Search results for: transmembrane protein

Authors: Pramod Somvanshi, Manu Tomar, K. V. Venkatesh

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Insulin and glucagon are responsible for homeostasis of key plasma metabolites like glucose, amino acids and fatty acids in the blood plasma. These hormones act antagonistically to each other during the secretion and signaling stages. In the present work, we analyze the effect of macronutrients on the response from integrated insulin and glucagon signaling pathways. The insulin and glucagon pathways are connected by DAG (a calcium signaling component which is part of the glucagon signaling module) which activates PKC and inhibits IRS (insulin signaling component) constituting a crosstalk. AKT (insulin signaling component) inhibits cAMP (glucagon signaling component) through PDE3 forming the other crosstalk between the two signaling pathways. Physiological level of anabolism and catabolism is captured through a metric quantified by the activity levels of AKT and PKA in their phosphorylated states, which represent the insulin and glucagon signaling endpoints, respectively. Under resting and starving conditions, the phosphorylation metric represents homeostasis indicating a balance between the anabolic and catabolic activities in the tissues. The steady state analysis of the integrated network demonstrates the presence of a bistable response in the phosphorylation metric with respect to input plasma glucose levels. This indicates that two steady state conditions (one in the homeostatic zone and other in the anabolic zone) are possible for a given glucose concentration depending on the ON or OFF path. When glucose levels rise above normal, during post-meal conditions, the bistability is observed in the anabolic space denoting the dominance of the glycogenesis in liver. For glucose concentrations lower than the physiological levels, while exercising, metabolic response lies in the catabolic space denoting the prevalence of glycogenolysis in liver. The non-linear positive feedback of AKT on IRS in insulin signaling module of the network is the main cause of the bistable response. The span of bistability in the phosphorylation metric increases as plasma fatty acid and amino acid levels rise and eventually the response turns monostable and catabolic representing diabetic conditions. In the case of high fat or protein diet, fatty acids and amino acids have an inhibitory effect on the insulin signaling pathway by increasing the serine phosphorylation of IRS protein via the activation of PKC and S6K, respectively. Similar analysis was also performed with respect to input amino acid and fatty acid levels. This emergent property of bistability in the integrated network helps us understand why it becomes extremely difficult to treat obesity and diabetes when blood glucose level rises beyond a certain value.

Keywords: bistability, diabetes, feedback and crosstalk, obesity

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Authors: S. Paengkoum, P. Paengkoum, W. Kaewwongsa

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Twenty-four male growing goats were randomly assigned to a Randomized Complete Block Design. Dietary treatments were different level of feeding concentrate diet at 1.0, 1.5, 2.0, and 2.5% of body weight (BW). The results showed that average daily gain, microbial N supply, N retention of meat goats in the group of feeding level at 2.0% BW and 2.5% BW were significantly higher (P<0.05) than those goats fed with feeding levels of 1.0% BW and 1.5% BW. Based on this result the conclusion can be made that using 75% fermented cassava pulp by Saccharomyces cerevisiae as the main source of protein to completely replace soybean meal was beneficial to meat goats in terms of feed intake. The feeding concentrate at levels between 2.0-2.5% BW gives highest in the growth of meat goat in this experiment.

Keywords: cassava pulp, yeast, goat, nitrogen retention

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Authors: Oyewusi Peter Ayodele, Onipede Ayodeji

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Our research focuses on some physicochemical parameters of Moringa Oleifera seed meal and its seed oil to determine its nutritional quality. Proximate, mineral, and vitamin analyses were performed on the defatted seed meal, while fatty acid determination was carried out on the seed oil. The results of the proximate composition show moisture content (3.52 ± 0.01), ash (2.80 ± 0.33), crude fibre (3.92 ± 0.01), protein (42.96 ± 0.05), crude fat (7.04 ± 0.01) and carbohydrate (36.79 ± 0.04). The mineral composition shows that the seed is rich in Ca, K, and Na with 220ppm, 205ppm, and 118ppm, respectively. The seed has vitamins A and C with 2.17 ± 0.01mg/100g and 6.95 ± 0.00 mg/100g respectively. The seed also contains 56.62 %, 38.50 %, and 5.24 % saturated, monounsaturated, and polyunsaturated fatty acids, respectively. It could be illustrated that Moringa seeds and their oil can be considered potential sources for both dietary and industrial purposes.

Keywords: Moringa oleifera seed, chemical composition, fatty acid, proximate, minerals and vitamins compositions

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Authors: Mesfin Angaw Tesfay

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Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes, serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein, two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli, and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg, respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, glycosylation

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Authors: Aastha Arora, Vikas Bhuria, Saurabh Singh, Uma Pathak, Shweta Mathur, Puja P. Hazari, Rajat Sandhir, Ravi Soni, Anant N. Bhatt, Bilikere S. Dwarakanath

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Radiotherapy is an effective curative and palliative option for patients with thoracic malignancies. However, lung injury, comprising of pneumonitis and fibrosis, remains a significant clin¬ical complication of thoracic radiation, thus making it a dose-limiting factor. Also, injury to the lung is often reported as part of multi-organ failure in victims of accidental radiation exposures. Radiation induced inflammatory response in the lung, characterized by leukocyte infiltration and vascular changes, is an important contributing factor for the injury. Therefore, countermeasure agents to attenuate radiation induced inflammatory response are considered as an important approach to prevent chronic lung damage. Although Amifostine, the widely used, FDA approved radio-protector, has been found to reduce the radiation induced pneumonitis during radiation therapy of non-small cell lung carcinoma, its application during mass and field exposure is limited due to associated toxicity and ineffectiveness with the oral administration. The amifostine analogue (DRDE-30) overcomes this limitation as it is orally effective in reducing the mortality of whole body irradiated mice. The current study was undertaken to investigate the potential of DRDE-30 to ameliorate radiation induced lung damage. DRDE-30 was administered intra-peritoneally, 30 minutes prior to 13.5 Gy thoracic (60Co-gamma) radiation in C57BL/6 mice. Broncheo- alveolar lavage fluid (BALF) and lung tissues were harvested at 12 and 24 weeks post irradiation for studying inflammatory and fibrotic markers. Lactate dehydrogenase (LDH) leakage, leukocyte count and protein content in BALF were used as parameters to evaluate lung vascular permeability. Inflammatory cell signaling (p38 phosphorylation) and anti-oxidant status (MnSOD and Catalase level) was assessed by Western blot, while X-ray CT scan, H & E staining and trichrome staining were done to study the lung architecture and collagen deposition. Irradiation of the lung increased the total protein content, LDH leakage and total leukocyte count in the BALF, reflecting endothelial barrier dysfunction. These disruptive effects were significantly abolished by DRDE-30, which appear to be linked to the DRDE-30 mediated abrogation of activation of the redox-sensitive pro- inflammatory signaling cascade, the MAPK pathway. Concurrent administration of DRDE-30 with radiation inhibited radiation-induced oxidative stress by strengthening the anti-oxidant defense system and abrogated p38 mitogen-activated protein kinase activation, which was associated with reduced vascular leak and macrophage recruitment to the lungs. Histopathological examination (by H & E staining) of the lung showed radiation-induced inflammation of the lungs, characterized by cellular infiltration, interstitial oedema, alveolar wall thickening, perivascular fibrosis and obstruction of alveolar spaces, which were all reduced by pre-administration of DRDE-30. Structural analysis with X-ray CT indicated lung architecture (linked to the degree of opacity) comparable to un-irradiated mice that correlated well with the lung morphology and reduced collagen deposition. Reduction in the radiation-induced inflammation and fibrosis brought about by DRDE-30 resulted in a profound increase in animal survival (72 % in the combination vs 24% with radiation) observed at the end of 24 weeks following irradiation. These findings establish the potential of the Amifostine analogue, DRDE-30, in reducing radiation induced pulmonary injury by attenuating the inflammatory and fibrotic responses.

Keywords: amifostine, fibrosis, inflammation, lung injury radiation

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Authors: Mounir Soule, Hindatou Saidou, Razafimahefa, Mohamed Thani Ibouroi

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High levels of habitat fragmentation and loss are the main drivers of plant species extinction. They reduce the habitat quality, which is a determining factor for the reproduction of plant species, and generate strong selective pressures for habitat selection, with impacts on the reproduction and survival of individuals. The Comorian’s yam (Dioscorea comorensis) is one of the most threatened plant species of the Comoros archipelago. The species faces one of the highest rates of habitat loss worldwide (9.3 % per year) and is classified as Endangered in the IUCN red list. Despite the nutritional potential of this tuber, the Comorian’s yam cultivation remains neglected by local populations due probably to lack of knowledge on its nutritional importance and the factors driving its spatial distribution and development. In this study, we assessed the nutritional characteristics of Dioscorea comorensis and the drivers of spatial distribution and abundance to propose conservation measures and improve crop yields. To determine the nutritional characteristics, the Kjeldahl method, the Soxhlet method, and Atwater's specific calorific coefficients methods were applied for analyzing proteins, lipids, and caloric energy respectively. In addition, atomic absorption spectrometry was used to measure mineral particles. By combining species occurrences with ecological (habitat types), climatic (temperature, rainfall, etc.), and physicochemical (soil types and quality) variables, we assessed habitat suitability and spatial distribution of the species and the factors explaining the origin, persistence, distribution and competitive capacity of a species using a Species Distribution Modeling (SDM) method. The results showed that the species contains 83.37% carbohydrates, 6.37% protein, and 0.45% lipids. In 100 grams, the quantities of Calcium, Sodium, Zinc, Iron, Copper, Potassium, Phosphorus, Magnesium, and Manganese are respectively 422.70, 599.41, 223.11, 252.32, 332.20, 780.41, 444.17, 287.71 and 220.73 mg. Its PRAL index is negative (- 9.80 mEq/100 g), and its Ca/P (0.95) and Na/K (0.77) ratios are less than 1. This species provides an energy value of 357.46 Kcal per 100 g, thanks to its carbohydrates and minerals and is distinguished from others by its high protein content, offering benefits for cardiovascular health. According to our SDM, the species has a very limited distribution, restricted to forests with higher biomass, humidity, and clay. Our findings highlight how distribution patterns are related to ecological and environmental factors. They also emphasize how the Comoros yam is beneficial in terms of nutritional quality. Our results represent a basic knowledge that will help scientists and decision-makers to develop conservation strategies and to improve crop yields.

Keywords: Dioscorea comorensis, nutritional characteristics, species distribution modeling, conservation strategies, crop yields improvement

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Authors: Sandeep Kaur

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Phycoerythrin (PE) from the mesophilic filamentous cyanobacterium Nostoc piscinale PUPCCC 405.17, a good producer of phycobiliproteins, has been characterized in terms of their unit assembly and stability. The phycoerythrin was extracted by freeze-thawing the cells in water, concentrated by ammonium sulphate fractionation and purified by anion exchange chromatography. The purification process resulted in 2.90 fold increase in phycoerythrin purity reaching to 1.54. Sodium Dodecyl Sulphate- Polyacrylamide Gel Electrophoresis of purified PE demonstrated three protein bands of 14.3, 27.54 and 39.81 kDa. The native PE also showed one band of 125.87 kDa, assumed to be a dimer (αβ)2γ based on results of non-denaturing PAGE. Lyophilized powder PE was more stable compared to phycoerythrin in the solution. The half-life of dry PE is 80 days when stored at 4 °C under dark. The phycoerythrin from this organism has potential applications in food as natural colour and as a fluorescent marker.

Keywords: characterization, Nostoc piscinale, phycoerythrin, purification

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Authors: Ranjita Ghoshmoulick, Aswathi Madhavan, Subhavna Juneja, Prasenjit Sen, Jaydeep Bhattacharya

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Type 2 diabetes mellitus (T2DM) is a very complex and multifactorial metabolic disease where the blood sugar level goes up. One of the major consequence of this elevated blood sugar is the formation of AGE (Advance Glycation Endproducts), from a series of chemical or biochemical reactions. AGE are detrimental because it leads to severe pathogenic complications. They are a group of structurally diverse chemical compounds formed from nonenzymatic reactions between the free amino groups (-NH2) of proteins and carbonyl groups (>C=O) of reducing sugars. The reaction is known as Maillard Reaction. It starts with the formation of reversible schiff’s base linkage which after sometime rearranges itself to form Amadori Product along with dicarbonyl compounds. Amadori products are very unstable hence rearrangement goes on until stable products are formed. During the course of the reaction a lot of chemically unknown intermediates and reactive byproducts are formed that can be termed as Early Glycation Products. And when the reaction completes, structurally stable chemical compounds are formed which is termed as Advanced Glycation Endproducts. Though all glycation products have not been characterized well, some fluorescence compounds e.g pentosidine, Malondialdehyde (MDA) or carboxymethyllysine (CML) etc as AGE and α-dicarbonyls or oxoaldehydes such as 3-deoxyglucosone (3-DG) etc as the intermediates have been identified. In this work Gold NanoParticle (GNP) was used as an optical indicator of glycation products. To achieve faster glycation kinetics and high AGE accumulation, fructose was used instead of glucose. Hemoglobin A0 (HbA0) was fructosylated by in-vitro method. AGE formation was measured fluorimetrically by recording emission at 450nm upon excitation at 350nm. Thereafter this fructosylated HbA0 was fractionated by column chromatography. Fractionation separated the proteinaceous substance from the AGEs. Presence of protein part in the fractions was confirmed by measuring the intrinsic protein fluorescence and Bradford reaction. GNPs were synthesized using the templates of chromatographically separated fractions of fructosylated HbA0. Each fractions gave rise to GNPs of varying color, indicating the presence of distinct set of glycation products differing structurally and chemically. Clear solution appeared due to settling down of particles in some vials. The reactive groups of the intermediates kept the GNP formation mechanism on and did not lead to a stable particle formation till Day 10. Whereas SPR of GNP showed monotonous colour for the fractions collected in case of non fructosylated HbA0. Our findings accentuate the use of GNPs as a simple colorimetric sensing platform for the identification of intermediates of glycation reaction which could be implicated in the prognosis of the associated health risk due to T2DM and others.

Keywords: advance glycation endproducts, glycation, gold nano particle, sensor

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Authors: Mbaeyi-Nwaoha, Ifeoma Elizabeth, Orngu, Africa Orngu

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Composite flours were processed from blends of yellow maize (Zea mays), sesame seed (Sesamum indicum) and oyster mushroom (Pleurotus ostreatus) powder in the ratio of 80:20:0; 75:20:5; 70:20:10; 65:20:10 and 60:20:20, respectively to produce the breakfast cereal coded as YSB, SMB, TMB, PMB and OMB with YSB as the control. The breakfast cereals were produced by hydration and toasting of yellow maize and sesame to 160oC for 25 minutes and blended together with oven dried and packaged oyster mushroom. The developed products (flours and breakfast cereals) were analyzed for proximate composition, vitamins, minerals, anti-nutrients, phytochemicals, functional, microbial and sensory properties. Results for the flours showed: proximate composition (%): moisture (2.59-7.27), ash (1.29-7.57), crude fat (0.98-14.91), fibre (1.03-16.02), protein (10.13-35.29), carbohydrate (75.48-38.18) and energy (295.18-410.75kcal). Vitamins ranged as: vitamin A (0.14-9.03 ug/100g), vitamin B1 (0.14-0.38), vitamin B2 (0.07-0.15), vitamin B3(0.89-4.88) and Vitamin C (0.03-4.24). Minerals (mg/100g) were reported thus: calcium (8.01-372.02), potassium (1.40-1.85), magnesium (12.09-13.15), iron (1.23-5.25) and zinc (0.85-2.20). The results for anti-nutrients and phytochemical ranged from: tannin (1.50-1.61mg/g), Phytate (0.40-0.71mg/g), Oxalate(1.81-2.02mg/g), Flavonoid (0.21-1.27%) and phenolic (1.12-2.01%). Functional properties showed: bulk density (0.51-0.77g/ml), water absorption capacity (266.0-301.5%), swelling capacity (136.0-354.0%), least Gelation (0.55-1.45g/g) and reconstitution index (35.20-69.60%). The total viable count ranged from 6.4× 102to1.0× 103cfu/g while the total mold count was from 1.0× 10to 3.0× 10 cfu/g. For the breakfast cereals, proximate composition (%) ranged thus: moisture (4.07-7.08), ash (3.09-2.28), crude fat(16.04-12.83), crude fibre(4.30-8.22), protein(16.14-22.54), carbohydrate(56.34-47.04) and energy (434.34-393.83Kcal).Vitamin A (7.99-5.98 ug/100g), vitamin B1(0.08-0.42mg/100g), vitamin B2(0.06-0.15 mg/100g), vitamin B3(1.91-4.52 mg/100g) and Vitamin C(3.55-3.32 mg/100g) were reported while Minerals (mg/100g) were: calcium (75.31-58.02), potassium (0.65-4.01), magnesium(12.25-12.62), iron (1.21-4.15) and zinc (0.40-1.32). The anti-nutrients and phytochemical revealed the range (mg/g) as: tannin (1.12-1.21), phytate (0.69-0.53), oxalate (1.21-0.43), flavonoid (0.23-1.22%) and phenolic (0.23-1.23%). The bulk density (0.77-0.63g/ml), water absorption capacity (156.5-126.0%), swelling capacity (309.5-249.5%), least gelation (1.10-0.75g/g) and reconstitution index (49.95-39.95%) were recorded. From the total viable count, it ranged from 3.3× 102to4.2× 102cfu/g but no mold growth was detected. Sensory scores revealed that the breakfast cereals were acceptable to the panelist with oyster mushroom supplementation up to 10%.

Keywords: oyster mushroom (Pleurotus ostreatus), sesame seed (Sesamum indicum), yellow maize (Zea mays, instant breakfast cereals

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Authors: Sibel Silici

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Palynological spectrum, proximate and fatty acids composition of eight beebread samples obtained from different geographical origins were determined. Beebread moisture contents varied between 11.4-15.9 %, ash 1.9-2.54 %, fat 5.9-11.5 %, and protein between 14.8-24.3 %. To our knowledge, this is the first study investigating fatty acids (FAs) composition of the selected monofloral beebreads. A total of thirty-seven FAs were identified. Of these (9Z, 12Z, 15Z)-octadeca-9, 12, 15-trienoic acid, (9Z, 12Z)-octadeca-9, 12-dienoic acid, hexadecanoic acid, (Z)-octadec-9-enoic acid, (Z)-icos-11-enoic acid and octadecanoic acid were the most abundant in all the samples. Cotton beebread contained the highest level of ω-3 FAs, 41.3 %. Unsaturated/saturated FAs ratios ranged between 1.38 and 2.39 indicating that beebread is a good source of unsaturated FAs. The pollen, proximate and FAs composition of beebread samples of different botanical and geographical origins varied significantly.

Keywords: bee bread, fatty acid composition, proximate composition, pollen analysis

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Authors: Ruswanto Ruswanto, Richa Mardianingrum, Tita Nofianti, Nur Rahayuningsih

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Tuberculosis (TB) is a chronic disease as a result of Mycobacterium tuberculosis. It can affect all age groups, and hence, is a global health problem that causes the death of millions of people every year. One of the drugs used in tuberculosis treatment is isonicotinohydrazide. In this study, N'-benzoylisonicotinohydrazide derivative compounds (a-l) were prepared using acylation reactions between isonicotinohydrazide and benzoyl chloride derivatives, through the reflux method. Molecular docking studies suggested that all of the compounds had better interaction with Mycobacterium tuberculosis enoyl-acyl carrier protein reductase (InhA) than isonicotinohydrazide. It can be concluded that N'-benzoylisonicotinohydrazide derivatives (a-l) could be used as anti-tuberculosis candidates. From the docking results revealed that all of the compounds interact well with InhA, with compound g (N'-(3-nitrobenzoyl)isonicotinohydrazide) exhibiting the best interaction.

Keywords: anti-tuberculosis , docking, InhA, N'-benzoylisonicotinohydrazide, synthesis

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Authors: Ajuna B. Henry

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In recent decades, climate change has demanded biological pesticides; more Bt strains are being discovered worldwide, some containing novel insecticidal genes while others have been modified through molecular approaches for increased yield, toxicity, and wider host target. In this study, B. thuringiensis strain AH-2 (Bt-2) was isolated from the soil and tested for insecticidal activity against Aphis gossypii (Hemiptera: Aphididae) and Lepidoptera insect pests: fall webworm (Hyphantria cunea) and diamondback moth (Plutella xylostella). A commercial strain B. thuringiensis subsp. kurstaki (Btk), and a chemical pesticide, imidacloprid (for Hemiptera) and chlorantraniliprole (for Lepidoptera), were used as positive control and the same media (without bacterial inoculum) as a negative control. For aphidicidal activity, Bt-2 caused a mortality rate of 70.2%, 78.1% or 88.4% in third instar nymphs of A. gossypii (3N) at 10%, 25% or 50% culture concentrations, respectively. Moreover, Bt-2 was effectively produced in cost-effective (PB) supplemented with either glucose (PBG) or sucrose (PBS) and maintained high aphicidal efficacy with 3N mortality rates of 85.9%, 82.9% or 82.2% in TSB, PBG or PBS media, respectively at 50% culture concentration. Bt-2 also suppressed adult fecundity by 98.3% compared to only 65.8% suppression by Btk at similar concentrations but was slightly lower than chemical treatment, which caused 100% suppression. Partial purification of 60 – 80% (NH4)2SO4 fraction of Bt-2 aphicidal proteins purified on anion exchange (DEAE-FF) column revealed a 105 kDa aphicidal protein with LC50 = 55.0 ng/µℓ. For Lepidoptera pests, chemical pesticide, Bt-2, and Btk cultures, mortality of 86.7%, 60%, and 60% in 3rd instar larvae of P. xylostella, and 96.7%, 80.0%, and 93.3% in 6th instar larvae of H. cunea, after 72h of exposure. When the entomopathogenic strains were cultured in a cost-effective PBG or PBS, the insecticidal activity in all strains was not significantly different compared to the use of a commercial medium (TSB). Bt-2 caused a mortality rate of 60.0%, 63.3%, and 50.0% against P. xylostella larvae and 76.7%, 83.3%, and 73.3% against H. cunea when grown in TSB, PBG, and PBS media, respectively. Bt-2 (grown in cost-effective PBG medium) caused a dose-dependent toxicity of 26.7%, 40.0%, and 63.3% against P. xylostella and 46.7%, 53.3%, and 76.7% against H. cunea at 10%, 25% and 50% culture concentration, respectively. The partially purified Bt-2 insecticidal proteins fractions F1, F2, F3, and F4 (extracted at different ratios of organic solvent) caused low toxicity (50.0%, 40.0%, 36.7%, and 30.0%) against P. xylostella and relatively high toxicity (56.7%, 76.7%, 66.7%, and 63.3%) against H. cunea at 100 µg/g of artificial diets. SDS-PAGE analysis revealed that a128kDa protein is associated with toxicity of Bt-2. Our result demonstrates a medium and strong larvicidal activity of Bt-2 against P. xylostella and H. cunea, respectively. Moreover, Bt-2 could be potentially produced using a cost-effective PBG medium which makes it an effective alternative biocontrol strategy to reduce chemical pesticide application.

Keywords: biocontrol, insect pests, larvae/nymph mortality, cost-effective media, aphis gossypii, plutella xylostella, hyphantria cunea, bacillus thuringiensi

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Authors: Sourav Ray, Christoph Stein, Marcus Weber

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A new class of drug candidates, initially derived from mathematical modeling of ligand-receptor interactions, activate the μ-opioid receptor (MOR) preferentially at acidic extracellular pH-levels, as present in injured tissues. This is of commercial interest because it may preclude the adverse effects of conventional MOR agonists like fentanyl, which include but are not limited to addiction, constipation, sedation, and apnea. Animal studies indicate the importance of taking the pH value of the chemical environment of MOR into account when designing new drugs. Hydrogen bonds (HBs) play a crucial role in stabilizing protein secondary structure and molecular interaction, such as ligand-protein interaction. These bonds may depend on the pH value of the chemical environment. For the MOR, antagonist naloxone and agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) form HBs with ionizable residue HIS 297 at physiological pH to modulate signaling. However, such interactions were markedly reduced at acidic pH. Although fentanyl-induced signaling is also diminished at acidic pH, HBs with HIS 297 residue are not observed at either acidic or physiological pH for this strong agonist of the MOR. Molecular dynamics (MD) simulations can provide greater insight into the interaction between the ligand of interest and the HIS 297 residue. Amino acid protonation states are adjusted to the model difference in system acidity. Unbiased and unrestrained MD simulations were performed, with the ligand in the proximity of the HIS 297 residue. Ligand-receptor complexes were embedded in 1-palmitoyl-2-oleoyl-sn glycero-3-phosphatidylcholine (POPC) bilayer to mimic the membrane environment. The occurrence of HBs between the different ligands and the HIS 297 residue of MOR at acidic and physiological pH values were tracked across the various simulation trajectories. No HB formation was observed between fentanyl and HIS 297 residue at either acidic or physiological pH. Naloxone formed some HBs with HIS 297 at pH 5, but no such HBs were noted at pH 7. Interestingly, DAMGO displayed an opposite yet more pronounced HB formation trend compared to naloxone. Whereas a marginal number of HBs could be observed at even pH 5, HBs with HIS 297 were more stable and widely present at pH 7. The HB formation plays no and marginal role in the interaction of fentanyl and naloxone, respectively, with the HIS 297 residue of MOR. However, HBs play a significant role in the DAMGO and HIS 297 interaction. Post DAMGO administration, these HBs might be crucial for the remediation of opioid tolerance and restoration of opioid sensitivity. Although experimental studies concur with our observations regarding the influence of HB formation on the fentanyl and DAMGO interaction with HIS 297, the same could not be conclusively stated for naloxone. Therefore, some other supplementary interactions might be responsible for the modulation of the MOR activity by naloxone binding at pH 7 but not at pH 5. Further elucidation of the mechanism of naloxone action on the MOR could assist in the formulation of cost-effective naloxone-based treatment of opioid overdose or opioid-induced side effects.

Keywords: effect of system acidity, hydrogen bond formation, opioid action, receptor activation

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Authors: L. Strauta, S. Muižniece-Brasava

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Even though legumes possess high nutritional value and have a rather high protein content for plant origin products, they are underutilized mostly due to their lengthy cooking time. To increase the presence of legume-based products in human diet, new extruded products were made of cow peas (Vigna unguiculata (L.) Walp.). But as it is known, adding different moisture content to flour before extrusion can change the physical properties of the extruded product. Experiments were carried out to estimate the optimal moisture content for cow pea extrusion. After extrusion, the pH level had dropped from 6.7 to 6.5 and the lowest hardness rate was observed in the samples with additional 9 g 100g^-1 of moisture - 28±4N, but the volume mass of the samples with additional 9 g100g^-1 of water was 263±3 g L^-1; all samples were approximately 7±1mm long.

Keywords: cow pea, extrusion–cooking, moisture, size

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Authors: Vilma Sasyte, Vilma Viliene, Asta Raceviciute-Stupeliene, Agila Dauksiene, Romas Gruzauskas, Virginijus Slausgalvis, Jamal Al-Saifi

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The aim of the present study was to investigate the effect of butyrate, acids, medium-chain fatty acids and plant extract mixture on performance, blood and gastrointestinal tract characteristics of laying hens’. For the period of 8 weeks, 24 Hisex Brown laying hens were randomly assigned to 2 dietary treatments: 1) control wheat-corn-soybean meal based diet (Control group), 2) control diet supplemented with the mixture of butyrate, acids, medium chain fatty acids and plant extract (Lumance®) at the level of 1.5 g/kg of feed (Experimental group). Hens were fed with a crumbled diet at 125 g per day. Housing and feeding conditions were the same for all groups and met the requirements of growth for laying hens of Hisex Brown strain. In the blood serum total protein, bilirubin, cholesterol, DTL- and MTL- cholesterol, triglycerides, glucose, GGT, GOT, GPT, alkaline phosphatase, alpha amylase, contents of c-reactive protein, uric acid, and lipase were analyzed. Development of intestines and internal organs (intestinal length, intestinal weight, the weight of glandular and muscular stomach, pancreas, heart, and liver) were determined. The concentration of short chain fatty acids in caecal content was measured using the method of HPLC. The results of the present study showed that 1.5 g/kg supplementation of feed additive affected egg production and feed conversion ratio for the production of 1 kg of egg mass. Dietary supplementation of analyzed additive in the diets increased the concentration of triglycerides, GOT, alkaline phosphatase and decreased uric acid content compared with the control group (P<0.05). No significant difference for others blood indices in comparison to the control was observed. The addition of feed additives in laying hens’ diets increased intestinal weight by 11% and liver weight by 14% compared with the control group (P<0.05). The short chain fatty acids (propionic, acetic and butyric acids) in the caecum of laying hens in experimental groups decreased compared with the control group. The supplementation of the mixture of butyrate, acids, medium-chain fatty acids and plant extract at the level of 1.5 g/kg in the laying hens’ diets had the effect on the performance, some gastrointestinal tract function and blood parameters of laying hens.

Keywords: acids, butyrate, laying hens, MCFA, performance, plant extract, psysiological state

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Authors: L. García, N. Cobas, M. López

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Acrylamide, a probable carcinogen, is formed in high-temperature processed food (>120ºC) when the free amino acid asparagine reacts with reducing sugars, mainly glucose and fructose. Cane juices' repeated heating would potentially form acrylamide during brown sugar production. This study aims to determine if using panela in yogurt cake preparation increases acrylamide formation. A secondary aim is to analyze the acrylamide concentration in four cake confections with different caloric sweetener ingredients: beet sugar (BS), cane sugar (CS), panela (P), and a panela and chocolate mix (PC). The doughs were obtained by combining ingredients in a planetary mixer. A model system made up of flour (25%), caloric sweeteners (25 %), eggs (23%), yogurt (15.7%), sunflower oil (9.4%), and brewer's yeast (2 %) was applied to BS, CS and P cakes. The ingredients of PC cakes varied: flour (21.5 %), panela chocolate (21.5 %), eggs (25.9 %), yogurt (18 %), sunflower oil (10.8 %), and brewer’s yeast (2.3 %). The preparations were baked for 45' at 180 ºC. Moisture was estimated by AOAC. Protein was determined by the Kjeldahl method. Ash percentage was calculated by weight loss after pyrolysis (≈ 600 °C). Fat content was measured using liquid-solid extraction in hydrolyzed raw ingredients and final confections. Carbohydrates were determined by difference and total sugars by the Luff-Schoorl method, based on the iodometric determination of copper ions. Finally, acrylamide content was determined by LC-MS by the isocratic system (phase A: 97.5 % water with 0.1% formic acid; phase B: 2.5 % methanol), using a standard internal procedure. Statistical analysis was performed using SPSS v.23. One-way variance analysis determined differences between acrylamide content and compositional analysis, with caloric sweeteners as fixed effect. Significance levels were determined by applying Duncan's t-test (p<0.05). P cakes showed a lower energy value than the other baked products; sugar content was similar to BS and CS, with 6.1 % mean crude protein. Acrylamide content in caloric sweeteners was similar to previously reported values. However, P and PC showed significantly higher concentrations, probably explained by the applied procedure. Acrylamide formation depends on both reducing sugars and asparagine concentration and availability. Beet sugar samples did not present acrylamide concentrations within the detection and quantification limit. However, the highest acrylamide content was measured in the BS. This may be due to the higher concentration of reducing sugars and asparagine in other raw ingredients. The cakes made with panela, cane sugar, or panela with chocolate did not differ in acrylamide content. The lack of asparagine measures constitutes a limitation. Cakes made with panela showed lower acrylamide formation than products elaborated with beet or cane sugar.

Keywords: beet sugar, cane sugar, panela, yogurt cake

Procedia PDF Downloads 66

Authors: Wirot Likittrakulwong, Pisit Poolprasert, Tossaporn Incharoen

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Germinated paddy rice (GPR) has the potential to be used as a feed ingredient. However, their properties have not been fully investigated. This paper examined the nutrient digestibility and the relationship to immune activity in Roman hens fed with GPR. It was found that true and apparent metabolizable energy (ME) values of GPR were 3.20 and 3.28 kcal/g air dry, respectively. GPR exhibited high content of phytonutrients, especially GABA. GPR showed similar protein profiles in comparison to non-germinated paddy rice. For immune activity, the feed with GPR enhanced the immune activity of Roman hens under high stocking density stress as evidenced by the activity of superoxide dismutase (SOD) and lysozyme activity. In this study, GPR is proved to be a good source of functional ingredient for chicken feed.

Keywords: germinated paddy rice, nutrient digestibility, immune activity, functional property

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Authors: Vanessa Funk, Steven Blum, Stephanie Cole, Jorge Maciel, Matthew Lux

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Paper-based synthetic gene circuits offer a new paradigm for programmable, fieldable biodetection. We demonstrate that by freeze-drying gene circuits with in vitro expression machinery, we can use complimentary RNA sequences to trigger colorimetric changes upon rehydration. We have successfully utilized both green fluorescent protein and luciferase-based reporters for easy visualization purposes in solution. Through several efforts, we are aiming to use this new platform technology to address a variety of needs in portable detection by demonstrating several more expression and reporter systems for detection functions on paper. In addition to RNA-based biodetection, we are exploring the use of various mechanisms that cells use to respond to environmental conditions to move towards all-hazards detection. Examples include explosives, heavy metals for water quality, and toxic chemicals.

Keywords: cell-free lysates, detection, gene circuits, in vitro

Procedia PDF Downloads 394

Authors: Mesfin Angaw Tesfay

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Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, Glycosylation

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Authors: M. Akoz, O. B. Citil, I. Aydin

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Colostrum secreted by the mammary glands after birth in the early days, a high proportion of fat, protein and ash containing a secretion containing low amounts of casein and lactose. Especially immunoglobulins contain high proportions. Maternal immunoglobulins own immune system to protect the newborn against neonatal disease until development are very important matter. However, colostrum is transferred to the offspring due to placental barrier in ruminants. Immunoglobulins are absorbed through the intestinal epithelium but absorption can vary under the influence of some factors. These factors are among the priority ones taking colostrum first time, amount, concentration, the metabolic status of the newborn. intestinal absorption of immunoglobulins occurs over the first 24 h high. Absorption from the gut after nine hours, 50% after 24 hours was only 11%. On the other hand pup's digestive system degrade the enzymes after 24 hours immunoglobulins. Bovine colostrum in the composition while basic immune IgG, IgA and IgM are also available. Total IgG in colostrum of ruminants, while in other species is a greater amount in blood serum.

Keywords: immunoglobulin, ruminants, colostrum, immune system

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Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson

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Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.

Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model

Procedia PDF Downloads 308

Authors: Nouha Bouayed Abdelmoula, Rim Louati, Nawel Abdellaoui, Balkiss Abdelmoula, Oldez Kaabi, Walid Smaoui, Samir Aloulou

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Background and Aims: Cardiofaciocutaneous syndrome (CFC) is an autosomal dominant disorder with the vast majority of cases arising by a new mutation of BRAF, MEK1, MEK2, or rarely, KRAS genes. Here, we report a rare Tunisian case of CFC syndrome for whom we identify SOS1 mutation. Methods: Genomic DNA was obtained from peripheral blood collected in an EDTA tube and extracted from leukocytes using the phenol/chloroform method according to standard protocols. High resolution melting (HRM) analysis for screening of mutations in the entire coding sequence of PTPN11 was conducted first. Then, HRM assays to look for hot spot mutations coding regions of the other genes of the RAS-MAPK pathway (RAt Sarcoma viral oncogene homolog Mitogen-Activated Protein Kinases Pathway): SOS1, SHOC2, KRAS, RAF1, KRAS, NRAS, CBL, BRAF, MEK1, MEK2, HRAS, and RIT1, were applied. Results: Heterozygous SOS1 point mutation clustered in exon 10, which encodes for the PH domain of SOS1, was identified: c.1655 G > A. The patient was a 9-year-old female born from a consanguineous couple. She exhibited pulmonic valvular stenosis as congenital heart disease. She had facial features and other malformations of Noonan syndrome, including macrocephaly, hypertelorism, ptosis, downslanting palpebral fissures, sparse eyebrows, a short and broad nose with upturned tip, low-set ears, high forehead commonly associated with bitemporal narrowing and prominent supraorbital ridges, short and/or webbed neck and short stature. However, the phenotype is also suggestive of CFC syndrome with the presence of more severe ectodermal abnormalities, including curly hair, keloid scars, hyperkeratotic skin, deep plantar creases, and delayed permanent dentition with agenesis of the right maxillary first molar. Moreover, the familial history of the patient revealed recurrent brain malignancies in the paternal family and epileptic disease in the maternal family. Conclusions: This case report of an overlapping RASopathy associated with SOS1 mutation and familial history of brain tumorigenesis is exceptional. The evidence suggests that RASopathies are truly cancer-prone syndromes, but the magnitude of the cancer risk and the types of cancer partially overlap.

Keywords: cardiofaciocutaneous syndrome, CFC, SOS1, brain cancer, germline mutation

Procedia PDF Downloads 153

Authors: M. Iqbal Hossain, Azharul Islam Khan, S. M. Rafiqul Islam, Tahmeed Ahmed

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Objective: Moderate acute malnutrition (MAM) is a major cause of morbidity and mortality in under-5 children of low-income countries. Approximately 14.6% of all under-5 mortality worldwide is attributed to MAM with >3 times increased risk of death compared to well-nourished peers. Prevalence of MAM among under-5 children in Bangladesh is ~12% (~1.7 million). Providing a diet containing adequate nutrients is the mainstay of treatment of children with MAM. It is now possible to process fish into fish peptides with longer shelf-life without refrigerator, known as ‘Fish Surimi peptide’ and this could be an attractive alternative to supply fish protein in the diet of children in low-income countries like Bangladesh. We conducted this study to assess the acceptability of Fish Surimi peptide given with various foods/meals in 2-5 years old children with MAM. Design/methods: Fish Surimi peptide is broken down from white fish meat using plant-derived enzyme and the ingredient is just fish meat consisted of 20 different kinds of amino acids including nine essential amino acids. In a convenience sample of 34 children we completed the study ward of Dhaka Hospital of icddr,b in Bangladesh during November 2014 through February 2015. For each child the study was for two consecutive days: i.e. direct observation of food intake of two lunches and two suppers. In a randomly and blinded manner and cross over design an individual child received Fish Surimi peptide (5g at lunch and 5g at supper) mixed meal [e.g. 30g rice and 30g dahl (thick lentil soup) or 60g of a vegetables-lentil-rice mixed local dish known as khichuri in one day and the same meal on other day without any Fish Surimi peptide. We observed the completeness and eagerness of eating and any possible side effect (e.g. allergy, vomiting, diarrhea etc.) over these two days. Results: The mean±SD age of the enrolled children was 38.4±9.4 months, weight 11.22±1.41 kg, height 91.0±6.3 cm, and WHZ was -2.13±0.76. Their mean±SD total feeding time (minutes) for lunch was 25.4±13.6 vs. 20.6±11.1 (p=0.130) and supper was 22.3±9.7 vs. 19.7±11.2 (p=0.297), and total amount (g) of food eaten in lunch and supper was found similar 116.1±7.0 vs. 117.7±8.0 (p=3.01) in A (Fish Surimi) and B group respectively. Score in Hedonic scale by mother on test of food given to children at lunch or supper was 3.9±0.2 vs. 4.0±0.2 (p=0.317) and on overall acceptance (including the texture, smell, and appearance) of food at lunch or supper was 3.9±0.2 vs. 4.0±0.2 (p=0.317) for A and B group respectively. No adverse event was observed in any food group during the study period. Conclusions: Fish Surimi peptide may be a cost effective supplementary food, which should be tested by appropriately designed randomized community level intervention trial both in wasted children and stunted children.

Keywords: protein-energy malnutrition, moderate acute malnutrition, weight-for-height z-score, mid upper arm circumference, acceptability, fish surimi peptide, under-5 children

Procedia PDF Downloads 412

Authors: Hategekimana Jean Paul, Irakoze Josiane, Ishimweyizerwe Valentin, Iradukunda Dieudonne, Uwanyirigira Jeannette

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Orange fleshed sweet potatoes (OFSP) are highly grown and are available plenty in rural and urban local markets and its contribution in reduction of food insecurity in Rwanda is considerable. But the postharvest loss of this commodity is a critical challenge due to its high perishability. Several research activities have been conducted on how fresh food commodities can be transformed into extended shelf life food products for prevention of post-harvest losses. However, such activity was not yet well studied in Rwanda. The aim of the present study was the processing of backed products from (OFSP)combined with wheat composite flour and assess the nutritional content and consumer acceptability of new developed products. The perishability of OFSP and their related lack during off season can be eradicated by producing cake, doughnut and bread with OFSP puree or flour. The processing for doughnut and bread were made by making OFSP puree and other ingredients then a dough was made followed by frying and baking while for cake OFSP was dried through solar dryer to have a flour together with wheat flour and other ingredients to make dough cake and baking. For each product, one control and three experimental samples, (three products in three different ratios (30,40 and50%) of OFSP and the remaining percentage of wheat flour) were prepared. All samples including the control were analyzed for the consumer acceptability (sensory attributes). Most preferred samples (One sample for each product with its control sample and for each OFSP variety) were analyzed for nutritional composition along with control sample. The Cake from Terimbere variety and Bread from Gihingumukungu supplemented with 50% OFSP flour or Puree respectively were most acceptable except Doughnut from Vita variety which was highly accepted at 50% of OFSP supplementation. The moisture, ash, protein, fat, fiber, Total carbohydrate, Vitamin C, reducing sugar and minerals (Sodium, Potassium and Phosphorus.) content was different among products. Cake was rich in fibers (14.71%), protein (6.590%), and vitamin c(19.988mg/100g) compared to other samples while bread found to be rich in reducing sugar with 12.71mg/100g compared to cake and doughnut. Also doughnut was found to be rich in fat content with 6.89% compared to other samples. For sensory analysis, doughnut was highly accepted in ratio of 60:40 compared to other products while cake was least accepted at ratio of 50:50. The Proximate composition and minerals content of all the OFSP products were significantly higher as compared to the control samples.

Keywords: post-harvest loss, OFSP products, wheat flour, sensory evaluation, proximate composition

Procedia PDF Downloads 62

Authors: V. Villa-Cruz, H. Pérez-Ladron de Guevara, J. E. Diaz-Díaz

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Static Light Scattering (SLS) was used as a method to analyse cow's milk raw, coming from the town of Lagos de Moreno, Jalisco, Mexico. This method is based on the analysis of the dispersion of light laser produced by a set of particles in solution. Based on the above, raw milk, which contains particles of fat globules, with a diameter of 2000 nm and particles of micelles of protein with 300 nm in diameter were analyzed. For this, dilutions of commercial milk were made (1.0%, 2.0% and 3.3%) to obtain a pattern of laser light scattering and also made measurements of raw cow's milk. Readings were taken in a sweep initial angle 10° to 170°, results were analyzed with the program OriginPro 7. The SLS method gives us an estimate of the percentage of fat content in milk samples. It can be concluded that the SLS method, is a quick method of analysis to detect adulteration in raw cow's milk.

Keywords: light scattering, milk analysis, adulteration in milk, micelles, OriginPro

Procedia PDF Downloads 375

Authors: Asim Cengiz, Dede Basturk, Hakan Ozalp

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Practicing recurring resistance workout such as may cause changes in human muscle. These changes may be because combination if several factors determining physical fitness. Thus, it is important to identify these changes. Several studies were reviewed to investigate these changes. As a result, the changes included positive modifications in amplified citrate synthase (CS) maximal activity, increased capacity for pyruvate oxidation, improvement on molecular signaling on human performance, amplified resting muscle glycogen and whole GLUT4 protein content, better health outcomes such as enhancement in cardiorespiratory fitness. Sprint training also have numerous long long-term changes inhuman body such as better enzyme action, changes in muscle fiber and oxidative ability. This is important because SV is the critical factor influencing maximal cardiac output and therefore oxygen delivery and maximal aerobic power.

Keywords: sprint, training, performance, exercise

Procedia PDF Downloads 303

Authors: Mohamed Fawzy Elzarei, Yousef Mohammed Al-Dakheel, Ali Mohamed Alseaf

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Modern molecular techniques, like single marker analysis for linked traits to these markers, can provide us with rapid and accurate genetic results. In the last two decades of the last century, the applications of molecular techniques were reached a faraway point in cattle, sheep, and pig. In goats, especially in our region, the application of molecular techniques is still far from other species. As reported by many researchers, microsatellites marker is one of the suitable markers for lie studies. The single marker linked to traits of interest is one technique allowed us to early select animals without the necessity for mapping the entire genome. Simplicity, applicability, and low cost of this technique gave this technique a wide range of applications in many areas of genetics and molecular biology. Also, this technique provides a useful approach for evaluating genetic differentiation, particularly in populations that are poorly known genetically. The expected breeding value (EBV) and yield deviation (YD) are considered as the most parameters used for studying the linkage between quantitative characteristics and molecular markers, since these values are raw data corrected for the non-genetic factors. A total of 17 microsatellites markers (from chromosomes 6, 14, 18, 20 and 23) were used in this study to search for areas that could be responsible for genetic variability for some milk traits and search of chromosomal regions that explain part of the phenotypic variance. Results of single-marker analyses were used to identify the linkage between microsatellite markers and variation in EBVs of these traits, Milk yield, Protein percentage, Fat percentage, Litter size and weight at birth, and litter size and weight at weaning. The estimates of the parameters from forward and backward solutions using stepwise regression procedure on milk yield trait, only two markers, OARCP9 and AGLA29, showed a highly significant effect (p≤0.01) in backward and forward solutions. The forward solution for different equations conducted that R2 of these equations were highly depending on only two partials regressions coefficient (βi,) for these markers. For the milk protein trait, four marker showed significant effect BMS2361, CSSM66 (p≤0.01), BMS2626, and OARCP9 (p≤0.05). By the other way, four markers (MCM147, BM1225, INRA006, andINRA133) showed highly significant effect (p≤0.01) in both backward and forward solutions in association with milk fat trait. For both litter size at birth and at weaning traits, only one marker (BM143(p≤0.01) and RJH1 (p≤0.05), respectively) showed a significant effect in backward and forward solutions. The estimates of the parameters from forward and backward solution using stepwise regression procedure on litter weight at birth (LWB) trait only one marker (MCM147) showed highly significant effect (p≤0.01) and two marker (ILSTS011, CSSM66) showed a significant effect (p≤0.05) in backward and forward solutions.

Keywords: microsatellites marker, estimated breeding value, stepwise regression, milk traits

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Authors: Tiziana Pepe, Gerardo Manfreda, Adriana Ianieri, Aniello Anastasio

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The sustainable exploitation of marine biological resources is among the most important objectives of the EU's Common Fisheries Policy (CFP). Currently, Europe imports about 65% of its fish products, indicating that domestic production does not meet consumer demand. Despite the availability of numerous commercially significant fish species, European consumption is concentrated on a limited number of products (e.g., sea bass, sea bream, shrimp). Many native species, present in large quantities in the Mediterranean Sea, are little known to consumers and are therefore considered ‘fishing by-products’. All the data presented so far indicate a significant waste of local resources and the overexploitation of a few fish stocks. It is therefore necessary to develop strategies that guide the market towards sustainable conversion. The objective of this work was to valorize underutilized fish species of the Mediterranean Sea through a multidisciplinary approach. To this end, three fish species were sampled: Atlantic Horse Mackerel (Trachurus trachurus), Bogue (Boops boops), and Common Dolphinfish (Coryphaena hippurus). Nutritional properties (water %, fats, proteins, ashes, salts), physical/chemical properties (TVB-N, histamine, pH), and rheological properties (color, texture, viscosity) were analyzed. The analyses were conducted on both fillets and processing by-products. Additionally, mitochondrial DNA (mtDNA) was extracted from the muscle of each species. The mtDNA was then sequenced using the Illumina NGS technique. The analysis of nutritional properties classified the fillets of the sampled species as lean or semi-fat, as they had a fat content of less than 3%, while the by-products showed a higher lipid content (2.7-5%). The protein percentage for all fillets was 22-23%, while for processing by-products, the protein concentration was 18-19% for all species. Rheological analyses showed an increase in viscosity in saline solution in all species, indicating their potential suitability for industrial processing. High-quality and quantity complete mtDNA was extracted from all analyzed species. The complete mitochondrial genome sequences were successfully obtained and annotated. The results of this study suggest that all analyzed species are suitable for both human consumption and feed production. The sequencing of the complete mtDNA and its availability in international databases will be useful for accurate phylogenetic analysis and proper species identification, even in prepared and processed products. Underutilized fish species represent an important economic resource. Encouraging their consumption could limit the phenomenon of overfishing, protecting marine biodiversity. Furthermore, the valorization of these species will increase national fish production, supporting the local economy, cultural, and gastronomic tradition, and optimizing the exploitation of Mediterranean resources in accordance with the CFP.

Keywords: mtDNA, nutritional analysis, sustainable fisheries, underutilized fish species

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Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

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Authors: Syed Sameer Aga, Saniya Nissar

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The Xeroderma pigmentosum group D gene (XPD) plays a key role in nucleotide excision repair (NER) pathway of the damaged DNA. Genetic polymorphisms in the coding region of the XPD gene may alter DNA repair capacity of the protein and hence can modulate the risk of colorectal cancer (CRC) risk. The aim of the study was to determine the genetic association of XPD Lys751Gln polymorphism with the risk of colorectal cancer (CRC) development. 120 CRC patients and 160 normal controls were assessed for genotype frequencies of XPD Lys751Gln polymorphism using PCR-RFLP technique. We observed a significant association (p < 0.05) between the XPD Lys751Gln polymorphism and the risk of developing CRC (p < 0.05). Additionally, Gln/Gln genotype of the XPD gene doubled the risk for the development of CRC [p < 0.05; OR=2.25 95% CI (1.07-4.7)]. Our results suggest that there is a significant association between the XPD Lys751Gln polymorphism and the risk of CRC.

Keywords: colorectal cancer, polymorphism, RFLP, DNA Repair, NER, XPD

Procedia PDF Downloads 216