Search results for: whey protein concentrate

Authors: Aakansha Gupta, Neha Vadnere, Tapasvi Soni, M. Anbarsi

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Protein secondary structure prediction is one of the most important goals pursued by bioinformatics and theoretical chemistry; it is highly important in medicine and biotechnology. Some aspects of protein functions and genome analysis can be predicted by secondary structure prediction. This is used to help annotate sequences, classify proteins, identify domains, and recognize functional motifs. In this paper, we represent protein secondary structure as a mathematical model. To extract and predict the protein secondary structure from the primary structure, we require a set of parameters. Any constants appearing in the model are specified by these parameters, which also provide a mechanism for efficient and accurate use of data. To estimate these model parameters there are many algorithms out of which the most popular one is the EM algorithm or called the Expectation Maximization Algorithm. These model parameters are estimated with the use of protein datasets like RS126 by using the Bayesian Probabilistic method (data set being categorical). This paper can then be extended into comparing the efficiency of EM algorithm to the other algorithms for estimating the model parameters, which will in turn lead to an efficient component for the Protein Secondary Structure Prediction. Further this paper provides a scope to use these parameters for predicting secondary structure of proteins using machine learning techniques like neural networks and fuzzy logic. The ultimate objective will be to obtain greater accuracy better than the previously achieved.

Keywords: model parameters, expectation maximization algorithm, protein secondary structure prediction, bioinformatics

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Authors: Mohammed Hakim Jafferali, Balaje Vijayaraghavan, Ricardo A. Figueroa, Ellinor Crafoord, Veronica J. Larsson, Einar Hallberg, Santhosh Gudise

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The nuclear envelope which surrounds the chromatin of eukaryotic cells contains more than a hundred transmembrane proteins. Mutations in some genes encoding nuclear envelope proteins give rise to human diseases including neurological disorders. The function of many nuclear envelope proteins is not well established. This is partly because nuclear envelope proteins and their interactions are difficult to study due to the inherent resistance to extraction of nuclear envelope proteins. We have developed a novel method called MCLIP, to identify interacting partners of nuclear envelope proteins in live cells. Using MCLIP, we found three new binding partners of the inner nuclear membrane protein Samp1: the intermediate filament protein Lamin B1, the LINC complex protein Sun1 and the G-protein Ran. Furthermore, using in vitro studies, we show that Samp1 binds both Emerin and Ran directly. We have also studied the interaction between Samp1 and Ran in detail. The results show that the Samp1 binds stronger to RanGTP than RanGDP. Samp1 is the first transmembrane protein known to bind Ran and it is tempting to speculate that Samp1 may provide local binding sites for RanGTP at membranes.

Keywords: MCLIP, nuclear envelope, ran, Samp1

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Authors: Yi-Hui Lai, Chih-Hsiang Yang, Li-Chung Hsu

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The inflammasome is a protein complex that modulates caspase-1 activity, resulting in proteolytic cleavage of proinflammatory cytokines such as IL-1β and IL-18, into their bioactive forms. It has been shown that the inflammasomes play a crucial role in the clearance of pathogenic infection and tissue repair. However, dysregulated inflammasome activation contributes to a wide range of human diseases such as cancers and auto-inflammatory diseases. Yet, regulation of NLRP3 inflammasome activation remains largely unknown. We discovered a novel DEAD box protein, whose biological function has not been reported, not only negatively regulates NLRP3 inflammasome activation by interfering NLRP3 inflammasome assembly and cellular localization but also mitigate pyroptosis upon pathogen evasion. The DEAD-box protein is the first DEAD-box protein gets involved in modulation of the inflammasome activation. In our study, we found that caspase-1 activation and mature IL-1β production were largely enhanced upon LPS challenge in the DEAD box-containing protein- deleted THP-1 macrophages and bone marrow-derived macrophages (BMDMs). In addition, this DEAD box-containing protein migrates from the nucleus to the cytoplasm upon LPS stimulation, which is required for its inhibitory role in NLRP3 inflammasome activation. The DEAD box-containing protein specifically interacted with the LRR motif of NLRP3 via its DEAD domain. Furthermore, due to the crucial role of the NLRP3 LRR domain in the recruitment of NLRP3 to mitochondria and binding to its adaptor ASC, we found that the interaction of NLRP3 and ASC was downregulated in the presence of the DEAD box-containing protein. In addition to the mechanical study, we also found that this DEAD box protein protects host cells from inflammasome-triggered cell death in response to broad-ranging pathogens such as Candida albicans, Streptococcus pneumoniae, etc., involved in nosocomial infections and severe fever shock. Collectively, our results suggest that this novel DEAD box molecule might be a key therapeutic strategy for various infectious diseases.

Keywords: inflammasome, inflammation, innate immunity, pyroptosis

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Authors: Jamerson Felipe Pereira Lima, Jeane Cecília Bezerra de Melo

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The difficulty and cost related to obtaining of protein tertiary structure information through experimental methods, such as X-ray crystallography or NMR spectroscopy, helped raising the development of computational methods to do so. An approach used in these last is prediction of tridimensional structure based in the residue chain, however, this has been proved an NP-hard problem, due to the complexity of this process, explained by the Levinthal paradox. An alternative solution is the prediction of intermediary structures, such as the secondary structure of the protein. Artificial Intelligence methods, such as Bayesian statistics, artificial neural networks (ANN), support vector machines (SVM), among others, were used to predict protein secondary structure. Due to its good results, artificial neural networks have been used as a standard method to predict protein secondary structure. Recent published methods that use this technique, in general, achieved a Q3 accuracy between 75% and 83%, whereas the theoretical accuracy limit for protein prediction is 88%. Alternatively, to achieve better results, support vector machines prediction methods have been developed. The statistical evaluation of methods that use different AI techniques, such as ANNs and SVMs, for example, is not a trivial problem, since different training sets, validation techniques, as well as other variables can influence the behavior of a prediction method. In this study, we propose a prediction method based on artificial neural networks, which is then compared with a selected SVM method. The chosen SVM protein secondary structure prediction method is the one proposed by Huang in his work Extracting Physico chemical Features to Predict Protein Secondary Structure (2013). The developed ANN method has the same training and testing process that was used by Huang to validate his method, which comprises the use of the CB513 protein data set and three-fold cross-validation, so that the comparative analysis of the results can be made comparing directly the statistical results of each method.

Keywords: artificial neural networks, protein secondary structure, protein structure prediction, support vector machines

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Authors: Zunnu Raen Akhtar

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Cotton Leaf Curl Disease (CLCuD) is from Gemini virus and is transmitted through whiteflies in cotton. Transgenic cotton containing Antisense Coat Protein (ACP) has been found to show better results against CLCuD in cotton. In current research, Antisense Coat Protein was inserted in cotton plants to observe resistance developed in the cotton plants against CLCuD. T1 generation of plants were observed for its expression in plants. Tests were carried out to observe the expression of Antisense Coat Protein using Polymerase Chain Reaction (PCR) technique and by southern blotting. Whiteflies showing positive Cotton Leaf Curl Virus (CLCV) were reared and released in bioassay on ACP expressing cotton plants under laboratory as well as confined semi-field conditions. Results confirmed the expression of AC protein in PCR and southern blotting. Further laboratory results showed that cotton plants expressing AC protein showed rare incidence of CLCuD infection as compared to control. In the confined semi-field, similar results were observed in AC protein expressing cotton as compared to control. These results explicitly show that ACP can help to tackle the CLCuD issue in the future and further studies on biochemical processes involved in these plants and effects of ACP induction on non-target organisms should also be studied for eco-system.

Keywords: cotton, white flies, antisense coat protein, CLCV

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Authors: J. Suwanprateeb, F. Thammarakcharoen

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Calcium phosphate coating (CaP) has been employed for protein delivery, but the typical direct protein adsorption on the coating led to low incorporation content and fast release of the protein from the coating. By using bovine serum albumin (BSA) as a model protein, rapid biomimetic co-precipitation between calcium phosphate and BSA was employed to control the distribution of BSA within calcium phosphate coating during biomimetic formation on titanium surface for only 6 h at 50 oC in an accelerated calcium phosphate solution. As a result, the amount of BSA incorporation and release duration could be increased by using a rapid biomimetic co-precipitation technique. Up to 43 fold increases in the BSA incorporation content and the increase from 6 h to more than 360 h in release duration compared to typical direct adsorption technique were observed depending on the initial BSA concentration used during co-precipitation (1, 10, and 100 microgram/ml). From X-ray diffraction and Fourier transform infrared spectroscopy studies, the coating composition was not altered with the incorporation of BSA by this rapid biomimetic co-precipitation and mainly comprised octacalcium phosphate and hydroxyapatite. However, the microstructure of calcium phosphate crystals changed from straight, plate-like units to curved, plate-like units with increasing BSA content.

Keywords: biomimetic, Calcium Phosphate Coating, protein, titanium

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Authors: Alireza Naseri, Susan L. Holdt, Charlotte Jacobsen

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In 2014, the global amount of carrageenan production was 60,000 ton with a value of US$ 626 million. From this number, it can be estimated that the total dried seaweed consumption for this production was at least 300,000 ton/year. The protein content of these types of seaweed is 5 – 25%. If just half of this total amount of protein could be extracted, 18,000 ton/year of a high-value protein product would be obtained. The overall aim of this study was to develop a technology that will ensure further utilization of the seaweed that is used only as raw materials for carrageenan production as single extraction at present. More specifically, proteins should be extracted from the seaweed either before or after extraction of carrageenan with focus on maintaining the quality of carrageenan as a main product. Different mechanical, chemical and enzymatic technologies were evaluated. The optimized process was implemented in lab scale and based on its results; the new experiments were done a pilot and larger scale. In order to calculate the efficiency of the new upstream multi-extraction process, protein content was tested before and after extraction. After this step, the extraction of carrageenan was done and carrageenan content and the effect of extraction on yield were evaluated. The functionality and quality of carrageenan were measured based on rheological parameters. The results showed that by using the new multi-extraction process (submitted patent); it is possible to extract almost 50% of total protein without any negative impact on the carrageenan quality. Moreover, compared to the routine carrageenan extraction process, the new multi-extraction process could increase the yield of carrageenan and the rheological properties such as gel strength in the final carrageenan had a promising improvement. The extracted protein has initially been screened as a plant protein source in typical food applications. Further work will be carried out in order to improve properties such as color, solubility, and taste.

Keywords: carrageenan, extraction, protein, seaweed

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Authors: A. Lubis, R. Widayanti, Z. Hikmah, A. Endaryanto, A. Harsono, A. Harianto, R. Etika, D. K. Handayani, M. Sampurna

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Neonatal lupus erythematous (NLE) is a rare disease marked by clinical characteristic and specific maternal autoantibody. Many cutaneous, cardiac, liver, and hematological manifestations could happen with affect of one organ or multiple. In this case, both babies were premature, low birth weight (LBW), small for gestational age (SGA) and born through caesarean section from a systemic lupus erythematous (SLE) mother. In the first case, we found a baby girl with dyspnea and grunting. Chest X ray showed respiratory distress syndrome (RDS) great I and echocardiography showed small atrial septal defect (ASD) and ventricular septal defect (VSD). She also developed anemia, thrombocytopenia, elevated C-reactive protein, hypoalbuminemia, increasing coagulation factors, hyperbilirubinemia, and positive blood culture of Klebsiella pneumonia. Anti-Ro/SSA and Anti-nRNP/sm were positive. Intravenous fluid, antibiotic, transfusion of blood, thrombocyte concentrate, and fresh frozen plasma were given. The second baby, male presented with necrotic tissue on the left ear and skin rashes, erythematous macula, athropic scarring, hyperpigmentation on all of his body with various size and facial haemorrhage. He also suffered from thrombocytopenia, mild elevated transaminase enzyme, hyperbilirubinemia, anti-Ro/SSA was positive. Intravenous fluid, methyprednisolone, intravenous immunoglobulin (IVIG), blood, and thrombocyte concentrate transfution were given. Two cases of neonatal lupus erythematous had been presented. Diagnosis based on clinical presentation and maternal auto antibody on neonate. Organ involvement in NLE can occur as single or multiple manifestations.

Keywords: neonatus lupus erythematous, maternal autoantibody, clinical characteristic, multiple organ manifestation

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Authors: Kusmartono, Siti Chuzaemi, Hartutik dan Mashudi

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Two studies were conducted involving an in vitro (Expt 1) and in vivo (Expt 2) measurements. Expt 1. aimed to evaluate effects of adding CT extracts on gas production and efficiency of microbial protein synthesis (EMPS), Expt 2 aimed to evaluate effects of supplementing shrub/tree leaves as CT source on feed consumption, digestibility, N retention, body weight gain and dressing percentage of growing sheep fed on elephant grass (EG) as a basal diet.Ten shrub and tree leaves used as CT sources were wild sunflower (Tithonia diversifolia), mulberry (Morus macroura), cassava (Manihot utilissima), avicienna (Avicennia marina), calliandra (Calliandra calothyrsus), sesbania (Sesbania grandiflora), acacia (acacia vilosa), glyricidia (Glyricidia sepium), jackfruit (Artocarpus heterophyllus), moringa (Moringa oleifera). The treatments applied in Expt 1 were: T1=Elephant grass (60%)+concentrate (40%); T2 = T1 + CT (3% DM); T3= T2 + PEG; T4 = T1 + CT (3.5% DM); T5 = T4 + PEG; T6 = T1 + CT (4% DM) and T7 = T6 + PEG. Data obtained were analysed using Randomized Block Design. Statistical analyses showed that treatments significanty affected (P<0.05) total gas production and EMPS. The lowest values of total gas production (45.9 ml/500 mg DM) and highest value of EMPS (64.6 g/kg BOTR) were observed in the treatment T4 (3.5% CT from cassava leave extract). Based on this result it was concluded that this treatment was the best and was chosen for further investigation using in vivo method. The treatmets applied for in vivo trial were: T1 = EG (60%) + concentrate (40%); T2 = T1 + dried cassava leave (equivalent to 3.5% CT); T3 = T2 + PEG. 18 growing sheep aging of 8-9 months and weighing of 23.67kg ± 1.23 were used in Expt 2. Results of in vivo study showed that treatments significanty affected (P<0.05) nutrients intake and digestibility (DM, OM and CP). N retention for sheep receiving treatment T2 were significantly higher (P<0.05; 15.6 g/d) than T1 (9.1 g/d) and T3 (8.53 g/d). Similar results were obtained for daily weight gain where T2 were the highest (62.79 g/d), followed by T1 (51.9 g/d) and T3 (52.85 g/d). Dressing percentage of T2 was the highest (51.54%) followed by T1 (49.61%) and T3 (49.32%). It can be concluded that adding adding dried cassava leaves did not reduce palatability due to CT, but rather increased OM digestibility and hence feed consumption was improved. N retention was increased due to the action of CT in the cassava leaves and this may have explained a higher input of N into duodenum which was further led to higer daily weight gain and dressing percentage.

Keywords: in vitro gas production, sheep, shrub and tree leaves, condensed tannin

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Authors: Ji-Hyon Kil, Kew-Cheol Shim, Kyoung-Ae Park, Kyoungho Kim

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Ambrosia trifida L. is designated as invasive alien species by the Act on the Conservation and Use of Biodiversity by the Ministry of Environment, Korea. The purpose of present paper was to investigate the inhibitory effects of aqueous extracts of A.trifida on the development of root hairs of Triticum aestivum L., and Allium tuberosum Rottler ex Spreng and the electrophoretic protein patterns of their radicles. The development of root hairs was inhibited by increasing of aqueous extract concentrations. Through SDS-PAGE, the electrophoretic protein bands of extracted proteins from their radicles were appeared in controls, but protein bands of specific molecular weight disappeared or weakened in treatments. In conclusion, inhibitory effects of A. trifida made two receptor species changed morphologically, and at the molecular level in early growth stage.

Keywords: Ambrosia trifida L., invasive alien species, inhibitory effect, root hair, electrophoretic protein, radicle

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Authors: Birgul Hizlar, Sibel Karakaya

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Nowadays, consumers are more and more aware of the benefits beyond basic nutrition provided by food and food compounds. Between these, carotenoids have been demonstrated to exhibit multiple health benefits (for example, some types of cancer, cardiovascular diseases, eye disorders, among others). However, carotenoid bioaccessibility and bioavailability is generally rather low due to their specific localization in plant tissue and lipophilic nature. This situation is worldwide issue, since both developed and developing countries have their interest and benefits in increasing the uptake of carotenoids from the human diet. Recently, a new class of foods designed to improve the bioaccessibility/bioavailability of orally administered bioactive compounds is introduced: excipient foods. Excipient foods are specially designed foods which are prepared depending on the physicochemical properties of target bioactive compounds and increasing the bioavailability or bioaccessibility of bioactive compound. In this study, effects of food matrix (greating, boiling and mashing) and different excipient foods (olive oil, lemon juice, whey curd and dried artichoke leaf powder) on bioaccessibility of β-carotene in carrot were investigated by means of simulating in vitro gastrointestinal (GI) digestion. β-carotene contents of grated, boiled and mashed (after boiling process) carrots were 79.28, 147.63 and 151.19 μg/g respectively. No significant differences among boiled and mashed samples indicated that mashing process had no effect on the release of β-carotene from the food matrix (p > 0.05). On the contrary, mashing causes significant increase in the β-carotene bioaccessibility (p < 0.05). The highest β-carotene content was found in the mashed carrots incorporated with olive oil and lemon juice (C2). However, no significant differences between that sample and C1 (mashed carrot with lemon juice, olive oil, dried artichoke leaf powder), C3 (mashed carrot with addition of olive oil, lemon juice, whey curd) and). Similarly, the highest β-carotene bioaccessibility (50.26%) was found mashed C3 sample (p < 0.05). The increase in the bioaccessibility was approximately 5 fold and 50 fold when compared to grated and mashed samples containing olive oil, lemon juice and whey curd. The results demonstrate that both, food matrix and excipient foods, are able to increase the bioaccessibility of β-carotene.

Keywords: bioaccessibility, carotenoids, carrot, β-carotene

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Authors: Owonikoko A. D., Odoje O. F.

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Fresh sample of Eryngium alpinum was purchased and processed for leaf protein concentrates with a view to evaluating its nutritional potential, mineral composition, amino acid characteristics and phytochemical constituents. Using standard analytical methods. The proximate composition of the leaf protein concentrates revealed moisture content;(5.35±0.21)g/100g, ash;(11.37±0.43)g/100g, crude protein;(48.17±0.46)g/100g, crude fat;(15.38±0.07)g/100g, crude fibre (3.05±0.46)g/100g, and Nitrogen free extractive; (16.68±0.30) g/100g. The mineral content was: Na;(51.88±0.23) mg/100g, K;(65.40±0.32)mg/100g, Ca; (86.89±0.46)mg/100g, Mg;(49.27±0.42) mg/100g, Zn;(0.62±0.03)mg/100g, Fe (6.65±0.43)mg/100g, Mn;(0.96±0.54)mg/100g, Cd;(0.28±0.04)mg/100g, P; (8.55±0.97)mg/100g, while selenium, lead and mercury were not detected in the sample indicating that the sample is free of causing risk of metal poisoning. The results of phytochemical constituents showed phytate; (18.34±0.36)mg/100g, flavonoid (0.25±0.41)mg/100g. The sample contain both essential and non-essential amino acid, with the highest value of Glutamic acid (12.26) and the lowest value of Tryptophan 1.05. the content of the leaf protein content shows that the sample is fit for dietary consumption and could as well be processed to be used as food additives.

Keywords: mineral composition, phytochemical analysis, leaf protein concentrates, eryngium alpinum

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Authors: Illahi Bakhsh Marghazani, Nasrullah, Masood Ul Haq Kakar, Abdul Hameed Baloch, Ahmad Nawaz Khoso, Behram Chacher

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Protein sources are the major part of ration fed to dairy buffaloes in Pakistan however, the limited availability and lack of judicious use of protein resources are further aggravating the conditions to enhance milk and meat production. In order to gain maximum production from limited protein source availability, it is necessary to balance feed for rumen degradable and rumen undegradable protein fractions. This study planned to know the rumen degradable and rumen undegradable fractions in all vegetable protein sources with (formaldehyde and heat treatment) and without treatments. Samples of soybean meal, corn gluten meal 60%, maize gluten feed, guar meal, sunflower meal, rapeseed meal, rapeseed cake, canola meal, cottonseed cake, cottonseed meal, coconut cake, coconut meal, palm kernel cake, almond cake and sesame cake were collected from ten different geographical locations of Pakistan. These samples were also subjected to formaldehyde (1% /100g CP of test feed) and heat treatments (1 hr at 15 lb psi/100 g CP of test feed). In situ technique was used to know the ruminal degradability characteristics. Data obtained were fitted to Orskove equation. Results showed that both treatments significantly (P < 0.05) decreased ruminal degradability in all vegetable protein sources than untreated vegetable protein sources, however, of both treatments, heat treatment was more effective than formaldehyde treatment in decreasing ruminal degradability in most of the studied vegetable protein sources.

Keywords: formaldehyde and heat treatments, in situ technique, rumen degradable and rumen undegradable fractions, vegetable protein sources

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Authors: Abrha Reta, Ajebu Nurfeta, Genet Mengistu, Mohammed Beyan

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Fodder trees such as Ziziphus spina-christi have the potential to enhance the utilization of natural grazing resources and also to mitigate seasonal feed shortages. The experiment was conducted with the objective of evaluating the effect of supplementing Ziziphus spina-christi leaf meal (ZSCLM) to natural pasture hay on feed intake, body weight gain, digestibility, and carcass characteristics of Tigray highland sheep. A randomized complete block design was employed with 5 blocks based on initial body weight, and sheep were randomly assigned to five treatments. Treatments were: 100g concentrate mix + ad libtum natural pasture hay (T1), T1+ 100g ZSCLM (T2), T1 + 200g ZSCLM (T3), T1 + 300g ZSCLM (T4), and T1 + 400g ZSCLM (T5) on dry matter (DM) basis. Dry matter intake was greater (P<0.05) in sheep on T5 compared to T3 and T1, while the total DM intake among T2, T4, and T5 were similar. Crude protein and metabolizable energy intake differed (P<0.05) among treatments with highest and lowest values in T5 and T1, respectively. Average daily gain was higher (P<0.05) in sheep kept on T2, T3, and T4 diets than T1. Higher (P<0.05) DM digestibility was found in T4 and T5 than T1. The highest (P<0.05) OM and CP digestibility was observed in sheep fed T3, T4, and T5 diets. Rib eye muscle area was higher (P<0.05) for T4 than T1 and T2. Dressing percentage was similar (P>0.05) among treatments. The current study indicated that supplementation of Tigray highland sheep with 200g air-dried Ziziphus spina-christi leaf meal leaves with 100g of concentrate mixture in their diet significantly increased feed intake and apparent digestibility, body weight gain, hot carcass weight, and rib eye muscle area by improving feed conversion efficiency.

Keywords: body weight, carcass, digestibility, and ziziphus spina-christi leaf meal

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Authors: Amel Benhadji, Mourad Taleb Ahmed, Rachida Maachi

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

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

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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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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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

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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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Authors: B. Coşgun, O. Ozturk

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Canola is a specific edible type of rapeseed, developed in the 1970s, which contains about 40 percent oil. This research was carried out to determine the yield and some quality characteristics of some winter canola cultivars during the 2010-2011 vegetation period in Central Anatolia of Turkey. In this research; Oase, Dante, Californium, Excalibur, Elvis, ES Hydromel, Licord, Orkan, Vectra, Nelson, Champlain and NK Petrol winter canola varieties were used as material. The field experiment was set up in a “Randomized Complete Block Design” with three replications on 21 September 2010. In this research; seed yield, oil content, protein content, oil yield and protein yield were examined. As a result of this research; seed yield, oil content, oil yield and protein yield (except protein content) were significant differences between the cultivars. The highest seed yield (6348 kg ha-1) was obtained from the NK Petrol, while the lowest seed yield (3949 kg ha-1) was determined from the Champlain cultivar was obtained. The highest oil content (46.73%) was observed from Oase and the lowest value was obtained from Vectra (41.87%) cultivar. The highest oil yield (2950 kg ha-1) was determined from NK Petrol while the least value (1681 kg ha-1) was determined from Champlain cultivar. The highest protein yield (1539.3 kg ha-1) was obtained from NK Petrol and the lowest protein yield (976.5 kg ha-1) was obtained from Champlain cultivar. The main purpose of the cultivation of oil crops, to increase the yield of oil per unit area. According the result of this research, NK Petrol cultivar which ranks first with regard to both seed yield and oil yield between cultivars as the most suitable winter canola cultivar of local conditions.

Keywords: rapeseed, cultivar, seed yield, crude oil ratio, crude protein ratio, crude oil yield, crude protein yield

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Authors: Yafri Hazbi, Zaenal Bachruddin, Nafiatul Umami, Lies Mira Yusiati

Abstract:

The objective of this study was to identify the benefits of total mixture concentrate based on tofu waste silage (TMC-TWS) as ration containing lactic acid bacteria on performance of lambs. Fifteen weaning lambs (2-3 months old) were randomly divided into two treatment groups, treatment group I (TI) was fed with TMC-TWS as ration and treatment group II (TII) was fed with TMC-TWS fresh (without silage fermentation). The performance of lambs was evaluated on day 0, 15, and 30 to have data of body weight per day. Meanwhile, blood sampling and feces were made on the 30th day to get an analysis on the blood profile (erythrocytes (mill/ml), hemoglobin (g/dL), packed cell volume (%), and leukocytes (mill/ml)) and the number of worm eggs in feces. The results of this study showed no significant difference between the effect of different feed on the blood profile (erythrocytes (mill/ml), hemoglobin (g/dL), packed cell volume (%), as well as the number of worm eggs in the feces. However the results showed significant differences if it is low (P<0.05) due to the treatment group based on sex on body weight gain per day, feed conversion rate and the number of erythrocytes.

Keywords: lambs, total mixture concentrate, silage, acid lactid bacteria, blood profile, eggs worm in feces

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Authors: Ali Taheri Mirghaed, Sara Ahani, Ashkan Zargar, Seyyed Morteza Hoseini

Abstract:

Diseases are the major challenges in intensive aquaculture that cause significant annual losses. Antibiotic-therapy is a common way to control bacterial disease in fish, and oxytetracycline (OTC) is the only oral antibiotic in aquaculture approved FDA. OTC has been found to have negative effects on fish, such as oxidative stress and immune-suppression, thus, it is necessary to mitigate such effects. Medicinal herbs have various benefits on fish, including antioxidant, immunostimulant, and anti-microbial effects. Therefore, we hypothesized if dietary ginger meal (GM) interacts with dietary OTC by monitoring plasma protein fractions in rainbow trout. The study was conducted as a 2 × 2 factorial design, including diets containing 0 and 1% GM and 0 and 1.66 % OTC (corresponding to 100 mg/kg fish biomass per day). After ten days treating the fish (60 g individual weight) with these feeds, blood samples were taken from al treatments (n =3). Plasma was separated by centrifugation, and protein fractions were determined by electrophoresis. The results showed that OTC and GM had interaction effects on total protein (P<0.001), albumin (P<0.001), alpha-1 fraction (P=0.010), alpha-2 fraction (P=0.001), beta-2 fraction (P=0.014), and gamma fraction (P<0.001). Beta-1 fraction was significantly (P=0.030) affected by dietary GM. GM decreased plasma total protein, albumin, and beta-2 but increased beta-1 fraction. OTC significantly decreased total protein (P<0.001), albumin (P=0.001), alpha-2 fraction (P<0.001), beta-2 fraction (P=0.004), and gamma fraction (P<0.001) but had no significant effects on alpha-1 and beta-1 fractions. Dietary GM inhibited/suppressed the effects of dietary OTC on the plasma total protein and protein fractions. In conclusion, adding 1% GM to diet can mitigate the negative effects of dietary OTC on plasma proteins. Thus, GM may boost health of rainbow trout during the period of medication with OTC.

Keywords: ginger, plasma protein electrophoresis, dietary additive, rainbow trout

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Authors: Novriyanti Lubis, Riska Prasetiawati, Nuriani Rahayu

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The effectivity of lime juice on the cooked rice’s shelf-life was investigated. This research was proposed to get the optimal condition, such as concentration lime juice as the preservatives, and shelf-life cooked rice’s container to store using rice warmer. The effectivity was analysed total colony bacteriology, and physically. The variation of lime juice’s concentration that have been used were 0%, 0,46%, 0,93%, 1,40%, and 1,87%. The observation of cooked rice’s quality was done every 12 hours, including colour, smell, flavour, and total colony every 24 hours. Based on the result of the research considered from the cooked rice’s quality through observing the total of the colony bacteriology and physically, it showed the optimum concentrate which is effective preserve the cooked rise’s level concentrate was 0.93%.

Keywords: bacteriology, cooked rice's, lime juice, preservative

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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

Procedia PDF Downloads 287

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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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 139

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

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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

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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

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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 385