Search results for: protein matrices

Authors: Aljabryn Dalal Hamad

Abstract:

The present explanatory study concerns with the relation between Diabetes Mellitus and Food Balance in the Kingdom of Saudi Arabia during 2005-2010, using published data. Results illustrated that Saudi citizen daily protein consumption (DPC) during 2005-2007 (g/capita/day) is higher than the average global consumption level of protein with 15.27%, daily fat consumption (DFC) with 24.56% and daily energy consumption (DEC) with 16.93% and increases than recommended level by International Nutrition Organizations (INO) with 56% for protein, 60.49% for fat and 27.37% for energy. On the other hand, DPC per capita in Saudi Arabia decreased during the period 2008-2010 from 88.3 to 82.36 gram/ day. Moreover, DFC per capita in Saudi Arabia decreased during the period 2008-2010 from 3247.90 to 3176.43 Cal/capita/ day, and daily energy consumption (DEC) of Saudi citizen increases than world consumption with 16.93%, while increases with 27.37% than INO. Despite this, DPC, DFC and DEC per capita in Saudi Arabia still higher than world mean. On the other side, results illustrated that the number of diabetic patients in Saudi Arabia during the same period (2005-2010). The curve of diabetic patient’s number in Saudi Arabia during 2005-2010 is regular ascending with increasing level ranged between 7.10% in 2005 and 12.44% in 2010. It is essential to devise Saudi National programs to educate the public about the relation of food balances and diabetes so it could be avoided, and provide citizens with healthy dietary balances tables.

Keywords: Diabetes mellitus, food balance, energy, fat, protein, Saudi Arabia

Procedia PDF Downloads 460

Authors: Gehan Moubarz, Atef M. F. Mohammed, Inas A. Saleh, Heba Mahdy-Abdallah, Amal Saad-Hussein

Abstract:

This study estimates the association between respiratory outcomes among employees of a secondary aluminum plant and airborne pollutants. Additionally, it looks into the relationship between pulmonary dysfunction in workers and XRCC1 gene polymorphisms. 110 exposed workers and 58 non-exposed workers participated in the study. Measurements have been conducted on SO₂, NO₂, and particulate particles. Pulmonary function was tested. Eosinophil cationic protein (ECP), C-reactive protein (CRP), matrix metalloproteinase-1 (MMP-1), interleukin 6 (IL6), GM-CSF, X-Ray Repair Cross Complementing 1 (XRCC1) protein, and genotyping of XRCC1 gene polymorphisms were examined. Results: The annual average concentrations of (PM₂.₅, PM₁₀, TSP, SO₂, and NO₂) were lower than the permissible limit. The areas around ovens, evaporators, and cold rolling mills exhibited the highest amounts. The majority of employees in these departments had impaired lung function. With longer exposure times, the exposed group's FEV1% and FVC% considerably reduced. The exposed workers had considerably higher XRCC1 levels. The evaluated inflammatory biomarkers showed no statistically significant difference. Conclusion: Aluminum workers are at risk of developing respiratory disorders. The level of serum XRCC1 may act as a biomarker that might be very useful for detecting susceptible workers.

Keywords: aluminum industry, particulate matter, SO₂, NO₂, lung function, XRCC1 gene polymorphism, XRCC1 protein, inflammatory biomarkers

Procedia PDF Downloads 10

Authors: Jiang Xu, Daoping Wang, Yinghong Pan

Abstract:

The jointing-booting stage is a critical period of both vegetative growth and reproductive growth in rice. Therefore, the proteomic analysis of the mutant Osbri1, whose corresponding gene OsBRI1 encodes the putative BRs receptor OsBRI1, at jointing-booting stage is very important for understanding the effects of BRs on vegetative and reproductive growth. In this study, the proteomes of leaves from an allelic mutant of the DWARF 61 (D61, OsBRI1) gene, Fn189 (dwarf54, d54) and its wild-type variety T65 (Taichung 65) at jointing-booting stage were analysed by using a Q Exactive plus orbitrap mass spectrometer, and more than 3,100 proteins were identified in each sample. Ontology analysis showed that these proteins distribute in various space of the cells, such as the chloroplast, mitochondrion, and nucleus, they functioned as structural components and/or catalytic enzymes and involved in many physiological processes. Moreover, quantitative analysis displayed that 266 proteins were differentially expressed in two samples, among them, 77 proteins decreased and 189 increased more than two times in Fn189 compared with T65, the proteins whose content decreased in Fn189 including b5-like Heme/Steroid binding domain containing protein, putative retrotransposon protein, putative glutaminyl-tRNA synthetase, and higher content proteins such as mTERF, putative Oligopeptidase homologue, zinc knuckle protein, and so on. A former study founded that the transcription level of a mTERF was up-regulated in the leaves of maize seedling after EBR treatment. In our experiments, it was interesting that one mTERF protein increased, but another mTERF decreased in leaves of Fn189 at jointing-booting stage, which suggested that BRs may have differential regulation mechanisms on the expression of various mTERF proteins. The relationship between other differential proteins with BRs is still unclear, and the effects of BRs on rice protein contents and its regulation mechanisms still need further research.

Keywords: bri1 mutant, jointing-booting stage, proteomic analysis, rice

Procedia PDF Downloads 246

Authors: Afiqah Shafify Amran, Siti Aisyah Shamsudin, Nurul Yuziana Mohd Yusof

Abstract:

Cadmium Sulphide (CdS) from group II-IV quantum dots with good optical properties was successfully synthesized by using the simple colloidal method. Capping them with ligand Polyethylinamine (PEI) alters the surface defect of CdS while, thioglycolic acid (TGA) was added to the reaction as a stabilizer. Due to their cytotoxicity, we decided to conjugate them with the protein cage nanoparticles. In this research, we used capsid of Cowpea Chlorotic Mottle Virus (CCMV) to package the CdS because they have the potential to serve in drug delivery, cell targeting and imaging. Adding Sodium Hydroxide (NaOH) changes the pH of the systems hence the isoelectric charge is adjusted. We have characterized and studied the morphology and the optical properties of CdS and CdS-CCMV by transmitted electron microscopic (TEM), UV-Vis spectroscopy, photoluminescence spectroscopy, UV lamp and Fourier transform infrared spectroscopy (FTIR), respectively. The results obtained suggest that the protein cage nanoparticles do not affect the optical properties of CdS.

Keywords: cadmium sulphide, cowpea chlorotic mottle virus, protein cage nanoparticles, quantum dots

Procedia PDF Downloads 338

Authors: Abdelsabour G. A. Khaled, Ahmed W. A. Abdalla And Sabry Y. M. Mahmoud

Abstract:

Banana plants showing typical mosaic and yellow stripes on leaves as symptoms were collected from Assiut Governorate in Egypt. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of symptoms, transmission, serology, transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Coat protein (CP) gene was amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. In this study the CMV was transmitted into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA). Analysis of the 120 deduced amino acid sequence of the coat protein gene revealed that the EG-A strain of CMV shared from 97.50 to 98.33% with those strains belonging to subgroup IA. The cluster analysis grouped the Egyptian isolate with strains Fny and Ri8 belonging sub-group IA. It appears that there occurs a high incidence of CMV infecting banana belonging to IA subgroup in most parts of Egypt.

Keywords: banana, CMV, transmission, CP gene, RT-PCR

Procedia PDF Downloads 341

Authors: Saha Saradindu, Das Payel, Somdeb BoseDasgupta

Abstract:

Tuberculosis caused by Mycobacteria tuberculosis is a dreadful disease and more so with the advent of extreme and total drug-resistant species. Mycobacterial pathogenesis is an ever-changing paradigm from phagosome maturation block to phagosomal escape into macrophage cytosol and finally acid tolerance and survival inside the lysosome. Mycobacteria are adept at subverting the host immune response by highjacking host cell signaling and secreting virulence factors. One such virulence factor is a ser/thr kinase; Protein kinase G (PknG), which is known to prevent phagosome maturation. The host substrates of PknG, allowing successful pathogenesis still remain an enigma. Hence we carried out a comparative phosphoproteomic screen and identified a number of substrates phosphorylated by PknG. We characterized some of these substrates in vivo and in vitro and observed that PknG mediated phosphorylation of these substrates leads to reduced TNFa production as well as decreased response to TNFa induced macrophage necroptosis, thus enabling mycobacterial survival and proliferation.

Keywords: mycobacteria, Protein kinase G, phosphoproteomics, necroptosis

Procedia PDF Downloads 146

Authors: V. Agostini, S. Gino, S. Inturri, A. Piccinini

Abstract:

In cases of corpse identification or parental testing performed on exhumed alleged dead father, usually, we seek and acquire organic samples as bones and/or bone fragments, teeth, nails and muscle’s fragments. The DNA analysis of these cadaveric matrices usually leads to identifying success, but it often happens that the results of the typing are not satisfactory with highly degraded, partial or even non-interpretable genetic profiles. To aggravate the interpretative panorama deriving from the analysis of such 'classical' organic matrices, we must add a long and laborious treatment of the sample that starts from the mechanical fragmentation up to the protracted decalcification phase. These steps greatly increase the chance of sample contamination. In the present work, instead, we want to report the use of 'unusual' cadaveric matrices, demonstrating that their forensic genetics analysis can lead to better results in less time and with lower costs of reagents. We report six case reports, result of on-field experience, in which eyeswabs and cartilage were sampled and analyzed, allowing to obtain clear single genetic profiles, useful for identification purposes. In all cases we used the standard DNA tissue extraction protocols (as reported on the user manuals of the manufacturers such as QIAGEN or Invitrogen- Thermo Fisher Scientific), thus bypassing the long and difficult phases of mechanical fragmentation and decalcification of bones' samples. PCR was carried out using PowerPlex® Fusion System kit (Promega), and capillary electrophoresis was carried out on an ABI PRISM® 310 Genetic Analyzer (Applied Biosystems®), with GeneMapper ID v3.2.1 (Applied Biosystems®) software. The software Familias (version 3.1.3) was employed for kinship analysis. The genetic results achieved have proved to be much better than the analysis of bones or nails, both from the qualitative and quantitative point of view and from the point of view of costs and timing. This way, by using the standard procedure of DNA extraction from tissue, it is possible to obtain, in a shorter time and with maximum efficiency, an excellent genetic profile, which proves to be useful and can be easily decoded for later paternity tests and/or identification of human remains.

Keywords: DNA, eye swabs and cartilage, identification human remains, paternity testing

Procedia PDF Downloads 109

Authors: Khushi Kashyap, Pratibha Singh

Abstract:

In the recent years increasing interest in vegetarian diets has been observed, a major problem in this type of diet is to provide the appropriate amount of protein .Value addition of food is current most talked topic because of increasing nutritional awareness among consumers today. An investigation was conducted to develop protein rich multi-millet hemp seed khakhra. The seeds of cannabis sativa L. have been a significant source of food for thousand of year. In recent years, hemp has not been thoroughly explored for its nutritional potential due to the mistaken belief regarding the cannabis plants. Methodology- two variations was prepared referencing standard recipe. Variation 1 was prepared using 25g ragi, 25g bajra,40g whole wheat flour with 10g hemp seed powder, variation 2(RF-25g,BF25g,WWF-35g,HS-15g). The product was subjected to sensory evolution by semi trained panel members using 9 point hedonic on 50 panelists. Result- result of the sensory evaluation revealed that the product incorporated with 15g of hemp seed were similar to control I texture, taste and overall quality and was more acceptable by the panelist and was selected as final product seed. On estimation of the nutrient content 30g of khakhra provides 107kcal of energy,12g protein,75g carbohydrate, and 9.6g of fats with shelf life of 3 months. Conclusion- khakhras can be eaten as a snack at any time of the day. hemp seed powder incorporated in it enhances its nutritive value and makes it more nutritious. It is suitable for consumption of all the age group.

Keywords: cannabis sativa, hemp, protein, seed

Procedia PDF Downloads 90

Authors: Khurram Munir, Cuie Wen, Yuncang Li

Abstract:

Magnesium (Mg) metal matrix composites (MMCs) reinforced with graphene nanoplatelets (GNPs) have been developed by powder metallurgy (PM). In this study, GNPs with different concentrations (0.1-0.3 wt.%) were dispersed into Mg powders by high-energy ball-milling processes. The microstructure and resultant mechanical properties of the fabricated nanocomposites were characterized using transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Raman spectroscopy (RS), compression and nano-wear tests. The corrosion resistance of the fabricated composites was evaluated by electrochemical tests and hydrogen evolution measurements. Finally, the biological response of Mg-GNPs composites was assessed using osteoblast-like SaOS2 cells. The results indicate that GNPs are excellent candidates as reinforcements in Mg matrices for the manufacture of biodegradable Mg-based composite implants. GNP addition improved the mechanical properties of Mg via synergetic strengthening modes. Moreover, retaining the structural integrity of GNPs during PM processing improved the ductility, compressive strength, and corrosion resistance of the Mg-GNP composites as compared to monolithic Mg. Cytotoxicity assessments did not reveal any significant toxicity with the addition of GNPs to Mg matrices. This study demonstrates that Mg-xGNPs with x < 0.3 wt.%, may constitute novel biodegradable implant materials for load-bearing applications.

Keywords: magnesium-graphene composites, strengthening mechanisms, In vitro cytotoxicity, biocorrosion

Procedia PDF Downloads 158

Authors: Hung-Chieh Chen, Kuo-Hui Wang, Tsu-Lin Yeh

Abstract:

Proteomics represent the performance of genomic complement proteins and the protein level on functional genomics. This study adopted proteomic strategies for comparing serum proteins among three groups: elite sprinter (sprint runner group, SR), long-distance runners (long-distance runner group, LDR), and the untrained control group (control group, CON). Purposes: This study aims to identify elite sprinters and long-distance runners’ serum protein and to provide a comparison of their serum proteome’ composition. Methods: Serum protein fractionations that separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and analyzed by a quantitative nano-LC-MS/MS-based proteomic profiling. The one-way analysis of variance (ANOVA) and Scheffe post hoc comparison (α= 0.05) was used to determine whether there is any significant difference in each protein level among the three groups. Results: (1) After analyzing the 307 identified proteins, there were 26 unique proteins in the SR group, and 18 unique proteins in the LDR group. (2) For the LDR group, 7 coagulation function-associated proteins’ expression levels were investigated: vitronectin, serum paraoxonase/arylesterase 1, fibulin-1, complement C3, vitamin K-dependent protein, inter-alpha-trypsin inhibitor heavy chain H3 and von Willebrand factor, and the findings show the seven coagulation function-associated proteins were significantly lower than the group of SR. (3) Comparing to the group of SR, this study found that the LDR group’s expression levels of the 2 antioxidant proteins (afamin and glutathione peroxidase 3) were also significantly lower. (4) The LDR group’s expression levels of seven immune function-related proteins (Ig gamma-3 chain C region, Ig lambda-like polypeptide 5, clusterin, complement C1s subcomponent, complement factor B, complement C4-A, complement C1q subcomponent subunit A) were also significantly lower than the group of SR. Conclusion: This study identified the potential serum protein markers for elite sprinters and long-distance runners. The changes in the regulation of coagulation, antioxidant, or immune function-specific proteins may also provide further clinical applications for these two different track athletes.

Keywords: biomarkers, coagulation, immune response, oxidative stress

Procedia PDF Downloads 117

Authors: Olaide Ruth Aderibigbe, Oluwatoyin Oluwole

Abstract:

Pigeon pea is one of the minor leguminous plants; though underutilised, it is used traditionally by farmers to alleviate hunger and malnutrition. Pigeon pea is cultivated in Nigeria by subsistence farmers. It is rich in protein and minerals, however, its utilisation as food is only common among the poor and rural populace who cannot afford expensive sources of protein. One of the factors contributing to its limited use is the high antinutrient content which makes it indigestible, especially when eaten by children. The development of value-added products that can reduce the antinutrient content and make the nutrients more bioavailable will increase the utilisation of the crop and contribute to reduction of malnutrition. This research, therefore, determined the effects of different roasting temperatures (130 0C, 140 0C, and 150 0C) on the proximate, mineral and antinutrient component of a pigeon pea snack. The brown variety of pigeon pea seeds were purchased from a local market- Otto in Lagos, Nigeria. The seeds were cleaned, washed, and soaked in 50 ml of water containing sugar and salt (4:1) for 15 minutes, and thereafter the seeds were roasted at 130 0C, 140 0C, and 150 0C in an electric oven for 10 minutes. Proximate, minerals, phytate, tannin and alkaloid content analyses were carried out in triplicates following standard procedures. The results of the three replicates were polled and expressed as mean±standard deviation; a one-way analysis of variance (ANOVA) and the Least Significance Difference (LSD) were carried out. The roasting temperatures significantly (P<0.05) affected the protein, ash, fibre and carbohydrate content of the snack. Ready-to-eat snack prepared by roasting at 150 0C significantly had the highest protein (23.42±0.47%) compared the ones roasted at 130 0C and 140 0C (18.38±1.25% and 20.63±0.45%, respectively). The same trend was observed for the ash content (3.91±0.11 for 150 0C, 2.36±0.15 for 140 0C and 2.26±0.25 for 130 0C), while the fibre and carbohydrate contents were highest at roasting temperature of 130 0C. Iron, zinc, and calcium were not significantly (P<0.5) affected by the different roasting temperatures. Antinutrients decreased with increasing temperature. Phytate levels recorded were 0.02±0.00, 0.06±0.00, and 0.07±0.00 mg/g; tannin levels were 0.50±0.00, 0.57±0.00, and 0.68±0.00 mg/g, while alkaloids levels were 0.51±0.01, 0.78±0.01, and 0.82±0.01 mg/g for 150 0C, 140 0C, and 130 0C, respectively. These results show that roasting at high temperature (150 0C) can be utilised as a processing technique for increasing protein and decreasing antinutrient content of pigeon pea.

Keywords: antinutrients, pigeon pea, protein, roasting, underutilised species

Procedia PDF Downloads 141

Authors: Yue He, Youn Young Shim, Ji Hye Kim, Jae Youl Cho, Martin J. T. Reaney

Abstract:

Recently, pulse cooking water (a.k.a. Aquafaba) has been used as an important and cost-effective alternative to eggs in gluten-free, vegan cooking and baking applications. The aquafaba (AQ) is primarily due to its excellent ability to stabilize foams and emulsions in foods. However, the functional ingredients of this excellent AQ are usually discarded with the compound release. This study developed a high-functional food material, AQ, using functional soybean AQ that has not been studied in Korea. A zero-waste and cost-effective hybrid process were used to produce oil emulsifiers from Korean soybeans. The treatment technique was implemented using a small number of efficient steps. Aquafaba from Backtae had the best emulsion properties (92%) and has the potential to produce more stable food oil emulsions. Therefore, this study is expected to be utilized in the development of the first gluten-free, vegan product for vegetarians and consumers with animal protein allergies, utilizing wastewater from cooked soybeans as a source of plant protein that can replace animal protein.

Keywords: aquafaba, soybean, chickpea, emulsifiers, egg replacer, egg-free products

Procedia PDF Downloads 176

Authors: Jong Il Rhee

Abstract:

The polypeptide representing the mature part of human BMP-7 was cloned and efficiently expressed in Escherichia coli and Bacillus subtilis, which had a clear band for hBMP-7, a homodimeric protein with an apparent molecular weight of 15.4 kDa. Recombinant E.coli produced 111 pg hBMP-7/mg of protein hBMP-7 through IPTG induction. Recombinant B. subtilis also produced 350 pg hBMP-7/ml of culture medium. The hBMP-7 was purified in 2 steps using an FPLC system with an ion exchange column and a gel filtration column. The hBMP-7 produced in this work also stimulated the alkaline phosphatase (ALP) activity in a dose-dependent manner, i.e. 2.5- and 8.9-fold at 100 and 300 ng hBMP-7/ml, respectively, and showed intact biological activity.

Keywords: B. subtilis, E. coli, fermentation, hBMP-7

Procedia PDF Downloads 441

Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

Abstract:

We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

Procedia PDF Downloads 227

Authors: Elena K. Schneider, Johnny X. Huang, Vincenzo Carbone, Mark Baker, Mohammad A. K. Azad, Matthew A. Cooper, Jian Li, Tony Velkov

Abstract:

Ivacaftor is a novel CF trans-membrane conductance regulator (CFTR) potentiator that improves the pulmonary function for cystic fibrosis patients bearing a G551D CFTR-protein mutation. Because ivacaftor is highly bound (>97%) to plasma proteins, there is the strong possibility that co-administered CF drugs that compete for the same plasma protein binding sites and impact the free drug concentration. This in turn could lead to drastic changes in the in vivo efficacy of ivacaftor and therapeutic outcomes. This study compares the binding affinity of ivacaftor and co-administered CF drugs for human serum albumin (HSA) and α1-acid glycoprotein (AGP) using surface plasmon resonance and fluorimetric binding assays that measure the displacement of site selective probes. Due to their high plasma protein binding affinities, drug-drug interactions between ivacaftor are to be expected with ducosate, montelukast, ibuprofen, dicloxacillin, omeprazole and loratadine. The significance of these drug-drug interactions is interpreted in terms of the pharmacodynamic/pharmacokinetic parameters and molecular docking simulations. The translational outcomes of the data are presented as recommendations for a staggered treatment regimen for future clinical trials which aims to maximize the effective free drug concentration and clinical efficacy of ivacaftor.

Keywords: human α-1-acid glycoprotein, binding affinity, human serum albumin, ivacaftor, cystic fibrosis

Procedia PDF Downloads 308

Authors: Thi Thao Ninh, Yuri Trusov, José Ramón Botella

Abstract:

Heterotrimeric G proteins, comprised of three subunits, α, β and γ, are involved in signal transduction pathways that mediate a vast number of processes across the eukaryotic kingdom. 23 Gα subunits are present in humans whereas most plant genomes encode for only one canonical Gα. The disparity observed between Arabidopsis, rice, and maize Gα-deficient mutant phenotypes suggest that Gα functions have diversified between eudicots and monocots during evolution. Alternatively, since the only Gα mutations available in dicots have been produced in Arabidopsis, the possibility exists that this species might be an exception to the rule. In order to test this hypothesis, we studied the G protein α subunit (TGA1) in tomato. Four tga1 knockout lines were generated in tomato cultivar Moneymaker using CRISPR/Cas9. The tga1 mutants exhibit a number of auxin-related phenotypes including changes in leaf shape, reduced plant height, fruit size and number of seeds per fruit. In addition, tga1 mutants have increased sensitivity to abscisic acid during seed germination, reduced sensitivity to exogenous auxin during adventitious root formation from cotyledons and excised hypocotyl explants. Our results suggest that Gα mutant phenotypes in tomato are very similar to those observed in monocots, i.e. rice and maize, and cast doubts about the validity of using Arabidopsis as a model system for plant G protein studies.

Keywords: auxin-related phenotypes, CRISPR/Cas9, G protein α subunit, heterotrimeric G proteins, tomato

Procedia PDF Downloads 136

Authors: Ashima Sharma

Abstract:

Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

Procedia PDF Downloads 209

Authors: Chaoran Liu

Abstract:

Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

Procedia PDF Downloads 233

Authors: Mobil Kazimov, Guseyn İbragimov

Abstract:

InAs-CrAs systems are synthesized by the vertical Bridgman–Stockbarger method. XRD analysis and microstructural study of InAs-CrAs composites show that CrAs metallic inclusions are uniformly distributed in the InAs matrices.

Keywords: XRD, eutectic alloy, SEM, EDX

Procedia PDF Downloads 76

Authors: Tagouelbe Tiho, Amissa Augustin Adima, Yao Casimir Brou, Nabayo Traore, Gouha Firmin Kouassi, Thierry Roland Kouame, Maryline Kouba

Abstract:

The work aimed to study Borassus aethiopum Mart (B.a) dried pulp nutritional value for its incorporation in human and poultry diets. Firstly, the mature fruit macro-composition was assessed. Secondly, the pulp was dried at 40, 50, 60, 70, and 80ᵒC. Thereafter, the analysis was performed for fat, protein, total sugars, Ca, P, Mg, and fatty acid profile monitoring. As a result, the fruits weighed 1,591.35, delivered 516.73, and 677.82 grams of pulp and seeds, respectively. Mainly, increasing heat adversely affected the outputs. Consequently, the fat results were 14.12, 12.97, 8.93, 8.89ᶜ, and 5.56%; protein contents were 11.64, 10.15, 8.97, 8.84, and 8.42%; total sugar deliveries were 6.28, 6.05, 5.26, 5.02, and 4.76% (P < 0.01). Thereafter, the metabolizable energies were 3,785.22; 3,834.28; 3,616.62; 3,667.03; and 3,608.33 kcal/kg (DM). Additionally, Calcium (Ca) contents were 0.51, 0.55, 0.69, 0.77, and 0.81%, while phosphorus (P) mean was 0.17%, and the differences were not significant (P < 0.01). So, the Ca/P ratios were 2.79, 3.04, 4.10, 4.71, and 4.95. Finally, fatty acids (FA) assessments revealed 22.33 saturated (SFA), 77.67 unsaturated (UFA), within which 67.59% were monounsaturated (MUFA). Interestingly, the rising heat depressed n-6/n-3 ratios that were 1.1, 1.1, 0.45 and 0.38, respectively at 40, 50, 70 and 80ᵒC. In short, drying did not only enhance the product shelf life but it also improved the nutritional value. Thus, B.a mature fruit pulps dried at 70ᵒC are good functional foods, with more than 66% MUFA, and energy source for human and poultry nutrition.

Keywords: Borassus aethiopum Mart, fatty acids, metabolizable energy, minerals, protein

Procedia PDF Downloads 171

Authors: Hamad Rafique

Abstract:

The bioactive peptide RDFPITWPW (RW-9) identified from oat protein has been reported to be positive in memory deficits. However, no clarity on the mechanisms responsible for the neuroprotective effects of RW-9 peptide against AD-like symptoms. Herein, it found that RW-9 intervention showed various improving effects in cognitive-behavioral tests and alleviated oxidative stress and inflammation in the scopolamine-induced mice model. The hippocampus proteomics analysis revealed the upregulation of memory-related proteins, including Grin3a, Ppp2r1b, Stat6, Pik3cd, Slc5a7, Chrm2, mainly involved in cAMP signaling, PI3K-Akt signaling, and JAK-STAT signaling pathways. The administration of RW-9 significantly upregulated the neurotransmitters, including 5-HT, DA, and Arg, in mice brains. Moreover, it regulated the serum metabolic profile and increased the expression levels of ABC transporters, biosynthesis of amino acids, and Amino acyl-tRNA biosynthesis, among others. The 16s-rRNA results illustrated that the RW-9 restored the abundance of Muribaculaceae, Lachnospiraceae, Lactobacillus, Clostridia and Bactericides. Taken together, our results suggest that the RW-9 may prevent the AD-like symptoms via modulation of the gut-serum-brain axis.

Keywords: oat protein, active peptide, neuroprotective, gut-brain axis

Procedia PDF Downloads 27

Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan

Abstract:

The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson, and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence-based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation-prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: aggregation, amyloids, thermophilic proteins, amino acid residues, machine learning techniques

Procedia PDF Downloads 614

Authors: Natela Gerliani, Mohammed Aider

Abstract:

Nowadays, the interest of using sustainable technologies for protein extraction from underutilized oilseeds is growing. Currently, a major disposal problem for the oil industry is by-products of plant food processing such as soybean meal. That is why valorization of soybean meal is important for the oil industry since it contains high-quality proteins and other valuable components. Generally, soybean meal is used in livestock and poultry feed but is rarely used in human feed. Though chemical composition of this meal compensate nutritional deficiency and can be used to balance protein in human food. Regarding the efficiency of soybean meal valorization, extraction is a key process for obtaining enriched protein ingredient, which can be incorporated into the food matrix. However, most of the food components such as proteins extracted from oilseeds by-products imply the utilization of organic and inorganic chemicals (e.g. acids, bases, TCA-acetone) having a significant environmental impact. In a context of sustainable production, the use of an electro-activation technology seems to be a good alternative. Indeed, the electro-activation technology requires only water, food grade salt and electricity as main materials. Moreover, this innovative technology helps to avoid special equipment and trainings for workers safety as well as transport and storage of hazardous materials. Electro-activation is a technology based on applied electrochemistry for the generation of acidic and alkaline solutions on the basis of the oxidation-reduction reactions that occur at the vicinity electrode/solution interfaces. It is an eco-friendly process that can be used to replace the conventional acidic and alkaline extraction. In this research, the electro-activation technology for protein extraction from soybean meal was carried out in the electro-activation reactor. This reactor consists of three compartments separated by cation and anion exchange membranes that allow creating non-contacting acidic and basic solutions. Different current intensities (150 mA, 300 mA and 450 mA) and treatment durations (10 min, 30 min and 50 min) were tested. The results showed that the extracts obtained by the electro-activation method have good quality in comparison to conventional extracts. For instance, extractability obtained with electro-activation method was 55% whereas with the conventional method it was only 36%. Moreover, a maximum protein quantity of 48 % in the extract was obtained with the electro-activation technology comparing to the maximum amount of protein obtained by conventional extraction of 41 %. Hence, the environmentally sustainable electro-activation technology seems to be a promising type of protein extraction that can replace conventional extraction technology.

Keywords: by-products, eco-friendly technology, electro-activation, soybean meal

Procedia PDF Downloads 228

Authors: Abdu Umar Adamu, Saidu Akun Abdullahi, Al-Hassan Muhammed, Hamisu Abdu

Abstract:

Leguminous plants such as beans have been considered as a source of protein in this present work. The proximate analysis on beans (Vigna unguiculata) were determined in order to identify the nutritional content as well as presence of some heavy metals accumulation in washed and unwashed beans (white Beans) sold in Kazaure market Jigawa State Nigeria. On the average comparative analysis, the result has indicated that, the Vigna unguiculata had protein content of 61.1%, fibre 4.5%, ash 10.4%, moisture 5%, carbohydrate 15.8% and total lipid 4.9%, therefore it could be suggested that beans has enough nutritional content that helps the people health. The heavy metal analysis of unwashed white beans showed that Fe (17.37 ± 6.71)mg/kg had the highest concentration followed by Zn (6.41 ± 3.09), Cu (5.69 ± 2.42), Cd (0.46 ± 0.65) and Pb (0.57 ± 0.94)mg/kg , while the washed beans shows that Zn (0.11 ± 0.17), Fe (0.01 ± 0.006), Cd (0.02 ± 0.01), Cu (0.03 ± 0.021), Pb (0.01 ± 0.006)mg/kg. The washed white beans are safe for consumption and also the concentration of heavy metal are negligible and of nontoxic effect to human health.

Keywords: white beans, protein, proximate composition, heavy metal

Procedia PDF Downloads 432

Authors: Ruolan Zhou

Abstract:

Trisomy of human chromosome 21 is the primary cause of Down Syndrome (DS), and this genetic disease has significantly burdened families and countries, causing great controversy. To address this problem, the research takes an approach in exploring the relationship between genetic abnormality and this disease's symptoms, adopting several techniques, including data analysis and enrichment analysis. It also explores open-source websites, such as NCBI, DAVID, SOURCE, STRING, as well as UCSC, to complement its result. This research has analyzed the variety of genes on human chromosome 21 with simple coding, and by using analysis, it has specified the protein-coding genes, their function, and their location. By using enrichment analysis, this paper has found the abundance of keratin production-related coding-proteins on human chromosome 21. By adopting past researches, this research has attempted to disclose the relationship between trisomy of human chromosome 21 and keratin production abnormality, which might be the reason for common diseases in patients with Down Syndrome. At last, by addressing the advantage and insufficiency of this research, the discussion has provided specific directions for future research.

Keywords: Down Syndrome, protein production, genome, enrichment analysis

Procedia PDF Downloads 126

Authors: Anjum-Zubair, Muhammad, Muhammad Shoaib Alam, Muhammad Samee Mubarik, Iftikhar Ahmad

Abstract:

The aim of present research was to evaluate the effect of dietary protein (soybean) formulated feed on the growth performance of carp fish seed (Rohu, Mori, Grass, and Gulfam) in ponds under polyculture system. Keeping in view the protein requirements of these four carps, they were fed with formulated feed contains 30% of crude protein. The fingerlings were fed once on daily basis at 5% of their wet body weight. A 90 days experiment was conducted in two cemented ponds situated at Fish Seed Hatchery and Research Centre, Rawal Town, Islamabad, Pakistan. Pond1 contain major carps i.e. Rohu and Mori while pond 2 was stocked with Chinese carps i.e. Grass carp and Gulfam. Random sampling of five individuals of each species was done fortnightly to measure the body weight and total body length. Maximum growth was observed in fingerling of Grass carp followed by Mori, Rohu and Gulfam. Total fish production was recorded as Grass 623.45 gm followed by Mori 260.3 gm, Rohu 243.08 gm and Gulfam 181.165 gm respectively. Significantly results were obtained among these four fish species when the corresponding data was subjected to statistical analysis by using two sample t-test. The survival rate was 100%. Study shows that soybean as plant based protein can be easily used as substitute to fish meal without any adverse effect on fish health and fish production.

Keywords: carps, fry growth, poly culture, soybean meal

Procedia PDF Downloads 500

Authors: A. Ojha, Y. K. Gupta

Abstract:

Organophosphate (OP) pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests throughout the world. Chlorpyrifos (CPF), methyl parathion (MPT), and malathion (MLT) are among the most extensively used OP pesticides in India. DNA strand breaks and DNA-protein crosslinks (DPC) are toxic lesions associated with the mechanisms of toxicity of genotoxic compounds. In the present study, we have examined the potential of CPF, MPT, and MLT individually and in combination, to cause DNA strand breakage and DPC formation. Peripheral blood lymphocytes of rat were exposed to 1/4 and 1/10 LC50 dose of CPF, MPT, and MLT for 2, 4, 8, and 12h. The DNA strand break was measured by the comet assay and expressed as DNA damage index while DPC estimation was done by fluorescence emission. There was significantly marked increase in DNA damage and DNA-protein crosslink formation in time and dose dependent manner. It was also observed that MPT caused the highest level of DNA damage as compared to other studied OP compounds. Thus, from present study, we can conclude that studied pesticides have genotoxic potential. The pesticides mixture does not potentiate the toxicity of each other. Nonetheless, additional in vivo data are required before a definitive conclusion can be drawn regarding hazard prediction to humans.

Keywords: organophosphate, pesticides, DNA damage, DNA protein crosslink, genotoxic

Procedia PDF Downloads 356

Authors: Izzatul Ulfana, Angga Pratomo Cahyadi, Rimayanti, Widjiati

Abstract:

Freezing oocyte could cause temperature stress. Temperature stress triggers cell damage. Insulin Transferrin Selenium (ITS) and Heat Shock Protein 70 (HSP70) had been used to prevent damage to the oocyte after freezing. ITS and HSP70 could cause the difference protective effect. The aim of this research was to obtain an effective cryoprotectant for freezing local goat oocyte in cumulus cells change. The research began by collecting the ovary from a local slaughterhouse in Indonesia, aspiration follicle, in vitro maturation and the freezing had been used vitrification method. Examination of the morphology cells by native staining method. Data on the calculation morphology oocyte analyzed by Kruskall-Wallis Test. After the Kruskall-Wallis Test which indicated significance, followed by Mann-Whitney Test to compare between treatment groups. As a result, cryoprotectant ITS has the best culumus cells after warming

Keywords: Insulin Transferrin Selenium, Heat Shock Protein 70, cryoprotectant, vitrification

Procedia PDF Downloads 236

Authors: Jiraporn Rojtinnakorn

Abstract:

ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I trans-membrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish.

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor

Procedia PDF Downloads 226

Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

Abstract:

A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

Procedia PDF Downloads 271