Search results for: Soluble protein

Authors: Le Thuy Mai, Vu Nguyen Thanh

Abstract:

β-Glucosidase is an important enzyme for production of ethanol from lignocellulose. With hydrolytic activity on cellooligosaccharides, especially cellobiose, β-glucosidase removes product inhibitory effect on cellulases and forms fermentable sugars. In this study, β-glucosidase encoding gene (BGL1) from traditional starter yeast Saccharomycosis fibuligera BMQ908 was cloned and expressed in Pichia pastoris. BGL1 of S. fibuligera BMQ 908 shared 98% nucleotide homology with the closest GenBank sequence (M22475) but identity in amino-acid sequences of catalytic domains. Recombinant plasmid pPICZαA/BGL1 containing the sequence encoding BGL1 mature protein and α-factor secretion signal was constructed and transformed into methylotrophic yeast P. pastoris by electroporation. The recombinant strain produced single extracellular protein with molecular weight of 120 kDa and cellobiase activity of 60 IU/ml. The optimum pH of the recombinant β-glucosidase was 5.0 and the optimum temperature was 50°C.

Keywords: β-Glucosidase, Pichia pastoris, Saccharomycopsisfibuligera, recombinant enzyme.

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Authors: Laima Silina, Ilze Gramatina, Lija Dukalska, Liga Skudra, Tatjana Rakcejeva, Dace Klava, Anita Blija

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The objective of the present study was to determine quality parameters changes of red wine marinade marinated venison during storage. Beef as a control was analysed. Protein, fat, moisture and pH content dynamics as well microbiological quality was analyzed. The meat pieces were marinated in red wine marinade at 4±2ºC temperature for 48±1h. Marinated meat was placed in polypropylene trays, hermetically sealed with high barrier polymer film Multibarrier 60 under modified atmosphere (CO₂ 40%+N₂ 60%) without and with oxygen absorber sachets, as a control packaging in air ambiance packed marinated venison and beef was used. Meat samples were analyzed after 0, 4, 7, 11 and 14 days of storage. During the storage of meat, protein and moisture content significantly (p<0.05) decreased, pH and colony forming units significantly (p<0.05) increased, fat content does not change in all treatments irrespective of the packaging method.

Keywords: Marinating, modified atmosphere, quality, venison.

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Authors: A. Radzevičius, P. Viškelis, J. Viškelis, R. Bobinaitė, R. Karklelienė, D. Juškevičienė

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Two cultivars ('Rutuliai', 'Saint Perrie') and five hybrids ('Tolstoi', 'Brooklyn', 'Tocayo', 'Benito', 'Tourist') of edible tomato (Lycopersicon esculentum Mill.) were investigated at the LRCAF Institute of Horticulture. The following fruit quality parameters were evaluated: the amount of lycopene, β-carotene, ascorbic acid, total and inverted sugar, sucrose, dry matter soluble solids in fresh tomato matter, also were determined fruit skin and flesh firmness, color indexes (CIE L*a*b*) and calculated hue angle (h°) with chroma (C).

Keywords: Carotenoids, cultivar, nutrition, tomato.

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Authors: Ngo Thanh Xuan, Mai Thi Hang, Vu Nguyen Thanh

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A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.

Keywords: Aspergillus niger XP, cloning, over expression, Pichia pastoris, phyA , phytase.

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Authors: O. Aluko, M. R. Brai, A. O. Adelore

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Healthy snack (cookie) was produced from corn flour and moringa seed flour blends. The samples were mixed in various proportions and analysed for proximate composition and functional characteristics. The healthy snack (cookies) was evaluated for sensory parameters of Colour, Crispness, Taste, Aroma and Overall Acceptability. The proximate analysis of the flour obtained from different proportion showed that proximate composition increased with increase in substitution level of moringa seed flour especially with protein, fat and crude fibre. The protein contents of samples range from 1.75 to 6.58, fat from 0.60 to 6.80, while fibre from 0.85 to 2.06. There was no significance difference in the functional properties of the blend when compared with 100% corn flour. Sensory evaluation results shows a significant difference in Colour, Taste, Crispness, Aroma and Overall Acceptability of healthy snack (cookies) sample from different blends at 5% significance level.

Keywords: Healthy snack, moringa.

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Authors: Hong Ding, Fatiha Benslimane, Isra Marei, Chris R. Triggle

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Hyperglycaemia is a key factor that contributes to the development of diabetes-related microvascular disease and a major risk factor for endothelial dysfunction. In the current study, we have explored glucose-induced abnormal intracellular calcium (Ca2+ i) homeostasis in mouse microvessel endothelial cells (MMECs) in high glucose (HG) (40mmol/L) versus control (low glucose, LG) (11 mmol/L). We demonstrated that the exposure of MMECs to HG for 3 days did not change basal Ca2+ i, however, there was a significant increase of acetylcholine-induced Ca2+ entry. Western blots illustrated that exposure to HG also increased STIM1 (Stromal Interaction Molecule 1), but not Orai1 (the pore forming subunit), protein expression levels. Although the link between HG-induced changes in STIM1 expression, enhanced Ca2+ entry and endothelial dysfunction requires further study, the current data are suggestive that targeting these pathways may reduce the impact of HG on endothelial function.

Keywords: store-operated calcium entry, hyperglycaemia, STIM1, endothelial dysfunction

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Authors: A. E. O. Malau-Aduli, B. W. B. Holman, P. A. Lane

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Our objectives were to evaluate the effects of sire breed, type of protein supplement, level of supplementation and sex on wool spinning fineness (SF), its correlations with other wool characteristics and prediction accuracy in F1 Merino crossbred lambs. Texel, Coopworth, White Suffolk, East Friesian and Dorset rams were mated with 500 purebred Merino dams at a ratio of 1:100 in separate paddocks within a single management system. The F1 progeny were raised on ryegrass pasture until weaning, before forty lambs were randomly allocated to treatments in a 5 x 2 x 2 x 2 factorial experimental design representing 5 sire breeds, 2 supplementary feeds (canola or lupins), 2 levels of supplementation (1% or 2% of liveweight) and sex (wethers or ewes). Lambs were supplemented for six weeks after an initial three weeks of adjustment, wool sampled at the commencement and conclusion of the feeding trial and analyzed for SF, mean fibre diameter (FD), coefficient of variation (CV), standard deviation, comfort factor (CF), fibre curvature (CURV), and clean fleece yield. Data were analyzed using mixed linear model procedures with sire fitted as a random effect, and sire breed, sex, supplementary feed type, level of supplementation and their second-order interactions as fixed effects. Sire breed (P<0.001), sex (P<0.004), sire breed x level of supplementation (P<0.004), and sire breed x sex (P<0.019) interactions significantly influenced SF. SF ranged from 22.7 ± 0.2μm in White Suffolk-sired lambs to 25.1 ± 0.2μm in East Friesian crossbred lambs. Ewes had higher SF than wethers. There were significant (P<0.001) correlations between SF and FD (0.93), CV (0.40), CF (-0.94) and CURV (-0.12). Its strong relationship with other wool quality traits enabled accurate predictions explaining up to about 93% of the observed variation. The interactions between sire breed genetics and nutrition will have an impact on the choices that dual-purpose sheep producers make when selecting sire breeds and protein supplementary feed levels to achieve optimal wool spinning fineness at the farmgate level. This will facilitate selective breeding programs being able to better account for SF and its interactions with other wool characteristics.

Keywords: Merino crossbred sheep, protein supplementation, sire breed, wool quality, wool spinning fineness

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Authors: S. B. Fasoyiro, K. A. Arowora

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Pigeon pea (Cajanus cajan) blanched for 20min was dehulled and milled into flour. The flour was incorporated into dried whole fermented maize (Ogi) at five levels. The resultant products were analyzed for chemical and pasting properties. The fortified Ogi samples were also assessed for sensory attributes: appearance, color, flavor, mouth feel and overall acceptability. The protein content in the whole Ogi fortified samples was in the range of 11.2-16.6% and crude fibre 3.22-3.46%. Fortified whole Ogi with pigeon pea at 30%, 40% and 50% of inclusion with pigeon pea flour has higher protein, crude fibre and ash content. Varying range of pasting quality was recorded for the blends, pasting temperature for fortified Obi was in the range of 45.3-49.50C and peak time 5.05-5.210C. The sensory acceptability of the whole Ogi fortified blends prepared into gruel has higher acceptability for various qualities in comparison with the traditional Ogi gruel.

Keywords: Maize Ogi, pigeon pea, chemical, pasting, sensory properties.

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Authors: Razvan Bocu, Dr Sabin Tabirca

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The study of proteomics reached unexpected levels of interest, as a direct consequence of its discovered influence over some complex biological phenomena, such as problematic diseases like cancer. This paper presents the latest authors- achievements regarding the analysis of the networks of proteins (interactome networks), by computing more efficiently the betweenness centrality measure. The paper introduces the concept of betweenness centrality, and then describes how betweenness computation can help the interactome net- work analysis. Current sequential implementations for the between- ness computation do not perform satisfactory in terms of execution times. The paper-s main contribution is centered towards introducing a speedup technique for the betweenness computation, based on modified shortest path algorithms for sparse graphs. Three optimized generic algorithms for betweenness computation are described and implemented, and their performance tested against real biological data, which is part of the IntAct dataset.

Keywords: Betweenness centrality, interactome networks, protein-protein interactions, sub-communities, sparse networks, speedup tech-nique, IntAct.

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Authors: Razvan Bocu, Sabin Tabirca

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The study of proteomics reached unexpected levels of interest, as a direct consequence of its discovered influence over some complex biological phenomena, such as problematic diseases like cancer. This paper presents a new technique that allows for an accurate analysis of the human interactome network. It is basically a two-step analysis process that involves, at first, the detection of each protein-s absolute importance through the betweenness centrality computation. Then, the second step determines the functionallyrelated communities of proteins. For this purpose, we use a community detection technique that is based on the edge betweenness calculation. The new technique was thoroughly tested on real biological data and the results prove some interesting properties of those proteins that are involved in the carcinogenesis process. Apart from its experimental usefulness, the novel technique is also computationally effective in terms of execution times. Based on the analysis- results, some topological features of cancer mutated proteins are presented and a possible optimization solution for cancer drugs design is suggested.

Keywords: Betweenness centrality, interactome networks, proteinprotein interactions, protein communities, cancer.

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Authors: H. Kiani, S. Moradi, B. Soltani Soulgani, S. Mousavian

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Desalting/dehydration plants (DDP) are often installed in crude oil production units in order to remove water-soluble salts from an oil stream. In order to optimize this process, desalting unit should be modeled. In this research, artificial neural network is used to model efficiency of desalting unit as a function of input parameter. The result of this research shows that the mentioned model has good agreement with experimental data.

Keywords: Desalting unit, Crude oil, Neural Networks, Simulation, Recovery, Separation.

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Authors: Masaki Yamaguchi, Akira Ito, Noriaki Okamoto, Yoshinori Kawabe, Masamichi Kamihira

Abstract:

Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced heat-inducible transgene expression system in which a heat-shock protein (HSP) promoter and tetracycline-responsive transactivator were combined. When the transactivator plasmid containing the tetracycline-responsive transactivator gene was co-transfected with the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP promoter without the transactivator. In vitro evaluation of the therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of these cells, indicating that this strategy is promising for cancer gene therapy.

Keywords: Inducible gene expression, Gene therapy, Hyperthermia, Heat shock protein, Tetracycline transactivator.

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Authors: Gnanaprakash K, Madhusudhana Chetty C, Ramkanth S, Alagusundaram M, Tiruvengadarajan VS, Angala Parameswari S, Mohamed Saleem TS

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Carbon tetrachloride (CCl4) is a well-known hepatotoxin and exposure to this chemical is known to induce oxidative stress and causes liver injury by the formation of free radicals. Flacourtia indica commonly known as 'Baichi' has been reported as an effective remedy for the treatment of a variety of diseases. The objective of this study was to investigate the hepatoprotective activity of aqueous extract of leaves of Flacourtia indica against CCl4 induced hepatotoxicity. Animals were pretreated with the aqueous extract of Flacourtia indica (250 & 500 mg/kg body weight) for one week and then challenged with CCl4 (1.5 ml/kg bw) in olive oil (1:1, v/v) on 7th day. Serum marker enzymes (ALP, AST, ALT, Total Protein & Total Bilirubin) and TBARS level (Marker for oxidative stress) were estimated in all the study groups. Alteration in the levels of biochemical markers of hepatic damage like AST, ALT, ALP, Total Protein, Total Bilirubin and lipid peroxides (TBARS) were tested in both CCl4 treated and extract treated groups. CCl4 has enhanced the AST, ALT, ALP and the Lipid peroxides (TBARS) in liver. Treatment of aqueous extract of Flacourtia indica leaves (250 & 500 mg/kg) exhibited a significant protective effect by altering the serum levels of AST, ALT, ALP, Total Protein, Total Bilirubin and liver TBARS. These biochemical observations were supported by histopathological study of liver sections. From this preliminary study it has been concluded that the aqueous extract of the leaves of Flacourtia indica protects liver against oxidative damages and could be used as an effective protector against CCl4 induced hepatic damage. Our findings suggested that Flacourtia indica possessed good hepatoprotective activity

Keywords: Carbon Tetrachloride, Flacourtia indica, Hepatoprotective activity, Oxidative stress

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Authors: V. Viliene, V. Sasyte, A. Raceviciute-Stupeliene, R. Gruzauskas

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In horse nutrition, the most suitable cereal for their rations composition could be defined as oats and barley. Oats have high nutritive value because it provides more protein, fiber, iron and zinc than other whole grains, has good taste, and an activity of stimulating metabolic changes in the body. Another cereal – barley is very similar to oats as a feed except for some characteristics that affect how it is used; however, barley is lower in fiber than oats and is classified as a "heavy" feed. The value of oats and barley grain, first of all is dependent on its composition. Near-infrared spectroscopy (NIRS) has long been considered and used as a significant method in component and quality analysis and as an emerging technology for authenticity applications for cereal quality control. This paper presents the chemical and amino acid composition of different varieties of barley and oats, also digestible energy of different cereals for horses. Ten different spring barley (n = 5) and oats (n = 5) varieties, grown in one location in Lithuania, were assayed for their chemical composition (dry matter, crude protein, crude fat, crude ash, crude fiber, starch) and amino acids content, digestible amino acids and amino acids digestibility. Also, the grains digestible energy for horses was calculated. The oats and barley samples reflectance spectra were measured by means of NIRS using Foss-Tecator DS2500 equipment. The chemical components: fat, crude protein, starch and fiber differed statistically (P<0.05) between the oats and barley varieties. The highest total amino acid content between oats was determined in variety Flamingsprofi (4.56 g/kg) and the lowest – variety Circle (3.57 g/kg), and between barley - respectively in varieties Publican (3.50 g/kg) and Sebastian (3.11 g/kg). The different varieties of oats digestible amino acid content varied from 3.11 g/kg to 4.07 g/kg; barley different varieties varied from 2.59 g/kg to 2.94 g/kg. The average amino acids digestibility of oats varied from 74.4% (Liz) to 95.6% (Fen) and in barley - from 75.8 % (Tre) to 89.6% (Fen). The amount of digestible energy in the analyzed varieties of oats and barley was an average compound 13.74 MJ/kg DM and 14.85 MJ/kg DM, respectively. An analysis of the results showed that different varieties of oats compared with barley are preferable for horse nutrition according to the crude fat, crude fiber, ash and separate amino acids content, but the analyzed barley varieties dominated the higher amounts of crude protein, the digestible Liz amount and higher DE content, and thus, could be recommended for making feed formulation for horses combining oats and barley, taking into account the chemical composition of using cereal varieties.

Keywords: Barley, digestive energy, horses, nutritional value, oats.

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Authors: Abu Salim Mustafa

Abstract:

Rv3873 is a relatively large size protein (371 amino acids in length) and its gene is located in the immunodominant genomic region of difference (RD)1 that is present in the genome of Mycobacterium tuberculosis but deleted from the genomes of all the vaccine strains of Bacillus Calmette Guerin (BCG) and most other mycobacteria. However, when tested for cellular immune responses using peripheral blood mononuclear cells from tuberculosis patients and BCG-vaccinated healthy subjects, this protein was found to be a major stimulator of cell mediated immune responses in both groups of subjects. In order to further identify the sequence of immunodominant epitopes and explore their Human Leukocyte Antigen (HLA)-restriction for epitope recognition, 24 peptides (25-mers overlapping with the neighboring peptides by 10 residues) covering the sequence of Rv3873 were synthesized chemically using fluorenylmethyloxycarbonyl chemistry and tested in cell mediated immune responses. The results of these experiments helped in the identification of an immunodominant peptide P9 that was recognized by people expressing varying HLA-DR types. Furthermore, it was also predicted to be a promiscuous binder with multiple epitopes for binding to HLA-DR, HLA-DP and HLA-DQ alleles of HLA-class II molecules that present antigens to T helper cells, and to HLA-class I molecules that present antigens to T cytotoxic cells. In addition, the evaluation of peptide P9 using an immunogenicity predictor server yielded a high score (0.94), which indicated a greater probability of this peptide to elicit a protective cellular immune response. In conclusion, P9, a peptide with multiple epitopes and ability to bind several HLA class I and class II molecules for presentation to cells of the cellular immune response, may be useful as a peptide-based vaccine against tuberculosis.

Keywords: Mycobacterium tuberculosis, Rv3873, peptides, vaccine

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Authors: Ines Hamdi, Mohamed Ben Ahmed

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The understanding of the system level of biological behavior and phenomenon variously needs some elements such as gene sequence, protein structure, gene functions and metabolic pathways. Challenging problems are representing, learning and reasoning about these biochemical reactions, gene and protein structure, genotype and relation between the phenotype, and expression system on those interactions. The goal of our work is to understand the behaviors of the interactions networks and to model their evolution in time and in space. We propose in this study an ontological meta-model for the knowledge representation of the genetic regulatory networks. Ontology in artificial intelligence means the fundamental categories and relations that provide a framework for knowledge models. Domain ontology's are now commonly used to enable heterogeneous information resources, such as knowledge-based systems, to communicate with each other. The interest of our model is to represent the spatial, temporal and spatio-temporal knowledge. We validated our propositions in the genetic regulatory network of the Aarbidosis thaliana flower

Keywords: Ontological model, spatio-temporal modeling, Genetic Regulatory Networks (GRNs), knowledge representation.

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Authors: M. Z. Othman

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The aim of this study was to investigate the effectiveness of anaerobic digestion for the treatment of wool scouring wastes. The experiments design comprised three ratios of waste (W) to seed(S) (W:S) of 25:75, 50:50 and 75:25, corresponding to 1.9. 1.7 and 1.5g tCOD/g TS, respectively, with or without chemicals addition. NH4Cl was added to the reactors as a source for nitrogen to achieve C:N:P of 420:14:3. A cationic flocculent was added at 0.5 and 0.75% to enhance flocculation of sludge. The results showed that the reactors that received W:S at a ratio of 25:75 produced the largest volume of biogas. The final soluble COD (sCOD) was below the limits for discharge to the sewer system.

Keywords: Anaerobic digestion, wool processing waste, organicloading, biogas.

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Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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This study was conducted to determine that carrot powder obtain by different drying methods (oven and vacuum-freeze dryer) of carrot unfit for human consumption that whether feed additives in animal nutrition or not. Carrots randomly divided 2 groups. First group was dried by using oven, second group was by using vacuum freeze dryer methods. Dried carrot prepared from fresh carrot was analysed nutrient matter (energy, crude protein, crude oil, crude ash, beta carotene, mineral concentration and colour). The differences between groups in terms of energy, crude protein, ash, Ca and Mg was not significant (P>0,05). Crude oil, P, beta carotene content and colour values (L, a, b) with vacuum-freeze dryer group was greater than oven group (P<0,05). Consequently, carrot powder obtained by drying the vacuum-freeze dryer method can be used as a source of carotene. 

Keywords: Carrot, vacuum freeze dryer, oven, beta carotene.

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Authors: John R. Rose, James P. Cleveland, Alvin Fox

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Tandem mass spectrometry (MS/MS) is the engine driving high-throughput protein identification. Protein mixtures possibly representing thousands of proteins from multiple species are treated with proteolytic enzymes, cutting the proteins into smaller peptides that are then analyzed generating MS/MS spectra. The task of determining the identity of the peptide from its spectrum is currently the weak point in the process. Current approaches to de novo sequencing are able to compute candidate peptides efficiently. The problem lies in the limitations of current scoring functions. In this paper we introduce the concept of proteome signature. By examining proteins and compiling proteome signatures (amino acid usage) it is possible to characterize likely combinations of amino acids and better distinguish between candidate peptides. Our results strongly support the hypothesis that a scoring function that considers amino acid usage patterns is better able to distinguish between candidate peptides. This in turn leads to higher accuracy in peptide prediction.

Keywords: Tandem mass spectrometry, proteomics, scoring, peptide, de novo, mutual information

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Authors: Raza Hussain, Claire Gaiani, Joël Scher

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Dispersions of casein micelles (CM) were studied at a constant protein concentration of 5 wt % in high NaCl environment ranging from 0% to 12% by Dynamic light scattering (DLS) and Fourier Transform Infrared (FTIR). The rehydration profiles obtained were interpreted in term of wetting, swelling and dispersion stages by using a turbidity method. Two behaviours were observed depending on the salt concentration. The first behaviour (low salt concentration) presents a typical rehydration profile with a significant change between 3 and 6% NaCl indicating quick wetting, swelling and long dispersion stage. On the opposite, the dispersion stage of the second behaviour (high salt concentration) was significantly shortened indicating a strong modification of the protein backbone. A salt increase result to a destabilization of the micelle and the formation of mini-micelles more or less aggregated indicating an average micelles size ranging from 100 to 200 nm. For the first time, the estimations of secondary structural elements (irregular, ß-sheet, α-helix and turn) by the Amide III assignments were correlated with results from Amide I.

Keywords: Casein, DLS, FTIR, Ionic environment.

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Authors: Manuel A. Alonso-Tarajano, Roberto Mosca, Patrick Aloy

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Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase–pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed-specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Keywords: Kinase, phosphorylation, substrate specificity, adaptors, scaffolds, cellular colocalization.

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Authors: V. Barbakadze, L. Gogilashvili, L. Amiranashvili, M. Merlani, K. Mulkijanyan

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The high-molecular water-soluble preparations from several species of two genera (Symphytum and Anchusa) of Boraginaceae family Symphytum asperum, S. caucasicum, S.officinale and Anchusa italica were isolated. According to IR, 13C and 1H NMR, APT, 1D NOE, 2D heteronuclear 1H/13C HSQC and 2D DOSY experiments, the main chemical constituent of these preparations was found to be caffeic acid-derived polyether, namely poly[3-(3,4-dihydroxyphenyl)glyceric acid] (PDPGA) or poly[oxy-1- carboxy-2-(3,4-dihydroxyphenyl)ethylene]. Most carboxylic groups of this caffeic acid-derived polymer of A. italica are methylated.

Keywords: Anchusa, poly[3-(3, 4-dihydroxyphenyl)glyceric acid], poly[oxy-1-carboxy-2-(3, 4-dihydroxyphenyl)ethylene], Symphytum.

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Authors: Maria E. Manioudaki, Panayiota Poirazi

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Yeast cells live in a constantly changing environment that requires the continuous adaptation of their genomic program in order to sustain their homeostasis, survive and proliferate. Due to the advancement of high throughput technologies, there is currently a large amount of data such as gene expression, gene deletion and protein-protein interactions for S. Cerevisiae under various environmental conditions. Mining these datasets requires efficient computational methods capable of integrating different types of data, identifying inter-relations between different components and inferring functional groups or 'modules' that shape intracellular processes. This study uses computational methods to delineate some of the mechanisms used by yeast cells to respond to environmental changes. The GRAM algorithm is first used to integrate gene expression data and ChIP-chip data in order to find modules of coexpressed and co-regulated genes as well as the transcription factors (TFs) that regulate these modules. Since transcription factors are themselves transcriptionally regulated, a three-layer regulatory cascade consisting of the TF-regulators, the TFs and the regulated modules is subsequently considered. This three-layer cascade is then modeled quantitatively using artificial neural networks (ANNs) where the input layer corresponds to the expression of the up-stream transcription factors (TF-regulators) and the output layer corresponds to the expression of genes within each module. This work shows that (a) the expression of at least 33 genes over time and for different stress conditions is well predicted by the expression of the top layer transcription factors, including cases in which the effect of up-stream regulators is shifted in time and (b) identifies at least 6 novel regulatory interactions that were not previously associated with stress-induced changes in gene expression. These findings suggest that the combination of gene expression and protein-DNA interaction data with artificial neural networks can successfully model biological pathways and capture quantitative dependencies between distant regulators and downstream genes.

Keywords: gene modules, artificial neural networks, yeast, stress

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Authors: Majid Toghyani, Abbas Ali Gheisari, Ali Khodami, Mehdi Toghyani, Mohammad Mohammadrezaei, Ramin Bahadoran

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This experiment was conducted to investigate the effect of different levels of dietary chromium yeast (Cr-yeast) on thigh meat quality of broiler chicks reared under heat stress condition. Two hundred and forty Ross male chickens in heat stress condition (33±3°C) were allocated to five treatments in a completely randomized design. Treatments were supplemented with 0 (control), 200, 400, 800 and 1200 μg kg-1 Cr in the form of Cr yeast. Twelve chicks from each treatment were slaughtered at 42 d, to evaluate moisture, protein, lipid, pH and lipid oxidation of thigh meat. Protein, moisture, lipid and pH of thigh meat were not affected by supplemental Cr. Thigh meat lipid tended to decrease in broilers received 1200 μg kg-1. Storage time increased lipid oxidation of meat (P<0.01). Lipid oxidation of thigh muscle for two days of storage were affected by supplemental Cr and decreased (P<0.05). Results of this study showed that dietary Cr-yeast supplementation improved the thigh meat quality of broiler chicks in heat stress condition.

Keywords: Broiler, Heat stress, Chromium yeast, Thigh meat quality.

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Authors: Jae-Hong Kwon, Myung-Ho Chung, Tae-Yeon Oh, Hyeon-Seok Bae, Byeong-Kwon Ju

Abstract:

End-substitution of quarterthiophene and sexithiophene with hexyl groups leads to highly soluble conjugated oligomers,DZ-dihexylquarterthiophene (DH-4T) and DZ-dihexylsexithiophene (DH-6T). We have characterized these oligomers for optical and electrical properties. We fabricated an organic thin film transistor (OTFT) using the above two air-stable p-type organic semiconductor materials. We obtained a stable characteristic curve. The field effect mobility, Pwas calculated to be 3.2910-4 cm2/Vs for DH-6T based OTFT; while the DH-4T based OTFT had 1.8810-5 cm2/Vs.KeywordsOrganic thin film transistor, DZ-dihexylquarterthiophene, DZ-dihexylsexithiophene.

Keywords: Organic thin film transistor, DZ-dihexylquarterthiophene, DZ-dihexylsexithiophene.

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Authors: Permphan Dharmasaroja

Abstract:

Mutations of the telomeric copy of the survival motor neuron 1 (SMN1) gene cause spinal muscular atrophy. A deletion of the Eef1a2 gene leads to lower motor neuron degeneration in wasted mice. Indirect evidences have been shown that the eEF1A protein family may interact with SMN, and our previous study showed that abnormalities of neuromuscular junctions in wasted mice were similar to those of Smn mutant mice. To determine potential colocalization between SMN and tissue-specific translation elongation factor 1A2 (eEF1A2), an immunochemical analysis of HeLa cells transfected with the plasmid pcDNA3.1(+)C-hEEF1A2- myc and a new quantitative test of colocalization by intensity correlation analysis (ICA) was used to explore the association of SMN and eEF1A2. Here the results showed that eEF1A2 redistributed from the cytoplasm to the nucleus in response to serum and epidermal growth factor. In the cytoplasm, compelling evidence showed that staining for myc-tagged eEF1A2 varied in synchrony with that for SMN, consistent with the formation of a SMN-eEF1A2 complex in the cytoplasm of HeLa cells. These findings suggest that eEF1A2 may colocalize with SMN in the cytoplasm and may be a component of the SMN complex. However, the limitation of the ICA method is an inability to resolve colocalization in components of small organelles such as the nucleus.

Keywords: Intensity correlation analysis, intensity correlation quotient.

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Authors: Mardiana Ahamad Zabidi, Akmalluddin Md. Yunus

Abstract:

Soursop (Anona muricata) is one of the underutilized tropical fruits containing nutrients, particularly dietary fibre and antioxidant properties that are beneficial to human health. This objective of this study is to investigate the feasibility of matured soursop pulp flour (SPF) to be substituted with high-protein wheat flour in bread. Bread formulation was substituted with different levels of SPF (0%, 5%, 10% and 15%). The effect on physicochemical properties and sensory attributes were evaluated. Higher substitution level of SPF resulted in significantly higher (p<0.05) fibre, protein and ash content, while fat and carbohydrate content reduced significantly (p<0.05). FESEM showed that the bread crumb surface of control and 5% SPF appeared to distribute evenly and coalesced by thin gluten film. However, higher SPF substitution level in bread formulation exhibited a deleterious effect by formation of discontinuous gluten network. For texture profile analysis, 5% SPF bread resulted in the lowest value of hardness. The score of sensory evaluation showed that 5% SPF bread received good acceptability and is comparable with control bread.

Keywords: Bread, Physicochemical properties, Scanning electron microscopy (SEM), Sensory attributes, Soursop pulp flour.

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Authors: L. K. Davis

Abstract:

The effects of the evolution force are observable in nature at all structural levels ranging from small molecular systems to conversely enormous biospheric systems. However, the evolution force and work associated with formation of biological structures has yet to be described mathematically or theoretically. In addressing the conundrum, we consider evolution from a unique perspective and in doing so we introduce the “Fundamental Theory of the Evolution Force: FTEF”. We utilized synthetic evolution artificial intelligence (SYN-AI) to identify genomic building blocks and to engineer 14-3-3 ζ docking proteins by transforming gene sequences into time-based DNA codes derived from protein hierarchical structural levels. The aforementioned served as templates for random DNA hybridizations and genetic assembly. The application of hierarchical DNA codes allowed us to fast forward evolution, while dampening the effect of point mutations. Natural selection was performed at each hierarchical structural level and mutations screened using Blosum 80 mutation frequency-based algorithms. Notably, SYN-AI engineered a set of three architecturally conserved docking proteins that retained motion and vibrational dynamics of native Bos taurus 14-3-3 ζ.

Keywords: 14-3-3 docking genes, synthetic protein design, time based DNA codes, writing DNA code from scratch.

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Authors: Kati Manskinen, Risto Pöykiö, Hannu Nurmesniemi

Abstract:

In an assessment of the extractability of metals in green liquor dregs from the chemical recovery circuit of semichemical pulp mill, extractable concentrations of heavy metals in artificial gastric fluid were between 10 (Ni) and 717 (Zn) times higher than those in artificial sweat fluid. Only Al (6.7 mg/kg; d.w.), Ni (1.2 mg/kg; d.w.) and Zn (1.8 mg/kg; d.w.) showed extractability in the artificial sweat fluid, whereas Al (730 mg/kg; d.w.), Ba (770 mg/kg; d.w.) and Zn (1290 mg/kg; d.w.) showed clear extractability in the artificial gastric fluid. As certain heavy metals were clearly soluble in the artificial gastric fluid, the careful handling of this residue is recommended in order to prevent the penetration of green liquor dregs across the human gastrointestinal tract.

Keywords: Dregs, non-process elements, pulping, waste.

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

Abstract:

Various nanomaterials can be used as a drug delivery vehicles in nanomedicine, called nanocarriers. They can either be organic or inorganic, synthetic or natural-based. Although synthetic nanocarriers are easier to produce, they can often be toxic for the organism and thus not suitable for use in treatment. From naturalbased nanocarriers, the most commonly used are protein cages or viral capsids. In this work, virus bacteriophage λ was used for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2% maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000×g and 4°C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 μg·mL-1). Free drug molecules were removed by filtration. The encapsulation was verified using the absorbance for doxorubicin and atomic absorption spectrometry for platinum cytostatics. The amount of encapsulated drug linearly increased with the increasing concentration of applied drug with the determination coefficient R2=0.989 for doxorubicin; R2=0.967 for cisplatin; R2=0.989 for carboplatin and R2=0.996 for oxaliplatin. The overall encapsulation efficiency was calculated as 50% for doxorubicin; 8% for cisplatin; 6% for carboplatin and 10% for oxaliplatin.

Keywords: Bacteriophage λ, doxorubicin, platinum cytostatics, protein-based nanocarrier, viral capsid.

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