary organ such as the heart, lung, and nervous systems; rarely do they manifest in the gallbladder. Diagnosis often requires biopsy of the affected area and histopathology shows deposition of abnormally folded globular proteins called amyloid proteins. This case presents a 69-year-old male with a 3-month history of RUQ pain, diarrhea and non-specific symptoms of tiredness, etc. On imaging, both his US and CT abdomen showed gallbladder wall thickening and pericholecystic fluid, which may represent acute cholecystitis with hypodense lesions around the gallbladder, possibly representing liver abscesses. Given his symptoms of abdominal pain and imaging findings, this gentleman eventually had a laparoscopic cholecystectomy showing a gangrenous gallbladder with a mass on the liver bed. On histopathology, it showed amorphous hyaline eosinophilic material, which Congo-stained confirmed amyloidosis. Amyloidosis explained his non-specific symptoms, he avoided further biopsy, and he was commenced immediately on Lenalidomide. Involvement of the gallbladder is extremely rare, with less than 30 cases around the world. Half of the cases are reported as primary amyloidosis. This case adds to the current literature regarding primary gallbladder amyloidosis. Importantly, this case highlights how laparoscopic cholecystectomy can help with the diagnosis of gallbladder amyloidosis.

Keywords: amyloidosis, cholecystitis, gangrenous cholecystitis, gallbladder, systemic amyloidosis

Procedia PDF Downloads 176
2235 Effect of Modification and Expansion on Emergence of Cooperation in Demographic Multi-Level Donor-Recipient Game

Authors: Tsuneyuki Namekata, Yoko Namekata

Abstract:

It is known that the mean investment evolves from a very low initial value to some high level in the Continuous Prisoner's Dilemma. We examine how the cooperation level evolves from a low initial level to a high level in our Demographic Multi-level Donor-Recipient situation. In the Multi-level Donor-Recipient game, one player is selected as a Donor and the other as a Recipient randomly. The Donor has multiple cooperative moves and one defective move. A cooperative move means the Donor pays some cost for the Recipient to receive some benefit. The more cooperative move the Donor takes, the higher cost the Donor pays and the higher benefit the Recipient receives. The defective move has no effect on them. Two consecutive Multi-level Donor-Recipient games, one as a Donor and the other as a Recipient, can be viewed as a discrete version of the Continuous Prisoner's Dilemma. In the Demographic Multi-level Donor-Recipient game, players are initially distributed spatially. In each period, players play multiple Multi-level Donor-Recipient games against other players. He leaves offspring if possible and dies because of negative accumulated payoff of him or his lifespan. Cooperative moves are necessary for the survival of the whole population. There is only a low level of cooperative move besides the defective move initially available in strategies of players. A player may modify and expand his strategy by his recent experiences or practices. We distinguish several types of a player about modification and expansion. We show, by Agent-Based Simulation, that introducing only the modification increases the emergence rate of cooperation and introducing both the modification and the expansion further increases it and a high level of cooperation does emerge in our Demographic Multi-level Donor-Recipient Game.

Keywords: agent-based simulation, donor-recipient game, emergence of cooperation, spatial structure, TFT, TF2T

Procedia PDF Downloads 347
2234 Antimicrobial Activity of Functionalized Alpaca Fabrics with Silver Nanoparticles

Authors: Gina Zavaleta-Espejo, Segundo R. Jáuregui-Rosas, Fanny V. Samanamud-Moreno, José Saldaña Jiménez, Anibal Felix-Quintero, Víctor Montero-Del Aguila, Elsi Mejía-Uriarte

Abstract:

Vicugnapacos "alpaca" fabrics are considered special for their finesse, and the garments in the textile market are very luxurious. It has many special characteristics such as antiallergic, soft, hygroscopic, among others. In this sense, the research aimed to evaluate the antimicrobial activity of alpaca fabrics functionalized with silver nanoparticles on the bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. For the functionalization of the fabrics, AgNO3 and different concentrations of trisodium citrate (TSC) 2, 6, and 10 mg. Tissue characterization was performed using Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The determination of the antimicrobial activity of the alpaca tissues was made by the Kirby-Bauer method with alpaca tissue discs functionalized with silver nanoparticles, an experimental design was made in completely randomized blocks with three treatments and a negative control with three repetitions. The results showed that inhibition halos were formed for both bacteria, therefore, the functionalized tissues have a high antimicrobial activity, whose mechanism of action is attributed to the free radicals (ROS) generated by the nanoparticles that cause oxidative damage to the bacteria. proteins and lipids of the bacterial cell wall.

Keywords: antimicrobial, animal fibers, fabrics, functionalization, trisodium citrate

Procedia PDF Downloads 113
2233 Aerodynamics and Aeroelastics Studies of Hanger Bridge with H-Beam Profile Using Wind Tunnel

Authors: Matza Gusto Andika, Malinda Sabrina, Syarie Fatunnisa

Abstract:

Aerodynamic and aeroelastics studies on the hanger bridge profile are important to analyze the aerodynamic phenomenon and Aeroelastics stability of hanger. Wind tunnel tests were conducted on a model of H-beam profile from hanger bridge. The purpose of this study is to investigate steady aerodynamic characteristics such as lift coefficient (Cl), drag coefficient (Cd), and moment coefficient (Cm) under the different angle of attack for preliminary prediction of aeroelastics stability problems. After investigation the steady aerodynamics characteristics from the model, dynamic testing is also conducted in wind tunnel to know the aeroelastics phenomenon which occurs at the H-beam hanger bridge profile. The studies show that the torsional vortex induced vibration occur when the wind speed is 7.32 m/s until 9.19 m/s with maximum amplitude occur when the wind speed is 8.41 m/s. The result of wind tunnel testing is matching to hanger vibration where occur in the field, so wind tunnel studies has successful to model the problem. In order that the H-beam profile is not good enough for the hanger bridge and need to be modified to minimize the Aeroelastics problem. The modification can be done with structure dynamics modification or aerodynamics modification.

Keywords: aerodynamics, aeroelastic, hanger bridge, h-beam profile, vortex induced vibration, wind tunnel

Procedia PDF Downloads 327
2232 Protein-Thiocyanate Composite as a Sensor for Iron III Cations

Authors: Hosam El-Sayed, Amira Abou El-Kheir, Salwa Mowafi, Marwa Abou Taleb

Abstract:

Two proteinic biopolymers; namely keratin and sericin, were extracted from their respective natural resources by simple appropriate methods. The said proteins were dissolved in the appropriate solvents followed by regeneration in a form of film polyvinyl alcohol. Proteinium thiocyanate (PTC) composite was prepared by reaction of a regenerated film with potassium thiocyanate in acid medium. In another experiment, the said acidified proteins were reacted with potassium thiocyante before dissolution and regeneration in a form of PTC composite. The possibility of using PTC composite for determination of the concentration of iron III ions in domestic as well as industrial water was examined. The concentration of iron III cations in water was determined spectrophotometrically by measuring the intensity of blood red colour of iron III thiocyanate obtained by interaction of PTC with iron III cation in the tested water sample.

Keywords: iron III cations, protein, sensor, thiocyanate, water

Procedia PDF Downloads 405
2231 The Relationship between Human Neutrophil Elastase Levels and Acute Respiratory Distress Syndrome in Patients with Thoracic Trauma

Authors: Wahyu Purnama Putra, Artono Isharanto

Abstract:

Thoracic trauma is trauma that hits the thoracic wall or intrathoracic organs, either due to blunt trauma or sharp trauma. Thoracic trauma often causes impaired ventilation-perfusion due to damage to the lung parenchyma. This results in impaired tissue oxygenation, which is one of the causes of acute respiratory distress syndrome (ARDS). These changes are caused by the release of pro-inflammatory mediators, plasmatic proteins, and proteases into the alveolar space associated with ongoing edema, as well as oxidative products that ultimately result in severe inhibition of the surfactant system. This study aims to predict the incidence of acute respiratory distress syndrome (ARDS) through human neutrophil elastase levels. This study examines the relationship between plasma elastase levels as a predictor of the incidence of ARDS in thoracic trauma patients in Malang. This study is an observational cohort study. Data analysis uses the Pearson correlation test and ROC curve (receiver operating characteristic curve). It can be concluded that there is a significant (p= 0.000, r= -0.988) relationship between elastase levels and BGA-3. If the value of elastase levels is limited to 23.79 ± 3.95, the patient will experience mild ARDS. While if the value of elastase levels is limited to 57.68 ± 18.55, in the future, the patient will experience moderate ARDS. Meanwhile, if the elastase level is between 107.85 ± 5.04, the patient will likely experience severe ARDS. Neutrophil elastase levels correlate with the degree of severity of ARDS incidence.

Keywords: ARDS, human neutrophil elastase, severity, thoracic trauma

Procedia PDF Downloads 118
2230 Production of Recombinant VP2 Protein of Canine Parvovirus Type 2c Using Baculovirus Expression System

Authors: Jae Young Song, In-Ohk Ouh, Seyeon Park, Byeong Sul Kang, Soo Dong Cho, In-Soo Cho

Abstract:

Canine parvovirus (CPV) is a major pathogen of diarrhea disease in dogs. CPV type 2 has three of antigenic variants such as 2a, 2b, and 2c. CPV constructs a small non-enveloped, icosahedral capsid that contains single-stranded DNA. It has capsids that two largely overlapping virion proteins (VP), VP1 (82 kDa), and VP2 (65 kDa). Baculoviruses are insect pathogens that regulate insect populations in nature and are being successfully used to control insect pests. The proteins produced in the baculovirus-expression system are used for instance for functional studies, vaccine preparations, or diagnostics. The vaccines produced by baculovirus-expression system showed elicitation of antibodies. The recombinant baculovirus infected SF9 cells showed broken shape. The recombinant VP2 proteins from cell pellet or supernatant were confirmed by western blotting. The result showed that the recombinant VP2 protein bands were appeared at 65 kDa molecular weight in both cell pellet and supernatant of infected SF9 cell. These results indicated that the recombinant baculovirus infected SF9 cell express the recombinant VP2 protein successfully. In addition, the expressed recombinant VP2 protein is secreted from cell to supernatant. The baculovirus expression system can be used to produce the VP2 protein of CPV 2c. In addition, the secretion property of the expression of VP2 protein may decrease the cost of production, because it can be skipped the cell breaking step. The produced VP2 protein could be used for vaccine and the agent of diagnostic tests. This study provides the foundation of the production of CPV 2c vaccine and the diagnostic agent.

Keywords: baculovirus, canine parvovirus 2c, dog, Korea

Procedia PDF Downloads 122
2229 Functional Significance of Qatari Camels Milk: Antioxidant Content and Antimicrobial Activity of Protein Fractions

Authors: Tahra ElObeid, Omnya Ahmed, Reem Al-Sharshani, Doaa Dalloul, Jannat Alnattei

Abstract:

Background: Camelus dormedarius camels are also called ‘the Arabian camels’ and are present in the desert area of North Africa and the Middle East. Recently, camel’s milk has a great attention globally because of their proteins and peptides that have been reported to be beneficial for the health and in the management of many diseases. Objectives: This study was designed to investigate the antioxidant, antimicrobial activity and to evaluate the total phenolic content of camel’s milk proteins in Qatar. Methods: Fresh two camel’s milk samples from Omani breed and called Muhajer (camel’s milk A and B) were collected on the 1st of the December. Both samples were from the same location Al- Shahaniyah, Doha, Qatar, but from different local private farms and feeding system. Camel’s milk A and B were defatted by centrifugation and their proteins were extracted by acid and thermal precipitation. The antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. Total phenolic compound (TPC) was evaluated by Folin-Ciocalteu reagent (FCR). On the other hand, the antimicrobial activity against eight different type of pathogenic bacteria was evaluated by disc diffusion method and the zone of inhibition was measured. Results: The of the total phenolic content of whole milk in both camel’s milk A and B were significantly the highest among the protein extracts. The % of the DPPH radical inhibition of casein protein in both camel’s milk A and B were significantly the highest among the protein extracts. In this study, there were marked changes in the antibacterial activity in the different camel milk protein extracts. All extracts showed bacterial overgrowth. Conclusion: The antioxidant activity of the camel milk protein extracts correlated to their unique phenolic compounds and bioactive protein peptides. The antimicrobial activity was not detected perhaps due to the technique, the quality, or the extraction method. Overall, camel's milk exhibits a high antioxidant activity, which is responsible for many health benefits besides the nutritional values.

Keywords: camels milk, antioxidant content, antimicrobial activity, proteins, Qatar

Procedia PDF Downloads 188
2228 CAD Tool for Parametric Design modification of Yacht Hull Surface Models

Authors: Shahroz Khan, Erkan Gunpinar, Kemal Mart

Abstract:

Recently parametric design techniques became a vital concept in the field of Computer Aided Design (CAD), which helps to provide sophisticated platform to the designer in order to automate the design process in efficient time. In these techniques, design process starts by parameterizing the important features of design models (typically the key dimensions), with the implementation of design constraints. The design constraints help to retain the overall shape of the model while modifying its parameters. However, the process of initializing an appropriate number of design parameters and constraints is the crucial part of parametric design techniques, especially for complex surface models such as yacht hull. This paper introduces a method to create complex surface models in favor of parametric design techniques, a method to define the right number of parameters and respective design constraints, and a system to implement design parameters in contract to design constraints schema. For this, in our proposed approach the design process starts by dividing the yacht hull into three sections. Each section consists of different shape lines, which form the overall shape of yacht hull. The shape lines are created using Cubic Bezier Curves, which allow larger design flexibility. Design parameters and constraints are defined on the shape lines in 3D design space to facilitate the designers for better and individual handling of parameters. Afterwards, shape modifiers are developed, which allow the modification of each parameter while satisfying the respective set of criteria and design constraints. Such as, geometric continuities should be maintained between the shape lines of the three sections, fairness of the hull surfaces should be preserved after modification and while design modification, effect of a single parameter should be negligible on other parameters. The constraints are defined individually on shape lines of each section and mutually between the shape lines of two connecting sections. In order to validate and visualize design results of our shape modifiers, a real time graphic interface is created.

Keywords: design parameter, design constraints, shape modifies, yacht hull

Procedia PDF Downloads 280
2227 Neurotoxic Effects Assessment of Metformin in Danio rerio

Authors: Gustavo Axel Elizalde-Velázquez

Abstract:

Metformin is the first line of oral therapy to treat type II diabetes and is also employed as a treatment for other indications, such as polycystic ovary syndrome, cancer, and COVID-19. Recent data suggest it is the aspirin of the 21st century due to its antioxidant and anti-aging effects. However, increasingly current articles indicate its long-term consumption generates mitochondrial impairment. Up to date, it is known metformin increases the biogenesis of Alzheimer's amyloid peptides via up-regulating BACE1 transcription, but further information related to brain damage after its consumption is missing. Bearing in mind the above, this work aimed to establish whether or not chronic exposure to metformin may alter swimming behavior and induce neurotoxicity in Danio rerio adults. For this purpose, 250 Danio rerio grown-ups were assigned to six tanks of 50 L of capacity. Four of the six systems contained 50 fish, while the remaining two had 25 fish (≈1 male:1 female ratio). Every system with 50 fish was allocated one of the three metformin treatment concentrations (1, 20, and 40 μg/L), with one system as the control treatment. Systems with 25 fish, on the other hand, were used as positive controls for acetylcholinesterase (10 μg/L of Atrazine) and oxidative stress (3 μg/L of Atrazine). After four months of exposure, a mean of 32 fish (S.D. ± 2) per group of MET treatment survived, which were used for the evaluation of behavior with the Novel Tank test. Moreover, after the behavioral assessment, we aimed to collect the blood and brains of all fish from all treatment groups. For blood collection, fish were anesthetized with an MS-222 solution (150 mg/L), while for brain gathering, fish were euthanized using the hypothermic shock method (2–4 °C). Blood was employed to determine CASP3 activity and the percentage of apoptotic cells with the TUNEL assay, and brains were used to evaluate acetylcholinesterase activity, oxidative damage, and gene expression. After chronic exposure, MET-exposed fish exhibited less swimming activity when compared to control fish. Moreover, compared with the control group, MET significantly inhibited the activity of AChE and induced oxidative damage in the brain of fish. Concerning gene expression, MET significantly upregulated the expression of Nrf1, Nrf2, BAX, p53, BACE1, APP, PSEN1, and downregulated CASP3 and CASP9. Although MET did not overexpress the CASP3 gene, we saw a meaningful rise in the activity of this enzyme in the blood of fish exposed to MET compared to the control group, which we then confirmed by a high number of apoptotic cells in the TUNEL assay. To the best of our understanding, this is the first study that delivers evidence of oxidative impairment, apoptosis, AChE alteration, and overexpression of B- amyloid-related genes in the brain of fish exposed to metformin.

Keywords: AChE inhibition, CASP3 activity, NovelTank test, oxidative damage, TUNEL assay

Procedia PDF Downloads 67
2226 Effect of Serine/Threonine Kinases on Autophagy Mechanism

Authors: Ozlem Oral, Seval Kilic, Ozlem Yedier, Serap Dokmeci, Devrim Gozuacik

Abstract:

Autophagy is a degradation pathway, activating under stress conditions. It digests macromolecules, such as abnormal proteins and long-lived organelles by engulfing them and by subsequent delivery of the cargo to lysosomes. The members of the phospholipid-dependent serine/threonine kinases, involved in many signaling pathways, which are necessary for the regulation of cellular metabolic activation. Previous studies implicate that, serine/threonine kinases have crucial roles in the mechanism of many diseases depend on the activated and/or inactivated signaling pathway. Data indicates, the signaling pathways activated by serine/threonine kinases are also involved in activation of autophagy mechanism. However, the information about the effect of serine/threonine kinases on autophagy mechanism and the roles of these effects in disease formation is limited. In this study, we investigated the effect of activated serine/threonine kinases on autophagic pathway. We performed a commonly used autophagy technique, GFP-LC3 dot formation and by using microscopy analyses, we evaluated promotion and/or inhibition of autophagy in serine/threonine kinase-overexpressed fibroblasts as well as cancer cells. In addition, we carried out confocal microscopy analyses and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. Based on the shRNA-library based screening, we identified autophagy-related proteins affected by serine/threonine kinases. We further studied the involvement of serine/threonine kinases on the molecular mechanism of newly identified autophagy proteins and found that, autophagic pathway is indirectly controlled by serine/threonine kinases via specific autophagic proteins. Our data indicate the molecular connection between two critical cellular mechanisms, which have important roles in the formation of many disease pathologies, particularly cancer. This project is supported by TUBITAK-1001-Scientific and Technological Research Projects Funding Program, Project No: 114Z836.

Keywords: autophagy, GFP-LC3 dot formation assay, serine/threonine kinases, shRNA-library screening

Procedia PDF Downloads 271
2225 Curcumin Nanomedicine: A Breakthrough Approach for Enhanced Lung Cancer Therapy

Authors: Shiva Shakori Poshteh

Abstract:

Lung cancer is a highly prevalent and devastating disease, representing a significant global health concern with profound implications for healthcare systems and society. Its high incidence, mortality rates, and late-stage diagnosis contribute to its formidable nature. To address these challenges, nanoparticle-based drug delivery has emerged as a promising therapeutic strategy. Curcumin (CUR), a natural compound derived from turmeric, has garnered attention as a potential nanomedicine for lung cancer treatment. Nanoparticle formulations of CUR offer several advantages, including improved drug delivery efficiency, enhanced stability, controlled release kinetics, and targeted delivery to lung cancer cells. CUR exhibits a diverse array of effects on cancer cells. It induces apoptosis by upregulating pro-apoptotic proteins, such as Bax and Bak, and downregulating anti-apoptotic proteins, such as Bcl-2. Additionally, CUR inhibits cell proliferation by modulating key signaling pathways involved in cancer progression. It suppresses the PI3K/Akt pathway, crucial for cell survival and growth, and attenuates the mTOR pathway, which regulates protein synthesis and cell proliferation. CUR also interferes with the MAPK pathway, which controls cell proliferation and survival, and modulates the Wnt/β-catenin pathway, which plays a role in cell proliferation and tumor development. Moreover, CUR exhibits potent antioxidant activity, reducing oxidative stress and protecting cells from DNA damage. Utilizing CUR as a standalone treatment is limited by poor bioavailability, lack of targeting, and degradation susceptibility. Nanoparticle-based delivery systems can overcome these challenges. They enhance CUR’s bioavailability, protect it from degradation, and improve absorption. Further, Nanoparticles enable targeted delivery to lung cancer cells through surface modifications or ligand-based targeting, ensuring sustained release of CUR to prolong therapeutic effects, reduce administration frequency, and facilitate penetration through the tumor microenvironment, thereby enhancing CUR’s access to cancer cells. Thus, nanoparticle-based CUR delivery systems promise to improve lung cancer treatment outcomes. This article provides an overview of lung cancer, explores CUR nanoparticles as a treatment approach, discusses the benefits and challenges of nanoparticle-based drug delivery, and highlights prospects for CUR nanoparticles in lung cancer treatment. Future research aims to optimize these delivery systems for improved efficacy and patient prognosis in lung cancer.

Keywords: lung cancer, curcumin, nanomedicine, nanoparticle-based drug delivery

Procedia PDF Downloads 51
2224 Crossbite Unilateral Correction Using Transpalatal Arch with Extension Arm Modification

Authors: Hanifa Maryani Ahmad, Muslim Yusuf

Abstract:

Background: Unilateral crossbite can be defined as an abnormal transverse relationship between the upper and lower teeth where the mandibular buccal cusp occluding to the maxillary buccal cusp and which involves only one side of the arch. This report describes the treatment of an adolescent female with Class III malocclussion unilateral crossbite resulting from a mildly constricted maxillary arch. The patient had a Class III skeletal relationship, Class III molar relationships, unilateral crossbite on the left side, and deviated midlines. Objectives: The treatment objectives were to correct the abnormal transverse relationship, achieve proper dental inclination, and correct the unilateral crossbites to improve the facial profile. Case management: The treatment protocol was using transpalatal arch with extension arm modification to expand the maxillary arch. Following the levelling and aligning stage of treatment, using a vertical loop while mandibular arch was expanded after getting an end to end relationship on the anterior side. Results: Corrections of the unilateral crossbite were achieved in 4 months. The treatment is still on process because the canines relationship were not corrected. Conclusions: This report highlights a treatment using transpalatal arch with extension arm modification that can be used to expand the transverse width of an arch to correct the discrepancy. Even though the treatment processes were still ongoing, the correction of the unilateral crossbite have been achieved in 4 months by only using the transpalatal arch.

Keywords: crossbite unilateral, late growing, non-extraction, transpalatal arch

Procedia PDF Downloads 194
2223 Biochemical Studies on the Effects of Cymbopogon citratus (Lemon Grass) on Wistar Albino Rats

Authors: Adegbegi Ademuyiwa Joshua, Onoagbe Iyare

Abstract:

Medicinal plants have been recognized to have therapeutic effects and they may also have toxic side effects. The present study was undertaken to investigate the effect of extracts of Cymbopogon citratus on normal rats. Blood glucose levels of all animals were determined. Biochemical studies carried out to determine the oxidative status by measuring activities of superoxide dismutase (SOD) and catalase (CAT), and in the liver, kidney and pancrease. Oral administration of ethanolic and aqueous extract of C. citratus at a doses of 200 mg/kg body weight, for a period of 30 days, caused a significant (p<0.05) reduction in blood glucose levels. Effect on hormonal profile (TSH, T3, and T4) was also determined, and was found to be significantly higher in all the administered groups when compared with control. Lipid profiles levels; Total cholesterols, triglycerides, high density lipoprotein-cholesterol and low density lipoprotein-cholesterol were significantly (p>0.05) higher for all treated rats as compared against control. SOD, catalase, GSH and Vitamin C activities in the tissues (liver, kidney and pancrease) of the rats treated with the medicinal plants were generally higher or statistical slightly similar to control. Histopathology result showed that both ethanolic and aqueous extracts (200 mg/kg body weight) of C. citratus was safer as no adverse effects were observed in the organs examined. Findings in this study showed that this plant has hypoglycemic properties and did not exert oxidative damage; in some instances, particularly in the liver, kidney and pancreas as well as its relative safety and possible use for weight gain.

Keywords: medicinal plants, blood glucose, cymbopogon citratus, hypoglycaemic, oxidative status

Procedia PDF Downloads 453
2222 The Molecule Preserve Environment: Effects of Inhibitor of the Angiotensin Converting Enzyme on Reproductive Potential and Composition Contents of the Mediterranean Flour Moth, Ephestia kuehniella Zeller

Authors: Yezli-Touiker Samira, Amrani-Kirane Leila, Soltani Mazouni Nadia

Abstract:

Due to secondary effects of conventional insecticides on the environment, the agrochemical research has resulted in the discovery of novel molecules. That research work will help in the development of a new group of pesticides that may be cheaper and less hazardous to the environment and non-target organisms which is the main desired outcome of the present work. Angiotensin-converting enzyme as a target for the development of novel insect growth regulators. Captopril is an inhibitor of angiotensin converting enzyme (ACE) it was tested in vivo by topical application on reproduction of Ephestia kuehniella Zeller (Lepidoptera: Pyralidae). The compound is diluted in acetone and applied topically to newly emerged pupae (10µg/ 2µl). The effects of this molecule was studied,on the biochemistry of ovary (on amounts nucleic acid, proteins, the qualitative analysis of the ovarian proteins and the reproductive potential (duration of the pre-oviposition, duration of the oviposition, number of eggs laid and hatching percentage). Captopril reduces significantly quantity of ovarian proteins and nucleic acid. The electrophoresis profile reveals the absence of tree bands at the treated series. This molecule reduced the duration of the oviposition period, the fecundity and the eggviability.

Keywords: environment, ephestia kuehniella, captopril, reproduction, the agrochemical research

Procedia PDF Downloads 262
2221 Design and in Slico Study of the Truncated Spike-M-N SARS-CoV-2 as a Novel Effective Vaccine Candidate

Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.

Abstract:

Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.

Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine

Procedia PDF Downloads 198
2220 Antibody Reactivity of Synthetic Peptides Belonging to Proteins Encoded by Genes Located in Mycobacterium tuberculosis-Specific Genomic Regions of Differences

Authors: Abu Salim Mustafa

Abstract:

The comparisons of mycobacterial genomes have identified several Mycobacterium tuberculosis-specific genomic regions that are absent in other mycobacteria and are known as regions of differences. Due to M. tuberculosis-specificity, the peptides encoded by these regions could be useful in the specific diagnosis of tuberculosis. To explore this possibility, overlapping synthetic peptides corresponding to 39 proteins predicted to be encoded by genes present in regions of differences were tested for antibody-reactivity with sera from tuberculosis patients and healthy subjects. The results identified four immunodominant peptides corresponding to four different proteins, with three of the peptides showing significantly stronger antibody reactivity and rate of positivity with sera from tuberculosis patients than healthy subjects. The fourth peptide was recognized equally well by the sera of tuberculosis patients as well as healthy subjects. Predication of antibody epitopes by bioinformatics analyses using ABCpred server predicted multiple linear epitopes in each peptide. Furthermore, peptide sequence analysis for sequence identity using BLAST suggested M. tuberculosis-specificity for the three peptides that had preferential reactivity with sera from tuberculosis patients, but the peptide with equal reactivity with sera of TB patients and healthy subjects showed significant identity with sequences present in nob-tuberculous mycobacteria. The three identified M. tuberculosis-specific immunodominant peptides may be useful in the serological diagnosis of tuberculosis.

Keywords: genomic regions of differences, Mycobacterium tuberculossis, peptides, serodiagnosis

Procedia PDF Downloads 164
2219 Using Scanning Electron Microscope and Computed Tomography for Concrete Diagnostics of Airfield Pavements

Authors: M. Linek

Abstract:

This article presents the comparison of selected evaluation methods regarding microstructure modification of hardened cement concrete intended for airfield pavements. Basic test results were presented for two pavement quality concrete lots. Analysis included standard concrete used for airfield pavements and modern material solutions based on concrete composite modification. In case of basic grain size distribution of concrete cement CEM I 42,5HSR NA, fine aggregate and coarse aggregate fractions in the form of granite chippings, water and admixtures were considered. In case of grain size distribution of modified concrete, the use of modern modifier as substitute of fine aggregate was suggested. Modification influence on internal concrete structure parameters using scanning electron microscope was defined. Obtained images were compared to the results obtained using computed tomography. Opportunity to use this type of equipment for internal concrete structure diagnostics and an attempt of its parameters evaluation was presented. Obtained test results enabled to reach a conclusion that both methods can be applied for pavement quality concrete diagnostics, with particular purpose of airfield pavements.

Keywords: scanning electron microscope, computed tomography, cement concrete, airfield pavements

Procedia PDF Downloads 314
2218 Modification of Polymer Composite Based on Electromagnetic Radiation

Authors: Ananta R. Adhikari

Abstract:

In today's era, polymer composite utilization has witnessed a significant increase across various fronts of material science advancement. Despite the development of many highly sophisticated technologies aimed at modifying polymer composites, there persists a quest for a technology that is straightforward, energy-efficient, easily controllable, cost-effective, time-saving, and environmentally friendly. Microwave technology has emerged as a major technique in material synthesis and modification due to its unique characteristics such as rapid, selective, uniform heating, and, particularly, direct heating based on molecular interaction. This study will be about the utilization of microwave energy as an alternative technique for material processing. Specifically, we will explore ongoing research conducted in our laboratory, focusing on its applications in the medical field.

Keywords: polymer composites, material processing, microstructure, microwave radiation

Procedia PDF Downloads 22
2217 Production of Recombinant VP2 Protein of Canine Parvovirus 2a Using Baculovirus Expression System

Authors: Soo Dong Cho, In-Ohk Ouh, Byeong Sul Kang, Seyeon Park, In-Soo Cho, Jae Young Song

Abstract:

An VP2 gene from the current prevalent CPV (Canine Parvovirus) strain (new CPV-2a) in the Republic of Korea was expressed in a baculovirus expression system. Genomic DNA was extracted from the isolate strain CPV-2a. The recombinant baculovirus, containing the coding sequences of VP2 with the histidine tag at the N-terminus, were generated by using the Bac-to-Bac system. For production of the recombinant VP2 proteins, SF9 cells were transfection into 6 wells. Propagation of recombinant baculoviruses and expression of the VP2 protein were performed in the Sf9 cell line maintained. The proteins were detected to Western blot anlaysis. CPV-2a VP2 was detected by Western blotting the monoclonal antibodies recognized 6x His and the band had a molecular weight of 65 KDa. We demonstrated that recombinant CPV-2a VP2 expression in baculovirus. The recombinant CPV-2a VP2 may able to development of specific diagnostic test and vaccination of against CPV2. This study provides a foundation for application of CPV2 on the development of new CPV2 subunit vaccine.

Keywords: baculovirus, canine parvovirus 2a, Dog, Korea

Procedia PDF Downloads 220
2216 Hepatoxicity induced Glyphosate-Based Herbicide Baron in albino rats

Authors: Manal E. A Elhalwagy, Nadia Amin Abdulmajeed, Hanan S. Alnahdi, Enas N. Danial

Abstract:

Baron is herbicide includes (48% glyphosate) widely used in Egypt. The present study assesses the cytotoxic and genotoxic effect of baron on rats liver. Two groups of rats were treated orally with 1/10 LD 50, (275.49 mg kg -1) and 1/40 LD 50, (68.86 mg kg-1) glyphosate for 28 days compared with control group. Serum and liver tissues were taken at 14 and 28 days of treatment. An inhibition in Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were recorded at both treatment periods and reduction in total serum protein (TP) and albumin (ALB). However, non-significant changes in serum acetylcholinesterase (AChE). Elevation in oxidative stress biomarker malondyaldehyde (MDA) and the decline in detoxification biomarker total reduced glutathione (GSH), Glutathione S-transferase (GST) and superoxide dismutase (SOD) in liver tissues led to increase in percentage of DNA damage. Destruction in liver tissue architecture was observed . Although, Baron was classified in the safe category pesticides repeated exposure to small doses has great danger effect.

Keywords: glyphosate, liver toxicity, oxidative stress, DNA damage, commet assay

Procedia PDF Downloads 354
2215 Establishment of Farmed Fish Welfare Biomarkers Using an Omics Approach

Authors: Pedro M. Rodrigues, Claudia Raposo, Denise Schrama, Marco Cerqueira

Abstract:

Farmed fish welfare is a very recent concept, widely discussed among the scientific community. Consumers’ interest regarding farmed animal welfare standards has significantly increased in the last years posing a huge challenge to producers in order to maintain an equilibrium between good welfare principles and productivity, while simultaneously achieve public acceptance. The major bottleneck of standard aquaculture is to impair considerably fish welfare throughout the production cycle and with this, the quality of fish protein. Welfare assessment in farmed fish is undertaken through the evaluation of fish stress responses. Primary and secondary stress responses include release of cortisol and glucose and lactate to the blood stream, respectively, which are currently the most commonly used indicators of stress exposure. However, the reliability of these indicators is highly dubious, due to a high variability of fish responses to an acute stress and the adaptation of the animal to a repetitive chronic stress. Our objective is to use comparative proteomics to identify and validate a fingerprint of proteins that can present an more reliable alternative to the already established welfare indicators. In this way, the culture conditions will improve and there will be a higher perception of mechanisms and metabolic pathway involved in the produced organism’s welfare. Due to its high economical importance in Portuguese aquaculture Gilthead seabream will be the elected species for this study. Protein extracts from Gilthead Seabream fish muscle, liver and plasma, reared for a 3 month period under optimized culture conditions (control) and induced stress conditions (Handling, high densities, and Hipoxia) are collected and used to identify a putative fish welfare protein markers fingerprint using a proteomics approach. Three tanks per condition and 3 biological replicates per tank are used for each analisys. Briefly, proteins from target tissue/fluid are extracted using standard established protocols. Protein extracts are then separated using 2D-DIGE (Difference gel electrophoresis). Proteins differentially expressed between control and induced stress conditions will be identified by mass spectrometry (LC-Ms/Ms) using NCBInr (taxonomic level - Actinopterygii) databank and Mascot search engine. The statistical analysis is performed using the R software environment, having used a one-tailed Mann-Whitney U-test (p < 0.05) to assess which proteins were differentially expressed in a statistically significant way. Validation of these proteins will be done by comparison of the RT-qPCR (Quantitative reverse transcription polymerase chain reaction) expressed genes pattern with the proteomic profile. Cortisol, glucose, and lactate are also measured in order to confirm or refute the reliability of these indicators. The identified liver proteins under handling and high densities induced stress conditions are responsible and involved in several metabolic pathways like primary metabolism (i.e. glycolysis, gluconeogenesis), ammonia metabolism, cytoskeleton proteins, signalizing proteins, lipid transport. Validition of these proteins as well as identical analysis in muscle and plasma are underway. Proteomics is a promising high-throughput technique that can be successfully applied to identify putative welfare protein biomarkers in farmed fish.

Keywords: aquaculture, fish welfare, proteomics, welfare biomarkers

Procedia PDF Downloads 122