Search results for: core protein
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4142

Search results for: core protein

3212 Protein Derived Biodegradable Food Packaging Material from Poultry By-Product

Authors: Muhammad Zubair, Aman Ullah, Jianping Wu

Abstract:

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

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3211 Detailed Depositional Resolutions in Upper Miocene Sands of HT-3X Well, Nam Con Son Basin, Vietnam

Authors: Vo Thi Hai Quan

Abstract:

Nam Con Son sedimentary basin is one of the very important oil and gas basins in offshore Vietnam. Hai Thach field of block 05-2 contains mostly gas accumulations in fine-grained, sand/mud-rich turbidite system, which was deposited in a turbidite channel and fan environment. Major Upper Miocene reservoir of HT-3X lies above a well-developed unconformity. The main objectives of this study are to reconstruct depositional environment and to assess the reservoir quality using data from 14 meters of core samples and digital wireline data of the well HT-3X. The wireline log and core data showed that the vertical sequences of representative facies of the well mainly range from Tb to Te divisions of Bouma sequences with predominance of Tb and Tc compared to Td and Te divisions. Sediments in this well were deposited in a submarine fan association with very fine to fine-grained, homogeneous sandstones that have high porosity and permeability, high- density turbidity currents with longer transport route from the sediment source to the basin, indicating good quality of reservoir. Sediments are comprised mainly of the following sedimentary structures: massive, laminated sandstones, convoluted bedding, laminated ripples, cross-laminated ripples, deformed sandstones, contorted bedding.

Keywords: Hai Thach field, Miocene sand, turbidite, wireline data

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3210 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

Abstract:

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

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3209 Facility Data Model as Integration and Interoperability Platform

Authors: Nikola Tomasevic, Marko Batic, Sanja Vranes

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Emerging Semantic Web technologies can be seen as the next step in evolution of the intelligent facility management systems. Particularly, this considers increased usage of open source and/or standardized concepts for data classification and semantic interpretation. To deliver such facility management systems, providing the comprehensive integration and interoperability platform in from of the facility data model is a prerequisite. In this paper, one of the possible modelling approaches to provide such integrative facility data model which was based on the ontology modelling concept was presented. Complete ontology development process, starting from the input data acquisition, ontology concepts definition and finally ontology concepts population, was described. At the beginning, the core facility ontology was developed representing the generic facility infrastructure comprised of the common facility concepts relevant from the facility management perspective. To develop the data model of a specific facility infrastructure, first extension and then population of the core facility ontology was performed. For the development of the full-blown facility data models, Malpensa and Fiumicino airports in Italy, two major European air-traffic hubs, were chosen as a test-bed platform. Furthermore, the way how these ontology models supported the integration and interoperability of the overall airport energy management system was analyzed as well.

Keywords: airport ontology, energy management, facility data model, ontology modeling

Procedia PDF Downloads 426
3208 The Immunology Evolutionary Relationship between Signal Transducer and Activator of Transcription Genes from Three Different Shrimp Species in Response to White Spot Syndrome Virus Infection

Authors: T. C. C. Soo, S. Bhassu

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Unlike the common presence of both innate and adaptive immunity in vertebrates, crustaceans, in particular, shrimps, have been discovered to possess only innate immunity. This further emphasizes the importance of innate immunity within shrimps in pathogenic resistance. Under the study of pathogenic immune challenge, different shrimp species actually exhibit varying degrees of immune resistance towards the same pathogen. Furthermore, even within the same shrimp species, different batches of challenged shrimps can have different strengths of immune defence. Several important pathways are activated within shrimps during pathogenic infection. One of them is JAK-STAT pathway that is activated during bacterial, viral and fungal infections by which STAT(Signal Transducer and Activator of Transcription) gene is the core element of the pathway. Based on theory of Central Dogma, the genomic information is transmitted in the order of DNA, RNA and protein. This study is focused in uncovering the important evolutionary patterns present within the DNA (non-coding region) and RNA (coding region). The three shrimp species involved are Macrobrachium rosenbergii, Penaeus monodon and Litopenaeus vannamei which all possess commercial significance. The shrimp species were challenged with a famous penaeid shrimp virus called white spot syndrome virus (WSSV) which can cause serious lethality. Tissue samples were collected during time intervals of 0h, 3h, 6h, 12h, 24h, 36h and 48h. The DNA and RNA samples were then extracted using conventional kits from the hepatopancreas tissue samples. PCR technique together with designed STAT gene conserved primers were utilized for identification of the STAT coding sequences using RNA-converted cDNA samples and subsequent characterization using various bioinformatics approaches including Ramachandran plot, ProtParam and SWISS-MODEL. The varying levels of immune STAT gene activation for the three shrimp species during WSSV infection were confirmed using qRT-PCR technique. For one sample, three biological replicates with three technical replicates each were used for qRT-PCR. On the other hand, DNA samples were important for uncovering the structural variations within the genomic region of STAT gene which would greatly assist in understanding the STAT protein functional variations. The partially-overlapping primers technique was used for the genomic region sequencing. The evolutionary inferences and event predictions were then conducted through the Bayesian Inference method using all the acquired coding and non-coding sequences. This was supplemented by the construction of conventional phylogenetic trees using Maximum likelihood method. The results showed that adaptive evolution caused STAT gene sequence mutations between different shrimp species which led to evolutionary divergence event. Subsequently, the divergent sites were correlated to the differing expressions of STAT gene. Ultimately, this study assists in knowing the shrimp species innate immune variability and selection of disease resistant shrimps for breeding purpose. The deeper understanding of STAT gene evolution from the perspective of both purifying and adaptive approaches not only can provide better immunological insight among shrimp species, but also can be used as a good reference for immunological studies in humans or other model organisms.

Keywords: gene evolution, JAK-STAT pathway, immunology, STAT gene

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3207 Oat Grain Functional Ingredient Characterization

Authors: Vita Sterna, Sanita Zute, Inga Jansone, Linda Brunava, Inara Kantane

Abstract:

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

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3206 Electrochemical Bioassay for Haptoglobin Quantification: Application in Bovine Mastitis Diagnosis

Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

Abstract:

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

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3205 Development of Thermal Regulating Textile Material Consisted of Macrocapsulated Phase Change Material

Authors: Surini Duthika Fernandopulle, Kalamba Arachchige Pramodya Wijesinghe

Abstract:

Macrocapsules containing phase change material (PCM) PEG4000 as core and Calcium Alginate as the shell was synthesized by in-situ polymerization process, and their suitability for textile applications was studied. PCM macro-capsules were sandwiched between two polyurethane foams at regular intervals, and the sandwiched foams were subsequently covered with 100% cotton woven fabrics. According to the mathematical modelling and calculations 46 capsules were required to provide cooling for a period of 2 hours at 56ºC, so a panel of 10 cm x 10 cm area with 25 parts (having 5 capsules in each for 9 parts are 16 parts spaced for air permeability) were effectively merged into one textile material without changing the textile's original properties. First, the available cooling techniques related to textiles were considered and the best cooling techniques suiting the Sri Lankan climatic conditions were selected using a survey conducted for Sri Lankan Public based on ASHRAE-55-2010 standard and it consisted of 19 questions under 3 sections categorized as general information, thermal comfort sensation and requirement of Personal Cooling Garments (PCG). The results indicated that during daytime, majority of respondents feel warm and during nighttime also majority have responded as slightly warm. The survey also revealed that around 85% of the respondents are willing to accept a PCG. The developed panels were characterized using Fourier-transform infrared spectroscopy (FTIR) and Thermogravimetric Analysis (TGA) tests and the findings from FTIR showed that the macrocapsules consisted of PEG 4000 as the core material and Calcium Alginate as the shell material and findings from TGA showed that the capsules had the average weight percentage for core with 61,9% and shell with 34,7%. After heating both control samples and samples incorporating PCM panels, it was discovered that only the temperature of the control sample increased after 56ºC, whereas the temperature of the sample incorporating PCM panels began to regulate the temperature at 56ºC, preventing a temperature increase beyond 56ºC.

Keywords: phase change materials, thermal regulation, textiles, macrocapsules

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3204 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

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3203 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

Abstract:

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

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3202 Reliability Indices Evaluation of SEIG Rotor Core Magnetization with Minimum Capacitive Excitation for WECs

Authors: Lokesh Varshney, R. K. Saket

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This paper presents reliability indices evaluation of the rotor core magnetization of the induction motor operated as a self-excited induction generator by using probability distribution approach and Monte Carlo simulation. Parallel capacitors with calculated minimum capacitive value across the terminals of the induction motor operating as a SEIG with unregulated shaft speed have been connected during the experimental study. A three phase, 4 poles, 50Hz, 5.5 hp, 12.3A, 230V induction motor coupled with DC Shunt Motor was tested in the electrical machine laboratory with variable reactive loads. Based on this experimental study, it is possible to choose a reliable induction machine operating as a SEIG for unregulated renewable energy application in remote area or where grid is not available. Failure density function, cumulative failure distribution function, survivor function, hazard model, probability of success and probability of failure for reliability evaluation of the three phase induction motor operating as a SEIG have been presented graphically in this paper.

Keywords: residual magnetism, magnetization curve, induction motor, self excited induction generator, probability distribution, Monte Carlo simulation

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3201 Advances in Fiber Optic Technology for High-Speed Data Transmission

Authors: Salim Yusif

Abstract:

Fiber optic technology has revolutionized telecommunications and data transmission, providing unmatched speed, bandwidth, and reliability. This paper presents the latest advancements in fiber optic technology, focusing on innovations in fiber materials, transmission techniques, and network architectures that enhance the performance of high-speed data transmission systems. Key advancements include the development of ultra-low-loss optical fibers, multi-core fibers, advanced modulation formats, and the integration of fiber optics into next-generation network architectures such as Software-Defined Networking (SDN) and Network Function Virtualization (NFV). Additionally, recent developments in fiber optic sensors are discussed, extending the utility of optical fibers beyond data transmission. Through comprehensive analysis and experimental validation, this research offers valuable insights into the future directions of fiber optic technology, highlighting its potential to drive innovation across various industries.

Keywords: fiber optics, high-speed data transmission, ultra-low-loss optical fibers, multi-core fibers, modulation formats, coherent detection, software-defined networking, network function virtualization, fiber optic sensors

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3200 Teacher Professional Development Programs on K-12 Engineering Education: A Systematic Review of the Literature

Authors: Canan Mesutoglu, Evrim Baran

Abstract:

Teachers have a prominent role in facilitating the place of engineering in K-12 classrooms. This study addresses the need to understand how teacher professional development programs focusing on K-12 engineering education can be designed and delivered more effectively. A systematic review of the literature on such programs can offer possible ideas and recommendations. The purpose of this study is to systematically synthesize the peer-reviewed articles published on K-12 engineering education teacher professional development programs. The methodology that guided the study was comprised of four phases: search, selection, coding, and synthesis. The search phase included articles published in the time period between 2000 and 2016. With a comprehensive search in databases, inclusion criteria were applied. This was followed by evaluation of the quality of articles with a checklist, and finally analysis of the results. The results revealed two categories of themes. These were 1) five themes related to the overarching agenda of the PD programs, and 2) five themes related to the instructional techniques of the PD programs. Finally, core elements were generated to guide the design and delivery of teacher PD programs for K-12 engineering education. The results aimed to provide a conceptual basis for future research and practice on teacher PD programs for K-12 engineering education.

Keywords: core elements, K-12 engineering education, systematic literature review, teacher professional development programs

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3199 Creating Energy Sustainability in an Enterprise

Authors: John Lamb, Robert Epstein, Vasundhara L. Bhupathi, Sanjeev Kumar Marimekala

Abstract:

As we enter the new era of Artificial Intelligence (AI) and Cloud Computing, we mostly rely on the Machine and Natural Language Processing capabilities of AI, and Energy Efficient Hardware and Software Devices in almost every industry sector. In these industry sectors, much emphasis is on developing new and innovative methods for producing and conserving energy and sustaining the depletion of natural resources. The core pillars of sustainability are economic, environmental, and social, which is also informally referred to as the 3 P's (People, Planet and Profits). The 3 P's play a vital role in creating a core Sustainability Model in the Enterprise. Natural resources are continually being depleted, so there is more focus and growing demand for renewable energy. With this growing demand, there is also a growing concern in many industries on how to reduce carbon emissions and conserve natural resources while adopting sustainability in corporate business models and policies. In our paper, we would like to discuss the driving forces such as Climate changes, Natural Disasters, Pandemic, Disruptive Technologies, Corporate Policies, Scaled Business Models and Emerging social media and AI platforms that influence the 3 main pillars of Sustainability (3P’s). Through this paper, we would like to bring an overall perspective on enterprise strategies and the primary focus on bringing cultural shifts in adapting energy-efficient operational models. Overall, many industries across the globe are incorporating core sustainability principles such as reducing energy costs, reducing greenhouse gas (GHG) emissions, reducing waste and increasing recycling, adopting advanced monitoring and metering infrastructure, reducing server footprint and compute resources (Shared IT services, Cloud computing, and Application Modernization) with the vision for a sustainable environment.

Keywords: climate change, pandemic, disruptive technology, government policies, business model, machine learning and natural language processing, AI, social media platform, cloud computing, advanced monitoring, metering infrastructure

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3198 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

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3197 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

Abstract:

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

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3196 The Nutritive Value of Fermented Sago Pith (Metroxylon sago Rottb) Enriched with Micro Nutrients for Poultry Feed

Authors: Wizna, Helmi Muis, Hafil Abbas

Abstract:

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

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3195 Revealing the Structural and Dynamic Properties of Betaine Aldehyde Dehydrogenase 2 from Rice (Oryza sativa): Simulation Studies

Authors: Apisaraporn Baicharoen, Prapasiri Pongprayoon

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Betaine aldehyde dehydrogenase 2 (BADH2) is an enzyme that inhibits the accumulation of 2-acetyl-1-pyrroline (2AP), a potent flavor compound in rice fragrance. BADH2 contains three domains (NAD-binding, substrate-binding, and oligomerization domains). It catalyzes the oxidation of amino aldehydes. The lack of BADH2 results in the formation of 2AP and consequently an increase in rice fragrance. To date, inadequate data on BADH2 structure and function are available. An insight into the nature of BADH2 can serve as one of key starting points for the production of high quality fragrant rice. In this study, we therefore constructed the homology model of BADH2 and employed 500-ns Molecular Dynamics simulations (MD) to primarily understand the structural and dynamic properties of BADH2. Initially, Ramachandran plot confirms the good quality of modeled protein structure. Principle Component Analysis (PCA) was also calculated to capture the protein dynamics. Among 3 domains, the results show that NAD binding site is found to be more flexible. Moreover, interactions from key amino acids (N162, E260, C294, and Y419) that are crucial for function are investigated.

Keywords: betaine aldehyde dehydrogenase 2, fragrant rice, homology modeling, molecular dynamics simulations

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3194 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

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3193 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

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3192 Mutations in the GJB2 Gene Are the Cause of an Important Number of Non-Syndromic Deafness Cases

Authors: Habib Onsori, Somayeh Akrami, Mohammad Rahmati

Abstract:

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

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3191 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

Abstract:

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

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3190 Analysis of Combined Heat Transfer through the Core Materials of VIPs with Various Scattering Properties

Authors: Jaehyug Lee, Tae-Ho Song

Abstract:

Vacuum insulation panel (VIP) can achieve very low thermal conductivity by evacuating its inner space. Heat transfer in the core materials of highly-evacuated VIP occurs by conduction through the solid structure and radiation through the pore. The effect of various scattering modes in combined conduction-radiation in VIP is investigated through numerical analysis. The discrete ordinates interpolation method (DOIM) incorporated with the commercial code FLUENT® is employed. It is found that backward scattering is more effective in reducing the total heat transfer while isotropic scattering is almost identical with pure absorbing/emitting case of the same optical thickness. For a purely scattering medium, the results agree well with additive solution with diffusion approximation, while a modified term is added in the effect of optical thickness to backward scattering is employed. For other scattering phase functions, it is also confirmed that backwardly scattering phase function gives a lower effective thermal conductivity. Thus, the materials with backward scattering properties, with radiation shields are desirable to lower the thermal conductivity of VIPs.

Keywords: combined conduction and radiation, discrete ordinates interpolation method, scattering phase function, vacuum insulation panel

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3189 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

Abstract:

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.

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3188 Designing and Implementation of MPLS Based VPN

Authors: Muhammad Kamran Asif

Abstract:

MPLS stands for Multi-Protocol Label Switching. It is the technology which replaces ATM (Asynchronous Transfer Mode) and frame relay. In this paper, we have designed a full fledge small scale MPLS based service provider network core network model, which provides communication services (e.g. voice, video and data) to the customer more efficiently using label switching technique. Using MPLS VPN provides security to the customers which are either on LAN or WAN. It protects its single customer sites from being attacked by any intruder from outside world along with the provision of concept of extension of a private network over an internet. In this paper, we tried to implement a service provider network using minimum available resources i.e. five 3800 series CISCO routers comprises of service provider core, provider edge routers and customer edge routers. The customers on the one end of the network (customer side) is capable of sending any kind of data to the customers at the other end using service provider cloud which is MPLS VPN enabled. We have also done simulation and emulation for the model using GNS3 (Graphical Network Simulator-3) and achieved the real time scenarios. We have also deployed a NMS system which monitors our service provider cloud and generates alarm in case of any intrusion or malfunctioning in the network. Moreover, we have also provided a video help desk facility between customers and service provider cloud to resolve the network issues more effectively.

Keywords: MPLS, VPN, NMS, ATM, asynchronous transfer mode

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3187 Microfacies Analysis and Paleoenvironmental Trends of the Paleocene Farrud and Mabruk Reservoirs, Concession 11, West Sirte Basin, Libya

Authors: Nisreen Agha

Abstract:

Investigation of representative core samples under the petrological microscope reveals common petrographic and mineralogical characteristics with distinct faunal assemblages, allowing establishing of the microfacies associations and deducing the paleoenvironmental trends of the Paleocene Farrud and Mabruk rock units. Recognition of the early and post-diagenetic processes, particularly dolomitization and micritization, as well as dissolution and precipitation of spary drusy calcite as a new morphism process affecting the reservoir rocks, is established. The microfacies trends detected from the investigation of 46 core samples from Farrud member (Lower Paleocene) representing six wells; QQQ1-11, GG1-11, LLL1-11, RRR1-11, RRR40-11, and RRR45-11 indicate that the deposition was started within the realm of shallow supratidal and intertidal subenvironments followed by deeper environments of the shelf bays with maximum sea level during inner shelf environment where fossiliferous bioclastic packstone dominated. The microfacies associations determined in 8 core samples from two wells LLL1and RRR40 representing Mabruk Member (Upper Paleocene), indicate paleoenvironmental trends marked by sea level fluctuations accompanied with a relatively marine shelf bay conditions intervened with short-lived shallow intertidal and supratidal warm coastal sedimentation. As a result, dolostone, evaporitic dismicrites, and gypsiferous dolostone of supratidal characters were deposited. They reflect rapid oscillation of the sea level marked by drop land-ward shift of the sea shore deposition prevailed by supratidal gypsiferous dolostone and numerous ferruginous materials as clouds straining many parts of dolomite and surrounded the micritized fossils. This situation ends the deposition of the Farrud Member in most of the studied wells. On the other hand, the facies in the northern part of the Concession -11 field indicates deposition in a deeper marine setting than in the southern facies.

Keywords: Farrud and Mabruk members, paleocene, microfacies associations, diagenesis, sea level oscillation, depositional environments

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3186 Safety Assessment and Prophylactic Efficacy of Moringa stenopetala Leaf Extract Through Mitigation of Oxidative Stress in BV-2 Microglial Cell

Authors: Stephen Adeniyi Adefegha, Vitor Mostardeiro, Vera Maria Morsch, Ademir F. Morel, Ivana Beatrice Manica Da Cruz, Sabrina Somacal Maria Rosa Chitolina Schetinger

Abstract:

Moringa stenopetala is often consumed as food and used in folkloric medicine for the management of several diseases. Purpose: This study was set up in order to assess the effect of aqueous extract of Moringa stenopetala on cell viability and oxidative stress biomarkers in BV-2 microglial cells. Aqueous extracts of M. stenopetala were prepared, lyophilized and reconstituted in 0.5% dimethylsulphoxide (DMSO). Cells were treated with M. stenopetala extracts (0.1 - 100 µg/ml) for cell viability and nitric oxide (NO) production tests. However, M. stenopetala extract (50 µg/ml) was used in the treatment of cells for the determination of protein carbonyl content and reactive oxygen species (ROS) level. Incubation of BV-2 microglia cell with M. stenopetala extract maintained cell viability, diminished NO and ROS levels, and reduced protein carbonyl contents Chlorogenic acid, rutin, kaempferol and quercetin derivatives were the main phenolic compounds identified in M. stenopetala leaf extract. These phenolic compounds present in M. stenopetala may be responsible for the mitigation of oxidative stress in BV-2 microglial cells.

Keywords: oxidative stress, BV-2 microglial cell, Moringa stenopetala, cell viability, antioxidant

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3185 Prevalence and Associated Factors of Protein-Energy Malnutrition Among Children Aged 6-59 Months in Babile Town from April to June 2016

Authors: Tajudin Ahmed

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Malnutrition is a significant problem in developing countries, particularly among children, due to inadequate diets, lack of proper care, and unequal distribution of food within households. High rates of malnutrition have been shown in Ethiopia, including stunting, underweight, and wasting. This study aims to assess the prevalence and associated factors of Protein-Energy Malnutrition (PEM) among children aged 6-59 months in Babile Town. The study utilized a community-based cross-sectional design conducted in Babile Town, Eastern Ethiopia. Two kebeles were randomly selected, and a census was conducted to identify eligible households. A total of 391 households with children aged 6-59 months were included in the study. Data was collected using structured questionnaires, and anthropometric measurements were taken to assess the weight and height of the children. The study found that a majority of the mothers (72.34%) and fathers (43%) had no formal education. Among the mothers who could read and write, a small percentage had completed primary (14%) or secondary (14%) education, and even fewer had higher education (2.7%). Similarly, among the fathers who could read and write, a majority had completed primary (46.15%) or secondary (27.22%) education, with smaller percentages completing preparatory (8.4%) or higher education (6.29%). The prevalence of malnutrition in the study area was high, with 38.85% of children experiencing stunting (8.2% severely stunted), 50.13% wasting (9% severely wasted), and 41.43% underweight (6.65% severely underweight). These findings indicate a significant burden of malnutrition in Babile Town, likely exacerbated by the high prevalence of infectious diseases such as diarrhea. The study concludes that the prevalence of malnutrition, particularly stunting, wasting, and underweight, is high in Babile Town. The findings indicate the urgent need for interventions to address malnutrition and improve nutrition and healthcare practices in the study area. These results can serve as a baseline for future studies and inform policymakers and healthcare providers in their efforts to combat childhood malnutrition.

Keywords: protein-energy malnutrition, children 6-59 month age babble town, Marasmus

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3184 Loss of Function of Only One of Two CPR5 Paralogs Causes Resistance Against Rice Yellow Mottle Virus

Authors: Yugander Arra, Florence Auguy, Melissa Stiebner, Sophie Chéron, Michael M. Wudick, Van Schepler-Luu, Sébastien Cunnac, Wolf B. Frommer, Laurence Albar

Abstract:

Rice yellow mottle virus (RYMV) is one of the most important diseases affecting rice in Africa. The most promising strategy to reduce yield losses is the use of highly resistant varieties. The resistance gene RYMV2 is homolog of the Arabidopsis constitutive expression of pathogenesis related protein-5 (AtCPR5) nucleoporin gene. Resistance alleles are originating from African cultivated rice Oryza glaberrima, rarely cultivated, and are characterized by frameshifts or early stop codons, leading to a non-functional or truncated protein. Rice possesses two paralogs of CPR5 and function of these genes are unclear. Here, we evaluated the role of the two rice candidate nucleoporin paralogs OsCPR5.1 (pathogenesis-related gene 5; RYMV2) and OsCPR5.2 by CRISPR/Cas9 genome editing. Despite striking sequence and structural similarity, only loss-of-function of OsCPR5.1 led to full resistance, while loss-of-function oscpr5.2 mutants remained susceptible. Short N-terminal deletions in OsCPR5.1 also did not lead to resistance. In contrast to Atcpr5 mutants, neither OsCPR5.1 nor OsCPR5.2 knock out mutants showed substantial growth defects. Taken together, the candidate nucleoporin OsCPR5.1, but not its close homolog OsCPR5.2, plays a specific role for the susceptibility to RYMV, possibly by impairing the import of viral RNA or protein into the nucleus. Whereas gene introgression from O. glaberrima to high yielding O. sativa varieties is impaired by strong sterility barriers and the negative impact of linkage drag, genome editing of OsCPR5.1, while maintaining OsCPR5.2 activity, thus provides a promising strategy to generate O. sativa elite lines that are resistant to RYMV.

Keywords: CRISPR Cas9, genome editing, knock out mutant, recessive resistance, rice yellow mottle virus

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3183 Effect of Mistranslating tRNA Alanine on Polyglutamine Aggregation

Authors: Sunidhi Syal, Rasangi Tennakoon, Patrick O'Donoghue

Abstract:

Polyglutamine (polyQ) diseases are a group of diseases related to neurodegeneration caused by repeats of the amino acid glutamine (Q) in the DNA, which translates into an elongated polyQ tract in the protein. The pathological explanation is that the polyQ tract forms cytotoxic aggregates in the neurons, leading to their degeneration. There are no cures or preventative efforts established for these diseases as of today, although the symptoms of these diseases can be relieved. This study specifically focuses on Huntington's disease, which is a type of polyQ disease in which aggregation is caused by the extended cytosine, adenine, guanine (CUG) codon repeats in the huntingtin (HTT) gene, which encodes for the huntingtin protein. Using this principle, we attempted to create six models, which included mutating wildtype tRNA alanine variant tRNA-AGC-8-1 to have glutamine anticodons CUG and UUG so serine is incorporated at glutamine sites in poly Q tracts. In the process, we were successful in obtaining tAla-8-1 CUG mutant clones in the HTTexon1 plasmids with a polyQ tract of 23Q (non-pathogenic model) and 74Q (disease model). These plasmids were transfected into mouse neuroblastoma cells to characterize protein synthesis and aggregation in normal and mistranslating cells and to investigate the effects of glutamines replaced with alanines on the disease phenotype. Notably, we observed no noteworthy differences in mean fluorescence between the CUG mutants for 23Q or 74Q; however, the Triton X-100 assay revealed a significant reduction in insoluble 74Q aggregates. We were unable to create a tAla-8-1 UUG mutant clone, and determining the difference in the effects of the two glutamine anticodons may enrich our understanding of the disease phenotype. In conclusion, by generating structural disruption with the amino acid alanine, it may be possible to find ways to minimize the toxicity of Huntington's disease caused by these polyQ aggregates. Further research is needed to advance knowledge in this field by identifying the cellular and biochemical impact of specific tRNA variants found naturally in human genomes.

Keywords: Huntington's disease, polyQ, tRNA, anticodon, clone, overlap PCR

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