Search results for: protein fractions

Authors: Chaoran Liu

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Bioactive polysaccharides and polysaccharide-protein complex derived from mushrooms and fungi have a wide range of immunomodulatory activity with low side-effects and have therefore the potential to be developed as an adjuvant in cancer therapies. Mushrooms sclerotium is rich in polysaccharides and the polysaccharides isolated from the sclerotium of Polyporus rhinocerus have shown potent in vivo and in vitro immunomodulatory effects. Macrophages are considered to be an important component of the innate immune response against bacterial infection and cancer. To better understanding the immunomodulatory effects and its underlying mechanisms of sclerotial water-soluble polysaccharides extracted from P. rhinocerus on macrophages, the objectives of this study are to purify the water-soluble novel sclerotial polysaccharides and to characterize the structure and properties as well as to study the detailed molecular mechanisms of the in vitro immunomodulating effects in murine macrophages. The hot water-soluble fraction PRW from the sclerotium of P. rhinocerus was obtained using solvent extraction. PRW was further fractionated by membrane ultrafiltration to a give a fraction (PRW1) with molecular mass less than 50 kDa. PRW1 was characterized to be a polysaccharide-protein complex composed of 45.7% polysaccharide and 44.2% protein. The chemical structure of the carbohydrate moiety of PRW1 was elucidated by GC and FTIR to be mainly beta-D-glucan with trace amount of galactose and mannose. The immunomodulatory effects of PRW1 on murine RAW 264.7 macrophages were demonstrated in terms of the increase in nitric oxide production and cytokine production. Mechanistically, PRW1 initiates ERK phosphorylation to activate macrophages within 15 min and significantly improves the expression level of inducible NOS (iNOS) from 6 h after treatment. In summary, this study indicates that PRW1 is a potent immunomodulatory agent for macrophages and suggests that mushroom sclerotia from Polyporus rhinocerus requires for further investigation in cancer research.

Keywords: Polyporus rhinocerus, mushroom sclerotia, Polysaccharide-Protein Complex, macrophage activation

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Authors: Tagouelbe Tiho, Amissa Augustin Adima, Yao Casimir Brou, Nabayo Traore, Gouha Firmin Kouassi, Thierry Roland Kouame, Maryline Kouba

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The work aimed to study Borassus aethiopum Mart (B.a) dried pulp nutritional value for its incorporation in human and poultry diets. Firstly, the mature fruit macro-composition was assessed. Secondly, the pulp was dried at 40, 50, 60, 70, and 80ᵒC. Thereafter, the analysis was performed for fat, protein, total sugars, Ca, P, Mg, and fatty acid profile monitoring. As a result, the fruits weighed 1,591.35, delivered 516.73, and 677.82 grams of pulp and seeds, respectively. Mainly, increasing heat adversely affected the outputs. Consequently, the fat results were 14.12, 12.97, 8.93, 8.89ᶜ, and 5.56%; protein contents were 11.64, 10.15, 8.97, 8.84, and 8.42%; total sugar deliveries were 6.28, 6.05, 5.26, 5.02, and 4.76% (P < 0.01). Thereafter, the metabolizable energies were 3,785.22; 3,834.28; 3,616.62; 3,667.03; and 3,608.33 kcal/kg (DM). Additionally, Calcium (Ca) contents were 0.51, 0.55, 0.69, 0.77, and 0.81%, while phosphorus (P) mean was 0.17%, and the differences were not significant (P < 0.01). So, the Ca/P ratios were 2.79, 3.04, 4.10, 4.71, and 4.95. Finally, fatty acids (FA) assessments revealed 22.33 saturated (SFA), 77.67 unsaturated (UFA), within which 67.59% were monounsaturated (MUFA). Interestingly, the rising heat depressed n-6/n-3 ratios that were 1.1, 1.1, 0.45 and 0.38, respectively at 40, 50, 70 and 80ᵒC. In short, drying did not only enhance the product shelf life but it also improved the nutritional value. Thus, B.a mature fruit pulps dried at 70ᵒC are good functional foods, with more than 66% MUFA, and energy source for human and poultry nutrition.

Keywords: Borassus aethiopum Mart, fatty acids, metabolizable energy, minerals, protein

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Authors: Hamad Rafique

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The bioactive peptide RDFPITWPW (RW-9) identified from oat protein has been reported to be positive in memory deficits. However, no clarity on the mechanisms responsible for the neuroprotective effects of RW-9 peptide against AD-like symptoms. Herein, it found that RW-9 intervention showed various improving effects in cognitive-behavioral tests and alleviated oxidative stress and inflammation in the scopolamine-induced mice model. The hippocampus proteomics analysis revealed the upregulation of memory-related proteins, including Grin3a, Ppp2r1b, Stat6, Pik3cd, Slc5a7, Chrm2, mainly involved in cAMP signaling, PI3K-Akt signaling, and JAK-STAT signaling pathways. The administration of RW-9 significantly upregulated the neurotransmitters, including 5-HT, DA, and Arg, in mice brains. Moreover, it regulated the serum metabolic profile and increased the expression levels of ABC transporters, biosynthesis of amino acids, and Amino acyl-tRNA biosynthesis, among others. The 16s-rRNA results illustrated that the RW-9 restored the abundance of Muribaculaceae, Lachnospiraceae, Lactobacillus, Clostridia and Bactericides. Taken together, our results suggest that the RW-9 may prevent the AD-like symptoms via modulation of the gut-serum-brain axis.

Keywords: oat protein, active peptide, neuroprotective, gut-brain axis

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Authors: M. Michael Gromiha, A. Mary Thangakani, Sandeep Kumar, D. Velmurugan

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The main cause of several neurodegenerative diseases such as Alzhemier, Parkinson, and spongiform encephalopathies is formation of amyloid fibrils and plaques in proteins. We have analyzed different sets of proteins and peptides to understand the influence of sequence-based features on protein aggregation process. The comparison of 373 pairs of homologous mesophilic and thermophilic proteins showed that aggregation-prone regions (APRs) are present in both. But, the thermophilic protein monomers show greater ability to ‘stow away’ the APRs in their hydrophobic cores and protect them from solvent exposure. The comparison of amyloid forming and amorphous b-aggregating hexapeptides suggested distinct preferences for specific residues at the six positions as well as all possible combinations of nine residue pairs. The compositions of residues at different positions and residue pairs have been converted into energy potentials and utilized for distinguishing between amyloid forming and amorphous b-aggregating peptides. Our method could correctly identify the amyloid forming peptides at an accuracy of 95-100% in different datasets of peptides.

Keywords: aggregation, amyloids, thermophilic proteins, amino acid residues, machine learning techniques

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Authors: Natela Gerliani, Mohammed Aider

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Nowadays, the interest of using sustainable technologies for protein extraction from underutilized oilseeds is growing. Currently, a major disposal problem for the oil industry is by-products of plant food processing such as soybean meal. That is why valorization of soybean meal is important for the oil industry since it contains high-quality proteins and other valuable components. Generally, soybean meal is used in livestock and poultry feed but is rarely used in human feed. Though chemical composition of this meal compensate nutritional deficiency and can be used to balance protein in human food. Regarding the efficiency of soybean meal valorization, extraction is a key process for obtaining enriched protein ingredient, which can be incorporated into the food matrix. However, most of the food components such as proteins extracted from oilseeds by-products imply the utilization of organic and inorganic chemicals (e.g. acids, bases, TCA-acetone) having a significant environmental impact. In a context of sustainable production, the use of an electro-activation technology seems to be a good alternative. Indeed, the electro-activation technology requires only water, food grade salt and electricity as main materials. Moreover, this innovative technology helps to avoid special equipment and trainings for workers safety as well as transport and storage of hazardous materials. Electro-activation is a technology based on applied electrochemistry for the generation of acidic and alkaline solutions on the basis of the oxidation-reduction reactions that occur at the vicinity electrode/solution interfaces. It is an eco-friendly process that can be used to replace the conventional acidic and alkaline extraction. In this research, the electro-activation technology for protein extraction from soybean meal was carried out in the electro-activation reactor. This reactor consists of three compartments separated by cation and anion exchange membranes that allow creating non-contacting acidic and basic solutions. Different current intensities (150 mA, 300 mA and 450 mA) and treatment durations (10 min, 30 min and 50 min) were tested. The results showed that the extracts obtained by the electro-activation method have good quality in comparison to conventional extracts. For instance, extractability obtained with electro-activation method was 55% whereas with the conventional method it was only 36%. Moreover, a maximum protein quantity of 48 % in the extract was obtained with the electro-activation technology comparing to the maximum amount of protein obtained by conventional extraction of 41 %. Hence, the environmentally sustainable electro-activation technology seems to be a promising type of protein extraction that can replace conventional extraction technology.

Keywords: by-products, eco-friendly technology, electro-activation, soybean meal

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Authors: Abdu Umar Adamu, Saidu Akun Abdullahi, Al-Hassan Muhammed, Hamisu Abdu

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Leguminous plants such as beans have been considered as a source of protein in this present work. The proximate analysis on beans (Vigna unguiculata) were determined in order to identify the nutritional content as well as presence of some heavy metals accumulation in washed and unwashed beans (white Beans) sold in Kazaure market Jigawa State Nigeria. On the average comparative analysis, the result has indicated that, the Vigna unguiculata had protein content of 61.1%, fibre 4.5%, ash 10.4%, moisture 5%, carbohydrate 15.8% and total lipid 4.9%, therefore it could be suggested that beans has enough nutritional content that helps the people health. The heavy metal analysis of unwashed white beans showed that Fe (17.37 ± 6.71)mg/kg had the highest concentration followed by Zn (6.41 ± 3.09), Cu (5.69 ± 2.42), Cd (0.46 ± 0.65) and Pb (0.57 ± 0.94)mg/kg , while the washed beans shows that Zn (0.11 ± 0.17), Fe (0.01 ± 0.006), Cd (0.02 ± 0.01), Cu (0.03 ± 0.021), Pb (0.01 ± 0.006)mg/kg. The washed white beans are safe for consumption and also the concentration of heavy metal are negligible and of nontoxic effect to human health.

Keywords: white beans, protein, proximate composition, heavy metal

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Authors: A. Saghatelyan, Sh. Asmaryan, G. Tepanosyan, V. Muradyan

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One of major concern to Armenia has been soil degradation emerged as a result of unsustainable management and use of grasslands, this in turn largely impacting environment, agriculture and finally human health. Hence, assessment of soil degradation is an essential and urgent objective set out to measure its possible consequences and develop a potential management strategy. Since recently, an essential tool for assessing pasture degradation has been remote sensing (RS) technologies. This research was done with an intention to measure preciseness of Linear spectral unmixing (LSU) and NDVI-SMA methods to estimate soil surface components related to degradation (fractional vegetation cover-FVC, bare soils fractions, surface rock cover) and determine appropriateness of these methods for mapping man-induced soil degradation in high mountain pastures. Taking into consideration a spatially complex and heterogeneous biogeophysical structure of the studied site, we used high resolution multispectral QuickBird imagery of a pasture site in one of Armenia’s rural communities - Nerkin Sasoonashen. The accuracy assessment was done by comparing between the land cover abundance data derived through RS methods and the ground truth land cover abundance data. A significant regression was established between ground truth FVC estimate and both NDVI-LSU and LSU - produced vegetation abundance data (R2=0.636, R2=0.625, respectively). For bare soil fractions linear regression produced a general coefficient of determination R2=0.708. Because of poor spectral resolution of the QuickBird imagery LSU failed with assessment of surface rock abundance (R2=0.015). It has been well documented by this particular research, that reduction in vegetation cover runs in parallel with increase in man-induced soil degradation, whereas in the absence of man-induced soil degradation a bare soil fraction does not exceed a certain level. The outcomes show that the proposed method of man-induced soil degradation assessment through FVC, bare soil fractions and field data adequately reflects the current status of soil degradation throughout the studied pasture site and may be employed as an alternate of more complicated models for soil degradation assessment.

Keywords: Armenia, linear spectral unmixing, remote sensing, soil degradation

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Authors: Ruolan Zhou

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Trisomy of human chromosome 21 is the primary cause of Down Syndrome (DS), and this genetic disease has significantly burdened families and countries, causing great controversy. To address this problem, the research takes an approach in exploring the relationship between genetic abnormality and this disease's symptoms, adopting several techniques, including data analysis and enrichment analysis. It also explores open-source websites, such as NCBI, DAVID, SOURCE, STRING, as well as UCSC, to complement its result. This research has analyzed the variety of genes on human chromosome 21 with simple coding, and by using analysis, it has specified the protein-coding genes, their function, and their location. By using enrichment analysis, this paper has found the abundance of keratin production-related coding-proteins on human chromosome 21. By adopting past researches, this research has attempted to disclose the relationship between trisomy of human chromosome 21 and keratin production abnormality, which might be the reason for common diseases in patients with Down Syndrome. At last, by addressing the advantage and insufficiency of this research, the discussion has provided specific directions for future research.

Keywords: Down Syndrome, protein production, genome, enrichment analysis

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Authors: Anjum-Zubair, Muhammad, Muhammad Shoaib Alam, Muhammad Samee Mubarik, Iftikhar Ahmad

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The aim of present research was to evaluate the effect of dietary protein (soybean) formulated feed on the growth performance of carp fish seed (Rohu, Mori, Grass, and Gulfam) in ponds under polyculture system. Keeping in view the protein requirements of these four carps, they were fed with formulated feed contains 30% of crude protein. The fingerlings were fed once on daily basis at 5% of their wet body weight. A 90 days experiment was conducted in two cemented ponds situated at Fish Seed Hatchery and Research Centre, Rawal Town, Islamabad, Pakistan. Pond1 contain major carps i.e. Rohu and Mori while pond 2 was stocked with Chinese carps i.e. Grass carp and Gulfam. Random sampling of five individuals of each species was done fortnightly to measure the body weight and total body length. Maximum growth was observed in fingerling of Grass carp followed by Mori, Rohu and Gulfam. Total fish production was recorded as Grass 623.45 gm followed by Mori 260.3 gm, Rohu 243.08 gm and Gulfam 181.165 gm respectively. Significantly results were obtained among these four fish species when the corresponding data was subjected to statistical analysis by using two sample t-test. The survival rate was 100%. Study shows that soybean as plant based protein can be easily used as substitute to fish meal without any adverse effect on fish health and fish production.

Keywords: carps, fry growth, poly culture, soybean meal

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Authors: Neha Babbar, S. Van Roy, W. Dejonghe, S. Sforza, K. Elst

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Pectic oligosaccharides (a class of prebiotics) are non-digestible carbohydrates which benefits the host by stimulating the growth of healthy gut micro flora. Production of prebiotic pectic oligosaccharides (POS) from pectin rich agricultural residues involves a cutting of long chain polymer of pectin to oligomers of pectin while avoiding the formation of monosaccharides. The objective of the present study is to develop a two-step continuous biocatalytic membrane reactor (MER) for the continuous production of POS (from sugar beet pulp) in which conversion is combined with separation. Optimization of the ratio of POS/monosaccharides, stability and productivities of the process was done by testing various residence times (RT) in the reactor vessel with diluted (10 RT, 20 RT, and 30 RT) and undiluted (30 RT, 40 RT and 60 RT) substrate. The results show that the most stable processes (steady state) were 20 RT and 30 RT for diluted substrate and 40 RT and 60 RT for undiluted substrate. The highest volumetric and specific productivities of 20 g/L/h and 11 g/gE/h; 17 g/l/h and 9 g/gE/h were respectively obtained with 20 RT (diluted substrate) and 40 RT (undiluted substrate). Under these conditions, the permeates of the reactor test with 20 RT (diluted substrate) consisted of 80 % POS fractions while that of 40 RT (undiluted substrate) resulted in 70% POS fractions. A two-step continuous biocatalytic MER for the continuous POS production looks very promising for the continuous production of tailor made POS. Although both the processes i.e 20 RT (diluted substrate) and 40 RT (undiluted substrate) gave the best results, but for an Industrial application it is preferable to use an undiluted substrate.

Keywords: pectic oligosaccharides, membrane reactor, residence time, specific productivity, volumetric productivity

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Authors: A. Ojha, Y. K. Gupta

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Organophosphate (OP) pesticides are among the most widely used synthetic chemicals for controlling a wide variety of pests throughout the world. Chlorpyrifos (CPF), methyl parathion (MPT), and malathion (MLT) are among the most extensively used OP pesticides in India. DNA strand breaks and DNA-protein crosslinks (DPC) are toxic lesions associated with the mechanisms of toxicity of genotoxic compounds. In the present study, we have examined the potential of CPF, MPT, and MLT individually and in combination, to cause DNA strand breakage and DPC formation. Peripheral blood lymphocytes of rat were exposed to 1/4 and 1/10 LC50 dose of CPF, MPT, and MLT for 2, 4, 8, and 12h. The DNA strand break was measured by the comet assay and expressed as DNA damage index while DPC estimation was done by fluorescence emission. There was significantly marked increase in DNA damage and DNA-protein crosslink formation in time and dose dependent manner. It was also observed that MPT caused the highest level of DNA damage as compared to other studied OP compounds. Thus, from present study, we can conclude that studied pesticides have genotoxic potential. The pesticides mixture does not potentiate the toxicity of each other. Nonetheless, additional in vivo data are required before a definitive conclusion can be drawn regarding hazard prediction to humans.

Keywords: organophosphate, pesticides, DNA damage, DNA protein crosslink, genotoxic

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Authors: Izzatul Ulfana, Angga Pratomo Cahyadi, Rimayanti, Widjiati

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Freezing oocyte could cause temperature stress. Temperature stress triggers cell damage. Insulin Transferrin Selenium (ITS) and Heat Shock Protein 70 (HSP70) had been used to prevent damage to the oocyte after freezing. ITS and HSP70 could cause the difference protective effect. The aim of this research was to obtain an effective cryoprotectant for freezing local goat oocyte in cumulus cells change. The research began by collecting the ovary from a local slaughterhouse in Indonesia, aspiration follicle, in vitro maturation and the freezing had been used vitrification method. Examination of the morphology cells by native staining method. Data on the calculation morphology oocyte analyzed by Kruskall-Wallis Test. After the Kruskall-Wallis Test which indicated significance, followed by Mann-Whitney Test to compare between treatment groups. As a result, cryoprotectant ITS has the best culumus cells after warming

Keywords: Insulin Transferrin Selenium, Heat Shock Protein 70, cryoprotectant, vitrification

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Authors: Jiraporn Rojtinnakorn

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ICAM-2 (intercellular adhesion molecule 2) or CD102 (Cluster of Differentiation 102) is type I trans-membrane glycoproteins, composing 2-9 immunoglobulin-like C2-type domains. ICAM-2 plays the particular role in immune response and cell surveillance. It is concerned in innate and specific immunity, cell survival signal, apoptosis, and anticancer. EST clone of ICAM-2, from P. gigas blood cell EST libraries, showed high identity to human ICAM-2 (92%) with conserve region of ICAM N-terminal domain and part of Ig superfamily. Gene and protein of ICAM-2 has been founded in mammals. This is the first report of ICAM-2 in fish.

Keywords: ICAM-2, CD102, Pangasianodon gigas, antitumor

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Authors: Ishaq Kashif, Muhammad Younas, Muhammad Riaz, Mubarak Ali

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Poor feeding management during pre-weaning period is one of the factors resulting in compromised growth of Beetal kids fattened for meat purpose. The main reason for this anomaly may be less milk offered to kids and non-serious efforts for its management. This study was planned to find the most appropriate protein level suiting the age of the weaning while shifting animals to high input feeding system. Total of 42 Beetal male kids having 30 (±10), 60 (±10) and 90 (±10) days of age were selected with 16 in each age group. They were designated as G30, G60 and G90, respectively. The weights of animals were; 8±2 kg (G30), 12±2 kg (G60) and 16±2 kg (G90), respectively. All animals were weaned by introducing the total mix feed gradually and withdrawing the milk during the adjustment period of two weeks. The pelleted starter ration (total mix feed) with three various dietary protein levels designated as R1 (16% CP), R2 (20% CP) and R3 (26% CP) were introduced. The control group was reared on the fodder (Maize). The starter rations were iso-caloric and were offered for six-week duration. All animals were exposed to treatment using two-factor factorial (3×3) plus control treatment arrangement under completely randomized design. The data were collected on average daily feed intake (ADFI), average daily gain (ADG), gain to intake ratio, Klieber ratio (KR), body measurements and blood metabolites of kids. The data was analyzed using aov function of R-software. The statistical analysis showed that starter feed protein levels and age of weaning had significant interaction for ADG (P < 0.001), KR (P < 0.001), ADFI (P < 0.05) and blood urea nitrogen (P < 0.05) while serum creatinine and feed conversion had non-significant interaction. The trend analysis revealed that ADG had significant quadratic interaction (P < 0.05) within protein levels and age of weaning. It was found that animals weaned at 30 or 60 days, on R2 diet had better ADG (46.8 gm/day and 87.06 gm/day, respectively) weaned at 60 days of age. The animals weaned at 90 days had best ADG (127 gm/day) with R1. It is concluded that animal weaned at 30 or 40 days required 20% CP for better growth performance while animal at 90 days showed better performance with 16% CP.

Keywords: average daily gain, starter protein levels, weaning age, gain to intake ratio

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Authors: Eitedal Daoud, Reda M.Daoud, Khaled Abdel-Wahhab, Maha M.Saber, Lobna Saber

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Background: Cis-platin is a widely used chemotherapeutic drug to treat many malignant disorders including head and neck malignancies. Oto-nephrotxicity is an important and dose - limiting side effect of cis - platin therapy. Nowadays, more attention had been paid to oto-toxicity caused with cis-platin. Aim of the Work: This study was designed to investigate the potential protective effect of Whey protein (WP) against cis-platin induced ototoxicity compared to the effect of N-acetylcysteine (NAC) in rats. Methodology: Male albino rats were randomly divided into 6 groups: untreated rats (control), rats orally treated with whey protein (1g/kg b.w/day) for seven executive days, rats treated orally with N-acetylcysteine (500 mg/kgb.w /day) for seven executive days, rates intoxicated intraperitoneal (ip) with cis- platin (10 mg/kgb.w. once), rats treated with whey protein (1g/kgb.w./day) for seven executive days) followed by one injection (ip) of cis-platin(10 mg/kg b.w.) one hour after the last oral administration of whey protein, rats treated with N- acetylcysteine (for seven executive days followed by one injection (ip) of cis-platin (10 mg/kgb.w) one hour after the last oral administration of N-acetylcysteine. The organ of Corti, the stria vascularis and spiral ganglia were visualized by light microscopy at different magnifications. Results: Cis-platin intoxicated animals showed a significant decrease in serum level of total antioxidant capacity (TAC),with inhibition in the activity of serum glutathione-s transferase(GST) and paraoxonnase-1 (PON-1) in comparison with control. Group treated with either NAC or WP with cis-platin showed significant elevation in the activity of both GST & PON-1 with increased serum level of TAC when compared with cis-platin intoxicated rats. Animals treated with NAC or WP with cis-platin compared to those treated with cis-platin alone showed marked degree of improvement towards control rats as there was significant drop in the serum level of cortecosterone, nitric oxide (NO), and melandialdehyde (MDA).Histopathologic, in NAC pretreated group there was no changes in stria vascularis or spiral ganglia. In group pretreated with WP, there was no histopathologic alteration detected in the organ of Corti and Reissers membrane but oedema and haemorrhage were founded in the stria vascularis in small focal manner. Conclusion: Our finding showed that Whey protein is a natural dietary supplement product proved its ability of protection of anti-oxidant system and the cochlea against cis-platin induced ototoxicity.

Keywords: anti-oxidant, cis-platin, N-acetylcysteine, ototoxicity, whey protein

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Authors: Chieh-Chung Tsou, Chun-Min Lo, Yeh-Shiu Chu

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Cell’s mechanical forces provide important physical cues in regulation of proper cellular functions, such as cell differentiation, proliferation and migration. It is believed that adhesive forces generated by cell-cell interaction are able to transmit to the interior of cell through filamentous cortical cytoskeleton. Prominent among other membrane receptors, Cadherins are prototypical adhesive molecules able to generate remarkable forces to regulate intercellular adhesion. However, the mechanistic steps of mechano-transduction in Cadherin-mediated adhesion remain very controversial. We are interested in understanding how Cadherin protein complexes enable force generation and transmission at cell-cell contact in the initial stage of intercellular adhesion. For providing a better control of time, space, and substrate stiffness, in this study, a combination of protein micropattern, micropipette manipulation, and traction force microscopy is used. Pair micropattern with different forms confines cell spreading area and the gaps in pairs varied from 2 to 8 microns are applied for monitoring the forces that cell pairs generated, measured by traction force microscopy. Moreover, cell clones obtained from epithelial cells undergone genome editing are used to score the importance for known components of Cadherin complexes in force generation. We believe that our results from this combinatory mechanobiological method will provide deep insights on understanding the biophysical principle governing mechano- transduction of Cadherin-mediated intercellular adhesion.

Keywords: cadherin, intercellular adhesion, protein micropattern, traction force microscopy

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Authors: M. Latha, Achintya K. Dolui, P. Vijayaraj

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Vegetable oils mainly triacylglycerol (TAG) are an essential nutrient in the human diet as well as one of the major global commodity. There is a pressing need to enhance the yield of oil production to meet the world’s growing demand. Oil content is controlled by the balance between synthesis and breakdown in the cells. Several studies have established to increase the oil content by the overexpression of oil biosynthetic enzymes. Interestingly the significant oil accumulation was observed with impaired TAG hydrolysis. Unfortunately, the structural, as well as the biochemical properties of the lipase enzymes, is widely unknown, and so far, no candidate gene was identified in seeds except sugar-dependent1 (SDP1). Evidence has shown that SDP1directly responsible for initiation of oil breakdown in the seeds during germination. The present study is the identification of seed-specific acyl-hydrolases by activity based proteome profiling (ABPP) using Arabidopsis thaliana as a model system. The ABPP reveals that around 8 to 10 proteins having the serine hydrolase domain and are expressed during germination of Arabidopsis seed. The N-term sequencing, as well as LC-MS/MS analysis, was performed for the differentially expressed protein during germination. The coding region of the identified proteins was cloned, and lipases activity was assessed with purified recombinant protein. The enzyme assay was performed against various lipid substrates, and we have observed the acylhydrolase activity towards lysophosphatidylcholine and monoacylglycerol. Further, the functional characteristic of the identified protein will reveal the physiological significance the enzyme in oil accumulation.

Keywords: lipase, lipids, vegetable oil, triacylglycerol

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Authors: Jineetkumar Gawad, Chandrakant Bonde

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Tuberculosis (TB) is a major worldwide concern whose control has been exacerbated by HIV, the rise of multidrug-resistance (MDR-TB) and extensively drug resistance (XDR-TB) strains of Mycobacterium tuberculosis. The interest for newer and faster acting antitubercular drugs are more remarkable than any time. To search potent compounds is need and challenge for researchers. Here, we tried to design lead for inhibition of Decaprenyl phosphoryl-β-D-ribose 2′-epimerase (DprE1) enzyme. Arabinose is an essential constituent of mycobacterial cell wall. DprE1 is a flavoenzyme that converts decaprenylphosphoryl-D-ribose into decaprenylphosphoryl-2-keto-ribose, which is intermediate in biosynthetic pathway of arabinose. Latter, DprE2 converts keto-ribose into decaprenylphosphoryl-D-arabinose. We had a selection of 23 compounds from azaindole series for computational study, and they were drawn using marvisketch. Ligands were prepared using Maestro molecular modeling interface, Schrodinger, v10.5. Common pharmacophore hypotheses were developed by applying dataset thresholds to yield active and inactive set of compounds. There were 326 hypotheses were developed. On the basis of survival score, ADRRR (Survival Score: 5.453) was selected. Selected pharmacophore hypotheses were subjected to virtual screening results into 1000 hits. Hits were prepared and docked with protein 4KW5 (oxydoreductase inhibitor) was downloaded in .pdb format from RCSB Protein Data Bank. Protein was prepared using protein preparation wizard. Protein was preprocessed, the workspace was analyzed using force field OPLS 2005. Glide grid was generated by picking single atom in molecule. Prepared ligands were docked with prepared protein 4KW5 using Glide docking. After docking, on the basis of glide score top-five compounds were selected, (5223, 5812, 0661, 0662, and 2945) and the glide docking score (-8.928, -8.534, -8.412, -8.411, -8.351) respectively. There were interactions of ligand and protein, specifically HIS 132, LYS 418, TRY 230, ASN 385. Pi-pi stacking was observed in few compounds with basic Imidazo [4,5-b] pyridine ring. We had basic azaindole ring in parent compounds, but after glide docking, we received compounds with Imidazo [4,5-b] pyridine as a basic ring. That might be the new lead in the process of drug discovery.

Keywords: DprE1 inhibitors, in silico drug designing, imidazo [4, 5-b] pyridine, lead, tuberculosis

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Authors: M. R. Jokanović, V. M. Tomović, M. T. Jović, S. B. Škaljac, B. V. Šojić, P. M. Ikonić, T. A. Tasić

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Proximate (moisture, protein, total fat, total ash) and mineral (K, P, Na, Mg, Ca, Zn, Fe, Cu and Mn) composition of chicken giblets (heart, liver and gizzard) were investigated. Phosphorous content, as well as proximate composition, were determined according to recommended ISO methods. The content of all elements, except phosphorus, of the giblets tissues were determined using inductively coupled plasma-optical emission spectrometry (ICP-OES), after dry ashing mineralization. Regarding proximate composition heart was the highest in total fat content, and the lowest in protein content. Liver was the highest in protein and total ash content, while gizzard was the highest in moisture and the lowest in total fat content. Regarding mineral composition liver was the highest for K, P, Ca, Mg, Fe, Zn, Cu, and Mn, while heart was the highest for Na content. The contents of almost all investigated minerals in analysed giblets tissues of chickens from Vojvodina were similar to values reported in the literature, i.e. in national food composition databases of other countries.

Keywords: chicken giblets, proximate composition, mineral composition, inductively coupled plasma-optical emission spectrometry (ICP-OES)

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Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko

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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.

Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology

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Authors: Yohannes Kelifa, Gomathi Periasamy, Aman Karim

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Introduction: Diabetes mellitus has become a major public health and economical problem across the globe. Modern antidiabetic drugs have a number of limitations, and scientific investigation of traditional herbal remedies used for diabetes may provide novel leads for the development of new antidiabetic drugs that can be used as alternative or complementary to available antidiabetic allopathic medications. Though Myrica salicifolia Hochst. ex A. Rich. is used for the management of diabetes in Ethiopian traditional medicine, there was no previous scientific evidence about its antidiabetic effect to the authors’ knowledge. This study was undertaken to evaluate the antidiabetic activity the root extracts of Myrica salicifolia in streptozotocin (STZ)-induced diabetic mice. Methods: Experimental diabetes was induced by intraperitoneal administration of STZ (150 mg/kg) in male mice. Diabetic mice were treated with oral doses of M. salicifolia root extracts at 200, 400 and 600 mg/kg, and its fractions (chloroform, ethyl acetate, n-butanol and aqueous) at a dose of 400 mg/kg daily for 15 days. Fasting blood glucose level (BGL) was measured at 0, 5th,10th, and 15th day. The free radical scavenging activity of the crude extract was determined using in vitro by DPPH assay. The statistical significance was assessed by one-way ANOVA, followed by Tukey’s multiple comparison tests. Results were considered significant when p < 0.05. Results: Daily administration of the M. salicifolia 80% methanol root extracts (at three different doses (200, 400 and 600 mg/kg) significantly (p < 0.05, p < 0.01 and p < 0.001) reduced fasting BGL compared with diabetic control. The aqueous and butanol fractions at a dose of 400 mg/kg resulted in maximum reduction of fasting BGL by 42.39%, and 52.13%, respectively at the 15th day in STZ-induced diabetic mice. Free radical scavenging activity of the 80% methanol extract of M. salicifolia was comparable to ascorbic acid. The IC50 values of the crude extract and ascorbic acid (a reference compound) were found to be 4.54 μg/ml and 4.39 μg/ml, respectively. Conclusion: These findings demonstrated that the methanolic extracts of M. salicifolia root and its fractions (n-butanol and aqueous) exhibit a significant antihyperglycemic activity in STZ-induced diabetic mice. Furthermore, the result of the present study indicates that M. salicifolia root extract is a potential source of natural antioxidants.

Keywords: antidiabetic, diabetes mellitus, DPPH, mice, Myrica salicifolia, streptozotocin

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Authors: Jesus Hernandez, Zead Elzoeiry, Md. S. Islam, Abel E. Navarro

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The adsorption of bovine serum albumin (BSA) and human hemoglobin (Hb) on naturally-occurring adsorbents was studied to evaluate the potential recovery of proteins from meat industry residues. Spent peppermint tea (PM), powdered purple corn cob (PC), natural clay (NC) and chemically-modified clay (MC) were investigated to elucidate the effects of pH, adsorbent dose, initial protein concentration, presence of salts and heavy metals. Equilibrium data were fitted according to isotherm models, reporting a maximum adsorption capacity at pH 8 of 318 and 344 mg BSA/g of PM and NC, respectively. Moreover, Hb displayed maximum adsorption capacity at pH 5 of 125 and 143 mg/g of PM and PC, respectively. Hofmeister salt effect was only observed for PM/Hb system. Salts tend to decrease protein adsorption, and the presence of Cu(II) ions had negligible impacts on the adsorption onto NC and PC. Desorption experiments confirmed that more than 85% of both proteins can be recovered with diluted acids and bases. SEM, EDX, and TGA analyses demonstrated that the adsorbents have favorable morphological and mechanical properties. The long-term goal of this study aims to recover soluble proteins from industrial wastewaters to produce animal food or any protein-based product.

Keywords: adsorption, albumin, clay, hemoglobin, spent peppermint leaf

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Authors: Sanchir Erdenebayar, Namuuntsetseg Oyunbaatar

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Objectives: Peritoneal dialysis treatment is one of the important forms of kidney replacement therapy, and it has recently developed instantly in Mongolia for the past five years. Currently, more than 120 patients undergo peritoneal dialysis nationwide. These patients lack nutritional education, which predisposes them to protein deficiency and further impairs their quality of life. However, there is no study which is conducted among those about their dietary in Mongolia. Therefore, integrated nutrition information and educating them about dietary patterns to follow are urgently needed for PD patients. Methods: A cross-sectional study was carried out on 45 patients aged between 18 and 60 years who were undergoing CAPD at the biggest Medvic dialysis center in Ulaanbaatar. The knowledge of nutrition and food intake is assessed by interview based on a validated questionnaire prepared from KDIGO guidelines, semi-FFQ and a 24-hour dietary recall method. In addition, a biochemical blood test that includes total protein, albumin, calcium, phosphorus, potassium, and hemoglobin is used for an assessment of the patient’s current nutritional status. Results: Knowledge of nutritional status for CAPD was great, with 21.4% of patients and 78.65% having poor nutrition knowledge. The rate of mild to moderate malnutrition was 48.8% among research participants. Serum albumin was 38.4 ± 4.7 g/L, and total protein was 67.3±7.5g/l. Patients met 62.5± 26.5% of their daily intake nutritional requirement for calories and 72±40% of their nutritional requirement for protein. All patients’ energy intake was significantly /1328±304kcal/ lower than the energy requirement (2124±378kcal). Only 14.2% met the recommended dietary protein intake recommended to them of greater than 1.2 g/kg. Conclusions: As was established before, nutritional education has a vital positive impact on the health and nutritional status of peritoneal dialysis patients. The results of this study show that nutritional education programs are not enough adequate in peritoneal dialysis patients. There is a crucial priority to establish nutritional educational programs and guidelines for PD patients in Mongolia.

Keywords: renal diet, peritoneal dialysis, nutrition education, CKD diet

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Authors: Nuri Başpınar, Abdullah Başoğlu, Özgür Özdemir, Çağlayan Özel, FundaTerzi, Özgür Yaman

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Current research is targeting new molecular mechanisms that underlie non-alcoholic fatty liver disease (NAFLD) and associated metabolic disorders like nonalcoholic steatohepatitis (NASH). Forty New Zealand White rabbits have been used and fed a high protein (HP) and energy diet based on grains and containing 11.76 MJ/kg. Boron added to 3 experimental groups’ drinking waters (30 mg boron/L) as boron compounds. Biochemical analysis including boron levels, and nuclear magnetic resonance (NMR) based metabolomics evaluation, and mRNA expression of peroxisome proliferator-activated receptor (PPAR) family were performed. LDL-cholesterol concentrations alone were decreased in all the experimental groups. Boron levels in serum and feces were increased. Content of acetate was in about 2x higher for anhydrous borax group, at least 3x higher for boric acid group. PPARα mRNA expression was significantly decreased in boric acid group. Anhydrous borax attenuated mRNA levels of PPARα, which was further suppressed by boric acid. Boron supplementation decreased the degenerative alterations in hepatocytes. Except borax group other boron groups did not have a pronounced change in tubular epithels of kidney. In conclusion, high protein and energy diet leads hepatocytes’ degenerative changes which can be prevented by boron supplementation. Boric acid seems to precede in this effectiveness.

Keywords: high protein and energy diet, boron, metabolomics, transcriptomic

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Authors: O. D. A. N. Perera, H. G. Wanigasinghe

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Tender jackfruit is an underutilized biomass, which still has a good consumer demand. Valorization of this ingredient into meat analog would obtain greater consumer acceptance due to concerns about health, the environment, and living sustainably of mankind have increased significantly in this decade, opening the market for meat substitutes. The objective of this research was to create a plant-based meat substitute with a structure similar to meat products. In this study, three different combinations of tender jackfruit were used to create vegan burger patties, which were then examined for their textural, physico-chemical, and sensory qualities. The developed burger patties have been compared with store-bought chicken patties. The developed vegan burger patties P1, P2, and P3 had a comparable flavor preference to the control and demonstrated considerable general acceptability (p >.05). P3 has a high quantity of protein (17.10 ± 0.02%) and fiber (6.40 ± 0.06%). At the same time, the vegan burger patty resulted in less fat, high fiber, and high protein which meets the vegan consumer requirements.

Keywords: underutilized, high fibre, soya protein isolate, cooking yield

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Authors: Othman E. Othman, Amira M. Nowier, Medhat El-Denary

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Camels play an important socio-economic role within the pastoral and agricultural system in the dry and semidry zones of Asia and Africa. Camels are economically important animals in Egypt where they are dual purpose animals (meat and milk). The analysis of chemical composition of camel milk showed that the total protein contents ranged from 2.4% to 5.3% and it is divided into casein and whey proteins. The casein fraction constitutes 52% to 89% of total camel milk protein and it divided into 4 fractions namely αs1, αs2, β and κ-caseins which are encoded by four tightly genes. In spite of the important role of casein genes and the effects of their genetic polymorphisms on quantitative traits and technological properties of milk, the studies for the detection of genetic polymorphism of camel milk genes are still limited. Due to this fact, this work focused - using PCR-RFP and sequencing analysis - on the identification of genetic polymorphisms and SNPs of two casein genes in Maghrabi camel breed which is a dual purpose camel breed in Egypt. The amplified fragments at 488-bp of the camel κ-CN gene were digested with AluI endonuclease. The results showed the appearance of three different genotypes in the tested animals; CC with three digested fragments at 203-, 127- and 120-bp, TT with three digested fragments at 203-, 158- and 127-bp and CT with four digested fragments at 203-, 158-, 127- and 120-bp. The frequencies of three detected genotypes were 11.0% for CC, 48.0% for TT and 41.0% for CT genotypes. The sequencing analysis of the two different alleles declared the presence of a single nucleotide polymorphism (C→T) at position 121 in the amplified fragments which is responsible for the destruction of a restriction site (AG/CT) in allele T and resulted in the presence of two different alleles C and T in tested animals. The nucleotide sequences of κ-CN alleles C and T were submitted to GenBank with the accession numbers; KU055605 and KU055606, respectively. The primers used in this study amplified 942-bp fragments spanning from exon 4 to exon 6 of camel αS1-Casein gene. The amplified fragments were digested with two different restriction enzymes; SmlI and AluI. The results of SmlI digestion did not show any restriction site whereas the digestion with AluI endonuclease revealed the presence of two restriction sites AG^CT at positions 68^69 and 631^632 yielding the presence of three digested fragments with sizes 68-, 563- and 293-bp.The nucleotide sequences of this fragment from camel αS1-Casein gene were submitted to GenBank with the accession number KU145820. In conclusion, the genetic characterization of quantitative traits genes which are associated with the production traits like milk yield and composition is considered an important step towards the genetic improvement of livestock species through the selection of superior animals depending on the favorable alleles and genotypes; marker assisted selection (MAS).

Keywords: genetic polymorphism, SNP polymorphism, Maghrabi camels, κ-Casein gene, αS1-Casein gene

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Authors: Shikha Masand, Sudesh Kumar Yadav

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Certain protein kinases have been shown to be crucial for plant cell signaling pathways associated with plant immune responses. Here we identified a horsegram [Macrotyloma uniflorum (Lam.) Verdc.] malectin-like leucine rich receptor-like protein kinase (RLK) gene MuRLK. The functional MuRLK protein preferentially binds to mannose and N-acetyl glucosamine residues. MuRLK exists in the cytoplasm and also localizes to the plasma membrane of plant cells via its N-terminus. Over-expression of MuRLK in Arabidopsis enhances the basal resistance to infection with Pseudomonas syringae pv. tomato, Alternaria brassicicola and Hyaloperonospora arabidopsidis, are associated with elevated ROS bursts, MAPK activation, thus ultimately leading to hypersensitive cell death. Moreover, salicylic acid-dependent and jasmonic acid-dependent defense responses are also enhanced in the MuRLK-overexpressed plants that lead to HR-induced cell death. Together, these results suggest that MuRLK plays a key role in the regulation of plant cell death, early and late defense responses after the recognition of microbial pathogens.

Keywords: horsegram, Pseudomonas syringae pv. tomato, MuRLK, ROS burst, cell death, plant defense

Procedia PDF Downloads 248

Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

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Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity

Procedia PDF Downloads 190

Authors: Farramae Francisco, Rhoda Mae Simora, Sharon Nunal

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Deproteination and demineralization efficiencies of shrimp waste using two Lactobacillus species treated with different carbohydrate sources for chitin production, its chemical conversion to chitosan and the quality of chitin and chitosan produced were determined. Using 5% glucose and 5% cassava starch as carbohydrate sources, pH slightly increased from the initial pH of 6.0 to 6.8 and 7.2, respectively after 24 h and maintained their pH at 6.7 to 7.3 throughout the treatment period. Demineralization (%) in 5 % glucose and 5 % cassava was highest during the first day of treatment which was 82% and 83%, respectively. Deproteination (%) was highest in 5% cassava starch on the 3rd day of treatment at 84.4%. The obtained chitin from 5% cassava and 5% glucose had a residual ash and protein below 1% and solubility of 59% and 44.3%, respectively. Chitosan produced from 5% cassava and 5% glucose had protein content below 0.05%; residual ash was 1.1% and 0.8%, respectively. Chitosan solubility and degree of deacetylation were 56% and 33% in 5% glucose and 48% and 29% in 5% cassava, respectively. The advantage this alternative technology offers over that of chemical extraction is large reduction in chemicals needed thus less effluent production and generation of a protein-rich liquor, although the demineralization process should be improved to achieve greater degree of deacetylation.

Keywords: alternative carbon source, bioprocessing, lactic acid bacteria, waste utilization

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Authors: Lineda Rouissat-Dahane, Abdelkrim Cheriti, Abbderazak Marouf, Reddy Kandappa H., Govender Patrick

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Natural phytochemicals extracted from folk herbal have drawn much attention in complementary and alternative medicine, and the plant kingdom is considered for developing new molluscicide. The aqueous and acetone extract of the aerial parts of some Asteraceae (Anvillea radiata, Bubonium graveolens, Launaea arborescens, Launaea nudicaulis and Warionia saharae) were investigated for its molluscicidal activity against Lymnaea acuminata showed significant molluscicidal activity with a median lethal concentration (LC50) of aqueous extract (8,178mg/ml) and organic extract 0.002μg/mL, which was indicated higher potency than the positive control, (LC50=100mg /mL for aqueous extract ; LC50=11.6 μg/mL for organic extract). Among the extract and their fractions, those of aerial parts of Launaea nudicaulis and Warionia saharae were found to exhibit significant molluscicidal activities. Among different solvent fractions of the acetone extract of Warionia saharae, the dichloromethane (DCM) soluble fraction showed the most potent molluscicidal activity against Lymnaea acuminata. Plants in species Anvillea radiata, Bubonium graveolens, Launaea arborescens, Launaea nudicaulis, and Warionia saharae produce a great variety of Flavonoids, Glucoside flavonoids, and Saponins that confer natural resistance against several pests. Most extracts were found to exhibit significant molluscicidal activity.

Keywords: acetone extract, aqueous extract, Asteraceae, molluscicidal activity, Lymnaea acuminata

Procedia PDF Downloads 131