Search results for: protein folding

Authors: M. Michael Gromiha, Y. Yabuki, K. Imai, P. Horton, K. Fukui

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We have developed a database for membrane protein functions, which has more than 3000 experimental data on functionally important amino acid residues in membrane proteins along with sequence, structure and literature information. Further, we have proposed different methods for identifying membrane proteins based on their functions: (i) discrimination of membrane transport proteins from other globular and membrane proteins and classifying them into channels/pores, electrochemical and active transporters, and (ii) β-signal for the insertion of mitochondrial β-barrel outer membrane proteins and potential targets. Our method showed an accuracy of 82% in discriminating transport proteins and 68% to classify them into three different transporters. In addition, we have identified a motif for targeting β-signal and potential candidates for mitochondrial β-barrel membrane proteins. Our methods can be used as effective tools for genome-wide annotations.

Keywords: Membrane proteins, database, transporters, discrimination, β-signal.

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Authors: Tzong Yuan Wu, Sheng Kuo Hsieh, Tzyy Rong Jinn

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Fip-gts, an immunomodulatory protein purified from Ganoderma tsugae, has been reported to possess therapeutic effects in the treatment of cancer and autoimmune disease. For medicinal application, a recombinant Fip-gts was successfully expressed and purified in Sf21 insect cells by our previously work. It is important to evaluate the immunomodulatory activity of the rFip-gts. To assess the immunomodulatory potential of rFip-gts, the T lymphocytes of murine splenocytes were used in the present study. Results revealed that rFip-gts induced cellular aggregation formation. Additionally, the expression of IL-2 and IFN-r were up-regulated after the treatment of rFip-gts, and a corresponding increased production of IL-2 and IFN-r in a dose-dependent manner. The results showed that rFip-gts has an immunomodulatory activity in inducing Th1 lymphocytes from murine splenocytes released IL-2 and IFN-γ, thus suggest that rFip-gts may have therapeutic potential in vivo as an immune modulator.

Keywords: Fungal immunomodulatory protein, Ganodermatsugae, Interleukin 2, Interferon γ, Lingzhi.

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Authors: Almas Tayebi, Tayeb Saki Nejad, Alireza Shoukofar

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The trial in the city, located 170 kilometers from the Iranian city of Ahvaz was Omidiyeh. The main factor in this project includes 4 levels in control (without hormones), use of hormones in the seed, vegetative and flowering stage respectively. And sub-plots included 3 varieties of vetch in three levels, with local names, was the jewel in the study of light and Auxin in the vegetative and reproductive different times in different varieties of vetch was investigated. This test has been taken in the plots in a randomized complete block with four replications. In order to study the effects of the hormone Auxin in the growth stages (seed, vegetative and flowering) to control (no hormone Auxin) on three local varieties of vetch, the essence of light and plant height, number of pods per plant, seed number The pods, seeds per plant, grain weight, grain yield, plant dry weight and protein content were measured. Among the vetch varieties for plant height, number of pods per plant, a seed per plant, grain weight, grain yield, and plant dry weight and protein levels of 1 percent of plant and seed number per pod per plant at 5% level of There was no significant difference. Interactions for grain yield per plant, grain yield and protein levels of 1 percent and the number of seeds per pod and seed weight are significant differences in levels 5 and plant height and plant dry weight of the interaction were INFLUENCE There was no significant difference in them.

Keywords: Auxin hormones, various periods of growth, production components, vetch

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Authors: P. Grasgruber, J. Cacek, S. Hřebíčková

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The purpose of this study was to test the reliability of various standards that assess the quality of proteins via the “amino-acid score” and serve as a nutritional guideline for both children and adults. The height of young men in 42 European countries, Australia, New Zealand and USA was compared with the average consumption of food (after FAOSTAT, 2009) and a subsequent statistical analysis identified types of food with the most pronounced effect on physical growth. The results show that milk products and pork meat are by far the most significant nutritional factors in this regard. Cereals, vegetables and especially wheat played a strongly negative role. The results generally agreed best with the amino-acid score of proteins according to the standard of FAO 1985. In our opinion, the new standard of FAO 2007 underestimates the importance of tryptophan, which should provoke a debate about new modifications of the FAO guidelines.

Keywords: Protein quality, amino-acid score, physical growth, male height.

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Authors: T.A. Adegbola, IEA Aghachi, E.R. Sadiku

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To investigates the effect of fiberglass clamping process improvement on drape simulation prediction. This has great effect on the mould and the fiber during manufacturing process. This also, improves the fiber strain, the quality of the fiber orientation in the area of folding and wrinkles formation during the press-forming process. Drape simulation software tool was used to digitalize the process, noting the formation problems on the contour sensitive part. This was compared with the real life clamping processes using single and double frame set-ups to observe the effects. Also, restrains are introduced by using clips, and the G-clamps with predetermine revolution to; restrain the fabric deformation during the forming process.The incorporation of clamping and fabric restrain deformation improved on the prediction of the simulation tool. Therefore, for effective forming process, incorporation of clamping process into the drape simulation process will assist in the development of fiberglass application in manufacturing process.

Keywords: clamping, fiberglass, drape simulation, pressforming.

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Authors: A.C. Duarte, C.I.L. Justino, K. Duarte, A.C. Freitas, R. Pereira, P. Chaves, P. Bettencourt, S. Cardoso, T.A.P. Rocha-Santos

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This paper aims at overviewing the topics of a research project (CARDIOSENSOR) on the field of health sciences (biomaterials and biomedical engineering). The project has focused on the development of a nanosensor for the assessment of the risk of cardiovascular diseases by the monitoring of C-reactive protein (CRP), which has been currently considered as the best validated inflammatory biomarker associated to cardiovascular diseases. The project involves tasks such as: 1) the development of sensor devices based on field effect transistors (FET): assembly, optimization and validation; 2) application of sensors to the detection of CRP in standard solutions and comparison with enzyme-linked immunosorbent assay (ELISA); and 3) application of sensors to real samples such as blood and saliva and evaluation of their ability to predict the risk of cardiovascular disease.

Keywords: Carbon nanotubes field effect transistors, cardiovascular diseases, C-reactive protein, sensor.

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast.

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Authors: R. U. Hamzah, H. L. Muhammad, A. Sayyadi, M. B Busari, R. Garba, M. B. Umar, A. N Abubakar

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Lead (Pb) and Cadmium (Cd) are toxic, non-essential transition metals that pose many health risks for both humans and animals. They are environmental toxicants which contribute to testicular damage resulting to infertility problem among male populace worldwide. Chelating agents used for lead and cadmium toxicity are not readily available, toxic, expensive and unable to mop up most of the toxic metals accumulated in various organs. In this study, the effect of crude extract (CE), ethyl acetate fraction (EF) and acetone fraction (AF) of Pterocarpus mildbraedii leaf extract was assessed on cadmium-lead chloride induced testicular damaged in male albino Wistar rats. CE of the leaf was obtained by extracting in absolute methanol which was further subjected to solvent partitioning via vacuum liquid chromatographic (VLC) techniques using ethyl acetate, acetone and 70% methanol. A preliminary phytochemical screening and in vitro antioxidants guided activities on the CE and fractions were determined using standard methods. EF, AF and CE which exhibited significant in vitro activity were subjected to an in vivo study using Wistar rats. In vivo antioxidant markers, male reproductive hormones, testicular enzymes and DNA damage markers were analyzed on the rats’ testes supernatant. AF had the highest quantities of phenols (319.00 mg/g), flavonoids (8.87 mg/g) and tannins (8.87 mg/g) while methanol and EFs were richer in saponins (135.32 µg/g) and alkaloids (38.34 µg/g) respectively. A dose dependent 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric reducing antioxidant power (FRAP) and lipid peroxidation were observed in all the extract with high antioxidants power in CE and AF. Administration of lead-cadmium chloride solution significantly (p > 0.05) decreases the testicular superoxide dismutase (SOD) activity to 6.82 unit/mg protein, Catalase (CAT) activity to 8.07 of H2O2 consumed/unit/mg protein and Glutathione (GSH) concentration to 31.30 ug/mg protein. There was a concomitant increase in the level of Malondialdehyde (MDA) to a value of 23.70 mmol/mg protein. In addition, lead-cadmium chloride solution significantly (p > 0.05) increases the testicular marker enzymes (Alkaline phosphatase (119.57 u/L), lactate dehydrogenase (357.05 u/L), Acid phosphatase (98.65 u/L)) and DNA damage markers (conjugated dienes (93.39 nmol/mg protein), carbonyl protein (35.39 nmol/mg protein), DNA fragmentation percentage (32.12%)) with lowered testicular hormones (Testosterone (3.1 ng/mL), Follicle stimulating (0.35 IU/mL) and Luteinizing hormones (0.15 IU/mL)) of the animals in negative control group when compared with other treated groups. Treatment with Pterocarpus mildbraedii leaf extract reverts the observed changes with the best activities found in the CE and AFs in a dose dependent manner. Pterocarpus mildbraedii leaf extract ameliorated the lead/cadmium induced testicular damage in male albino rats. The restoration of the aforementioned parameters by some of the extract dosages were comparable to the standard drug with higher activities in the crude and AF. Therefore, Pterocarpus mildbraedii leaf extract can be explored further for the management of lead/cadmium induced toxicity.

Keywords: Cadmium, lead, Pterocarpus mildbraedii, testicular damage.

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Authors: N. Cetinkaya, N. H. Ozdemir

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The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinine levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in μmol/W0.75 (p<0.05). Mean creatinine values in μmol/W0.75 and allantoin:creatinine ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage:concentrate ratio in Kıvırcık Yearlings.

Keywords: Allantoin, creatinine, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisiae.

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Authors: Mohamed A. Ahmed, Lydia J. Campbell

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The fortified of soft wheat flour with cowpea flour in bread making was investigated. The Soft wheat flour (SWF) was substituted by cowpea flour at levels of 5, 15 and 20%. The protein content of composite breads ranged from 6.1 – 9.9%. Significant difference was observed in moisture, protein and crude fibre contents of control (wheat bread) and composite bread at 5% addition of cowpea. Water absorption capacities of composite flours increased with increasing levels of cowpea flour in the blend. The specific loaf volume decreased significantly with increased cowpea content of blends. The overall acceptability of the 5% cowpea flour content of composite bread was not significantly different from the control (Soft Wheat-bread) but there is significantly different with increasing the levels of cowpea flour in the blend more than 5%.

Keywords: Cowpea flour, wheat flour, baking properties, sensory quality.

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Authors: Rezvan Najafi, Korosh Heydari, Massoud Saidijam

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5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.

Keywords: Colorectal cancer, miR-200c, 5-FU resistance, E-cadherin, PTEN.

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Authors: Kasra Afsahi, Maryam Soheilifar

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Cerebrovascular accident (CVA) is a neurological side effect of COVID-19 disease wit high rate in pandemics. Effect of COVID-19 disease on disorder is unclear. In this cohort, patients with COVID-19 disease were assessed. 60 CVA cases were assessed in a referral hospital in 2020. The major factor was mortality and the cases were those with and without death. The groups were compared for all features about mortality in the patients with COVID-19 and CVA. Totally 23 out of 60 cases (38.3%) were expired. In univariate analysis there was significant association for death by ischemic heart disease (P = 0.015), high-severity stroke (P = 0.012), high C-reactive protein (CRP) (P = 0.001), high ESR (P = 0.009), pleural effusion (P = 0.005), pericardial effusion (P = 0.027), cardiomegaly (P = 0.005), ground glass opacity (P = 0.001), and consolidation (P = 0.001). Among these factors, there was significant association only for CRP (P = 0.001) and consolidation (P = 0.003) in multivariate analysis. Mortality in the cases with COVID-19-related CVA is one-third and it has relationship to elevated CRP and finding the consolidation in the computerized tomography scan of the lungs.

Keywords: COVID-19, stroke, prognosis, C-reactive protein, CRP.

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Authors: P. M. Ikonić, T. A. Tasić, L. S. Petrović, S. B. Škaljac, M. R. Jokanović, V. M. Tomović, B. V. Šojić, N. R. Džinić, A. M. Torbica, B. B. Ikonić

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The aim of the study was to determine how different ripening processes (traditional vs. industrial) influenced the proteolysis in traditional Serbian dry-fermented sausage Petrovská klobása. The obtained results indicated more intensive pH decline (0.7 units after 9 days) in industrially ripened products (I), what had a positive impact on drying process and proteolytic changes in these samples. Thus, moisture content in I sausages was lower at each sampling time, amounting 24.7% at the end of production period (90 days). Likewise, the process of proteolysis was more pronounced in I samples, resulting in higher contents of non-protein nitrogen (NPN) and free amino acids nitrogen (FAAN), as well as in faster and more intensive degradation of myosin (≈220 kDa), actin (≈45 kDa) and other polypeptides during processing. Consequently, the appearance and accumulation of several protein fragments were registered.

Keywords: Dry-fermented sausage, Petrovská klobása, proteolysis, ripening process.

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Authors: Mustafa Salman, Nurcan Cetinkaya, Zehra Selcuk, Bugra Genc

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The objectives of the present study were to estimate the microbial-N flow to the small intestine and to predict the digestible organic matter intake (DOMI) in grazing Karayaka sheep based on urinary excretion of purine derivatives (xanthine, hypoxanthine, uric acid, and allantoin) by the use of spot urine sampling under field conditions. In the trial, 10 Karayaka sheep from 2 to 3 years of age were used. The animals were grazed in a pasture for ten months and fed with concentrate and vetch plus oat hay for the other two months (January and February) indoors. Highly significant linear and cubic relationships (P<0.001) were found among months for purine derivatives index, purine derivatives excretion, purine derivatives absorption, microbial-N and DOMI. Through urine sampling and the determination of levels of excreted urinary PD and Purine Derivatives / Creatinine ratio (PDC index), microbial-N values were estimated and they indicated that the protein nutrition of the sheep was insufficient.

In conclusion, the prediction of protein nutrition of sheep under the field conditions may be possible with the use of spot urine sampling, urinary excreted PD and PDC index. The mean purine derivative levels in spot urine samples from sheep were highest in June, July and October. Protein nutrition of pastured sheep may be affected by weather changes, including rainfall. Spot urine sampling may useful in modeling the feed consumption of pasturing sheep. However, further studies are required under different field conditions with different breeds of sheep to develop spot urine sampling as a model.

Keywords: Karayaka sheep, spot sampling, urinary purine derivatives, PDC index, microbial-N, feed intake.

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Authors: Abdulqader M. Mohsen, Ahamad Tajudin Khader, Dhanesh Ramachandram, Abdullatif Ghallab

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The physical methods for RNA secondary structure prediction are time consuming and expensive, thus methods for computational prediction will be a proper alternative. Various algorithms have been used for RNA structure prediction including dynamic programming and metaheuristic algorithms. Musician's behaviorinspired harmony search is a recently developed metaheuristic algorithm which has been successful in a wide variety of complex optimization problems. This paper proposes a harmony search algorithm (HSRNAFold) to find RNA secondary structure with minimum free energy and similar to the native structure. HSRNAFold is compared with dynamic programming benchmark mfold and metaheuristic algorithms (RnaPredict, SetPSO and HelixPSO). The results showed that HSRNAFold is comparable to mfold and better than metaheuristics in finding the minimum free energies and the number of correct base pairs.

Keywords: Metaheuristic algorithms, dynamic programming algorithms, harmony search optimization, RNA folding, Minimum free energy.

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Authors: Le Thuy Mai, Vu Nguyen Thanh

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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: Jing Zhang, Lian Wu, Jie-ming Qu, Chun-xue Bai, Mervyn J Merrilees, Peter N Black

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COPD is characterized by loss of elastic fibers from small airways and alveolar walls, with the decrease in elastin increasing with disease severity. It is unclear why there is a lack of repair of elastic fibers. We have examined fibroblasts cultured from lung tissue from normal and COPD subjects to determine if the secretory profile explains lack of tissue repair. In this study, fibroblasts were cultured from lung parenchyma of bronchial carcinoma patients with varying degrees of COPD; controls (non-COPD, n=5), mild COPD (GOLD 1, n=5) and moderate-severe COPD (GOLD 2-3, n=12). Measurements were made of proliferation, senescence-associated beta-galactosidase-1, mRNA expression of IL-6, IL-8, MMP-1, tropoelastin and versican, and protein levels for IL-6, IL-8, PGE2, tropoelastin, insoluble elastin, and versican. It was found that GOLD 2-3 fibroblasts proliferated more slowly (p<0.01) and had higher levels of senescence-associated beta-galactosidase-1 (p<0.001) than controls (non-COPD). GOLD 2-3 fibroblasts showed significant increases in mRNA and/or protein for IL-6, IL-8, MMP-1, PGE2, versican (p<0.01) and tropoelastin (p<0.05). mRNA expression and/or protein levels of tropoelastin (p<0.01), versican (p<0.02), IL-6 (p<0.05) and IL-8 (p<0.05) were negatively correlated with FEV1%. Insoluble elastin was not increased. In summary, fibroblasts from moderate to severe COPD subjects display a secretory phenotype with up-regulation of inflammatory molecules including the matrix proteoglycan versican, and increased soluble, but not insoluble, elastin. Versican inhibits assembly of tropoelastin into insoluble elastin and we conclude that the pro-inflammatory phenotype of COPD fibroblasts it is not compatible with repair elastic fibers.

Keywords: COPD, pulmonary fibroblasts, pro-inflammatory phenotype, versican, elastin

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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

Abstract:

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

Abstract:

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: 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

Abstract:

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

Abstract:

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

Abstract:

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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