Search results for: neem leaf protein

Authors: Anna Trusek-Holownia, Andrzej Noworyta

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Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.

Keywords: peptide bond hydrolysis, kinetics, enzyme specificity, biologically active peptides

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Authors: Saeed S. Al-Sokari, Nasser A. Awadh Ali, Lianet Monzote

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Leishmaniasis (CL) is endemic in at least 82 countries and considered to be a major public-health problem (1). The annual incidence of CL is 1–1.5 million cases of which 90% occur in only seven countries: Afghanistan, Algeria, Brazil, Iran, Peru, Saudi Arabia and Syria (2). In Saudi Arabia, the disease was first described in 1973 by Moursy and Shoura (3). Currently, CL is common in the human population in different localities, including the Eastern Province of Saudi Arabia and in particular the Al-Hassa Oasis that is a known endemic area for CL (4). Five methanolic extracts obtained from Achillea biebersteinii (flower leaf), Euphorbia antiquorm, Solanum incanum (leaf and fruit extracts), collected from Albaha region and selected from ethno-botanical data, were screened for their anti-leishmanial activity against Leishmania amazonensis (6). The cytotoxic activity against normal peritoneal macrophages from normal BALB/c mice was also determined (6). The five extracts had IC50 values ranging from < 12.5 to 37.8 µg/ml against promastigotes. Achillea biebersteinii flower, Euphorbia antiquorm, Solanum incanum leaf extracts showed anti-leishmanial activities with IC50 between < 12.5 - 26.9µg/mL and acceptable selectivity indices of 8 - 5.

Keywords: plant extracts, Albaha, Leishmania amazonensis, Medicinal

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Authors: Huzaifa Bilal

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Russian wheat aphid (Diuraphis noxia, Kurdjumov) is considered an economically important wheat (Triticum aestivum L.) pest worldwide and in South Africa. The RWA damages wheat plants and reduces annual yields by more than 10%. Even though pest management by pesticides and resistance breeding is an attractive option, chemicals can cause harm to the environment. Furthermore, the evolution of resistance-breaking aphid biotypes has out-paced the release of resistant cultivars. An alternative strategy to reduce the impact of aphid damage on plants, such as priming, which sensitizes plants to respond effectively to subsequent attacks, is necessary. In this study, wheat plants at the seedling and flag leaf stages were primed by salicylic acid and isolate representative of two races of the leaf rust pathogen Puccinia triticina Eriks. (Pt), before RWA (South African RWA biotypes 1 and 4) infestation. Randomized complete block design experiments were conducted in the greenhouse to study plant-pest interaction in primed and non-primed plants. Analysis of induced aphid damage indicated salicylic acid differentially primed wheat cultivars for increased resistance to the RWASA biotypes. At the seedling stage, all cultivars were primed for enhanced resistance to RWASA1, while at the flag leaf stage, only PAN 3111, SST 356 and Makalote were primed for increased resistance. The Puccinia triticina efficaciously primed wheat cultivars for excellent resistance to RWASA1 at the seedling and flag leaf stages. However, Pt failed to enhance the four Lesotho cultivars' resistance to RWASA4 at the seedling stage and PAN 3118 at the flag leaf stage. The induced responses at the seedling and flag leaf stages were positively correlated in all the treatments. Primed plants induced high activity of antioxidant enzymes like peroxidase, ascorbate peroxidase and superoxide dismutase. High antioxidant activity indicates activation of resistant responses in primed plants (primed by salicylic acid and Puccina triticina). Isolates of avirulent Pt races can be a worthy priming agent for improved resistance to RWA infestation. Further confirmation of the priming effects needs to be evaluated at the field trials to investigate its application efficiency.

Keywords: Russian wheat aphis, salicylic acid, puccina triticina, priming

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Authors: Fellah Sihem

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The optimal use of soil moisture by culture, is related to the leaf area index, which stood in the cycle and its modulation according to the prevailing stress intensity. For a given stock of water in the soil, cultivar adapted and saving water is one that is no luxury consumption during the preanthesis. It modulates the leaf area index to regulate sweating in the degree of its water supply. In plants water saving, avoidance of dehydration is related to the reduction of water loss by cuticular and stomatal pathways. Muchow and Sinclair reported that the test of relative water content (TRE) is considered the best indicator of leaf water status. The search for indicators of the ability of the plant to make good use of the water, under water stress is a prerequisite for progress in improving performance under water stress. This experiment aims to characterize a set of durum wheat varieties, tested jars and vegetation under different levels of water stress to the surface of the leaf, relative water content, cell integrity, the accumulated biomass and efficiency of water use. The experiment was conducted during the 2005/2006 academic year, at the Agricultural Research Station of the Field Crop Institute of Setif, under semi-controlled conditions. Five genotypes of durum wheat (Triticum durum Desf) were evaluated for their ability to tolerate moderate and severe water stress. The results showed that geno types respond differently to water stress. Dry matter accumulation and growth rate varied among geno types and were significantly reduced. At severe water stress biomass accumulated by Boussalam was the least affected.

Keywords: water stress, triticum durum, biomass, cell membrane integrity, relative water content

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Authors: Faezeh Zaefarian1, Sajedeh Golmohammadzadeh, Mohammad Rezvani

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Weed management decisions for weed species can be derived from knowledge of seed germination biology. Experiments were conducted in laboratory and greenhouse to determine the effects of light, temperature, salt and water stress, seed burial depth on seed germination and seedling emergence of Papaver rhoeas and P.dubium and to assay the response of these species to commonly available POST herbicides. Germination of the Papaver seeds was influenced by the tested temperatures (day/night temperatures of 20 and 25 °C) and light. The concentrations of sodium chloride, ranging from 0 to 80 mM, influence germination of seeds. The osmotic potential required for 50% inhibition of maximum germination of P. rhoeas was -0.27 MPa and for P. dubium species was 0.25 MPa. Seedling emergence was greatest for the seeds placed at 1 cm and emergence declined with increased burial depth in the soil. No seedlings emerged from a burial depth of 6 cm. The herbicide 2,4-D at 400 g ai ha-1 provided excellent control of both species when applied at the four-leaf and six-leaf stages. However, at the six-leaf stage, percent control was reduced. The information gained from this study could contribute to developing components of integrated weed management strategies for Papaver species.

Keywords: germination, papaver species, planting depth, POST herbicides

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Authors: K. Uteulin, S. Mukhambetzhanov, I. Rakhimbaiev

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This experiment was performed to optimize the medium for tissue culture of Taraxacum kok-saghyz Rodin. Different tissue culture approaches such as shoot regeneration from seed, callus formation from leaf explants and plant regeneration from callus were investigated in this study. All the explants were cultured on MS basal medium supplemented with 20 g/l sucrose, 7 g/l agar and different plant growth regulators. Seeds of Taraxacum kok-saghyz were cultured on media containing different levels of BA and 2,4-D (0,5 and 1,0 and 3,0 mg/L) to direct shoot regeneration study. Leaf explants were cultured in different combination of BA (at three levels: 0.5, 1.0 and 3.0 mg/L) and zeatin (at two levels: 0.5 and 1.0 mg/L) to examine callus formation. After the callus formation the formed calli were cultured on different combinations of BA and NAA for shoot regeneration. BA at three levels (0.5 and 1.0 and 3.0 mg/L) and NAA at two levels (0.5 and 1.0 mg/L) in all possible combinations were used for shoot regeneration from callus. The results showed that the treatment containing 1.0 mg/L 2,4-D in combination with 1.0 mg/L BA was found to be the best one for shoot regeneration from seeds. The treatment with 1.0 mg/L BA in combination with 1.0 mg/L zeatin were found to be suitable treatments for callus production from leaf explants, as well. Moreover, 0.5 mg/L BA alone or in combination with 1.0 mg/L NAA were found to be the best treatments for shoot regeneration from callus.

Keywords: Taraxacum kok-saghyz Rodin, shoot regeneration, callus, plant

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Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske

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Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.

Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques

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Authors: M. Irfan Fareed, Muhammad Tahir, Alvina Gul Kazi

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Castor bean (Ricinus communis; family Euphorbiaceae) is cultivated for the production of oil and as an ornamental plant throughout tropical regions. Leaf samples from castor bean plants with leaf curl and vein thickening were collected from areas around Okara (Pakistan) in 2011. PCR amplification using diagnostic primers showed the presence of a begomovirus and subsequently the specific pair (BurNF 5’- CCATGGTTGTGGCAGTTGATTGACAGATAC-3’, BurNR 5’- CCATGGATTCACGCACAGGGGAACCC-3’) was used to amplify and clone the whole genome of the virus. The complete nucleotide sequence was determined to be 2,759 nt (accession No. HE985227). Alignments showed the highest levels of nucleotide sequence identity (98.8%) with Cotton leaf curl Burewala virus (CLCuBuV; accession No. JF416947) No. JF416947). The virus in castor beans lacks on intact C2 gene, as is typical of CLCuBuV in cotton. An amplification product of ca. 1.4 kb was obtained in PCR with primers for betasatellites and the complete nucleotide sequence of a clone was determined to be 1373 nt (HE985228). The sequence showed 96.3% nucleotide sequence identity to the recombinant Cotton leaf curl Multan betasatellite (CLCuMB; JF502389). This is the first report of CLCuBuV and its betasatellite infecting castor bean, showing this plant species as an alternate host of the virus. Already many alternate host have been reported from different alternate host like tobacco, tomato, hibiscus, okra, ageratum, Digera arvensis, habiscus, Papaya and now in Ricinus communis. So, it is suggested that these alternate hosts should be avoided to grow near cotton growing regions.

Keywords: Ricinus communis, begomovirus, betasatellite, agriculture

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Authors: B. F. Olanipekun, O. J. Oyelade, C. O. Osemobor

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Pigeon pea is a very good source of protein and micronutrient, but it is being underutilized in Nigeria because of several constraints. This research considered the effect of different processing methods on the quality attributes of pigeon pea used in bread production towards enhancing its utility. Pigeon pea was obtained at a local market and processed into the flour using three processing methods: soaking, sprouting and roasting and were used to bake bread in different proportions. Chemical composition and sensory attributes of the breads were thereafter determined. The highest values of protein and ash contents were obtained from 20 % substitution of sprouted pigeon pea in wheat flour and may be attributable to complex biochemical changes occurring during hydration, to invariably lead to protein constituent being broken down. Hydrolytic activities of the enzymes from the sprouted sample resulted in improvement in the constituent of total protein probably due to reduction in the carbohydrate content. Sensory qualities analyses showed that bread produced with soaked and roasted pigeon pea flours at 5 and 10% inclusion, respectively were mostly accepted than other blends, and products with sprouted pigeon pea flour were least accepted. The findings of this research suggest that supplementing wheat flour with sprouted pigeon peas have more nutritional potentials. However, with sensory analysis indices, the soaked and roasted pigeon peas up to 10% are majorly accepted, and also can improve the nutritional status. Overall, this will be very beneficial to population dependent on plant protein in order to combat malnutrition problems.

Keywords: pigeon pea, processing, protein, malnutrition

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Authors: Maria Magdy Danial Riad

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The objective of this study was to investigate the effect of different protein sources with different degradability ratios during late gestation of ewes on colostrum composition and its IgG concentration, body weight change of dams, and birth weight of their lambs. Objectives: 35 multiparous native crossbred ewes (BW= 59±2.5kg) were randomly allocated to five dietary treatments (7 ewes / treatment) for 2 months prior to lambing. Methods: Experimental diets were isonitrogenous (12.27% CP) and isocaloric (2.22 Mcal ME/kg DM). In diet I (the control), solvent extract soybeans (SESM 33% RUP of CP), II feed grade urea (FGU 31% RUP), III slow release urea (SRU 31% RUP). As sources of undegradable protein, extruded expeller SBM-EESM 40 (37% RUP) and extruded expeller SBM-EESM 60 (41% RUP) were used in groups IV and V, respectively. Results showed no significant effect on feed intake, crude protein (CP), metabolizable energy (ME), and body condition score (BCS). Ewes fed the 37% RUP diet gained more (p<0.05) weight compared with ewes fed the 31% RUP diet (5.62 vs. 2.5kg). Ewes in EESM 60 had the highest levels of fat, protein, total solid, solid not fat, and immunoglobulin and the lowest in urea N content (P< 0.05) in colostrum during the first 24hrs after lambing. Conclusions: Protein source and RUP levels in ewes’ diets had no significant effect (P< 0.05) on lambs’ birth weight and ewes' blood biochemical parameters. Increasing the RUP content of diet during late gestation resulted in an increase in colostrum constituents and its IgG level but had no effect on ewes’ performance and their lambs’ outcome.

Keywords: colostrum, ewes, lambs output, pregnancy, undegradable protein

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Authors: Shahram Baghban Cirus, Parisa Alizadeh Oskuie

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In a greenhouse experiment, green bean were inoculated with three levels of phosphorus (P1, P2, P3, respectively 0, 50, 100 kgP/h) and four levels of water stress(Fc1, Fc2, Fc3 ,Fc4, respectively 0.8Fc, 0.7Fc, 0.6Fc, 0.5Fc) and one species of VA mycorrhiza (Glomus versiform) or left uninocolated as control plants in the steril soil. AM colonization significantly stimulated plant growth, leaf area, shoot, and pod dry weight but water stress significantly decreased colonization, pod and shoot dry weight, and shoot P. The use P levels significantly increased leaf area, shoot, and pod dry weight, pods length, and colonization.

Keywords: green bean, plant growth, VA mycorrhiza, water-stress

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Authors: Abul Hashem

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The experiment was conducted to find out the effect of different levels of Moringa oleifiera leaf extract and synthetic antioxidant (Beta Hydroxyl Anisole) on fresh and preserved beef meatballs. For this purpose, ground beef samples were divided into five treatment groups. They are treated as control, synthetic antioxidant, 0.1%, 0.2% and 0.3% Moringa oleifera leaf extract as T1, T2, T3, T4 and T5, respectively. Five kinds of meatballs were made and biscuit crushed and egg albumin was mixed with beef meatballs and cooking was practiced properly. Proximate analysis, sensory tests (color, flavor, tenderness, juiciness, overall acceptability), cooking loss, pH value, free fatty acids (FFA), thiobarbituric acid values (TBARS), peroxide value(POV) and microbiological examination were determined in order to evaluate the effect of Moringa oleifiera leaf extract as natural antioxidant & antimicrobial activities in comparing to BHA (Beta Hydroxyl Anisole) at first day before freezing and for maintaining meatballs qualities on the shelf life of beef meat balls stored for 60 days under frozen condition. Freezing temperature was -20˚C. Days of intervals of experiment were on 0, 15th, 30th, and 60th days. Dry matter content of all the treatment groups differ significantly (p<0.05). On the contrary, DM content increased significantly (p<0.05) with the advancement of different days of intervals. CP content of all the treatments were increased significantly (p<0.05) among the different treatment groups. EE content at different treatment levels differ significantly (p<0.05). Ash content at different treatment levels was also differ significantly (p<0.05). FFA values, TBARS, POV were decreased significantly (p<0.05) at different treatment levels. Color, odor, tenderness, juiciness, overall acceptability, raw PH, cooked pH were increased at different treatment levels significantly (p<0.05). The cooking loss (%) at different treatment levels were differ significantly (p<0.05). TVC (logCFU/g), TCC (logCFU/g) and TYMC (logCFU/g) was decreased significantly (p<0.05) at different treatment levels comparison to control. Considering CP, tenderness, juiciness, overall acceptability, cooking loss, FFA, POV, TBARS and microbial parameters it can be concluded that Moringa oleifera leaf extract at 0.1%, 0.2% and 0.3% can be used instead of 0.1% synthetic antioxidant BHA in beef meatballs.

Keywords: antioxidant, beef meatball, BHA, moringa leaf extract, quality

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Authors: N. Mahmoudnia, M. Khormali

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This experiment was conducted to determine the apparent protein digestibility of poultry by- product meal (PBPM) from two industrial poultry slaughter houses on Ross 308 male broiler chickens in independed comparisons. The experiment consisted of seven dietary treatments and three replicates per treatment with three broiler chickens per replicate in a completely randomized design. Dietary treatments consisted of a control corn- soybean diet, and levels 3, 6 and 9% PBPM produced by slaughter house 1 and levels 3, 6 and 9% PBPM produced by slaughter house 2. Chromic oxide was added to the experimental diets as indigestible marker. The apparent protein digestibility of each diet were determined with two methods of sample collection of ileum and excreta in 21-28 d of age. The results this experiment showed that use of PBPM had no significantly effect on performance of broiler chicks during period of experiments. The apparent protein digestibility of PBPM groups was significantly higher than control group by excreta sampling procedure (P<0.05). Using of PBPM 2 significantly (P<0.05) decreased the apparent protein digestibility values based on ileum sampling procedure vs control ( 85.21 vs 90.14).Based results of this experiment,it is possible to use of PBPM 1 in broiler chicken.

Keywords: poultry by-product meal, apparent protein digestibility, independed comparison, broiler chicken

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Authors: Neha Bhadauria, Indu Gaur, Shilpi Shilpi, Susmita Goswami, Prabir K. Paul

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The aerial surface of plants colonized by Human Enteric Pathogens ()has been implicated in outbreaks of enteric diseases in humans. Practice of organic farming primarily using animal dung as manure and sewage water for irrigation are the most significant source of enteric pathogens on the surface of leaves, fruits and vegetables. The present work aims to have an insight into the molecular mechanism of interaction of Human Enteric Pathogens or their metabolites with cell wall receptors in plants. Tomato plants grown under aseptic conditions at 12 hours L/D photoperiod, 25±1°C and 75% RH were inoculated individually with S. fonticola and K. pneumonia. The leaves from treated plants were sampled after 24 and 48 hours of incubation. The cell wall and cytoplasmic proteins were extracted and isocratically separated on 1D SDS-PAGE. The sampled leaves were also subjected to formaldehyde treatment prior to isolation of cytoplasmic proteins to study protein-protein interactions induced by Human Enteric Pathogens. Protein bands extracted from the gel were subjected to MALDI-TOF-TOF MS analysis. The foremost interaction of Human Enteric Pathogens on the plant surface was found to be cell wall bound receptors which possibly set ups a wave a critical protein-protein interaction in cytoplasm. The study revealed the expression and suppression of specific cytoplasmic and cell wall-bound proteins, some of them being important components of signaling pathways. The results also demonstrated HEP induced rearrangement of signaling pathways which possibly are crucial for adaptation of these pathogens to plant surface. At the end of the study, it can be concluded that controlling the over-expression or suppression of these specific proteins rearrange the signaling pathway thus reduces the outbreaks of food-borne illness.

Keywords: cytoplasmic protein, cell wall-bound protein, Human Enteric Pathogen (HEP), protein-protein interaction

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Authors: Arpit Kumar Shrivastava, Subrat Kumar, Rajani Kanta Mohapatra, Priyadarshi Soumyaranjan Sahu

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Computational approaches to predict structure, function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are not effective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical epitopic protein in C. hominis genome through BLASTP analysis. A 3D model of the hypothetical protein was generated using I-Tasser server through threading methodology. The quality of the model was validated through Ramachandran plot by PROCHECK server. The functional annotation of the hypothetical protein through DALI server revealed structural similarity with human Transportin 3. Phylogenetic analysis for this hypothetical protein also showed C. hominis hypothetical protein (CUV04613) was the closely related to human transportin 3 protein. The 3D protein model is further subjected to virtual screening study with inhibitors from the Zinc Database by using Dock Blaster software. Docking study reported N-(3-chlorobenzyl) ethane-1,2-diamine as the best inhibitor in terms of docking score. Docking analysis elucidated that Leu 525, Ile 526, Glu 528, Glu 529 are critical residues for ligand–receptor interactions. The molecular dynamic simulation was done to access the reliability of the binding pose of inhibitor and protein complex using GROMACS software at 10ns time point. Trajectories were analyzed at each 2.5 ns time interval, among which, H-bond with LEU-525 and GLY- 530 are significantly present in MD trajectories. Furthermore, antigenic determinants of the protein were determined with the help of DNA Star software. Our study findings showed a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for control as well as prevention of cryptosporidiosis among humans and animals.

Keywords: cryptosporidium hominis, hypothetical protein, molecular docking, molecular dynamics simulation

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Authors: Mohammad Aladwan

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Alzheimer’s disease (AD) is a widespread dementing illness with a complex and poorly understood etiology. An important role in improving our understanding of the AD process is the modeling of disease-associated changes in tau protein phosphorylation, a protein known to mediate events essential to the onset and progression of AD. A main feature of AD is the abnormal phosphorylation of tau protein and the presence of neurofibrillary tangles. In order to evaluate the respective roles of the microtubule-binding region (MTBR) and alternatively spliced exons in the N-terminal projection domains in AD, we have constructed SHSY-5Y cell lines that stably overexpress four different species of tau protein (4R2N, 4R0N, N(E-2), N(E+2)). Since the toxicity and spreading of tau lesions in AD depends on the interactions of tau with other proteins, we have performed a proteomic analysis of exosome-fraction interactomes for cell lysates and media samples that were isolated from SHSY-5Y cell lines. Functional analysis of tau interactomes based on gene ontology (GO) terms was performed using the String 10.5 database program. The highest number of exosomes proteomes and tau associated proteins were found with 4R2N isoform (2771 and 159) in cell lysate and they have a high strength of connectivity (78%) between proteins, while N(E-2) isoform in the media proteomes has the highest number of proteins and tau associated protein (1829 and 205). Moreover, known AD markers were significantly enriched in secreted interactomes relative to lysate interactomes in the SHSY-5Y cells of tau isoforms lacking exons 2 and 3 in the N-terminal. The lack of exon 2 (E-2) from tau protein can be mediated by tau secretion and spreading to different cells. Enriched functions in the secreted E-2 interactome include signaling and developmental pathways that have been linked to a) tau misprocessing and lesion development and b) tau secretion and which, therefore, could play novel roles in AD pathogenesis.

Keywords: Alzheimer's disease, dementia, tau protein, neurodegenration disease

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Authors: Homayun Ghasemi, Mojtaba Yousefirad, Mozhgan Farzamisepehr

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Heavy metals are among soil pollutant resources that in case of accumulation in the soil and absorption by the plant, enter into the food chain and poison the plants or the people who consume those plants. This research was performed in order to examine the role of cadmium as a heavy metal in the activity of catalase and peroxidase as well as protein concentration in Trifolium resupinatum L. based on a randomized block design with three repetitions. The used treatments included consumption of Cd (NO3)2 at four levels, namely, 0, 100, 200, and 300 ppm. The plants under study were treated for 10 days. The results of the study showed that catalase activity decreased by the increase of cadmium. Moreover, peroxidase activity increased by an increase inthe consumption of cadmium. The analysis of protein level showed that plantlet protein decreased in high cadmium concentrations. The findings also demonstrated that cadmium concentration in roots was higher than in shoots.

Keywords: catalase, heavy metal, peroxidase, protein

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Authors: Victor Prevost, Solene Landerneau, Michel Duhamel, Joel Sternheimer, Olivier Gallet, Pedro Ferrandiz, Marwa Mokni

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The search for homologous proteins is one of the ongoing challenges in biology and bioinformatics. Traditionally, a pair of proteins is thought to be homologous when they originate from the same ancestral protein. In such a case, their sequences share similarities, and advanced scientific research effort is spent to investigate this question. On this basis, we propose the Protein-Wave Alignment Tool (”P-WAT”) developed within the framework of the France Relance 2030 plan. Our work takes into consideration the mass-related wave aspect of protein biosynthesis, by associating specific frequencies to each amino acid according to its mass. Amino acids are then regrouped within their mass category. This way, our algorithm produces specific alignments in addition to those obtained with a common amino acid coding system. For this purpose, we develop the ”P-WAT” original algorithm, able to address large protein databases, with different attributes such as species, protein names, etc. that allow us to align user’s requests with a set of specific protein sequences. The primary intent of this algorithm is to achieve efficient alignments, in this specific conceptual frame, by minimizing execution costs and information loss. Our algorithm identifies sequence similarities by searching for matches of sub-sequences of different sizes, referred to as primers. Our algorithm relies on Boolean operations upon a dot plot matrix to identify primer amino acids common to both proteins which are likely to be part of a significant alignment of peptides. From those primers, dynamic programming-like traceback operations generate alignments and alignment scores based on an adjusted PAM250 matrix.

Keywords: protein, alignment, homologous, Genodic

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Authors: Sulyman Abdulhakeem Olarewaju, S. O. Malomo, M. T. Yakubu, J. O. Akolade

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The ameliorative effect of Celosia argentea var. cristata leaf extract against cadmium (Cd) induced oxidative stress and toxicity in selected tissues of rats was investigated. Toxicity coupled with oxidative stress was induced in rats by oral administration of Cd (8 mg/kg b. wt). Preliminary quantitative phytochemical and in vitro antioxidant analyses showed that the methanolic extract of C. argentea leaves was constituted by polyphenols (5.72%), saponins (3.20%), tannins (0.65%) and cadenolides (0.006%). IC50 of 9800, 7406, and 45.04 μg/ml were recorded for inhibition of linoleic acid oxidation, 2, 2-diphenyl-1-picrylhydrazyl and hydrogen peroxide radicals respectively. Simultaneous administration of C. argentea leaf extract with Cd significantly attenuated Cd-induced elevation of serum enzyme markers such as aspartate and alanine transaminase, alkaline and acid phosphatase as well as γ-glutaryltransferase in a dose-dependent fashion, while their reduced level in the liver were significantly increased. Higher levels of enzymatic antioxidants; superoxide dismutase and catalase activities were observed in the liver, brain, kidney and testes of the Cd-induced rats treated with C. argentea extract, while lipid peroxidation expressed in malondialdehyde concentrations were lower when compared to values in rats administered Cd only. Other Cd-induced toxicity and stress markers in the serum viz. reduced uric acid and albumin levels as well as elevated total and unconjugated bilirubin were attenuated by the extract and their values compared favorably with those animals co-administered cadmium with ascorbic acid. Data from the study showed that oral administration of extract from the leaf C. argentea may ameliorate Cd-induced oxidative stress and toxicity in rats.

Keywords: toxicity, cadmium, celosia, antioxidants, oxidative stress

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Authors: Mukesh Saran, Ashima Bagaria

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Green nanotechnology is the most researched field in the current scenario. Herein we study the synthesis of Zinc and Ferrous nanoparticles using Moringa oleifera leaf extracts. Our protocol using established protocols heat treatment of plant extracts along with the solution of copper sulphate in the ratio of 1:1. The leaf extracts of Moringa oleifera were prepared in deionized water. Copper sulfate solution (1mM) was added to this, and the change in color of the solution was observed indicating the formation of Cu nanoparticles. The as biosynthesized Cu nanoparticles were characterized with the help of Scanning Electron Microscopy (SEM), and Fourier Transforms Infrared Spectroscopy (FTIR). It was observed that the leaf extracts of Moringa oleifera can reduce copper ions into copper nanoparticles within 8 to 10 min of reaction time. The method thus can be used for rapid and eco-friendly biosynthesis of stable copper nanoparticles. Further, we checked their antimicrobial and antioxidant potential, and it was observed that maximum antioxidant activity was observed for the particles prepared using the heating method. The maximum antibacterial activity was observed in Streptomyces grisveus particles and in Triochoderma Reesei for the maximum antifungal activity. At present, we are engaged in studying the anti-inflammatory activities of these as prepared nanoparticles.

Keywords: green synthesis, antibacterial, antioxidant, antifungal, anti-inflammatory

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Authors: Friday A. Ogori, Ojotu M. Eke, Oneh J. Abu, Abraham T. Girgih

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B.aeqygtiaca del (Balanites aegyptiaca del) seed protein concentrate (APC) was hydrolyzed using different enzymes such as pepsin+pancreatin (PP), Alcalase (Alca), and Flavourzyme (Flav). The Alca has higher yield (100%) when compared to PP (83.23%) and Flav (62.90%). The hydrophobic amino acid and Sulphur containing amino acid (40.19%, 7.04%) in PP hydrolysate were higher compared to Alcalase (38.92%, 6.69%), Flavourenzyme (37.43%,6.35%), and APC (39.97%, 6.95%) samples. The PP has stronger DPPH, Hydroxyl radical quenching, Ferric reducing activity, and linoleic acid peroxidation activity, followed by the protein concentrate (APC) and Alcalase (Alca), while Flavourenzyme (Flav) derived hydrolysate was least in scavenging and inhibiting radical peroxidation properties. Flavourenzyme derived hydrolysate had stronger Ferric reducing antioxidant potential and metal chelating property. The result showed that the Alcalase hydrolysate has promising peptide yield, and PP hydrolysate had excellent amino acid residues and good in-vitro antioxidant potentials and could be a preferred ingredients in the nutraceutical and functional food emerging industries.

Keywords: balanites aegyptiaca del, protein concentrate, protein hydrolysates, enzymatic hydrolysis, antioxidants

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Authors: Thanusha D. Abeywickrama, Inoka C. Perera, Genji Kurisu

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Tuberculosis, considered being as the ninth leading cause of death worldwide, cause from a single infectious agent M. tuberculosis and the drug resistance nature of this bacterium is a continuing threat to the world. Therefore TB preventing treatment is expanding, where this study designed to analyze the regulatory mechanism of GntR transcriptional regulator gene Rv0792c, which lie between several genes codes for some hypothetical proteins, a monooxygenase and an oxidoreductase. The gene encoding Rv0792c was cloned into pET28a and expressed protein was purified to near homogeneity by Nickel affinity chromatography. It was previously reported that the protein binds within the intergenic region (BS region) between Rv0792c gene and monooxygenase (Rv0793). This resulted in binding of three protein molecules with the BS region suggesting tight control of monooxygenase as well as its own gene. Since monooxygenase plays a key role in metabolism, this gene may have a global regulatory role. The natural ligand for this regulator is still under investigation. In relation to the Rv0792 protein structure, a Circular Dichroism (CD) spectrum was carried out to determine its secondary structure elements. Percentage-wise, 17.4% Helix, 21.8% Antiparallel, 5.1% Parallel, 12.3% turn and 43.5% other were revealed from CD spectrum data under room temperature. Differential Scanning Calorimetry (DSC) was conducted to assess the thermal stability of Rv0792, which the melting temperature of protein is 57.2 ± 0.6 °C. The graph of heat capacity (Cp) versus temperature for the best fit was obtained for non-two-state model, which concludes the folding of Rv0792 protein occurs through stable intermediates. Peak area (∆HCal ) and Peak shape (∆HVant ) was calculated from the graph and ∆HCal / ∆HVant was close to 0.5, suggesting dimeric nature of the protein.

Keywords: CD spectrum, DSC analysis, GntR transcriptional regulator, protein structure

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Authors: Jaturong Matidtor, Krisna R. Torrissen, Saengtong Pongjareankit, Sudaporn Tongsiri, Jiraporn Rojtinnakorn

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Low survival rate has being particular problem in nursery of marble goby juvenile. The aim of this study was to investigate effect of garlic extract on protein digestive pancreatic enzymes, trypsin (T) and chymotrypsin (C). The marble goby were reared with commercial feed mixed garlic extract at concentration of 0 (control), 0.3, 0.5, 1.0, 3.0 and 5.0% (w/w) for 6 weeks. Analysis of the digestive enzymes at 2 and 6 weeks was performed. Growth parameters; weight gain (WG), specific growth rate (SGR) and feed efficiency (FE), were identified. For T, C and T/C at 2 weeks, values of T and T/C ratio of 0.3% (w/w) group showed significant difference (p < 0.05) with the highest values of 17685.64± 11981.77 U/mg protein and of 51.64 ± 27.46 U/mg protein, respectively. For C at 2 weeks, 0% (w/w) group showed the highest values of 16191.76± 2225.56 U/mg protein. Whereas value of T, C and T/C ratio at 6 weeks, there was no significant difference (p > 0.05). For growth performance, it significantly increased in all garlic extract fed groups (0.3-5.0%, w/w), both at 2 and 6 weeks. At 2 weeks, values of WG and SGR of 0.5% (w/w) group showed the highest values of 71.51 ± 1.60%, and 3.85 ± 0.07%, respectively. For FE, 0.3% (w/w) group showed the highest value of 60.21 ± 6.51%. At 6 weeks, it illustrated that all growth parameters of 5.0% (w/w) group were the highest values; WG = 35.06 ± 5.66%, SGR = 2.14 ± 0.30%, and FE = 5.86 ± 0.68%. We suggested that garlic extract could be available for protein digestive enzyme and growth enhancement in marble goby nursery with artificial feed. This result will be high benefit for commercial aquaculture of marble goby.

Keywords: marble goby, nursery, garlic extract, digestive enzyme, growth

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Authors: Rizky Kusuma Cahyani

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Hepatitis B is a liver inflammatory disease caused by hepatitis B virus (HBV). This infection can be prevented by vaccination which contains HBV surface protein (sHBsAg). However, vaccine supply is limited. Several attempts have been conducted to produce local sHBsAg. However, the purity degree and protein yield are still inadequate. Therefore optimization of HBsAg purification steps is required to obtain high yield with better purification fold. In this study, optimization of purification was done in 2 steps, precipitation using variation of NaCl concentration (0,3 M; 0,5 M; 0,7 M) and PEG (3%, 5%, 7%); ion exchange chromatography (IEC) using NaCl 300-500 mM elution buffer concentration.To determine HBsAg protein, bicinchoninic acid assay (BCA) and enzyme-linked immunosorbent assay (ELISA) was used in this study. Visualization of HBsAg protein was done by SDS-PAGE analysis. Based on quantitative analysis, optimal condition at precipitation step was given 0,3 M NaCl and PEG 3%, while in ion exchange chromatography step, the optimum condition when protein eluted with NaCl 500 mM. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis indicates that the presence of protein HBsAg with a molecular weight of 25 kDa (monomer) and 50 kDa (dimer). The optimum condition for purification of sHBsAg produced in Pichia pastoris gave a yield of 47% and purification fold 17x so that it would increase the production of hepatitis B vaccine to be more optimal.

Keywords: hepatitis B virus, HBsAg, hepatitis B surface antigen, Pichia pastoris, purification

Procedia PDF Downloads 151

Authors: S. Bilal, S. A. Bhat, I. Hussain, J. D. Parrah, S. P. Ahmad, M. R. Mir

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Background: The wound healing involves a highly coordinated cascade of cellular and immunological response over a period including coagulation, inflammation, granulation tissue formation, epithelialization, collagen synthesis and tissue remodeling. Wounds in aged heal more slowly than those in younger, mainly because of comorbidities that occur as one age. The present study is about the influence of age on wound healing. 1x1cm^2 (100 mm) wounds were created on the back of the animal. The animals were divided into two groups; one group had animals in the age group of 3-9 months while another group had animals in the age group of 15-21 months. Materials and Methods: 24 clinically healthy rabbits in the age group of 3-21 months were used as experimental animals and divided into two groups viz A and B. All experimental parameters, i.e., Excision wound model, Measurement of wound area, Protein extraction and estimation, Protein extraction and estimation and DNA extraction and estimation were done by standard methods. Results: The parameters studied were wound contraction, hydroxyproline, glucosamine, protein, and DNA. A significant increase (p<0.005) in the hydroxyproline, glucosamine, protein and DNA and a significant decrease in wound area (p<0.005) was observed in the age group of 3-9 months when compared to animals of an age group of 15-21 months. Wound contraction together with hydroxyproline, glucosamine, protein and DNA estimations suggest that advanced age results in retarded wound healing. Conclusion: The decrease wound contraction and accumulation of hydroxyproline, glucosamine, protein and DNA in group B animals may be associated with the reduction or delay in growth factors because of the advancing age.

Keywords: age, wound healing, excision wound, hydroxyproline, glucosamine

Procedia PDF Downloads 659

Authors: Şevket Evci, Mehmet Akif Karsli

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The aim of this study is to determine nutrient contents, in situ ruminal degradation kinetics and protein fractions of screenings bean (B), chick pea (ChP), red lentil (RL) and green lentil (GL) that is used as residue in grain legume packing industry. For this purpose, four samples of each legumes species-a total of 16 samples, collected from different parts of our country were utilized. Feedstuffs used in the experiment were incubated for 0, 2 4, 8, 12, 24, and 48 hours in the rumen of 3 ruminally cannulated Akkaraman rams as duplicate. The nutrient contents, in situ ruminal dry matter (DM), organic matter (OM) and crude protein (CP) degradabilities and fractions, and escape protein contents were evaluated. The highest OM and CP contents were observed in RL (P<0.05). Chick pea had the highest ether extract (EE) content and EE values were 3.47, 6.72, 2.26, 8.66 % for RL, B, GL and ChP, respectively (P<0.05). Crude fiber (CF), ADF, and NDF contents were the highest in RL and the lowest in ChP. CF values were 24.03, 10.80, 4.09 and 3.57 % for RL, GL, B and ChP (P<0.05). Acid detergent insoluble nitrogen content of samples did not differ. Escape protein content was the highest in RL and the lowest in B (P<0.05). After 48 h incubation, the lowest OM and CP degradabilities were observed in RL. While the highest OM degradability was seen in ChP the highest CP degradability was observed in B (P<0.05). The lowest water soluble OM and CP contents were observed in RL whereas the highest potentially degradable OM and CP contents were seen in B and ChP (P<0.05). Both rate of OM and CP degradations (k-1) did not differ among samples (P>0.05). In conclusion, it was noted that feedstuffs (GL, ChP and B) used in the experiment except RL had a greater ruminal degradibilities of both OM and CP and moreover, had a higher escape protein contents, except B. It was thought that these feedstuffs can be substituted with some of common protein sources used in animal nutrition.

Keywords: in situ, nutrient contents, ruminant, subsieve

Procedia PDF Downloads 481

Authors: Lavinia Ruta, Ileana Farcasanu

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The yeast surface display (YSD) technique enables the expression of proteins on yeast cell surfaces, facilitating the identification and isolation of proteins with targeted binding properties, such as nanobodies. Nanobodies, derived from camelid species, are single-domain antibody fragments renowned for their high affinity and specificity towards target proteins, making them valuable in research and potentially in therapeutics. Their advantages include a compact size (~15 kDa), robust stability, and the ability to target challenging epitopes. The project endeavors to establish and validate a platform for producing Green Fluorescent Protein (GFP) and mCherry nanobodies using the yeast surface display method. mCherry, a prevalent red fluorescent protein sourced from coral species, is commonly utilized as a genetic marker in biological studies due to its vibrant red fluorescence. The GFP-nanobody, a single variable domain of heavy-chain antibodies (VHH), exhibits specific binding to GFP, offering a potent means for isolating and engineering fluorescent protein fusions across various biological research domains. Both GFP and mCherry nanobodies find specific utility in cellular imaging and protein analysis applications.

Keywords: YSD, nanobodies, GFP, Saccharomyces cerevisiae

Procedia PDF Downloads 61

Authors: Vineeta Kaushik, Manisha Goel

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Chaperones are proteins that help other proteins fold correctly, and are found in all domains of life i.e., prokaryotes, eukaryotes and archaea. Various comparative genomic studies have suggested that the archaeal protein folding machinery appears to be highly similar to that found in eukaryotes. In case of protein folding; slow rotation of peptide prolyl-imide bond is often the rate limiting step. Formation of the prolyl-imide bond during the folding of a protein requires the assistance of other proteins, termed as peptide prolyl cis-trans isomerases (PPIases). Cyclophilins constitute the class of peptide prolyl isomerases with a wide range of biological function like protein folding, signaling and chaperoning. Most of the cyclophilins exhibit PPIase enzymatic activity and play active role in substrate protein folding which classifies them as a category of molecular chaperones. Till date, there is not very much data available in the literature on archaeal cyclophilins. We aim to compare the structural and biochemical features of the cyclophilin protein from within the three domains to elucidate the features affecting their stability and enzyme activity. In the present study, we carry out in-silico analysis of the cyclophilin proteins to predict their conserved residues, sites under positive selection and compare these proteins to their bacterial and eukaryotic counterparts to predict functional divergence. We also aim to clone and express these proteins in heterologous system and study their biophysical characteristics in detail using techniques like CD and fluorescence spectroscopy. Overall we aim to understand the features contributing to the folding, stability and dynamics of the archaeal cyclophilin proteins.

Keywords: biophysical characterization, x-ray crystallography, chaperone-like activity, cyclophilin, PPIase activity

Procedia PDF Downloads 213

Authors: Sosina Adedayo O., Babyemi Olaniyi J.

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The location, population, and distribution pattern of sheep are severe challenges to agribusiness investment and policy formulation in the livestock industry. There is a significant disconnect between farmers' needs and the policy framework towards ameliorating the sheep production constraints. Information on the population, production, and distribution pattern of sheep remains very scanty. A multi-stage sampling technique was used to elicit information from 180 purposively selected respondents from the study area comprised of Oluyole, Ona-ara, Akinyele, Egbeda, Ido and Ibarapa East LGA. The Global Positioning Systems (GPS) of the farmers' location (distribution), and average sheep herd size (Total Livestock Unit, TLU) (population) were recorded, taking the longitude and latitude of the locations in question. The recorded GPS data of the study area were transferred into the ARC-GIS. The ARC-GIS software processed the data using the ARC-GIS model 10.0. Sheep production and distribution (TLU) ranged from 4.1 (Oluyole) to 25.0 (Ibarapa East), with Oluyole, Akinyele, Ona-ara and Egbeda having TLU of 5, 7, 8 and 20, respectively. The herd sizes were classified as less than 8 (smallholders), 9-25 (medium), 26-50 (large), and above 50 (commercial). The majority (45%) of farmers were smallholders. The FR CP (%) ranged from 5.81±0.26 (cassava leaf) to 24.91±0.91 (Amaranthus spinosus), NDF (%) ranged from 22.38±4.43 (Amaranthus spinosus) to 67.96 ± 2.58 (Althemanthe dedentata) while ME ranged from 7.88±0.24 (Althemanthe dedentata) to 10.68±0.18 (cassava leaf). The smallholders’ sheep farmers were the majority, evenly distributed across rural areas due to the availability of abundant feed resources (crop residues, tree crops, shrubs, natural pastures, and feed ingredients) coupled with a large expanse of land in the study area. Most feed resources available were below sheep protein requirement level, hence supplementation is necessary for productivity. Bio-informatics can provide relevant information for sheep production for policy framework and intervention strategies.

Keywords: sheep enterprise, agribusiness investment, policy, bio-informatics, ecological zone

Procedia PDF Downloads 82

Authors: Meenakshi Verma, Mandeep Singh Bakshi, Kultar Singh

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Magnetic nanoparticles have received incredible importance in view of their diverse applications, which arise primarily due to their response to the external magnetic field. The magnetic behaviour of magnetic nanoparticles (NPs) helps them in numerous different ways. The most important amongst them is the ease with which they can be purified and also can be separated from the media in which they are present merely by applying an external magnetic field. This exceptional ease of separation of the magnetic NPs from an aqueous media enables them to use for extracting/removing metal pollutants from complex aqueous medium. Functionalized magnetic NPs can be subjected for the metallic impurities extraction if are favourably adsorbed on the NPs surfaces. We have successfully used the magnetic NPs as vehicles for gold and silver NPs removal from the complex fluids. The NPs loaded with gold and silver NPs pollutant fractions has been easily removed from the aqueous media by using external magnetic field. Similarly, we have used the magnetic NPs for extraction of protein from complex media and then constantly washed with pure water to eliminate the unwanted surface adsorbed components for quantitative estimation. The purified and protein loaded magnetic NPs are best analyzed with SDS Page to not only for characterization but also for separating the protein fractions. A collective review of the results indicates that we have synthesized surfactant coated iron oxide NPs and then functionalized these with selected materials. These surface active magnetic NPs work very well for the extraction of metallic NPs from the aqueous bulk and make the whole process environmentally sustainable. Also, magnetic NPs-Au/Ag/Pd hybrids have excellent protein extracting properties. They are much easier to use in order to extract the magnetic impurities as well as protein fractions under the effect of external magnetic field without any complex conventional purification methods.

Keywords: magnetic nanoparticles, protein, functionalized, extraction

Procedia PDF Downloads 99