Search results for: synthetic protein design
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 15078

Search results for: synthetic protein design

14928 The Role of Flowering Pesticidal Plants for Sustainable Pest Management

Authors: Baltazar Ndakidemi

Abstract:

The resource-constrained farmers, especially those in sub-Saharan Africa, encounter significant challenges related to agriculture, notably diseases and pests. The sustainable means of pest management are not well known to farmers. As a result, some farmers use synthetic pesticides whose environmental impacts, ill health, and other negative impacts of synthetic pesticides on natural enemies have posed a great need for more sustainable means of pest management. Pesticidal plant resources can replace synthetic pesticides because their secondary metabolites can exhibit insecticidal activities such as deterrence, repellence, and pests' mortality. Additionally, the volatiles from these plants can have positive effects of attracting populations of natural enemies. Pesticidal plants can be grown as field margin plants or in strips for supporting natural enemies' populations. However, this is practically undetermined. Hence, there is a need to investigate the roles played by pesticidal plants in supporting natural enemies of pests and their applications in different cropping systems such as legumes. This study investigates different pesticidal plants with a high potential for pest control in agricultural fields. The information sheds light on potential plants that can be used for different crop pests.

Keywords: natural enemies, biological control, synthetic pesticides, pesticidal plants, predators, parasitoids

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14927 Effect of Supplemental Phytase on the Digestibility of Crude Protein and Phosphorus of Rice Husk in Broiler Chicken

Authors: Ibinabo I. Ilaboya, Eustace A. Iyayi

Abstract:

Phosphorus (P) is an indispensable mineral in broiler diets. Rice husk contains phytate-P and other nutrients like protein, carbohydrates, which are poorly digested in broiler chickens. Broiler chickens (BC) lacks sufficient phytase to help hydrolyse phytate-bound P. Hence excess of P is excreted by these chickens into the environment causing environmental pollution. Supplementation of such diets with microbial phytase helps to improve the digestibility of these nutrients. The study was conducted to determine the effect of phytase supplementation on the digestibility of crude protein (CP) and P of rice husk in BC. Six semi-purified diets of three levels of total P (3.46, 4.91 and 6.37g/kg) without and with 1,000 units of phytase per kg were formulated. Titanium dioxide was added to the diets at the rate of 5g/kg as an indigestible marker. At 20dposthatch, 288 broilers (Abor Acre) were weighed and allotted to the diets with 6 replicates of 8 birds each in a randomized complete block design. The birds had free access to the experimental diets until day 26 post-hatch. Phytase supplementation increased (p < 0.05) digestibility of P from 75-93%. Rice husk and its interaction with phytase had no significant (p > 0.05) effect on P digestibility, whereas there was significant (p < 0.01) effect on the interaction of rice husk with phytase on CP digestibility. There were linear increases (p < 0.01) in digested P and CP with phytase supplementation. The P and CP losses from the BC was reduced with the addition of phytase. Results suggest that supplementation of rice husk-based diets with microbial phytase improved pre-caecal digestibility of P and CP in broilers.

Keywords: crude protein, phosphorus, phytase, rice husk

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14926 Chitin Nanocrystals as Sustainable Surfactant Alternative for Enhancing Oil-in-Water Emulsions Stability in Oil and Gas Fields

Authors: A. Altomi, A. Alhebshi, M. Rasm, B. Osman

Abstract:

This study explored the application of chitin nanocrystals (ChiNCs), derived from a renewable and environmentally friendly material, as stabilizers for oil-in-water (O/W) emulsions. O/W emulsions are commonly used in various applications but are prone to instability and degradation over time. Instability can occur due to factors such as flocculation, coalescence, and gravitational separation, including creaming and sedimentation, either independently or simultaneously. To produce ChiNCs, chitin powder underwent acid hydrolysis. Transmission electron microscopy (TEM) analysis revealed that ChiNCs exhibited a needle-like morphology, with lengths ranging from 200 to 800 nm and widths ranging from 20 to 80 nm. The surface charge of ChiNCs was negative at pH values above 7 and positive at pH values below 7. The rheological properties of O/W emulsions stabilized by ChiNCs were compared to those stabilized by synthetic surfactants, namely Tween 80 and CTAB. The emulsions stabilized by ChiNCs demonstrated higher yield stress and lower shear viscosity compared to those stabilized by synthetic surfactants. This indicates that ChiNC-stabilized emulsions are more stable and less prone to breakdown. Based on these findings, ChiNCs show promise as an alternative to synthetic surfactants for stabilizing O/W emulsions.

Keywords: chitin nanocrystals, colloidal pickering, emulsion rheology, oil-in-water, synthetic surfactant

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14925 Role of Estrogen Receptor-alpha in Mammary Carcinoma by Single Nucleotide Polymorphisms and Molecular Docking: An In-silico Analysis

Authors: Asif Bilal, Fouzia Tanvir, Sibtain Ahmad

Abstract:

Estrogen receptor alpha, also known as estrogen receptor-1, is highly involved in risk of mammary carcinoma. The objectives of this study were to identify non-synonymous SNPs of estrogen receptor and their association with breast cancer and to identify the chemotherapeutic responses of phytochemicals against it via in-silico study design. For this purpose, different online tools. to identify pathogenic SNPs the tools were SIFT, Polyphen, Polyphen-2, fuNTRp, SNAP2, for finding disease associated SNPs the tools SNP&GO, PhD-SNP, PredictSNP, MAPP, SNAP, MetaSNP, PANTHER, and to check protein stability Mu-Pro, I-Mutant, and CONSURF were used. Post-translational modifications (PTMs) were detected by Musitedeep, Protein secondary structure by SOPMA, protein to protein interaction by STRING, molecular docking by PyRx. Seven SNPs having rsIDs (rs760766066, rs779180038, rs956399300, rs773683317, rs397509428, rs755020320, and rs1131692059) showing mutations on I229T, R243C, Y246H, P336R, Q375H, R394S, and R394H, respectively found to be completely deleterious. The PTMs found were 96 times Glycosylation; 30 times Ubiquitination, a single time Acetylation; and no Hydroxylation and Phosphorylation were found. The protein secondary structure consisted of Alpha helix (Hh) is (28%), Extended strand (Ee) is (21%), Beta turn (Tt) is 7.89% and Random coil (Cc) is (44.11%). Protein-protein interaction analysis revealed that it has strong interaction with Myeloperoxidase, Xanthine dehydrogenase, carboxylesterase 1, Glutathione S-transferase Mu 1, and with estrogen receptors. For molecular docking we used Asiaticoside, Ilekudinuside, Robustoflavone, Irinoticane, Withanolides, and 9-amin0-5 as ligands that extract from phytochemicals and docked with this protein. We found that there was great interaction (from -8.6 to -9.7) of these ligands of phytochemicals at ESR1 wild and two mutants (I229T and R394S). It is concluded that these SNPs found in ESR1 are involved in breast cancer and given phytochemicals are highly helpful against breast cancer as chemotherapeutic agents. Further in vitro and in vivo analysis should be performed to conduct these interactions.

Keywords: breast cancer, ESR1, phytochemicals, molecular docking

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14924 Dissociation of Hydrophobic Interactions in Whey Protein Polymers: Molecular Characterization Using Dilute Solution Viscometry

Authors: Ahmed S. Eissa

Abstract:

Whey represents about 85-95% of the milk volume and about 55% of milk nutrients. Whey proteins are of special importance in formulated foods due to their rich nutritional and functional benefits. Whey proteins form large polymers upon heating to a temperature greater than the denaturation temperature. Hydrophobic interactions play an important role in building whey protein polymers. In this study, dissociation of hydrophobic interactions of whey protein polymers was done by adding Sodium Dodecyl Sulphonate (SDS). At low SDS concentrations, protein polymers were dissociated to smaller chains, as revealed by dilution solution viscometry (DSV). Interestingly, at higher SDS concentrations, polymer molecules got larger in size. Intrinsic viscosity was increased to many folds when raising the SDS concentration from 0.5% to 2%. Complex molecular arrangement leads to the formation of larger macromolecules, due to micelle formation. The study opens a venue for manipulating and enhancing whey protein functional properties by manipulating the hydrophobic interactions.

Keywords: whey proteins, hydrophobic interactions, SDS

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14923 Removal of Protein from Chromium Tanning Bath by Biological Treatment Using Pseudomonas sp.

Authors: Amel Benhadji, Mourad Taleb Ahmed, Rachida Maachi

Abstract:

The challenge for the new millennium is to develop an industrial system that has minimal socio-ecological impacts, without compromising quality of life. Leather industry is one of these industries demanding environmentally friendly products. In this study, we investigated the possibility of applying innovative low cost biological treatment using Pseudomonas aeruginosa. This strain tested the efficiency of the batch biological treatment in the recovery of protein and hexavalent chromium from chromium tanning bath. We have compared suspended and fixed bacteria culture. The results showed the removal of the total protein of treatment and a decrease of hexavalent chromium concentration is during the treatment. The better efficiency of the biological treatment is obtained when using fixed culture of P. aeruginosa.

Keywords: tanning wastewater, biological treatment, protein removal, hexavalent chromium

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14922 Recovery of Proteins from EDAM Whey Using Membrane Ultrafiltration

Authors: F. Yelles-Allam, A. A. Nouani

Abstract:

In Algeria, whey is discarded without any treatment and this causes not only pollution problem, but also a loss in nutritive components of milk. In this paper, characterization of EDAM whey, which is resulted from pasteurised mixture of cow’s milk and skim milk, and recovery of whey protein by ultrafiltration / diafiltration, was studied. The physical-chemical analysis of whey has emphasized on its pollutant and nutritive characteristics. In fact, its DBO5 and DCO are 49.33, and 127.71 gr of O2/l of whey respectively. It contains: fat (1,90±0,1 gr/l), lactose (47.32±1,57 gr/l), proteins (8.04±0,2 gr/l) and ashes (5,20±0,15 gr/l), calcium (0,48±0,04 gr/l), Na (1.104gr/l), K (1.014 gr/l), Mg (0.118 gr/l) and P (0.482 gr/l). Ultrafiltration was carried out in a polyetersulfone membrane with a cut-off of 10K. Its hydraulic intrinsic resistance and permeability are respectively: 2.041.1012 m-1 and 176,32 l/h.m2 at PTM of 1 bar. The retentate obtained at FC6, contains 16,33g/l of proteins and 70,25 g/l of dry matter. The retention rate of protein is 97, 7% and the decrease in DBO5 and DCO are at 18.875 g /l and 42.818 g/l respectively. Diafiltration performed on protein concentrates allowed the complete removal of lactose and minerals. The ultrafiltration of the whey before the disposal is an alternative for Algéria dairy industry.

Keywords: diafiltration, DBO, DCO, protein, ultrafiltration, whey

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14921 Polarimetric Synthetic Aperture Radar Data Classification Using Support Vector Machine and Mahalanobis Distance

Authors: Najoua El Hajjaji El Idrissi, Necip Gokhan Kasapoglu

Abstract:

Polarimetric Synthetic Aperture Radar-based imaging is a powerful technique used for earth observation and classification of surfaces. Forest evolution has been one of the vital areas of attention for the remote sensing experts. The information about forest areas can be achieved by remote sensing, whether by using active radars or optical instruments. However, due to several weather constraints, such as cloud cover, limited information can be recovered using optical data and for that reason, Polarimetric Synthetic Aperture Radar (PolSAR) is used as a powerful tool for forestry inventory. In this [14paper, we applied support vector machine (SVM) and Mahalanobis distance to the fully polarimetric AIRSAR P, L, C-bands data from the Nezer forest areas, the classification is based in the separation of different tree ages. The classification results were evaluated and the results show that the SVM performs better than the Mahalanobis distance and SVM achieves approximately 75% accuracy. This result proves that SVM classification can be used as a useful method to evaluate fully polarimetric SAR data with sufficient value of accuracy.

Keywords: classification, synthetic aperture radar, SAR polarimetry, support vector machine, mahalanobis distance

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14920 Isolation and Characterization White Spot Syndrome Protein Envelope Protein 19 from Black Tiger Shrimp (Penaeus monodon)

Authors: Andi Aliah Hidayani, Asmi Citra Malina A. R. Tassakka, Andi Parenrengi

Abstract:

Vanname Shrimp is one of the high yielding varieties that are more resistant to virus attacks. However, now this shrimp more death due to virus attack such as white spot disease caused by white spot syndrome virus (WSSV). Various efforts have done to prevent the disease, like immunostimulatory, probiotics, and vaccine. White spot syndrome virus (WSSV) envelope protein VP19 gene is important because of its involvement in the system infection of shrimp. This study aimed to isolate and characterize an envelope protein VP19 – encoding gene of WSSV using WSSV infected Vanname Shrimp sample from some areas in South Sulawesi (Pangkep, Barru and Pinrang). The genomic of DNA were isolated from shrimp muscle using DTAB-CTAB method. Isolation of gene encoding envelope protein VP19 WSSV ws successfully performed with the results of the length of DNA fragment was 387 bp. The results of homology analysis using BLASTn homology suggested that these isolates genes from Barru, Pangkep and Pinrang have closest relationship with isolates from Mexican.

Keywords: vanname, shrimp, WSSV, viral protein 19

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14919 EverPro as the Missing Piece in the Plant Protein Portfolio to Aid the Transformation to Sustainable Food Systems

Authors: Aylin W Sahin, Alice Jaeger, Laura Nyhan, Gregory Belt, Steffen Münch, Elke K. Arendt

Abstract:

Our current food systems cause an increase in malnutrition resulting in more people being overweight or obese in the Western World. Additionally, our natural resources are under enormous pressure and the greenhouse gas emission increases yearly with a significant contribution to climate change. Hence, transforming our food systems is of highest priority. Plant-based food products have a lower environmental impact compared to their animal-based counterpart, representing a more sustainable protein source. However, most plant-based protein ingredients, such as soy and pea, are lacking indispensable amino acids and extremely limited in their functionality and, thus, in their food application potential. They are known to have a low solubility in water and change their properties during processing. The low solubility displays the biggest challenge in the development of milk alternatives leading to inferior protein content and protein quality in dairy alternatives on the market. Moreover, plant-based protein ingredients often possess an off-flavour, which makes them less attractive to consumers. EverPro, a plant-protein isolate originated from Brewer’s Spent Grain, the most abundant by-product in the brewing industry, represents the missing piece in the plant protein portfolio. With a protein content of >85%, it is of high nutritional value, including all indispensable amino acids which allows closing the protein quality gap of plant proteins. Moreover, it possesses high techno-functional properties. It is fully soluble in water (101.7 ± 2.9%), has a high fat absorption capacity (182.4 ± 1.9%), and a foaming capacity which is superior to soy protein or pea protein. This makes EverPro suitable for a vast range of food applications. Furthermore, it does not cause changes in viscosity during heating and cooling of dispersions, such as beverages. Besides its outstanding nutritional and functional characteristics, the production of EverPro has a much lower environmental impact compared to dairy or other plant protein ingredients. Life cycle assessment analysis showed that EverPro has the lowest impact on global warming compared to soy protein isolate, pea protein isolate, whey protein isolate, and egg white powder. It also contributes significantly less to freshwater eutrophication, marine eutrophication and land use compared the protein sources mentioned above. EverPro is the prime example of sustainable ingredients, and the type of plant protein the food industry was waiting for: nutritious, multi-functional, and environmentally friendly.

Keywords: plant-based protein, upcycled, brewers' spent grain, low environmental impact, highly functional ingredient

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14918 Expression of Tissue Plasminogen Activator in Transgenic Tobacco Plants by Signal Peptides Targeting for Delivery to Apoplast, Endoplasmic Reticulum and Cytosol Spaces

Authors: Sadegh Lotfieblisofla, Arash Khodabakhshi

Abstract:

Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.

Keywords: tPA, recombinant, transgenic, tobacco

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14917 Designing Next Generation Platforms for Recombinant Protein Production by Genome Engineering of Escherichia coli

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

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14916 Role of Mismatch Repair Protein Expression in Colorectal Cancer: A Study from North India

Authors: Alka Yadav, Mayank Jain, Rajan Saxena, Niraj Kumari, Narendra Krishnani, Ashok Kumar

Abstract:

Purpose: To study the mismatch repair (MMR) protein expression and its clinicopathological correlation in colorectal cancer patients in North India. Methods: A prospective study was conducted on histologically proven 52 (38 males and 14 females) patients with adenocarcinoma of colorectum. MMR protein loss was determined by using immunohistochemistry for MLH1, MSH2, PMS2 and MSH6. Results: 52 patients (38 males and 14 females) underwent resection for colorectal cancer with the median age of 52 years (16-81 years). 35% of the patients (n=18) were younger than 50 years of the age. 3 patients had associated history of malignancy in the family. 29 (56%) patients had right colon cancer, 9 (17%) left colon cancer and 14 (27%) rectal cancer. 2 patients each had synchronous and metachronous cancer. Histology revealed well-differentiated tumour in 16, moderately differentiated in 10 and poorly differentiated tumour in 26 patients. MMR protein loss was seen in 15 (29%) patients. Seven (46%) of these patients were less than 50 years of age. Combined loss of MSH2 and MSH6 was seen most commonly and it was found in 6 patients. 12 (80%) patients with MMR protein loss had tumour located proximal to the splenic flexure compared to 3 (20%) located distal to the splenic flexure. There was no difference in MMR protein loss based on patients' age, gender, degree of tumour differentiation, stage of the disease and tumour histological characteristics. Conclusions: This study revealed that there was less than 30% MMR protein loss in colorectal cancer patients. The loss was most commonly seen in right sided colon cancer than left. A larger study is further required to validate these findings.

Keywords: colorectal cancer, mismatch repair protein, immunohitochemistry, clinicopathological correlation

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14915 Targetting T6SS of Klebsiella pneumoniae for Assessment of Immune Response in Mice for Therapeutic Lead Development

Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

Abstract:

Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

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14914 Analytical Modeling of Globular Protein-Ferritin in α-Helical Conformation: A White Noise Functional Approach

Authors: Vernie C. Convicto, Henry P. Aringa, Wilson I. Barredo

Abstract:

This study presents a conformational model of the helical structures of globular protein particularly ferritin in the framework of white noise path integral formulation by using Associated Legendre functions, Bessel and convolution of Bessel and trigonometric functions as modulating functions. The model incorporates chirality features of proteins and their helix-turn-helix sequence structural motif.

Keywords: globular protein, modulating function, white noise, winding probability

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14913 Targeting Mre11 Nuclease Overcomes Platinum Resistance and Induces Synthetic Lethality in Platinum Sensitive XRCC1 Deficient Epithelial Ovarian Cancers

Authors: Adel Alblihy, Reem Ali, Mashael Algethami, Ahmed Shoqafi, Michael S. Toss, Juliette Brownlie, Natalie J. Tatum, Ian Hickson, Paloma Ordonez Moran, Anna Grabowska, Jennie N. Jeyapalan, Nigel P. Mongan, Emad A. Rakha, Srinivasan Madhusudan

Abstract:

Platinum resistance is a clinical challenge in ovarian cancer. Platinating agents induce DNA damage which activate Mre11 nuclease directed DNA damage signalling and response (DDR). Upregulation of DDR may promote chemotherapy resistance. Here we have comprehensively evaluated Mre11 in epithelial ovarian cancers. In clinical cohort that received platinum- based chemotherapy (n=331), Mre11 protein overexpression was associated with aggressive phenotype and poor progression free survival (PFS) (p=0.002). In the ovarian cancer genome atlas (TCGA) cohort (n=498), Mre11 gene amplification was observed in a subset of serous tumours (5%) which correlated highly with Mre11 mRNA levels (p<0.0001). Altered Mre11 levels was linked with genome wide alterations that can influence platinum sensitivity. At the transcriptomic level (n=1259), Mre11 overexpression was associated with poor PFS (p=0.003). ROC analysis showed an area under the curve (AUC) of 0.642 for response to platinum-based chemotherapy. Pre-clinically, Mre11 depletion by gene knock down or blockade by small molecule inhibitor (Mirin) reversed platinum resistance in ovarian cancer cells and in 3D spheroid models. Importantly, Mre11 inhibition was synthetically lethal in platinum sensitive XRCC1 deficient ovarian cancer cells and 3D-spheroids. Selective cytotoxicity was associated with DNA double strand break (DSB) accumulation, S-phase cell cycle arrest and increased apoptosis. We conclude that pharmaceutical development of Mre11 inhibitors is a viable clinical strategy for platinum sensitization and synthetic lethality in ovarian cancer.

Keywords: MRE11; XRCC1, ovarian cancer, platinum sensitization, synthetic lethality

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14912 Effect of Synthetic L-Lysine and DL-Methionine Amino Acids on Performance of Broiler Chickens

Authors: S. M. Ali, S. I. Mohamed

Abstract:

Reduction of feed cost for broiler production is at most importance in decreasing the cost of production. The objectives of this study were to evaluate the use of synthetic amino acids (L-lysine – DL-methionine) instead of super concentrate and groundnut cake versus meat powder as protein sources. A total of 180 male broiler chicks (Cobb – strain) at 15 day of age (DOA) were selected according to their average body weight (380 g) from a broiler chicks flock at Elbashair Farm. The chicks were randomly divided into six groups of 30 chicks. Each group was further sub divided into three replicates with 10 birds. Six experimental diets were formulated. The first diet contained groundnut cake and super concentrate as the control (GNC + C); in the second diet, meat powder and super concentrate (MP + C) were used. The third diet contained groundnut cake and amino acids (GNC + AA); the forth diet contained meat powder and amino acids (MP + AA). The fifth diet contained groundnut cake, meat powder and super concentrate (GNC + MP + C) and the sixth diet contained groundnut cake, meat powder and amino acids (GNC + MP + AA). The formulated rations were randomly assigned for the different sub groups in a completely randomized design of six treatments and three replicates. Weekly feed intake, body weight and mortality were recorded and body weight gain and feed conversion ratio were calculated. At the end of the experiment (49 DOA), nine birds from each treatment were slaughtered. Live body weight, carcass weight, head, shank, and some internal organs (gizzard, heart, liver, small intestine, and abdominal fat pad) weights were taken. For the overall experimental period the (GNC + C +MP) consumed significantly (P≤0.01) the highest cumulative feed while the (MP + AA) group consumed the lowest amount of feed. The (GNC + C) and the (GNC + AA) groups had the heaviest live body weight while (MP + AA) had the lowest live body weight. The overall FCR was significantly (P≤0.01) the best for (GNC + AA) group while the (MP + AA) reported the worst FCR. However, the (GNC + AA) had significantly (P≤0.01) the lowest AFP. The (GNC + MP + Con) group had the highest dressing % while the (MP + AA) group had the lowest dressing %. It is concluded that amino acids can be used instead of super concentrate in broiler feeding with perfect performance and less cost and that meat powder is not advisable to be used with amino acids.

Keywords: broiler chickens, DL-lysine, methionine, performance

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14911 Use RP-HPLC To Investigate Factors Influencing Sorghum Protein Extraction

Authors: Khaled Khaladi, Rafika Bibi, Hind Mokrane, Boubekeur Nadjemi

Abstract:

Sorghum (Sorghum bicolor (L.) Moench) is an important cereal crop grown in the semi-arid tropics of Africa and Asia due to its drought tolerance. Sorghum grain has protein content varying from 6 to 18%, with an average of 11%, Sorghum proteins can be broadly classified into prolamin and non-prolamin proteins. Kafirins, the major storage proteins, are classified as prolamins, and as such, they contain high levels of proline and glutamine and are soluble in non-polar solvents such as aqueous alcohols. Kafirins account for 77 to 82% of the protein in the endosperm, whereas non-prolamin proteins (namely, albumins, globulins, and glutelins) make up about 30% of the proteins. To optimize the extraction of sorghum proteins, several variables were examined: detergent type and concentration, reducing agent type and concentration, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC.

Keywords: sorghum, protein extraction, detergent, food science

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14910 Quantifying the Protein-Protein Interaction between the Ion-Channel-Forming Colicin A and the Tol Proteins by Potassium Efflux in E. coli Cells

Authors: Fadilah Aleanizy

Abstract:

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

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

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14909 Delivery of Positively Charged Proteins Using Hyaluronic Acid Microgels

Authors: Elaheh Jooybar, Mohammad J. Abdekhodaie, Marcel Karperien, Pieter J. Dijkstra

Abstract:

In this study, hyaluronic acid (HA) microgels were developed for the goal of protein delivery. First, a hyaluronic acid-tyramine conjugate (HA-TA) was synthesized with a degree of substitution of 13 TA moieties per 100 disaccharide units. Then, HA-TA microdroplets were produced using a water in oil emulsion method and crosslinked in the presence of horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). Loading capacity and the release kinetics of lysozyme and BSA, as model proteins, were investigated. It was shown that lysozyme, a cationic protein, can be incorporated efficiently in the HA microgels, while the loading efficiency for BSA, as a negatively charged protein, is low. The release profile of lysozyme showed a sustained release over a period of one month. The results demonstrated that the HA-TA microgels are a good carrier for spatial delivery of cationic proteins for biomedical applications.

Keywords: microgel, inverse emulsion, protein delivery, hyaluronic acid, crosslinking

Procedia PDF Downloads 148
14908 Invasive Asian Carp Fish Species: A Natural and Sustainable Source of Methionine for Organic Poultry Production

Authors: Komala Arsi, Ann M. Donoghue, Dan J. Donoghue

Abstract:

Methionine is an essential dietary amino acid necessary to promote growth and health of poultry. Synthetic methionine is commonly used as a supplement in conventional poultry diets and is temporarily allowed in organic poultry feed for lack of natural and organically approved sources of methionine. It has been a challenge to find a natural, sustainable and cost-effective source for methionine which reiterates the pressing need to explore potential alternatives of methionine for organic poultry production. Fish have high concentrations of methionine, but wild-caught fish are expensive and adversely impact wild fish populations. Asian carp (AC) is an invasive species and its utilization has the potential to be used as a natural methionine source. However, to our best knowledge, there is no proven technology to utilize this fish as a methionine source. In this study, we co-extruded Asian carp and soybean meal to form a dry-extruded, methionine-rich AC meal. In order to formulate rations with the novel extruded carp meal, the product was tested on cecectomized roosters for its amino acid digestibility and total metabolizable energy (TMEn). Excreta was collected and the gross energy, protein content of the feces was determined to calculate Total Metabolizable Energy (TME). The methionine content, digestibility and TME values were greater for the extruded AC meal than control diets. Carp meal was subsequently tested as a methionine source in feeds formulated for broilers, and production performance (body weight gain and feed conversion ratio) was assessed in comparison with broilers fed standard commercial diets supplemented with synthetic methionine. In this study, broiler chickens were fed either a control diet with synthetic methionine or a treatment diet with extruded AC meal (8 replicates/treatment; n=30 birds/replicate) from day 1 to 42 days of age. At the end of the trial, data for body weights, feed intake and feed conversion ratio (FCR) was analyzed using one-way ANOVA with Fisher LSD test for multiple comparisons. Results revealed that birds on AC diet had body weight gains and feed intake comparable to diets containing synthetic methionine (P > 0.05). Results from the study suggest that invasive AC-derived fish meal could potentially be an effective and inexpensive source of sustainable natural methionine for organic poultry farmers.

Keywords: Asian carp, methionine, organic, poultry

Procedia PDF Downloads 134
14907 Bioinformatics and Molecular Biological Characterization of a Hypothetical Protein SAV1226 as a Potential Drug Target for Methicillin/Vancomycin-Staphylococcus aureus Infections

Authors: Nichole Haag, Kimberly Velk, Tyler McCune, Chun Wu

Abstract:

Methicillin/multiple-resistant Staphylococcus aureus (MRSA) are infectious bacteria that are resistant to common antibiotics. A previous in silico study in our group has identified a hypothetical protein SAV1226 as one of the potential drug targets. In this study, we reported the bioinformatics characterization, as well as cloning, expression, purification and kinetic assays of hypothetical protein SAV1226 from methicillin/vancomycin-resistant Staphylococcus aureus Mu50 strain. MALDI-TOF/MS analysis revealed a low degree of structural similarity with known proteins. Kinetic assays demonstrated that hypothetical protein SAV1226 is neither a domain of an ATP dependent dihydroxyacetone kinase nor of a phosphotransferase system (PTS) dihydroxyacetone kinase, suggesting that the function of hypothetical protein SAV1226 might be misannotated on public databases such as UniProt and InterProScan 5.

Keywords: Methicillin-resistant Staphylococcus aureus, dihydroxyacetone kinase, essential genes, drug target, phosphoryl group donor

Procedia PDF Downloads 383
14906 Nutritional Potential and Functionality of Whey Powder Influenced by Different Processing Temperature and Storage

Authors: Zarmina Gillani, Nuzhat Huma, Aysha Sameen, Mulazim Hussain Bukhari

Abstract:

Whey is an excellent food ingredient owing to its high nutritive value and its functional properties. However, composition of whey varies depending on composition of milk, processing conditions, processing method, and its whey protein content. The aim of this study was to prepare a whey powder from raw whey and to determine the influence of different processing temperatures (160 and 180 °C) on the physicochemical, functional properties during storage of 180 days and on whey protein denaturation. Results have shown that temperature significantly (P < 0.05) affects the pH, acidity, non-protein nitrogen (NPN), protein total soluble solids, fat and lactose contents. Significantly (p < 0.05) higher foaming capacity (FC), foam stability (FS), whey protein nitrogen index (WPNI), and a lower turbidity and solubility index (SI) were observed in whey powder processed at 160 °C compared to whey powder processed at 180 °C. During storage of 180 days, slow but progressive changes were noticed on the physicochemical and functional properties of whey powder. Reverse phase-HPLC analysis revealed a significant (P < 0.05) effect of temperature on whey protein contents. Denaturation of β-Lactoglobulin is followed by α-lacalbumin, casein glycomacropeptide (CMP/GMP), and bovine serum albumin (BSA).

Keywords: whey powder, temperature, denaturation, reverse phase, HPLC

Procedia PDF Downloads 275
14905 Towards the Inhibition Mechanism of Lysozyme Fibrillation by Hydrogen Sulfide

Authors: Indra Gonzalez Ojeda, Tatiana Quinones, Manuel Rosario, Igor Lednev, Juan Lopez Garriga

Abstract:

Amyloid fibrils are stable aggregates of misfolded protein associated with many neurodegenerative disorders. It has been shown that hydrogen sulfide (H2S), inhibits the fibrillation of lysozyme through the formation of trisulfide (S-S-S) bonds. However, the overall mechanism remains elusive. Here, the concentration dependence of H2S effect was investigated using Atomic force microscopy (AFM), non-resonance Raman spectroscopy, Deep-UV Raman spectroscopy and circular dichroism (CD). It was found that small spherical aggregates with trisulfide bonds and a unique secondary structure were formed instead of amyloid fibrils when adding concentrations of 25 mM and 50 mM of H2S. This could indicate that H2S might serve as a protecting agent for the protein. However, further characterization of these aggregates and their trisulfide bonds is needed to fully unravel the function H2S has on protein fibrillation.

Keywords: amyloid fibrils, hydrogen sulfide, protein folding, raman spectroscopy

Procedia PDF Downloads 199
14904 Protein Remote Homology Detection by Using Profile-Based Matrix Transformation Approaches

Authors: Bin Liu

Abstract:

As one of the most important tasks in protein sequence analysis, protein remote homology detection has been studied for decades. Currently, the profile-based methods show state-of-the-art performance. Position-Specific Frequency Matrix (PSFM) is widely used profile. However, there exists noise information in the profiles introduced by the amino acids with low frequencies. In this study, we propose a method to remove the noise information in the PSFM by removing the amino acids with low frequencies called Top frequency profile (TFP). Three new matrix transformation methods, including Autocross covariance (ACC) transformation, Tri-gram, and K-separated bigram (KSB), are performed on these profiles to convert them into fixed length feature vectors. Combined with Support Vector Machines (SVMs), the predictors are constructed. Evaluated on two benchmark datasets, and experimental results show that these proposed methods outperform other state-of-the-art predictors.

Keywords: protein remote homology detection, protein fold recognition, top frequency profile, support vector machines

Procedia PDF Downloads 104
14903 C-Reactive Protein in Patients with Type 2 Diabetes Mellitus

Authors: Athar Hussain Memon

Abstract:

Objectives: We tried to determine the frequency of raised C-reactive protein (CRP) in patients with type 2 diabetes mellitus. Patients and Methods: This cross-sectional descriptive study of six months study was conducted at Liaquat University Hospital Hyderabad from March 2013 to August 2013. All diabetic patients of ≥35 years age of either gender for >01 year duration visited at OPD were evaluated for C-reactive protein and their glycemic status by hemoglobin A1c. The data was analyzed in SPSS and the frequency and percentage were calculated. Results: During six month study period, total 100 diabetic patients were evaluated for C-reactive protein. The majority of patients were from urban areas 75/100 (75%). The mean ±SD for age of patients with diabetes mellitus was 51.63±7.82. The mean age ±SD of patient with raised CRP was 53±7.21. The mean ±SD for HbA1c in patients with raised CRP is 9.55±1.73. The mean random blood sugar level in patients with raised CRP was 247.42 ± 6.62. The majority of subjects were of 50-69 years of age group with female predominance (p=0.01) while the CRP was raised in 70 (70%) patients in relation to age (p=0.02) and gender (p=0.01), respectively. Both HbA1c and CRP were raised in 64.9% (p=0.04) in patients with type 2 diabetes mellitus. The mean ±SD of CRP was 5.8±1.21 while for male and female individuals with raised CRP was 3.52±1.22 and 5.7±1.63, respectively. Conclusions: The raised CRP was observed in patients with type 2 diabetes mellitus.

Keywords: diabetes mellitus, C-reactive protein, hemoglobin A1c, diabetes and metabolism

Procedia PDF Downloads 390
14902 Production of Recombinant VP2 Protein of Canine Parvovirus Type 2c Using Baculovirus Expression System

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

Abstract:

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

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

Procedia PDF Downloads 123
14901 The Performance of Six Exotic Perennial Grass Species in the Central Region of Saudi Arabia

Authors: A. Alsoqeer

Abstract:

The establishment, dry matter production and feeding value of six perennial grasses were measured over two growing seasons in a field experiments. The experiments were conducted at the Agricultural and Veterinary Medicine Research Station, Faculty of Agriculture and Veterinary Medicine, Qassim University, Kingdom of Saudi Arabia in 2009 and 2010 seasons. The six perennial grasses were: creeping bluegrass (Bothriochloa insculpta cv. Bisset), digit grass (Digitaria smutsi), Jarra digit grass (Digitaria milanjiana), panic (Panicum coloratum cv. Bambatsii), Sabi grass (Urochloa mosambicensis) and setaria (Setaria sphacelata cv. Kazungula). The experimental design used was a completely randomized block design with four replications. The results revealed significant differences among plant species of all agronomic characters and quality traits in the first year, while in the second year, plant species differed significantly for quality traits only. D. smutsi had a superior performance for all agronomic characters, however, it had the lowest values in protein content in the two years comparing with other genotypes. D. milanjiana and U. mosambicensis showed high values in dry matter yield and protein content in the first year, but showed a very poor performance in the second year because most of plants were die due to the low temperatures in the winter. These two species appear to be suitable for annual cultivation. The other species tolerate the cold winter and were a highly productive in the second year.

Keywords: dry mater yield, grass species, cuts, quality traits, crude protein content

Procedia PDF Downloads 293
14900 Phase Transition in Iron Storage Protein Ferritin

Authors: Navneet Kaur, S. D. Tiwari

Abstract:

Ferritin is a protein which present in the blood of mammals. It maintains the need of iron inside the body. It has an antiferromagnetic iron core, 7-8 nm in size, which is encapsulated inside a protein cage. The thickness of this protein shell is about 2-3 nm. This protein shell reduces the interaction among particles and make ferritin a model superparamagnet. The major composition of ferritin core is mineral ferrihydrite. The molecular formula of ferritin core is (FeOOH)8[FeOOPO3H2]. In this study, we discuss the phase transition of ferritin. We characterized ferritin using x-ray diffractometer, transmission electron micrograph, thermogravimetric analyzer and vibrating sample magnetometer. It is found that ferritin core is amorphous in nature with average particle size of 8 nm. The thermogravimetric and differential thermogravimetric analysis curves shows mass loss at different temperatures. We heated ferritin at these temperatures. It is found that ferritin core starts decomposing after 390^o C. At 1020^o C, the ferritin core is finally converted to alpha phase of iron oxide. Magnetization behavior of final sample clearly shows the iron oxyhydroxide core is completely converted to alpha iron oxide.

Keywords: Antiferromagnetic, Ferritin, Phase, Superparamagnetic

Procedia PDF Downloads 93
14899 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

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