Search results for: protein oxidation
2185 Metformin Protects Cardiac Muscle against the Pro-Apoptotic Effects of Hyperglycaemia, Elevated Fatty Acid and Nicotine
Authors: Christopher R. Triggle, Hong Ding, Khaled Machaca, Gnanapragasam Arunachalam
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The antidiabetic drug, metformin, has been in clinical use for over 50 years and remains the first choice drug for the treatment of type two diabetes. In addition to its effectiveness as an oral anti-hyperglycaemic drug metformin also possesses vasculoprotective effects that are assumed to be secondary to its ability to reduce insulin resistance and control glycated hemoglobin levels; however, recent data from our laboratory indicate that metformin also has direct vasoprotective effects that are mediated, at least in part, via the anti-ageing gene, SIRT1. Diabetes is a major risk factor for the development of cardiovascular disease (CVD) and it is also well established that tobacco use further enhances the risk of CVD; however, it is not known whether treatment with metformin can offset the negative effects of diabetes and tobacco use on cardiac function. The current study was therefore designed to investigate 1: the effects of hyperglycaemia (HG) either alone or in the presence of elevated fatty acids (palmitate) and nicotine on the protein expression levels of the deacetylase sirtuin 1 (the protein product of SIRT1), anti-apoptotic Bcl-2, pro-apoptotic BIM and the pro-apoptotic, tumour suppressor protein, acetylated p53 in cardiomyocytes. 2: the ability of metformin to prevent the detrimental effects of HG, palmitate and nicotine on cardiomyocyte survival. Cell culture protocols were designed using a rat cardiomyocyte cell line, H9c2, either under normal glycaemic (NG) conditions of 5.5mM glucose, or hyperglycaemic conditions (HG) of 25mM glucose with, or without, added palmitate (250μM) or nicotine (1.0mM) for 24h. Immuno-blotting was used to detect the expression of sirtuin 1, Bcl-2, BIM, acetylated (Ac)-p53, p53 with β-actin used as the reference protein. Exposure to HG, palmitate, or nicotine alone significantly reduced expression of sirtuin1, Bcl-2 and raised the expression levels of acetylated p53 and BIM; however, the combination of HG, palmitate and nicotine had a synergistic effect to significantly suppress the expression levels of sirtuin 1 and Bcl-2, but further enhanced the expression of Ac-p53, and BIM. The inclusion of 1000μM, but not 50μM, metformin in the H9c2 cell culture protocol prevented the effects of HG, palmitate and nicotine on the pro-apoptotic pathways. Collectively these data indicate that metformin, in addition to its anti-hyperglycaemic and vasculoprotective properties, also has direct cardioprotective actions that offset the negative effects of hyerglycaemia, elevated free fatty acids and nicotine on cardiac cell survival. These data are of particular significance for the treatment of patients with diabetes who are also smokers as the inclusion of metformin in their therapeutic treatment plan should help reduce cardiac-related morbidity and mortality.Keywords: apoptosis, cardiac muscle, diabetes, metformin, nicotine
Procedia PDF Downloads 3112184 Elevated Creatinine Clearance and Normal Glomerular Filtration Rate in Patients with Systemic Lupus erythematosus
Authors: Stoyanka Vladeva, Elena Kirilova, Nikola Kirilov
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Background: The creatinine clearance is a widely used value to estimate the GFR. Increased creatinine clearance is often called hyperfiltration and is usually seen during pregnancy, patients with diabetes mellitus preceding the diabetic nephropathy. It may also occur with large dietary protein intake or with plasma volume expansion. Renal injury in lupus nephritis is known to affect the glomerular, tubulointerstitial, and vascular compartment. However high creatinine clearance has not been found in patients with SLE, Target: Follow-up of creatinine clearance values in patients with systemic lupus erythematosus without history of kidney injury. Material and methods: We observed the creatinine, creatinine clearance, GFR and dipstick protein values of 7 women (with a mean age of 42.71 years) with systemic lupus erythematosus. Patients with active lupus have been monthly tested in the period of 13 months. Creatinine clearance has been estimated by Cockcroft-Gault Equation formula in ml/sec. GFR has been estimated by MDRD formula (The Modification of Diet in renal Disease) in ml/min/1.73 m2. Proteinuria has been defined as present when dipstick protein > 1+.Results: In all patients without history of kidney injury we found elevated creatinine clearance levels, but GFRremained within the reference range. Two of the patients were in remission while the other five patients had clinically and immunologically active Lupus. Three of the patients had a permanent presence of high creatinine clearance levels and proteinuria. Two of the patients had periodically elevated creatinine clearance without proteinuria. These results show that kidney disturbances may be caused by the vascular changes typical for SLE. Glomerular hyperfiltration can be result of focal segmental glomerulosclerosis caused by a reduction in renal mass. Probably lupus nephropathy is preceded not only by glomerular vascular changes, but also by tubular vascular changes. Using only the GFR is not a sufficient method to detect these primary functional disturbances. Conclusion: For early detection of kidney injury in patients with SLE we determined that the follow up of creatinine clearance values could be helpful.Keywords: systemic Lupus erythematosus, kidney injury, elevated creatinine clearance level, normal glomerular filtration rate
Procedia PDF Downloads 2662183 In vivo Determination of Anticoagulant Property of the Tentacle Extract of Aurelia aurita (Moon Jellyfish) Using Sprague-Dawley Rats
Authors: Bea Carmel H. Casiding, Charmaine A. Guy, Funny Jovis P. Malasan, Katrina Chelsea B. Manlutac, Danielle Ann N. Novilla, Marianne R. Oliveros, Magnolia C. Sibulo
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Moon jellyfish, Aurelia aurita, has become a popular research organism for diverse studies. Recent studies have verified the prevention of blood clotting properties of the moon jellyfish tentacle extract through in vitro methods. The purpose of this study was to validate the blood clotting ability of A. aurita tentacle extract using in vivo method of experimentation. The tentacles of A. aurita jellyfish were excised and filtered then centrifuged at 3000xg for 10 minutes. The crude nematocyst extract was suspended in 1:6 ratios with phosphate buffer solution and sonicated for three periods of 20 seconds each at 50 Hz. Protein concentration of the extract was determined using Bradford Assay. Bovine serum albumin was the standard solution used with the following concentrations: 35.0, 70.0, 105.0, 140.0, 175.0, 210.0, 245.0, and 280.0 µg/mL. The absorbance was read at 595 nm. Toxicity testing from OECD guidelines was adapted. The extract suspended in phosphate-buffered saline solution was arbitrarily set into three doses (0.1mg/kg, 0.3mg/kg, 0.5mg/kg) and were administered daily for five days to the experimental groups of five male Sprague-Dawley rats (one dose per group). Before and after the administration period, bleeding time and clotting time tests were performed. The One-way Analysis of Variance (ANOVA) was used to analyze the difference of before and after bleeding time and clotting time from the three treatment groups, time, positive and negative control groups. The average protein concentration of the sonicated crude tentacle extract was 206.5 µg/mL. The highest dose administered (0.5mg/kg) produced significant increase in the time for both bleeding and clotting tests. However, the preceding lower dose (0.3mg/kg) only was significantly effective for clotting time test. The protein contained in the tentacle extract with a concentration of 206.5 mcg/mL and dose of 0.3 mg/kg and 0.5 mg/kg of A. aurita elicited anticoagulating activity.Keywords: anticoagulant, bleeding time test, clotting time test, moon jellyfish
Procedia PDF Downloads 3922182 Inhibition of Variant Surface Glycoproteins Translation to Define the Essential Features of the Variant Surface Glycoprotein in Trypanosoma brucei
Authors: Isobel Hambleton, Mark Carrington
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Trypanosoma brucei, the causal agent of a range of diseases in humans and livestock, evades the mammalian immune system through a population survival strategy based on the expression of a series of antigenically distinct variant surface glycoproteins (VSGs). RNAi mediated knockdown of the active VSG gene triggers a precytokinesis cell cycle arrest. To determine whether this phenotype is the result of reduced VSG transcript or depleted VSG protein, we used morpholino antisense oligonucleotides to block translation of VSG mRNA. The same precytokinesis cell cycle arrest was observed, suggesting that VSG protein abundance is monitored closely throughout the cell cycle. An inducible expression system has been developed to test various GPI-anchored proteins for their ability to rescue this cell cycle arrest. This system has been used to demonstrate that wild-type VSG expressed from a T7 promoter rescues this phenotype. This indicates that VSG expression from one of the specialised bloodstream expression sites (BES) is not essential for cell division. The same approach has been used to define the minimum essential features of a VSG necessary for function.Keywords: bloodstream expression site, morpholino, precytokinesis cell cycle arrest, variant surface glycoprotein
Procedia PDF Downloads 1432181 The Effects of Myelin Basic Protein Charge Isomers on the Methyl Cycle Metabolites in Glial Cells
Authors: Elene Zhuravliova, Tamar Barbakadze, Irina Kalandadze, Elnari Zaalishvili, Lali Shanshiashvili, David Mikeladze
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Background: Multiple sclerosis (MS) is an inflammatory, neurodegenerative disease, which is accompanied by demyelination and autoimmune response to myelin proteins. Among post-translational modifications, which mediate the modulation of inflammatory pathways during MS, methylation is the main one. The methylation of DNA, also amino acids lysine and arginine, occurs in the cell. It was found that decreased trans-methylation is associated with neuroinflammatory diseases. Therefore, abnormal regulation of the methyl cycle could induce demyelination through the action on PAD (peptidyl-arginine-deiminase) gene promoter. PAD takes part in protein citrullination and targets myelin basic protein (MBP), which is affected during demyelination. To determine whether MBP charge isomers are changing the methyl cycle, we have estimated the concentrations of methyl cycle metabolites in MBP-activated primary astrocytes and oligodendrocytes. For this purpose, the action of the citrullinated MBP- C8 and the most cationic MBP-C1 isomers on the primary cells were investigated. Methods: Primary oligodendrocyte and astrocyte cell cultures were prepared from whole brains of 2-day-old Wistar rats. The methyl cycle metabolites, including homocysteine, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH), were estimated by HPLC analysis using fluorescence detection and prior derivatization. Results: We found that the action of MBP-C8 and MBP-C1 induces a decrease in the concentration of both methyl cycle metabolites, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), in astrocytes compared to the control cells. As for oligodendrocytes, the concentration of SAM was increased by the addition of MBP-C1, while MBP-C8 has no significant effect. As for SAH, its concentration was increased compared to the control cells by the action of both MBP-C1 and MBP-C8. A significant increase in homocysteine concentration was observed by the action of the MBP-C8 isomer in both oligodendrocytes and astrocytes. Conclusion: These data suggest that MBP charge isomers change the concentration of methyl cycle metabolites. MBP-C8 citrullinated isomer causes elevation of homocysteine in astrocytes and oligodendrocytes, which may be the reason for decreased astrocyte proliferation and increased oligodendrocyte cell death which takes place in neurodegenerative processes. Elevated homocysteine levels and subsequent abnormal regulation of methyl cycles in oligodendrocytes possibly change the methylation of DNA that activates PAD gene promoter and induces the synthesis of PAD, which in turn provokes the process of citrullination, which is the accompanying process of demyelination. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.Keywords: myelin basic protein, astrocytes, methyl cycle metabolites, homocysteine, oligodendrocytes
Procedia PDF Downloads 1472180 Effect of Crude Flowers Extract of Citrus reticulata Blanco Flowers on Physicochemical and Nutritional Properties of Cheddar Cheese
Authors: Usman Mir Khan, Ishtiaque Ahmad, Saima Inayat, H. M. Arslan Amin, Muhammad Ayaz, Nisar Ahmad
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Citrus reticulata Blanco crude flowers extract (CFE) at four different concentration (1, 2, 3 and 4%, v/v) were used as natural milk coagulant instead of rennet to apply for Cheddar cheese making from buffalo milk. The physicochemical properties and nutrition composition of Cheddar cheeses were compared with cheese made with 0.002% (v/v) rennet (control cheese). Physico-chemical of Cheddar cheese showed that cheese made with 1% and 2% of CFE had a crumbly and slightly softer texture of cheese. While, cheeses containing 3 and 4% CFE had semi-hard textural properties of curd similar to rennet added cheese. The CFE made cheese had moisture 37 %, fat 45 % on dry basis similar to rennet made Cheddar cheese. Protein analysis shows that CFE made cheese had significant higher protein content than control. The Cheddar cheese with 3% and 1% CFE were preferred by consumers instead of 2% and 4% CFE for their taste, texture/appearance and overall acceptability. Conclusively, CFE coagulated Cheddar cheese fulfills the nutritional requirement with acceptable organoleptic characteristics and at the same time provides nutritional health benefits.Keywords: cheddar cheese, Citrus reticulata Blanco, buffalo milk, milk coagulant
Procedia PDF Downloads 2962179 A Comparative Study of Euglena gracilis Cultivations for Improving Laminaribiose Phosphorylase Production
Authors: Akram Abi, Clarissa Müller, Hans-Joachim Jördening
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Laminaribiose is a beta-1,3-glycoside which is used in the medical field for the treatment of dermatitis and also can be used as a building block for new pharmaceutics. The conventional process of laminaribiose production is the uneconomical process of hydrolysis of laminarin extracted from natural polysaccharides of plant origin. A more economical approach however is attainable by enzymatically synthesis of laminaribiose via a reverse phosphorylase reaction catalyzed by laminaribiose phosphorylase (LP) from Euglena gracilis. Different cultivation methods of Euglena gracilis and the effect on LP production have been investigated. Buffered/unbuffered heterotrophic and mixotrophic cultivations of Euglena gracilis has been carried out. Changes of biomass and LP production, glucose level and pH, cell count and shape has been monitored in the course of time. The results obtained from experiments each in three repetitions, show that in the heterotrophic cultivation of Euglena gracilis not only more biomass is produced compared to mixotrophic cultivation, but also higher specific protein concentration is achieved. Furthermore, the LP activity test showed that the protein extracted from heterotrophically cultured cells has a higher LP activity. It was also observed that the cells develop in a distinctive different shape between these two cultures and have different length to width ratios. Taking the heterotrophic culture as the more efficient cultivation method in LP production, another comparative experiment between buffered and unbuffered heterothrophic culture was carried out that showed the unbuffered culture has advantages over the other one in respect of both LP production and resulting activity. A hetrotrophic cultivation of Euglena gracilis in a 5L bioreactor with controlled operating conditions showed a distinctive improvement of all the aspects of culture compared to the shaking flask cultivations. Biomass production was improved from 5 to more than 8 g/l (dry weight) which resulted in a specific protein concentration of 45 g/l in the heterotrophic cultivation in the bioreactor. In further attempts to improve LP production, different purification methods were tested and each method was checks through an activity assay. A laminaribiose yield of 35% was achieved which was by far the highest amount amongst different methods tested.Keywords: euglena gracilis, heterotrophic culture, laminaribiose production, mixotrophic culture
Procedia PDF Downloads 3582178 Production of Recombinant Human Serum Albumin in Escherichia coli: A Crucial Biomolecule for Biotechnological and Healthcare Applications
Authors: Ashima Sharma, Tapan K. Chaudhuri
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Human Serum Albumin (HSA) is one of the most demanded therapeutic protein with immense biotechnological applications. The current source of HSA is human blood plasma. Blood is a limited and an unsafe source as it possesses the risk of contamination by various blood derived pathogens. This issue led to exploitation of various hosts with the aim to obtain an alternative source for the production of the rHSA. But, till now no host has been proven to be effective commercially for rHSA production because of their respective limitations. Thus, there exists an indispensable need to promote non-animal derived rHSA production. Of all the host systems, Escherichia coli is one of the most convenient hosts which has contributed in the production of more than 30% of the FDA approved recombinant pharmaceuticals. E. coli grows rapidly and its culture reaches high cell density using inexpensive and simple substrates. The fermentation batch turnaround number for E. coli culture is 300 per year, which is far greater than any of the host systems available. Therefore, E. coli derived recombinant products have more economical potential as fermentation processes are cheaper compared to the other expression hosts available. Despite of all the mentioned advantages, E. coli had not been successfully adopted as a host for rHSA production. The major bottleneck in exploiting E. coli as a host for rHSA production was aggregation i.e. majority of the expressed recombinant protein was forming inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA form inclusion body is not preferred because it is tedious, time consuming, laborious and expensive. Because of this limitation, E. coli host system was neglected for rHSA production for last few decades. Considering the advantages of E. coli as a host, the present work has targeted E. coli as an alternate host for rHSA production through resolving the major issue of inclusion body formation associated with it. In the present study, we have developed a novel and innovative method for enhanced soluble and functional production of rHSA in E.coli (~60% of the total expressed rHSA in the soluble fraction) through modulation of the cellular growth, folding and environmental parameters, thereby leading to significantly improved and enhanced -expression levels as well as the functional and soluble proportion of the total expressed rHSA in the cytosolic fraction of the host. Therefore, in the present case we have filled in the gap in the literature, by exploiting the most well studied host system Escherichia coli which is of low cost, fast growing, scalable and ‘yet neglected’, for the enhancement of functional production of HSA- one of the most crucial biomolecule for clinical and biotechnological applications.Keywords: enhanced functional production of rHSA in E. coli, recombinant human serum albumin, recombinant protein expression, recombinant protein processing
Procedia PDF Downloads 3392177 Cultivation And Production of Insects, Especially Mealworms (Mealworms) and Investigating Its Potential as Food for Animals and Even Humans
Authors: Marzieh Eshaghi Koupaei
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By cultivating mealworm, we reduce greenhouse gases and avoid the use of transgenic products such as soybeans, and we provide food resources rich in protein, amino acids, minerals, etc. for humans and animals, and it has created employment and entrepreneurship. We serve the environment by producing oil from mealworm in the cosmetic industry, using its waste as organic fertilizer and its powder in bodybuilding, and by breaking down plastic by mealworm. The production and breeding of mealworm requires very little infrastructure and does not require much trouble, and requires very little food, and reproduces easily and quickly, and a mealworm production workshop is noiseless, odorless, and pollution-free And the costs are very low. It is possible to use third grade fruits and unsalable fruits of farmers to feed the mealworms, which is completely economical and cost-effective. Mealworms can break down plastic in their intestines and turn it into carbon dioxide. . This process was done in only 16 days, which is a very short time compared to several centuries for plastic to decompose. By producing mealworm, we have helped to preserve the environment and provided the source of protein needed by humans and animals. This industrial insect has the ability and value of commercialization and creates employment and helps the economy of the society.Keywords: breeding, production of insects, mealworms, research, animal feed, human feed
Procedia PDF Downloads 462176 Astaxanthin Induces Cytotoxicity through Down-Regulating Rad51 Expression in Human Lung Cancer Cells
Authors: Jyh-Cheng Chen, Tai-Jing Wang, Yun-Wei Lin
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Astaxanthin has been demonstrated to exhibit a wide range of beneficial effects including anti-inflammatory and anti-cancer properties. However, the molecular mechanism of astaxanthin-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Rad51 plays a central role in homologous recombination and high levels of Rad51 expression are observed in chemo- or radioresistant carcinomas. In this study, astaxanthin treatment inhibited cell viability and proliferation of two NSCLC cells, A549 and H1703. Treatment with astaxanthin decreased Rad51 expression and phospho-AKT protein level in a time and dose-dependent manner. Furthermore, expression of constitutively active AKT (AKT-CA) vector significantly rescued the decreased Rad51 protein and mRNA levels in astaxanthin-treated NSCLC cells. Combined treatment with PI3K inhibitors (LY294002 or wortmannin) and astaxanthin further decreased the Rad51 expression in NSCLC cells. Knockdown of Rad51 enhanced astaxanthin-induced cytotoxicity and growth inhibition in NSCLC cells. These findings may have implications for the rational design of future drug regimens incorporating astaxanthin for the treatment of NSCLC.Keywords: astaxanthin, cytotoxicity, AKT, non-small cell lung cancer, PI3K
Procedia PDF Downloads 2902175 The Strategy for Detection of Catecholamines in Body Fluids: Optical Sensor
Authors: Joanna Cabaj, Sylwia Baluta, Karol Malecha, Kamila Drzozga
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Catecholamines are the principal neurotransmitters that mediate a variety of the central nervous system functions, such as motor control, cognition, emotion, memory processing, and endocrine modulation. Dysfunctions in catecholamine neurotransmission are induced in some neurologic and neuropsychiatric diseases. Changeable neurotransmitters level in biological fluids can be a marker of several neurological disorders. Because of its significance in analytical techniques and diagnostics, sensitive and selective detection of neurotransmitters is increasingly attracting a lot of attention in different areas of bio-analysis or biomedical research. Recently, fluorescent techniques for detection of catecholamines have attracted interests due to their reasonable cost, convenient control, as well as maneuverability in biological environments. Nevertheless, with the observed need for a sensitive and selective catecholamines sensor, the development of a convenient method for this neurotransmitter is still at its basic level. The manipulation of nanostructured materials in conjunction with biological molecules has led to the development of a new class of hybrid modified biosensors in which both enhancement of charge transport and biological activity preservation may be obtained. Immobilization of biomaterials on electrode surfaces is the crucial step in fabricating electrochemical as well as optical biosensors and bioelectronic devices. Continuing systematic investigation in the manufacturing of enzyme–conducting sensitive systems, here is presented a convenient fluorescence sensing strategy for catecholamines detection based on FRET (fluorescence resonance energy transfer) phenomena observed for, i.e., complexes of Fe²⁺ and epinephrine. The biosensor was constructed using low temperature co-fired ceramics technology (LTCC). This sensing system used the catalytical oxidation of catecholamines and quench of the strong luminescence of obtained complexes due to FRET. The detection process was based on the oxidation of substrate in the presence of the enzyme–laccase/tyrosinase.Keywords: biosensor, conducting polymer, enzyme, FRET, LTCC
Procedia PDF Downloads 2532174 Protein Derived Biodegradable Food Packaging Material from Poultry By-Product
Authors: Muhammad Zubair, Aman Ullah, Jianping Wu
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During the last decades, petroleum derived synthetic polymers like polyethylene terephthalate, polyvinylchloride, polyethylene, polypropylene and polystyrene has extensively been used in the field of food packaging and mostly are non-degradable. Biopolymers are a good fit for single-use or short-lived products such as food packaging. Spent hens, a poultry by-product which is of little economic value and their disposal are environmentally harmful. Through current study, we have explored the possibility to transform proteins from spent fowl into green food packaging material. Proteins from spent fowl were extracted within 1 hour using pH shift method with recovery of about 74%. Different plasticizers were tried like glycerol, sorbitol, glutaraldehyde, 1,2 ethylene glycol and 1,2 butanediol. Glycerol was the best plasticizer among all these plasticizers. A naturally occurring and non-toxic cross-linking agent, chitosan, was used to form the chitosan/glycerol/protein blend by casting and compression molding techniques. The mechanical properties were characterized using tensile strength analyzer. The nano-reinforcements with homogeneous dispersion of nanoparticles lead to improved physical properties suggesting that these materials have great potential for food packaging applications.Keywords: differential scanning calorimetry, dynamic mechanical analysis, scanning electron microscopy, spent hen
Procedia PDF Downloads 2722173 Efficacy of Microbial Metabolites Obtained from Saccharomyces cerevisiae as Supplement for Quality Milk Production in Dairy Cows
Authors: Sajjad ur Rahman, Mariam Azam, Mukarram Bashir, Seemal Javaid, Aoun Muhammad, Muhammad Tahir, Jawad, Hannan Khan, Muhammad Zohaib
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Partially fermented soya hulls and wheat bran through Saccharomyces cerevisiae (DL-22 S/N) substantiated as a natural source for quality milk production. Saccharomyces cerevisiae (DL-22 S/N) were grown under in-vivo conditions and processed through two-step fermentation with substrates. The extra pure metabolites (XPM) were dried and processed for maintaining 1mm mesh size particles for supplementation of pelleted feed. Two groups of a cow (Holstein Friesian) having 8 animals of similar age and lactation were given the experimental concentrates. Group A was fed daily with 12gm of XPM and 22% protein-pelleted feed, while Group B was provided with no metabolites in their feed. In thirty-nine days of trial, improvement in the overall health, body score, milk protein, milk fat, ash, and solid not fat (SNF), yield, and incidence rate of mastitis was observed. The collected data revealed an improvement in milk production of 2.02 liter/h/d. However, a reduction (3.75%) in the milk fats and an increase in the milk SNF was around 0.58%. The ash content ranged between 6.4-7.5%. The incidence of mastitis was reduced to less than 2%.Keywords: microbial metabolites, Saccharomyces cerevisiae, milk production, fermentation, post-biotic metabolites, immunity
Procedia PDF Downloads 812172 Carbon Nanotube-Based Catalyst Modification to Improve Proton Exchange Membrane Fuel Cell Interlayer Interactions
Authors: Ling Ai, Ziyu Zhao, Zeyu Zhou, Xiaochen Yang, Heng Zhai, Stuart Holmes
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Optimizing the catalyst layer structure is crucial for enhancing the performance of proton exchange membrane fuel cells (PEMFCs) with low Platinum (Pt) loading. Current works focused on the utilization, durability, and site activity of Pt particles on support, and performance enhancement has been achieved by loading Pt onto porous support with different morphology, such as graphene, carbon fiber, and carbon black. Some schemes have also incorporated cost considerations to achieve lower Pt loading. However, the design of the catalyst layer (CL) structure in the membrane electrode assembly (MEA) must consider the interactions between the layers. Addressing the crucial aspects of water management, low contact resistance, and the establishment of effective three-phase boundary for MEA, multi-walled carbon nanotubes (MWCNTs) are promising CL support due to their intrinsically high hydrophobicity, high axial electrical conductivity, and potential for ordered alignment. However, the drawbacks of MWCNTs, such as strong agglomeration, wall surface chemical inertness, and unopened ends, are unfavorable for Pt nanoparticle loading, which is detrimental to MEA processing and leads to inhomogeneous CL surfaces. This further deteriorates the utilization of Pt and increases the contact resistance. Robust chemical oxidation or nitrogen doping can introduce polar functional groups onto the surface of MWCNTs, facilitating the creation of open tube ends and inducing defects in tube walls. This improves dispersibility and load capacity but reduces length and conductivity. Consequently, a trade-off exists between maintaining the intrinsic properties and the degree of functionalization of MWCNTs. In this work, MWCNTs were modified based on the operational requirements of the MEA from the viewpoint of interlayer interactions, including the search for the optimal degree of oxidation, N-doping, and micro-arrangement. MWCNT were functionalized by oxidizing, N-doping, as well as micro-alignment to achieve lower contact resistance between CL and proton exchange membrane (PEM), better hydrophobicity, and enhanced performance. Furthermore, this work expects to construct a more continuously distributed three-phase boundary by aligning MWCNT to form a locally ordered structure, which is essential for the efficient utilization of Pt active sites. Different from other chemical oxidation schemes that used HNO3:H2SO4 (1:3) mixed acid to strongly oxidize MWCNT, this scheme adopted pure HNO3 to partially oxidize MWCNT at a lower reflux temperature (80 ℃) and a shorter treatment time (0 to 10 h) to preserve the morphology and intrinsic conductivity of MWCNT. The maximum power density of 979.81 mw cm-2 was achieved by Pt loading on 6h MWCNT oxidation time (Pt-MWCNT6h). This represented a 59.53% improvement over the commercial Pt/C catalyst of 614.17 (mw cm-2). In addition, due to the stronger electrical conductivity, the charge transfer resistance of Pt-MWCNT6h in the electrochemical impedance spectroscopy (EIS) test was 0.09 Ohm cm-2, which was 48.86% lower than that of Pt/C. This study will discuss the developed catalysts and their efficacy in a working fuel cell system. This research will validate the impact of low-functionalization modification of MWCNTs on the performance of PEMFC, which simplifies the preparation challenges of CL and contributing for the widespread commercial application of PEMFCs on a larger scale.Keywords: carbon nanotubes, electrocatalyst, membrane electrode assembly, proton exchange membrane fuel cell
Procedia PDF Downloads 582171 Oat Grain Functional Ingredient Characterization
Authors: Vita Sterna, Sanita Zute, Inga Jansone, Linda Brunava, Inara Kantane
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Grains, including oats (Avena sativa L.), have been recognized functional foods, because provide beneficial effect on the health of the consumer and decrease the risk of various diseases.Oats are good source of soluble fibre, essential amino acids, unsaturated fatty acids, vitamins and minerals. Oat breeders have developed oat varieties and improved yielding ability potential of oat varieties. Therefore, the aim of investigation was to analyze the composition of perspective oat varieties and breeding lines grains grown in different conditions and evaluate functional properties. In the studied samples content of protein, starch, β - glucans, total dietetic fibre, composition of amino acids and vitamin E were determined. The results of analysis showed that protein content depending of varieties ranged 9.70 –17.30% total dietary fibre 13.66-30.17 g100g-1, content of β-glucans 2.7-3.5 g100g-1, amount of vitamin E (α-tocopherol) determined from 4 to 9.9 mg kg-1. The sum of essential amino acids in oat grain samples were determined from 31.63 to 54.90 gkg-1. Concluded that amino acids composition of husked and naked oats grown in organic or conventional conditions is close to optimal.Keywords: dietetic fibre, amino acids, scores, nutrition value
Procedia PDF Downloads 4942170 Electrochemical Bioassay for Haptoglobin Quantification: Application in Bovine Mastitis Diagnosis
Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo
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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine
Procedia PDF Downloads 1632169 Influence of Salicylic Acid on Yield and Some Physiological Parameters in Chickpea (Cicer arietinum L.)
Authors: Farid Shekari
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Salicylic Acid (SA) is a plant hormone that improves some physiological responses of plants under stress conditions. Seeds of two desi type chickpea cultivars, viz., Kaka and Pirooz, primed with 250, 500, 750, and 1000 μM of SA and a group of seeds without any treating (as control) were evaluated under rain fed conditions. Seed priming in both cultivars led to higher efficiency compare to non-primed treatments. In general, seed priming with 500 and 750 μM of SA had appropriate effects; however the cultivars responses were different in this regard. Kaka showed better performance both in primed and non-primed seed than Pirooz. Results of this study revealed that not only yield quantity but also yield quality, as seed protein amounts, could positively affect by SA treatments. It seems that SA by enhancing of soluble sugars and proline amounts enhanced total water potential (ψ) and RWC. The increment in RWC led to rose of chlorophyll content of plants chlorophyll stability. In general, SA increased water use efficiency, both in biologic and seed yield base, and drought tolerance of chickpea plants. HI was a little decreased in SA treatments and it shows that SA more effective in biomass production than seed yield.Keywords: chlorophyll, harvest index, proline, seed protein, soluble sugar, water use efficiency, yield component
Procedia PDF Downloads 4162168 Effects of Raw Bee Propolis and Water or Ethanol Extract of Propolis on Performance, Immune System and Some Blood Parameters on Broiler Bredeers
Authors: Hasan Alp Sahin, Ergin Ozturk
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The effects of raw bee propolis (RP) and water (WEP) or ethanol (EEP) extract of propolis on growth performance, selected immune parameters (IgA, IgY and IgM) and some blood parameters such as aspartate aminotransferase, alanine aminotransferase, trygliceride, total protein, albumin, calcium, phosphorus, total antioxidant status and total oxidant status were determined. The study was conducted between 15th and 20th weeks (6 weeks) and used a total of 48 broiler breeder pullets (Ross-308). The broiler breeder in control group was fed diet without propolis whereas the birds in RP, WEP and EEP groups were fed diets with RP, WEP and EEP at the level of 1200, 400 and 400 ppm, respectively. All pullets were fed mash form diet with 15% crude protein and 2800 ME kcal/kg. All propolis forms had not a beneficial effect on any studied parameters compared to control group (P > 0.05). The results of the study indicated that both the level of the active matters supplied from the bee propolis has no enough beneficial effect on performance, some immune and blood parameters on broiler breeders or they did not have such a level that would cause a beneficial effect on these variables.Keywords: antioxidant, bee product , poultry breeders, growth performance, immune parameters, blood chemistry
Procedia PDF Downloads 2562167 Mannosidase Alpha Class 1B Member 1 Targets F Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein and Ebola Virus Glycoprotein to Endoplasmic Reticulum-To-Lysosome-Associated Degradation by Micro-Endoplasmic Reticulum-Phagy
Authors: Yong-Hui Zheng
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Viruses hijack host machineries to propagate and spread, which disrupts cellular homeostasis and activates various counteractive mechanisms. Infection of enveloped viruses is dependent on their fusion proteins, which bind to viral receptors to allow virus entry into cells. Fusion proteins are glycoproteins and expressed in the endoplasmic reticulum (ER) by hijacking the secretory pathway. Previously, we reported that Zaire ebolavirus (EBOV)-glycoprotein (GP) expression induces ER stress, and EBOV-GP is targeted by the calnexin cycle to macro-ER-phagy for degradation. We now report that expression of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2/SARS2)-spike (S) protein also causes ER stress, and its expression is strongly downregulated by mannosidase alpha class 1B member 1 (MAN1B1), a class I α-mannosidase from the ER. MAN1B1 co-localizes with SARS2-S in the ER, and its downregulation of SARS2-S is blocked by inhibitors targeting lysosomes and autophagy, but not proteasomes, indicating SARS2-S degradation by autolysosomes. Notably, the SARS2-S degradation does not require the core autophagy machinery including ATG3, ATG5, ATG7, and phosphatidylinositol 3-kinase catalytic subunit type 3 (PI3KC3)/vacuolar protein sorting 34 (VPS34), and instead, it requires Beclin 1 (BECN1), a core component in the PI3KC3 complex. In addition, MAN1B1 does not trigger SARS2-S polyubiquitination, and consistently, the SARS2-S degradation does not require the autophagy receptor sequestosome 1 (SQSTM1)/p62. MAN1B1 also downregulates EBOV-GP similarly, but this degradation does not require BECN1. Collectively, we conclude that MAN1B1 downregulates viral fusions by micro-ER-phagy, and importantly, we have identified BECN1-dependent and BECN1-independent mechanisms for micro-ER-phagy.Keywords: Micro-ER-phagy, reticulophagy, fusion proteins, ER stress
Procedia PDF Downloads 612166 Degradation of Emerging Pharmaceuticals by Gamma Irradiation Process
Authors: W. Jahouach-Rabai, J. Aribi, Z. Azzouz-Berriche, R. Lahsni, F. Hosni
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Gamma irradiation applied in removing pharmaceutical contaminants from wastewater is an effective advanced oxidation process (AOP), considered as an alternative to conventional water treatment technologies. In this purpose, the degradation efficiency of several detected contaminants under gamma irradiation was evaluated. In fact, radiolysis of organic pollutants in aqueous solutions produces powerful reactive species, essentially hydroxyl radical ( ·OH), able to destroy recalcitrant pollutants in water. Pharmaceuticals considered in this study are aqueous solutions of paracetamol, ibuprofen, and diclofenac at different concentrations 0.1-1 mmol/L, which were treated with irradiation doses from 3 to 15 kGy. The catalytic oxidation of these compounds by gamma irradiation was investigated using hydrogen peroxide (H₂O₂) as a convenient oxidant. Optimization of the main parameters influencing irradiation process, namely irradiation doses, initial concentration and oxidant volume (H₂O₂) were investigated, in the aim to release high degradation efficiency of considered pharmaceuticals. Significant modifications attributed to these parameters appeared in the variation of degradation efficiency, chemical oxygen demand removal (COD) and concentration of radio-induced radicals, confirming them synergistic effect to attempt total mineralization. Pseudo-first-order reaction kinetics could be used to depict the degradation process of these compounds. A sophisticated analytical study was released to quantify the detected radio-induced radicals (electron paramagnetic resonance spectroscopy (EPR) and high performance liquid chromatography (HPLC)). All results showed that this process is effective for the degradation of many pharmaceutical products in aqueous solutions due to strong oxidative properties of generated radicals mainly hydroxyl radical. Furthermore, the addition of an optimal amount of H₂O₂ was efficient to improve the oxidative degradation and contribute to the high performance of this process at very low doses (0.5 and 1 kGy).Keywords: AOP, COD, hydroxyl radical, EPR, gamma irradiation, HPLC, pharmaceuticals
Procedia PDF Downloads 1672165 Sardine Oil as a Source of Lipid in the Diet of Giant Freshwater Prawn (Macrobrachium rosenbergii)
Authors: A. T. Ramachandra Naik, H. Shivananda Murthy, H. n. Anjanayappa
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The freshwater prawn, Macrobrachium rosenbergii is a more popular crustacean cultured widely in monoculture system in India. It has got high nutritional value in the human diet. Hence, understanding its enzymatic and body composition is important in order to judge its flesh quality. Fish oil specially derived from Indian oil sardine is a good source of highly unsaturated fatty acid and lipid source in fish/prawn diet. A 35% crude protein diet with graded levels of Sardine oil as a source of fat was incorporated at four levels viz, 2.07, 4.07, 6.07 and 8.07% maintaining a total lipid level of feed at 8.11, 10.24, 12.28 and 14.33% respectively. Diet without sardine oil (6.05% total lipid) was served as basal treatment. The giant freshwater prawn, Macrobrachium rosenbergii was used as test animal and the experiment was lost for 112 days. Significantly, higher gain in weight of prawn was recorded in the treatment with 6.07% sardine oil incorporation followed by higher specific growth rate, food conversion rate and protein efficiency ratio. The 8.07% sardine oil diet produced the highest RNA: DNA ratio in the prawn muscle. Digestive enzyme analyses in the digestive tract and mid-gut gland showed the greatest activity in prawns fed the 8.07% diet.Keywords: digestive enzyme, fish diet, Macrobrachium rosenbergii, sardine oil
Procedia PDF Downloads 3202164 Effects of Pterostilbene in Brown Adipose Tissue from Obese Rats
Authors: Leixuri Aguirre, Iñaki Milton-Laskibar, Elizabeth Hijona, Luis Bujanda, Agnes M. Rimando, Maria P. Portillo
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Introduction: In recent years great attention has been paid by scientific community to phenolic compounds as active biomolecules naturally present in foodstuffs due to their beneficial effects on health. Pterostilbene is a resveratrol dimethylether derivative which shows higher biodisponibility. Objective. To analyze the effects of two doses of pterostilbene on several markers of thermogenic capacity in a model of genetic obesity, which shows reduced thermogenesis. Methods: The experiment was conducted with thirty Zucker (fa/fa) rats that were distributed in 3 experimental groups, the control group and two groups orally administered with pterostilbene at 15 and 30 mg/kg body weight/day for 6 weeks. Gene expression of Ucp1, Pgc-1α, Cpt1b, Pparα, Nfr1, Tfam and Cox-2 were assessed by RT-PCR, protein expression of UCP1 and GLUT4 by western blot and enzyme activity of carnitine palmitoyl transferase 1b and citrate synthase by spectrophotometry in interscapular brown adipose tissue (iBAT). Statistical analysis was performed by using one way ANOVA and Newman-Keuls as post-hoc test. Results: Pterostilbene did not change gene expression of Pgc-1α. However, significant increases were found in the expression of Ucp1, Pparα, Nfr-1 and Cox-2. Protein expression of UCP1 and GLUT4 was increased in animals treated with pterostilbene, as well as the activities of CPT-1b and CS. These effects were observed with both doses of pterostilbene, without differences between them. Conclusions: These results show that pterostilbene increases thermogenic and oxidative capacity of brown adipose tissue in obese rats. Whether these effects effectively contribute to the anti-obesity properties of these compound needs further research. Acknowledgments: MINECO-FEDER (AGL2015-65719-R), Basque Government (IT-572-13), University of the Basque Country (ELDUNANOTEK UFI11/32), Institut of Health Carlos III (CIBERobn). Iñaki Milton is a fellowship from the Basque Government.Keywords: brown adipose tissue, pterostilbene, thermogenesis, uncoupling protein 1
Procedia PDF Downloads 2892163 The Nutritive Value of Fermented Sago Pith (Metroxylon sago Rottb) Enriched with Micro Nutrients for Poultry Feed
Authors: Wizna, Helmi Muis, Hafil Abbas
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An experiment was conducted to improve the nutrient value of sago pith (Metroxylon sago Rottb) supplemented with Zn, Sulfur and urea through fermentation by using cellulolytic bacteria (Bacillus amyloliquefaciens) as inoculums. The experiment was determination of the optimum dose combination (dosage of Zn, S and urea) for sago pith fermentation based on nutrient quality and quantity of these fermented products. The study was conducted in experimental method, using the completely randomized design in factorial with 3 treatments consist of: factor A (Dose of urea: A1 = 2.0%, A2 = 3.0%), factor B (Dose of S: B1 = 0.2%, B2 = 0.4%) and factor C (Dose of Zn: C1 = 0.0025%, C2 = 0.005%). Results of study showed that optimum condition for fermentation process of sago pith with B. amyloliquefaciens caused a change of nutrient content was obtained at urea (3%), S (0,2%), and Zn (0,0025%). This fermentation process was able to increase amino acid average, reduce crude fiber content by 67% and increase crude protein by 433%, which made the nutritional value of the product based on dry matter was 18.22% crude protein, 12.42% crude fiber, 2525 Kcal/kg metabolic energy and 65.73% nitrogen retention.Keywords: fermentation, sago pith, sulfur, Zn, urea, Bacillus amyloliquefaciens
Procedia PDF Downloads 5042162 Dynamic Cardiac Mitochondrial Proteome Alterations after Ischemic Preconditioning
Authors: Abdelbary Prince, Said Moussa, Hyungkyu Kim, Eman Gouda, Jin Han
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We compared the dynamic alterations of mitochondrial proteome of control, ischemia-reperfusion (IR) and ischemic preconditioned (IPC) rabbit hearts. Using 2-DE, we identified 29 mitochondrial proteins that were differentially expressed in the IR heart compared with the control and IPC hearts. For two of the spots, the expression patterns were confirmed by Western blotting analysis. These proteins included succinate dehydrogenase complex, Acyl-CoA dehydrogenase, carnitine acetyltransferase, dihydrolipoamide dehydrogenase, Atpase, ATP synthase, dihydrolipoamide succinyltransferase, ubiquinol-cytochrome c reductase, translation elongation factor, acyl-CoA dehydrogenase, actin alpha, succinyl-CoA Ligase, dihydrolipoamide S-succinyltransferase, citrate synthase, acetyl-Coenzyme A dehydrogenase, creatine kinase, isocitrate dehydrogenase, pyruvate dehydrogenase, prohibitin, NADH dehydrogenase (ubiquinone) Fe-S protein, enoyl Coenzyme A hydratase, superoxide dismutase [Mn], and 24-kDa subunit of complex I. Interestingly, most of these proteins are associated with the mitochondrial respiratory chain, antioxidant enzyme system, and energy metabolism. The results provide clues as to the cardioprotective mechanism of ischemic preconditioning at the protein level and may serve as potential biomarkers for detection of ischemia-induced cardiac injury.Keywords: ischemic preconditioning, mitochondria, proteome, cardioprotection
Procedia PDF Downloads 3442161 Applying Computer Simulation Methods to a Molecular Understanding of Flaviviruses Proteins towards Differential Serological Diagnostics and Therapeutic Intervention
Authors: Sergio Alejandro Cuevas, Catherine Etchebest, Fernando Luis Barroso Da Silva
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The flavivirus genus has several organisms responsible for generating various diseases in humans. Special in Brazil, Zika (ZIKV), Dengue (DENV) and Yellow Fever (YFV) viruses have raised great health concerns due to the high number of cases affecting the area during the last years. Diagnostic is still a difficult issue since the clinical symptoms are highly similar. The understanding of their common structural/dynamical and biomolecular interactions features and differences might suggest alternative strategies towards differential serological diagnostics and therapeutic intervention. Due to their immunogenicity, the primary focus of this study was on the ZIKV, DENV and YFV non-structural proteins 1 (NS1) protein. By means of computational studies, we calculated the main physical chemical properties of this protein from different strains that are directly responsible for the biomolecular interactions and, therefore, can be related to the differential infectivity of the strains. We also mapped the electrostatic differences at both the sequence and structural levels for the strains from Uganda to Brazil that could suggest possible molecular mechanisms for the increase of the virulence of ZIKV. It is interesting to note that despite the small changes in the protein sequence due to the high sequence identity among the studied strains, the electrostatic properties are strongly impacted by the pH which also impact on their biomolecular interactions with partners and, consequently, the molecular viral biology. African and Asian strains are distinguishable. Exploring the interfaces used by NS1 to self-associate in different oligomeric states, and to interact with membranes and the antibody, we could map the strategy used by the ZIKV during its evolutionary process. This indicates possible molecular mechanisms that can explain the different immunological response. By the comparison with the known antibody structure available for the West Nile virus, we demonstrated that the antibody would have difficulties to neutralize the NS1 from the Brazilian strain. The present study also opens up perspectives to computationally design high specificity antibodies.Keywords: zika, biomolecular interactions, electrostatic interactions, molecular mechanisms
Procedia PDF Downloads 1232160 Mutations in the GJB2 Gene Are the Cause of an Important Number of Non-Syndromic Deafness Cases
Authors: Habib Onsori, Somayeh Akrami, Mohammad Rahmati
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Deafness is the most common sensory disorder with the frequency of 1/1000 in many populations. Mutations in the GJB2 (CX26) gene at the DFNB1 locus on chromosome 13q12 are associated with congenital hearing loss. Approximately 80% of congenital hearing loss cases are recessively inherited and 15% dominantly inherited. Mutations of the GJB2 gene, encoding gap junction protein Connexin 26 (Cx26), are the most common cause of hereditary congenital hearing loss in many countries. This report presents two cases of different mutations from Iranian patients with bilateral hearing loss. DNA studies were performed for the GJB2 gene by PCR and sequencing methods. In one of them, direct sequencing of the gene showed a heterozygous T→C transition at nucleotide 604 resulting in a cysteine to arginine amino acid substitution at codon 202 (C202R) in the fourth extracellular domain (TM4) of the protein. The analyses indicate that the C202R mutation appeared de novo in the proband with a possible dominant effect (GenBank: KF 638275). In the other one, DNA sequencing revealed a compound heterozygous mutation (35delG, 363delC) in the Cx26 gene that is strongly associated with congenital non-syndromic hearing loss (NSHL). So screening the mutations for hearing loss individuals referring to genetics counseling centers before marriage and or pregnancy is recommended.Keywords: CX26, deafness, GJB2, mutation
Procedia PDF Downloads 4792159 Comparative Proteomic Profiling of Planktonic and Biofilms from Staphylococcus aureus Using Tandem Mass Tag-Based Mass Spectrometry
Authors: Arifur Rahman, Ardeshir Amirkhani, Honghua Hu, Mark Molloy, Karen Vickery
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Introduction and Objectives: Staphylococcus aureus and coagulase-negative staphylococci comprises approximately 65% of infections associated with medical devices and are well known for their biofilm formatting ability. Biofilm-related infections are extremely difficult to eradicate owing to their high tolerance to antibiotics and host immune defences. Currently, there is no efficient method for early biofilm detection. A better understanding to enable detection of biofilm specific proteins in vitro and in vivo can be achieved by studying planktonic and different growth phases of biofilms using a proteome analysis approach. Our goal was to construct a reference map of planktonic and biofilm associated proteins of S. aureus. Methods: S. aureus reference strain (ATCC 25923) was used to grow 24 hours planktonic, 3-day wet biofilm (3DWB), and 12-day wet biofilm (12DWB). Bacteria were grown in tryptic soy broth (TSB) liquid medium. Planktonic growth was used late logarithmic bacteria, and the Centres for Disease Control (CDC) biofilm reactor was used to grow 3 days, and 12-day hydrated biofilms, respectively. Samples were subjected to reduction, alkylation and digestion steps prior to Multiplex labelling using Tandem Mass Tag (TMT) 10-plex reagent (Thermo Fisher Scientific). The labelled samples were pooled and fractionated by high pH RP-HPLC which followed by loading of the fractions on a nanoflow UPLC system (Eksigent UPLC system, AB SCIEX). Mass spectrometry (MS) data were collected on an Orbitrap Elite (Thermo Fisher Scientific) Mass Spectrometer. Protein identification and relative quantitation of protein levels were performed using Proteome Discoverer (version 1.3, Thermo Fisher Scientific). After the extraction of protein ratios with Proteome Discoverer, additional processing, and statistical analysis was done using the TMTPrePro R package. Results and Discussion: The present study showed that a considerable proteomic difference exists among planktonic and biofilms from S. aureus. We identified 1636 total extracellular secreted proteins, of which 350 and 137 proteins of 3DWB and 12DWB showed significant abundance variation from planktonic preparation, respectively. Of these, simultaneous up-regulation in between 3DWB and 12DWB proteins such as extracellular matrix-binding protein ebh, enolase, transketolase, triosephosphate isomerase, chaperonin, peptidase, pyruvate kinase, hydrolase, aminotransferase, ribosomal protein, acetyl-CoA acetyltransferase, DNA gyrase subunit A, glycine glycyltransferase and others we found in this biofilm producer. On the contrary, simultaneous down-regulation in between 3DWB and 12DWB proteins such as alpha and delta-hemolysin, lipoteichoic acid synthase, enterotoxin I, serine protease, lipase, clumping factor B, regulatory protein Spx, phosphoglucomutase, and others also we found in this biofilm producer. In addition, we also identified a big percentage of hypothetical proteins including unique proteins. Therefore, a comprehensive knowledge of planktonic and biofilm associated proteins identified by S. aureus will provide a basis for future studies on the development of vaccines and diagnostic biomarkers. Conclusions: In this study, we constructed an initial reference map of planktonic and various growth phase of biofilm associated proteins which might be helpful to diagnose biofilm associated infections.Keywords: bacterial biofilms, CDC bioreactor, S. aureus, mass spectrometry, TMT
Procedia PDF Downloads 1642158 The Effects of Different Agroforestry Practices on Glomalin Related Soil Protein, Soil Aggregate Stability and Organic Carbon-Association with Soil Aggregates in Southern Ethiopia
Authors: Nebiyou Masebo
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The severities of land degradation in southern Ethiopia has been increasing due to high population density, replacement of an age-old agroforestry (AF) based agricultural system with monocropping. The consequences of these activities combined with climate change have been impaired soil biota, soil organic carbon (SOC), soil glomalin, soil aggregation and aggregate stability. The AF systems could curb these problems due it is an ecologically and economically sustainable. This study was aimed to determine the effect of agroforestry practices (AFPs) on soil glomalin, soil aggregate stability (SAS), and aggregate association with SOC. Soil samples (from two depth level: 0-30 & 30-60 cm) and woody species were collected from homegarden based agroforestry practice (HAFP), cropland based agroforestry practice (ClAFP), woodlot based agroforestry practice (WlAFP) and trees on soil and water conservation based agroforestry practice (TSWAFP) using systematic sampling. In this study, both easily extractable glomalin related soil protein (EEGRSP) and total glomalin related soil protein (TGRSP) were significantly (p<0.05) higher in HAFP compared to others, with decreasing order HAFP>WlAFP>TSWAFP>ClAFP at upper surface but in subsurface in decreasing order: WlAFP>HAFP>TSWAFP>ClAFP. On the other hand, the macroaggregate fraction of AFPs ranged from 22.64-36.51% where the lowest was in ClAFP, while the highest was in HAFP, moreover, the order for subsurface was also the same but SAS decreased with the increasing of soil depths. The micro-aggregate fraction ranged from 15.9–24.56%, where the lowest was in HAFP, but the highest was in ClAFP. Besides, the association of OC with both macro-and micro-aggregates was greatest in HAFP and followed by WlAFP. The findings also showed that both glomalin and SAS were significantly high with woody species diversity and richness. Thus, AFP with good management practice can play role on maintenance of biodiversity, glomalin content and other soil quality parameters with future implications for a stable ecosystem.Keywords: agroforestry, soil aggregate stability, glomalin, aggregate-associated carbon, HAFP, ClAFP, WlAFP, TSWAFP.
Procedia PDF Downloads 932157 Femtochemistry of Iron(III) Carboxylates in Aqueous Solutions
Authors: Ivan P. Pozdnyakov, Alexey A. Melnikov, Nikolai V. Tkachenko
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Photochemical reactions with participation of iron (III) carboxylates are important for environmental photochemistry and have a great potential of application in water purification (Advanced Oxidation Processes, photo-Fenton and Fenton-like processes). In spite of this information about excited states and primary intermediates in photochemistry of Fe(III) complexes with carboxylic acids is scarce. This talk presents and discusses the results of several recent authors' publications in a field of ultra fast spectroscopy of natural Fe(III) carboxylates.Keywords: carboxylates, iron complexes, photochemistry, radical complexes, ultrafast processes
Procedia PDF Downloads 4522156 Electrochemical Inactivation of Toxic Cyanobacteria and Degradation of Cyanotoxins
Authors: Belal Bakheet, John Beardall, Xiwang Zhang, David McCarthy
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The potential risks associated with toxic cyanobacteria have raised growing environmental and public health concerns leading to an increasing effort into researching ways to bring about their removal from water, together with destruction of their associated cyanotoxins. A variety of toxins are synthesized by cyanobacteria and include hepatotoxins, neurotoxins, and cytotoxins which can cause a range of symptoms in humans from skin irritation to serious liver and nerve damage. Therefore drinking water treatment processes should ensure the consumers’ safety by removing both cyanobacterial cells, and cyanotoxins from the water. Cyanobacterial cells and cyanotoxins presented challenges to the conventional water treatment systems; their accumulation within drinking water treatment plants has been reported leading to plants shut down. Thus, innovative and effective water purification systems to tackle cyanobacterial pollution are required. In recent years there has been increasing attention to the electrochemical oxidation process as a feasible alternative disinfection method which is able to generate in situ a variety of oxidants that would achieve synergistic effects in the water disinfection process and toxin degradation. By utilizing only electric current, the electrochemical process through electrolysis can produce reactive oxygen species such as hydroxyl radicals from the water, or other oxidants such as chlorine from chloride ions present in the water. From extensive physiological and morphological investigation of cyanobacterial cells during electrolysis, our results show that these oxidants have significant impact on cell inactivation, simultaneously with cyanotoxins removal without the need for chemicals addition. Our research aimed to optimize existing electrochemical oxidation systems and develop new systems to treat water containing toxic cyanobacteria and cyanotoxins. The research covers detailed mechanism study on oxidants production and cell inactivation in the treatment under environmental conditions. Overall, our study suggests that the electrochemical treatment process e is an effective method for removal of toxic cyanobacteria and cyanotoxins.Keywords: toxic cyanobacteria, cyanotoxins, electrochemical process, oxidants
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