Search results for: chaperone assisted protein folding
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3174

Search results for: chaperone assisted protein folding

2514 Nanoparticles-Protein Hybrid-Based Magnetic Liposome

Authors: Amlan Kumar Das, Avinash Marwal, Vikram Pareek

Abstract:

Liposome plays an important role in medical and pharmaceutical science as e.g. nano scale drug carriers. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment. Magnet-driven liposome used for the targeted delivery of drugs to organs and tissues1. These liposome preparations contain encapsulated drug components and finely dispersed magnetic particles. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment that are generated in vitro. These are useful in terms of biocompatibility, biodegradability, and low toxicity, and can control biodistribution by changing the size, lipid composition, and physical characteristics2. Furthermore, liposomes can entrap both hydrophobic and hydrophilic drugs and are able to continuously release the entrapped substrate, thus being useful drug carriers. Magnetic liposomes (MLs) are phospholipid vesicles that encapsulate magneticor paramagnetic nanoparticles. They are applied as contrast agents for magnetic resonance imaging (MRI)3. The biological synthesis of nanoparticles using plant extracts plays an important role in the field of nanotechnology4. Green-synthesized magnetite nanoparticles-protein hybrid has been produced by treating Iron (III)/Iron(II) chloride with the leaf extract of Dhatura Inoxia. The phytochemicals present in the leaf extracts act as a reducing as well stabilizing agents preventing agglomeration, which include flavonoids, phenolic compounds, cardiac glycosides, proteins and sugars. The magnetite nanoparticles-protein hybrid has been trapped inside the aqueous core of the liposome prepared by reversed phase evaporation (REV) method using oleic and linoleic acid which has been shown to be driven under magnetic field confirming the formation magnetic liposome (ML). Chemical characterization of stealth magnetic liposome has been performed by breaking the liposome and release of magnetic nanoparticles. The presence iron has been confirmed by colour complex formation with KSCN and UV-Vis study using spectrophotometer Cary 60, Agilent. This magnet driven liposome using nanoparticles-protein hybrid can be a smart vesicles for the targeted drug delivery.

Keywords: nanoparticles-protein hybrid, magnetic liposome, medical, pharmaceutical science

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2513 Radial Basis Surrogate Model Integrated to Evolutionary Algorithm for Solving Computation Intensive Black-Box Problems

Authors: Abdulbaset Saad, Adel Younis, Zuomin Dong

Abstract:

For design optimization with high-dimensional expensive problems, an effective and efficient optimization methodology is desired. This work proposes a series of modification to the Differential Evolution (DE) algorithm for solving computation Intensive Black-Box Problems. The proposed methodology is called Radial Basis Meta-Model Algorithm Assisted Differential Evolutionary (RBF-DE), which is a global optimization algorithm based on the meta-modeling techniques. A meta-modeling assisted DE is proposed to solve computationally expensive optimization problems. The Radial Basis Function (RBF) model is used as a surrogate model to approximate the expensive objective function, while DE employs a mechanism to dynamically select the best performing combination of parameters such as differential rate, cross over probability, and population size. The proposed algorithm is tested on benchmark functions and real life practical applications and problems. The test results demonstrate that the proposed algorithm is promising and performs well compared to other optimization algorithms. The proposed algorithm is capable of converging to acceptable and good solutions in terms of accuracy, number of evaluations, and time needed to converge.

Keywords: differential evolution, engineering design, expensive computations, meta-modeling, radial basis function, optimization

Procedia PDF Downloads 387
2512 Development and Investigation of Efficient Substrate Feeding and Dissolved Oxygen Control Algorithms for Scale-Up of Recombinant E. coli Cultivation Process

Authors: Vytautas Galvanauskas, Rimvydas Simutis, Donatas Levisauskas, Vykantas Grincas, Renaldas Urniezius

Abstract:

The paper deals with model-based development and implementation of efficient control strategies for recombinant protein synthesis in fed-batch E.coli cultivation processes. Based on experimental data, a kinetic dynamic model for cultivation process was developed. This model was used to determine substrate feeding strategies during the cultivation. The proposed feeding strategy consists of two phases – biomass growth phase and recombinant protein production phase. In the first process phase, substrate-limited process is recommended when the specific growth rate of biomass is about 90-95% of its maximum value. This ensures reduction of glucose concentration in the medium, improves process repeatability, reduces the development of secondary metabolites and other unwanted by-products. The substrate limitation can be enhanced to satisfy restriction on maximum oxygen transfer rate in the bioreactor and to guarantee necessary dissolved carbon dioxide concentration in culture media. In the recombinant protein production phase, the level of substrate limitation and specific growth rate are selected within the range to enable optimal target protein synthesis rate. To account for complex process dynamics, to efficiently exploit the oxygen transfer capability of the bioreactor, and to maintain the required dissolved oxygen concentration, adaptive control algorithms for dissolved oxygen control have been proposed. The developed model-based control strategies are useful in scale-up of cultivation processes and accelerate implementation of innovative biotechnological processes for industrial applications.

Keywords: adaptive algorithms, model-based control, recombinant E. coli, scale-up of bioprocesses

Procedia PDF Downloads 250
2511 In Silico Analysis of Deleterious nsSNPs (Missense) of Dihydrolipoamide Branched-Chain Transacylase E2 Gene Associated with Maple Syrup Urine Disease Type II

Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed

Abstract:

Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.

Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer

Procedia PDF Downloads 197
2510 Proteomic Analysis of Cytoplasmic Antigen from Brucella canis to Characterize Immunogenic Proteins Responded with Naturally Infected Dogs

Authors: J. J. Lee, S. R. Sung, E. J. Yum, S. C. Kim, B. H. Hyun, M. Her, H. S. Lee

Abstract:

Canine brucellosis is a critical problem in dogs leading to reproductive diseases which are mainly caused by Brucella canis. There are, nonetheless, not clear symptoms so that it may go unnoticed in most of the cases. Serodiagnosis for canine brucellosis has not been confirmed. Moreover, it has substantial difficulties due to broad cross-reactivity between the rough cell wall antigens of B. canis and heterospecific antibodies present in normal, uninfected dogs. Thus, this study was conducted to characterize the immunogenic proteins in cytoplasmic antigen (CPAg) of B. canis, which defined the antigenic sensitivity of the humoral antibody responses to B. canis-infected dogs. In analysis of B. canis CPAg, first, we extracted and purified the cytoplasmic proteins from cultured B. canis by hot-saline inactivation, ultrafiltration, sonication, and ultracentrifugation step by step according to the sonicated antigen extract method. For characterization of this antigen, we checked the sort and range of each protein on SDS-PAGE and verified the immunogenic proteins leading to reaction with antisera of B. canis-infected dogs. Selected immunodominant proteins were identified using MALDI-MS/MS. As a result, in an immunoproteomic assay, several polypeptides in CPAg on one or two-dimensional electrophoresis (DE) were specifically reacted to antisera from B. canis-infected dogs but not from non-infected dogs. The polypeptides with approximate 150, 80, 60, 52, 33, 26, 17, 15, 13, 11 kDa on 1-DE were dominantly recognized by antisera from B. canis-infected dogs. In the immunoblot profiles on 2-DE, ten immunodominant proteins in CPAg were detected with antisera of infected dogs between pI 3.5-6.5 at approximate 35 to 10 KDa, without any nonspecific reaction with sera in non-infected dogs. Ten immunodominant proteins identified by MALDI-MS/MS were identified as superoxide dismutase, bacteroferritin, amino acid ABC transporter substrate-binding protein, extracellular solute-binding protein family3, transaldolase, 26kDa periplasmic immunogenic protein, Rhizopine-binding protein, enoyl-CoA hydratase, arginase and type1 glyceraldehyde-3-phosphate dehydrogenase. Most of these proteins were determined by their cytoplasmic or periplasmic localization with metabolism and transporter functions. Consequently, this study discovered and identified the prominent immunogenic proteins in B. canis CPAg, highlighting that those antigenic proteins may accomplish a specific serodiagnosis for canine brucellosis. Furthermore, we will evaluate those immunodominant proteins for applying to the advanced diagnostic methods with high specificity and accuracy.

Keywords: Brucella canis, Canine brucellosis, cytoplasmic antigen, immunogenic proteins

Procedia PDF Downloads 137
2509 Chemical Composition Pistachio Harvested Bechloul (Algeria)

Authors: Nadjiba Mezıou-Cheboutı, Amel Merabet, Yahia Cheboutı, Nassima Behidj, Fatima-Zohra Bıssaad et Salahedine Doumandjı

Abstract:

Among the Anacardiaceae, the fruit (Pistacia vera L.) is the only species that produces edible fruits. The introduction of real pistachio was made in the early sixties by an FAO program in Algeria in several regions in the northern part of Algeria: Tlemcen, Sidi Bel Abbes, Batna, Bouira M'sila . Chemical analyzes of seeds pistachios were made on seeds from an orchard that localizes to Bechloul (Bouira) located in bioclimatic sub-humid temperate winter floor. Analyzes reveal dry matter content of 3.60 ± 0.45%, the water rate is 7.21 ± 0.36%. However, the fat content is 46.00 ± 0.90%, in average blood sugar, it is 4.02 ± 0.47%, the protein reached 29.88 ± 0.76%. Given the very interesting that high-fat food nutritional values, culture pistachio must be considered for its extension in Algeria.

Keywords: pistachio, dry matter, fat, sugar, protein

Procedia PDF Downloads 344
2508 Microencapsulation of Tuna Oil and Mentha Piperita Oil Mixture using Different Combinations of Wall Materials with Whey Protein Isolate

Authors: Amr Mohamed Bakry Ibrahim, Yingzhou Ni, Hao Cheng, Li Liang

Abstract:

Tuna oil (omega-3 oil) has become increasingly popular in the last ten years, because it is considered one of the treasures of food which has many beneficial health effects for the humans. Nevertheless, the susceptibility of omega-3 oils to oxidative deterioration, resulting in the formation of oxidation products, in addition to organoleptic problems including “fishy” flavors, have presented obstacles to the more widespread use of tuna oils in the food industry. This study sought to evaluate the potential impact of Mentha piperita oil on physicochemical characteristics and oxidative stability of tuna oil microcapsules formed by spray drying using the partial substitution to whey protein isolate by carboxymethyl cellulose and pullulan. The emulsions before the drying process were characterized regarding size and ζ-potential, viscosity, surface tension. Confocal laser scanning microscopy showed that all emulsions were sphericity and homogeneous distribution without any visible particle aggregation. The microcapsules obtained after spray drying were characterized regarding microencapsulation efficiency, water activity, color, bulk density, flowability, scanning surface morphology and oxidative stability. The microcapsules were spherical shape had low water activity (0.11-0.23 aw). The microcapsules containing both tuna oil and Mentha piperita oil were smaller than others and addition of pullulan into wall materials improved the morphology of microcapsules. Microencapsulation efficiency of powdered oil ranged from 90% to 94%. Using Mentha piperita oil in the process of microencapsulation tuna oil enhanced the oxidative stability using whey protein isolate only or with carboxymethyl cellulose or pullulan as wall materials, resulting in improved storage stability and mask fishy odor. Therefore, it is foreseen using tuna-Mentha piperita oil mixture microcapsules in the applications of the food industries.

Keywords: Mentha piperita oil, microcapsule, tuna oil, whey protein isolate

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2507 Influence of Environmental Conditions on a Solar Assisted Mashing Process

Authors: Ana Fonseca, Stefany Villacis

Abstract:

In this paper, the influence of several scenarios on a model of solar assisted mashing process in a brewery, while applying the model to different locations and therefore changing the environmental conditions, was analyzed. Assorted beer producer locations in different countries around the globe with contrasting climatic zones such as Guayaquil (Ecuador), Bangkok (Thailand), Mumbai (India), Veracruz (Mexico) and Brisbane (Australia) were evaluated and compared with a base case study Oldenburg (Germany), and results were drawn. The evaluation was restricted to the results obtained using TRNSYS 16 as simulating tool. On the base case, an annual Solar Fraction (SF) of 0.50 was encountered, results showed highly affection when modifying the pump control of the primary circuit and when increasing the area of collectors. A sensitivity analysis of the system for the selected locations was performed, resulting in Guayaquil the highest annual SF with a ratio of 2.5 times the expected value as compared with the base case. In contrast, Brisbane presented the lowest ratio, resulting in half of the expected one due to its lower irradiance. In conclusion, cities in Sunbelt countries have the technical potential to apply solar heat for their low-temperature industrial processes, in this case implementing a green brewery in Guayaquil.

Keywords: evacuated tubular solar collector, irradiance, mashing process, solar fraction, solar thermal

Procedia PDF Downloads 128
2506 Development and Compositional Analysis of Functional Bread and Biscuit from Soybean, Peas and Rice Flour

Authors: Jean Paul Hategekimana, Bampire Claudine, Niyonsenga Nadia, Irakoze Josiane

Abstract:

Peas, soybeans and rice are crops which are grown in Rwanda and are available in rural and urban local markets and they give contribution in reduction of health problems especially in fighting malnutrition and food insecurity in Rwanda. Several research activities have been conducted on how cereals flour can be mixed with legumes flour for developing baked products which are rich in protein, fiber, minerals as they are found in legumes. However, such activity was not yet well studied in Rwanda. The aim of the present study was to develop bread and biscuit products from peas, soybeans and rice as functional ingredients combined with wheat flour and then analyze the nutritional content and consumer acceptability of new developed products. The malnutrition problem can be reduced by producing bread and biscuits which are rich in protein and are very accessible for every individual. The processing of bread and biscuit were made by taking peas flour, soybeans flour and rice flour mixed with wheat flour and other ingredients then a dough was made followed by baking. For bread, two kind of products were processed, for each product one control and three experimental samples in different three ratios of peas and rice were prepared. These ratios were 95:5, 90:10 and 80:20 for bread from peas and 85:5:10, 80:10:10 and 70:10:20 for bread from peas and rice. For biscuit, two kind of products were also processed, for each product one control sample and three experimental samples in three different ratios were prepared. These ratios are 90:5:5,80:10:10 and 70:10:20 for biscuit from peas and rice and 90:5:5,80:10:10 and 70:10:20 for biscuit from soybean and rice. All samples including the control sample were analyzed for the consumer acceptability (sensory attributes) and nutritional composition. For sensory analysis, bread from of peas and rice flour with wheat flour at ratio 85:5:10 and bread from peas only as functional ingredient with wheat flour at ratio 95:5 and biscuits made from a of soybeans and rice at a ratio 90:5:5 and biscuit made from peas and rice at ratio 90:5:5 were most acceptable compared to control sample and other samples in different ratio. The moisture, protein, fat, fiber and minerals (Sodium and iron.) content were analyzed where bread from peas in all ratios was found to be rich in protein and fiber compare to control sample and biscuit from soybean and rice in all ratios was found to be rich in protein and fiber compare to control sample.

Keywords: bakery products, peas and rice flour, wheat flour, sensory evaluation, proximate composition

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2505 Optimization of Ultrasound Assisted Extraction and Characterization of Functional Properties of Dietary Fiber from Oat Cultivar S2000

Authors: Muhammad Suhail Ibrahim, Muhammad Nadeem, Waseem Khalid, Ammara Ainee, Taleeha Roheen, Sadaf Javaria, Aftab Ahmed, Hira Fatima, Mian Nadeem Riaz, Muhammad Zubair Khalid, Isam A. Mohamed Ahmed J, Moneera O. Aljobair

Abstract:

This study was executed to explore the efficacy of ultrasound-assisted extraction of dietary fiber from oat cultivar S2000. Extraction (variables time, temperature and amplitude) was optimized by using response surface methodology (RSM) conducted by Box Behnken Design (BBD). The effect of time, temperature and amplitude were studied at three levels. It was observed that time and temperature exerted more impact on extraction efficiency as compared to amplitude. The highest yield of total dietary fiber (TDF), soluble dietary fiber (SDF) and In-soluble dietary fiber (IDF) fractions were observed under ultrasound processing for 20 min at 40 ◦C with 80% amplitude. Characterization of extracted dietary fiber showed that it had better crystallinity, thermal properties and good fibrous structure. It also showed better functional properties as compared to traditionally extracted dietary fiber. Furthermore, dietary fibers from oats may offer high-value utilization and the expansion of comprehensive utilization in functional food and nutraceutical development.

Keywords: extraction, ultrasonication, response surface methodology, box behnken design

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2504 Structural and Magnetic Properties of NiFe2O4 Spinel Ferrite Nanoparticles Synthesized by Starch-Assisted Sol-Gel Auto-Combustion Method

Authors: R. S. Yadav, J. Havlica, I. Kuřitka, Z. Kozakova, J. Masilko, L. Kalina, M. Hajdúchová, V. Enev, J. Wasserbauer

Abstract:

Nickel spinel ferrite NiFe2O4 nanoparticles with different particle size at different annealing temperature were synthesized using the starch-assisted sol-gel auto-combustion method. The synthesized nanoparticles were characterized by conventional powder X-ray diffraction (XRD) spectroscopy, Raman Spectroscopy, Fourier Transform Infrared Spectroscopy, Field-Emission Scanning Electron Microscopy, X-ray Photoelectron Spectroscopy and Vibrating Sample Magnetometer. The XRD patterns confirmed the formation of NiFe2O4 spinel ferrite nanoparticles. Field-Emission Scanning Electron Microscopy revealed that particles are of spherical morphology with particle size 5-20 nm at lower annealing temperature. An infrared spectroscopy study showed the presence of two principal absorption bands in the frequency range around 525 cm-1 (ν1) and around 340 cm-1 (ν2); which indicate the presence of tetrahedral and octahedral group complexes, respectively, within the spinel ferrite nanoparticles. Raman spectroscopy study also indicated the change in octahedral and tetrahedral site related Raman modes in nickel ferrite nanoparticles with change of particle size. This change in magnetic behavior with change of particle size of NiFe2O4 nanoparticles was observed.

Keywords: nickel ferrite, nanoparticles, magnetic property, NiFe2O4

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2503 The Chemical Composition of the Pistachio (Pistacia vera) Harvested Bechloul (Algeria)

Authors: Nadjiba Meziou-Chebouti, Amel Merabet, Yahia Chebouti, Nassima Behidj, Salahedine Doumandji

Abstract:

Among the Anacardiaceae, the fruit (Pistacia vera L.) is the only species that produces edible fruits. The introduction of real pistachio was made in the early sixties by an FAO program in Algeria in several regions in the northern part of Algeria: Tlemcen, Sidi Bel Abbes, Batna, Bouira M'sila. Chemical analyzes of seeds pistachios were made on seeds from an orchard that localizes to Bechloul (Bouira) located in bioclimatic sub-humid temperate winter floor. Analyzes reveal dry matter content of 3.60±0.45%, the water rate is 7.21±0.36%. However, the fat content is 46.00±0.90%, in average blood sugar, it is 4.02±0.47%, the protein reached 29.88±0.76%. Given the very interesting that high-fat food nutritional values, culture pistachio must be considered for its extension in Algeria.

Keywords: pistachio, dry matter, fat, sugar, protein

Procedia PDF Downloads 356
2502 Using Audio-Visual Aids and Computer-Assisted Language Instruction to Overcome Learning Difficulties of Vocabulary in Students of Special Needs

Authors: Sadeq Al Yaari, Ayman Al Yaari, Adham Al Yaari, Montaha Al Yaari, Aayah Al Yaari, Sajedah Al Yaar

Abstract:

Objectives: To assess the effect of using audio-visual aids and computer-assisted/ aided language instruction (CALI) in the performance of students of special needs studying vocabulary course. Methods: The performance of forty students of special needs (males and females) who used audiovisual aids and CALI in their vocabulary course at al-Malādh school for students of special needs was compared to that of another group (control group) of the same number and age (8-18). Again, subjects in the experimental group were given lessons using audio-visual aids and CALI, while those in the control group were given lessons using ordinary educational aids only, although both groups almost shared the same features (class environment, speech language therapist (SLT), etc.). Pre-andposttest was given at the beginning and end of the semester and a qualitative and quantitative analysis followed. Results & conclusions: Results of the present experimental study's pre-and-posttests indicated that the performance of the students in the first group was higher than that of those of the second group (34.27%, 73.82% vs. 33.57%, 34.92%, respectively). Compared with females, males’ performance was higher (1515 scores vs. 1438 scores). Such findings suggest that the presence of these audiovisual aids and CALI in the classes of students of special needs, especially if they are studying vocabulary building course is very important due to their usefulness in the improvement of performance of the students of special needs.

Keywords: language components, vocabulary, audio-visual aids, CALI, special needs, students, SLTs

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2501 Development of an Advanced Power Ultrasonic-Assisted Drilling System

Authors: M. A. Moghaddas, M. Short, N. Wiley, A. Y. Yi, K. F. Graff

Abstract:

The application of ultrasonic vibrations to machining processes has a long history, ranging from slurry-based systems able to drill brittle materials, to more recent developments involving low power ultrasonics for high precision machining, with many of these at the research and laboratory stages. The focus of this development is the application of high levels of ultrasonic power (1,000’s of watts) to standard, heavy duty machine tools – drilling being the immediate focus, with developments in milling in progress – with the objective of dramatically increasing system productivity through faster feed rates, this benefit arising from the thrust force reductions obtained by power ultrasonic vibrations. The presentation will describe development of an advanced drilling system based on a special, acoustically designed, rugged drill module capable of functioning under heavy duty production conditions, and making use of standard tool holder means, and able to obtain thrust force reductions while maintaining or improving surface finish and drilling accuracy. The characterization of the system performance will be described, and results obtained in drilling several materials (Aluminum, Stainless steel, Titanium) presented.

Keywords: dimensional accuracy, machine tool, productivity, surface roughness, thrust force, ultrasonic vibrations, ultrasonic-assisted drilling

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2500 Influence of Physico-Chemical Changes in the Environment on the Behavior of Tadpoles Rana Saharica: Case of Fungicide (Artea 330ec)

Authors: S. Zouainia, R. Djebar, H. Berrebah, A. Sayeb

Abstract:

This work focused on the study of physiological and biochemical changes observed in tadpoles exposed to fungicide Rana saharica Artea 330ec recently introduced in Algeria. For this, we tested the effect of xenobiotics on growth and development of tadpoles; among the studied parameters: total protein, glutathione and respiratory activity. The study of physiological parameters showed that the tadpoles change perfectly in the absence of toxic and in favorable conditions (pH, temperature). Our results showed an increased rate of protein and GSH in the presence of the fungicide Artea 330ec. The latter causes uninhibited very highly significant respiratory activity of tadpoles treated. The presence of xenobiotics in the breeding tadpoles water causes disturbances in behavior and food metabolism.

Keywords: amphibians, fungicides, pesticides, pollution

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2499 Phage Capsid for Efficient Delivery of Cytotoxic Drugs

Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

Abstract:

The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

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2498 Influence of JHA and Ecdysteroid on Reproduction in Dysdercus similis (Hemiptera: Pyrrhocoridae)

Authors: Versha Sharma

Abstract:

Juvenile hormone analogue, fenoxycarb and ecdysterone, when applied at varying concentrations in the adult females of Dysdercus similis, in situ histochemical observations of treated ovarian and adipose tissues during the first gonotrophic cycle elicited drastic histomorphological changes in both tissues. The action and effect of both JHa and ecdysterone on ovarian development, vitellogenesis, the activity of follicular epithelium, chorion formation all were monitored in detail. SDS-PAGE electrophoretic analysis showed drastic downregulation on the protein profile of differently treated tissue samples. After exogenous JHa supply, resorption of the developing oocytes was also often noticed. Gradational decline and disappearance of different protein bands in treated both ovarian and adipose tissues noticed could be due to the depletion of specific metabolites essential for oocyte development and maturation. Natural products support both crop production and the environment that being effective in pest control, less toxic to non-target organisms and at the same time biodegradable. Hence, these could be utilized as an attractive alternative to the synthetic chemical insecticides for at least cotton bug pest management. Increasing IGR dosages is found to elicit both qualitative and quantitative depletion of protein metabolites and drastic histochemical changes in the gonads of the treated forms brought forth the production of a large number of immature mal-formed oocytes. Findings in greater detail could be discussed.

Keywords: juvenile hormone, ecdysone, P. picta, Dysdercus similis

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2497 Protective Effect of Bexarotene, a Selective RXRα Agonist, against Hypotension Associated with Inflammation and Tissue Injury Linked to Decreased Circulating iNOS Levels in A Rat Model of Septic Shock

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat

Abstract:

We hypothesized that rexinoids such as bexarotene, a selective retinoid X receptor α (RXRα) agonist, may be beneficial for preventing mortality due to inflammation associated with increased expression/activity of inducible nitric oxide synthase (iNOS) induced by lipopolysaccharide (LPS). Therefore, we investigated effects of bexarotene on the changes in circulating protein levels of iNOS (an index for systemic iNOS expression), myeloperoxidase (MPO) (an index for systemic inflammation), and lactate dehydrogenase (LDH) (an index for systemic tissue injury) in LPS-induced systemic inflammation model resulting in septic shock in rats. Rats were injected with saline (4 ml/kg; i.p.), LPS (10 mg/kg; i.p.), dimethylsulphoxide (4 ml/kg, 0.1%; s.c.) at time 0. Mean arterial blood pressure and heart rate were measured using a tail-cuff device. Bexarotene (0.03, 0.1, 0.3, and 1 mg/kg; s.c.) was administered to separate groups of rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and blood was collected for measurement of serum iNOS, MPO, and LDH protein levels. Blood pressure decreased by 31 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. Bexarotene at 0.3 and 1 mg/kg doses caused 20% mortality 4 h after LPS injection. In the LPS-treated rats, serum iNOS, MPO, and LDH protein levels were increased. Bexarotene only at 0.1 mg/kg dose prevented the LPS-induced hypotension and increased in iNOS, MPO, and LDH protein levels. These data are consistent with the view that a decrease in systemic iNOS levels contributes to the beneficial effect of bexarotene to prevent the hypotension associated with inflammation and tissue injury during rat endotoxemia. [This work was financially supported by The Scientific and Technological Research Council of Turkey (SBAG-109S121)].

Keywords: bexarotene, inflammation, iNOS, lipopolisaccharide, RXRa

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2496 Mesoporous Titania Thin Films for Gentamicin Delivery and Bone Morphogenetic Protein-2 Immobilization

Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

Abstract:

The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

Procedia PDF Downloads 158
2495 Evaluation of Functional Properties of Protein Hydrolysate from the Fresh Water Mussel Lamellidens marginalis for Nutraceutical Therapy

Authors: Jana Chakrabarti, Madhushrita Das, Ankhi Haldar, Roshni Chatterjee, Tanmoy Dey, Pubali Dhar

Abstract:

High incidences of Protein Energy Malnutrition as a consequence of low protein intake are quite prevalent among the children in developing countries. Thus prevention of under-nutrition has emerged as a critical challenge to India’s developmental Planners in recent times. Increase in population over the last decade has led to greater pressure on the existing animal protein sources. But these resources are currently declining due to persistent drought, diseases, natural disasters, high-cost of feed, and low productivity of local breeds and this decline in productivity is most evident in some developing countries. So the need of the hour is to search for efficient utilization of unconventional low-cost animal protein resources. Molluscs, as a group is regarded as under-exploited source of health-benefit molecules. Bivalve is the second largest class of phylum Mollusca. Annual harvests of bivalves for human consumption represent about 5% by weight of the total world harvest of aquatic resources. The freshwater mussel Lamellidens marginalis is widely distributed in ponds and large bodies of perennial waters in the Indian sub-continent and well accepted as food all over India. Moreover, ethno-medicinal uses of the flesh of Lamellidens among the rural people to treat hypertension have been documented. Present investigation thus attempts to evaluate the potential of Lamellidens marginalis as functional food. Mussels were collected from freshwater ponds and brought to the laboratory two days before experimentation for acclimatization in laboratory conditions. Shells were removed and fleshes were preserved at- 20oC until analysis. Tissue homogenate was prepared for proximate studies. Fatty acids and amino acids composition were analyzed. Vitamins, Minerals and Heavy metal contents were also studied. Mussel Protein hydrolysate was prepared using Alcalase 2.4 L and degree of hydrolysis was evaluated to analyze its Functional properties. Ferric Reducing Antioxidant Power (FRAP) and DPPH Antioxidant assays were performed. Anti-hypertensive property was evaluated by measuring Angiotensin Converting Enzyme (ACE) inhibition assay. Proximate analysis indicates that mussel meat contains moderate amount of protein (8.30±0.67%), carbohydrate (8.01±0.38%) and reducing sugar (4.75±0.07%), but less amount of fat (1.02±0.20%). Moisture content is quite high but ash content is very low. Phospholipid content is significantly high (19.43 %). Lipid constitutes, substantial amount of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) which have proven prophylactic values. Trace elements are found present in substantial amount. Comparative study of proximate nutrients between Labeo rohita, Lamellidens and cow’s milk indicates that mussel meat can be used as complementary food source. Functionality analyses of protein hydrolysate show increase in Fat absorption, Emulsification, Foaming capacity and Protein solubility. Progressive anti-oxidant and anti-hypertensive properties have also been documented. Lamellidens marginalis can thus be regarded as a functional food source as this may combine effectively with other food components for providing essential elements to the body. Moreover, mussel protein hydrolysate provides opportunities for utilizing it in various food formulations and pharmaceuticals. The observations presented herein should be viewed as a prelude to what future holds.

Keywords: functional food, functional properties, Lamellidens marginalis, protein hydrolysate

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2494 Ultrasonic Agglomeration of Protein Matrices and Its Effect on Thermophysical, Macro- and Microstructural Properties

Authors: Daniela Rivera-Tobar Mario Perez-Won, Roberto Lemus-Mondaca, Gipsy Tabilo-Munizaga

Abstract:

Different dietary trends worldwide seek to consume foods with anti-inflammatory properties, rich in antioxidants, proteins, and unsaturated fatty acids that lead to better metabolic, intestinal, mental, and cardiac health. In this sense, food matrices with high protein content based on macro and microalgae are an excellent alternative to meet the new needs of consumers. An emerging and environmentally friendly technology for producing protein matrices is ultrasonic agglomeration. It consists of the formation of permanent bonds between particles, improving the agglomeration of the matrix compared to conventionally agglomerated products (compression). Among the advantages of this process are the reduction of nutrient loss and the avoidance of binding agents. The objective of this research was to optimize the ultrasonic agglomeration process in matrices composed of Spirulina (Arthrospira platensis) powder and Cochayuyo (Durvillae Antartica) flour, by means of the response variable (Young's modulus) and the independent variables were the process conditions (percentage of ultrasonic amplitude: 70, 80 and 90; ultrasonic agglomeration times and cycles: 20, 25 and 30 seconds, and 3, 4 and 5). It was evaluated using a central composite design and analyzed using response surface methodology. In addition, the effects of agglomeration on thermophysical and microstructural properties were evaluated. It was determined that ultrasonic compression with 80 and 90% amplitude caused conformational changes according to Fourier infrared spectroscopy (FTIR) analysis, the best condition with respect to observed microstructure images (SEM) and differential scanning calorimetry (DSC) analysis, was the condition of 90% amplitude 25 and 30 seconds with 3 and 4 cycles of ultrasound. In conclusion, the agglomerated matrices present good macro and microstructural properties which would allow the design of food systems with better nutritional and functional properties.

Keywords: ultrasonic agglomeration, physical properties of food, protein matrices, macro and microalgae

Procedia PDF Downloads 54
2493 Ascidian Styela rustica Proteins’ Structural Domains Predicted to Participate in the Tunic Formation

Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

Abstract:

Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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2492 Sequence Analysis and Structural Implications of Rotavirus Capsid Proteins

Authors: Nishal Parbhoo, John B. Dewar, Samantha Gildenhuys

Abstract:

Rotavirus is the major cause of severe gastroenteritis worldwide in children aged 5 and younger. Death rates are high particularly in developing countries. The mature rotavirus is a non-enveloped triple-layered nucleocapsid containing 11 double-stranded RNA segments. Here a global view on the sequence and structure of the three main capsid proteins, VP7, VP6, and VP2 is taken by generating a consensus sequence for each of these rotavirus proteins, for each species obtained from published data of representative rotavirus genotypes from across the world and across species. The degree of conservation between species was represented on homology models for each of the proteins. VP7 shows the highest level of variation with 14 - 45 amino acids showing conservation of less than 60%. These changes are localized to the outer surface which is exposed to antibodies alluding to a possible mechanism in evading the immune system. The middle layer, VP6 shows lower variability with only 14-32 sites having lower than 70% conservation. The inner structural layer made up of VP2 showed the lowest variability with only 1-16 sites having less than 70% conservation across species. The results correlate with proteins’ multiple structural roles. Although the nucleotide sequences vary due to an error-prone replication and lack of proofreading, the corresponding amino acid sequence of VP2, 6 and 7 remains conserved. Sequence conservation maintained for the virus results in stable protein structures, fit for function. This can be exploited in drug design, molecular studies and biotechnological applications.

Keywords: amino acid sequence conservation, capsid protein, protein structure, vaccine candidate

Procedia PDF Downloads 286
2491 LHCII Proteins Phosphorylation Changes Involved in the Dark-Chilling Response in Plant Species with Different Chilling Tolerance

Authors: Malgorzata Krysiak, Anna Wegrzyn, Maciej Garstka, Radoslaw Mazur

Abstract:

Under constantly fluctuating environmental conditions, the thylakoid membrane protein network evolved the ability to dynamically respond to changing biotic and abiotic factors. One of the most important protective mechanism is rearrangement of the chlorophyll-protein (CP) complexes, induced by protein phosphorylation. In a temperate climate, low temperature is one of the abiotic stresses that heavily affect plant growth and productivity. The aim of this study was to determine the role of LHCII antenna complex phosphorylation in the dark-chilling response. The study included an experimental model based on dark-chilling at 4 °C of detached chilling sensitive (CS) runner bean (Phaseolus coccineus L.) and chilling tolerant (CT) garden pea (Pisum sativum L.) leaves. This model is well described in the literature as used for the analysis of chilling impact without any additional effects caused by light. We examined changes in thylakoid membrane protein phosphorylation, interactions between phosphorylated LHCII (P-LHCII) and CP complexes, and their impact on the dynamics of photosystem II (PSII) under dark-chilling conditions. Our results showed that the dark-chilling treatment of CS bean leaves induced a substantial increase of phosphorylation of LHCII proteins, as well as changes in CP complexes composition and their interaction with P-LHCII. The PSII photochemical efficiency measurements showed that in bean, PSII is overloaded with light energy, which is not compensated by CP complexes rearrangements. On the contrary, no significant changes in PSII photochemical efficiency, phosphorylation pattern and CP complexes interactions were observed in CT pea. In conclusion, our results indicate that different responses of the LHCII phosphorylation to chilling stress take place in CT and CS plants, and that kinetics of LHCII phosphorylation and interactions of P-LHCII with photosynthetic complexes may be crucial to chilling stress response. Acknowledgments: presented work was financed by the National Science Centre, Poland grant No.: 2016/23/D/NZ3/01276

Keywords: LHCII, phosphorylation, chilling stress, pea, runner bean

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2490 Reconfigurable Intelligent Surfaces (RIS)-Assisted Integrated Leo Satellite and UAV for Non-terrestrial Networks Using a Deep Reinforcement Learning Approach

Authors: Tesfaw Belayneh Abebe

Abstract:

Integrating low-altitude earth orbit (LEO) satellites and unmanned aerial vehicles (UAVs) within a non-terrestrial network (NTN) with the assistance of reconfigurable intelligent surfaces (RIS), we investigate the problem of how to enhance throughput through integrated LEO satellites and UAVs with the assistance of RIS. We propose a method to jointly optimize the associations with the LEO satellite, the 3D trajectory of the UAV, and the phase shifts of the RIS to maximize communication throughput for RIS-assisted integrated LEO satellite and UAV-enabled wireless communications, which is challenging due to the time-varying changes in the position of the LEO satellite, the high mobility of UAVs, an enormous number of possible control actions, and also the large number of RIS elements. Utilizing a multi-agent double deep Q-network (MADDQN), our approach dynamically adjusts LEO satellite association, UAV positioning, and RIS phase shifts. Simulation results demonstrate that our method significantly outperforms baseline strategies in maximizing throughput. Lastly, thanks to the integrated network and the RIS, the proposed scheme achieves up to 65.66x higher peak throughput and 25.09x higher worst-case throughput.

Keywords: integrating low-altitude earth orbit (LEO) satellites, unmanned aerial vehicles (UAVs) within a non-terrestrial network (NTN), reconfigurable intelligent surfaces (RIS), multi-agent double deep Q-network (MADDQN)

Procedia PDF Downloads 34
2489 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

Abstract:

GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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2488 Effect of Tensile Strain on Microstructure of Irradiated Core Internal Material

Authors: Hygreeva Kiran Namburi, Anna Hojna, Edita Lecianova, Fencl Zdenek

Abstract:

Irradiation Assisted Stress Corrosion Cracking [IASCC] is one of the most significant environmental degradation in the internal components made from Austenitic stainless steel. This mechanism is still not fully understood and there are no suitable criteria for prediction of the damage during operation. In this work, core basket material 08Ch18N10T austenitic stainless steel acquired from decommissioned NPP Nord / Greifswald Unit 1, VVER 440-230 type, operated for 15 years and irradiated at 5.2 dpa is studied. This material was tensile tested at two different test temperatures and strain rates in air and at the elevated temperature under the water environment. SEM observations of the fracture surface documented ductile fracture of the samples tested in air, but areas of IASCC tested in water. This paper emphasizes on the microscopic examination results from the mechanically tested samples to determine the underlying IASCC physical damage process. TEM observations of thin foils made from the gauge sections that are closer to the fractured surface of the specimen aimed to find variances in interaction of dislocations and grain boundaries owing to different test conditions.

Keywords: irradiation assisted stress corrosion cracking, core basket material, SEM observations of the fracture surface, microscopic examination results

Procedia PDF Downloads 343
2487 Detection of Heroin and Its Metabolites in Urine Samples: A Chemiluminescence Approach

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

Abstract:

A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

Procedia PDF Downloads 258
2486 Using Lysosomal Immunogenic Cell Death to Target Breast Cancer via Xanthine Oxidase/Micro-Antibody Fusion Protein

Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

Abstract:

Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

Procedia PDF Downloads 192
2485 Protein-Thiocyanate Composite as a Sensor for Iron III Cations

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

Abstract:

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

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

Procedia PDF Downloads 415